REVIEW | doi:10.20944/preprints202310.0019.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: Punica granatum L.; pharmacokinetic; metabolic disorders; dyslipidaemia
Online: 1 October 2023 (08:42:27 CEST)
Modern research achievements support health-promoting effects of natural products and diets rich in polyphenols. Pomegranate (PG) (Punica granatum L.) contain a considerable number of bioactive compounds that exert a broad spectrum of beneficial biological activities, including antimicrobial, antidiabetic, antiobesity, and atheroprotective properties. In this view, the reviewed literature shows that PG intake might reduce insulin resistance, cytokine levels, redox gene expression, blood pressure elevation, vascular injuries and lipoprotein oxidative modifications. Lipid parameters corrective capabilities of PG-ellagitannins are also sufficiently reported to be significantly effective in reducing hyperlipidemia (TC, LDL-C, VLDL-C, and TAGs), plasma HDL-C concentrations, TC/HDL-C and LDL-C/HDL-C ratio. The health benevolent effects of pomegranate consumption appear to be produced through the amelioration of adipose tissue endocrine function, fatty acid utilization, GLUT receptor expression, paraoxonase activity enhancement, PPAR and NFκB modulation. Although the results from animal experiments are encouraging, human findings published in this field are inconsistent and still limited in many points of view. The present review discusses and provides a critical analysis of PG's bioeffects on metabolic syndrome components, type-2 diabetes, obesity, and dyslipidemia, as well as on some cardiovascular-related diseases. A brief overview of the pharmacokinetic properties, safety, and bioavailability of PG-ellagitannins is also included.
ARTICLE | doi:10.20944/preprints202310.0006.v1
Subject: Biology And Life Sciences, Plant Sciences Keywords: Abies balsamea; cold; conifer; fluorescence; galactosyldiacylglycerol; galactolipids; phospholipids; membrane injury; needle retention
Online: 1 October 2023 (08:11:54 CEST)
Needle abscission in balsam fir has been linked to both cold acclimation and changes in lipid composition. The overall objective of this research is to uncover lipid changes in balsam fir during cold acclimation and link those changes with postharvest abscission. Branches were collected monthly from September to December and assessed for cold tolerance via membrane leakage and chlorophyll fluorescence changes at -5, -15, -25, -35, and -45°C. Lipids were extracted and analyzed using mass spectrometry while postharvest needle abscission was determined gravimetrically. Cold tolerance and needle retention each significantly (P < 0.001) improved throughout autumn in balsam fir. There were concurrent increases in DGDG, PC, PG, PE, and PA throughout autumn and a decrease in MGDG. Those same lipids were strongly related to cold tolerance, though MGDG had the strongest relationship (R2 = 55.0% and 42.7% from membrane injury and chlorophyll fluorescence, respectively). There was a similar, albeit weaker, relationship between MGDG:DGDG and needle retention (R2 = 24.3%). Generally, a decrease in MGDG:DGDG ratio resulted in better cold tolerance and higher needle retention in balsam fir possibly due to increased membrane stability.
REVIEW | doi:10.20944/preprints202310.0012.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: pulmonary arterial hypertension; pulmonary arterial banding; right ventricle; preclinical studies
Online: 1 October 2023 (08:01:20 CEST)
Pulmonary hypertension (PH) is a progressive condition that affects pulmonary vessels, but its main prognostic factor is right ventricle (RV) function. Many mice/rat models are used for research in PAH, but results fail to translate in clinical trials. This study reviews studies that test interventions on pulmonary artery banding (PAB), a model of isolated RV disfunction, and PH models. Multiple tested drugs both improved pulmonary vascular hemodynamics in PH models and RV structure and function in PAB animals. PH models and PAB frequently exhibited similar results (73.1% concordance) with drugs other than endothelin receptor antagonists and phosphodiesterase inhibitors. RV systolic pressure accounted for most differences between PH models and PAB. On the other side, all results on RV fibrosis agreed (12 drugs). Macitentan, sildenafil and tadalafil improved most tested pathophysiological parameters in PH models, but almost none in PAB animals. Dapagliflozin was the only drug that improved no parameters. This review shows that many drugs currently under research for PAH have a cardioprotective effect on animals that may translate to humans. To improve the translational potential of drugs in this field, researchers should test them in multiple models, including PAB, while optimizing induction methods for human disease translation.
REVIEW | doi:10.20944/preprints202310.0005.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Mitochondria; immune response; activation; secretion; phagocytic
Online: 1 October 2023 (07:55:51 CEST)
Salmonella is the intracellular pathogen and etiological known for the cause of asymptomatic carriage, gastroenteritis, systemic disease (typhoid fever), in severe cases death may also occur. Altered function of mitochondria due to Salmonella infection impacts the immune response. Mitochondria which are recognized as the “powerhouse” of the cells are also know to play central role in the immune metabolism. Mitochondrial dynamics such as fusion, fission, signaling, transport and mitophagy influences the immune system and also maintains the cellular integrity. Mitochondrial DNA and Reactive oxygen species formation also elicits the inflammatory responses. This review discusses the immune metabolism and effect of altered mitochondrial dynamics in case of Salmonella infection.
ARTICLE | doi:10.20944/preprints202310.0003.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: microalgae; photoautotrophy; heterotrophy; growth; pigment content; photoacclimation; weak light effect
Online: 1 October 2023 (07:23:01 CEST)
Four strains of green microalgae (Scenedesmus acutus, Scenedesmus vacuolatus, Chlorella sorokiniana and Chlamydomonas reinhardtii) were compared for growth and pigment composition under photoautotrophic or heterotrophic conditions. Batch growth experiments were performed in multicultivators with online monitoring of optical density. For photoautotrophic growth, light-limited (CO2-sufficient) growth was analysed under different light intensities during exponential and deceleration growth phases. The specific growth rate, mesured during exponential phase, and the maximal biomass productivity, measured during deceleration phase, were not related to each other when different light intensities and different species were considered. This indicates species-dependent photoacclimation effects during cultivation time, which was confirmed by light-dependent changes in pigment contents and compositions when exponential and deceleration phases were compared. Except for C. reinhardtii, that does not grow on glucose, heterotrophic growth was promoted to similar extents by acetate or by glucose, however these two substrates led to different pigment compositions. Weak light increased pigment contents during heterotrophy in the four species, but was efficient in promoting growth only in S. acutus. C. sorokiniana and S. vacuolatus exhibited the best potential for heterotrophic biomass productivities both on glucose and acetate, with carotenoid (lutein) content being the highest in the former.
ARTICLE | doi:10.20944/preprints202309.1848.v1
Subject: Biology And Life Sciences, Aquatic Science Keywords: Carotenoid pigments; Fatty acids; Odontella aurita; Optimal growth condition; Ultrastructure; 18S rDNA
Online: 30 September 2023 (19:17:43 CEST)
Third-generation biomass production utilizing microalgae exhibits sustainable and environmentally friendly attributes, along with significant potential as a source of physiologically active compounds. However, the process of screening and localizing strains that are capable of producing high-value-added substances necessitates a significant amount of effort. In the present study, we have successfully isolated the indigenous marine diatom Odontella aurita OAOSH22 from the East coast of Korea. Subsequently, we conducted a comprehensive analysis of its morphological, molecular, and biochemical characteristics. Afterwards, comprehensive analysis was conducted on the morphological, molecular, and biochemical characteristics, along with testing for optimal growth conditions. The morphological characteristics of the isolate were observed using optical and electron microscopes, and it exhibited typical features of O. aurita. Additionally, the molecular phylogenetic inference derived from the sequence of the small-subunit 18S rDNA confirmed the classification of the microalgal strain as O. aurita. This isolate has been confirmed to contain 7.1 mg g-1 dry cell weight (DCW) of fucoxanthin, a powerful antioxidant substance, in terms of dry cell weight (DCW). In addition, this isolate contains 11.1 mg g-1 DCW of eicosapentaenoic acid (EPA), which is one of the nutritionally essential polyunsaturated fatty acids. Therefore, this indigenous isolate exhibits a significant potential as a valuable source of bioactive substances for various bio-industrial applications.
ARTICLE | doi:10.20944/preprints202309.2114.v1
Subject: Biology And Life Sciences, Aging Keywords: Human; lifespan; aging; consciousness; brain; integration
Online: 30 September 2023 (17:55:59 CEST)
Human longevity is an important but difficult goal due to the extreme complication of human body. If people could repeatedly transfer their consciousness from old bodies to new ones, their lifespan might be prolonged extremely. However, there are several difficulties which prevent the achievement of such a technique. In this paper, we propose an approach that can transfer human consciousness indirectly, and avoid or significantly relieve those difficulties. The strategy of our approach is to integrate the consciousnesses of two bodies (old and new) into a unique consciousness. By doing so, the consciousness is extended to the new body. After that, the consciousness shrinks and remains in the new body when the old body dies. This may also apply to integrating more than two bodies of human, or animal, or even across different species. After investigating thousands of literatures, we find quite strong evidence (related discoveries and technologies) that can support the proposed approach and its advantages. Beside prolonging human lifespan, the approach could also have other meaningful applications.
REVIEW | doi:10.20944/preprints202309.2115.v1
Subject: Biology And Life Sciences, Plant Sciences Keywords: Maize; Omics; Stresses; Resistance; Crop Improvement
Online: 30 September 2023 (10:26:37 CEST)
Maize (Zea mays L.) stands as one of the most extensively cultivated crops globally, offering significant contributions to food, animal feed, and biofuel production. Maize yield is significantly impacted by diverse climate and soil variables, yet its productivity faces hindrances due to abiotic stresses, including drought, salinity, extreme temperatures, and cold conditions. In confronting these hurdles, the field of crop breeding has undergone a transformation thanks to high-throughput sequencing technologies (HSTs). These advancements have streamlined the identification of beneficial quantitative trait loci (QTL), associations between markers and traits (MTAs), as well as genes and alleles that contribute to crop improvement. Presently, well-established omics techniques such as genomics, transcriptomics, proteomics, and metabolomics are being amalgamated into maize breeding studies. These approaches have unveiled new biological markers that are being employed to bolster maize's capacity to withstand an array of challenges. In this chapter, we explore the current knowledge regarding the morpho-physiological and molecular mechanisms that underlie the resistance and tolerance of maize to biotic and abiotic stresses. The focus is on utilizing omics techniques to augment maize's ability to withstand these challenges. Moreover, it emphasizes the significant potential of integrating various omics techniques to tackle the challenges presented by biotic and abiotic stress in maize productivity, contrasting with singular approaches.
REVIEW | doi:10.20944/preprints202309.1843.v2
Subject: Biology And Life Sciences, Anatomy And Physiology Keywords: disuse; fascicle; hypoplasia; longitudinal atrophy; muscle fibers; myofibril; myofilaments; radial atrophy; sarcomere
Online: 30 September 2023 (10:02:22 CEST)
The maintenance of skeletal muscle mass plays a fundamental role in health and issues associated with quality of life. Mechanical signals are one of the most potent regulators of muscle mass, with a decrease in mechanical loading leading to a decrease in muscle mass. This concept has been supported by a plethora of of human- and animal-based studies during the last 100 years and has resulted in the commonly used term of “disuse atrophy”. These same studies have also provided a great deal of insight into the structural adaptations that mediate disuse-induced atrophy. For instance, disuse results in radial atrophy of fascicles, and this is driven, at least in part, by radial atrophy of the muscle fibers. However, the ultrastructural adaptations that mediate these changes remain far from defined. Indeed, even the most basic questions, such as whether the radial atrophy of muscle fibers is driven by the radial atrophy of myofibrils and/or myofibril hypoplasia, have yet to be answered. In this review, we thoroughly summarize what is known about the macroscopic, microscopic, and ultrastructural adaptations that mediated disuse-induced atrophy and highlight some of the major gaps in knowledge that need to be filled.
ARTICLE | doi:10.20944/preprints202309.2166.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: L-asparaginase II; Bacillus subtilis; saline environment; biochemical characterization
Online: 30 September 2023 (08:39:19 CEST)
L-asparaginase from bacterial sources have been used in antineoplastic treatments and the food industry. A novel type II L-a sparaginase encoded by the N-truncated gene ansZP21 of halotolerant Bacillus subtilis CH11 isolated from Chilca salterns in Peru was expressed using a heterologous system in Escherichia coli BL21 (DE3)pLysS. The recombinant protein was purified using one-step nickel affinity chromatography and exhibited activity of 234.38 U mg-1 and a maximum catalytic activity at pH 9.0 and 60 °C. The enzyme shows a homotetrameric form with an estimated molecular weight of 155 kDa by gel filtration chromatography. The enzyme half-life at 60 °C was 3 h 48 min, and L-asparaginase retained 50% of initial activity for 24 h at 37 °C. The activity was considerably enhanced by KCl, CaCl2, MgCl2, mercaptoethanol, and DL-dithiothreitol (p-value < 0.01). Moreover, the Vmax and Km were 145.2 µmol mL-1 min-1 and 4.752 mM, respectively. These findings evidence a promising novel type II L-asparaginase for future industrial applications.
ARTICLE | doi:10.20944/preprints202309.2130.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: Artemisia; essential oil; GC-MS; AChE; BChE
Online: 30 September 2023 (08:17:03 CEST)
The essential oil (EO) of Artemisia plants contains a large number of bioactive compounds that are widely used. The aim of this study was to analyse the chemical composition of EOs of six Arte-misia plants collected in Croatia and to test their cholinesterase inhibitory potential. GC-MS analysis of EO of A. absinthium showed that the dominant compounds are cis-sabinyl acetate and cis-epoxy-ocimene; in EO of A. abrotanum it is borneol; in EO of A. annua it is artemisia ketone, camphor, and 1,8-cineole; in EO of A. arborescens it is camphor and chamazulene; in EO of A. verlotiorum it is cis-thujone, 1,8-cineole, and trans-thujone; in EO of A. vulgaris, it is trans-thujone and trans-epoxy-ocimene. EO of the five studied Artemisia species from Croatia is rich in mono-terpenoid compounds (1,8-cineole, artemisia ketone, cis-thujone, trans-thujone, cis-epoxy-ocimene, camphor, borneol, and cis-sabinyl acetate). EO of A. arborescens is also rich in chamazulene. The results also showed that the tested EOs have moderate cholinesterase inhibi-tion potential, especially the EOs of A. annua, A. vulgaris, and A. abrotanum. This is the first anal-ysis of the chemical composition of the EOs of four Artemisia plants and the first analysis of cho-linesterase potential for plants collected in Croatia.
ARTICLE | doi:10.20944/preprints202309.2168.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: brain ischemia; Alzheimer’s disease; CA3 area; amyloid protein precursor; amyloid; α-secretase; β-secretase; presenilin 1 and 2; tau protein; genes
Online: 30 September 2023 (08:01:45 CEST)
Understanding the phenomena underlying the non-selective susceptibility to ischemia of py-ramidal neurons in the CA3 area of the hippocampus is important from the point of view of elu-cidating the mechanisms of memory loss and the development of post-ischemic dementia. We used an ischemic model of Alzheimer's disease to study changes in amyloid protein precursor gene expression, its cleavage enzymes and tau protein in the CA3 area of the hippocampus af-ter a 10-minute brain ischemia with 12, 18, and 24-month survival. Quantitative reverse tran-scriptase PCR assay showed that the expression of all the genes that contribute to amyloid pro-duction was dysregulated within 2 years in the CA3 area of the hippocampus after ischemia. The expression of the amyloid protein precursor gene was above the control values at all times of the study. The expression of the α-secretase gene also exceeded the control values throughout the study. In contrast, the expression of the β-secretase gene reaching its maximum increase 12 months after ischemia, was below control values after 18 months and again above control values after 24 months of survival. Presenilin 1 and 2 gene expression was significantly elevated throughout the follow-up period, with peak expression of both genes occurring 12 months after ischemia. This suggests that the genes studied are involved in the non-amyloidogenic processing of amyloid protein precursor. Also, tau protein gene expression was significantly elevated throughout the observation period, and peak gene expression was present 12 months after is-chemia. Data indicate that an episode of brain ischemia with long-term survival causes damage and death of pyramidal neurons in the CA3 area of the hippocampus in a manner dependent on modified tau protein. Thus defining a new and important mechanism of pyramidal neuronal death in the CA3 area after ischemia. In addition tau protein gene modification after brain is-chemia is useful in identifying ischemic mechanisms occurring in Alzheimer's disease.
ARTICLE | doi:10.20944/preprints202309.2144.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: mitochondrial respiratory chain complexes; supercomplexes; cAMP/PKA signaling pathway; soluble adenylate cyclase; metabolic flux theory; mitochondrial membrane potential
Online: 30 September 2023 (08:01:36 CEST)
The current view of the mitochondrial respiratory chain complexes I, III and IV foresees the oc-currence of their assembly in supercomplexes providing additional functional properties as compared with randomly colliding isolated complexes. According to the plasticity model, these two structural states of the respiratory chain may interconvert influenced by the intracellular prevailing conditions and in previous studies we candidated the mitochondrial membrane po-tential as a factor controlling their dynamic balance. Here, we extended those studies investigating if and how the cAMP/PKA-mediated signalling influences the aggregation state of the respiratory complexes. Analysis of inhibitory-titration profiles of the endogenous oxygen consumption rates in intact HepG2 cells with specific inhibitors of the respiratory complexes was performed to quantify, in the framework of the metabolic flux theory, the corresponding control coefficients. The results attained inhibiting pharmacologically either PKA and sAC indicate that the reversible phosphorylation of the respiratory chain complexes/supercomplexes influences their assembly state in response to the membrane potential. This conclusion is supported by scrutiny of the available structure of the CI,CIII2,CIV respirasome enabling to map several PKA-target serine residues exposed to the matrix side of the complexes I, III and IV at the contact interfaces of the three complexes.
ARTICLE | doi:10.20944/preprints202309.0139.v4
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Immune escape; Vaccine efficacy; COVID-19; SARS-CoV-2
Online: 30 September 2023 (07:44:51 CEST)
(1) Background: We previously reported the development of a recombinant protein SARS-CoV-2 vaccine, consisting of the Receptor-Binding Domain (RBD) of the SARS-CoV-2 spike protein, adjuvanted with aluminum hydroxide (alum) and CpG oligonucleotides. In mice and non-human primates, our wild-type (WT) RBD vaccine induced high neutralizing antibody titers against the WT isolate of the virus, and, with partners in India and Indonesia it was later developed into two closely resembling human vaccines, Corbevax and Indovac. Here, we describe the development and characterization of a next-generation vaccine adapted to the recently emerging XBB variants of SARS-CoV-2. (2) Methods: We conducted preclinical studies in mice using a novel yeast-produced SARS-CoV-2 XBB.1.5 RBD subunit vaccine candidate formulated with alum and CpG. We examined the neutralization profile of sera obtained from mice vaccinated twice intramuscularly at a 21-day interval with the XBB.1.5-based RBD vaccine, against WT, Beta, Delta, BA.4, BQ.1.1, BA.2.75.2, XBB.1.16, XBB.1.5 and EG.5.1 SARS-CoV-2 pseudoviruses. (3) Results: The XBB.1.5 RBD/CpG/alum vaccine elicited a robust antibody response in mice. Furthermore, serum from vaccinated mice demonstrated potent neutralization against the XBB.1.5 pseudovirus as well as several other Omicron pseudoviruses. However, regardless of high antibody cross-reactivity by ELISA, the anti-XBB.1.5 RBD antigen serum showed low neutralizing titers against the WT and Delta virus variants. (4) Conclusions: Whereas we observed modest cross-neutralization against Omicron subvariants by sera from mice vaccinated with the WT RBD/CpG/Alum vaccine or with the BA.4/5-based vaccine, sera raised against the XBB.1.5 RBD showed robust cross-neutralization. These findings underscore the imminent opportunity for an updated vaccine formulation utilizing the XBB.1.5 RBD antigen.
ARTICLE | doi:10.20944/preprints202309.2142.v1
Subject: Biology And Life Sciences, Virology Keywords: phage therapy; phage inhibition; complement system; Pseudomonas aeruginosa; myophages
Online: 30 September 2023 (07:42:38 CEST)
Therapeutic phages are primarily chosen based on their in vitro bacteriolytic activity. Although anti-phage antibodies are known to inhibit phage infection, the influence of other immune system components is less well known. An important anti-bacterial and anti-viral innate immune system that may interact with phages is the complement system, a cascade of proteases that recognizes and targets invading microorganisms. In this study, we aimed to study the effects of serum components such as complement on the infectivity of different phages targeting Pseudomonas aeruginosa. We used a fluorescence-based assay to monitor the killing of P. aeruginosa by phages of different morphotypes in presence of human serum. Our results reveal that several myophages are inhibited by serum in a concentration-dependent way, while the activity of four podophages and one siphophage tested in this study is not affected by serum. By using specific nanobodies blocking different components of the complement cascade, we show that activation of the classical complement pathway is a driver of phage inhibition. To determine the mechanism of inhibition, we produced bioorthogonally labeled fluorescent phages to study their binding by means of microscopy and flow cytometry. We show that phage adsorption is hampered in presence of active complement. Our results indicate that interactions with complement may affect in vivo activity of therapeutically administered phages. A better understanding of this phenomenon is essential to optimize the design and application of therapeutic phage cocktails.
ARTICLE | doi:10.20944/preprints202309.2136.v1
Subject: Biology And Life Sciences, Parasitology Keywords: particle separation; parasite egg identification and quantification; diagnostic microfluidic device; extreme point-of-care
Online: 30 September 2023 (07:18:51 CEST)
A recently proposed single image parasite quantification (SIMPAQ) platform based on a Lab-On-a-Disc (LOD) device has been earlier successfully tested in field conditions demonstrating the efficiency in soil-transmitted helminths (STH) egg detection and analysis. A related study (Micromachines 2021, 12, 1032) revealed the effects that can limit the performance of a SIMPAQ method due to the action of the Euler and Coriolis forces, and the interaction of the moving eggs with the walls of the LOD chamber. Here we propose a new improved design that allows to overcome those limitations and enhance the yield of the SIMPAQ LOD device which is demonstrated in the experiments with synthetic particle model system and real parasite eggs.
ARTICLE | doi:10.20944/preprints202309.2152.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: MMP-9; MMP-9-1562C/T polymorphism; transcriptional regulation; brain; human neurons
Online: 30 September 2023 (07:16:03 CEST)
The MMP-9-1562C/T polymorphism influence incidence and course of many diseases of the central nervous system. We found, using luciferase assays and Q-RT-PCR technique, allele-specific in-fluence of MMP-9-1562C/T polymorphism on the MMP-9 promoter activity and on the expression of MMP-9 mRNA in human neurons derived from SH-SY5Y cells. Then, we have elucidated the mechanism responsible for the allele-specific action of the MMP-9-1562C/T polymorphism on transcriptional regulation of the MMP-9 gene using pull-down assay combined with mass spec-trometry analysis, EMSA and EMSA supershift techniques, as well as DsiRNA-dependent gene silencing. We have found that MMP-9 promoter activity and MMP-9 mRNA expression are reg-ulated in human neurons in the MMP-9-1562C/T allele-specific manner with stronger upregulation conferred by the C allele. Moreover, we have revealed that the allele-specific action of the MMP-9-1562C/T polymorphism on the neuronal MMP-9 expression is related to HDAC1 and ZNF384 transcriptional regulators. We show that HDAC1 and ZNF384 bind differentially to the C and the T alleles forming in vitro different regulatory complexes. Moreover, our data demonstrate that HDAC1 and ZNF384 downregulate differentially the MMP-9 gene promoter activity and MMP-9 mRNA expression in vivo in human neurons acting mostly via the T allele.
ARTICLE | doi:10.20944/preprints202309.2153.v1
Subject: Biology And Life Sciences, Plant Sciences Keywords: powdery mildew; wheat; GWAS; FarmCPU; MTAs; candidate genes
Online: 30 September 2023 (05:51:02 CEST)
Powdery mildew (PM), caused by the fungal pathogen Blumeria graminis f. sp. tritici (Bgt), significantly threatens global bread wheat production. Although the use of resistant cultivars is an effective strategy for managing PM, currently available wheat cultivars lack sufficient levels of resistance. To tackle this challenge, we conducted a comprehensive genome-wide association study (GWAS) using a diverse panel of 286 bread wheat genotypes. Over three consecutive years (2020-21, 2021-22, and 2022-23), these genotypes were extensively evaluated for PM severity under field conditions following inoculation with virulent Bgt isolates. The panel was previously genotyped using the Illumina 90K SNP Infinium iSelect SNP assay to obtain genome-wide SNP marker coverage. By applying FarmCPU, a multilocus mixed model, we identified a total of 113 MTAs located on chromosomes 1A, 1B, 2B, 3A, 3B, 4A, 4B, 5A, 5B, 6B, 7A, and 7B at a significance level of p≤0.001. Notably, four novel MTAs on chromosome 6B were consistently detected in 2020-21 and 2021-22 environments. Furthermore, within the confidence intervals of the identified SNPs, we identified 96 candidate genes belonging to different proteins including 12 disease resistance/host-pathogen interactions related protein families. Among these, protein kinases, leucine-rich repeats, and zinc finger proteins, were of particular interest due to their potential roles in PM resistance. These identified loci can serve as targets for breeding programs aimed at developing disease-resistant wheat cultivars.
ARTICLE | doi:10.20944/preprints202309.2134.v1
Online: 30 September 2023 (05:42:45 CEST)
Dipteryx spp. is an important species in reforestation in the Amazon. The objective of this study is to characterize and compare the relationships between dendrometric variables in Dipteryx spp. stands in the Western Amazon by fitting linear regression equations for total height and crown diameter. Six forest stands were evaluated in three municipalities. Dendrometric variables collected included diameter at 1.3 m height (dbh), total height (ht) and crown diameter (dc). Simple and multiple linear regression equations were fitted to characterize the relationships between ht and dc. The total aboveground biomass of Dipteryx spp. trees and the carbon stock of the stands were estimated. The general equations showed higher R² values, exceeding 0.7. The general equations for estimating ht and dc were significant for all coefficients. The trees averaged 22 t/ha of aboveground biomass in the stands. There was a variation in carbon sequestration potential among stands, ranging from 5.12 to 88.91 t CO2.ha-1. Single-input equations using dbh as an independent variable are recommended for estimating dc and ht for individual Dipteryx spp. stands. Stands in the Western Amazon play a significant role in carbon sequestration and accumulation. Trees can sequester an average of 4.8 tons of CO2 per year.
REVIEW | doi:10.20944/preprints202309.2138.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Hepatitis C virus; chronic HCV; antivirals; Sustained virological response; IFN therapy; Vaccines
Online: 30 September 2023 (05:39:36 CEST)
Hepatitis C virus (HCV) infection is a worldwide public health problem. Chronic infection by HCV can lead to liver cirrhosis or cancer. Although some immune-competent individuals can clear the virus, others develop chronic HCV disease due to viral mutations or an impaired immune response. IFNs type I and III and the signal transduction induced by them are essential for a proper antiviral effect. Research on the viral cycle and immune escape mechanisms have generated the basis of therapeutic strategies to achieve a sustained virological response (SVR). The first therapies were based on IFN-, then IFN-α plus ribavirin (IFN-RBV); then, pegylated-IFN--RBV (PEGIFNα-RIV) to improve cytokine pharmacokinetics. However, the maximum SVR was 60%, and several significant side effects were observed, decreasing the patients' treatment adherence. The development of direct-acting antivirals (DAAs) significantly enhanced SVR (> 90%); the compounds were able to inhibit HCV replication without significant side effects, even in pediatric populations. The management of coinfected patients HBV-HCV and HCV-HIV has also improved based on DAA and PEG-IFNα-RBV (HBV-HCV). CD4 cells are crucial for an effective antiviral response. IFNλ3, IL28B, TNF-α, IL-10, TLR-3, and TLR-9 gene polymorphisms are involved in viral clearance, therapeutic responses, and hepatic pathologies. Future research focuses on searching for strategies to circumvent resistance-associated substitution (RAS) to DAAs, develop new therapeutic schemes for different medical conditions, including organ transplant, and develop vaccines for long-lasting cellular and humoral responses with cross-protection to different HCV genotypes. The goal is to minimise the probability of HCV infection, HCV chronicity and hepatic carcinoma.
ARTICLE | doi:10.20944/preprints202309.2155.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: beraprost; feline; chronic kidney disease; aldosterone; NOS
Online: 30 September 2023 (05:18:01 CEST)
Objectives: The aim of the study was to evaluate the effects of oral administration of clinical dose of beraprost for feline CKD in healthy cats, and also to examine whether NOS inhibition reversed them. Methods: A placebo-controlled pharmacological sequential design study was carried out to assess plasma aldosterone and renin concentrations, mean blood pressure, heart rate, renal plasma flow, and renal vascular resistance. Results: Beraprost reduced plasma aldosterone when compared to the placebo (P < 0.05); this was reversed when NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) was added to beraprost treatment (P < 0.01). No differences in the plasma renin concentration or hemodynamic parameters were detected between beraprost and placebo. The correlation ratios (η2) showed opposite relationships between beraprost and added L-NAME effects on aldosterone concentration, mean blood pressure, heart rate, renal plasma flow, and renal vascular resistance (P < 0.001). Conclusions: In healthy cats, the clinical dose of beraprost suppresses plasma concentration of aldosterone, which can be reversed by inhibition of NOS.
ARTICLE | doi:10.20944/preprints202309.2097.v1
Subject: Biology And Life Sciences, Toxicology Keywords: cellular dynamics; multicellular agent-based model; computer simulation; developmental toxicity.
Online: 29 September 2023 (12:02:33 CEST)
Developmental hazard evaluation is an important part of assessing chemical risks during pregnancy. Toxicological outcomes from prenatal testing in pregnant animals result from complex chemical-biological interactions, and while New Approach Methods (NAMs) based on in vitro bioactivity profiles of human cells offer promising alternatives to animal testing, most of these assays lack cellular positional information, physical constraints, and regional organization of the intact embryo. Here, we engineered a fully computable model of the embryonic disc in the compucell3d.org modeling environment to simulate epithelial-mesenchymal transition of epiblast cells and self-organization of mesodermal domains (chordamesoderm, paraxial, lateral plate, posterior/extraembryonic). Cell fate in the model is determined by an autonomous homeobox (HOX) clock driven by morphogenetic signals (e.g., FGF, WNT, ATRA, CDX). Executing the model renders a quantitative cell-level computation of mesodermal subpopulations and consequences of perturbation based on known embryogeny. For example, synthetic perturbation of the control network rendered altered phenotypes (cybermorphs) mirroring experimental mouse embryology, with 50% reductions in FGF4, FGF8 and BMP4 signaling resulting in 86%, 98% and 59% reductions, respectively in the posterior mesodermal population, while ATRA exposure also resulted in a 78% decrease in this population. This model enables integration of in vitro chemical bioactivity data for specific molecular targets with known embryology to test mechanistic veracity and quantitative prediction of altered development.
ARTICLE | doi:10.20944/preprints202309.2095.v1
Subject: Biology And Life Sciences, Agricultural Science And Agronomy Keywords: AMF; BCF; PGPR; plant immune system; priming; root microbiome; RKNs
Online: 29 September 2023 (11:57:23 CEST)
Commercial formulates of beneficial microbes have been used to enrich the rhizosphere microbiome of tomato plants grown in pots located in a glasshouse. These plants have been subjected to attacks by soil-borne parasites, such as root-knot nematodes (RKNs), and herbivores, such as the miner insect Tuta absoluta. The development of both parasites and the symptoms of their parasitism were restricted in these plants with respect to plants left untreated. A mixture, named in the text as Myco, containing plant growth promoting rhizobacteria (PGPR), opportunistic biocontrol fungi (BCF), and arbuscular mycorrhizal fungi (AMF) was more effective in limiting pest damages than a formulate containing the sole AMF (Ozor). Therefore, Myco-treated plants inoculated with RKNs were taken as a model to further study which biocontrol agent (BCA) was specifically responsible of plant immunization and the molecular pathways exploited to achieve such an immunization. The PGPR contained in Myco were not able to reduce nematode infection, rather, they worsened symptoms on plants, compared with those observed on untreated plants. When plants were treated with suspensions of Bacillus subtilis at concentrations similar to those provided with Myco and then inoculated with RKNs, infection was actually more severe than that on untreated plants. Therefore, it was argued that both BCF and AMFwere the microorganisms that, colonizing roots, synergistically stimulate plant immune system against RKNs. Beneficial fungi lowered the activities of the defense supporting enzymes endochitinases and β-1,3-glucanase to be able to colonize the roots. However, as early as three days after nematode inoculation, these enzyme activities, and the expression of the encoding pathogenesis related genes (PR-2, PR-3) were found to be enhanced in roots with respect to not inoculated plants, thus indicating that plants had been primed against RKNs. Addition of paclobutrazol, which reduced salicylic acid (SA) levels in cells, and of diphenyliodonium chloride, which inhibits superoxides generation, completely abolished the repressive effect of Myco on nematode infection and presumably plant immune reaction. Inhibitors of copper enzymes and of the alternative cyanide-resistant respiration did not significantly alter resistance induction by Myco. When Myco-treated plants were subjected to a moderate water stress and inoculated with nematodes, they retained numbers of developed individuals in the roots similar to those present in regularly watered plants, in contrast to what occurred in roots of untreated stressed plants that hosted very few individuals because of poor nutrient availability.
REVIEW | doi:10.20944/preprints202309.2106.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: BBB; endothelial glycocalyx; enlarged perivascular spaces; metabolic syndrome; neuroinflammation; neurovascular unit; obesity; perivascular spaces; perivascular unit; type 2 diabetes mellitus
Online: 29 September 2023 (11:14:38 CEST)
Abstract: Astrocytes (ACs) are the most abundant human cell in the brain and importantly, are the master connecting and communicating cells that provide structural and functional support of brain cells at all levels of organization. Further, they are recognized to be the guardians and housekeepers of the brain. Protoplasmic perivascular astrocyte endfeet and their basal lamina form the delimiting outermost barrier (glia limitans) of the perivascular spaces in postcapillary venules and are important for the clearance of metabolic waste and comprise the glymphatic system, which is critically dependent on the proper waste removal by the pvACef polarized aquaporin 4 water channels. Also, the protoplasmic perisynaptic astrocyte endfeet (psACef) are important in cradling the neuronal synapses that serve to maintain homeostasis and serve a functional and supportive role for synaptic transmission. Enlarged perivascular spaces (EPVS) are emerging as important aberrant findings on magnetic resonance imaging (MRI) and associate with white matter hyperintensities, lacunes, aging, accepted to be biomarkers for cerebral small vessel disease, increased in obesity, metabolic syndrome, and type 2 diabetes. Knowledge is exponentially expanding regarding EPVS along with the glymphatic system, since EPVS are closely associated with impaired glymphatic function and waste removal from the brain to the cerebrospinal fluid and systemic circulation. This review intends to focus on how the pvACef play a crucial role in the development of EPVS.
ARTICLE | doi:10.20944/preprints202309.2104.v1
Subject: Biology And Life Sciences, Insect Science Keywords: extinct lauxaniid flies; age of Acalyptratae; Paleogene
Online: 29 September 2023 (10:57:05 CEST)
Hemilauxania parvula sp. nov., a new fossil species of the family Lauxaniidae (Diptera: Acalyptratae), is described and illustrated from Oise amber, France (Eocene, lower Ypresian, ca 53 Ma) and its relationship is discussed. Inasmuch as this first finding of a member of Schizophora in Oise amber probably represents the oldest known record of this group of Diptera, the age of Schizophora, based on the known fossil records, is discussed.
REVIEW | doi:10.20944/preprints202309.2093.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: Unmetabolized folic acid (UMFA); folic acid; one-carbon cycle; Methylation; epigenetic; carcinogenesis; aging; neurodevelopment; embryogenesis; folate
Online: 29 September 2023 (10:39:29 CEST)
This review delves into the intricate relationship between folate (Vitamin B9) intake, especially its synthetic form, folic acid, and its implications on health and disease. While folate plays a pivotal role in the one-carbon cycle, essential for DNA synthesis, repair, and methylation, concerns arise from its excessive intake. The literature underscores potential deleterious effects, such as an increased risk of carcinogenesis, disturbances in DNA methylation, and impacts on embryogenesis, pregnancy outcomes, neurodevelopment, and disease risk. Notably, these consequences stretch beyond the immediate effects, potentially influencing future generations through epigenetic reprogramming. We probe into the molecular mechanisms underlying these effects, including accumulation of unmetabolized folic acid, Vitamin B12 dependent mechanisms, altered one carbon metabolism, altered methylation patterns, and interactions with critical receptors and signaling pathways. Furthermore, we emphasize differences in the effects and mechanisms mediated by folic acid compared to natural folate. Given the widespread folic acid supplementation, it is imperative to further research its optimal intake levels, and the molecular pathways impacted by its excessive intake, ensuring the health and well-being of the global population.
ARTICLE | doi:10.20944/preprints202309.2066.v1
Subject: Biology And Life Sciences, Plant Sciences Keywords: Primula macrocalyx; Primula woronowii; Primula saguramica; Extraction; Stoichiometric Anticatalytic; Antioxidant; ABTS; DPPH; Neutrophils; Myeloperoxidase; Reactive oxygen species
Online: 29 September 2023 (10:28:36 CEST)
The goal of this article was to evaluate the antioxidant and anti-inflammatory like activities of three species from genus Primula L. growing in Georgia: Primula macrocalyx, Primula woronowii and Primula saguramica . First, fractions containing shoot and root parts were air-dried, grinded and extracted by 80% ethanol followed by condensation and freeze-drying. Second, further chromatographic separations were done on Diaion HP-20 with the following solvents: H2O, methanol (50% and 100 %) and ethylacetate 100% to obtain the expected fractions.
REVIEW | doi:10.20944/preprints202309.2089.v1
Subject: Biology And Life Sciences, Agricultural Science And Agronomy Keywords: maize; global nutrition; macronutrients; biofortification; genetic modification; maize processing
Online: 29 September 2023 (10:15:15 CEST)
This chapter explores the pivotal role of maize (Zea mays L.) in global nutrition, emphasizing its status as a staple food worldwide. It discusses maize's complex nutritional profile, including macronutrients, micronutrients, and dietary fiber. The chapter also covers biofortification efforts to enhance maize with essential nutrients, addresses nutritional concerns related to maize-centric diets, and explores genetic modification and agronomic strategies to boost nutrient-rich maize yields. Additionally, it delves into maize processing traditions, nutrient retention techniques, and the importance of promoting nutrient-rich maize varieties. The chapter outlines post-harvest strategies to combat nutrient loss and prevent contamination while emphasizing the significance of consumer education and diverse diets. Case studies illustrate the impact of improved maize nutrition and provide replicable strategies. The chapter concludes by considering emerging research and technology, identifying challenges, and proposing solutions to enhance maize's nutritional value in the future.
ARTICLE | doi:10.20944/preprints202309.2084.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: Collaborative cross (CC); Smad4 knock out; body weight gain; machine learning (ML); genotyping
Online: 29 September 2023 (08:44:37 CEST)
Background: Obesity and its attendant conditions have become major health problems worldwide and is currently ranked as the fifth most common leading cause of death globally. Complex environmental and genetic factors are the cause of the current obesity epidemic. Diet, lifestyle, chemical exposures, and other confounding factors are difficult to manage in humans. The mice model is helpful in researching genetics BW gain because genetic and environmental risk factors can be controlled in mice. Studies in mouse strains with various genetic backgrounds and established genetic structures provide unparalleled opportunities to find and analyze trait-related genomic loci. In this study, we used the Collaborative Cross (CC), a large panel of recombinant inbred mouse strains, to present a predictive study using CC mice SMAD4 knockout profiles to understand and effectively identify predisposition to body weight gain. Materials and Methods: Male C57Bl / 6J Smad4+/- mice were mated with female mice from 10 different CC lines to create F1 mice (Smad4+/- x CC). Body weight (BW) was measured weekly until week 16, then monthly until the end (week 48). The heritability H2 of the assessed traits was estimated and presented. Comparative analysis of various machine learning algorithms for predicting the BW changes and genotype of mice was conducted. Results and conclusions: Our data showed that the body weight records of F1 mice with different CC lines differed between wildtype and mutant Smad4 mice during the experiment. Genetic background affects weight gain and some lines gained more weight in the presence of Smad4 knockout, while others gained less, but in general, the mutation caused overweight mice, except for a few lines. In both control and mutant groups, female %BW had a higher heritability H2 value than males. Additionally, both sexes with wildtype genotypes showed higher heritability values than the mutant group. Logistic Regression provides the most accurate mouse genotype predictions using machine learning. We plan to validate the proposed method on more CC lines and mice per line to expand machine learning for BW prediction.
ARTICLE | doi:10.20944/preprints202309.2043.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Avipoxvirus; molecular characterization; phylogenetic analysis; viral introduction; Portugal
Online: 29 September 2023 (08:32:26 CEST)
Avipoxvirus (APV), a linear dsDNA virus belonging to the subfamily Chordopoxvirinae of the family Poxviridae, infects more than 278 species of domestic and wild birds. It is responsible for causing the avian pox disease, which is characterized by its cutaneous and diphtheric forms. With a high transmission capacity, it can cause high economic losses and damage to the ecosys-tem. Several diagnostic methods are available and vaccination of birds can be an effective pre-ventive measure. To update the molecular characterization and phylogenetic analysis of viruses isolated in Portugal between 2017 and 2023, ten APV-positive samples were analyzed. A P4b gene fragment was amplified by PCR and the nucleotide sequence of the amplicons was deter-mined by Sanger sequencing. The sequences obtained were aligned using ClustalW, and a max-imum likelihood phylogenetic tree was constructed. With this study, it was possible to verify that the analyzed sequences are distributed in subclades A1, A2, B1, and B3. Since some of them are quite similar to others from different countries and obtained in different years, it is possible to conclude that there have been several viral introductions in Portugal. Finally, it was possible to successfully update the data on avipoxviruses in Portugal.
ARTICLE | doi:10.20944/preprints202309.2037.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Nile tilapia; Oreochromis niloticus; Streptococcus agalactiae; formalin-killed vaccine; immunocompetence
Online: 29 September 2023 (08:29:48 CEST)
To investigate the early immune responses and explore the optimal vaccination periods, Nile tilapia at 1, 7, 14, 21, 28, 35, and 42 days after yolk sac collapse (DAYC), were immersed in formalin-killed Streptococcus agalactiae vaccine (FKV-SA). The results found that specific IgM by ELISA in 21 DAYC (0.108 g) was first detected at 336 h after vaccination (hav), whereas 28-42 DAYC (0.330 - 0.580 g), could be initially detected at 24 hav. qRT-PCR analysis of the TCRβ, CD4, MHCIIα, IgHM, IgHT, and IgHD genes at 21- 42 DAYC immunized with FKV-SA immersion route for 24, 168 and 336 hav, revealed that most immune-related genes were significantly higher in the vaccinated larvae in all DAYCs than the control larvae (P < 0.05) after 336 hav. Immunohistochemistry demonstrated the stronger signals of IgM in the gills, head kidney, and intestine tissues at 21, 28, and 35 DAYC were observed in all vaccinated larvae compared with the control. Interestingly, at all DAYCs, FKV-SA larvae exhibited significantly higher survival rates and an increased relative percent survival (RPS) than the control after challenge with viable S. agalactiae, particularly in larval fish that were immunized with FKV-SA for 168 and 336 hav (P < 0.05).
ARTICLE | doi:10.20944/preprints202309.2036.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: Blood brain barrier, Gliosis, Temporal lobe epilepsy, Inflammation, Epileptogenesis, Hippocampus, Endothelial cells, Pericytes, Basal membrane
Online: 29 September 2023 (08:26:25 CEST)
Temporal lobe epilepsy (TLE) is associated with reorganization of neuronal networks, gliosis, neuroinflammation, loss of integrity of the blood brain barrier (BBB) in the hippocampus in humans and animal models. More than 30% of epilepsies remain intractable and characterization of the molecular mechanisms involved in BBB dysfunction is essential to the identification of new therapeutic strategies. In this work, we induced status epilepticus in rats by injection of the proconvulsant drug pilocarpine that leads to TLE. Using RT-qPCR, double immunohistochemistry and confocal imaging, we studied at different time points of epileptogenesis (latent phase, 3, 7, 14 days; chronic phase, 1 and 3 months) the regulation of reactive glia and vascular markers. In the hippocampus, increased expression of mRNA encoding the glial proteins GFAP and Iba1 confirmed neuroinflammatory status. We report for the first time the concomitant induction in endothelial cells, pericytes and basal membrane of the BBB of specific proteins CD31, PDGFR and ColIV, that peaks at the same time points as inflammation. The altered expression of these proteins occurs early in TLE, during the latent phase, suggesting that they could be associated with early rupture and pathogenicity of the BBB that will contribute to the chronic phase of epilepsy.
HYPOTHESIS | doi:10.20944/preprints202309.2051.v1
Subject: Biology And Life Sciences, Ecology, Evolution, Behavior And Systematics Keywords: Lethal mutation; reproductively lethal mutation; reproductive bottlenecking; evolution; natural selection; phenotype; asexual reproduction; sexual reproduction; virus; bacterium; plant; insect, animal; eusociality
Online: 29 September 2023 (08:24:23 CEST)
Reproductively lethal mutations (RLMs) are mutations that, upon expression of the encoded lethal phenotypes, cause individuals carrying them to die or to be sterile. An underappreciated fact is that loss-of-function RLMs in protein-coding genes can be phenotypically shielded by favorable environments rendering these genes conditionally non-essential, or by their sister alleles in diploid organisms. Absent of rigorous mitigation, such phenotype-shielding causes the number of genes incurring RLMs to increase over time, and simultaneously allows each RLM to reach high allele frequencies in a conspecific population. Over-accumulation of RLMs then sets the population up for eventual concurrent expression of large numbers of RLMs, and massive deaths in rapid succession, possibly even population-level extinction. This hypothetical scenario in turn predicts that organismal lineages that evolved means to minimize the allele frequencies of phenotypically shielded RLMs are favored by natural selection. We argue that bottlenecking the genome copies destined for reproduction is a universal strategy adopted by all living beings to compel phenotype-based RLM purging. We further postulate that primitive RNA replicons must first evolve bottlenecked reproduction before evolving the capacity to encode diffusible products. In more complex, multicellular organisms, RLM management through bottlenecked reproduction gains additional reinforcement through sexual reproduction. In short, the evidence chronicled in this essay strongly suggest that the Bottleneck, Isolate, Amplify, Select (BIAS) principle, originally proposed to explain intracellular evolutionary dynamics of viruses, may be universally applicable to all living beings.
ARTICLE | doi:10.20944/preprints202309.2076.v1
Subject: Biology And Life Sciences, Food Science And Technology Keywords: FAME; SPME GC-MS; Compositional Analysis; Waterford Blaa; Sensory Analysis; Fatty Acid Profile; Volatile Profile
Online: 29 September 2023 (07:30:21 CEST)
Waterford Blaa is one of only seven Irish Protected Geographical Indication (PGI) products and the only Irish PGI bread. Currently, there are few published data characterising these products. This study aimed to determine whether any compositional or sensory markers could be used to differentiate between the three authorised producers of Waterford Blaa. Compositional testing was completed using AOAC standard methods; colour of top crust, bottom crust and crumb were identified with the Minolta Hunter lab colorimeter and texture profile analysis (TPA) was completed with an 80% compression test using the Instron 5544 to establish hardness, gumminess, and springiness. The fatty acid and aroma profiles were established in triplicate testing for the three authorised Blaa products using gas chromatography-flame ionisation detection and headspace-solid phase micro extraction-gas chromatography-mass spectrometry, respectively. As expected, results showed little variation in the overall composition of Waterford Blaa products. However, this study identified a unique fatty acid profile/volatile aroma profile for each Blaa. Specifically, the volatile profile showed potential differentiation between Blaa’s in terms of pentanal, furfural, 2-furanmethanol, 2-ethyl-1-hexanol and 2-pentyl furan. In addition, levels of hexanal and ethyl acetate were significantly different (p<0.05) in the crumb section of all three Blaa’s. This unique fingerprint has the potential to be used to identify fraudulent products claiming to be Waterford Blaa.
REVIEW | doi:10.20944/preprints202309.2069.v1
Subject: Biology And Life Sciences, Food Science And Technology Keywords: Postbiotics; prebiotics; probiotics; metabolites; lactic acid bacteria; fermentation
Online: 29 September 2023 (07:25:02 CEST)
Postbiotics are biologically active probiotic microorganisms that have produced non-viable bacterial products in the host. Eating a diet high in probiotic and prebiotic foods may help to ensure that the gut has an appropriate supply of these essential nutrients because postbiotics are produced when probiotics feed on prebiotics. Cell wall components and metabolites produced by living bacteria that have positive effects on the host are referred to as postbiotics. Postbiotics are functional bioactive chemicals that are produced in a matrix during anaerobic fermentation of organic nutrients like prebiotics. Postbiotics are low molecular weight soluble chemicals that are either produced after microbial cell lysis or secreted by live microflora as byproducts of this metabolic process. In general, the producer strains from bacterial and fungal species such as Lactobacillus, Bifidobacterium, Streptococcus, Akkermansia muciniphila, Saccharomyces boulardii, Eubacterium hallii, Faecalibacterium, etc. When handling and selling food items in an industrial setting, postbiotics can be used as functional components in foods, which has a number of advantages, including the potential to be added to some foods that are known to be harmful to probiotic survival. Postbiotic supplements have grown in popularity among customers because of their numerous health advantages, particularly food, cosmetic, and healthcare industries. Classification of postbiotics depends on various factors including type of microorganism, structural composition, and physiological functions. This review presents a succinct introduction, history, and concept of postbiotics, the salient features of postbiotics, classification, production, purification, characterization, biological functions, and applications of postbiotics in the food industry.
ARTICLE | doi:10.20944/preprints202309.2073.v1
Subject: Biology And Life Sciences, Endocrinology And Metabolism Keywords: horse; castration; steroid hormones; cartilage; bone; in vivo metabolism
Online: 29 September 2023 (05:25:34 CEST)
Castration of stallions is traditionally performed after puberty around the age of 2 years old. No studies have focused on the effects of early castration on osteoarticular metabolism. Thus, we sought to compare early castration (3 days after birth) with traditional castration (18 months of age) in horses. Testosterone and estradiol levels were monitored from birth to 33 months in these two groups. We quantified the levels of biomarkers of cartilage and bone anabolism (CPII and N-MID) and catabolism (CTX-I and CTX-II), of osteoarthritis (HA and COMP) and inflammation (IL-6 and PGE2). We revealed a lack of parallelism between testosterone and estradiol syntheses after birth and during puberty in both groups. An extra-gonadal synthesis of steroids was observed around the 28 month-mark, regardless of the castration age. We found the expression of estrogen receptor (ESR1) in cartilage and bone, whereas androgen receptor (AR) expression appeared to be restricted to bone. Nevertheless, with regards to osteoarticular metabolism, steroid hormone deprivation resulting from early castration showed no discernable impact on the levels of biomarkers related to bone and cartilage metabolism, nor on those associated with OA and inflammation. Consequently, our research demonstrated that early castration does not disrupt bone and cartilage homeostasis.
ARTICLE | doi:10.20944/preprints202309.2047.v1
Subject: Biology And Life Sciences, Anatomy And Physiology Keywords: anatomical section; magnetic resonance imaging; head; seabirds; Atlantic puffin
Online: 29 September 2023 (04:44:06 CEST)
The Atlantic puffin is a medium-sized seabird with black and white plumage and orange feet. It is distributed mainly along the northern Atlantic Ocean and due, among other reasons, to human activities, it is in a threatened situation and classified as a vulnerable species according to the International Union of Conservation of Nature (IUCN). In this study, we used a total of 20 carcasses of juvenile Atlantic puffins to perform anatomical cross sections and magnetic resonance images. Thus, an adequate description of the head was made, providing valuable information that could be helpful as a diagnostic tool for veterinary clinicians, who increasingly treat these birds in zoos, rehabilitation centers and even in the wild.
REVIEW | doi:10.20944/preprints202309.2030.v1
Subject: Biology And Life Sciences, Cell And Developmental Biology Keywords: Angiogenesis; blood vessels; endothelial cells; fetal cells; Flk1; migration; placenta; pregnancy; trophoblast; vasculogenesis; VEGF; VEGFR2
Online: 29 September 2023 (04:40:03 CEST)
Pregnancy entails bidirectional interactions between the developing fetus, the maternal tissues, and the organ systems. To this end, the phenomenon of migration of fetal cells (FCs) into the maternal circulation is poorly understood. Here, we review literature underlying the migration of FCs from the placenta to the maternal circulation, which is likely a dynamic process, including trophoblast invasion, placental angiogenesis, modulation of maternal immune responses, and enlargement of maternal organs. As placental neovascularization fosters direct connections between fetal and maternal circulatory systems, the trophoblast, a pivotal to placental development, adeptly deploys an array of invasive strategies to breach maternal tissue barriers, facilitating FC escapade into the maternal circulation. The intricate balance struck by the maternal immune system, which both acts as a guardian against potential foreign cell threats and orchestrates a niche conducive to FC survival and differentiation, is facilitated by finely tuned interactions among regulatory T cells, cytokines, and inhibitory receptors. FC presence in mothers’ circulation may be clinically relevant and unveil novel molecular participants like lncRNA, exosomes, and intricate signaling pathways that drive innovative clinical approaches for diagnostics and therapeutics. Ongoing research should reshape our knowledge of pregnancy and maternal-fetal health by improving our understanding of fetal-maternal interactions.
REVIEW | doi:10.20944/preprints202309.2000.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: Cluster of differentiation 74 (CD74); oncogenic fusion protein; fusion gene; CD74-ROS1; CD74-NTRK1; CD74-NRG1; CD74-NRG2a; CD74-PDGFRB; cancer.
Online: 29 September 2023 (04:35:12 CEST)
CD74 is a type II cell surface receptor found to be highly expressed in several hematological and solid cancers, due to its ability to activate pathways associated with tumor cell survival and proliferation. Over the past 16 years, CD74 is emerging as a commonly detected fusion partner in multiple oncogenic fusion proteins. Studies have found CD74 fusion proteins in a range of cancers including lung adenocarcinoma, inflammatory breast cancer, and pediatric acute lymphoblastic leukemia. To date, there are five known CD74 fusion proteins, CD74-ROS1, CD74-NTRK1, CD74-NRG1, CD74-NRG2a, and CD74-PDGFRB, with a total of 16 different variants, each with unique genetic signatures. Importantly, the occurrence of CD74 in the formation of fusion proteins has not been well explored despite the fact that ROS1 and NRG1 families utilize CD74 as the primary partner for the formation of oncogenic fusions. Fusion proteins known to be oncogenic drivers, including those of CD74, are typically detected, and targeted after standard chemotherapeutic plans fail and the disease relapses. The analysis reported herein provides insights into early intervention of CD74 fusions and highlight the need for improved routine assessment methods so that targeted therapies can be applied while they are most effective.
ARTICLE | doi:10.20944/preprints202309.2049.v1
Subject: Biology And Life Sciences, Parasitology Keywords: bats; digenea; Myotis spp.; Middle Volga region; Samarskaya Luka National Park
Online: 29 September 2023 (04:32:03 CEST)
Various bat species often occupy the same habitats. Cohabitation should induce different preferences in spatial or trophic components of the bat ecological niche, to reduce their competition. This determines the differences in the trematode fauna of Myotis spp. The purpose of our research was to study the biodiversity of trematodes in syntopic populations of five Myotis species in the Samarskaya Luka National Park. In 2005–2007 we studied 867 bat specimens by the methods of complete helminthological dissection. In total, 11 trematode species were found in Myotis bats. Only three trematode species, Plagiorchis koreanus, Prosthodendrium chilostomum and Parabascus duboisi were revealed in all studied Myotis spp. Prosthodendrium cryptolecithum was recorded for the first time in Russia. Digenean species diversity is higher in Myotis daubentonii and M. dasycneme. The trematode fauna of M. brandtii, M. nattereri and M. mystacinus is less diverse. The determining factor in the infection of bats by trematodes is feeding on semi-aquatic insects, possible second intermediate hosts of these parasites. Our results confirm the data that the ecological niches of the five Myotis species partially overlap. An analysis of the trematode fauna in Myotis spp. showed that in the Samarskaya Luka there may be weak competition for food items among bats.
ARTICLE | doi:10.20944/preprints202309.2013.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: Drosophila melanogaster; cocaine; methamphetamine; neurotransmitter concentration; locomotor sensitization; LC-MS/MS
Online: 29 September 2023 (04:30:31 CEST)
Due to similarities in genetics, cellular response, and behavior, Drosophila is used as a model organism in addiction research. A well described behavioral response examined in flies is the induced increase in locomotor activity after a single dose of volatilized cocaine (vCOC) and volatilized methamphetamine (vMETH) known as sensitivity, and the escalation of the locomotor response after the repeated dose, known as the locomotor sensitization. However, knowledge about how vCOC and vMETH affect different neurotransmitter systems over time is scarce. We used LC-MS/MS to systematically examine changes in the concentration of neurotransmitters, metabolites and non-metabolized COC and METH in the whole head homogenates of male flies one to seven hours after single and double vCOC or vMETH administrations. vMETH leads to complex changes in the levels of examined substances over time, while vCOC strongly and briefly increases concentrations of dopamine, tyramine and octopamine followed by a delayed degradation into N-acetyl dopamine and N-acetyl tyramine. The first exposure to psychostimulants leads to significant and dynamic changes in the concentrations relative to the second administration when they are more stable over several hours. Further investigations are needed to understand neurochemical and molecular changes post-psychostimulant administration.
ARTICLE | doi:10.20944/preprints202309.2058.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Caragana korshinskii Kom.; flavonoids; Taraxacum mongolicum Hand.-Mazz.; fermentation quality; antioxidant status; microbial community
Online: 29 September 2023 (04:26:21 CEST)
The present study was to investigate effects of total flavonoids from Taraxacum mongolicum Hand.-Mazz. (FT) on fermentation quality, antioxidant status and microbial community of Caragana korshinskii Kom. (CK) silage. CK was ensiled with no additive (CON), 1% FT and 2% FT on a fresh weight (FW) basis for 60 days. The results showed that 1% FT and 2% FT groups displayed higher DM content than CON group, and 2% FT group had the best effect on nutrient preservation. Compared with CON and 1% FT groups, 2% FT group exhibited the best silage fermentation quality and the highest antioxidant activity, including increased lactic acid, acetic acid concentrations and the activities of antioxidant enzymes, as well as decreased pH and the ammonia nitrogen (NH3-N) concentration. Moreover, 2% FT addition significantly affected the microbial community, such as increased abundance of Lactobacillus and decreased abundances of Pseudomonas and unidentified Cyanobacteria. The abundances of Lactobacillus parafarraginis and Lactobacillus brevis were negatively correlated to pH, while they were positively correlated with T-AOC, GSH-Px and CAT activities. In conclusion, 2% FT may be used as additives to inhibit the growth of undesirable microorganisms and promote fermentation quality and antioxidant activity of CK silage.
ARTICLE | doi:10.20944/preprints202309.1992.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: Cotton leaf curl Kokhran virus; microRNAs; RNA interference; prediction; computational algorithms; target binding sites; host-virus interaction
Online: 28 September 2023 (18:36:58 CEST)
Cotton leaf curl Kokhran virus (CLCuKoV) (genus, Begomovirus; family, Geminiviridae) is highly infectious, widespread and the most dangerous pathogen of cotton (Gossypium hirsutum L.) that is responsible to a serious disorder, cotton leaf curl disease (CLCuD). Begomoviruses are spread very efficiently by the whitefly Bemisia tabaci cryptic species, causing economic losses to cotton crop, all over the world. The ‘Lucknow’ strain of CLCuKoV has emerged as a divergent isolate that could cause CLCuD. The monopartite ssDNA genome of CLCuKoV-Lu (2.7 Kb) contains six open reading frames (ORFs) that was shown to encode four major proteins. RNA interference (RNAi)-based antiviral innate immunity is a sequence-specific biological phenomenon and a powerful tool to control plant viruses. The present study aims to determine cotton locus-derived microRNAs (ghr-miRNAs) that are identified for targeting the CLCuKoV-Lu ss-DNA-encoded mRNAs using a predictive approach that involves four computational algorithms, miRanda, RNA22, psRNATarget and RNAhybrid. Mature ghr-miRNA sequences (n=80) from allotetraploid upland cotton (2n = 4x = 52) were selected from the miRBase and were tested for alignment with the CLCuKoV-Lu genome. Among the 80 cotton locus-derived ghr-miRNAs evaluated, only one consensus cotton locus-derived ghr-miRNA (ghr-miR2950) was concluded to have effective ghr-miRNA target site at common nucleotide position 82 in the CLCuKoV-Lu genome respectively, using a stringent criterion, identified by all the algorithms used. The miRNA targeting is reliant on base pairing of miRNA-mRNA target pairings. Conservation of the hybridization binding site of the predicted ghr-miR2950 was validated using multiple sequence alignment within all the strains of CLCuKoV. We constructed a regulatory interaction network of miRNA–mRNA to identify novel targets. The efficacy of the predicted miRNAs against CLCuKoV-Lu was evaluated by RNAi-mediated targeted mRNA cleavage. The current investigated miRNA targets provide evidence for the development of CLCuD-resistant cotton plants.
ARTICLE | doi:10.20944/preprints202309.1976.v1
Subject: Biology And Life Sciences, Horticulture Keywords: pineapple; ripening; yellowing; fruit quality; metabolomics; transcriptomics
Online: 28 September 2023 (18:24:50 CEST)
To understand the molecular mechanism underlying yellowing in pineapples during ripening, coupled with alterations in fruit quality, comprehensive metabolome and transcriptome investigations were carried out. These investigations were conducted using the pulp samples collected at three distinct stages of maturity: young fruit (YF), mature fruit (MF), and fully mature fruit (FMF). This study revealed a noteworthy increase in the levels of total phenols and flavones, coupled with a concurrent decline in lignin and total acid content, as the fruit transitioned from YF to FMF. Furthermore, the analysis yielded 167 differentially accumulated metabolites (DAMs) and 2194 differentially expressed genes (DEGs). Kyoto Encyclopedia of Genes and Genomes enrichment analysis based on DAMs and DEGs revealed that the biosynthesis of plant secondary metabolites, particularly the flavonol, flavonoid, and phenypropanoid pathways, plays a pivotal role in fruit yellowing. Additionally, a comprehensive regulatory network encompassing genes that contribute to metabolisms of flavones, flavonols, lignin, and organic acids was proposed. This network sheds light on the intricate processes that underlies fruit yellowing and quality alterations. These findings enhance our understanding of the regulatory pathways governing pineapple ripening and offer valuable scientific insight into the molecular breeding of pineapples.
ARTICLE | doi:10.20944/preprints202309.1919.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: influenza; vaccines; antibodies; neuraminidase-inhibition; persistence.
Online: 28 September 2023 (17:59:45 CEST)
Neuraminidase (NA)-based immunity could reduce the harmful impact of novel antigenic variants of influenza viruses. Detection of neuraminidase-inhibiting (NI) antibodies in parallel with anti-hemagglutinin (HA) antibodies may enhance research of immunogenicity and duration of antibody response to influenza vaccines. To assess anti-NA antibodies after vaccination with seasonal inactivated influenza vaccines, we used the enzyme-linked lectin assay, and anti-HA antibodies were detected in hemagglutination inhibition assay. The dynamics of anti-NA antibody response differed depending on the virus subtype: antibodies to A/H3N2 virus neuraminidase increased later than antibodies to A/H1N1pdm09 subtype neuraminidase, and persisted longer. In contrast to HA antibodies, the fold increase in antibody titers to NA after vaccination poorly depended on the preexisting level. At the same time, NA antibody level after vaccination directly correlated with titers before vaccination. Difference was found in response to NA antigen between split and subunit adjuvanted vaccines and in NA functional activity in the vaccine formulations.
ARTICLE | doi:10.20944/preprints202309.1941.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: DNA repair; genetic polymorphism; head and neck cancer; radiotherapy; survival; CHEK1; MRE11; XRCC5; XRCC6; RAD51; LIG4; ATM; TP53; NBS1
Online: 28 September 2023 (17:54:16 CEST)
Radiotherapy and cisplatin-based chemotherapy belong to the main treatment modalities for head and neck squamous cell carcinoma (HNSCC), and induce cancer cell death by generating DNA damage, including the most severe double strand breaks (DSBs). Alterations in DSB response and repair genes may affect individual DNA repair capacity and treatment sensitivity, contributing to therapy resistance and poor prognosis often observed in HNSCC. In this study, we investigated the association of a panel of single nucleotide polymorphisms (SNPs) in 20 DSB signaling and repair genes with therapy results and prognosis in 505 HNSCC patients treated non-surgically with DNA damage-inducing therapies. In the multivariate analysis, there were a total of 14 variants associated with overall, locoregional recurrence-free or metastasis-free survival. Moreover, we identified 10 of these SNPs as independent predictors of therapy failure and unfavorable prognosis in the whole group or in two treatment subgroups. They were MRE11A rs2155209, XRCC5 rs828907, RAD51 rs1801321, rs12593359, LIG4 rs1805388, CHEK1 rs558351, TP53 rs1042522, ATM rs1801516, XRCC6 rs2267437 and NBS1 rs2735383. Only CHEK1 rs558351 remained statistically significant after correction for multiple testing. These results suggest that specific germline variants related to DSB response and repair may be potential genetic modifiers of therapy effects and disease progression in HNSCC treated with radiotherapy and cisplatin-based chemoradiation.
ARTICLE | doi:10.20944/preprints202309.1907.v1
Subject: Biology And Life Sciences, Food Science And Technology Keywords: Bisphenol A diglycidyl ether; derivatives; broad-spectrum polyclonal antibodies; AuNPs; immunochromatographic; canned food
Online: 28 September 2023 (17:24:06 CEST)
Bisphenol A diglycidyl ether (BADGE) is widely existed in the inner coating of canned foods. It migrates into food and generate various derivatives in the process of storage, such as Bisphenol A (2, 3-dihydroxypropyl) glycidyl ether (BADGE·H2O), Bisphenol A (3-chloro-2-hydroxypropyl) glycidyl ether (BADGE·HCl) and Bisphenol A (3-chloro-2-hydroxypropyl) (2, 3-dihydroxypropyl) glycidyl ether (BADGE·HCl·H2O), which have negative effects on human health. A gold nanoparticle-based immu-nochromatographic assay for simultaneous detection of BADGE and its derivatives was developed by using a broad-spectrum polyclonal antibody, and the detection can be finished in 15 min. The visuali-zation of results was processed by Adobe Photoshop CC software to achieve quantitative analysis and the detection limit (IC15) is 0.97ng/mL. The recoveries of BADGE and its derivatives at various spiking levels in canned food samples ranged from 79.86% to 93.81%. The detection results of the proposed immu-nochromatographic assay were also validated by HPLC analysis, and got good consistency (R2=0.9580).
ARTICLE | doi:10.20944/preprints202309.2010.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: Beeswax alcohol (BWA); High-density lipoproteins (HDL); Reconstituted HDL; Carboxymethyllysine (CML); Zebrafish; Embryo
Online: 28 September 2023 (13:15:56 CEST)
The antioxidant and anti-inflammatory abilities of beeswax alcohol (BWA) are well reported in animal and human clinical studies, with a significant decrease in malondialdehyde (MDA) in blood, reduction of liver steatosis, and decrease in insulin. On the other hand, there is insufficient information to explain the in vitro antioxidant activity of BWA because it is extremely insoluble in aqueous buffer system. The BWA mixture was incorporated into reconstituted HDL (rHDL) with apoA-I, and the physiological functions of BWA in a water system were evaluated. After synthesis of rHDL with palmitoyloleoyl phosphatidylcholine (POPC), cholesterol, apolipoprotein A-I (apoA-I), and the BWA at molar ratios of 95:5:1:0 (rHDL-0), 95:5:1:0.5 (rHDL-0.5), and 95:5:1:1 (rHDL-1) for POPC:FC:apoA-I:BWA, the particle size of rHDL-1 was increased 15% compared to rHDL-0. rHDL-1 exhibited the strongest anti-glycation activity, up to 18% less glycation than rHDL-0 treated HDL3, with the highest protection of apoA-I from proteolytic degradation: a 28% larger band intensity than that of rHDL-0 treated HDL3 in the presence of fructose (final 250 mM). The antioxidant ability to inhibit cupric ion-mediated LDL oxidation increased as the BWA content in rHDL increased. The rHDL-1-treated LDL exhibited the smallest oxidation extent in electromobility with the least quantification of oxidized species (MDA). The antioxidant activities of HDL, ferric ion reduction ability (FRA), and paraoxonase (PON) were enhanced by the BWA in rHDL treatment with a concomitant increase in the molar ratio. rHDL-1-treated HDL showed 20–22% higher FRA and PON activities than rHDL-0-treated HDL. A microinjection of each rHDL into zebrafish embryos was performed in the presence of carboxymethyllysine (CML). The rHDL-1 injected embryo exhibited the highest survivability (~63%), whereas the CML alone group showed 28% survivability. A higher BWA content in rHDL helped neutralize the CML toxicity, resulting in higher survivability and normal developmental morphology. In contrast, the CML alone injected embryo showed severe retardation of the developmental speed and morphological defect. The CML-alone group showed the highest extent of reactive oxygen species (ROS) production and cellular apoptosis in embryos, but a co-injection of rHDL-1 resulted in a remarkable decrease in ROS and apoptosis. The dermal application of rHDL containing BWA resulted in higher potent wound-healing activity in a dose-dependent manner with decreased reactive oxygen species and cellular apoptosis in the cutaneous wound area in the presence of CML. Conclusively, incorporating BWA in rHDL significantly enhanced the anti-glycation and antioxidant activities in rHDL via more stabilization of apoA-I with a larger particle size. The rHDL containing BWA facilitated enforced inherent antioxidant ability of HDL and anti-inflammatory activity to suppress the CML toxicities in zebrafish embryos and to ameliorate CML-aggravated chronic wounds in adult zebrafish.
REVIEW | doi:10.20944/preprints202309.2005.v1
Subject: Biology And Life Sciences, Aquatic Science Keywords: Risk assessment; tidal stream energy; environmental effects; collision risk; marine renewable energy
Online: 28 September 2023 (13:04:49 CEST)
Commercial development of tidal stream energy is hampered by technical and financial challenges, and impeded by uncertainty about potential environmental effects that drive environmental risk assessments and permitting (consenting) processes. The effect of greatest concern for operational tidal stream energy devices is the potential for marine animals to collide with turbine blades, resulting in injury or death. Due to the turbulent and often turbid waters that frequently characterize tidal turbine sites, there is an absence of empirical evidence about collisions with marine animals. This paucity of observations often leads to risk-averse permitting decisions that further restrict the deployment of tidal energy devices that are needed to collect this evidence. This paper relies on the framework of stressors and receptors that is used widely in marine energy studies and outlines a stepwise probabilistic methodology that applies existing knowledge to further elucidate the risk to marine animals from operational tidal turbines. A case study using striped bass from the Bay of Fundy, Canada, accompanies the methodology, to partially demonstrate its application.
ARTICLE | doi:10.20944/preprints202309.1922.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: polyhydroxyalkanoate; PHBHHx; CO2; Cupriavidus necator; hydrogen-oxidizing bacterium; high cell density cultivation; jar-cultivation
Online: 28 September 2023 (13:03:22 CEST)
The copolyester of 3-hydroxybutyrate (3HB) and 3-hydoxyhexanoate (3HHx), PHBHHx, is a bi-odegradable plastic that has a high flexibility, softness, wide process window and marine bio-degradability. PHBHHx is usually produced from structurally related carbon sources such as vegetable oils or fatty acids but not from inexpensive carbon sources like sugars. In the previous studies, we demonstrated that engineered strains of hydrogen-oxidizing bacterium, Cupriavidus necator synthesized PHBHHx with high cellular content from not only sugars but also from CO2 as the sole carbon source in the flask culture. In this study, highly efficient production of PHBHHx from CO2 was investigated by pH-stat jar cultivation of the recombinant C. necator strains with feeding the substrate gas mixture (H2/O2/CO2=8:1:1) to complete mineral medium in recycled-gas closed-circuit culture system. As the result with the strain MF01/pBPP-ccrMeJAc-emd, the dry cell mass and PHBHHx concentration reached up to 59.62 ± 3.18 g･L-1 and 49.31 ± 3.14 g･L-1, respectively after 212 h of jar cultivation with limited addition of ammonia and phosphate solutions. The 3HHx composition was close to 10 mol% which is suitable for practical applica-tions. It is expected that autotrophic cultivation of the recombinant C. necator can be a feasible process for mass production of PHBHHx from CO2.
ARTICLE | doi:10.20944/preprints202309.2006.v1
Subject: Biology And Life Sciences, Cell And Developmental Biology Keywords: equine; endothelial; angiogenesis; comparative
Online: 28 September 2023 (13:01:45 CEST)
Understanding the factors which control endothelial cell (EC) function and angiogenesis in the horse is crucial for developing the horse as a disease model but equine ECs remain poorly studied. In this study we have optimised methods for the isolation and culture of equine aortic endothelial cells (EAoECs) and characterised their angiogenic functions in vitro. Mechanical dissociation, followed by magnetic purification using an anti-VE-cadherin antibody, resulted in EC-enriched cultures suitable for further study. Fibroblast growth factor 2 (FGF2) increased EAoEC proliferation rate and stimulated scratch wound closure and tube formation by EAoECs on extracellular matrix. Pharmacological inhibitors of FGFR1 (SU5402) or MEK (PD184352) blocked FGF2-induced ERK1/2 phosphorylation and functional responses, suggesting that these are dependent on FGFR1/MEK-ERK signalling. In marked contrast, VEGF-A had no effect on EAoEC proliferation, migration or tubulogenesis and did not promote ERK1/2 phosphorylation, indicating a lack of sensitivity to this classical pro-angiogenic growth factor. Gene expression analysis showed that, unlike human ECs, FGFR1 is expressed by EAoECs at a much higher level than both VEGFR1 and VEGFR2. These results suggest a predominant role for FGF2 versus VEGF-A in controlling the angiogenic functions of equine ECs. Collectively, our novel data provide a sound basis for studying angiogenic processes in the horse and lay the foundations for comparative studies of EC biology in horses versus humans.
ARTICLE | doi:10.20944/preprints202309.2002.v1
Subject: Biology And Life Sciences, Agricultural Science And Agronomy Keywords: fusarium wilt; bacillus; quercetin; antagonism; induced resistance
Online: 28 September 2023 (11:48:48 CEST)
Fusarium wilt diseases severely influence the growth and productivity of numerous crop plants. The consortium of antagonistic rhizospheric Bacillus strains and quercetin were evaluated imperatively as a possible remedy to effectively manage the Fusarium wilt disease of tomato plants. The selection of Bacillus strains was made based on in-vitro antagonistic bioassays against Fusarium oxysporum f.sp. lycoprsici (FOL). Whereas, quercetin was selected after screening a library of phytochemicals during in-silico molecular docking analysis using tomato LysM receptor Kinases “SILKY12” based on its dual role in symbiosis and plant defense responses. After the selection of test materials, pot trials were conducted where tomato plants were provided consortium of Bacillus strains as soil drenching and quercetin as a foliar spray in different concentrations. The combined application of consortium () and quercetin (1.0 mM) reduced the Fusarium wilt disease index up to 69% also resulting in increased plant growth attributes. Likewise, the imperative application of the Bacillus consortium and quercetin (1.0 mM) significantly increased total phenolic contents and activities of the enzymes of the phenylpropanoid pathway. Non-targeted metabolomics analysis was performed to investigate the perturbation in metabolites. FOL pathogen negatively affected a range of metabolites including carbohydrates, amino acids, phenylpropanoids, and organic acids. Thereinto, combined treatment of Bacillus consortium and quercetin (1.0 mM) ameliorated the production of different metabolites in tomato plants. These findings prove the imperative use of Bacillus consortium and quercetin as an effective and sustainable remedy to manage Fusarium wilt disease of tomato plants and to promote the growth of tomato plants under pathogen stress conditions.
ARTICLE | doi:10.20944/preprints202309.1964.v1
Subject: Biology And Life Sciences, Virology Keywords: Biothermodynamics; Virus-host interaction; COVID-19; Pandemic; Variant under monitoring; Pathogenicity; Pathogen; August 2023
Online: 28 September 2023 (11:23:34 CEST)
Similarly to a phoenix, SARS-CoV-2 has appeared periodically in waves. The new variants that appeared through mutations have during the 4 years of the pandemic suppressed earlier variants, causing new waves of the pandemic. The Omicron BA.2.86 Pirola variant is the latest in the sequence of SARS-CoV-2 variants, which appeared in 2023. The BA.2.86 variant has started to spread rapidly and we are witnesses of a new epidemic wave. In this short period, an increased infectivity was noticed, which results in rapid spreading and decreased pathogenicity, which results in a lower number of severe cases. However, in the public there is a fear of further development of the epidemic. This analysis was made with the goal to assess the risks in the period of late 2023. Mutations that were developed by the BA.2.86 variant have led to a change in empirical formula and thermodynamic properties. It seems that there is no ground for fear of an extensive spreading of severe forms, but there are reasons for caution and monitoring of the spreading of the epidemic and potential appearance of new mutations.
ARTICLE | doi:10.20944/preprints202309.1988.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Clostridium perfringens; necrotic enteritis; broiler chickens; surface-exposed antigenic proteins; immune response; gut microbiota.
Online: 28 September 2023 (11:18:47 CEST)
Avian necrotic enteritis is an enteric disease of broiler chickens caused by certain pathogenic strains of Clostridium perfringens in combination with predisposing factors. A vaccine offering complete protection against the disease has not yet been commercialized. In a previous study, we produced five recombinant proteins predicted to be surface-exposed and unique to necrotic enteritis-causing C. perfringens and the immunogenicity of these potential vaccine candidates was assessed in broiler chickens. In the current work, the relative contribution of the antibodies raised by these putative antigens to protect broiler chickens was evaluated using an experimental necrotic enteritis induction model. Additionally, the link between the immune response elicited and the gut microbiota profiles in immunized birds subjected to infection with virulent C. perfringens was studied. ELISA results showed that the IgY antibody titers in vaccinated birds on days 21 and 33 were significantly higher than those on days 7 and 14 and those in birds receiving the adjuvant alone, while the relative contribution of the specific immunity attributed to these antibodies could not be precisely determined using this experimental NE induction model. Besides, 16S rRNA gene amplicon sequencing showed that immunization of birds with recombinant proteins had a low impact on the chicken caecal microbiota.
ARTICLE | doi:10.20944/preprints202309.1945.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: ocular hypertension; intraocular pressure spikes; retinal ganglion cells; inflammasome; caspase-1; interleukin-1β; neuroinflammation
Online: 28 September 2023 (10:02:34 CEST)
Dysfunction and selective loss of retinal ganglion cells (RGCs) is a known cause of vision loss in glaucoma and other neuropathies where ocular hypertension (OHT) is the major risk factor. We investigated the impact of transient non-ischemic OHT spikes (spOHT) on RGC function and viability in vivo to identify cellular pathways linking low-grade repetitive mechanical stress to RGC pathology. We found that repetitive spOHT had an unexpectedly high impact on intraocular homeostasis and RGC viability, while exposure to steady OHT (stOHT) of similar intensity and duration failed to induce pathology. The repetitive spOHT induced a rapid activation of the inflammasome, marked by the upregulation of NLRP1, NLRP3, AIM2, caspases -1, -3/7, -8, gasdermin D (GsdmD) and the release of interleukin-1β (IL-1β) and other cytokines into the vitreous. Similar effects were also detected after 5 weeks of exposure to chronic OHT in an induced glaucoma model. The onset of these immune responses in both spOHT and glaucoma models preceded a 50% deficit in pattern electroretinograms (PERG) amplitude, and significant loss of RGCs at 7 days post-injury. Inactivation of inflammasome complexes in NLRP1-/- , Casp1-/- and GsdmD-/- knockout animals, significantly suppressed the spOHT induced inflammatory response and protected RGCs. Our results demonstrate that mechanical stress produced by acute repetitive spOHT or chronic OHT is mechanistically linked to inflammasome activation, which leads to RGC dysfunction and death.
REVIEW | doi:10.20944/preprints202309.1940.v1
Subject: Biology And Life Sciences, Parasitology Keywords: horse; helminth; cyathostomins; Anoplocephala perfoliata; diagnostics
Online: 28 September 2023 (09:56:56 CEST)
Helminths are commonly found in grazing equids, with the cyathostomin nematodes and the cestode, Anoplocephala perfoliata, being the most prevalent. Most horses harbour low burdens of these parasites and do not develop signs of infection; however, in a small number of animals, high burdens can accumulate and cause disease. Cyathostomins are associated with a syndrome known as larval cyathostominosis. This occurs when large numbers of larvae emerge from the large intestinal wall. This disease has a case fatality rate of up to 50%. A. perfoliata infection has been associated with various types of colic, with burdens of >20 worms associated with pathogenicity. Anthelmintic resistance is a serious problem in cyathostomins and is emerging in A. perfoliata. Control methods that reduce reliance on anthelmintics now need to be applied, especially as no new dewormer compounds are on the horizon. Sustainable control methods must employ diagnostics to identify horses that require treatment. Coprological tests (faecal egg counts, FEC), have been used for several decades to inform treatment decisions to reduce helminth egg shedding. These tests cannot be used to assess host burdens as FEC do not correlate with cyathostomin or A. perfoliata burdens. In the last decade, new tests have become available that measure parasite-specific antibodies, levels of which have been shown to correlate with parasite burden. These tests measure antigen-specific IgG(T) and are available in serum (cyathostomin, A. perfoliata) or saliva (A. perfoliata) formats. Tests for other helminths have been developed as research tools and need to be translated to support equine clinicians in practice. A key element of sustainable control strategies is that diagnostics must be used in combination with management approaches to reduce environmental transmission of helminths; this will help limit the proportion of horses harbouring parasite burdens that need to be targeted by treatment.
BRIEF REPORT | doi:10.20944/preprints202309.1878.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: Horizontal PAGE; Electrode; FIGE
Online: 28 September 2023 (09:45:49 CEST)
The horizontal flatbed electrophoresis system has been used for the electrophoresis of protein samples and is often preferred for a two-dimensional polyacrylamide gel electrophoresis. In currently available equipment, cathode and anode electrodes were placed on top of a gel at each end. An electric field entered from the top of a gel, and its strength was relatively weaker in the bottom part of the gel. Such configuration caused migrating protein bands to lie down forward in the migrating direction. New electrodes, i.e., the clamp-shaped and the double-deck electrodes, were developed to simultaneously apply an electric field from the top and bottom of a gel. Electrophoretic separation with these electrodes made protein bands more focused and improved protein band resolutions. When the FIGE technique was tested together with new electrodes, protein bands were aligned nearly vertically and showed the best electrophoretic resolutions.
ARTICLE | doi:10.20944/preprints202309.1951.v1
Subject: Biology And Life Sciences, Horticulture Keywords: chemical fertilizer; climate change; hormone signaling; plant growth Regulators; abiotic stress; temperature rise; tomato
Online: 28 September 2023 (09:38:04 CEST)
This study evaluated the exogenous application of PGRs substitute chemical fertilization without compromising the growth and yield of tomato in fluctuated day-night temperature and humidity stressed late winter. Two-factor experiment comprising chemical fertilizers at 100, 110, 90 and 80 % of recommended doses besides control and PGRs of GA3; NAA, 4-CPA and SA @ 50 ppm including control was conducted where treatments were assigned in triplicates. Results revealed no significant variation among the fertilizer doses (80% to 110% of recommendation) regarding growth and yield contributing traits while among the PGRs, GA3 @ 50 ppm produced maximum number of flower clusters plant-1 (16.85), flowers (8.80) and fruits (5.79) cluster-1, single fruit weight (67.83 g) and fruit yield (6.61 kg plant-1) of tomato that was statistically identical with the findings of SA. But significant reduction in yield was noted in NAA and 4-CPA (1.20 kg and 1.21 kg plant-1, respectively). Interestingly, GA3 and SA in combination with any doses of the studied fertilizers maximize the tomato morphological and reproductive traits while fertilizer plus NAA and 4-CPA interaction gave the inferior results. Further, correlation matrix and PCA findings revealed that five fertilizer doses have no distinctiveness whereas GA3 and SA has distinct position than other PGRs with the maximum dependent variables those were contributed positively in the total variations. The study findings suggested that 20% fertilizer requirement could be reduced with the substitution of GA3 and SA @ 50 ppm for successful cultivation of tomato in late winter having the extreme environmental issues.
ARTICLE | doi:10.20944/preprints202309.1980.v1
Subject: Biology And Life Sciences, Agricultural Science And Agronomy Keywords: arbuscular mycorrhizal fungi; bio-fertilizer; vetiver; intercropping
Online: 28 September 2023 (09:34:23 CEST)
In this study, a field experiment was conducted to evaluate the growth and yield responses of Sri Lankan lowland rice (Oryza sativa L.) with the application of beneficial Arbuscular mycorrhizal fungi (AMF) inoculum, and inter-cropping with highly mycorrhizal dependent vetiver grass (Chrysopogon zizanioides L.) under two different soil nutrient management systems (NMSs): conventional/chemical (CNMS) and organic (ONMS). The experiment was designed as a split plot with three blocks. Each CNMS and ONMS experiment included untreated control (T0), and three treatments—AMF inoculation (T1), vetiver intercropping (T2), and the combination of AMF and vetiver (T3). According to the results, colonization of rice roots with AMF was not affected significantly by the treatments and ranged from 0‒15.8%. The effect was very low or absent in the early stage and then higher in the later stages of the rice plant. Furthermore, plant growth was not significantly different between the two NMSs, although grain yield was significantly higher (P < 0.05), with the order T1 (0.45 kg/m2) >T2 (0.42 kg/m2) >T3 (0.41 kg/m2) in CNMS and T2 (0.44 kg/m2) >T1 (0.41 kg/m2) >T3 (0.40 kg/m2), in ONMS than for the respective controls (T0), thus suggesting beneficial utilization of AMF and vetiver in the lowland rice farming system.
REVIEW | doi:10.20944/preprints202309.1979.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: autoimmunity; glycolysis; metabolites; psoriasis; rheumatoid arthritis; T-cell function; T-cell motility; T-lymphocytes
Online: 28 September 2023 (09:01:58 CEST)
Immune surveillance and adaptive immune responses, involving continuously circulating and tissue-resident T-lymphocytes, provide host defense against infectious agents and possible malignant transformation while avoiding autoimmune tissue damage. Activation, migration, and deployment of T-cells to affected tissue sites are crucial for mounting an adaptive immune response. An effective adaptive immune defense depends on the ability of T-cells to dynamically reprogram their metabolic requirements in response to environmental cues. Inability of the T-cells to adapt to specific metabolic demands may skew cells to become either hyporesponsive creating immunocompromised conditions or hyperactive causing autoimmune tissue destruction. Here, we review maladaptive T-cell metabolic fitness that can cause autoimmune diseases and discuss how T-cell metabolic programs can potentially be modulated to achieve therapeutic benefits.
REVIEW | doi:10.20944/preprints202309.1925.v1
Subject: Biology And Life Sciences, Agricultural Science And Agronomy Keywords: cotton; irrigation; fertilization; water deficit; efficiency; productivity
Online: 28 September 2023 (08:37:47 CEST)
The limited availability of water resources worldwide presents obstacles, to the progress of ag-riculture. This situation calls for the implementation of water saving technologies to address these challenges effectively. The quest for technologies that conserve water in agriculture, par-ticularly within the context of cotton production, is related to the study of the impact of drought on the growth, leaf surface area, accumulation of dry matter, boll number, and productivity. The benefits of local drip irrigation have been established. Various models and strategies for opti-mizing and improving the efficiency of how we use water for irrigation purposes. In a market driven economy the effective utilization of water assetsis closely linked to the improvement of managing mineral nutrients. To enhance both the quantity and quality of production, nitrogen is the key factor element that plays a role in the process. The formation of highly efficient cotton crops by adapting the growing technology to the specific soil-climatic conditions of the area, in-cluding controlling nitrogen fertilizing and irrigation, is the primary method to achieve con-sistent harvests. The results of irrigation with water deficit and feeding plants with different fer-tilizer rates are presented. This matter is presently significant, in achieving dependable crop harvests supporting the economic growth of farms and safeguarding the well being of agricul-tural environments.
ARTICLE | doi:10.20944/preprints202309.1760.v1
Subject: Biology And Life Sciences, Agricultural Science And Agronomy Keywords: nitrogen source; food security; cyanobacteria; Arthrospira maxima; phycobiliproteins.
Online: 28 September 2023 (05:30:13 CEST)
Limnospira maxima is widely cultivated due to its abundance of proteins, carbohydrates, vitamins, minerals, essential fatty acids, and phycobiliproteins. However, limited information, high production costs, and nitrogen source limitations have hindered the optimal utilization of this cyanobacteria. The main goal of this study was to address these limitations by identifying ideal cultivation parameters to maximize yields, reduce costs, and contribute to global food security. It was demonstrated that the white and yellow light spectra are the most suitable for cultivating L. maxima, and the use of KNO3 as a nitrogen source proved to be the most cost-effective on a large scale. Nitrogen deficiency stimulates the production of dry biomass but also affects the concentration of key pigments such as chlorophyll and phycocyanin. A notable characteristic of L. maxima is its adaptability and survival capability in various environmental conditions, including nitrogen-deficient environments. This resilience is achieved through the degradation of phycobilisomes and the storage of glycogen. By identifying the optimal growth conditions and understanding the physiological responses of L. maxima, cultivation practices can be improved to be more efficient and sustainable.
ARTICLE | doi:10.20944/preprints202309.1906.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Iberian pig; litter size; inbreeding depression
Online: 28 September 2023 (03:36:29 CEST)
Inbreeding depression is expected to be more pronounced in fitness-related traits, such as pig litter size. Recent studies have suggested that the genetic determinism of inbreeding depression may be heterogeneous across the genome. Therefore, the objective of this study is to conduct a genomic scan across the pig autosomal genome to detect the genomic regions that control inbreeding depression for litter size in two varieties of Iberian pigs (Entrepelado and Retinto). The datasets consist of 2,069 (338 sows) and 2,028 (327 sows) records for litter size (Total Number Born and Number Born Alive) for the Entrepelado and Retinto varieties. All sows were genotyped using the Geneseek GGP PorcineHD 70 K. We employed the Unfavorable Haplotype Finder software to extract runs of homozygosity (ROHs) and conducted a mixed model analysis to identify highly significant differences between homozygous and heterozygous sows for each specific ROH. A total of 8 genomic regions located on SSC2, SSC5, SSC7, SSC8, and SSC13, were significantly associated with inbreeding depression, housing some relevant genes such as FSHR, LHCGR, CORIN, AQP6, and CEP120.
ARTICLE | doi:10.20944/preprints202309.1888.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: antidiabetic, anti-inflammatory activity, Artemisia absinthium, Artemisia vulgaris, Trigonella foenum-graecum, membrane technologies, α-amylase, α-glucosidase, hyaluronidase, lipoxygenase
Online: 28 September 2023 (03:23:11 CEST)
In recent years there has been increasing interest in the discovery of new natural herbal remedies for the treatment of common conditions such as diabetes and inflammatory diseases. In this context, this paper analyzed the antidiabetic and anti-inflammatory potential of two plants of the genus Artemisia: Artemisia absinthium (wormwood) and Artemisia vulgaris (mugwort) and Trigonella foenum-graecum (fenugreek) plants less studied from this point of view. Thus, hydroal-coholic extracts (50% EtOH v/v, 10% mass) were made, which were processed by membrane technologies, micro- and ultrafiltration, in order to concentrate the biologically active principles. The extracts were then analyzed in terms of content in active principles of interest (polyphenols, flavones). The antidiabetic activity of the extracts was analyzed by testing their ability to inhibit α-amylase and α-glucosidase and the anti-inflammatory activity by testing the ability to inhibit hyaluronidase (HYA) and lipoxygenase (LOX). Thus, the T. foenum-graecum extracts showed a high inhibitory activity on α-amylase 3.22 ± 0.3 g / mL (compared to the standard used - IC50 acarbose = 3.5 g / mL) and a high inhibitory activity on LOX: 19.69 ± 0.52 g/mL (compared to all standards used). The concentrated extract of Artemisia vulgaris showed increased activity of inhibition of α-amylase - IC50 = 8.57 ± 2.31g/mL and moderate activity of inhibition of HYA - IC50 = 17.18 g/mL compared to ibuprofen, the standard used - IC50 = 5.73 g/mL. The concentrated extract of Artemisia absinthium showed a pronounced activity of inhibition of LOX - IC50 = 19.71 g / mL, compared to the standard - rutin - IC50 = 22.34 g/mL).
ARTICLE | doi:10.20944/preprints202309.1902.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: embryonic development; temperature; autophagy; apoptosis; Sepiella japonica
Online: 28 September 2023 (03:09:41 CEST)
Temperature is a crucial environmental factor that affects embryonic development, particularly for marine organisms with long embryonic development periods. However, the sensitive period of embryonic development and the role of autophagy/apoptosis in temperature regulation in cephalopods, enigmatic creatures, remain unclear. In this study, we cultured embryos of Sepiella japonica, a typical species in the local area of the East China Sea, at different incubation temperatures (18 ℃, 23 ℃, and 28 ℃) to investigate various developmental aspects, including morphological and histological characteristics, mortality rates, the duration of embryonic development, and expression patterns of autophagy-related genes (LC3, BECN1, Inx4) and apoptosis marker genes (Cas3, p53) at 25 developmental stages. Our findings indicate that embryos in the high-temperature (28 ℃) group had significantly higher mortality and embryonic malformation rates than those in the low-temperature (18 ℃) group. Furthermore, high temperature (28 ℃) shortened the duration of embryonic development by 7 days compared to the optimal temperature (23 ℃), while low temperature (18 ℃) caused a delay of 9 days. Therefore, embryos of S. japonica were more intolerant to high temperatures (28 ℃), emphasizing the critical importance of maintaining an appropriate incubation temperature (approximately 23 ℃). Additionally, our study observed, for the first time, that the Early blastula, Blastopore closure, and Optic vesicle to Caudal end stages were the most sensitive stages. During these periods, abnormalities in the expression of autophagy-related and apoptosis-related genes were associated with higher rates of mortality and malformations, highlighting the strong correlation and potential interaction between autophagy and apoptosis in embryonic development under varying temperature conditions.
ARTICLE | doi:10.20944/preprints202309.1891.v1
Subject: Biology And Life Sciences, Endocrinology And Metabolism Keywords: thermography; tendons; quadriceps muscle; body temperature regulation
Online: 28 September 2023 (03:09:33 CEST)
In the last decade there has been a growing interest in infrared thermography in the field of sports medicine in order to elucidate the mechanisms of thermoregulation. The aim of this study was to describe bilateral variations in skin temperature of the anterior thigh and patellar tendon in healthy athletes, to provide a model of baseline tendon and muscle thermoregulation in healthy sprinters following a unilateral isokinetic fatigue protocol. Fifteen healthy national-level sprinters underwent unilateral isokinetic force testing and electrostimulation in which body temperature was measured before, during, and after the protocol using an infrared thermographic camera. ANOVA detected a significant difference in the time x side interaction for patellar temperature changes (p≤0.001) and a significant difference in the time x side interaction for quadriceps temperature changes (p≤0.001). The thermal challenge produces homogeneous changes evident in quadriceps areas, but not homogeneous in tendon areas. these data show that metabolic and blood flow changes may depend on the physical and mechanical properties of each tissue.
BRIEF REPORT | doi:10.20944/preprints202309.1856.v1
Subject: Biology And Life Sciences, Horticulture Keywords: strawberry cultivars; a sum of effective temperatures; K-means clustering; phenological growth stages
Online: 28 September 2023 (03:04:38 CEST)
Finding the ideal statistical method for grouping phenological data is always an important step for breeders to draw correct conclusions from it possibly. In this paper, K-cluster analysis is presented as a perfect tool for grouping phenological data. The present research was performed based on the phenological data of 61 strawberries (Fragaria × ananassa) cultivars of different geo-graphical origins grown in Estonian conditions. Groups of strawberry cultivars were deter-mined according to flowering and ripening time: early, middle and late, based on the sum of ef-fective temperatures above +5°C. The result of the K-cluster analysis carried out in this way makes it possible to precisely plan the ripening time of berries of different strawberry cultivars. Using such analysis data, it is possible to combine with different early, mid or late strawberry cultivars to extend the picking period. Also, this technique can be used to study the effect of cli-matic changes occurring over the years on the phenology of strawberry cultivars grown in the region.
ARTICLE | doi:10.20944/preprints202309.1813.v1
Subject: Biology And Life Sciences, Plant Sciences Keywords: heat stress; nicotine biosynthesis; radiocarbon flux analysis; carbon-11; protein degradation; L-Asparaginase activity
Online: 28 September 2023 (02:50:22 CEST)
The effect of high temperature (HT) stress on nicotine biosynthesis in Nicotiana attenuata was examined. Nicotine content was measured in mature leaves, young sink leaves and in roots from well-watered plants grown at 25 °C as controls and from plants exposed to 38 °C and 43 °C temperatures applied for 24, 48, 72 and 96-hr duration. At 38 °C, all leaf nicotine levels were significantly less than control plants for up to 72-hr exposure but rose sharply thereafter to levels significantly greater than controls with 96-hr exposure. In contrast, plants exposed to 43 °C never exhibited a reduction in leaf nicotine content and showed a significant increase in content with just 48-hr exposure. When radioactive 11CO2 was administered to plants, HT stress significantly reduced fixation. Furthermore, radiocarbon flux analysis revealed that ‘new’ C partitioning (as 11C) into the [11C]-amino acid pool was significantly reduced with HT stress as were yields of [11C]-aspartic acid, an important amino acid in nicotine biosynthesis, and its beta-amido counterpart [11C]-asparagine significantly reduced. In contrast, [12C]-aspartic acid levels appeared unaffected at 38 °C but were significantly elevated at 43 °C relative to controls. Additionally, [12C]-asparagine levels were significantly elevated at both stress temperatures. Finally, leaf total soluble protein (TSP) was reduced 39% with long exposures to both stress temperatures. However, Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) which was 41% TSP appeared unaffected by HT stress. Altogether, these results support the theory that plant proteins other than Rubisco degrade at elevated temperatures freeing up essential amino acids supporting the nicotine biosynthetic machinery, and while feedback might downregulate de novo amino acid synthesis from ‘new’ C resources, more than likely plant C/N rebalancing caused by HT reductions in C input restricted N uptake, which in turn, limited this process.
REVIEW | doi:10.20944/preprints202309.1841.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: tuberculosis; Mycobacterium; physiology; mycolic acids; cell wall
Online: 28 September 2023 (02:45:57 CEST)
Mycobacterium tuberculosis is the agent of tuberculosis, one of the most important infectious diseases in the world. This microorganism stands out from other bacteria, not only for its extremely high infection capacity, but also for its cellular characteristics that include an extremely resistant and hydrophobic cellular parade the passage of antibiotics. An incredible ability to adapt to adverse conditions inside the host as well as its vast arsenal of virulence factors that allow its survival within the inhospitable environment within the macrophage can be highlighted. This review aims to discuss several aspects of MTB microbiology, genetics, and physiology. We will address in this review details of the metabolism of MTB that allows it to replicate in the active phase and remain viable during latency, as well as the characteristics of its cell wall that contribute to the blockade of the immune response and its resistance to antibiotics.
REVIEW | doi:10.20944/preprints202309.1897.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: subiculum; wake; sleep; REM sleep; NREM sleep; vigilance state
Online: 27 September 2023 (14:29:31 CEST)
This review provides an overview of the current understanding of the neural systems involved in regulating wakefulness, non-rapid eye movement sleep (NREMS), and rapid eye movement sleep (REMS) in mammals. Specifically, we focus on the anatomical connections between the subiculum, a component of the hippocampal formation, and the regions responsible for regulating the sleep-wake cycle. The subiculum exhibits direct connections with key areas involved in sleep regulation, such as the lateral hypothalamus, tuberomammillary nucleus, basal forebrain, ventrolateral preoptic nucleus, ventrolateral tegmental area, and suprachiasmatic nucleus. Additionally, second-order projections from the subiculum are received by the laterodorsal tegmental nucleus and locus coeruleus, suggesting potential involvement of the subiculum in the regulation of circadian rhythms, particularly the circadian sleep-wake cycle. We also discuss alterations in the subiculum observed in individuals with sleep disorders and sleep-deprived mice, underscoring the significance of investigating neuronal communication between the subiculum and pathways promoting both sleep and wakefulness.
ARTICLE | doi:10.20944/preprints202309.1871.v1
Subject: Biology And Life Sciences, Other Keywords: cognitive flexibility; mood; energy; concentration; motivation; nootropics
Online: 27 September 2023 (11:52:54 CEST)
This study assessed the acute effects of oral methylliberine (DynamineTM) supplementation on cognitive function and indices of well-being. This was a double-blind, randomized, within-subject crossover trial. 25 healthy men and women (33.5±10.7yr, 172.7±8.6cm, 73.3±11.0kg) underwent pretesting before ingesting methylliberine (100mg) or placebo (PLA) for 3 days. On the fourth day participants were tested before their 4th dose (baseline) and every hour post-ingestion for 3 hours. After a one-week washout period, participants repeated testing with the alternate investigational product. The testing battery consisted of vitals, Stroop test and Trail Making Test-B, and visual analog scales that assessed various indices of well-being. Mixed factorial ANOVA with repeated measures were used to assess all variables. There were significant (p≤0.050) interactions in concentration, motivation, and mood. Methylliberine improved concentration 1- and 3-hours, motivation 3 hours, and mood 1-, 2-, and 3 hours (p≤0.050). Methylliberine improved energy, sustained energy, and mood in all participants to a greater extent than PLA at 1-hour and 3-hours relative to baseline (p≤0.050). PLA improved motivation at 1- and 2-hours and mood at 2-hours (p≤0.050). Methylliberine improved concentration, well-being, and the ability to tolerate stress to a greater extent than PLA at 3-hours relative to baseline (p≤0.050). Women observed elevations in sustained energy 1- and 3-hours (p≤0.050) in methylliberine vs. PLA. Methylliberine had negligible influence on cognitive function, vitals (p>0.050), and no adverse events were reported. Methylliberine significantly improved subjective feelings of energy, concentration, motivation, and mood, but not cognitive function. PLA improved motivation and mood at hours 1 and 2, while methylliberine sustained benefits for longer. Methylliberine also improved concentration, well-being, and the ability to tolerate stress to a greater degree than PLA while having no detrimental effects on vital signs. Methylliberine also seems to have a positive impact on sustained energy in women.
REVIEW | doi:10.20944/preprints202309.1893.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: atherosclerosis; coronary artery disease; stent; bio-resorbable; 3D printing
Online: 27 September 2023 (11:23:16 CEST)
Coronary artery disease is the most prevalent cardiovascular disease, claiming millions of lives annually around the world. The current treatment includes surgically inserting a tubular construct called as a stent inside arteries to restore blood flow. However, due to lack of patient-specific design, the commercial products cannot be used with different vessel anatomies. In this review, we have summarized the drawbacks in existing commercial metal stents which face problems of restenosis and inflammatory responses, owing to the development of neointimal hyperplasia. Further, we have highlighted the fabrication of stents using biodegradable polymers, which can circumvent most of the existing limitations. In this regard, we elaborated on utilization of new fabrication methodologies based on additive manufacturing such as three-dimensional printing to design patient-specific stents. Finally, we have discussed on the functionalization of these stent surfaces with suitable bioactive molecules which can prove to enhance their properties in preventing thrombosis and better healing of injured blood vessel lining.
ARTICLE | doi:10.20944/preprints202309.1870.v1
Subject: Biology And Life Sciences, Aquatic Science Keywords: stress; gonad; sex; methylation; masculinization; rearing
Online: 27 September 2023 (10:46:16 CEST)
Rearing density directly impacts fish welfare, which, in turn, affects productivity in aquaculture. Previous studies have indicated that high-density rearing during sexual development in fish can induce stress, resulting in a tendency towards male-biased sex ratios in the populations. In recent years, research has defined the relevance of the interactions between the environment and epigenetics playing a key role in the final phenotype. However, the underlying epigenetic mechanisms of individuals exposed to confinement remain elucidated. By using zebrafish (Danio rerio) the DNA methylation promotor region and the gene expression patterns of six genes: dnmt1, cyp19a1a, dmrt1, cyp11c1, hsd17b1, and hsd11b2 involved in the DNA maintenance methylation, reproduction, and stress were assessed. Zebrafish larvae were subjected to two high-density conditions (9 and 66 fish/L) during two periods of overlapping sex differentiation of this species (7 to 18 and 18 to 45 days post-fertilization, dpf). Results showed a significant masculinization in the populations of fish subjected to high densities from 18 to 45 dpf. In adulthood the dnmt1 gene was differentially hypomethylated in ovaries and its expression was significantly downregulated in the testes of fish exposed to high-density. Further, the cyp19a1a gene showed downregulation of gene expression in the ovaries of fish subjected to elevated density, as previously observed in other studies. We proposed dnmt1 as a potential testicular epimarker, and the expression of ovarian cyp19a1a as a potential biomarker for predicting stress originated from high densities during the early stages of development. These findings highlight the importance of rearing densities by long-lasting effects in adulthood conveying cautions for stocking protocols in fish hatcheries.
ARTICLE | doi:10.20944/preprints202309.1659.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Tendinopathy; tendon lesion; mesenchymal stem cells; mesenchymal progenitor cells; MSC; adipose; horse serum; clinical study; re-injury; recurrence rate
Online: 27 September 2023 (10:43:19 CEST)
Treatment of tendinopathies with multipotent mesenchymal stromal cells (MSCs) is a promising option in equine and human medicine. However, conclusive clinical evidence is lacking. The purpose of this study was to gain insight into clinical treatment efficacy and to identify suitable outcome measures for larger clinical studies. Fifteen horses with early naturally occurring tendon disease were assigned to intralesional treatment with allogeneic adipose-derived MSCs suspended in serum or with serum alone by block randomization (dosage adapted to lesion size). Clinicians and horse owners remained blinded to the treatment during 12 months (7 horses per group) and 18 months (7 MSC- and 5 control-group horses) of follow-up including clinical examinations and diagnostic imaging. Clinical inflammation-, lameness- and ultrasonography scores improved more over time in the MSC-group. The lameness score difference significantly improved in the MSC- compared to the control-group after 6 months. In the MSC-group, 5 out of 7 horses were free of re-injuries and back to training until 12 and 18 months. In the control-group, 3 out of 7 horses were free of re-injuries until 12 months. These results suggest that MSCs are effective for treatment of early phase tendon disease and provide a basis for a larger controlled study.
ARTICLE | doi:10.20944/preprints202309.1874.v1
Subject: Biology And Life Sciences, Other Keywords: dehydration; drinking pattern; exercise; heat stress; physiological and perceptual functions
Online: 27 September 2023 (10:36:15 CEST)
A drinking strategy aiming to replace a given percentage of the sweat losses incurred during exercise should result in reproducible fluid intake volume and, hence, fluid balance from one exercise session to the other performed under similar scenarios. Whether this may also be the case with ad libitum drinking during exercise is unclear. We characterized the repeatability of ad libitum fluid intake during repeated 1 h exercise sessions and examine its effect over time on fluid balance and selected physiological functions and perceptual sensations. Twelve (3 women) healthy individuals participated in this study. At weekly intervals, they completed 4, 2 x 30 min walking/jogging exercise bouts (55% VO2max, 40°C, 20-30% relative humidity) interspersed by a 3 min recovery period. During exercise, participants replaced water (20°C) ad libitum. There were no significant differences among the 4 exercise sessions for absolute fluid intake volume (~ 1000 mL · h-1), percent body mass loss (~ 0.4%), sweat rate (~ 1300 mL · h-1) and percent of sweat loss replaced by fluid intake (~ 80%). Heart rate, rectal temperature and perceived thirst and heat stress did not differ significantly between the first and fourth exercise session. Perceived exertion was significantly lower during the fourth vs. the first exercise session, but the difference was trivial, < 1 arbitrary unit. In conclusion, ad libitum fluid intake during 4 successive identical 1 h walking/jogging sessions conducted in the heat will result in similar fluid intake volumes and perturbations in fluid balance, heart rate, rectal temperature and perceived thirst, heat stress and exertion.
ARTICLE | doi:10.20944/preprints202309.1866.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: the celestial goldfish; blood; vitreous humor; physiological indicators; biochemical indicators
Online: 27 September 2023 (10:18:49 CEST)
The aim of this study was to examine and understand the physiological and biochemical components of the blood and vitreous humor of goldfish (Carassius auratus). Blood was drawn from the arterial (static) line and vitreous humour was extracted using a syringe from healthy goldfish. An automatic haematology analyzer was used to detect 17 physiological indicators, including white blood cells and red blood cells. At the same time, 20 biochemical indexes, including albumin, calcium, and glutamic pyruvic transaminase, were found in the automatic biochemical analyzer. Experiments were also conducted on the effect of blood and vitreous fluid collection on the subsequent survival rate of the celestial goldfish. The findings demonstrated that there were values for 17 physiological indicators in the blood of the goldfish, including red blood cells (2.19 × 1012), white blood cells (62.21 × 109), hemoglobin (138.25g/L), and no eosinophils or basophils. However, no data for 17 physiological indications of vitreous humor were discovered. The quantities of total protein, glutamic oxaloacetic transaminase, and potassium in the vitreous humor were substantially lower than those in the serum (P < 0.01) according to the results of 20 biochemical indexes in the serum and vitreous humor of goldfish. Alkaline phosphatase, chlorine, and creatinine between vitreous humor and serum did not differ significantly(P > 0.05). The levels of glutamic pyruvic transaminase and γ-glutamyl transpeptide in vitreous humor were significantly higher than those in serum (P < 0.01). This experiment provides basic datas for the healthy culture of goldfish and the perfection of hematology in goldfish, and the results will be convenient for the further study of development and formation of celestial eye traits.
ARTICLE | doi:10.20944/preprints202309.1869.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Enterocins; Enterococcus; breast-fed infants; bacterial infection; Listeria monocytogenes; selective inhibition; probiotic
Online: 27 September 2023 (09:17:31 CEST)
Enterocins are bacteriocins synthesized by Enterococcus strains that show an interesting antimi-crobial effectiveness against foodborne pathogens such as Listeria monocytogenes. The objectives of this study were identify and analyze the expression of enterocin genes of Enterococcus isolated from breast-fed infants and evaluate the ability to inhibit three human isolates of virulent Lis-teria monocytogenes, as well as some probiotic bacteria. The susceptibility of the strains of L. mon-ocytogenes to fifteen antibiotics was tested, detecting resistance to cefoxitin (constitutive-ly resistant), oxacillin and clindamycin. Production of enterocins A, B and P were observed in Enterococcus faecium isolates, and enterocin AS-48 in an Enterococcus faecalis isolate. AS-48 showed antilisterial activity by itself, while the joint action of enterocins A and B, or B and P was neces-sary for inhibiting L. monocytogenes, demonstrating a synergistic effect of those combinations. The presence of multiple enterocin genes does not assured the inhibition of L. monocytogenes strains. However, the expression of multiple enterocin genes showed a good correlation with the inhibition capacity of these strains. Furthermore the potential beneficial strains of lactobacil-li and bifidobacteria examined were not inhibited by any of the enterocins produced individu-ally or in combination, with the exception of Bifidobacterium longum BB536, which was inhibited by enterocin AS-48 and the joint production of enterocins A and B or B and P. The enterocins studied here could be candidates for developing alternative treatments against antibi-otic-resistant bacterial infections. Moreover, these selected enterocin-producing E. faecium strains isolated from breast-fed infants could be used as probiotic strains due to their antilisterial effect as well as the absence of virulence factors.
ARTICLE | doi:10.20944/preprints202309.1837.v1
Subject: Biology And Life Sciences, Food Science And Technology Keywords: Lemon myrtle; essential oil; citral; citrus fruit; rind injury; Penicillium digitatum; sensory test
Online: 27 September 2023 (07:20:57 CEST)
This study examined the efficacy of essential oil (EO) obtained from the Australian native plants, lemon myrtle (Backhousia citriodora) (LM) to inhibit Penicillium digitatum by in vitro agar diffusion and vapor assay, and in inoculated oranges. Gas chromatograph/mass spectrometry analysis revealed the main constituent of LM EO as citral. Pure citral was also included in the experiment for com-parison. LM EO at 1, 2, 3, 4 and 5 µL disc-1 in the in vitro tests significantly inhibited fungal growth compared to the non-treated control. Moreover, LM EO at 4 and 5 µL disc-1 in both the assays completely inhibited fungal growth similar to its main component of citral. Inoculated oranges dipped in 1000 μL L-1 LM EO solutions for 5, 10, 15 and 30 sec showed significantly lower fungal wounds compared to control. While higher dipping times generated slight rind injury, a 5 and 10 sec dip did not cause any injury to the fruit rind. The quality assessments and sensory evaluations also revealed that the addition of LM EO did not adversely affect the quality and taste of the oranges. These findings suggest LM EO as an alternative to synthetic fungicides to inhibit wastage in citrus during storage.
ARTICLE | doi:10.20944/preprints202309.1816.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: phosphate-accumulating microorganisms; EBPR; Ca. Accumulibacter; oxygen uptake
Online: 27 September 2023 (06:59:11 CEST)
Candidatus Accumulibacter belongs to phosphate-accumulating organisms (PAO) which exhibit cyclic metabolism and are capable of intracellular polyphosphate accumulation and their hydrolysis under feast-famine anaerobic-aerobic cycling. In consortia of activated sludge microorganisms, these bacteria are responsible for enhanced biological phosphorus removal (EBPR). The spectrum of the substrates used by Ca. Accumulibacter remains insufficiently studied. It was investigated by measuring the oxygen uptake rates (OUR) of Ca. Accumulibacter-enriched culture supplemented with 17 different organic substrates. The highest oxygen uptake rates values were observed in the presence of tryptone, volatile fatty acids (acetate, propionate, and butyrate), succinate, pyruvate, and amino acids (aspartate and glutamate). Phosphate dynamics in the medium under shifts from anaerobic to aerobic cultivation in batch experiments was studied for these compounds (except for tryptone). All tested substrates were shown to cause phosphate cycling (release in the anaerobic phase and uptake in the aerobic one), with OURs for the substrates correlating with the amount of phosphates consumed during the aerobic phase. It was concluded that OUR may be used as an indicator of the monosubstrates used by Ca. Accumulibacter in the anaerobic/aerobic cycle. The possible pathways for substrate transport and metabolism by Ca. Accumulibacter are discussed using the stoichiometric data and the results of metagenomic analysis.
ARTICLE | doi:10.20944/preprints202309.1839.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: Blue light; Oxidative stress; Pigmentation; Tocotrienol-rich fraction; Antioxidant; Anti-melanogenesis; Melanocyte
Online: 27 September 2023 (06:32:14 CEST)
Our skin is constantly exposed to blue light (BL), which is abundant in sunlight and emitted by digital devices. Prolonged exposure to BL can lead to oxidative stress-induced damages and skin hyperpigmentation. In this study, we investigated the protective effects of tocotrienol-rich fraction (TRF) on BL-induced oxidative stress and hyperpigmentation in B16-F1 melanocytes. Alpha-tocopherol (αTP) was used as a comparator. Our results showed that TRF effectively suppressed the formation of reactive oxygen species, and preserved the mitochondrial membrane potential. Additionally, TRF exhibited anti-apoptotic properties by reducing the activation of p38 mitogen-activated protein kinase molecule and downregulating the expression of cleaved caspase-3. Moreover, TRF modulated tyrosinase activity, resulting in a lowered rate of melanogenesis and reduced melanin production. In contrast, αTP did not exhibit the significant protective effects against skin damages and pigmentation in BL-induced B16-F1 cells. Therefore, this study indicates that TRF may offer superior protective effects over αTP against the effects of BL on melanocytes. These findings demonstrate the potential of TRF as a protective natural active against BL-induced skin damages and hyperpigmentation via its antioxidative and anti-melanogenic properties.
ARTICLE | doi:10.20944/preprints202309.1838.v1
Subject: Biology And Life Sciences, Food Science And Technology Keywords: CBD; CBDA; cannabis sativa; hemp oil, hemp flowers, hemp stalks, rheology; protein dough.
Online: 27 September 2023 (06:03:43 CEST)
(1) Background: In recent years, the increasing demand for alternative food shifted research toward new types of food sources. It is well known that many diseases are caused by oxidative stress, then supplementation of antioxidants can be proposed to reduce it. Cannabis Sativa (CS) is an interesting alternative source of antioxidant species. The use of CS for the extraction of these molecules could be optimized to maximize the antioxidant yield extracts. (2) Methods: This work aimed to investigate extraction methods of CBD oil from CS inflorescences and stalks to obtain antioxidant-enriched compounds for use in food by mixing with a carrier oil like hemp seed oil. Soxhlet (SE), dynamic maceration (DME) and supercritical fluid (SFE) extraction techniques were performed, using CS inflorescences and stalks; by High-Pressure Liquid Chromatography (HPLC) analysis the antioxidants were individuated and quantified; moreover, the antioxidant power was investigated by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays and finally doughs enriched with hemp oil were studied by rheological measurements (3) Results: The best results in terms of CBD yield were obtained by dynamic maceration from only flowers and it also shows a good antioxidant power. (4) Conclusions: Formulation of protein bars with hemp oil enriched in CBD could be an excellent means for the consumption of products enriched with antioxidants because it preserves the CBD anti-inflammatory activity.
DATA DESCRIPTOR | doi:10.20944/preprints202309.1802.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: PAD4/PADI4; PAD2/PADI2; Interstitial Lung Disease; Rheumatoid Arthritis; RA-ILD, AIM
Online: 27 September 2023 (05:56:40 CEST)
Rheumatoid arthritis (RA) is an autoimmune disease mainly characterized by joint inflammation. It presents extra-articular manifestations, with the lungs one of the affected areas. Among these, damage to the pulmonary interstitium (Interstitial Lung Disease -ILD) has been linked to proteins involved in the inflammatory process and related to extracellular matrix deposition and lung fibrosis establishment. Peptidyl arginine deiminase enzymes (PAD), which carry out protein citrullination, play a role in this context. A genetic association analysis was conducted on genes encoding two PAD isoforms, PAD2 and PAD4. This analysis also included ancestry informative markers and protein level determination in samples from patients with rheumatoid arthritis, RA-associated ILD, and clinically healthy controls. Significant single nucleotide variations (SNV) and a haplotype were identified as susceptibility factors for ILD-RA development. Elevated levels of PAD4 were found in ILD-RA cases, while PADI2 showed an association with RA susceptibility. This document presents the data obtained from the conducted research, which has been published.
ARTICLE | doi:10.20944/preprints202309.1836.v1
Subject: Biology And Life Sciences, Cell And Developmental Biology Keywords: adipose stromal cells; spheroids; chondrogenesis; mass density; deep imaging
Online: 27 September 2023 (05:23:08 CEST)
Articular cartilage lacks intrinsic regenerative capabilities, and the current treatments fail to regenerate damaged tissue and lead only to temporary pain relief. These limitations have prompted the development of tissue engineering approaches, including 3D culture systems. Thanks to their regenerative properties and capacity to recapitulate embryonic processes, spheroids obtained from mesenchymal stromal cells are increasingly studied as building blocks to obtain functional tissues. The aim of this study was to investigate the capacity of adipose stromal cells to assemble in spheroids and differentiate toward chondrogenic lineage from the perspective of cartilage repair. Spheroids were generated by two different methods (3D chips vs Ultra-Low Attachment plates), differentiated toward chondrogenic lineage, and their properties were investigated by: molecular biology analyses, biophysical measurement of mass density, weight, and size of spheroids, and confocal imaging. Overall, spheroids showed the ability to differentiate by expressing specific cartilaginous markers that correlate with their mass density, defining a critical point at which they start to mature. Considering the spheroid generation method, this pilot study suggested that spheroids obtained with chips are a promising tool for the generation of cartilage organoids that could be used for preclinical/clinical approaches, including the realization of personalized therapy.
ARTICLE | doi:10.20944/preprints202309.1829.v1
Subject: Biology And Life Sciences, Horticulture Keywords: yellow sapote; tree selection; viviparity; leaf characterization; early seed germination
Online: 27 September 2023 (05:13:32 CEST)
Canistel (Pouteria campechiana (Kunth) Baehni) (yellow sapote, canistel). Originally from Mexico, the fruit is edible, and the tree is used as an ornamental and medicinal plant. Descriptive studies were carried out with the objective of morphologically characterizing the fruits and leaves of trees located in different regions of Mexico to select outstanding specimens and propose their massive reproduction. The trees were selected in three zones. In zone 2 from flower anthesis to harvest 11 months, fruits with 3 seeds, greater weight (180 to 330 g) and quantity of pulp (198.88 g), subglobose shape, total soluble solids 33 %. The fruits of zone 1 at harvest 8 months and zone 3 took 9 months to be harvested, in the rest of the characteristics evaluated were exceeded by the fruits of zone 2. To characterize the canistel (P. campechiana) leaf and identify accessions or cultivars, the three main variables, according to the principal component analysis were: leaf area, perimeter, and minor diameter. The three zones where canistel fruits and leaves were characterized are important and can be recommended for propagation in zones with favorable climatic conditions for fruiting in tropical and subtropical regions of Mexico.
ARTICLE | doi:10.20944/preprints202309.1808.v1
Subject: Biology And Life Sciences, Virology Keywords: HIV-1; HERV-K; elite controllers; viremic controllers; restriction factors
Online: 27 September 2023 (05:09:21 CEST)
HIV-1 infection can activate the expression of human endogenous retroviruses (HERVs), particularly HERV-K (HML-2). HIV controllers (HICs) are rare people living with HIV (PLWH) who naturally control HIV-1 replication. The ability of HICs to control the expression of endogenous retroviruses has not been previously addressed. In this study, we measured ERVK-6 RNA expression in PBMCs of HICs (n = 23), antiretroviral (ART)-suppressed subjects (n = 8), and HIV-1-negative (NEG) individuals (n = 10) and correlated the transcript expression of ERVK-6 with multiple HIV-1 restriction factors. Our study reveals that ERVK-6 RNA expression in PBMCs from HICs was significantly downregulated compared with both ART and NEG control groups. Moreover, we detected that ERVK-6 RNA levels in PBMCs across all groups were negatively correlated with the expression levels of p21 and MCPIP1. In our previous study, these two cellular restriction factors were upregulated in HICs compared with control groups. Interestingly, p21 and MCPIP1 limit the activation of macrophages and T cells by downregulating the activity of NF-kB, a transcription factor that stimulates the LTR-driven transcription of HIV-1 and HERV-K proviruses. These findings support that HICs activate innate antiviral mechanisms that may simultaneously downregulate the transcription of both HIV-1 and endogenous retroviruses.
REVIEW | doi:10.20944/preprints202309.1791.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: baking; yeast; LAB; sourdough; essential oils; plant extracts; probiotics; drying preservation
Online: 27 September 2023 (04:25:46 CEST)
The required processes and steps for making bread include technological and innovative concepts. The current trend is the use of less toxic compounds and green processes. Besides lactic acid bacteria and yeast, other microorganisms with special properties, such as enzymes, new aroma and flavor, exopolysaccharides, and vitamins, among other compounds with beneficial properties, could be added to bread manufacture, improving bread quality and health effects for the consumers. Freeze drying, new encapsulation methods, cryoprotectants, spray drying, fluidized bed drying, and vacuum drying are used for probiotics and microorganism cultures that will be used as starters or biological additives in the fermentation. The same development is observed in the preservation methods, and studies with plant extracts and essential oils have been proposed and introduced, replacing chemical agents, such as propionate, within the clean-label bread formulations concept. Baking science is a growing research line incorporating innovative methods and biological additives.
ARTICLE | doi:10.20944/preprints202309.1792.v1
Subject: Biology And Life Sciences, Aquatic Science Keywords: phytoplankton; diversity; bioindicators; water quality indices; thermal impact; Pechorskoe Reservoir; European North
Online: 27 September 2023 (04:24:56 CEST)
Abstract: The species composition, the structure of phytoplankton communities, hydrochemical parameters in the artificial cooling reservoir of one of the largest thermal power plants (TPP) in the north-east of Europe in the Pechora River basin, Komi Republic were studied in the ice-open period in June and August. Altogether 81 species of algae and cyanobacteria was revealed with the last one predominated. Among cyanobacteria and algae (Bacillariophyta, Chlorophyta), indicators of water quality were found. Assessment of water quality according to the species composition and abundance of indicator phytoplankton species showed that the waters of the Pechorskoe Reservoir can be at-tributed to the III class of water quality, beta-mesosaprobic self-purification zone, with water quality of satisfactory purity, retaining the ability to self-purify. An increase in the diversity of plankton communities and high values of their abundance and biomass under the influence of the warming effect of TPP water outlet were noted and revealed its spatial distribution over the reservoir surface. The plankton of the northern reservoir under the influence of warm waters indicates a slight in-crease in the trophic status of the studied water body, which is also confirmed by the data of hydrochemical analysis. Ecological mapping for the chemical and biological variables as well as pollution indices over the water area of the reservoir was carried out. The results obtained can be used to make forecasts of changes in the phototrophic biota of small northern water bodies under the conditions of climate change.
REVIEW | doi:10.20944/preprints202309.1793.v1
Subject: Biology And Life Sciences, Neuroscience And Neurology Keywords: neurodegenerative disorders; blood-brain barrier; non-invasive delivery; device-related delivery; Alzheimer’s; Parkinson’s; ALS; Down Syndrome
Online: 27 September 2023 (04:19:06 CEST)
Device-mediated, non-invasive drug delivery across the blood-brain barrier (BBB) represents a significant advancement in treating neurological diseases. The BBB is a tightly packed layer of endothelial cells that shields the brain from harmful substances in the blood, allowing necessary nutrients to pass through. It is a highly selective barrier, which poses a challenge to delivering therapeutic agents into the brain. Several non-invasive techniques and devices have been proposed or investigated to enhance drug delivery across the BBB. This paper presents the current state of the art and case studies that address the pharmacology, technology, delivery systems, regulatory approval, ethical concerns, and future possibilities.
ARTICLE | doi:10.20944/preprints202309.1810.v1
Subject: Biology And Life Sciences, Insect Science Keywords: Glossina morsitans; Rabbits; Guinea pigs; Rodents; Squirrels; bloodfed; and attracted
Online: 26 September 2023 (16:12:13 CEST)
The success of any tsetse control programs depends on the knowledge of their behaviour. This study assessed the host choice and feeding behaviours of Glossina morsitans’ siblings whose parents were bloodfed on Rabbits, Guinea pigs, Rodents and Squirrels. Individual host was placed in a screen cage which allowed flies to enter through openings on each side. The groups of flies (20 per replicate) colour-marked differently basing on their parents’ bloodmeal hosts, were released from the centre of large semi-field cage. Released flies were aspirated after 24 hours, then, sorted basing on the location, feeding status and parents’ bloodmeal. A total of 213 flies (72.95% of the recovered) were attracted to the hosts. The number of flies attracted to different hosts varied significantly (χ²4= 33.685, p= 0.0001); Rodent (n=80, p=0.006), Rabbit (n=59, p=0.331), Guinea pig (n=49, p=0.057) and squirrel (n=25, p=0.005). The number of flies attracted to their parent’s blood meal source varied significantly (χ²12 = 56.476, p<0.001); rabbits (n= 35, 59.32%, p<0.001), rodent (n=25, 31.25%, p=0.043) and guinea pig (n= 19, 38.78%, p=0.45). But, only 39 flies (18.31% of total attracted) bloodfed on the hosts; Guinea pigs (n=10, 25.64%), Rodents (n=23, 58.97%), Rabbits (n=6, 15.38%) and Squirrels (n=0,0.0%). There was significant variation in number of flies that fed successively across hosts (χ²4=49.478, p<0.001). The findings from this study confirms the presence of the hosts’ differential attractiveness to flies but failed to explain observed behaviours with reference to genetic inheritance. Therefore, the study attracts the need for detailed investigation on the influence of bloodmeal sources on tsetsefly siblings’ behaviours across filial generations using small mammals.
REVIEW | doi:10.20944/preprints202309.1714.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: Cognitive Computing; Digital Imaging; Sound Vocalization Analysis; Animal Self-awareness; Behavioral Predictions; Micro-expressions; Animal Welfare Enhancement; Human-Animal-Computer Interactions
Online: 26 September 2023 (13:43:02 CEST)
In this paper, we critically examine the burgeoning role of advanced computational methodologies in deciphering the complex tapestry of farm animal behaviors and emotions. Leveraging digital imaging and artificial intelligence, we unearth nuanced behavioral patterns and micro-expressions, offering predictive insights into animal emotional states. Sound vocalization analysis, often overlooked, emerges as a pivotal tool, decoding intricate communicative nuances and emotional undertones. Cognitive tests, including mirror and bias assessments, challenge long-standing perceptions, revealing surprising depths of animal self-awareness and cognitive sophistication. However, the paper also underscores the imperative of integrating these tools with a profound understanding of animal psyche, ensuring technology serves as an enhancer, not a replacement, of traditional observational methods. This research not only highlights the transformative potential of cognitive computing in animal welfare but also calls for a judicious application, ensuring technology augments, not undermines, the intrinsic value of human-animal interactions and understanding.
ARTICLE | doi:10.20944/preprints202309.1757.v1
Subject: Biology And Life Sciences, Virology Keywords: Herpes B virus; zoonosis; LAMP; surveillance; cynomolgus macaque
Online: 26 September 2023 (13:39:13 CEST)
Herpes B virus (BV) is a zoonotic virus which can be transmitted from macaques to humans, which is often associated with high mortality rates. Because macaques often exhibit asymptomatic infections, individuals who come into contact with these animals face unexpected risks of BV infections. Serological test is widely performed to investigate BV infections. However, the assay’s sensitivity and specificity appeared to be inadequate, and it does not necessarily indicate ongoing viral shedding. Here, we developed LAMP and qPCR assays aiming to detect BVs with high sensitivity and specificity in various macaque species and validated them using oral swab samples collected from 97 wild cynomolgus macaques living in Thailand. Our LAMP and qPCR assays detected more than 50 and 10 copies of the target sequences per reaction, respectively. The LAMP assay could detect BV within 25 min, indicating its advantages for the rapid detection of BV. Collectively, our findings indicated that both assays developed in this study exhibit advantages and usefulness for BV surveillance and diagnosis of BV infections in macaques. Furthermore, for the first time, we determined the partial genome sequences of BVs detected in cynomolgus macaques in Thailand. Phylogenetic analysis revealed the species-specific evolution of BV within macaques.
REVIEW | doi:10.20944/preprints202309.1738.v1
Subject: Biology And Life Sciences, Aging Keywords: Sarcopenia; Zebrafish; Age-related muscle deficits; Aging; Biomarkers; Electrical Impedance Myography
Online: 26 September 2023 (13:38:16 CEST)
Abstract: Sarcopenia poses a significant challenge to public health and can severely impact the quality of life of aging populations. Despite extensive efforts to study muscle degeneration using traditional animal models, there is still a lack of effective diagnostic tools, precise biomarkers, and treatments for sarcopenia. Zebrafish models have emerged as powerful tools in biomedical research, providing unique insights into age-related muscle disorders like sarcopenia. The advantages of using zebrafish models include their rapid growth outside of the embryo, optical transparency during early developmental stages, high reproductive potential, ease of husbandry, compact size, and genetic tractability. By deepening our understanding of the molecular processes underlying sarcopenia, we may develop novel diagnostic tools and effective treatments that can improve the lives of aging individuals affected by this condition. This review aims to explore the unique advantages of zebrafish as a model for sarcopenia research, highlight recent breakthroughs, outline potential avenues for future investigations, and emphasize the distinctive contributions that zebrafish models offer. Our research endeavors to contribute significantly to addressing the urgent need for practical solutions to the impact of sarcopenia on aging populations, ultimately striving to enhance the quality of life for individuals affected by this condition.
ARTICLE | doi:10.20944/preprints202309.1776.v1
Subject: Biology And Life Sciences, Toxicology Keywords: lung cancer; dehydroepiandrosterone; proliferation; viability; migration; autophagy
Online: 26 September 2023 (11:59:48 CEST)
Lung cancer is a prevalent and aggressive neoplasm worldwide, contributing to significant mortality rates. Dehydroepiandrosterone (DHEA) constitutes the bulk of the steroid hormone in human plasma, has a robust antiproliferative effect, and induces cell death in various tumor cells. However, its role in lung cancer cells remains unexplored. This study aimed to investigate the influence of DHEA on the proliferation, viability, autophagy, and migration of several lung cancer cell lines, including A549, HCC827, and NCI-H2347. Cell proliferation was assessed through crystal violet staining; cell number and viability were evaluated using trypan blue staining; viability was confirmed by MTT reduction, a method that is also an indicator of metabolic activity; migration was assessed via a wound healing assay. Autophagy was evaluated using a specific kit, while cell death was determined by annexin-V-FITC/propidium iodide staining and caspase-3/7 activity assay. The results indicate that DHEA significantly reduced proliferation, cell number, metabolic activity, and migration in all examined lung tumor cells. These effects correlate with an increased autophagy induced by DHEA. No signs of apoptosis or necrosis were observed across the range of DHEA concentrations used. Although these findings are preliminary, they suggest that DHEA could hold promise as an alternative treatment option for various subtypes of lung cancer.
BRIEF REPORT | doi:10.20944/preprints202309.1754.v1
Online: 26 September 2023 (11:50:21 CEST)
Protein synthesis has been a very rich target for developing classes of drugs to control prokaryotic and eukaryotic pathogens. Despite the development of new drug formulations, treating human cutaneous and visceral Leishmaniasis still needs significant improvement due to considerable side effects and low adherence to the usual treatment regimen. In this work, we show that the di-substituted urea-derived compound I-17 is effective in inhibiting the promastigote forms and intracellular amastigotes of the Leishmania (L.) amazonensis and L. infantum species, in addition to exhibiting low macrophage cytotoxicity. We also show a potential immunomodulatory effect of I-17 in infected macrophages, which exhibited increased expression of inducible Nitric Oxide Synthase (NOS2) and Nitric Oxide (NO) production. Our data suggest that I-17 and new derivatives of this compound may be helpful in developing new drugs for treating leishmaniasis.
ARTICLE | doi:10.20944/preprints202309.1790.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: TAp73; PDAC; RAC1b; Biglycan; SMAD3; SMAD4; transforming growth factor-β; epithelial-mesenchymal transition; cell migration
Online: 26 September 2023 (11:48:58 CEST)
Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease due to early metastatic spread, late diagnosis and the lack of efficient therapies. A major driver of cancer progression and hurdle to successful treatment is the desmoplastic reaction of the tumor stroma, the formation of which is orchestrated by transforming growth factor (TGF)-β. Recent data from pancreatic cancer mouse models have shown that the TGF-β pathway is controlled by transcriptionally active p73 (TAp73) through secretion of biglycan (Bgn) via intermittent expression of the TGF-β signaling intermediates, Smad3 and Smad4. Genetic knockout of TP73, and, as a consequence, deficient induction of Smad3/Dpc4 and secretion of Bgn led to activation of TGF-β signaling through a (Smad-independent) ERK pathway, favoring epithelial-mesenchymal transition (EMT) and cell motility. Except for BGN, these functions of TAp73 have recently been shown to also operate in human PDAC cells and are reminiscent of what we previously observed for the small GTPase, RAC1b. This prompted us to hypothesize that TAp73 and RAC1b are part of the same tumor-suppressive pathway in human PDAC cells. The two objectives of this study, therefore, were to reveal i) if the regulatory interactions between TAp73 and Bgn previously discovered in murine PDAC-derived cells also operate in their human counterparts, and ii) if RAC1b collaborates with TAp73 in these tumor-suppressive activities in human PDAC cells. Using a variety of experimental approaches, including mutual rescue experiments, we were able to show that the previously proposed tumor-suppressive TAp73-Smad4-Bgn signaling also operates in human cells and that RAC1b is as an upstream activator of this pathway. Our findings highlight the complex role of TGF-β in pancreatic tumorigenesis and might have implications for therapeutic approaches targeting this growth factor for inhibition.
REVIEW | doi:10.20944/preprints202308.0706.v2
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Interferon; Innate; Adaptive; Genetic; Molecular
Online: 26 September 2023 (11:35:53 CEST)
Interferons were the original prototype cytokine system discovered in 20th-century research. As the name implies, they were originally thought to be synthesised and secreted between cells. Thanks to technological advances, the processes involved in protein secretion can be explained comparatively more clearly at both the genetic and biochemical levels. The discovery of interferon (IFN) occurred when genetic research was still in its infancy. Franklin and Wilkins discovered the structure and function of deoxyribonucleic acid (DNA) at the same time as Crick and Watson; however, Isaacs and Lindemann, two scientists, described the first IFN in 1957. Mutations can be caused by inherent genetic protein synthesis and during infection as well as within IFN regulation pathways affecting cell proliferation. This remains central to host cell IFN synthesis and effects through IFN protein receptor subunits defined by 6 protein domains. Type II IFN is key to immune cell function secreted by a variety of immune cells, mainly natural killer (NK) as well as T cells. Single–stranded and/or double–stranded RNA/DNA viruses, as well as bacterial infections (e.g., Escherichia coli) and fungal infections (e.g., Aspergillus), also affect IFN regulation. Pathogenic proteins utilise intra/extracellular proteins that sense foreign antigens like Toll–like Receptors (TLRs), affected by mutations within the human cellular IFN transduction pathways. Since the discovery of the third IFN type in 2003, when immune cell phenotypes were further characterised, questions remain about the immunological mechanisms contributing to the regulation of the innate and adaptive host immune system. Alterations in the synthesis of type I/II/III host IFNs can differentially and beneficially alter homeostatic cellular pathways in pathological disease, with type I IFN being synthesised in cancer as well as by homeostatic cells. Therefore, considered here are the overall IFN molecular, cell regulatory mechanisms in the context of immune cell research developments.
ARTICLE | doi:10.20944/preprints202309.1758.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: membrane pump theory; membrane potential; cell shape; biophysics; biology
Online: 26 September 2023 (11:32:12 CEST)
Membrane potential generation and maintenance are fundamental parts of membrane theory. However, since its inception, theories and assumptions have remained simple, whereas the amount of knowledge available has only grown. Unfortunately, this simplification and reductionism lead to its loss of validity on both microscopic and macroscopic scales. This article shows that assumptions made outside the context of factual reality lead to contradictions that cannot be compared with science. Membrane theory must be thoroughly revised, taking into account all forgotten and unanswered assumptions and questions to conform to facts and science to maintain credibility.
ARTICLE | doi:10.20944/preprints202309.1783.v1
Subject: Biology And Life Sciences, Cell And Developmental Biology Keywords: human mesenchymal stem cells; mesenchymal to epithelial transition (MET); epithelial to mesenchymal transition (EMT); HAND2; HAND2-AS1; senescence; stem cell plasticity
Online: 26 September 2023 (10:46:24 CEST)
We have previously developed several successful decellularization strategies yielding porcine cardiac extracellular matrices (pcECMs), which exhibit tissue-specific bioactivity and bioinductive capacity when cultured with various pluri- and multipotent stem cells. Here, we studied the tissue-specific effects of the pcECM on seeded human mesenchymal stem cells (hMSCs) phenotype using reverse transcribed quantitative polymerase chain reaction (RT-qPCR) arrays for cardio-vascular related genes. We further corroborated interesting findings at the protein level (flow cytometry and immunological stains) as well as bioinformatically using several mRNA sequencing and protein databases of normal and pathologic adult tissue expression, as well as during human embryonic organogenesis. We discovered that upon seeding of human mesenchymal stem cells (hMSCs) on the pcECM they displayed partial MET toward endothelial phenotypes (CD31+) and morphologies, which were preceded by an early spike (~day 3 onward after seeding) in HAND2 expression at both the mRNA and protein levels compared to plate controls. CRISPR-Cas9 knockout (KO) of HAND2 and its associated antisense long non-coding RNA (HAND2-AS1) regulatory region resulted in proliferation arrest, hypertrophy, and senescent-like morphology. Bioinformatic analyses revealed that HAND2 and HAND2-AS1 are highly correlated in expression, are expressed in many different tissue types albeit at distinct yet tightly regulated expression levels. Deviation (down or up regulation) from these basal tissue expression levels are associated with a long list of pathologies. We thus suggest that HAND2 expression levels may finetune cell plasticity possibly affecting senescence and mesenchymal-to-epithelial transition states, through yet unknown mechanisms. Targeting this pathway may represent a promising new therapeutic approach for a wide range of diseases, including cancer, degenerative disorders, and aging. Nevertheless, further investigations are required to better understand the molecular players involved, potential inducers and inhibitors of this pathway, and eventually potential therapeutic applications.
ARTICLE | doi:10.20944/preprints202309.1782.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: Schistosomiasis; Mass Drug Distribution; Sentinel sites, Mali
Online: 26 September 2023 (10:12:34 CEST)
Background – Mali was one of the first countries in sub-Saharan Africa to initiate a National Schistosomiasis Control Programme (NSCP) in 1982. The WHO's 2021-30 roadmap sets out criteria for eliminating and controlling schistosomiasis as a public health problem. Our study aimed to assess the impact of annual Mass Drug Distribution (MDD) with praziquantel (PZQ) among school-age children in the sentinel sites (SS) of the NSCP. Methods –The study took place at twelve SS in Kayes and Koulikoro regions. Two-round observational cross-sectional studies were carried out in December 2014-2015 and in April 2018 after four to five years of annually MDD. Overall, 2442 schoolchildren aged 7 to 14 were successfully examined. The urine filtration and Kato-Katz method were used for determining Schistosoma haematobium and S. mansoni eggs, respectively. Results –Of the twelve SS treated from 2014-2015, one has achieved the criterion of elimination of S. haematobium as a public health problem (prevalence of heavy intensity infection PHI < 1%) (ie, ≥50 S. haematobium eggs per 10 mL of urine or ≥400 S. mansoni eggs per g of stool), four met the morbidity control criterion (PHI< 5%) while two sites remained confined below the morbidity control criterion (PHI>5%). Five SS had no heavy intensity infection. The prevalence of S. mansoni was less than 1%. Conclusion –The impact evaluation of MDD with praziquantel in the SS of NSCP highlights that MDD has significantly reduced the PHI of schistosomiasis. However, the high prevalence of schistosomiasis or its increase in some sites requires in-depth studies.
REVIEW | doi:10.20944/preprints202309.1766.v1
Subject: Biology And Life Sciences, Animal Science, Veterinary Science And Zoology Keywords: animal welfare; photoreceptor; light; circadian; laboratory animal
Online: 26 September 2023 (08:41:01 CEST)
Light enables vision and exerts widespread effects on physiology and behaviour, including regulating circadian rhythms, sleep, hormone synthesis, affective state, and cognitive processes. Appropriate lighting in animal facilities may support welfare and ensure that animals enter experiments in a controlled physiological and behavioural state. Proper consideration of light during experimentation - both when it is explicitly employed as an independent variable and as a general feature of the environment - has the potential to provide more informative experimental designs and more reliable outcomes, contributing to Reduction and Refinement, helping to ensure more ethical animal use, and improving data quality. As such, it is unfortunate that ambient light for animals is typically quantified in units (lux) designed for human observers. We report the consensus views of an expert working group, with expertise spanning mammalian photobiology, neurobiology and animal husbandry and welfare, convened in February 2023 to agree upon metrics for light appropriate for non-human mammals and their application to improve animal welfare and the quality of animal research. We conclude that species-specific versions of the recently standardised a-opic metrology represent the best available approach to quantifying light. We provide methods for measuring these quantities; practical guidance for their implementation in husbandry and experimentation; and quantitative guidance on appropriate light exposure for laboratory mammals.