Manual material handling tasks in industry cause work-related musculoskeletal disorders. Exoskeletons are being introduced to reduce the risk of musculoskeletal injuries. This study investigated the effect of using a passive lumbar exoskeleton in terms of moderate ergonomic risk. Eight participants were monitored by electromyogram (EMG) and motion capture (MoCap) while performing tasks with and without the lumbar exoskeleton. The results showed a significant reduction in the root mean square (VRMS) for all muscles tracked: erector spinae (8%), semitendinosus (14%), gluteus (5%), and quadriceps (10.2%). The classic fatigue parameters showed a significant reduction in the case of the semitendinosus: 1.7% zero-crossing rate, 0.9% mean frequency, and 1.12% median frequency. In addition, the logarithm of the normalized Dimitrov’s index showed reductions of 11.5, 8, and 14% in erector spinae, semitendinosus, and gluteus, respectively. The calculation of range of motion in the relevant joints demonstrated significant differences, but in almost all cases, the differences were smaller than 10. The findings of the study indicate that the passive exoskeleton reduces muscle activity and introduces a minor change of strategies for motion. Thus, EMG and MoCap appear to be appropriate measurements for designing an exoskeleton assessment procedure.

Cost-effective and functionalized scaffolds are in high demand for stem-cell-based regenerative medicine to treat refractory bone defects in craniofacial abnormalities and injuries. One potential strategy is to utilize pharmacological and cost-effective plant polyphenols and biocompatible proteins, such as gelatin. Nevertheless, the use of chemically modified proteins with plant polyphenols in this strategy has not been standardized. Here, we demonstrated that gelatin chemically modified with epigallocatechin gallate (EGCG), the major catechin isolated from green tea, can be a useful material for dedifferentiated fat cells and adipose-derived stem cells and can induce bone regeneration in a rat congenial cleft-jaw model in vivo. Vacuum-heated gelatin sponge modified with EGCG (vhEGCG-GS) induced superior osteogenesis from these two cell types compared with vacuum-heated gelatin sponge (vhGS). The EGCG-modification converted the water wettability of vhGS to a hydrophilic property (contact angle: 110° to 3.8°) and the zeta potential to a negative surface charge; the modification enhanced the cell adhesion property and promoted calcium phosphate precipitation. These results suggest that the EGCG-modification with chemical synthesis can be a useful platform to modify the physicochemical property of gelatin. This alteration is likely to provide a preferable microenvironment for multipotent progenitor cells, inducing superior bone formation in vivo.

Numerous researchers and institutions have been developing in ovo sexing technologies to improve animal welfare by identifying male embryos in an early embryonic stage and disposing of them before pain perception. This review gives a complete overview of the technological approaches reported in papers and patents by performing a thorough search using Web of Science and Patstat databases for papers and patents, respectively. Based on a total of 45 papers and 100 patent families up until 2021 reported worldwide, 11 technology categories were defined: six non-optical and five optical techniques. Every category was described for its characteristics while assessing its potential for application. Next, the dynamics of the publications of in ovo sexing techniques in both paper and patent fields were described through growth curves, and the interest or actual status was visualized using the number of paper citations and the actual legal status of the patents. When comparing the reported technologies in papers to those in patents, scientific gaps were observed, as some of the patented technologies were not reported in the scientific literature, e.g., ion mobility and mass spectrometry approaches. Generally, more diverse approaches in all categories were found in patents, although they do require more scientific evidence through papers or industrial adoption to prove their robustness. Moreover, although there is a recent trend for non-invasive techniques, invasive methods like analyzing DNA through PCR or hormones through immunosensing are still being reported (and might continue to be) in papers and patents. It was also observed that none of the technologies complies with all the industry requirements. Thus, more research and harmony between consumers, industry, and governments is necessary.

Cryopreservation has been used extensively for cattle in Bangladesh, albeit no study was conducted on the cryopreservation techniques of buffalo. This study compares two freezing methods and the effects of diluters on the semen quality of buffalo. In the first freezing protocol, semen was frozen in two-step: from 37 °C to 5 °C for 30 minutes in a BLRI-developed equilibration chamber and from 5 oC to -120 oC in a Styrofoam box using liquid nitrogen vapor from different distances (0.5, 1.5, 1.6, 2 and 3 inches). At the same time semen was frozen in a programmable freezer in three steps. The semen samples were then evaluated for motility and morphological quality by CASA. In another experiment, the efficacy of one locally developed diluter-Tris-fructose-egg yolk-based (TFE) and three commercial diluters (Andromed, Triladyl and Steridyl) were evaluated. The highest number of motile sperms (62.67±1.12; P< 0.01) and progressive motility (38.97±1.10; P < 0.001) was observed at 1.6 inches above liquid nitrogen. There was no significant difference in overall motility, progressive and slow motility between semen cryopreserved in the cheap nitrogen vapor technique (57.49±5.67, 38.70± 4.04 and 3.83± 0.63, respectively) and expensive automated technique (65.94±4.65, 45.54 ± 3.64 and 2.43± 0.36, respectively). The highest recovery rate and conception rate were observed in semen diluted with TFE (82.4% and 80%, respectively). Hence, the cryopreservation technique using nitrogen vapor and TFE diluent is cost-effective and suitable for freezing buffalo semen that would produce superior semen for artificial insemination.

Cone snails (genus Conus) are venomous marine snails that inject prey with a lethal cocktail of conotoxins, small, secreted, cysteine-rich peptides. Given the diversity and often high affinity for their molecular targets, consisting of ion channels, receptors or transporters, many conotoxins have become invaluable pharmacological probes, drug leads and therapeutics. Transcriptome sequencing of Conus venom glands followed by de novo assembly and homology-based toxin identification and annotation is currently the state-of-the-art for discovery of new conotoxins. However, homology-based search techniques, by definition, can only detect novel toxins that are homologous to previously reported conotoxins. To overcome these obstacles for discovery we have created ConusPipe, a machine learning tool that utilizes prominent chemical characters of conotoxins to predict whether a certain transcript in a Conus transcriptome, which has no otherwise detectable homologs in current reference databases, is a putative conotoxin. By using ConusPipe on RNASeq data of 10 species, we report 5,230 new putative conotoxin transcripts that have no homologues in current reference databases. 893 of these were identified by at least 3 out of 4 models used. These data significantly expand current publicly available conotoxin datasets and our approach provides a new computational avenue for the discovery of novel toxin families.

In Cameroon, alcoholic beverages remain the main consumed drink. In the Northern regions, indigenous sorghum beers are popular and widely consumed in an actively fermenting state by the people. In this study, some physicochemical parameters of the alcoholic sorghum beverages and correlations between them were evaluated during fermentation for 10 days. The indigenous white and red beers were produced at the laboratory scale assisted by experimented producers and some parameters (pH, total acidity, alcohol, sugars, density, total solids, temperature, and conductivity) were measured on the wort and fermented beverages. The pH decreases from 3.2 to 2.4 and 3.11 to 2.41; total acidity increases from 1.07 to 5.1 g/l and 0.5 to 4.6 g/l; alcohol enhances from 0 to 9.5% and 0 to 6.8% (v/v); total solids drop from 13.6 to 5°P and 12.2 to 3.3°P were observed respectively in the white and red sorghum beers. The multivariate analysis showed a good correlation between consumption of sugar, the decrease of total solids and density with the decrease in pH. It was also shown that, a perfect link exist between the production of alcohol and organic acids. The hierarchical analysis showed that indigenous red beer samples fermented for 1 and 2 days and those fermented for 4 to 10 days are related and could be separate in two distinct groups, whereas white turbid beer samples are separated in three different groups, those fermented for 1 to 4 days, those 5 to 6 days and those 7 to 10 days. The results obtained could serve as a guide to better understand the fermentation process of the indigenous alcoholic sorghum-based beverages.

Physiologically Based Pharmacokinetic Models (PBPK) are mechanistical tools generally employed in the pharmaceutical industry and environmental health risk assessment. These models are recognised by regulatory authorities for predicting organ concentration-time profile, pharmacokinetic and daily intake dose of xenobiotics. Extension of PBPK models to capture sensitive populations like pediatric, geriatric, pregnant females, fetus etc. and diseased population like renal impairment, liver cirrhosis etc. is a must. However, the current modelling practice and existing models are not mature enough to confidently predict the risk in these populations. A multidisciplinary collaboration between clinicians, experimental and modeler scientist is vital to improve the physiology, and calculation of biochemical parameters for integrating the knowledge and refining existing PBPK models. Specific PBPK covering compartments like cerebrospinal fluid, and hippocampus are required to gain mechanistic understanding about xenobiotic disposition in these sub-parts. The PBPK model assists in building quantitative adverse outcome pathways (qAOPs) for several endpoints like developmental neurotoxicity (DNT), hepatotoxicity and cardiotoxicity. Machine learning algorithms can predict physicochemical parameters required to develop in-silico models where experimental data is unavailable. Integrating machine learning with PBPK carries the potential to revolutionize the field of drug discovery and development and environmental risk. Overall, this review tried to summarize the recent developments in the in-silico models, building qAOPs, use of machine learning for improving existing models along with a regulatory perspective. This review can act as a guide for toxicologists who wish to build their careers in kinetic modeling.

This paper reports for the first time on the production of probiotic alcoholic and non-alcoholic rosé wines with enhanced health benefits made with Saccharomyces cerevisiae var. boulardii probiotic yeast. The alcohol, sugar, volatile acidity lactic and malic acid contents were assessed for S. cerevisiae var. boulardii before and after fermentation and distillation and compared with a conventional Saccharomyces cerevisiae (ex-bayanus) yeast. The free amino nitrogen and gluconic acid concentrations in the musts were determined. Yeast viability was evaluated after fermentation and distillation as a function of time (0, 15 days, 3 months and 6 months) both at room temperature (25±0.5ºC) and refrigerator temperature (4±0.5ºC). The results obtained showed that the probiotic rosé wine produced with S. cerevisiae var. boulardii possesses the typical values and sensory attributes of other commercial wines produced with S. cerevisiae (ex-bayanus). The probiotic S. cerevisiae var. boulardii yeast survives the high alcohol content produced during fermentation and vacuum distillation. The study also showed that this probiotic rosé wine stored either at room temperature or in a refrigerator keeps its probiotic viability for at least six months, which makes it a promising for large-scale production, in which long storage times are required by both producers and consumers.

Rapid progress in gene editing based on clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) has revolutionized the study of gene and genome function and genetic disease correction. While numerous genetically modified cellular and animal models have been created to understand biological processes, the clinical application of CRISPR/Cas tools has been impeded by off-targeting and delivery problems. It is generally accepted that the delivery of CRISPR in the form of a ribonucleoprotein complex (RNP) substantially reduces the time of DNA exposure to the effector nuclease, minimizing off-target effects and facilitating clinical usage. This review focuses on CRISPR/Cas RNP delivery with retro/lentiviral particles and exosomes, whose parallel production by cells transfected with viral vectors is underestimated. We critically evaluate specific mechanisms of extracellular particle formation and loading with CRISPR/Cas for each system. Additionally, the details of Cas-nanoparticle entry and uncoating, previously unappreciated in the context of gene editing efficiency, are discussed. Based on existing knowledge about the consequences of intervention in retroviral assembly, entry, or exosome formation, we outline the potential problems with CRISPR/Cas delivery using extracellular nanoparticles and ways to address them.

: Microbial fuel cell (MFCs) devices utilise the metabolic processes of bacteria to generate electricity from various organic substrates. Due to the enormous amount of energy that organic waste produces, scientists are highly interested in advanced MFCs. MFCs serve as a multidisciplinary research platform at the engineering and natural sciences intersection. MFCs have various uses besides being used as energy sources, such as sensing capabilities. Despite showing considerable promise, only a few marine sediment MFCs have been deployed in real-world applications to supply current for low-power devices. It is now required to maintain track of the work being done by research groups worldwide and regularly compile significant discoveries due to the rising quantity of research publications. Review papers are a traditional beginning point for a literature review for new scholars. This review is a fast reckoner that directs readers to pertinent reviews and research publications detailing significant advances in microbial fuel cell research during the previous two decades. An overview of key advances in MFC research over the past two decades is provided as a quick reference in this review article. In addition, the report identifies research gaps that, if filled, could bring this technology closer to real-world use.

Heat shock transcription factors (HSFs) participate in regulating many environmental stress responses and biological processes in plants. Maize (Zea mays L.) is a major cash crop that is grown worldwide. However, the growth and yield of maize are affected by several adverse environmental inputs. Therefore, investigating the factors that regulate maize growth and development and resistance to abiotic stress is an essential task for developing stress-resilient maize varieties. Thus, a comprehensive genome-wide identification analysis was performed to identify HSFs in the maize genome. The current study identified 25 ZmHSFs, randomly distributed throughout the maize genome. Phylogenetic analysis revealed that ZmHSFs are divided into three classes and 13 sub-classes. Gene structure and protein motif analysis supported the results obtained through the phylogenetic analysis. Domain analysis showed the DNA-binding domain to be the most conserved region of ZmHSFs. Segmental duplication is shown to be responsible for the expansion of ZmHSFs. Most of the ZmHSFs are localized inside the nucleus, and the ZmHSFs which belong to the same group show similar physio-chemical properties. The 3D structures revealed comparable conserved ZmHSFs protein structures. RNA-seq analysis revealed a major role of class A HSFs including, ZmHSFA-1a and ZmHSFA-2a in all the maize growth stages, i.e., seed, vegetative, and reproductive development. Furthermore, ZmHSFs displayed an obvious spatiotemporal expression. Under abiotic stress conditions (heat, drought, cold, UV, and salinity), members of class A and B ZmHSFs are induced. Gene ontology (GO) annotation analysis indicated a major role of ZmHSFs in resistance to environmental stress and regulation of primary metabolism. Further, the protein-protein interaction analysis showed that ZmHSFs interact with several molecular chaperons and major stress-responsive proteins. To summarize, this study provides novel insights for functional studies on the ZmHSFs in maize breeding programs.

Fish oil rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), vitamin D₃ and natural astaxanthin sustainably extracted from anchovy fillet leftovers using biobased limonene only, can be conveniently stabilized by adsorption on periodic mesoporous silicas. The simplicity of the process, the high load of entrapped fish oil, and the biocompatible nature of silica support numerous forthcoming applications of this new class of “Omeg@Silica” materials.

A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules such as nucleotides possess vital role in cell metabolism, energy economy, and signaling. Thus the monitoring of nucleotide concentration and nucleotide related enzymatic reactions is of high importance. Two component lanthanide complex formed from Tb(III) ion carrier and light harvesting antenna, readily distinguishes nucleotides containing different number of phosphates and enable direct detection of enzymatic reactions converting nucleotide triphosphate (NTP) to nucleotide di/monophosphate or the opposite. Developed sensor enables the detection of enzymatic activity with a low nanomolar sensitivity, as highlighted with K-Ras and apyrase enzymes in there hydrolysis assays performed in high throughput screening compatible 384-well plate format.

Functional foods are essential food products that possess health-promoting properties for the treatment of infectious diseases. In addition, they provide energy and nutrients, which are required for growth and survival. They occur as prebiotics or dietary supplements, including oligosaccharides, processed foods, and herbal products. However, oligosaccharides are more efficiently recognized and utilized, as they play a fundamental role as functional ingredients with great potential to improve health in comparison to other dietary supplements. They are low molecular weight carbohydrates with a low degree of polymerization. They occur as fructooligosaccharide (FOS), inulooligosaccharadie (IOS), and xylooligosaccahride (XOS), depending on their monosaccharide units. Oligosaccharides are produced by acid or chemical hydrolysis. However, this technique is liable to several drawbacks, including inulin precipitation, high processing temperature, low yields, high production costs, etc. As a consequence, the application of microbial enzymes for oligosaccharide production is recognized as a promising strategy. Microbial enzymatic production of FOS and IOS occurs by submerged or solid-state fermentation in the presence of suitable substrates (sucrose, inulin) and catalyzed by fructosyltransferases and inulinases. Incorporation of FOS and IOS enriches the rheological and physiological characteristics of foods. They are used as low cariogenic sugar substitutes, suitable for diabetics, and as prebiotics, probiotics & nutraceutical compounds. In addition, these oligosaccharides are employed as anticancer & antioxidant agents and aid in mineral absorption, lipid metabolism, immune regulation etc. This review, therefore, focuses on the occurrence, physico-chemical characteristics, and microbial enzymatic synthesis of FOS and IOS from coprophilous fungi. In addition, the potential health benefits of these oligosaccharides were discussed in detail.

The rapid development of algorithms for skeleton detection with relatively inexpensive contactless systems and cameras opens the possibility of virtual exercise therapy for patients with different complications. However, evaluation and confirmation of posture classifications is still needed. The purpose of this study was therefore to find the most accurate algorithm for automatic classification of human exercise movement. A Kinect V2 with 25 joints identification was used to record movements for data analysis. A total of 10 subjects volunteered for this study. Four algorithms were tested for the classification of different postures in Matlab. These were based on: total error of vector lengths, total error of angles, multiplication of these two parameters and simultaneous analysis of the first and second parameters. A base of 13 exercises was then created to test the recognition of postures by the algorithm, and to analyse subject performance. The best results for posture classification was shown by the second algorithm with an accuracy of 94.9%. The average correctness of exercises among the 10 participants was 94.2% (SD1.8%). The algorithms tested in this study therefore proved to be effective and could potentially form the basis for developing a system for remote monitoring of rehabilitation involving exercise.

RNA epigenetics is perhaps the most recent aspect of interest for translational epigeneticists. RNA modifications create such an extensive network of epigenetically driven combination whose role in physiology and pathophysiology is still far from being elucidated. Not surprisingly, some of the players determining changes into RNA structure are in common with those involved in DNA and chromatin structure regulation, while other molecules seem very specific to RNA. It is envisaged, then, that new small molecules, acting selectively on RNA epigenetic changes, will be reported soon, opening new therapeutic interventions based on the correction of the RNA epigenetic landscape. In this review, we shall summarize some aspects of RNA epigenetics limited to those in which the potential clinical translatability to cardiovascular disease is emerging.

Natural astaxanthin is a precious substance obtained from some organisms such as microalgae. This plant has many benefits for humans, so research into its cost-effective and economical production has recently increased. For this purpose, some methods such as the use of different culture media, gene engineering, different stresses, nanoparticles, bio acids, and phytohormones are important. Accordingly, this review study was conducted to demonstrate the effect of the factors mentioned above for the high production of astaxanthin in microalgae, especially Haematococcus pluvialis (H.p).

Non-viral gene carriers because of their limited side effects, biocompatibility, simplicity and taking the advantages of electrostatic interactions have shown noticeable potential in gene delivery. The low transfection rate of non-viral vectors under physiological conditions is a significant issue. Here, the aim of this study was to investigate the efficacy of hydrophilic and hydrophobic groups on gene carriers such as two synthesized amphiphilic polymer of dextran-stearic acid-spermine (DSASP) with verified lipid and amine conjugations that associated with Fe3O4 superparamagnetic nanoparticles to promote the target delivery and decrease the transfection time using static magnetic field. Our findings illustrate that magnetic nanoparticles are spherical with positive surface charges and superparamagnetic behaviors. The DSASP–pDNA/MNPs offered a strong pDNA condensation, protection against DNase degradation, significant cell viability in HEK 293T cells and. Although conjugations of spermine play a critical role in transfection efficiency, amphiphilic polymer with more derivatives of stearic acid showed better transfection yields. Therefore, DSASP amphiphilic magnetic carriers offer new insights for gene delivery due to the amine contents and ameliorate the uptake of complexes via cell membrane based on its hydrophilic surface.

Schistosomiasis, a disease historically associated with poverty, lack of sanitation and social inequalities, is a chronic, debilitating parasitic infection, affecting hundreds of millions of people in endemic countries. Although schistosomiasis control approach has shown that chemotherapy is capable of reducing morbidity in humans, rapid re-infection is a reminder that the impact of drug treatment on transmission control or elimination initiatives is marginal. In addition, and regardless of more than two decades of well-executed control activities based on large-scale chemotherapy, the disease is expanding in many areas including Brazil. The development of the Sm14/GLA-SE schistosomiasis vaccine is an emblematic open knowledge innovation that has successfully completed Phase I and Phase IIa clinical trials, with Phase II/III trials underway in the African continent and to be followed in Brazil. Discovery and experimental phases were long term achievements leading to a robust collection of data that are strongly supporting the presently ongoing Clinical Phase. This paper reviews the development of the Sm14 vaccine formulated with GLA-SE (Glucopyranosyl Lipid A), from the earlier experimental developments to clinical trials including the recent status of Phase II studies.

Red plum apricot is the best apricot over the world. Since having been introduced into Guyuan county in 1987 from Shaanxi province, red plum apricot grows well and then has been selected as good varieties to popularize in semirid loess hilly regions because the color of red plum apricot is beautiful, aroma fragrant , and taste is the mix of sour and sweet and nutrition is rich. Since 1995, the yield, benefits and planting area of red plum apricot increase doubly, and the distribution range of red plum apricot increased from Guyuan to the whole Ningxia, and then to Gansu province and so on in the water-limited regions, China. However, there are still some problems in the sustainable produce of red plum apricot. In order to promote the development of red plum apricot and meet people's increasing demand, we should do intensive study of the relationship between red plum apricot and environment and the comparison of possible methods to reduce the bad influence of drought, low temperature and frost on quality and production of red plum apricot, and determine the soil water resources use limit by red plum apricot and the suitable leaf amount of red plum apricot when the planting density equal soil water carrying capacity for red plum apricot and select the best method to reduce or evade the bad influence of soil drought, low temperature, frost and soil drought on red plum apricot to ensure the sustainable produce of red plum apricot in Sustainable Produce of red plum apricot in water-limited regions.

The peanut (Arachis hypogaea L.) is recognized as one of the most important legume crops globally for its use in human food; it is widely distributed and cultivated in tropical and subtropical regions. The purpose of this study was to evaluate the cryopreservation of five peanut varieties conserved in the INIAP Germplasm Bank, testing cryopreservation methods, evaluating the germination percentage of whole seeds and embryonic shoots. Subsequently, two quantitative variables, shoot length and root, were evaluated. The average germination percentage of varieties and treatments was higher when embryonic axes were isolated with 99.31% than 86.06% seeds. The best germination percentage of the five varieties for seeds and embryonic shoots was obtained by the Peruvian variety with 88.13% and 92.50%. The best treatments by variety for the germination of whole seeds and embryonic axes were obtained by the treatment (desiccation and NL) for whole seeds (GS2) with 95.42% and embryonic axes with 92.83%. Ageing and cryopreservation treatments positively affected germination and seedling vigor in whole seeds and embryonic axes. The two quantitative variables, shoot and root length showed variability between the five varieties; significant differences were observed between the four treatments evaluated for whole seeds and embryonic axes. The three treatments for whole seeds (GS1, GS2 GS3) and the non-cryopreserved control treatment (GSC), as well as the treatments for embryonic axes (GEA1, GEA2 GEA3) and the non-cryopreserved control treatment (GEAC), obtained good survival. They germinate whole seeds and embryonic axes with sprout development (aerial part) and root formation. With the most effective treatments for whole seeds (GS2) and embryonic axes (GEA2), the cryopreservation of the national peanut collection of the INIAP Germplasm Bank could be started.

The primary screening of hybridoma cells is a time-critical and laborious step during the development of monoclonal antibodies. Often critical errors occur in this phase, which supports the notion that the generation of monoclonal antibodies with hybridoma technology is difficult to control and hence a risky venture. We think that it is crucial to improve the screening process to eliminate most of the immanent deficits of the conventional approach. With this new microarray-based procedure, several advances could be achieved: Selectivity for excellent binders, high throughput, reproducible signals, avoidance of misleading avidity (multivalency) effects, and simultaneous performance of competition experiments. The latter can directly be used to select clones of desired cross-reactivity properties. In this paper, a model system with two excellent clones against carbamazepine, two weak clones and blank supernatant has been designed to examine the effectiveness of the new system. The excellent clones could be detected largely independent of the IgG concentration, which is unknown during the clone screening since the determination and subsequent adjustment of the antibody concentration is not possible in most cases. Furthermore, in this approach, the enrichment, isolation, and purification of IgG for characterization is not necessary. Raw cell culture supernatant can be used directly, even when fetal calf serum (FCS) or other complex media had been used. In addition, an improved method for the oriented antibody-immobilization on epoxy-silanized slides is presented. Based on the results of this model system, we conclude that this approach should be preferable to most other protocols leading to many of false positives, causing expensive and lengthy confirmation steps to weed out the poor clones.

Within phototherapy, a grand challenge in clinical cancer treatments is to develop a simple, cost-effective, and biocompatible approach to treat this disease using ultra-low doses of light. Carbon-based materials (CBM), such as graphene oxide (GO), reduced GO (r-GO), graphene quantum dots (GQDs), and carbon dots (C-DOTs), are rapidly emerging as a new class of thera-peutic materials against cancer. This mini-review summarizes the progress in lasts years re-garding the applications of CBM in photodynamic (PDT) and photothermal (PTT) therapies for tumor destruction. The current understanding of the performance of modified CBM, hybrids and composites, is also addressed. This approach seeks to achieve an enhanced healing action by im-proving and modulating the properties of CBM to treat various types of cancer. Metal oxides, organic molecules, biopolymers, therapeutic drugs, among others, have been combined with CBM to treat cancer by PDT, PTT, or synergistic therapies.

This study examines the matrix metalloproteinase (MMP) activity in the healing of liver injuries treated with biological adhesives Tachosil^® and GelitaSpon^® and the elastic cyanoacrylate Adhflex^®. Hepatic lesions were induced in male rats using a Stiefel biopsy punch. Healing was assessed 2, 6, and 18 days after injury by quantifying tissue levels of MMP1, 2, 8, 9, and 13. Histopathological repair was evaluated using hematoxylin-eosin, Masson’s trichrome, and Periodic Acid Schiff (PAS) staining and immunohistochemical markers CD31 and CD68. The sealants contributed to complete healing. Histopathology and MMP findings indicate that Adhflex^® has slower degradation and a strong inflammatory reaction at the onset of healing. Early on, all MMPs showed higher levels in Adhflex^® and Tachosil^®-treated animals, and MMP2 and MMP9 expressions were significantly higher in the Adhflex-treated group at 18 days post-injury (T3). The Adhflex^® group had significantly higher MMP8 and MPP13 levels than other treated groups and showed a sustained overexpression of all MMPs, even in the latest healing stages. Notably, the overexpression did not negatively influence the histological healing process. All hepatic trauma injuries should be treated as emergencies, and any easy-to-use and rapid sealant like Adhflex^® could be considered as an option for treating liver trauma.

Next generation sequencing and metabolomics have become very cost and work efficient and are integrated into an ever growing number of life science research projects. Typically, well established software pipelines provide quantitative data informing about gene expression or concentrations of metabolites from the raw data. This data needs to be visualized and further analyzed in order to support scientific hypothesis building and identification of underlying biological patterns. Some tools exist, but require installation or manual programming. We developed “Gene Expression Plotter” (GXP), an RNA-Seq and Metabolomics data visualization and analysis tool entirely running in the user’s web browsers, thus not needing any custom installation, manual programming or upload of confidential data to third party servers. GXP enables the user to generate interactive plots, visually summarize genetic or metabolic responses in scientific sketches (Mapman), carry out cluster and principal component analysis, and conduct overrepresentation analyses. GXP can be used to publish research data along with interactive plots and results of analyses carried out with it. GXP is freely available on GitHub: https://github.com/usadellab/GeneExpressionPlots

Over the past decade, the One Strain Many Compounds (OSMAC) approach has been established for silent gene cluster activation and elicitation of secondary metabolite production, but so far the full secondary metabolome of a biosynthetically promising bacterium has not been elucidated yet. Here, we investigate the ability of seven categories of OSMAC conditions to elicit new mass features from bacterial strains with little literature coverage but high biosynthetic potential. The strains B. amyloliquefaciens DSM7, C. coralloides DSM2259, P. fallax HKI727, R. jostii DSM44719 and S. griseochromogenes DSM40499 were selected after genome mining with antiSMASH. After cultivation under OSMAC conditions, the generated extracts were subjected to LC/MS and MZmine analysis to determine new mass features, expressed gene clusters and evaluate the tested culture conditions. 4 predicted compounds, bacillibactin, desferrioxamine B, myxochelin A and surfactin, were identified and up to 147 new mass features were detected in the generated extracts, which greatly surpasses the number of predicted gene clusters. Among the new mass features are bioactive compounds which were so far unreported for the strains such as cyclo-(Tyr-Pro) from DSM7 and nocardamin from DSM2259. Furthermore, the tested culture conditions were evaluated regarding their suitability for the generation of new mass features from the selected strains and promising new starting points for further screenings are postulated. Especially culture conditions with little prior literature coverage are responsible for the activation of secondary metabolite production.

The α(1,2)fucosyltransferase (Se enzyme) encoded by FUT2 is involved in the secretor status of ABH(O) blood group antigens. The sedel2 allele is one of the non-functional FUT2 (se) alleles in which 9.3 kb containing the entire coding region of FUT2 is deleted by Alu-mediated nonhomologous recombination. In addition to this allele, three SNPs of FUT2, c.375A>G, c.385A>T, and c.571C>T, appear to be prevalent in certain Oceanian populations such as Polynesians. Recently, we developed an endpoint genotyping assay to determine the sedel2 zygosity, using a FAM-labeled probe for detection of the sedel2 allele and a VIC-labeled probe for detection of FUT2. In this study, instead of the VIC probe, a HEX-labeled probe covering both c.375A>G and c.385A>T and a Cy5-labeled probe covering c.571C>T were added to the sedel2 allele assay mixture to allow simultaneous detection of these four variations by endpoint genotyping for the sedel2 zygosity and fluorescence melting curve analysis for c.375A>G, c.385A>T, and c.571C>T genotyping. The results obtained from 24 Samoan subjects with this method were identical to those obtained with previous methods. Therefore, it appears that present method can accurately determine these four variations simultaneously.

Current paper reviews applications of luminescence bioassays for monitoring low-intensity factors, namely, radioactivity of different types (alpha, beta and gamma), and bioactive compounds (humic substances and fullerenols). Luminescence intensity is taken as a physiological parameter of luminous organisms. High rates of luminescence response can provide (1) a proper number of experiments under comparable conditions and, therefore, proper statistical processing, with this being highly important for ‘noisy’ low-dose exposures; (2) non-genetic, i.e. biochemical and physicochemical mechanisms of cellular response, in accordance to “exposome” concept. Bioassays based on luminous marine bacteria, their enzymes, and fluorescence coelenteramide-containing proteins were used to compare results of low-intensity exposures at cellular, biochemical and physicochemical levels, respectively. Results of the cellular exposures were discussed in terms of hormesis concept. Bioluminescence time dependence under low-dose radiation exposures corresponded to hormesis or threshold models; no bioluminescence monotonic dependency on intensity of exposure (dose rate, radioactivity, concentration) was found. Bioluminescence activation and absence of its dependency on intensity of exposure can be accepted as features of cellular adaptive response. Changes of biological luminescence were analyzed and discussed for bioassays of lower organization level – enzymes and florescent protein.

Although cofactor regeneration is an established system in biocatalysis, work remains in developing new and alternative cofactor regeneration systems with greater efficiency, ease of use, and higher atom economy. In addition, cofactor regeneration system only works if the cofactor regeneration reaction operates at similar kinetics compared to the biotransformation reaction. This meant that only specific cofactor regeneration system is capable of coupling with particular biotransformation reaction. This then leaves open the field for the development of a plethora of alternative cofactor regeneration systems each capable of coupling with different biotransformation reaction of different kinetics. This short write-up examines the possibility of tapping on the NADH regenerated from a two-step ethylene glycol utilization pathway. Current knowledge suggests that this angle has not been explored; thereby, opening up possibilities for future experimental investigations into the feasibility of coupling ethylene glycol utilization pathway with biotransformation reaction as a coupled cofactor regeneration system.

Medical imaging is an active field of research that fosters the necessity for novel multimodal imaging probes. In this line, nanoparticle-based contrast agents are of special interest, since those can host functional entities either within their interior, reducing potential toxic effects of the imaging tracers, and on their surface, providing high payloads of probes, due to their large surface-to-volume ratio. The long-term stability of the particles in solution is an aspect usually under-tackled during probe design in research laboratories, since their performance is generally tested briefly after synthesis. This may jeopardize a later translation into practical medical devices, due to stability reasons. To dig into the effects of nanoparticle aging in solution, respect to their behavior in vivo, iron oxide stealth nanoparticles were used at two stages (3 weeks vs. 9 months in solution), analyzing their biodistribution in mice. Both sets of nanoprobes showed similar sizes, zeta potentials and morphology, as observed by DLS and TEM but, fresh nanoparticles accumulated in the kidneys after systemic administration, while aged ones accumulated in liver and spleen, confirming an enormous effect of particle aging on their in vivo behavior, despite barely noticeable changes perceived on a simple inspection of their structural integrity.

Development of novel anti-cancer peptides requires a rapid screening process which can be accelerated by using appropriate in vitro tumor models. Breast carcinoma tissue is a three dimensional (3D) microenvironment, which contains a hypoxic center surrounded by dense proliferative tissue. Biochemical clues provided by such 3D cell mass cannot be recapitulated in conventional 2D culture systems. In this experiment, we evaluate the efficacy of the sandalwood peptide, cyclosaplin on established in vitro 3D silk breast cancer model using invasive MDA-MB-231 cell line. The anti-proliferative effect of the peptide on 3D silk tumor model is monitored by alamar blue assay, with conventional 2D culture as control. The proliferation rate, glucose consumed, LDH, and MMP-9 activity of Human breast cancer cells are higher in 3D constructs compared to 2D. A higher concentration of drug is required to achieve 50% cell death in 3D culture than 2D cultures. The cyclosaplin treated MDA-MB-231 cells showed significant decrease in MMP-9 activity in 3D constructs. Microscopic analysis revealed the formation of cell clusters evenly distributed in the scaffolds. The drug treated cells were less in number, smaller and showed unusual morphology. Overall, these findings indicate the role of cyclosaplin as a promising anti-cancer therapeutics.

Pathogenic microorganisms can cause a range of infectious diseases and are often treated with antimicrobial agents. The rise of antimicrobial resistance caused by inappropriate use of these agents in various settings has become a global health threat. There is, therefore, a need to find new and effective agents to combat infections caused by resistant pathogens. Nanotechnology has seen significant growth and development in recent years and has a wide range of applications in various fields, including health, agriculture, and industry. This novel area offers the potential for the synthesis of nanoparticles (NPs) with antimicrobial activity, such as iron oxide nanoparticles (IONPs). The use of IONPs is a promising way to overcome antimicrobial resistance or pathogenicity because of their ability to interact with several biological molecules and to inhibit microbial growth. This review focuses on the most common green synthesis methods for producing IONPs using bacteria, fungi, plants, and organic waste, as well as the most common methods for the characterization of these IONPs. Finally, it summarizes their application as promising antibacterial, antifungal, antiparasitic, and antiviral agents.

Cistus incanus L. (CI) has been proposed as an innovative functional supplement of food products, and hence the present study aimed to evaluate the effect of the addition of dried CI on the properties of bread. Bread was prepared from white wheat flour supplemented with the addition of 1%, 2%, 3%, 4%, and 5% of ground CI. After the completion of baking process, various characteristics of the obtained bread product, such as yield, volume, porosity, acidity, color, and texture, were evaluated. In addition, total phenolic content (TPC), ABTS radical scavenging activity, CHEL chelating power, and ability to quench OH∙ radicals were measured. The results showed that the addition of CI to bread caused a reduction in the volume of bread, but texture of the crumbs was acceptable. Acidity and moisture content of bread were found to be increased following CI enrichment. Significant changes in the ash content and the color of bread crumbs were also observed. Bread incorporated with CI was characterized by significantly higher TPC and much higher antioxidant activity, as measured by ABTS, CHEL, and OH∙ radicals, compared to control bread. Supplementation of bread with 3% CI produced a product with desirable characteristics which was also favored by consumers.

Bioethanol has many environmental and practical benefits as a transportation fuel. It is one of the best alternatives to replace fossil fuels due to its liquid nature which is similar to petrol and diesel fuels traditionally used in transportation. In addition, bioethanol production technology has the capacity for negative carbon emissions which is vital for solving the current global warming dilemma. However, conventional bioethanol production takes place based on an inland site and relies on freshwater and edible crops (or land suitable for edible crop production) for production, which has led to the food vs fuel debate. Establishing a coastal marine biorefinery (CMB) system for bioethanol production that is based on coastal sites and relies on marine resources (seawater, marine biomass and marine yeast) could be the ultimate solution. In this paper, we aim to evaluate the environmental impact of using seawater for bioethanol production at coastal locations as a step towards the evaluation of a CMB system. Hence, a life cycle assessment for bioethanol production was conducted using the proposed scenario named Coastal-Seawater and compared to the conventional scenario, named Inland-Freshwater (IF). The impact of each scenario in relation to climate change, water depletion, land use and fossil depletion was studied for comparison. The coastal-seawater scenario demonstrated an improvement upon the conventional scenario in all the selected impact categories. In particular, the use of seawater in the process had a significant effect on water depletion showing an impact reduction of 31.2%. Furthermore, reductions are demonstrated in natural land transformation, climate change and fossil depletion of 5.5%, 3.5% and 4.2% respectively. This indicates the positive impact of using seawater and coastal locations for bioethanol production and encourages research to investigate the CMB system.

An upsurge in global population over the years and rapid urbanization have accelerated huge dependence on petroleum-derived fuels and consequent environment concerns owing to green-house gas emissions in the atmosphere. An integrated biorefinery uses lignocellulosic feedstock as raw material for the production of renewable biofuels, and other fine chemicals. The sustain-able bio-economy and the biorefinery industry would benefit greatly from the effective use of lignocellulosic biomass obtained from agricultural feedstocks to replace petrochemical products. Lignin, cellulose, hemicellulose, and other extractives, which are essential components of ligno-cellulosic biomass, must be separated or upgraded into useful forms in order to fully realize the potential of biorefinery. The development of low-cost and green pretreatment technologies with effective biomass deconstruction potential is imperative for an efficient bioprocess. The abun-dance of microorganisms along with their continuous production of various degradative en-zymes makes them suited for the environmentally friendly bioconversion of agro-industrial wastes into viable bioproducts. The present review highlights the concept of biorefinery, ligno-cellulosic biomass and its valorization by green pretreatment strategies into biofuels and other biochemicals. The major barriers and challenges in bioconversion technologies, environmental sustainability of the bioproducts and promising solutions to alleviate those bottlenecks are also summarized.

Yeasts belonging to the Metschnikowia genus are particularly interesting for the unusual formation of only two needle-shaped ascospores during their mating cycle. Presently, the meiotic process that can lead to only two spores from a diploid zygote is poorly understood. The expression of fluorescent nuclear proteins should allow the meiotic process to be visualized in vivo; however, no large-spored species of Metschnikowia has ever been transformed. Accordingly, we aimed to develop a transformation method for Metschnikowia borealis, a particularly large-spored species of Metschnikowia, with the goal of enabling the genetic manipulations required to study biological processes in detail. Genetic analyses confirmed that M. borealis, and many other Metchnikowiacea, are CUG-Ser yeasts. Codon-optimized selectable markers lacking CUG codons were used to successfully transform M. borealis by electroporation and lithium acetate, and transformants appeared to be the result of random integration. Mating experiments confirmed that transformed-strains were capable of generating large asci and undergoing recombination. Finally, random integration was used to transform an additional 18 yeast strains, and all attempts successfully generated transformants. The results provide a simple method to transform many yeasts from an array of different clades and can be used to study or develop many species for various applications.

Microglial cells, the resident macrophages of the CNS, exist in a process-bearing, ramified/surveying phenotype under resting conditions. Upon activation by cell damaging factors they get transformed to an amoeboid phenotype releasing various cell products including pro-inflammatory cytokines, chemokines, proteases, reactive oxygen/nitrogen species and the excytotoxic ATP and glutamate. In addition, they engulf pathogenic bacteria or cell debris and phagocytose them. However, already resting/surveying microglia has a number of important physiological functions in the CNS; they e.g. shield small disruptions of the blood-brain barrier by their processes, dynamically interact with synaptic structures and clear surplus synapses during development. In neurodegenerative illnesses they aggravate the original disease by a microglia-based compulsory neuroinflammatory reaction. Therefore, the blockade of this reaction improves the outcome of Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, amyotrophic lateral sclerosis, etc. The function of microglia is regulated by a whole array of purinergic receptors classified as P2Y12, P2Y6, P2Y4, P2X4, P2X7, A2A, A3, and being targets for endogenous ATP, ADP, or adenosine. ATP is sequentially degraded by the ecto-nucleotidases and 5’-nucleotidase enzymes to the weak adenosine agonist inosine as an end-product. The appropriate selective agonists/antagonists for purinergic receptors as well as the respective enzyme inhibitors may profoundly interfere with microglial functions and reconstitute the homeostasis of the CNS disturbed by neuroinflammation.

In the sugar industry, dextran generates difficulties in the manufacturing process. Crude dextranase (EC 3.2.1.11) to eliminate dextran in sugar is an effective practice. In this study, a synthetic dextranase encoding gene of the filamentous fungus Talaromyces minioluteus, lacking its putative native signal peptide (1-20 amino acids) and the next 30 amino acids (r-TmDEX49A-ΔSP-ΔN30), was fused to the Saccharomyces cerevisiae prepro -factor (MF-2) signal sequence and expressed in Komagataella phaffii under the constitutive GAP promoter. K. phaffii DEX49A-ΔSP-ΔN30, constitutively producing and secreting the truncated dextranase was obtained. The specific activity of the truncated variant resulted nearly the same in relation to the full-length mature enzyme (900-1000 U.mg-1 of protein). At shaker scale (100 mL) in YPG medium, the enzymatic activity was 273 U.mL-1. The highest production level was achieved in a fed-batch culture (30 h) at 5 L fermenter scale using the FM21-PTM1 culture medium. The enzymatic activity in the culture supernatant reached 1614 U.mL-1 and the productivity was 53800 U.L-1.h-1 (53.8 mg.L-1.h-1), the highest reported so far for a DEX49A variant. Dextran decreased r-TmDEX49A-ΔSP-ΔN30 mobility in affinity gel electrophoresis, providing evidence of carbohydrate-protein interactions. K. phaffii DEX49A-ΔSP-ΔN30 shows great potential as a methanol-free, commercial dextranase production system.

The following study reports on the first thraustochytrid isolates identified from Iceland. They were collected from three different locations off the northern coast of the country (Location A, Skagaströnd; Location B, Hveravík; and Location C, Eyjafjörður). Using 18S rDNA sequence analysis, isolates from Locations A and B were identified within the Thraustochytrium kinnei species while other isolates within the Sicyoidochytrium minutum species when compared to other known strains. Cells isolated from Locations A (2.10 ± 0.70 g/L) and B (1.54 ± 0.17 g/L) produced more biomass than the ones isolated from Location C (0.43 ± 0.02 g/L). This study offers the first-time examination of the utility of byproducts from fisheries as a nitrogen source in media formulation for thraustochytrids. Experiments showed that isolates produced more biomass (per unit of substrate) when cultured on nitrogen of marine (2.55 ± 0.74 g/L) as compared to of commercial origin (1.06 ± 0.57 g/L). Glycerol (2.43 ± 0.56 g/L) was a better carbon source than glucose (1.84 ± 0.57 g/L) in growth studies. Fatty acid (FA) profiles showed that the isolates from Location C (S. minutum) had low ratios of monounsaturated (4.21 ± 2.96%) and omega-6 (0.68 ± 0.59%) FAs. However, the isolates also had high ratios of docosahexaenoic acid (DHA; 35.65 ± 1.73%) and total omega-3 FAs (40.39 ± 2.39%), indicating that they could serve as a source of marine oils for human consumption and in aquaculture feeds. The T. kinnei isolates from Location A could be used in biodiesel production due to their high ratios of monounsaturated (18.38 ± 6.27%) long chain (57.43 ± 8.27%) FAs.

Plant diseases are among the major factors affecting plant productivity. Biological control of plant diseases is preferred over chemical control as it is environment-friendly, cost-effective, and sustainable. Among many microbes capable of providing biological control of plant diseases, probiotic Bacillus species are most promising as they can survive in adverse conditions, provide plants with a wide range of benefits including protection from phytopathogens. Wheat blast caused by Magnaporthe oryzae Triticum pathotype (MoT) has emerged as a potential threat to global wheat production. Due to unreliability of fungicides and limited cultivar resistance, we aimed to screen and identify potential antagonist bacteria collected from internal tissues of rice and wheat seeds to determine their in vitro and in vivo inhibitory effects against MoT. Dual culture and seedling assays were performed to evaluate the efficacy of probiotic bacteria. Out of 170 bacterial isolates, three bacteria (BTS-3, BTS-4, and BTLK6A) were screened as potential antagonists against MoT in vitro. Artificial inoculation at the seedling stage showed that the isolates BTS-4, BTS–3, and BTLK6A reduced 89, 88, and 85% of wheat blast disease severity, respectively, compared to mock-inoculated control. The bacterial isolates were identified as Bacillus subtilis (BTS-3) and B. velezensis (BTS-4 and BTLK6A) through genome phylogeny. The whole genome sequence of these three bacterial strains decoded a number of orthologs to intrinsic genes of antimicrobial peptides, antioxidant defense enzymes, cell wall degrading enzymes, compounds involved in the induction of systemic resistance (ISR) in host plants, and volatile compounds to make them promising biologicals to control MoT in wheat. Combined data of in vitro and in vivo along with genome analysis suggest that Bacillus spp. suppress the destructive wheat blast disease likely through antibiosis and ISR in the host plants. Further field evaluation and characterization of antimicrobial compounds are needed for a better understanding of the mode of action and practical recommendation of these bacteria for wheat blast control in the farmers’ fields.

In the last decade, the genetic engineering world has been shaken up by a relatively new genetic editing tool based on RNA-guided Nucleases (RGNs): the CRISPR/Cas9 system. Since the first report in 1987 and its characterization in 2007 as a bacterial defense mechanism, the interest and research on this system have grown exponentially. CRISPR systems provide immunity to bacteria against invading genetic material; however, with specific modifications in sequence and structure, it becomes a precise editing system that makes it possible to genetically modify almost any organism. There are diverse approaches regarding the refinement of these modifications, such as constructing more accurate nucleases, understanding the cellular context and facing the epigenetic conditions, or re-designing guide RNAs (gRNAs). Considering the critical importance for the correct CRISPR/Cas9 systems performance, our scope will emphasize in the latter approach. Hence, we present an overview of the past and the most recent guide RNA web-based design tools, highlighting their computational architecture and gRNA characteristics evolution through the years. Our study concisely explains the computational approaches that use machine learning techniques, deep neural networks, and large datasets of gRNA/target interactions to make possible both predictions and classifications directed to design, optimize, and create promising gRNAs suitable for future gene therapies.

We know that living matter must behave in accordance with the universal laws of physics and chemistry. However, these laws are insufficient to explain the specific characteristics of the vital phenomenon and, therefore, we need new principles, intrinsic to biology, which are the basis for developing a theoretical framework for understanding life. Here I propose what I call the seven commandments of life (the Vital Order, the Principle of Inexorability, the Central Dogma, the Tyranny of Time, the Evolutionary Imperative, the Conservative Rule, the Cooperating Thrust) as a set of principles that help us explain the vital phenomenon from an evolutionary perspective. In a metaphorical way, we can consider life like an endless race in which living beings are the runners, who are changing as the race goes on (the evolutionary process), and the commandments the rules.

The RNA-guided endonuclease system CRISPR-Cas9 has been extensively modified since its discovery, allowing its capabilities to be extending far beyond double-stranded cleavage to high fidelity insertions, deletions, and single base edits. Such innovations have been possible due to the modular architecture of CRISPR-Cas9 and the robustness of its component parts to modifications and the fusion of new functional elements. Here, we review the broad toolkit of CRISPR-Cas9-based systems now available for diverse genome editing tasks. We provide an overview of their core molecular structure and mechanism and distil the design principles used to engineer their diverse functionalities. We end by looking beyond the biochemistry and towards the societal and ethical challenges that these CRISPR-Cas9 systems face if their transformative capabilities are to be deployed in a safe and acceptable manner.

Contamination of the environment by petroleum products is a growing concern worldwide, and strategies to remove these contaminants have been evaluated. One of these strategies is biodegradation, which consists of the use of microorganisms. Biodegradation is significantly improved by increasing the temperature of the milieu, thus, the use of thermophiles, microbes that thrive in high-temperature environments, will render this process more efficient. For instance, various thermophilic enzymes have been used in industrial biotechnology because of their unique catalytic properties. Biodegradation has been extensively studied in the context of mesophilic microbes, and the mechanisms of biodegradation of aliphatic and aromatic petroleum hydrocarbons have been elucidated. However, in comparison, little work has been carried out on the biodegradation of petroleum hydrocarbons by thermophiles. In this paper, a detailed review of the degradation of petroleum hydrocarbons (both aliphatic and aromatic) by thermophiles has been carried out. This work has identified the characteristics of thermophiles, and unravelled specific catabolic pathways of petroleum products that are only found in thermophiles. Gaps that limit our understanding of the activity of these microbes have also been highlighted, and finally, different strategies that can be used to improve the efficiency of degradation of petroleum hydrocarbons by thermophiles have been proposed.

Sleep quality has been directly linked to cognitive function, quality of life, and a variety of serious diseases across many clinical domains such as psychiatry and cardiology. Standard methods for assessing sleep involve overnight studies in hospital settings, which are uncomfortable, expensive, not representative of real sleep, and difficult to conduct on a large scale. Recently, a number of commercial digital devices have been developed that record physiological data which can act as a proxy for sleep quality in lieu of standard electroencephalogram recording equipment. Each device company makes different claims of accuracy and measures different features of sleep quality, and it is still unknown how well these devices correlate with one another and perform in a research setting. In this pilot study of 21 participants, we investigated whether outputs from four sensors, specifically FitBit, Withings Aura, Hexoskin, and Oura Ring, were related to known cognitive and psychological metrics, including the PSQI and N-back test. We found that sleep metrics extracted from these devices did not predict cognitive and psychological metrics well in our pilot data. However, we did identify certain signification associations, specifically the Oura Ring’s total sleep duration and efficiency in relation to the PSQI measure with p=0.004 and p=0.033, respectively. Additionally, correlation of various sleep features among the devices across the sleep cycle was almost uniformly low. These findings can hopefully be used to guide future sensor-based sleep research.

Food-grade titanium dioxide (E171) contains variable percentages of TiO2 nanoparticles (NPs) posing awareness about its potential effects on human and animal health. Despite many studies, the actual relationship between the physicochemical properties of E171 NPs and their interaction with biological targets is still far from clear. We evaluated the acute toxicity of E171 in invertebrate and vertebrate animals. In the nematode C. elegans, E171 up to 1.0 mg/mL did not affect worm’s viability and lifespan but significantly impaired pharyngeal function, reproduction, and development. We also focused our attention on the fate of E171 after its penetration inside the circulatory tree, investigating whether its intravenous administration to mice could result in an acute over-absorption to filter organs. A significant increase of hepatic Ti concentration and the formation of microgranulomas were observed. Interstitial inflammation and parenchymal modification were found in lung coupled with Ti accumulation, probably due to the propensity of TiO2 NPs to agglomerate, as demonstrated by transmission electron microscopy experiments showing that the incubation of E171 with serum promoted the formation of compact clusters. Overall, these data emphasize, once again, the actual risk for human and animal exposure to E171.

We report a modification of the freeform reversible embedding of suspended hydrogels (FRESH) 3D printing method for the fabrication of freeform perfusable microfluidics inside a hydrogel matrix. Xanthan gum is deposited into a CaCl2 infused gelatin slurry to form filaments, which are consequently rinsed to produce hollow channels. This provides a simple method for rapid prototyping of microfluidic devices based on biopolymers and potentially a new approach to the construction of vascular grafts for tissue engineering.

PD-1/PD-L1 axis is one key therapeutic target against tumor cell immune escape. Structurally essential to the PD-1/PD-L1-linked immune escape is the binding interface of the PD-1/PD-L1 complex structure. Incorporating currently available PD-1/PD-L1-related experimental structures, this article unveils two sets of experimentally observed inter-molecular electrostatic interactions which stabilize the binding interface of the PD-1/PD-L1 complex structure. For the first time, this article proposes an evolutionary structural hypothesis that, as a result of natural selection, PD-1 is able to genetically mutate itself to structurally disrupt the PD-1/PD-L1 axis towards the restoration of T cell-mediated anti-tumor immunity.

This work describes practical immunization of European sea bass (Dicentrarchus labrax) juveniles against viral nervous necrosis virus (VNNV), a betanodavirus causing worldwide mortalities in many fish species. Protection was obtained with the so called spinycterin vehicles consisting in irreversibly DNA-damaged DNA-repair-less E.coli displaying at their surface a downsized antigen. In this work we, i) maximized bacterial expression levels by downsizing the C protein to a fragment (frgC91-220) containing most of its antigenicity, ii) developed an scalable autoinduction bacterial media based in soy-bean increasing membrane display and reproducibility, iii) enriched surface expression by screening different anchors from several prokaryotic origins (anchor+frgC91-220), iv) preserved frgC91-220 antigenicity by inactivating bacteria by irreversible DNA-damage by means of Ciprofloxacin, and v) increased safety using a repair-less E.coli strain as spinycterin chassis. These second generation of spinycterins protected fish against VNNV challenge with partial (Nmistic+frgC91-220) or 100 % (YBEL+frgC91-220 ) protection, in contrast to those fish immunized with frgC91-220 spinycterins. The proposed spinycterin platform has high levels of environmental safety and cost effectiveness, thus providing potential for small fish vaccines for sustainable aquaculture.

Garlic and onion are either cooked like a vegetable because of their green leaves or are also used as a condiment. Many scientific studies affirm the positives of both for their anti-inflammatory, antioxidant, and antimicrobial potential. Moreover, garlic and onion are regularly employed to treat cardiovascular illnesses, strokes, atherosclerosis, hypertension, hyperlipidemias, and thrombosis, and are also proved effective against Alzheimer's, diabetes, and cancers. Here we have compiled a piece of information regarding the compounds present in garlic and onion along with their pharmacological properties. Although much more studies are required to refine the utilization and enhance garlic and medicine's effectiveness. We hope this work will provide helpful information regarding their pharmacological aspects.

We present a set of software packages that provide uniform access to diverse biological vocabulary resources that are instrumental for current biocuration efforts and tools. The Unified Biological Dictionaries (UniBioDicts or UBDs) provide a single query-interface for accessing the online API services of leading biological data providers. Given a search string, UBDs return a list of matching term, identifier and metadata units from databases (e.g. UniProt), controlled vocabularies (e.g. PSI-MI), and ontologies (e.g. GO, via BioPortal). This can be coupled to for instance the ‘vsm-autocomplete’ module: an input field (user-interface component) that offers autocomplete lookup for these dictionaries. UBDs create a unified gateway for accessing life science concepts, helping curators find annotation terms across resources (based on descriptive metadata and unambiguous identifiers), and data users search and retrieve the right query terms.

Multicellular life forms have evolved many times in our planet, suggesting that this is a common evolutionary innovation. Multiple advantages have been proposed for multicellularity (MC) to emerge. In this paper we address the problem of how the first precondition for multicellularity, namely "stay together" might have occurred under spatially limited resources exploited by a population of unicellular agents. Using a minimal model of evolved cell-cell adhesion among growing and dividing cells that exploit a localised resource with a given size, we show that a transition occurs at a critical resource size separating a phase of evolved multicellular aggregates from a phase where unicellularity (UC) is favoured. The two phases are separated by an intermediate domain where where both UC and MC can be selected by evolution. This model provides a minimal approach to the early stages that were required to transition from Darwinian individuality to cohesive groups of cells associated with a physical cooperative effect: when resources are present only in a localised portion of the habitat, MC is a desirable property as it helps cells to keep close to the available local nutrients.

Background: Succinate dehydrogenase (SDH)-deficient renal cell carcinoma (RCC) is mainly associated with SDHB gene mutation and sometimes with SDHC or SDHD mutation. However, only three case of SDHA-deficient RCC have been reported, and the relation of SDHA mutation to RCC has not been clarified. Methods: We investigated SDHA/B/C/D gene mutations in 72 human RCCs by targeted next-generation sequencing, and also assessed SDHA and nuclear factor erythroid 2–related factor 2 (Nrf2) protein expression in 30 tumors. Results: NGS revealed SDHA gene mutations associated with amino acid sequence variations in 12 tumors, while no tumors had SDHB/C/D mutations. The SDHA gene mutations were identical in somatic and germline DNA of all 12 patients. Tumors with SDHA mutations showed a decrease of SDHA protein (p = 0.0177) and an increase of Nrf2 protein (p = 0.0120), with an inverse correlation between SDHA and Nrf2 protein expression (p = 0.0321). Tumor cells with SDHA mutations characteristically had eosinophilic cytoplasm and showed various patterns of proliferation. Conclusions: Our observations suggested that germline SDHA gene mutations might be linked to hereditary RCC associated with mitochondrial dysfunction.

Immobilized metal affinity chromatography (IMAC) is a popular and valuable method for the affinity purification of polyhistidine-tagged recombinant proteins. However, it often shows practical limitations, which might require cumbersome optimizations, additional polishing, and enrichment steps. Here, we present functionalized corundum particles for the efficient, economical, and fast purification of recombinant proteins in a column-free format. The corundum surface is first derivatized with the amino silane APTES, then EDTA dianhydride, and subsequently loaded with nickel ions. The Kaiser test, well-known in solid-phase peptide synthesis, was used to monitor amino silanization and the reaction with EDTA dianhydride. In addition, ICP-MS was performed to quantify the metal-binding capacity. His-tagged protein A/G (PAG), mixed with bovine serum albumin (BSA), was used as a test system. The PAG binding capacity was around 3 mg protein per gram of corundum. Cytoplasm obtained from different E. coli strains was examined as an example of a complex matrix. The imidazole concentration was varied in the loading and washing buffers. As expected, higher imidazole concentrations during loading are usually beneficial when higher purities are desired. Even when higher sample volumes, such as one liter, were used, recombinant protein down to a concentration of 1 µg/mL could be isolated selectively. Comparing the corundum material with standard Ni-NTA agarose beads indicated higher purities of proteins isolated using corundum. His6-MBP-mSA2, a fusion protein consisting of monomeric streptavidin and maltose-binding protein in the cytoplasm of E. coli, was purified successfully. To show that this method is also suitable for mammalian cell culture supernatants, purification of the SARS-CoV-2-S-RBD His8 expressed in human Expi293F cells was performed. The material cost of the nickel-loaded corundum material (without regeneration) is estimated to be less than 30 cents for 1 gram of functionalized support or 10 cents per milligram of isolated protein. Another advantage of the novel system is the corundum particles' extremely high physical and chemical stability. The new material should be applicable in small laboratories and large-scale industrial applications. In summary, we could show that this new material is an efficient, robust, and cost-effective purification platform for the purification of His-tagged proteins, even in challenging, complex matrices and large sample volumes of low product concentration.

Understanding the molecular underpinnings of the evolution of complex (multi-part) systems is a fundamental topic in biology. One unanswered question is the extent to which similar or different genes and regulatory interactions underlie similar complex systems across species. Animal eyes and phototransduction (light detection) are outstanding systems to investigate this question because some of the genetics underlying these traits are well-characterized in model organisms. However, comparative studies using non-model organisms are also necessary to understand the diversity and evolution of these traits. Here, we compare the characteristics of photoreceptor cells, opsins, and phototransduction cascades in diverse taxa, with particular focus on cnidarians. In contrast to the common theme of deep homology, whereby similar traits develop mainly using homologous genes, comparisons of visual systems - especially in non-model organisms - are beginning to highlight a “deep diversity” of underlying components, illustrating how variation can underlie similar complex systems across taxa. Although using candidate genes from model organisms across diversity was a good starting point to understand the evolution of complex systems, unbiased genome-wide comparisons and subsequent functional validation will be necessary to uncover unique genes that comprise complex systems of non-model groups to better understand biodiversity and its evolution.

The genus Prototheca, unicellular, non-photosynthetic, yeast-like microalgae, is a pathogen of concern for the dairy industry causing bovine mastitis that currently cannot be cured and hence generates significant economic losses in milk production. In this study, for the first time in Ecuador, we identify Prototheca bovis as the etiologic agent of chronic mastitis in dairy cattle. Milk samples (n=458) of cows with chronic mastitis were cultured on Sabouraud Dextrose Agar (SDA). Microscopy and cytB gene sequencing were used to identify Prototheca, whereby Prototheca bovis was isolated from 15.1% (n=69) of the milk samples, one of the highest infection rates that can be found in the literature in a “non-outbreak” situation. No other Prototheca species were found. We were unable to isolate the alga from environmental samples. We showed that P. bovis was relatively resistant to disinfectants used to sterilize milking equipment on the cattle farms where it was isolated. We discuss how to avoid future infection and also hypothesize that the real prevalence of Prototheca infection in bovine mastitis is probably much higher than what was detected. We recommend a protocol to increase the diagnostic yield in the bacteriology laboratory.

Superantigens are unconventional antigens which recognise immune receptors outside the usual binding sites e.g. complementary determining regions (CDRs), to elicit a response within the target cell. T-cell superantigens crosslink T-cell receptors and MHC Class II molecules on antigen-presenting cells, leading to lymphocyte recruitment, induction of cytokine storms and T-cell anergy or apoptosis among many other effects. B-cell superantigens, on the other hand, bind immunoglobulin receptors on B-cells affecting opsonisation, IgG-mediated phagocytosis, and drive B-cells into apoptosis. Here, through a review of the structural basis for recognition of immune receptors by superantigens, we show that their binding interfaces share specific physicochemical characteristics when compared with other protein-protein interaction complexes. Given that antibody-binding superantigens have been exploited extensively in industrial antibody purification, these observations could facilitate further protein engineering to optimize the use of superantigens in this and other areas of biotechnology.

The origin of the nucleus remains a great mystery in life science, although nearly two centuries have passed since the discovery of nuclei. To date, studies of eukaryogenesis have focused largely on micro-evolutionary explanations. Here, we examined macro-patterns of C-values (the total amount of DNA within the haploid chromosome set of an organism) for over 110,000 species and the chromosome numbers for over 11,000 species and their potential links with the state of atmospheric oxidation over geological time. Eukaryogenesis was in sync with an over 2.5 order-of-magnitude increase in genome size from prokaryote to eukaryote, and also with a rapid rise of atmospheric oxidation, suggesting that eukaryogenesis would have resulted from a regime shift of genomes driven by the oxidation-driven complexification and structuralization (e.g. chromatin packing).

Polysaccharides from marine algae exhibit beneficial biological activities. In this study, we examined the effect of polysaccharides from Codium fragile (PCF) on prebiotic and anti-obesity activity. PCF increases the growth of specific beneficial microbial populations with concomitant decrease in pathogenic microbes. Further, total phenolic content (TPC), total flavonoid content (TFC), and DPPH radical scavenging activity (DPPH activity) after fermentation with PCF as carbon source were higher than for the glucose as control. Moreover, PCF inhibited adipocyte differentiation by inducing differentiation-related factors when the induction of 3T3-L1 preadipocytes into adipocytes was induced. Therefore, we suggest that PCF can be used as prebiotic material with anti-obesity for human health.

Notwithstanding the advances achieved in the last decades in the field of synthetic bone substitutes, the development of biodegradable 3D scaffolds with ideal mechanical and biological properties remains an unattained challenge. In this work, a novel approach is explored to produce synthetic bone grafts mimicking the complex bone structure using additive manufacturing. For the first time, scaffolds were produced, using an extrusion technique, composed of a thermoplastic polymer, polycaprolactone (PCL), hydroxyapatite nanoparticles (HANp), and polyethylene glycol diacrylate (PEGDA). These scaffolds were further compared with two groups of scaffolds: one composed of PCL and another of PCL and HANp. After production, optimisation and characterisation of these scaffolds, an in vitro evaluation was performed using human dental pulp stem/stromal cells (hDPSCs). Through the findings it was possible to conclude that PEGDA scaffolds were successfully produced presenting networks of interconnected channels, presenting hydrophilic properties (15.15 4.06°), adequate mechanical performance (10.41MPa 0.934), and allowing a cell viability significantly superior to the other groups analysed. To conclude, findings in this study demonstrated that PCL, HANp and PEGDA scaffolds may have promising effects on bone regeneration and might open new insights for 3D tissue substitutes.

White adipose tissue (WAT) is distributed in several depots that have distinct metabolic and inflammatory functions. In our body there are subcutaneous (sWAT), visceral (vWAT) and bone marrow fat depots (BFAT). Obesity affects size, function and inflammatory state of WATs. This process can alter the stem cell niches present in these tissues and affect the functions of stem cells residing within. In particular, obesity may affect the activity of mesenchymal stromal cells (MSCs) present in WAT. MSCs are an heterogenous population containing stromal cells, progenitor cells, fibroblasts and stem cells that are able to differentiate in adipocytes, chondrocytes, osteocytes and other mesodermal derivatives. We performed a comparison of the effects of obesity on MSCs obtained from sWAT, vWAT and BFAT. Our study evidenced that obesity affected mainly the biological functions of MSCs obtained from bone marrow and vWAT with a decrease in proliferation rate, reduced percentage of cells in S phase and trigger of senescence. The onset of senescence was confirmed by expression of genes belonging to RB and P53 pathways. Our study evidenced that negative consequences of obesity on body physiology may be related also to impairment in the functions of stromal compartment present in the several adipose tissues. This finding provides new insights on the targets that should be considered for an effective treatment of obesity-related diseases.

Fruit production is an important part of the gross domestic product for many countries around the world especially to those who have a strong focus on agriculture. However, long-term maintenance and yield stability of fruit production may be threatened by the ongoing climate change and its consequences like extended drought periods, heavy rain events, and floodings. Genome editing, with its progressive technological developments, offers opportunities to adapt relevant fruit plant species to new climatic conditions. Among modern genome editing techniques, CRISPR/Cas, in particular, has the potential to support breeding for those fruit plant species with extended breeding cycles, e.g., perennial fruits. In this review, we discuss CRISPR/Cas and other genome editing techniques in detail and how these techniques can be applied to support the breeding of fruit plant species for adaptation to changing climates. The chronological history of CRISPR/Cas9 systems, their associated computational tools, genomic data sources, transformation methods along with their delivery vehicles, quality improvement, environmental-stress resiliency, limitations, and future perspectives will also be discussed with respect to securing future global fruit production.

: Coffee is one of the most consumed beverages in the world; its production is based mainly on varieties of the Coffea arabica species. Mexico stands out for its specialty and organic coffee. In Guerrero the production is given by small indigenous community’s cooperatives that market their product as raw material. Official Mexican Standards stipulate the necessary requirements for its commercialization within the national ter-ritory. In this work, the physical, chemical, and biological characterizations of green, medium, and dark roasted beans from C. arabica varieties were carried out. Analysis of HPLC showed higher chlorogenic acid (55 mg/g) and caffeine (1.8 mg/g) contents in the green beans of the Bourbon and Oro Azteca varieties. The caffeine (3.88 mg/g) and melanoidins (97 and 29 mg/g) contents increased according to the level of roasting; a dissimilar effect was found in the chlorogenic acid content (14.5 mg/g). The adequate nutritional content and the sensory evaluation allowed the classification of dark roasted as premium coffee (84.25 points) and the roasted medium as specialty coffee (86.25 points). The roasted coffees presented antioxidant activity without cytotoxic effect. The results obtained will serve as a basis for making decisions on the improvements of the coffees analyzed.

Here we propose the concept of an electro–microbial route to uncouple food production from photosynthesis, thereby enabling production of nutritious food in space without the need to grow plant-based crops. In the proposed process, carbon dioxide is fixed into ethanol using either chemical catalysis or microbial carbon fixation, and the ethanol created is used as a carbon source for yeast to synthesize food for human or animal consumption. The process depends upon technologies that can utilize electrical energy to fix carbon into ethanol and uses an optimized strain of the yeast Saccharomyces cerevisiae to produce high quality, food grade single cell protein using only ethanol, urea, phosphate, and inorganic salts as inputs. Unlike crops using photosynthesis that require months to mature and are challenging to grow under the conditions found in space, the electro–microbial process could generate significant quantities of food on demand with potentially high yields and productivities. In this paper we explore the potential of the proposed process to provide food on demand in space, but it should be noted that this novel approach to food production has many valuable terrestrial applications too. For example, enabling food production in climatically challenged environments including turning deserts into food bowls, or utilizing surplus electricity generated from large-scale renewable power sources.

Objective: To investigate the use of blood levels of N-(2 cyanoethyl)valine (CEVal), a haemoglobin adduct of acrylonitrile, to assess lack of compliance with smoking cessation. Methods: We report CEVal concentrations and percentiles over time for 2 cessation groups, compliance was established using NNAL concentrations. CEVal half-life was estimated. Optimal thresholds were calculated based on receiver operating characteristics analysis. Results: At 180 days, among 67 participants in the continued smoking group and 159 assigned to smoking cessation or sole THP use. CEVal half-life was estimated to be approximately 30 days and the optimal thresholds were for NNAL at 40 pg/mL and CEVal at 35 pmol/g globin (81% sensitivity and specificity). Conclusions: A new generation of biomarkers of compliance is required, specific to the new generation of nicotine products. Methodological validation and standardisation could allow robust assessment of effects across clinical and observational studies while promoting comparability between studies. CEVal could play an important role as biomarker of compliance for smoking cessation and switching studies.

Fenugreek seeds are widely used in Asia and other places of the world for their nutritive and medicinal properties. In Asia, Fenugreek seeds are also widely recommended for the geriatric populations. Here, we evaluated for the first time the effect of fenugreek seed feed supplementation on the liver antioxidant defense systems in aging mice. The study was conducted on 12 months aged mice which were given fenugreek seed dietary supplement. We have evaluated the activities of various antioxidant defense enzymes like superoxide dismutase (SOD), glutathione reductase (GR), and glutathione peroxidase (GPx), and estimated the phenolics and free radical scavenging properties in mice liver upon fenugreek supplementation. The estimation of SOD, GPx and GR activities in aged mice liver revealed a significant (P<0.01) difference among all the liver enzymes. Overall, this study reveals that fenugreek seed dietary supplementation has a positive effect in on the activities of the hepatic antioxidant defense enzymes in the aged mice.

The Mouse Grimace Scale (MGS) was developed 10 years ago to assess pain through characterisation of changes in five facial features or action units. The strength of the technique is that it is proposed to be a measure of spontaneous or non-evoked pain. A comprehensive scoping review of the academic literature was performed. The MGS has been employed mainly in evaluation of acute pain, particularly in the pain and neuroscience research fields. There has however been use of the technique in a wide range of fields, and based on limited study it does appear to have utility for pain assessment across a spectrum of animal models. Use of the method does allow detection of pain of a longer duration, up to a month post-initial insult. There has been less use of the technique using real-time methods and this is an area in need of further research.

Among other species, Black Soldier Fly (Hermetia illucens, HI, Diptera, Stratiomydae) has the great potential as food and feed ingredient in the EU, particularly thanks to its preference for organic waste as growth substrate. The production of insects as livestock feed or as food ingredient requires a strict monitoring of heavy metal content in the growth substrate in order to secure its safe. This study aims to investigate the presence of toxic metals cadmium, lead, mercury, arsenic, and nickel in HI prepupae and their growth substrates based on coffee roasting by-product and microlagae Schizochytrium sp. and Isochrysis sp. Analyses were carried out via graphite furnace atomic absorption spectrophotometry for Cd, Pb, Ni, and As, and via Direct Mercury Analyzer for Hg. All metal concentrations found in growth substrates were below the legal limit of undesirable substances in animal feed (2002/32/EC). Metals concentrations in HI prepupae were in the range (mg kg-1 wet weight): Cd 0.072–0.084, Pb 0.018–0.026, Hg 0.010–0.032, As 0.036–0.047, Ni 0.18–0.76. HI prepupae accumulate Cd, Pb and Hg, but our results indicate that the risk of exposure to metals from consumption of HI prepupae is relatively low and in compliance with European Union regulations.

Thousands of antibodies for diagnostic and other analytical purposes are on the market. However, it is often difficult to identify duplicates, reagent changes, and to assign the correct original publications to an antibody. This slows down scientific progress and might even be a cause of irreproducible research and a waste of resources. Recently, activities were started to suggest the sole use of recombinant antibodies in combination with the open communication of their sequence. In this case, such uncertainties should be eliminated. Unfortunately, this approach seems to be rather a long-term vision since the development and manufacturing of recombinant antibodies remain quite expensive in the foreseeable future. Also, nearly all commercial antibody suppliers may be reluctant to publish the sequence of their antibodies, since they fear counterfeiting. De-novo sequencing of antibodies is also not feasible today for a reagent user without access to the hybridoma clone. Nevertheless, it seems to be crucial for any scientist to have the opportunity to identify an antibody undoubtedly to guarantee the traceability of any research activity using antibodies from a third party as a tool. For this purpose, we developed a method for the identification of antibodies based on a MALDI-TOF-MS fingerprint. To circumvent lengthy denaturation, reduction, alkylation, and enzymatic digestion steps, the fragmentation was performed with a simple formic acid hydrolysis step. Eighty-nine unknown monoclonal antibodies were used for this study to examine the feasibility of this approach. Although the molecular assignment of peaks was rarely possible, antibodies could be easily recognized in a blinded test, simply from their mass-spectral fingerprint. A general protocol is given, which could be used without any optimization to generate fingerprints for a database. We want to propose that in most scientific projects relying critically on antibody reagents, such a fingerprint should be established to prove and document the identity of the used antibodies and to assign a specific reagent to a datasheet of a commercial supplier, a public database record or an antibody ID.

Current commercial, pre-commercial, and experimental in ovo techniques for the sex determination of fertilised eggs employ either minimally invasive biomolecular assays (extracting fluid via a small laser-drilled window in the eggshell, for detection of genetic or hormonal biomarkers), analysis of volatile compounds emitted from the eggshell, visible imaging, and reflectance or transmission spectroscopic analysis exploiting molecular optical fluorescence, polarisation, and scattering phenomena, including various combinations of these modalities. , to date no endeavour employing the NIR and FTIR based spectroscopic techniques has resulted in a commercially sustainable solution to the egg sexing problem. Besides achieving only subpar performance in overall accuracy, specificity, and sensitivity, the least invasive of the current state-of-the-art optical methods still requires, creating a transmission window (fenestration) of 12–15 mm diameter through to the mammillae layer of the shell, proximal to the external shell membrane, which can affect the incubation or post-hatch development viability of up to 10% of incubated eggs. Multimodal solution combining Raman spectroscopy and hyperspectral imaging has strong prospects to overcome the hard barriers existing before the perfection of a non-invasive in-line process for high reliability and rapid throughput for sex determination of eggs within 3 days of incubation. The method for sexing of chicken embryos needs to take a multipronged approach in collecting and analyzing spectral data that points to biomarkers using the machine learning approaches to look for nanomolar to picomolar concentrations of these in the fluid.

Many pigments and dyes are not only valuable molecules in manufacturing, but also pollutants. Science has certainly been producing sustainable alternatives to counter the issue of dyes, but this is not enough. There is also a need for change in the business attitude and leadership approach of the organizations that operate in the industry. Only through the successful combination of new technologies and forward-looking decision making, it will be possible to alter the status quo and deal with the multiple environmental and supply chain challenges that these businesses are and will be facing. With a focus on colorants, this critical review article will reflect on the importance of both a biotechnological approaches and sustainable leadership to achieve a sustainable, forward-looking fashion industry.

Recombinant laboratory plasmids (RLPs) are common in biological research and freely shared among academic research laboratories (ARLs), a practice required by many research funding agencies. However, the generation of accurate, reproducible results in experiments utilizing RLPs can be hampered by a lack of accompanying sequence information and metadata. This culture of RLP sharing without knowledge of sequence or etiology is accepted by publishers, not regulated by governments, and outside the realm of bio-industry. In addition, no centralized infrastructure currently exists to collate such data, which at the moment is fragmented across companies, non-profits, and governments and thus is not easily accessed or enacted toward threat assessment. The ubiquity, free exchange, and dual-use risk of RLPs exemplifies a biosecurity threat and elevates the need to characterize their composition to facilitate improved biorisk management by the academic community. A number of common sense solutions are available to create a culture that addresses the biosecurity gap posed by RLP sharing. Culture shift in RLP management will require new norms, effective data management for collation of RLP sequences and metadata, and an incentive structure that encourages sequencing by stakeholders. The next generation of researchers must initiate and champion this shift with support from funding agencies and endorsement from governments and international organizations. Coordination of efforts and stakeholders will require international public/private collaboration, a structure that will be critical to ensure widespread utility as well as the ability of lower-resourced partners to participate, contribute, and benefit.

The stress imposed by ethanol to Saccharomyces cerevisiae cells are one of the most challenging limiting factors in industrial fuel-ethanol production. Consequently, the toxicity and tolerance to high ethanol concentrations has been the subject of extensive research, allowing the identification of several genes important for increasing the tolerance to this stress factor. However, most studies were performed with well characterized laboratory strains, and how the results obtained with these strains work in industrial strains remains unknown. In the present work we have tested three different strategies known to increase ethanol tolerance by laboratory strains in an industrial fuel-ethanol producing strain: overexpression of the TRP1 or MSN2 genes, or overexpression of a truncated version of the MSN2 gene. Our results show that the industrial CAT-1 strain tolerates up to 14% ethanol, and indeed the three strategies increased its tolerance to ethanol. When these strains were subjected to fermentations with high sugar content and cell-recycle, simulating the industrial conditions used in Brazilian distilleries, only the strain with overexpression of the truncated MSN2 gene showed improved fermentation performance, allowing the production of 16% ethanol from 33% of total reducing sugars present in sugarcane molasses. Our results highlight the importance of testing genetic modifications in industrial yeast strains under industrial conditions in order to improve the production of industrial fuel ethanol by S. cerevisiae.

Aqueous and ethanolic extracts of Hibiscus sabdariffa were spray-drying using maltodextrin (MD) and gum arabic (GA) as carriers agents. An experimental design Taguchi L8 with seven variables was implemented. Physicochemical properties in the encapsulates were evaluated by UV-Vis, XRD, spectroscopy and gravimetric techniques. Treatments with aqueous extracts showed the highest concentration of total soluble polyphenols (TSP) 32.12- 21.23 mg EAG/g DW, and antioxidant capacity (AOX) for ABTS assay. The best treatment for TSP and AOX was T4: 2.5% Hibiscus w/w, aqueous extracts, decoction, extract-to-carrier ratio 1:1 (w/w), proportion to carriers (MD:GA) 80:20 (w/w), 10000 rpm, 150°C. Taguchi L8 model is a tool that allows the use of multiple variables with a low number of treatments that indicate the drying conditions that give the best parameters, focusing mainly on TSP and AOX, in addition, is a good alternative for the preservation and stability of the PC in Hibiscus.

Nanodiamonds of detonation origin are promising delivery agents of anti-cancer therapeutic compounds in a whole organism like mouse, owing to their versatile surface chemistry and ultra-small 5 nm average primary size compatible with natural elimination routes. However, to date, little is known about tissue distribution, elimination pathways and efficacy of nanodiamonds-based therapy in mice. In this report, we studied the capacity of cationic hydrogenated detonation nanodiamonds to carry active small interfering RNA (siRNA) in a mice model of Ewing sarcoma, a bone cancer of young adult due in the vast majority to the EWS-Fli1 junction oncogene. Replacing hydrogen gas by its radioactive analog tritium gas led to the formation of labeled nanodiamonds and allowed us to investigate their distribution throughout mouse organs and their excretion in urine and feces. We also demonstrated that siRNA directed against EWS-Fli1 inhibited this oncogene expression in tumor xenografted on mice. This work is a significant step to establish cationic hydrogenated detonation nanodiamond as an effective agent for in vivo delivery of active siRNA.

Manzamines are complex polycyclic marine-derived β-carboline alkaloids with reported anticancer, immunostimulatory, anti-inflammatory, antibacterial, antiviral, antimalarial, neuritogenic, hyperlipidemia and atherosclerosis suppression bioactivities, putatively associated with inhibition of glycogen synthase kinase-3, cyclin-dependent kinase 5, and vacuolar ATPases. We hypothesized that additional and yet undiscovered molecular targets might be associated with Manzamine A (MZA) reported pharmacological properties. We report herein for the first time to our knowledge that MZA inhibited a 90kDa ribosomal protein kinase S6 (RSK1) when screened against a panel of 30 protein kinases. Furthermore in vitro RSK kinase assays demonstrated a 10-fold selectivity in potency of MZA against RSK1 versus RSK2. MZA’s differential binding and selectivity toward the two isoforms is also supported by computational docking experiments. Specifically, the RSK1-MZA (N- and C-termini) complexes appear to have stronger interactions and preferable energetics contrary to the RSK2-MZA ones. In addition, our computational strategy suggests that MZA binds to the N-terminal kinase domain of RSK1 rather than the C-terminal domain. RSK is a vertebrate family of cytosolic serine-threonine kinases that act downstream of the ras-ERK1/2 (extracellular-signal-regulated kinase 1/2) pathway, which phosphorylates substrates shown to regulate several cellular processes including growth, survival and proliferation. Consequently, our findings have lead us to hypothesize that MZA and the 80 currently known manzamine-type alkaloids isolated from several sponge genera, may have novel pharmacological properties.

Microbe bioengineering has the potential to improve the nutritional value and health benefits of food products. Modifying microbial cells to produce specific proteins or compounds that can improve human health is one approach. Penicillin and recombinant insulin are two examples of this. The production of recombinant insulin by genetically modified E. coli bacteria has transformed diabetes treatment by providing a consistent and cost-effective source of insulin. Previously, insulin was extracted from the pancreas of animals, which was an expensive process that frequently resulted in impurities that could cause adverse reactions. Bioengineered insulin is now the standard diabetes treatment, providing a safe and effective method of controlling blood glucose levels. Bacteria, for example, can be genetically modified to produce vitamins like B12, which are difficult to obtain from plant sources. Likewise, yeasts can be genetically modified to produce β-carotene, a precursor to vitamin A that is required for vision, immune function, and skin health. Overall, bioengineered microorganisms have the potential to provide significant health benefits by producing essential compounds and proteins. Because of ongoing advances in genetic engineering techniques and understanding microbial metabolism, the possibilities for improving human health through bioengineering are limitless.

In continuation of our research on the influence of selenium incorporation on the biosynthesis, structure, immunomodulatory and antioxidant activities of polysaccharides of fungal origin, we have isolated from a post-culture medium of Lentinula edodes a selenium (Se)-containing exopolysaccharide fraction composed mainly of a highly branched 1-6-α-mannoprotein of molecular weight 4.5x106 Da, with 15% protein component. The structure of this fraction resembled mannoproteins isolated from yeast and other mushroom cultures, but it was characterized by a significantly higher molecular weight. X-ray absorption fine structure spectral analysis in the near edge region (XANES) suggested that selenium in the Se-exopolysaccharide structure was present mainly at the IV oxidation state. The simulation analysis in the EXAFS region suggested the presence of two oxygen atoms in the region surrounding the selenium. On the grounds of our previous studies, we hypothesized that selenium-enriched exopolysaccharides would possess higher biological activity than the non-Se-enriched reference fraction. To perform structure-activity studies, we conducted the same tests of biological activity as for previously obtained mycelial Se-polyglucans. The Se-enriched exopolysaccharide fraction significantly enhanced cell viability when incubated with normal (human umbilical vein endothelial cells, HUVEC) cells (but this effect was absent for malignant human cervical HeLa cells) and this fraction also protected the cells from oxidative stress conditions. The results of tests on the proliferation of human peripheral blood mononuclear cells suggested a selective immunosuppressive activity, like previously tested Se-polyglucans isolated from L. edodes mycelium. The Se-exopolysaccharide fraction, in concentrations of 10-100 µg/ml, inhibited human T lymphocyte proliferation induced by mitogens, without significant effects on B lymphocytes. As with previously obtained Se-polyglucans, in the currently tested Se-polymannans the selenium content increased the biological activity. However, the activity of selenium exopolysaccharides in all tests was significantly lower than that of previously tested mycelial isolates - most likely due to a different mode of selenium binding and its higher degree of oxidation.

This work aims to characterize a new method to recover low-manipulated human adipose tissue, enriched of adipose tissue-derived mesenchymal stem cells (ATD-MSCs) for autologous use in regenerative medicine applications. Lipoaspirated fat collected from patients was processed through Lipocell, a II-a medical device for dialysis of adipose tissue, by varying filter sizes and washing solutions. ATD-MSCs yield was measured with flow cytometry after SVF isolation in fresh and cultured samples. Purification from oil and blood was measured after centrifugation with spectrophotometer analysis. Extracellular matrix preservation was assessed through H&E staining and biochemical assay for total collagen, type-2 collagen, and GAGs quantification. Flow cytometry showed a 2-fold increase of ATD-MSCs yield in treated samples in comparison with untreated lipoaspirate; no differences where reported when varying filter size. The association of dialysis and washing thoroughly removed blood and oil from samples. Tissue architecture and extracellular matrix integrity were unaltered after Lipocell processing. Dialysis procedure associated with Ringer’s lactate preserves the proliferation ability of ATD-MSCs in cell culture. The characterization of the product shows that Lipocell is an efficient method to purify the tissue from undesired byproducts, preserving ATD-MSCs vitality and ECM integrity, resulting in a promising tool for regenerative medicine applications.

The integration of bioinformatics in drug discovery has revolutionized the field of pharmaceutical research. The use of computational tools and techniques has enabled researchers to analyze vast amounts of data, identify potential drug targets, and design new drugs with greater precision and efficiency. The integration of bioinformatics has also facilitated the development of personalized medicine, where drugs can be tailored to individual patients based on their genetic makeup. However, there are still challenges that need to be addressed, such as the need for more accurate predictive models and the ethical considerations surrounding the use of patient data. Overall, the integration of bioinformatics in drug discovery holds great promise for improving human health and advancing our understanding of disease mechanisms. In this mini-review, we discuss how Bioinformatics plays a crucial role in each step of drug discovery by providing tools and techniques to analyze large amounts of data generated from various sources, as well as the challenges and the opportunities offered by bioinformatics.

Shock waves, as used in medicine, can induce cell permeabilization, genetically transforming filamentous fungi; however, little is known on the interaction of shock waves with the cell wall. Because of this, the selection of parameters has been empirical. We studied the influence of shock waves on the germination of Aspergillus niger, to understand their effect on the modulation of four genes related to the growth of conidia. Parameters were varied in the range reported in protocols for genetic transformation. Vials containing conidia in suspension were exposed to either 50, 100 or 200 single-pulse or tandem shock waves, with different peak pressures (approximately 42, 66 and 83 MPa). In the tandem mode, three delays were tested. To equalize the total energy, the number of tandem “events” was halved compared to the number of single-pulse shock waves. Our results demonstrate that shock waves do not generate severe cellular effects on the viability and germination of A. niger conidia. Nevertheless, increase in the aggressiveness of the treatment induced a modification in the four genes tested. Scanning electron microscopy revealed significant changes to the cell wall of the conidia. Under optimized conditions, shock waves could be used for several biotechnological applications, surpassing conventional techniques.

: Microbiology has long been a keystone in fermentation and the utilization of yeast biology rein-forces molecular biotechnology as the pioneering frontier in brewing science. Consequently, modern understanding of the brewer’s yeast has faced significant refinement over the last few decades. This publication presents a condensed summation of Saccharomyces species dynamics with an emphasis on the relationship between traditional ale yeast, Saccharomyces cerevisiae, and the interspecific hybrids used in lager beer production, S. pastorianus. Introgression from other Sac-charomyces species is also touched on. The unique history of Saccharomyces cerevisiae and Saccharo-myces hybrids are exemplified by recent genomic sequencing studies aimed at categorizing brewing strains through phylogeny and redefining Saccharomyces species boundaries. Phylogenetic investigations highlight the genomic diversity of Saccharomyces cerevisiae ale strains long known to brewers by their fermentation characteristics and phenotypes. Discoveries of genomic contribu-tions from interspecific Saccharomyces species into the genome of S. cerevisiae strains is ever more apparent with increased investigations on the hybrid nature of modern industrial and historical fermentation yeast.

Mushrooms are unexploited treasures of secondary metabolites. Analysis of the chemical constituents of these mushrooms would be necessary for the assessment of their pharmacological and biological activities. This study aimed at profiling of mycochemical constituents of five wild mushroom extracts thereby understanding their biological and pharmacological properties. Mushrooms were collected from Arabuko-Sokoke and Kakamega National Reserved Forests, Kenya. Specimens were identified by both morphological and molecular methods. Bioactive compounds were extracted using chloroform, 70% ethanol, and hot water solvents. Chloroform, 70% ethanol, and hot water extracts of Auricularia auricula-judae, Microporus xanthopus, Termitomyces umkowaani, Trametes elegans, and Trametes versicolor were determined using gas chromatography and mass spectrometry (GC-MS). From all extracts, a total of fifty-one (51) compounds were identified and grouped into carboxylic acids, esters, phenols, fatty acids, alcohol, epoxides, aldehyde, fatty aldehyde, isoprenoid lipids, and steroid. Of the total compounds, Oleic acid (72.90%) from Trametes elegans was detected abundantly. Most of the compounds obtained from the chloroform extract of Trametes elegans and 70% ethanol extract of T. umkowaani are fatty acids. The identified compounds have revealed many biological and pharmacological activities such as antimicrobial, antioxidant, antimalarial, anti-inflammatory, insecticidal, anti-helminthic, larvicidal, vasodilator, antihypertensive, hepatoprotective, anticancer, antidiabetic, antifertility anti-diuretic, antiasthma, antifouling, anti-dermatophytic, antispasmodic, anti-hypocholesterolemic, nematicide, pesticide, immunostimulant, antiarthritic, and antihistaminic. These fatty acids are particularly playing important roles in the anti-inflammatory, hypocholesterolemic anticancer, and anti-biofilm formation activities. The presence of these bioactive components suggests that the extracts of five wild mushrooms could be good sources of secondary metabolites for drug discovery.

The integrity of the genome is crucial for the survival of all living organisms. However, genomes need to adapt to survive certain pressures and for this propose use several mechanisms to diversify. Chromosomal instability (CIN) is one of the main mechanism leading to the creation of genomic heterogeneity by altering the number of chromosomes as well as changing their structures. In this review we will discuss the different chromosomal patterns and changes observed in speciation, in evolutional biology as well as during tumor progression. By nature, human genome shows induction of diversity during gametogenesis but as well during tumorigenesis that can conclude in drastic changes such as whole genome doubling to more discrete changes as indels as well as the recent discovered complex chromosomal rearrangement chromothripsis. More importantly, changes observed during speciation are strikingly similar to the genomic evolution observed during tumor progression and resistance to therapy. The different origins of CIN will be treated as the importance of double strand breaks (DSB) or the consequences of micronuclei. We will also explain the mechanisms behind the controlled DSBs and recombination of homologous chromosomes observed during meiosis, to explain how errors lead to similar pattern observed during tumorigenesis. Then, we will also list several diseases associated to CIN resulting in fertility issue, miscarriage, genetic rare diseases and cancer. Understanding better chromosomal instability as a whole is primordial for the understanding of mechanisms leading to tumor progression.

Intergenerational Justice entitles the maximum retention of Earth’s Biodiversity. The United Nations COP 15 committed to the protection of 30% of Earth’s terrestrial environments to provide Biospheric Sustainability. However, proactive amphibian sustainability needs support from amphibian Reproduction Technologies, Biobanking, and Conservation Breeding Programs [RBCs]. These technologies include the reliable practice of stimulation of reproduction and the collection of sperm and oocytes, refrigerated and cryopreserved sperm storage, in vitro fertilization. Emerging technologies promise the perpetuation of species solely from biobanked biomaterials. However, although exemplary national programs for amphibian RBCs are now established in the USA and Australia, these potentials are not fully realised resulting in unacceptable biodiversity loss. To fully realise these potentials an independent and democratic global organisation is proposed to, 1) focus on the bioregions with the highest amphibian biodiversity, 2) maximise regional inclusion in international initiatives, and 3) provide for maximum community engagement and financial resources. We present the first overall multidisciplinary review of amphibian RBCs, including their geopolitical, cultural, and scientific background, along with their development and application to economically, efficiently, and reliably, perpetuate amphibian biodiversity. We provide genetic models, the potential for diploid cell cryopreservation to perpetuate species, along with significant new information on methodologies and techniques, and standardise nomenclature for specificity in publications and media consistency.

In the last two decades, extracellular vesicles (EVs) from the three domains of life, archae, bacteria and eukaryota, have gained increasing scientific attention. As such, the role of EVs in host-pathogen communication and immune modulation are being intensely investigated. Pivotal to EV research is the determination of how and where EVs are taken up by recipient cells and organs in vivo, which requires suitable tracking strategies including labelling. Labelling of EVs is often performed post-isolation which increases risks of non-specific labelling and the introduction of labelling artefacts. Here we exploited the inability of helminths to de novo synthesise fatty acids to enable labelling of EVs by whole organism uptake of fluorescent lipid analogues and the subsequent incorporation in EVs. We showed uptake of DOPE-Rhodamine in Anisakis spp. and Trichuris suis larvae. EVs isolated from supernatant of Anisakis spp. labelled with DOPE-Rhodamine were characterised to assess effects of labelling on size, structure and fluorescence of EVs. Fluorescent EVs were successfully taken up by the human macrophage cell line THP-1. This study therefore presents a novel staining method that can be utilized by the EV field in parasitology and potentially across multiple species.

Recent biotechnological innovations make feasible the new paradigm of creating biological neural circuits de novo. With advances in protein, cell and tissue engineering techniques, as well as cellular reprogramming methods, we are entering an era where the construction of neural circuits can open completely new ways for studying nervous systems and for treating nervous system disorders. I explore here three technologies, namely cellular engraftment, neuronal reprogramming and transsynaptic molecule engineering, and delineate how they are being used in a variety of basic research and translational medicine contexts. In basic neuroscience, neural circuit construction methods are enabling ways to study causality in neural development (e.g. neural precursor differentiation and migration) and circuit function (e.g. excitation/inhibition balance, neural population dynamics). In translational neuroscience, they are providing opportunities for the targeted correction of circuit malfunction in brain disorders, both psychiatric (e.g. schizophrenia) and neurological (e.g. Parkinson’s, Huntington’s and Alzheimer’s disease, as well as epilepsy). I discuss the challenges that these methods currently face, such as targeting specificity and cell survival, and outline future paths and opportunities to realize the full potential of technologies for creating new biological neural circuits.

We tested the hypothesis that concatemers of ancestral tRNAs gave rise to the 16S ribosomal RNA. We built an ancestral sequence of proto-tRNAs that showed a significant identity of 51.69% and a percentage of structural identity of 0.941 with the 16S ribosomal molecule. We also propose a hypothesis for the emergence of translation.

Improvement of agronomic traits in crops through gene editing (GE) relies on efficient trans-formation protocols for delivering the CRISPR/Cas9-coded transgenes. Recently, a few embryo-genesis-related genes have been described, the co-delivery of which significantly increases the transformation efficiency with reduced genotype dependency. Here we characterized the trans-genic and GE events in wheat (cv. Fielder) when transformed with the GROWTH-REGULATING FACTOR 4 (GRF4) and its cofactor GRF-INTERACTING FACTOR 1 (GIF1) chimeric gene. Trans-formation efficiency in our experiments ranged from 22% to 68%, and the editing events were faithfully propagated into the following generation. Both low and high copy number integration events were recovered in the T0 population with the various level of integrity of the left and right T-DNA borders. We also generated a population of wheat plants with ten different gRNAs targeting 30 loci in the genome. Comparison of the epigenetic profiles at the target sites and editing efficiency revealed a significant positive correlation between chromatin accessibility and muta-genesis rate. Overall, preliminary screening of the transgene quality and GE events in the T0 population of plants regenerated through the co-delivery of GRF-GIF can allow for the propagation of the best candidates for further phenotypic analysis.

A wide variety of bacteria, fungi and plants can produce bioactive secondary metabolites, which are often referred to as natural products. With the rapid development of DNA sequencing technology and bioinformatics, a large number of putative biosynthetic gene clusters have been reported. However, only a few natural products can be detected when isolated species are grown under conventional laboratory conditions, as most biosynthetic gene clusters are not expressed or are expressed at extremely low levels at these conditions. With the rapid development of synthetic biology, advanced genome mining and modification strategies have been reported, and they provide new opportunities for discovery of natural products. This review discusses advances in recent years that can accelerate the design, build, test, and learn (DBTL) cycle of natural product discovery, and prospects trends and key challenges for future research directions.

NH4Cl is one of the nitrogen sources for microalgal cultivation. However, excessive amounts of NH4Cl affects microalgal physiology and biomass contents. In this study, the effects of ammonium on microalgal growth and TAG content in the green microalga (Chlamydomonas reinhardtii) was investigated. Microalgal growth and TAG content under photoautotrophic conditions were found to be unchanged with 17 mM of ammonium, while this compound interfered with microalgal growth and induced TAG content under mixotrophic conditions with acetate supplementation. This suggested that ammonium could induce TAG production when acetate occurred in microalgal cultivation. Further, the effects of two different concentrations of NH4Cl (17 mM and 60 mM) on the cells under mixotrophic conditions were investigated. The results showed that both concentrations reduced microalgal growth, but induced total lipid and TAG content, especially after a 4-day cultivation. The oxygen evolution and Fv/Fm ratio showed that both concentrations completely inhibited the oxygen evolution on Day 4. The 60 mM NH4Cl reduced the Fv/Fm ratio from 0.7 to 0.48 indicating that ammonium supplementation directly affects the microalgae photosynthesis performance. A total of 1782 proteins were successfully identified using proteomics analysis. Among them, there were nine overexpressed proteins and four proteins were underexpressed. Using the protein–ligand interaction analysis, nitrogen metabolism is involved under NH4Cl conditions. This information can provide biochemical knowledge for microalgae development for sustainable energy usage.

Early development in mammals is characterized by the ability of each cell to produce a complete organism plus the extraembryonic, or placental, cells, defined as pluripotency. During subsequent development, pluripotency is lost, and cells begin to differentiate to a particular cell fate. This review summarizes the current knowledge of pluripotency features of bovine embryos cultured in vitro, focusing on the core of pluripotency genes (OCT4, NANOG, SOX2, and CDX2), and main chemical strategies for controlling pluripotent networks during early development. Finally, we will discuss the applicability of manipulating pluripotency during morula to blastocyst transition in cattle species.

Bacteriophages are viruses obligately infecting bacteria. They constitute the most numerous categories of biological forms populating our biosphere and are highly diverse and capable of infecting almost all bacteria. Phages make use of their host cell molecular machinery to express their own genes as they lack the ability to independently reproduce themselves. The inimitable characteristics of bacteriophages have enabled them to become propitious tools in biotechnology and genetic engineering. Phages show no tropism for mammalian cells but however, can be easily modified to present targeting ligands on their surface as coat protein fusions without any negative impacts on phage structure. These displayed ligands thereupon guide the recognition, interaction, and internalization of the phage into cells wherein efficiency of transfection is directly influenced by the copy number of the ligands used for targeting. Engineered phages are more efficacious for transgene delivery and gene expression in cancer cells when compared to other non-viral gene transfer strategies and are therefore being employed in developing cancer vaccines. The high level of stability as well as resistance of bacteriophages to various environmental conditions have enabled the development of virus-like particles (VLPs) capable of successful deliverance of several therapeutic drug cargos into tumors by selective targeting. Phage display technology has been used in therapy of Alzheimer’s disease and drug delivery into the brain. Exogenous peptides fused into the coat protein of phages enables the display of these peptides on the phage surface to generate combinatorial phage that facilitates their rapid separation using their ability to bind to a specific molecular target. Phage therapy has been shown to be safe in clinical settings when compared to antibiotics as it shows no adverse anaphylaxis nor adverse effects such as the emergence of multi-drug resistant bacteria. This review provides intriguing details of the use of natural and engineered phages in the therapy of diseases such as cancer, bacterial infections, bovine mastitis and dementia in addition to the use of CRISPR-Cas9 technology in generating genetically engineered phages. Further, the use of phage display technology in generating monoclonal antibodies against various human diseases is elucidated.

As a conceptual model of disease mechanisms, a disease map integrates available knowledge and is applied for data interpretation, predictions and hypothesis generation. It is possible to model disease mechanisms on different levels of granularity and adjust the approach to the goals of a particular project. This rich environment together with requirements for high-quality network reconstruction makes it challenging for new curators and groups to be quickly introduced to the development methods. In this review, we offer a step-by-step guide for developing a disease map within its mainstream pipeline that involves using the CellDesigner tool for creating and editing diagrams and the MINERVA Platform for online visualisation and exploration. We also describe how the Neo4j graph database environment can be used for managing and querying efficiently such a resource. For assessing the interoperability and reproducibility we apply FAIR principles.

This research aimed to establish the relationship between carbon-nitrogen nutritional factors and copper sulfate on laccase activity (LA) by Pleurotus ostreatus. Culture media composition was tested to choose the nitrogen source. Yeast extract (YE) was selected as a better nitrogen source than ammonium sulfate. Then, the effect of glucose and YE concentrations on biomass production and LA as response variables was evaluated using central composite experimental designs with and without copper. The results showed that the best culture medium composition was glucose 45 gL-1 and YE 15 gL-1, simultaneously optimizing these two response variables. The fungal transcriptome was obtained in this medium with or without copper, and the differentially expressed genes were found. Main up-regulated transcripts included three laccase genes (lacc2, lacc6, and lacc10) regulated by copper, whereas the principal down-regulated transcripts included a copper transporter (ctr1) and a regulator of nitrogen metabolism (nmr1). These results suggest that Ctr1, which facilitates the entry of copper in the cell, is regulated by nutrient-sufficiency conditions. Once inside, copper induces transcription of laccase genes. This finding could explain why a 10 to 20-fold increase in LA occurs with copper compared to cultures without copper when using the optimal concentration of YE as nitrogen sources.

Bee products have a millennial tradition in human use due to their numerous heal-ing properties. The paper presents the chemical characterization, antioxidant, anti-microbial and anti-inflammatory potential of royal jelly, apilarnil and propolis. These were obtained in the Banat area, Romania, in 2022. Royal jelly and apilarnil were analyzed in pure and lyophilized form and propolis in pure form and tincture. The percentage of moisture, dry matter, pH, acidity, content of impurities, sugars, proteins, ash, macro and micro minerals, total amino acids, polyphenols and flavo-noids were studied. The samples analyzed by us have a very well developed antiox-idant and according to the analysis of the mineral content of beekeeping products, an increased variability of macro and microminerals was determined, but also the lack of Pb in their composition. The flavonoid content of the samples was between 2.61519 mg QE/g sample for commercial pure apilarnil and 279.56115 mg QE/g sample for propolis harvested in 2021, and that of polyphenols between 0.63507 mg QE/g sample for natural royal jelly and 42.82186 mg QE/g sample in propolis with harvest in 2021. Propolis acts as a good antimicrobial and antiinflammatory agent, a fact that deserves to be researched in detail and exploited as a product with an ap-itherapeutic role for various microbial and inflammatory conditions.

Multinucleate cells can be produced in Dictyostelium by electric-pulse induced fusion. In these cells unilateral cleavage furrows are formed at spaces between areas that are controlled by aster microtubules. A peculiarity of unilateral cleavage furrows is their propensity to join laterally with other furrows into rings to form constrictions. This means, cytokinesis is biphasic in multinucleate cells, the final abscission of daughter cells being independent of the initial direction of furrow progression. Myosin-II and the actin-filament cross-linking protein cortexillin accumulate in the unilateral furrows, as they do in the normal cleavage furrows of mononucleate cells. Myosin-II is not essential for cytokinesis, but stabilizes and confines the position of the cleavage furrows.

Ganoderma boninense is the major causal agent for the basal stem rot (BSR) disease in oil palm, causing the progressive rot of the basal part of the stem. Despite its prominence, key pathogenicity determinants for the aggressive nature of hemibiotrophic infection remain unknown. In this study, genome sequencing and annotation of G. boninense T10 were carried out using the Illumina sequencing platform and comparative genome analysis was performed with previously reported G. boninense strains (NJ3 and G3). The pan-secretome of G. boninense was constructed and comprised of 937 core orthogroups, 243 accessory orthogroups, and 84 strain-specific orthogroups. A set of core candidate effector proteins (CEPs) were found to be enriched with catalytic protein classified as the carbohydrate-active enzymes, hydrolases as well as non-catalytic proteins. Differential expression analysis revealed an upregulation of CEP genes which was linked to the suppression of PTI signaling cascade while the downregulation of CEP genes was linked to the inhibition of PTI by preventing host defense elicitation. Genome architecture analysis revealed the one-speed architecture of the G. boninense genome and the lack of preferential association of CEP genes to the transposable elements. The findings obtained from this study would aid in the characterization of pathogenicity determinants and molecular biomarkers of BSR disease.

Abstract: The effect of collagen type 1 (Col I) oxidation on Adipose Tissue-Derived Mesenchymal Stem Cells (ADMSCs) remodeling is described as a model for acute oxidative stress. Morphologically, remodeling was presented by a mechanical rearrangement of adsorbed FITC-Col I and a trend for its organization in a fibril-like pattern - a process strongly abrogated in oxidized samples, but without visible changes in cell morphology. The cellular proteolytic activity was quantified in multiple samples utilizing fluorescence de-quenching (FRET effect). In the presence of ADMSCs a significant increase of native FITC-Col I fluorescence was observed, almost absent in the oxidized samples. Parallel studies in cell-free systems confirmed the enzymatic de-quenching of native FITC-Col I by Clostridial collagenase, again showing significant inhibition in oxidized samples. The structural changes in the oxidized Col I was further studied by Differential Scanning Calorimetry: an additional endotherm at 33.6°C along with the typical for native Col I at 40.5°C with sustained enthalpy (∆H) was observed in oxidized samples. Collectively, it has been evidenced that remodeling of Col I by ADMSCs is altered upon oxidation due to the intrinsic changes in the protein structure, thus presenting a novel mechanism for the control of stem cells' behavior toward collagen.

The brown seaweed Alaria esculenta is the second most cultivated species in Europe and it is therefore of interest to expand its application in developing food products. In this study, a lactic acid bacteria consortium (LAB consortium) consisting of three Lactiplantabacillus plantarum strains (relative abundance ~ 94%) and a minor amount of a Levilactobacillus brevis strain (relative abundance of ~ 6%) was investigated for its ability to ferment carbohydrates available in brown seaweed. The consortium demonstrated ability in fermenting glucose, mannitol, galactose, mannose, and xylose, of which glucose and mannitol were the most favored substrates; no growth was observed on fucose, mannuronic and guluronic acid. The consortium used different pathways for carbohydrate utilization and produced lactic acid as the main metabolite. In glucose fermentations, only lactic acid was produced, but use of mannitol as carbohydrate source, resulted in co-production of lactic acid, ethanol and succinate. Xylose fermentation resulted in acetate production. The consortium was also able to utilize laminari-oligosaccharides (DP 2-4), obtained after enzymatic hydrolysis of laminarin, and produced lactic acid as metabolite. The consortium could grow directly on A. esculenta, resulting in a pH decrease to 3.8 after 7 days of fermentation. Incubation of the same seaweed at corresponding conditions without inoculation resulted in spoilage of the seaweed by endogenous bacteria.