The aim of this study was to examine pathologic complete response (pCR) and overall survival (OS) of patients diagnosed with non-metastatic inflammatory breast cancer (IBC). A total of N=8,550 cases undergoing surgery were identified between 2004-2013, using the National Cancer Database (NCDB). Patients were grouped into 4 biologic subtypes (HR+/HER2-, HR+/HER2+, HR-/HER2+, HR-/HER2-). The median age at diagnosis was 56 years. On average, women were followed for 3.7 years [interquartile range=3.0]. The majority were white (80%), had private health insurance (50%), and presented with poorly differentiated tumors (57%). Approximately 46% of the cancers were >5cm. Most patients underwent mastectomy (94%) and received radiotherapy (71%). Differences by biologic subtypes were observed for grade, lymph node invasion, race, and tumor size (p<.0001). Compared with non-pCR (54%), patients experiencing pCR had superior 5-year survival (77%) (p<.0001). Survival was poor for triple-negative (TN) tumors (37%) vs. other biologic subtypes (60%) (p<.0001). On multivariable analysis, TN-IBC, positive margins, and not receiving either chemotherapy, hormonal therapy or radiotherapy were independently associated with poor 5-year survival (p<.0001). In this large multicentric analysis of IBC, categorized by biologic subtypes, we observed significant differential tumor, patient and treatment characteristics, and OS.

Retention of placenta (RP) in crossbred cattle poses health, reproductive, and economic challenges linked to oxidative stress, hormonal imbalances, and complex causative factors. Emphasizing the key role of mitochondria in oxidative phosphorylation (OXPHOS) and hormone synthesis, our study is the first to explore differences in OXPHOS activity, mitochondrial content, and mtDNA-encoded gene expression between normal and RP. In addition, gross morphological examination of placental tissues from both groups revealed noticeable disparities in cotyledons of RP as compared to normal placenta. Moreover, OXPHOS complex I specific activity was decreased in RP, and RT-qPCR analysis unveiled a significant reduction in transcript levels of 10 mtDNA-encoded protein genes. While COX3 and ATP8 genes showed no significant changes, ND6 increased in RP. Whereas, mtDNA copy number varied between animals but showed no statistically significant differences among groups. Retained placenta manifests alterations in the transcript levels of various mitochondrial genes, complex activity, and mtDNA content, potentially linked to heightened energy demands or mitochondrial insufficiency, posing an elevated risk. Overall, our study suggests a link between mitochondrial abnormalities, gene expression, and RP. This connection could influence the removal of fetal membranes by affecting energy supply, hormonal balance, and oxidative stress. This study also offers novel insights and serves as a valuable animal model for studying placental insufficiency in humans. Moving forward, comprehensive investigations are imperative to unravel underlying mechanisms and intricate roles of mitochondria in the context of RP, paving the way for targeted interventions to mitigate health, reproductive, and economic challenges in crossbred cattle.

Potato virus Y (PVY) is one of the most important constraints to potato production worldwide. There is an increasing occurrence of recombinant PVY strains PVYNTN and PVYN-Wi and a decline in the incidence of the nonrecombinant PVYO. We hypothesized that this may be due to the ability of these recombinant strains to antagonize and/or outcompete PVYO in mixed infections. To deter-mine this, we investigated interactions between PVYO and three recombinant PVY strains com-mon in North America: PVYNTN, PVYN-Wi, and PVYN:O. Overall, our study showed that these inter-actions are tissue dependent. Specifically, PVYNTN, the main causal agent of potato tuber necrotic ringspot disease (PTNRD), was found to be more adaptable than PVYO, especially in potato leaves due, at least in part, to the Ny gene that confers hypersensitive resistance (HR) to PVYO. Further-more, PVYN-Wi was found to repress PVYO in potato tubers but act synergistically in potato leaves. The PVYO-induced foliage necrosis in cultivar ‘Ranger Russet’ was observed to be more severe in plants co-infected by PVYN-Wi and PVYN:O, respectively, resulting in plant death. Strikingly, this PVYO -induced necrosis was suppressed by PVYNTN in doubly infected plants. These interactions may, at least partially, explain the decreasing incidence of PVYO in United States potato produc-tion regions, especially given that many cultivars contain the Ny gene, which likely limits PVYO enabling PVYNTN and PVYN-Wi to outcompete. We also found that replication and cell-to-cell movement of these PVY strains in tubers at 4C was similar to levels at ambient temperature.

We present atomic structures and nonequilbrium synthesis of new class of materials, where the basic structural unit is a diamond tetrahedron. When units of one, two, and three tetrahedra are randomly packed, we create distinct phases of amorphous Q-carbon. Four tetrahedra in two adjacent layers lead to crystalline diamond lattice, which has four missing tetrahedra alternately. When these four missing tetrahedra are filled, we create subunit cell of crystalline Q-diamond. Theoretical calculations show that superconducting transition temperature (Tc) in 50 atomic % B-doped Q-diamond can reach near room temperature at ambient pressures. This is consistent with our earlier results using low-loss EELS measurements in 50 atomic % B-doped Q-carbon, which had mostly amorphous QB3 phase mixed with some crystalline Q-diamond phase. These EELS results showed that the Tc for these samples was in between 90K and 300K. Theoretical calculations of density of states, Eliashberg function, electron-phonon interaction parameter, and root-mean-square and logarithmic average of frequency in crystalline Q-diamond show Tc in the range of 268K to 300K, which is in a complete agreement with our EELS results in QB3.

The risk of the emergence and reemergence of zoonoses is high in regions that are under anthropogenic actions, as they contribute to the risk of vector disease transmission. Yellow fever (YF) is among the main pathogenic arboviral disease in the world, and the Culicidae Aedes albopictus has been proposed to have the potential to transmit yellow fever virus (YFV). This mosquito inhabits both urban and wild environments, and under experimental conditions, it has been shown to be susceptible to infection by YFV. In this study, the vector competence of the mosquito Ae. albopictus for the YFV was investigated. Female Ae. albopictus were exposed to non-human primates (NHP) of the genus Callithrix infected with YFV for blood meal. Subsequently, on the 14th and 21st days post infection, the legs, heads, thorax/abdomen and saliva of the arthropods were collected and analyzed by viral isolation and molecular analysis techniques to verify the infection, dissemination and transmission. The presence of YFV was detected in saliva samples through viral isolation and in the head, thorax/abdomen and legs both by viral isolation and by molecular detection. The susceptibility of Ae. Albopictus to YFV confers a potential risk of reemergence of urban YF in Brazil.

Since the UK commenced newborn screening for isovaleric acidemia in 2015, changes in prescribing have increased the incidence of false positive (FP) results due to pivaloylcarnitine. A review of screening results between 2015-22 identified 24 true positive (TP) and 84 FP cases, with pivalate interference confirmed in 76/84. Initial C5 carnitine (C5C) did not discriminate between FP and TP with median (range) C5C of 2.9 (2.0 - 9.6) and 4.0 (1.8 - &gt;70) µmol/L respectively and neither did Precision Newborn Screening via Collaborative Laboratory Integrated Reports (CLIR), which identified only 1/47 FP cases. However, among the TP cases, disease severity showed a correlation with initial C5C, in ‘asymptomatic’ individuals (n=17), demonstrating a median (range) C5C of 3.0 (1.8 – 7.1) whilst ‘clinically affected’ patients (n=7), showed a median (range) C5C of 13.9 (7.7 - &gt; 70) µmol/L. These findings allowed the introduction of dual cut-off values into the screening algorithm to reduce the incidence of FPs, with initial C5C results ≥ 5 µmol/L triggering urgent referral, and those &gt; 2.0 and &lt; 5.0 µmol/L prompting 2nd tier C5-isobar testing. This will avoid delayed referral in babies at particular risk whilst reducing the FP rate for the remainder.

The aim of the study was to determine the occurrence and characteristics of coagulase-positive Staphylococcus strains in the carcasses of wild birds and aviary capercaillies in south-eastern Poland. Samples from capercaillies and wild birds belonging to 12 orders and 26 species were tested. The resistance of the strains and the presence of selected virulence genes were examined. Similarity analysis of the isolates was performed using pulsed-field gel electrophoresis (PFGE). Coagulase-positive Staphylococcus strains accounted for 5.7% and belonged to the species: Staphylococcus aureus, Staphylococcus pseudintermedius and Staphylococcus delphini. Among isolated strains, 15.8% were multidrug resistant. The most frequently detected virulence genes were hla in 58% of isolates and hlb and hld in 47.4% of isolates., The results of multiplex PCR showed the presence of genes responsible for the production of enterotoxins C, B, E, and J, in single isolates. It can be concluded that coagulase-positive Staphylococcus strains accounted for a small percentage of staphylococci isolated from free-living birds in the study area. The occurrence of multidrug-resistant coagulase-positive Staphylococcus strains in aviary capercaillies suggests that they play a role in the transmission and spread of resistant strains into the environment. Free-living birds may be also a significant reservoir of enterotoxigenic Staphylococcus strains.

The pursuit of sustainable wellbeing is one of the research objectives of positive experience design. Driven by this goal, the purpose of this paper is to provide an impact-centered sustainable positive experience design model. Firstly, the literature review method was used to define the research status and concept of impact-centered sustainable design. Secondly, an impact-centered sustainable positive experience design model was constructed, and relevant formulas for concept generation and concept evaluation were proposed. Thirdly, design verification was conducted through a workshop. Finally, the Technology Acceptance Model (TAM) questionnaire was used to evaluate and discuss the design model. An impact-centered sustainable positive design model was proposed, which includes the important impact dimensions of sensory experience and meaningful experience on users’ quality of life at different levels: healthy living (pleasure index, health behavior), harmonious community (social connectivity, social contribution), and livable environment (living environment, environmental contribution). Based on positive experience related theory, this study takes long-term impacts as the starting point for sustainable positive experience design, which helps designers to generate design concepts from a systematic and long-term perspective.

The current study aimed to examine the relationship between demographic variables within the family context and parenting behaviors among adolescent mothers (including affection, responsiveness, encouragement, and teaching). These factors were correlated with communication, problem-solving abilities, and personal-social development in typically developing infants. The study included a sample of 79 Chilean adolescent mother-child dyads with children aged 10 to 24 months. Communication, problem-solving, and personal-social development were assessed using the Ages and Stages Questionnaire-3, along with a demographic information questionnaire. Parenting behaviors (affection, responsiveness, encouragement, and teaching) were observed using the Spanish version of Parenting Interactions with Children: Checklist of Observations Linked to Outcomes. The findings indicated that employed mothers and those who had not dropped out of school had children with better problem-solving skills. Additionally, children residing with their fathers and female children exhibited superior performance in communication, problem-solving, and personal-social development. Maternal responsiveness was associated with communication and problem-solving, while maternal encouragement was linked to improved problem-solving skills. Maternal teaching was connected to communication, problem-solving, and personal-social development. The study emphasized the significance of parenting and sociodemographic factors among adolescent mothers in influencing their children's development.

Positive recurrence for a single-server queueing system is established under generalised intensity conditions of service which does not assume existence of the density distribution function of service but a certain integral type lower bound as a sufficient condition. Positive recurrence implies existence of the invariant distribution and a guaranteed slow convergence to it in the total variation metric.

Secure attachment, developed through consistent relationships with attachment figures in childhood, is a crucial factor in fostering healthy interpersonal relationships and a positive self-perception. This research aimed to assess how perceived security, as an indicator of secure attachment, predicts adult self-concept through the mediating effects of resilience and positive self-esteem. A quantitative, cross-sectional, and non-probabilistic study was conducted with 383 participants, of whom 76.2% were females, aged between 17 and 86 years (M = 28.52, SD = 12.23). Correlational analyses revealed significant positive associations between perceived security, self-concept, resilience, and positive self-esteem. Path analysis demonstrated that resilience and positive self-esteem sequentially mediate the relationship between perceived security during childhood and adult self-concept (Total Effect R2 = .34, F(1,381) = 193, p < .001; Indirect effect of sequential mediation b = .02, CI 95% [.01, .04]). Findings suggest that the ability to cope with ad-versity and personal acceptance underlie the impact of perceived security on self-concept. It is recommended to promote not only secure attachment in family interventions but also resilience and self-esteem in programs targeting adolescents, youth, and adults.

We study the existence of positive solutions for a system of Hadamard fractional differential equations on an infinite interval with nonnegative nonlinearities, subject to coupled nonlocal boundary conditions which contain Hadamard fractional derivatives and Riemann-Stieltjes integrals. In the proof of the main results, we use the Guo-Krasnosel'skii fixed point theorem and the Leggett-Williams fixed point theorem.

A The aim of this article is to present the results of studies investigating the relationship between trust in supervisor and in co-workers and trust in the organization as an institution on the one hand, and employee well-being on the other hand. The presented research project was carried out from the perspective of building a positive organization with employee well-being as one of the major goals. The research covered 1,113 individuals, employed with various organizations across Poland. The sample consisted of 48% women and 52% men. Trust was measured using the Trust in Supervisor Scale, Affect- and Cognition-Based Trust, concerning trust in co-workers, and the original Trust in the Workplace Questionnaire. The Eudemonic Well-Being in the Workplace Questionnaire was used to measure well-being. Canonical correlation analysis was used to assess simultaneous associations between sets of measured variables. The results prove that trust, including trust in co-workers, in supervisor and in the organization as an institution, promotes employee well-being in all its dimensions.

The therapeutic effects of volatile anesthetics on mental diseases, particularly schizophrenia, have gained considerable interest. Although isoflurane is a commonly used volatile anesthetic, there’s no more evidence that it could work on treating schizophrenia. Here, we discovered that inhaling isoflurane at low concentrations might reverse the behavioral phenotypes of schizophrenia caused by MK801, such as hyperlocomotion, pre-pulse inhibition impairment, and working memory loss. Isoflurane also helped recovering adult neurogenesis and synaptic plasticity impairments in the dentate gyrus (DG) induced by MK801. To better understand the mechanism, we discovered that isoflurane could reverse the reduction of parvalbumin (PV)-positive GABAergic interneuron (PVI) number and the aberration of NRG1-ErbB4 signaling in the DG; however, isoflurane could not reverse the schizophrenia-related phenotypes caused by PVI ablation, indicating that PVI are necessary for the therapeutic effect of isoflurane. Interestingly, isoflurane could reverse phenotypes caused by blocking PVIs GABA release in the DG, indicating the therapeutic impact is independent of PVI GABA release. Our research revealed that isoflurane might be used to treat schizophrenia, possibly through PVI in the DG.

The purpose of this review is to present the current knowledge about the diagnostic and treatment options in bladder cancer (BCa) patients with clinically positive lymph nodes (cN+). In this review compaction of CT and MRI performance in preoperative prediction of lymph node invasion (LNI) in BCa patients was presented, along with other diagnostic methods. Most scientific societies do not distinguish cN+ patients in their guidelines, recommendations concern muscle-invasive bladder cancer (MIBC) and differ between associations. Currently, the standard treatment of patients with MIBC is radical cystectomy (RC) with bilateral pelvic lymph node dis-section (PLND). The template of PLND and its therapeutic value remain debatable. Moreover, most guidelines recommend neoadjuvant chemotherapy (NAC). However, there is still lack of definitive evidence of the superiority of neoadjuvant chemotherapy over adjuvant chemotherapy. Nevertheless, the curative treatment that provides the best long-term survival in cN+ patients is a multimodal approach with a combination of chemotherapy and RC. Recent studies demonstrate the growing importance of immunotherapy. Special attention should be paid to cN+ BCa patients as the oncological outcomes are significantly worse for this group.

The needs for environmental reporting to include positive outcomes considering differences between creation of less harm, benefits and net benefits are explored. To become mainstream, nature-positive development needs positive messaging, measures and metrics to guide, plan and assess urban outcomes. With the accelerating climate crisis and negative messages getting the upper-hand, it’s important to avoid paralysis by bad news. Whilst striving for a nature-positive world, more effort should be on moving beyond zero to qualify and quantify benefits, gains, and regenerative outcomes instead of oscillating around damage and loss sticking points. Life Cycle Benefit Assessment (LCBA) is a method to measure gains in accelerating restoration and climate security. It enables a good news focus as its reach is to quantify and show positive gains beyond the negative and zero loss outcomes. The paper aims to clarify concepts, challenges and quantitative methods then review real-world third-party-certified case studies. Climate security, human wellness and resource viability gains inside safe operating space within planetary boundaries are quantified as positive benefits. Contrary to conventional Life Cycle Impact Assessment (LCIA) LCBA assigns damage and loss as negative debts and benefit as positive gains. It concludes that LCBA offers business and design a new environment assessment tool, with research needed on economic and other outcomes.

Schizophrenia comprises various symptom domains, including positive and negative symptoms. Machine learning showed that a) negative symptoms are significantly interrelated with PHEM (psychosis, hostility, excitation, and mannerism) symptoms, formal thought disorders (FTD) and psychomotor retardation (PMR); and b) stable phase schizophrenia comprises two distinct classes, namely Major Neuro-Cognitive Psychosis (MNP, largely overlapping with deficit schizophrenia) and Simple NP (SNP). In this study, we recruited 120 MNP patients and 54 healthy subjects and measured the above-mentioned symptom domains. In MNP, there were significant associations between negative and PHEM symptoms, FTD and PMR. A single latent trait, which is essentially unidimensional, underlies these key domains of schizophrenia and MNP and additionally shows excellent internal consistency reliability, convergent validity, and predictive relevance. Confirmatory Tedrad Analysis indicates that this latent vector fits a reflective model. The lack of discriminant validity shows that PHEM and negative symptoms greatly overlap and probably measure the same construct. Soft Independent Modeling of Class Analogy (SIMCA) shows that MNP (diagnosis based on negative symptoms) is better modeled using PHEM symptoms, FTD, and PMR than negative symptoms. In conclusion, in stable phase MNP, a restricted sample of the schizophrenia population, negative and PHEM symptoms, FTD and PMR belong to one underlying latent vector reflecting overall severity of schizophrenia (OSOS). The bi-dimensional concept of “positive” and “negative” symptoms cannot be validated and, therefore, future research in stable phase schizophrenia should consider that the latent phenomenon OSOS as well as its 8 reflective manifestations are the key factors of schizophrenia phenomenology.

Apple red flesh coloration is a result of the performance of biochemical pathway involved in the biosynthesis of anthocyanins and anthocyanidins. Based on apple genome analysis, high number of regulatory genes, mainly transcription factors such as MYB, which are components of regulatory complex MYB-bHLH-WD40, and several structural genes (PAL, 4CL, CHS, CHI, F3H, DFR, ANS, UFGT) involved in anthocyanin biosynthesis have been identified so far. In this study, we investigated previously unrecognized genes involved in the constitution of red-flesh apple phenotype. These genes could be concerned as molecular markers for early selection of new apple cultivars. Based on comparative transcriptome analysis of apples with different fruit flesh coloration, we successfully identified and characterized ten genes from plant hormone transduction pathway of: auxin (GH3); cytokinins (B-ARR); gibberellins (DELLA); abscisic acid (SnRK2 and ABF); brassinosteroids (BRI1, BZR1 and TCH4); jasmonic acid (MYC2); and salicylic acid (NPR1). In presented studies an analysis of their expression profiles was performed in immature and ripe fruits of red-fleshed cultivars. Generally, we have uncovered, the set of the novel genes, mediating regulation of the abscisic acid, salicylic acid, cytokinins and jasmonic acid signaling and preliminarily described their role in anthocyanin biosynthesis, accumulation and degradation. Their expression was not characterized in apple fruits so far. Presented study, allowed to underline the relationship of genes from hormone signal transduction pathway with UFGT gene, directly responsible for anthocyanin color transformation as well as confirmed their crucial role of plant hormone regulation and anthocyanin accumulation during apple fruit ripening.

Gastric cancer (GC) is a highly heterogeneous, complex disease and the fifth most common cancer worldwide (about one million cases and 784 000 deaths worldwide in 2018). CG is lately diagnosed and guarantees a poor prognosis for GC (the 5-year survival rate is less than 20%, but in early detection can reach 90%). This study evaluated the transcriptional profile in tumor gastric samples to find genes highly expressed during tumor establishment and use the related proteins as targets to find new anticancer molecules. Data was collected at Gene Expression Omnibus (GEO) bank to obtain 3 dataset matrices that analyze gastric tumor tissue versus normal gastric tissue, performed microarray using GPL570 platform, and from different sources. The genes found in silico analysis were confirmed in several lines of GC cells by RT-qPCR. The protein data bank was used to find the correspondent proteins. Then a structural-based virtual screening was done to find molecules, and docking analysis was done using the DockThor server. Our results of transcriptomic analysis, together with RT-qPCR, confirm the high expression of the genes AJUBA, FBXL13, CCDC69, CD80 and NOLC1 in GC lines. Based on that, the correspondent proteins were used in SBVS analysis. Five molecules, one each target, MCULE-2386589557-0-6, MCULE-7343047040-0-1, MCULE-5230409338-0-3, MCULE-9178344200-0-1 and MCULE -5881513100-0-29. All molecules have favorable pharmacokinetic, pharmacodynamic and toxicological properties. Molecular docking analysis revealed that the molecules interact with proteins in critical sites for their activity. Using a virtual screening approach, a molecular docking study was performed for proteins encoded by genes that play important roles in cellular functions for carcinogenesis. Combining a systematic collection of public microarray data with a comparative meta-profiling, RT-qPCR, SBVS, and molecular docking analysis provided a suitable approach to find genes involved in GC and work the correspondent protein to strive for new molecules with anticancer properties.

The coronavirus disease 2019 (COVID-19) pandemic has led to wastewater surveillance becoming an important tool for monitoring the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) within communities. As a result, molecular methods, in particular reverse transcription-quantitative PCR (RT-qPCR), have been employed to generate large data sets aimed at the detection and quantification of SARS-CoV-2 in wastewater. Although RT-qPCR is rapid and sensitive, there is no standard method that fits all use cases, there are no certified quantification standards and experiments are carried out using numerous different assays, reagents, instruments, and data analysis protocols. These variations can lead to the reporting of erroneous quantitative data resulting in potentially misleading interpretations and conclusions. We have reviewed the SARS-CoV-2 wastewater surveillance literature focusing on the variability of RT-qPCR data as revealed by inconsistent standard curves and associated parameters. We find that variation in these parameters and deviations from best practices as described in The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines suggest a lack of reproducibility and reliability in quantitative measurements of SARS-CoV-2 RNA in wastewater.

Coronavirus-infected diseases have posed great threats to human health. In past years, highly infectious coronavirus-induced diseases, including COVID-19, SARS, and MERS, have resulted in world-wide severe infections. Our literature annotations identified 110 chemical drugs and 26 antibodies effective against at least one human coronavirus infection in vitro or in vivo. Many of these drugs inhibit viral entry to cells and viral replication inside cells or modulate host immune responses. Many antimicrobial drugs, including antimalarial (e.g., chloroquine and mefloquine) and antifungal (e.g., terconazole and rapamycin) drugs as well as antibiotics (e.g., teicoplanin and azithromycin) were associated with anti-coronavirus activity. A few drugs, including remdesivir, chloroquine, favipiravir, and tocilizumab, have already been reported to be effective against SARS-CoV-2 infection in vitro or in vivo. After mapping our identified drugs to three ontologies ChEBI, NDF-RT, and DrON, many features such as roles and mechanisms of action (MoAs) of these drugs were identified and categorized. For example, out of 57 drugs with MoA annotations in NDF-RT, 47 have MoAs of different types of inhibitors and antagonists. A total of 29 anticoronaviral drugs are anticancer drugs with the antineoplastic role. Two clustering analyses, one based on ChEBI-based semantic similarity, the other based on drug chemical similarity, were performed to cluster 110 drugs to new categories. Moreover, differences in physicochemical properties among the drugs were found between those inhibiting viral entry and viral replication. A total of 163 host genes were identified as the known targets of 68 anti-coronavirus drugs, resulting in a network of 428 interactions among these drugs and targets. Chlorpromazine, dasatinib, and anisomycin are the hubs of the drug-target network with the highest number of connected target proteins. Many enriched pathways such as calcium signaling and neuroactive ligand-receptor interaction pathways were identified. These findings may be used to facilitate drug repurposing against COVID-19.

Achieving effective control over microbial contamination necessitates the precise and concurrent identification of numerous pathogens. In this research, we have devised a remarkably sensitive duplex droplet digital PCR (dddPCR) reaction system to simultaneously detect Pseudomonas aeruginosa (P. aeruginosa) and Pseudomonas fragi (P. fragi). Employing comparative genomics, we identified four genes of P. fragi. By specific analysis, RS22680 gene was selected as the detection target of P. fragi and LasR gene was chosed as P. aeruginosa, which were applied to construct a dddPCR reaction. In terms of specificity, sensitivity and anti-interference ability, the constructed dddPCR detection system was verified and analyzed. The assay showed excellent sensitivity and applicability, as evidenced by a limit of detection of 100 CFU/mL. When the concentration of natural background bacteria in milk or fresh meat was 100 times that of the target detection bacteria, the method was still capable of completing the absolute quantification. In the simulation of actual sample contamination, P. aeruginosa could be detected after 3 h of enrichment culture, and P. fragi could be detected after 6 h. The established ddPCR detection system exhibits exceptional performance, serving as a foundation for the simultaneous detection of various pathogenic bacteria in food products.

Gas gangrene refers to severe necrotizing syndromes in soft tissues of animals caused by Clos-tridium. Five species of histotoxic clostridia are responsible for myonecroses. These agents require a port of entry, such as skin punctures with needles contaminated with bacterial spores. The present case report describes the clinicopathological findings of gas gangrene in a male Quarter horse living in the Brazilian Amazon biome. Clinical examination revealed clinical signs of toxemia and an enlarged left limb. The anamnesis found the administration of medication to this limb. The animal died five hours after its arrival at the veterinary hospital. At necropsy, histopathological lesions characterized by intense muscle necrosis, liquefaction, and gas formation were observed, suggestive of Clostridium lesions, which were confirmed by multiplex PCR. C. septicum was the etiological agent identified.

The Herpotrichiellaceae family is an important group of dematiaceous filamentous fungi, associated with a variety of pathogenic fungal species causing Chromoblastomycosis (CBM) and Phaeohyphomycosis (PHM), both with polymorphic clinical manifestations and worldwide incidence. Currently, the identification of this family and determination of the causative agent is challenging due to the subjectivity of morphological identification methods, necessitating the use of molecular techniques to complement diagnosis. In this context, genetic sequencing of the Internal Transcribed Spacer (ITS) has become the norm due to a lack of alternative molecular tools for identifying these agents. Therefore, this study aimed to develop PCR-Multiplex methodologies to address this gap. Sequences from the ITS and Large subunit (LSU) of ribosomal DNA were used, and after manual curation and in vitro analyses, primers were synthesized for the identification of the aforementioned targets. The primers were optimized and validated in vitro, resulting in two PCR-Multiplex methodologies: one for identifying the Herpotrichiellaceae family and the bantiana clade, and another for determining the species Fonsecaea pedrosoi and Fonsecaea monophora. Ultimately, the assays developed in this study aim to complement other identification approaches for these agents, reducing the need for sequencing, improving the management of these infections, and enhancing the accuracy of epidemiological information.

The genus Colias Fabricius, 1807 includes numerous taxa and forms with uncertain status and taxonomic position. Among such taxa are Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897, interpreted in the literature either as conspecific forms, as subspecies of different but morphologically somewhat similar Colias species, or as distinct species-level taxa. Based on mitochondrial and nuclear DNA markers, we reconstruct a phylogeographic pattern of the taxa in question. We recover and include in our analysis DNA barcodes of the century-old type specimens, the lectotype of C. tamerlana deposited in the Natural History Museum (Museum für Naturkunde), Berlin, Germany (ZMHU), the paralectotype of C. tamerlana and the lectoptype of C. mongola deposited in the Zoological Institute, Russian Academy of Sciences, Saint-Petersburg, Russia (ZISP). Our analysis grouped all specimens within four (HP_I-HP-IV) deeply divergent but geographically poorly structured clades which did not support nonconspecifity of C. mongola–C. tamerlana. We also show that all studied females of the widely distributed haplogroup HP_II were infected with a single Wolbachia strain belonging to the supergroup B, while the males of this haplogroup, as well as all other investigated specimens, were not infected. Our data underline the relevance of large-scale sampling dataset analysis and the need of testing for Wolbachia infection to avoid erroneous phylogenetic reconstructions and species misidentification.

Honey bees are vital organisms that provide ecological and economic value to humans. However, in recent years, the increase in honey bee losses due to various environmental factors, including pesticides, has become a growing concern. In Europe, neonicotinoid pesticides that are banned are being used without restrictions in the domestic setting, leading to ongoing damages as a result. Ongoing research is continually being conducted to demonstrate the risks associated with neonicotinoid pesticides. However, validation of the actual damages and impact in the field remains absent. Therefore, in this study, we observed changes in honey bee (Apis mellifera) colonies located near rice cultivation areas as they progressed beyond the rice pesticide application period. Furthermore, we collected honey bees exposed to the clothianidin and analyzed their stress-related gene expression. The results showed that the foraging behavior of honey bee colonies located near rice cultivation areas did not exhibit significant differences between the treatment site (Cheongyang and Gimje) and the control site (Wanju) during the experimental period. However, it was observed that the expression levels of stress-related genes in honey bees collected from the treatment group were significantly higher than those in the control. Most of the stress-related genes were associated with detoxification processes in response to pesticides. As a result, pesticide treatment in proximity to rice cultivation areas did not cause direct damage to honey bees but had an indirect impact, suggesting the potential for ongoing chronic damage.

Chronic Kidney Disease (CKD) is one of the worldwide health threats, kidneys may be affected by internal pathologies and environmental factors as well as a number of genes are also involved in the kidney dysfunctioning, so UMOD is one of these genes which has significant effect in regulating urea and creatinine level in blood and decreased excretion of uric acid which causes interstitial damages in kidney and may result CKD. Hence, the present study is envisaged to observe the association of this gene locus (r.1264T>C) with CKD in Pakistani population using ARMS-PCR genotyping, for the purpose, a total of 75 samples were investigated, out of which 45 were found homozygous wild-type, 21 heterozygous and 09 homozygous mutants. PLINK data analysis toolset showed that our sampled population does not obeys Hardy-Weinberg Equilibrium by χ² (2, N = 75) = 18.87, p = and Chi-square statistics with p = 1.40×10^-005 showing a significant genotypic association with the subject phenotype with an alternative allele frequency of 0.15 and 0.48 within cases & controls respectively. Furthermore, an odds-ratio of 0.2 predicts that the risk of wild-type allele (A) is 0.8 times higher in patient as compared to controls and mutant allele is 0.2 times preventive in patients. Additionally, a few bioinformatics tools e.g., ProtParam, PsiPred, TMHMM-2.0, Motif Scan, ScanProsite, NetPhos3.1, GPS PAIL2.0, NetOGlyc4.0, PRmePRed, PDB-RCSB and STRING were applied to predict different characteristics of Uromodulin protein including physicochemical properties, secondary/transmembrane structures, protein conserved domains and posttranslational modifications respectively. This pilot scale study provided a comprehensive insight of the association of aforementioned genetic variant with CKD to identify and evaluate the genetic risk factors along with functional effects of the subject variant which may be used as CKD prevention by adopting DNA screening and genetic counselling recommendations.

Background: Pneumocystis pneumonia (PCP) commonly affects immunocompromised individuals, whereas in immunocompetent persons, it occurs relatively rarely, and in most cases the Pneumocystis infection is detected as an asymptomatic colonization. The present study aimed to establish the prevalence of Pneumocystis jirovecii infection in human hosts with different immune status (immunocompromised and immunocompetent), using molecular diagnostic methods, and to compare their diagnostic value with that of classical staining methods. Methods: We used the collected to this moment data from a prospective study on the prevalence of pneumocystosis among the Bulgarian population. Clinical specimens (including throat secretion, induced sputum, tracheal aspirate, and bronchoalveolar lavage collected from 220 patients suspected of PCP (153 immunocompetent and 67 immunocompromised patients) were examined with staining microscopic methods and real-time PCR for detection of P. jirovecii. Results: DNA of the pathogen was detected in 38 (17%) specimens (32 immunocompromised patients and 6 immunocompetent subjects). From all 220 clinical samples examined by staining methods, only in five (2%) P. jirovecii cysts were detected by the Gomori's stain. All patients with PCP were treated with trimethoprim-sulfamethoxazole, but in ten of them (HIV- positive patients) the disease was with fatal outcome. Conclusions: This study is the first for the country including the main available laboratory methods for diagnosis of human pneumocystosis in Bulgaria. Regarding the etiological diagnosis of PCP, in our study the sensitivity of real-time PCR was higher compared to the staining methods. The choice of a method for sample collection and examination has an important role in the efficiency of the laboratory diagnostics.

In Kawasaki disease (KD), convulsions are rare and mostly attributed to latent and coexisting encephalitis/encephalopathy due to KD itself. Therefore, we retrospectively investigated concomitant viral infections in patients with KD. Between January 2012 and December 2020, among 523 patients with KD, 7 (1.53%), 1 (0.19%), 6 and 1 (1.15% and 0.19%) were positive for adenovirus (AdV-Ag), respiratory syncytial virus (RSV-Ag), and influenza A or B (Flu A-Ag or Flu B-Ag) antigens, respectively. Among them, two were positive for both AdV-Ag and FluA-Ag. Seven patients with KD (1.3%) presented febrile convulsions, and only one was diagnosed with encephalitis/encephalopathy. Between January 2021 and August 2022, 57 patients with KD were newly diagnosed among whom FilmArray® respiratory panel (FARP) was applied on 24 patients, and 14 (58.3%) were positive for at least one virus. During the same period, 32 out of 45 patients (71.1%) with febrile convulsions examined using the FARP were positive for at least one virus. The trend of viruses in patients with KD and febrile convulsion reflected the regional epidemic viral infection. Respiratory virus detected at a much higher rate using the FARP suggests that most convulsions in patients with KD might be related to concomitant viral infection. The possible involvement and association of concomitant respiratory viral infections in patients with KD with prognosis or encephalopathy/encephalitis remain to be investigated.

Despite several decades’ effort to detect and identify phytoplasmas (Tenericutes: Mollicutes) using PCR and Sanger sequencing focusing on diseased plants, knowledge of phytoplasma biodiversity and vector associations remains highly incomplete. To improve protocols for documenting phyto-plasma diversity and ecology, we used DNA extracted from phloem-feeding insects and com-pared traditional Sanger sequencing with a next-generation sequencing method, Anchored Hy-brid Enrichment (AHE) for detecting and characterizing phytoplasmas. Among 22 of 180 leaf-hopper samples that initially tested positive for phytoplasmas using qPCR, AHE yielded phyto-plasma 16Sr sequences for 20 (19 complete and 1 partial sequence) while Sanger sequencing yield-ed sequences for 16 (11 complete and 5 partial). AHE yielded phytoplasma sequences for an addi-tional 7 samples (3 complete and 4 partial) that did not meet the qPCR threshold for phytoplasma positivity or yielded non-phytoplasma sequences using Sanger sequencing. This suggests that AHE is more efficient for obtaining phytoplasma sequences. Twenty-three samples with sufficient data were classified into eight 16Sr subgroups (16SrI-B, I-F, I-AO, III-U, V-C, IX-J, XI-C, XXXVII-A), three new subgroups (designated as 16SrVI-L, XV-D, XI-G) and three possible new groups. Our results suggest that screening phloem-feeding insects using qPCR and AHE sequencing may be the most efficient method for discovering new phytoplasmas.

A selection of 36 commercial probiotic fermented dairy products from UK and Europe markets were evaluated for the numbers, types and viability of Lactobacillus strains against the stated information on their packages. A comparative study was carried out on selectivity of MRS-Clindamycin, MRS-Sorbitol and MRS-IM Maltose, to select the right medium for enumeration of probiotic Lactobacillus. Based on selectivity of medium for recovery of the targeted lactobacilli and also simplicity of preparation, MRS-Clindamycin was chosen as the best medium for enumeration of probiotic Lactobacillus in fermented milks. The results of enumeration of lactobacilli showed that 22 out of a total 36 tested products contained more than 106 colony forming units/g at the end of their shelf- life, which comply with the recommended minimum therapeutic level for probiotics. Rep-PCR using primer GTG-5 was applied for initial discrimination of isolated strains, and isolates, which presented different band profile, were placed in different groups. The isolated Lactobacillus spp. were identified mainly as Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus paracasei by analysis of partial sequences of the 16S ribosomal RNA and rpoA genes. In conclusion, it is unknown to recommend the adequate number of probiotic bacteria to be consumed to ensure the beneficial properties.

Although rRT-PCR is the gold standard method for SARS-CoV-2 detection, some factors, such as amplification inhibitors presence, lead to false-negative results. Here we describe differences between rRT-PCR results for SARS-CoV-2 infection in normal and diluted samples, simulating the need for dilution due to amplification inhibitors presence. Viral RNA extraction of nasopharyngeal swabs samples from 20 patients previously detected as 'Negative' and 21 patients detected as 'Positive' for SARS-CoV-2 was realized with the EasyExtract DNA-RNA (Interprise®). rRT-PCR was realized with OneStep/COVID-19 (IBMP) kit with normal and diluted (80µl of H₂O RNAse free) samples, totaling 82 tests. The results indicate that there is an average variation (ɑ < 0.05) delaying Cq between the amplification results of internal control (IC), N Gene (NG), and ORF-1ab (OF) of 1.811 Cq, 3.840 Cq, and 3.842 Cq, respectively. The extraction kit does not completely purify the inhibitor compounds, therefore non-amplification by inhibitors may occur. In this study, we obtained a 19.04% false-negative diagnosis after sample dilution, and this process reduces the efficiency of rRT-PCR to 29.80% for detecting SARS-CoV-2. Knowing the rRT-PCR standards of diluted samples can help in the identification of false-negative cases, and consequently avoid a wrong diagnosis.

An outbreak of winter disease, complicated by severe respiratory syndrome, occurred in January 2020 in a high production dairy cow herd located in a hilly area of the Calabria region. Of the 52 animals belonging to the farm, 5 (9.6%) died with severe respiratory distress, death occurring 3-4 days after the appearance of the respiratory signs (caught and gasping breath). Microbiological analysis revealed absence of pathogenic bacteria whilst Real-time PCR identified the presence of RNA from Bovine Coronavirus (BCoV) in several organs: lungs, small intestine (jejunum), mediastinal lymph nodes, liver and placenta. Since being the only pathogen identified, BCoV was hypothesized to be the cause of the lethal pulmonary infection. Like the other CoVs, BCoV is able to cause different syndromes. Its role in calfhood diarrhoea and in mild respiratory disease is well known: we report instead the involvement of this virus in a severe and fatal respiratory disorder, with symptoms and disease evolution resembling that of Severe Acute Respiratory Syndromes (SARS).

The genus Naegleria, of free-living amoeba (FLA) group, has been investigated mainly due to its human health impact resulting in deadly infections and their worldwide distribution on freshwater systems. Naegleria fowleri, colloquially known as the “brain-eating amoeba”, is the most studied Naegleria species because it causes Primary Amoebic Meningoencephalitis (PAM) of high lethality. The assessment of FLA biodiversity is fundamental to evaluate the presence of pathogenic species and the possibility of human contamination. However, the knowledge of FLA distribution in Brazil is unknown, and to rectify this situation we present a research on identifying Naegleria spp. in the Monjolinho River, as a model study. The river is a public Brazilian freshwater source that crosses the city of São Carlos. Five distinct sampling sites were examined through limnological features, trophozoites culturing and PCR against internal transcribed spacers (ITS) regions and 5.8S rRNA sequence. The results identified N. philippinensis, N. canariensisi, N. australiensis, N.gruberi, N. dobsoni sequences, as well as a Vahlkampfia sequence. The methodology delineated here represents the first Brazilian Naegleria spp. study on a freshwater system. Our result stresses the urgency of a large scale evaluation of the presence of free-living amoebas in Brazil.

Candida species are one of the most important causes of human infections, especially in hospitals and among immunocompromised patients. The correct and rapid etiological identification of yeast infections is important to provide adequate therapy, reduce mortality and control outbreaks. In this study, Candida species were identified in patients with suspected fungal infection, and phenotypic and genotypic identification methods were compared. A total of 167 axenic fungal cultures and 46 clinical samples were analyzed by HardyCHROM®, MicroScan®, and PCR-RFLP. The species of the C. albicans complex were the most frequent, followed by C. tropicalis and C. glabrata. Less common but clinically relevant species of Candida were also isolated. The comparison between the three methods was concordant, especially for the most common Candida species. Fungal DNA amplification was successful in all clinical samples.

The aim of the study was to determine the presence Fusarium species and mycotoxins in winter wheat grain in Poland. Grain samples from different locations in Poland in 2009 and 2010 were analysed for the content of biomass of Fusarium species and mycotoxins. In 2009 biomass of F. graminearum and F. poae was present in all samples, F. culmorum in 82% of samples, F. avenaceum in 55% of samples. F. sporotrichioides, F. tricinctum and F. equiseti were found only in individual samples. F. langsethiae was not detected. In 2010, five Fusarium species were detected with the exception of F. sporotrichioides. The highest content of biomass was found for F. graminearum followed by F. avenaceum, F. poae and F. langsethiae. The amount of F. culmorum biomass was very low. The most frequently occurring species was F. poae and F. graminearum. In 2009, deoxynivalenol was detected in all samples. In 2010, the average content of deoxynivalenol was lower than in 2009. Nivalenol was detected at very low concentration in both years. Significant correlations between content of F. graminearum biomass and deoxynivalenol concentration in grain and between content of F. poae biomass and nivalenol concentration in grain in 2009 were found. The most important finding of this study was that main Fusarium species infecting wheat kernels in Poland in both years was F. graminearum. The amount of biomass of F. graminearum was the highest in both years. It was present in the most samples. The other frequently detected species was F. poae, which in 2010 appeared in more samples than F. graminearum. However, the amount of F. poae biomass was lower. F. culmorum, species that was previously dominating as wheat pathogen in Poland, was found less frequently than F. graminearum. The amount of biomass of this species was the lowest in 2010.

With the rapid progress of genetic engineering and gene therapy, World Anti-Doping Agency has alerted to gene doping and prohibited its use in sports. However, there is no standard method available yet for detection of transgenes delivered by recombinant adenoviral (rAdV) vectors. Here we aimed to develop a detection method for transgenes delivered by rAdV vectors in a mouse model that mimics gene doping. rAdV vectors containing mCherry gene was delivered in mice through intravenous injection or local muscular injection. After five days, stool and whole blood samples were collected, and total DNA was extracted. As additional experiments, whole blood was also collected from mouse tail tip until 15 days from injection of the rAdv vector. Transgene fragments from different DNA samples were analyzed using semi-quantitative PCR (sqPCR), quantitative PCR (qPCR), and droplet digital PCR (ddPCR). In the results, transgene fragments could directly be detected from blood cell fraction-DNA, plasma-cell free DNA and stool-DNA by qPCR and ddPCR, depending on specimen type and injection methods. We observed that a combination of blood cell fraction-DNA and ddPCR was more sensitive than other combinations used in this model. These results could accelerate the development of detection methods for gene doping.

Numerous protocols have been published for extracting DNA from phlebotomines. Nevertheless, their small size is generally an issue in terms of yield, efficiency, and purity, for large-scale individual sand fly DNA extractions when using traditional methods. Even though this can be circumvented with commercial kits, these are generally cost-prohibitive for developing countries. We encountered these limitations when analysing parasite infection in Lutzomyia spp. by PCR [1] and, for this reason, we evaluated various modifications on a previously published protocol ([2] and Acardi personal communication). The most significant variation was the use of a different lysis buffer [3] to which added Ca2+ (buffer TESCa), because this ion protects proteinase K against autolysis, increases its thermal stability, and could have a regulatory function for its substrate-binding site [4]. Individual sand fly DNA extraction success was confirmed by amplification reactions using internal control primers that amplify a fragment of the cacophony gene [5,6]. To the best of our knowledge, this is the first time a lysis buffer containing Ca2+ has been reported for the extraction of DNA from sand flies.

We present a case of Capnocytophaga canimorsus septic shock after a dog bite in an immunocompetent individual, where real-time nanopore metagenomic sequencing characterized the microbial agent within 19 hours, with subsequent confirmation using droplet digital PCR. Oral swabs from the dog demonstrated a nearly-identical C. canimorsus isolate by sequencing.

Acid lime [Citrus aurantifolia (Christm.) Swingle] is a fruit crop, enriched with high commercial value and is cultivated in 60 out of 75 districts representing all geographical landscapes of Nepal. Lack of high yielding cultivars is probably one of the main reason for its extremely reduced productivity which warrants a deep understanding of genetic diversity in existing germplasm. Hereby, we aim to access the genetic diversity of acid lime germplasm cultivated at 3-different ecological gradients of eastern Nepal employing PCR-based Inter-Simple Sequence Repeats markers (ISSR). Altogether, 21 polymorphic ISSR markers were used to assess the genetic diversity in 60 acid lime cultivars sampled from different geographical locations. Analysis of binary data matrix was performed on the basis of bands obtained, scoring of the data was done accordingly, and principal coordinate analysis and phenogram were constructed using different computer algorithms. ISSR profiling yielded 234 amplicons, of which 87.18% were found to be polymorphic. The number of amplified fragments ranged from 7-18 with amplicon size ranging from 250-3200 bp. The NTSYS based Cluster analysis using UPGMA algorithm taking Dice Similarity coefficient separated 60 accessions into 2-major and 3-minor clusters. The genetic diversity analysis revealed the highest for Terai and the lowest for High-hill zone. Cluster I comprised of accessions from High-hill and Mid-hill regions revealing the close genetic relationship, whereas cluster II comprised of accessions from all three agro-ecological zones and the exotic varieties. Furthermore, our results revealed the accessions harvested from different geographical gradients were not genetically distinct, but highest diversity was observed in Terai accessions in comparison to the regions belonging to the High and Mid-hills. Thus, our data indicate that the ISSR provides a better option for evaluating the genetic diversity of Nepalese Acid Lime cultivars and furnished significant information, assisting parental selection in current and future breeding programs and germplasm conservation which ultimately may help to provide a technological breakthrough for the farmers of the developing country like Nepal.

Autonomic dysregulation is associated with cardiovascular consequences in obstructive sleep apnea (OSA). This study aimed to investigate the effect of acute continuous positive airway pressure (CPAP) treatment on autonomic activity and to identify factors contributing to the heart rate variability (HRV) changes in OSA. Frequency domain HRV parameters were calculated and compared between the baseline polysomnography and during the CPAP titration in 402 patients with moderate to severe OSA. There were significant reductions in total power, very low-frequency band power, low-frequency band power, and high-frequency band power during the CPAP titration as compared to the baseline polysomnography. This tendency was more pronounced in men than in women, and in patients with severe OSA than those with moderate OSA. Multivariate analysis found that changes in apnea-hypopnea index and oxygen saturation were significantly associated with changes in sympathetic and parasympathetic activity, respectively. This study demonstrated that HRV parameters significantly changed during the CPAP titration, indicating a beneficial effect of CPAP in restoration of sympathetic and parasympathetic hyperactivity in OSA. Prospective longitudinal studies should determine whether long-term CPAP treatment aids in maintaining the long-lasting improvement of the autonomic functions, thereby contributing to the prevention of cardiovascular and cerebrovascular diseases in patients with OSA.

In this study, we investigate the existence of positive solutions within a system of Riemann-Liouville fractional differential equations that incorporate the (r1,r2,r3)-Laplacian operator while being subject to three-point boundary conditions. These equations incorporate various fractional derivatives and are influenced by parameters represented as (ψ1,ψ2,ψ3). Our approach involves employing techniques such as cone expansion and compression of the functional type, in conjunction with the Leggett-Williams fixed point theorem, to establish the existence of positive solutions. To emphasize the practical significance of our findings in the realm of fractional differential equations, we provide two illustrative examples.

Background: To examine the effects of continuous positive airway pressure (CPAP) therapy on central corneal thickness (CCT) in patients with obstructive sleep apnea (OSA) followed for 12 months. Methods: Participants were patients with OSA and an indication for CPAP who were enrolled after diagnosis and graded according to the apnea hypopnea index (AHI) determined in an overnight polysomnography. Each subject underwent a full ophthalmologic examination including ultrasound pachymetry (USP) with an SP3000 USP (Tomey Inc) at the time points just before CPAP onset, and after 3 and 12 months of CPAP treatment. Results: The study sample comprised 40 eyes of 20 patients. Adherence to CPAP was adequate (5.23 ± 1.77 h/night). After receiving 12 months of CPAP, positive correlation was observed between hours of CPAP use and CCT (r = 0.382, p = 0.015). Intermittent hypoxemia pre-CPAP or hypoxia severity pre-CPAP were also positively correlated with the CCT recorded after 12 months of CPAP (r = 0.394, p = 0.012; r = 0.324, p = 0.041). Significant correlations were, however, not observed after only 3 months of CPAP treatment. In addition, compared to measurements at 3 months, intraocular pressure was higher after 12 months of treatment. Conclusions: Our findings indicate that 12 months of correct adherence to CPAP are needed to detect significant CCT thickening, and that the greater the pre-CPAP hypoxemia, the greater the capacity for an increase in CCT to be produced in response to 12 months of this therapy.

Quartz Crystal Microbalance (QCM) is a well-known method to measure the mass of ligands bound to receptor-covered surfaces in various fields, ranging from materials science to biology. Actin molecule is essential in various cellular processes through its function of monomer-to-filament polymerization, forming a double-stranded filament with a 75 nm pitch. In this study, QCM was applied to measure the physical properties of actin, finding that the frequency shifted negatively when monomeric actin bound to the QCM surface, whereas the frequency shifted positively when filaments bound. Using fluorescence light microscopic observations, we have examined whether negative and positive frequency shifts originated from their length. In order to control this process, a severing protein, fragmin was used to change the length of actin filament. When the average length of filaments shortened, the magnitude of positive shift decreased in a concentration-dependent manner. However, shorter filaments showed a negative frequency shift depending on their concentration, showing that the weight derived from the loaded concentration and the function derived from actin polymerization dynamics are detectable. Interestingly, the transition from positive to negative occurred when the average length of actin was ~33 nm, or approximately equal to the half-pitch of filaments. Thus, actin filaments can serve as an excellent standard to measure the mechanical properties of biopolymers. Our results show that the QCM sensor could be both a "weight" and a "function" sensor of biomolecules in vitro.

This paper presents the goals and components of a quantitative energy balance assessment framework to define PEDs flexibly in three important contexts: the context of the district's density and RES potential, the context of a district's location, induced mobility and the context of the dis-trict's future environment and its decarbonized energy demand or supply. It starts by introducing the practical goals of this definition approach: achievable, yet sufficiently ambitious to be inline with Paris 2050 for most urban and rural Austrian district typologies. It goes on to identify the main design parts of the definition: system boundaries, balancing weights and balance targets and argue how they can be linked to the definition goals in detail. In particular we specify three levels of system boundaries and argue their individual necessity: operation, including everyday mobili-ty, including embodied energy and emissions. It argues that all three pillars of PEDs, energy effi-ciency, onsite renewables and energy flexibility can be assessed with the single metric of a prima-ry energy balance when using carefully designed, time-dependent conversion factors. Finally, it is discussed how balance targets can be interpreted as information and requirements from the sur-rounding energy system, which we identify as a "context factor". Three examples of such context factors, each corresponding to the balance target of one of the previously defined system bounda-ries operation, mobility and embodied emissions are presented: Density (as a context of opera-tion), sectoral energy balances and location (as a context for mobility) and an outlook of a person-al emission budgets (as a context for embodied emissions). Finally, the proposed definition framework is applied to seven distinct district typologies in Austria and discussed in terms of its design goals.

Background: Histoplasmosis is a systemic mycosis, present globally. We aimed to describe cases of histoplasmosis (Hc) and to establish a risk profile associated to Hc in HIV-infected patients (HIV+). Methods: This was a retrospective study of patients with a clinical-. laboratorial diagnosis of Hc. Data was fed into REDCap and statistical analysis was done with R . Results: We included 99 records, 65 HIV+ and 34 HIV-. Average age was 39 years. Median time from onset to diagnosis was 8 weeks in HIV- and 22 weeks in HIV+. Disseminated histoplasmosis occurred in 79.4% HIV+, vs 36.4% HIV-. Median CD4 count was 70. Co-infection with tuberculosis was present in 20% of HIV+. Blood cultures were positive in 32.3% of HIV+ vs 11.8% of HIV- (p=0.025); bone marrow culture was positive in 36.9% vs 8.8%(p=0.003). Most HIV+ patients (71.4%) were hospitalized. On univariate analysis, anemia, leukopenia, intensive care, use of vasopressors and mechanical ventilation were associated with death in HIV+. Conclusions: Most of our patients with histoplasmosis were HIV+, presenting advanced AIDS. Diagnosis was late in HIV+ patients, and they frequently presented disseminated Hc, required hospitalization, and died. Early screening for Hc in HIV+ and drug induced immunosuppressed patients is crucial.

Positive invariant set is an important concept of dynamic systems. The purpose of this paper is to study the sufficient and necessary conditions that the set of ellipsoids and the Lorenz cone are positive invariant sets of discrete time systems. By means of nonlinear programming and induced norm, the problem of positive invariance is formulated as an optimization problem, and the equivalent dual form optimization is also presented using the dual optimization method. Our results provide more alternative methods for determining the positive invariance of quadratic form convex sets from the point of view of optimization and dual optimization. The effectiveness of this method is demonstrated by numerical examples.

ABSTRACT: Psychological vulnerability (PV) and low mental health literacy correlate negatively with higher education students' positive mental health and health behaviours. Levels of mental health literacy limit self-help, adaptative coping strategies and academic success. However, few studies examine the significant differences between students' PV, mental health literacy, positive mental health (PMH) and modifiable behaviours variables. Identifying modifiable vulnerability factors is essential when seeking and providing professional help. Method: A cross-sectional descriptive study was conducted to investigate the differences between modifiable behaviours and PV, mental health literacy, and PMH in higher education students. A sample of 3,600 undergraduate students was recruited, mostly female (78.8%), with an average age of 23 years (SD = 6.68). Results: The majority were women, did not exercise, were dissatisfied with sleep quality time per night (62.1% sleep less than 7 hours per night) and did not engage in leisure activities. Also, most respondents had PV and low literacy levels, particularly at F1, and 67.9% scored in the flourishing group. Results showed significant differences (p<0.001) between gender, age, sleep, exercise, diet, leisure activities and PV, and PMH and literacy. Conclusions: The results identified modifiable vulnerability factors that underpin the design of intervention-targeted programmes to promote literacy and PMH in higher education campuses in Portugal to foster students’ well-being and self-help strategies.

Aboriginal Australians have a fundamental human right to opportunities that lead to healthy and flourishing lives. While the impact of trauma on Aboriginal Australians is well-documented, a pervasive deficit narrative that focuses on problems and pathology persists in research and policy discourse. This narrative risks further exacerbating Aboriginal disadvantage, through a focus on ‘fixing what is wrong’ with Aboriginal Australians, and the internalising of these narratives by Aboriginal Australians. While a growing body of research adopts strength-based models, limited research has sought to explore Aboriginal flourishing. This conceptual paper seeks to contribute to a burgeoning paradigm shift in Aboriginal research, seeking to understand what can be learned from Aboriginal people who flourish, how we best determine this, and in what contexts this can be impactful. Within, we argue the case for a new approach to exploring Aboriginal wellbeing that integrates salutogenic, positive psychology concepts with complex systems theory to understand and promote Aboriginal wellbeing and flourishing, While deeper work may be required to establish the parameters of a strength-based, culturally aligned Aboriginal conceptualisation of positive psychology, we suggest the integration of Aboriginal and Western methodologies offer unique and potent means of shifting the dial on seemingly intractable problems.

Positive education is seen as a transformative methodological approach capable of improving the act of teaching and learning and, above all, essential for the development of students' personal skills and competences. However, few studies have been carried out on this subject in the field of lifelong and adult education. This study works with a sample of 399 people over 16 years of age and students of the Universidad Popular de Dos Hermanas in order to show the relationship between the Pygmalion effect and learned helplessness in the process of acquiring knowledge in adulthood. In this way, three tools were used: a questionnaire that showed teachers' perceptions of students' qualities and behaviour and two that provided information on self-concept, self-esteem, personal and social skills and other variables directly related to emotional intelligence and positive education. It shows how exposure to negative operational conditioning factors influences the psychosocial and socio-educational development of students in every possible way, while on the other hand, it indicates the importance of positive education to compensate for this phenomenology by improving the development and growth of those who study and participate in non-formal education. Likewise, the factorial interrelation of both positive and negative conditioning factors and their incidence on learning is shown; the importance of neutralising the negative components and strengthening the positive reinforcement and the role played by the community and education professionals as catalysts and behavioural modulators at any stage of learning and age group for the achievement of the objectives of the student and of education itself in a broad sense

Since SARS-CoV-2 has spread all over the world, one of the key questions is how long immunity works for protecting hosts from infection. However, it was difficult to distinguish reinfection from re-detected positive cases caused by other reasons. We confirmed 23 re-detected positive cases in Sapporo City in 2020, and among them, the case with 205-days interval from initial onset to the second confirmation date was found. We calculated the probability of observing more than one case with over d-days interval of re-detection, pobs, to find out whether the case with 205-days interval had the similar epidemiological characteristic with others or not. We found that 5th percentile and its 95% confidence interval of pobs was 118 (45, 216) for Sapporo City data and 92 (86, 98) for KCDC data. This result indirectly suggested the case with 205-days interval was reinfection.

Autophagy is an evolutionarily conserved central process in host metabolism. Among its major functions are conservation of energy during starvation, recycling organelles, and turnover of long-lived proteins. Besides, autophagy plays a critical role in removing intracellular pathogens and very likely represents a primordial intrinsic cellular defence mechanism. More recent findings indicate that it has not only retained its ability to degrade intracellular pathogens, but also functions to augment and fine tune antiviral immune responses. Interestingly, viruses have also co-evolved strategies to manipulate this pathway and use it to their advantage. Particularly intriguing is infection-dependent activation of autophagy with positive stranded (+)RNA virus infections, which benefit from the pathway without succumbing to lysosomal degradation. In this review we summarise recent data on viral manipulation of autophagy, with a particular emphasis on +RNA viruses and highlight key unanswered questions in the field that we believe merit further attention.

Building on the Sustainable Happiness Model, this study examined how congruency between ideal self-image and brand image influence a sense of happiness. The findings show that when ideal self-image and ideal social self-image are congruent with brand image a sense of happiness can be enhanced through brand identification and positive emotions. This study contributes to literature as it reveals the mechanism of how congruency between ideal self-image and brand image positively affect happiness.

The global craft beer market is witnessing substantial growth, with a particular spotlight on sour beers. These unique brews are created through lactic fermentation initiated by lactic acid bacteria (LAB). However, the presence of hops, renowned for their antimicrobial properties, can pose a limitation within the realm of sour beer production. This research aims to explore and assess the potential of sour beer as a probiotic beverage. It involves a comparative analysis of the viability of Lacticaseibacillus paracasei subsp. paracasei F19 (F19) and 431 (L431) in a sour beer with an International Bitterness Unit (IBU) value of 26.65, alongside examining the ex-pression of genes such as horA, horB, horC, hitA, bsrA, and recA, which are associated with hop resistance. The study encompasses the development of two formulations of sour beers characterized by both high hop content and a robust probiotic count (8.44 – 8.77 log CFU/mL). The findings suggest that both probiotic strains of F19 and L431, are well-suited to produce sour beers with elevated hop levels, demonstrating excellent stability. Notably, the expression of genes responsible for hops resistance exhibited distinct modulation patterns among the two strains. It appears that a higher concentration of bsrA, as observed in strain L431, may be more effective in mitigating the effects of hop-related stress, potentially surpassing the combined expression of horA and bsrA. Nonetheless, further research is essential to validate this observation in gene expression. F19 and L431 emerge as promising candidates for the development of sour beers with increased hop intensity.

The COVID-19 pandemic caused by the SARS-CoV-2 virus has inflicted significant mortality and morbidity worldwide. Continuous virus mutations have led to the emergence of new variants. The Omicron BA.1 sub-lineage prevailed as the dominant variant globally at the beginning of 2022 but was subsequently replaced by BA.2 in numerous countries. Wastewater-based epidemiology (WBE) offers an efficient tool for capturing viral shedding from infected individuals, enabling early detection of potential pandemic outbreaks without relying solely on community cooperation and clinical testing resources. This study integrated RT-qPCR assays for detecting general SARS-CoV-2 and its variants levels in wastewater into a modified triple susceptible-infected-recovered-susceptible (SIRS) model. The emergence of the Omicron-BA.1 variant was observed, replacing the presence of its predecessor, the Delta variant. Comparative analysis between the wastewater data and the modified SIRS model effectively described the BA.1 and subsequent BA.2 waves, with the decline of the Delta variant aligning with its diminished presence below the detection threshold in wastewater. This study demonstrates the potential of WBE as a valuable tool for future pandemics. Furthermore, by analyzing the sensitivity of different variants to model parameters, we are able to deduce real-life values of cross-variant immunity probabilities, emphasizing the asymmetry in their strength.

Meloidogyne javanica causing root-knot nematode in soybean is an important problem in soybean areas, leading to several yield losses. Some accessions have been identified carrying resistance loci to this nematode specie. In this study, a set of 317 soybean accessions were characterized for resistance to M. javanica. Genome-wide association study (GWAS) was performed using SNPs from genotyping-by-sequencing (GBS), and a region of 29.2 Kbp on chromosome 13 was identified. The haplotype analysis showed that SNPs were able to discriminate susceptible and resistant accessions, leading to 25 accessions sharing the resistance locus. Furthermore, 5 accessions may be new M. javanica resistance sources. The screening of the SNPs in the USDA soybean germplasm showed that several accessions previous reported as resistance to other nematodes also showed the resistance haplotype on chromosome 13. High levels of concordance among the phenotypes of Brazilian cultivars and the SNPs in chromosome 13 were observed. A in silico analysis of the mapped region on soybean genome revealed a presence of 5 genes with structural similarity with major resistance genes. The expression levels of the candidate genes in the interval demonstrated a potential pseudogene, and other two model genes up-regulated in the resistance source after pathogen infection. The SNPs associated to the region conferring resistance is a important tool for introgression of the resistance by marker-assisted selection in soybean breeding programs.

Secondary metabolites are bioactive compounds produced by living organisms that can reveal symbiotic relationships in nature. In the work described here, soil-born entomopathogenic-nematodes associated with symbiotic bacteria (Xenorhabdus stockiae and Photorhabdus luminescens) extracted from solvent supernatant containing secondary metabolites showed significant inhibitory action against Escherichia coli, Staphylococcus aureus, Enterococcus faceium, Proteus vulgaris, Bacillus cereus, Bacillus subtilis. These secondary metabolites, characterized by Fourier transform infrared spectroscopy were found to be amine groups of proteins; hydroxyl and carboxyl groups of polyphenols; hydroxyl groups of polysaccharides, and carboxyl groups of organic acids. The major compounds identified by Gas chromatography-mass spectrometry analysis of ethyl acetate extracts of Xenorhabdus stockiae revealed the existence of Nonanoic acid derivatives, Paromycin, Pyrrolidinone, Octodecanal derivatives, Trioxa-5-aza-1-silabicyclo, 4-Octadecenal, Methyl ester, Oleic Acid and 1,2-benzenedicarboxylicacid. Additional extraction from Photorhabdus luminescens was expressed functional compounds such as Indole-3-acetic acid, Piperidinol derivatives, Phthalic acid, 1-Tetradecanol, Nemorosonol, 1-eicosanol and unsaturated fatty acids. These outcomes support the development of new alternative natural antimicrobial agents for pathogen suppression in the future.

Several pathotypes of enteric E. coli have been identified. The group represented by Shiga toxin-producing E. coli (STEC) is of particular interest. Raw milk and raw milk products are significant sources of STEC infection in humans; therefore, identifying pathogens at the herd level is crucial for public health. Most national surveillance programs focus solely on raw milk and raw milk cheeses that are ready for retail sale, neglecting the possibility of evaluating the source of contamination directly at the beginning of the dairy chain. In order to assess the viability of new molecular methodologies applied to raw milk filters enabling the identification of the presence of STEC serotypes in milk production and apply the same methods to identify the presence of these pathogens in calf feces we analyzed, through a commercial real-time PCR assay, a total of 290 samples coming from 18 different dairy herds; 88 bulk tank milk (BTM), 104 raw milk filters (RMF) and 98 calves’ feces samples. In total 3.4% of BTM, 41.4% of RMF and 73.4% of calves’ feces tested positive for stx presence, supporting our hypothesis that BTM is not a suitable matrix to assess the STEC presence at herd level, underestimating it. Our conclusion is that the surveillance program needs critical and extensive improvements such as RMF and calves’ feces analysis implementation in order to be more efficient in detecting and preventing STEC infections. The epidemiology of these infections and the characteristics of the pathogen clearly show how a One Health approach will be pivotal to improve our capabilities to control the spread of these infections.

Lumpy skin disease (LSD) is a viral disease of cattle and water buffalo characterized by cutaneous nodules, biphasic fever and lymphadenitis. LSD is endemic in Africa and Middle East but in recent years has spread in different Asian countries. Drop of animal productions, vaccination campaign, surveillance costs and trade restrictions have significant impact on the economy of affected countries. The disease is well characterized in cattle while little is known in buffaloes and no experimental studies have been conducted in this species. Six buffaloes and two cattle were inoculated with an Albanian LSD virus (LSDV) field strain. Over a period of 42 days animals were clinically monitored and sampled for serological and virological analysis. Only two buffaloes showed clinical sign, fever, skin nodules and lymphadenitis. All samples tested in real time PCR (blood, swabs, biopsies and organ) gave negative results. Three buffaloes seroconverted at day 39 and 42 post-inoculation, detected by ELISA, but all sera were negative in serum neutralization test. Cattle showed more severe clinical signs. Moreover, viraemia and virus shedding were proven by positive PCR results and antibodies were detected by both ELISA and serum neutralization test. These findings demonstrated the limited susceptibility of buffaloes compared to cattle once experimental infected with LSDV. Further studies are needed to investigate the epidemiological role of the buffaloes.

The early detection of cognitive decline and timely non-pharmacological management or drug therapy are extremely important in providing care for Parkinson’s disease with dementia (PDD). In this study, we first conducted a discovery study to identify six plasma microRNAs that may allow for the differentiation of PD with or without mild cognitive impairment via NGS. A total of 120 participants were further recruited in a validation cohort and divided into four subgroups, namely, normal controls (HC), PD with no dementia (PDND), PD with mild cognitive impairment (PD-MCI) and PDD. Among the six candidates, miR-203a-3p was successfully detected in the plasma of the validation cohort using droplet digital PCR (ddPCR). Our results show that the ratio of miR-203a-3p/miR-16-5p observed in PDD was significantly increased compared to in PD-MCI (p < 0.001) and PDND (p = 0.041). Moreover, the ratio of miR-203a-3p/miR-16-5p showed a significant correlation with MoCA scores (r = -0.237, p = 0.024) in patients with PD (PwP). The ROC curve of the logistic regression model, consisting of the variables of age, the ratio of miR-203a-3p/miR-16-5p and the UPDRS III score, also demonstrated an average AUC of 0.883 via 5-fold cross-validation. In conclusion, the ratio of miR-203a-3p/miR-16-5p may serve as a potential biomarker for distinguishing cognitive dysfunction from PwP.

Campylobacter is one of the most commonly reported foodborne bacteria worldwide. Although C. jejuni and C. coli have been reported to be responsible for the great majority of campylobacterio-sis, the burden of infections by species other than C. jejuni and C. coli have been increasing as a result of a transition to diagnostic test methods that enable the isolation of emerging species. The aim of the present study was to recover C. jejuni, C. coli, and emerging species from the stool samples of 500 patients with gastroenteritis and 100 healthy subjects by use of a filtration meth-od and culture techniques using Butzler agar and mCCDA under microaerophilic or hydrogen enriched atmosphere, identify the species by multiplex PCR methods and assess the significance of emerging species in enteric diseases. Thirty-one (6.2%) Campylobacter spp. were isolated from the stool samples of diarrheic patients but none from healthy individuals. Of 31 isolates, 21 (67.8%), 9 (29%) and 1 (3.2%) were identified as C. jejuni, C. coli and C. concisus by multiplex PCR, respectively. Filtration method was superior to the culture technique using mCCDA under mi-croaerophilic atmosphere. C. concisus was evaluated as the etiology of gastroenteritis as a result of laboratory and clinical evaluations. The present study was the first to indicate that emerging Campylobacter species are rarely detected and C. concisus is linked to acute gastroenteritis in Tur-key where additional studies are warranted to clarify the significance of emerging species in gastroenteritis.

Ostreid herpesvirus 1 (OsHV-1) is one of the most economically important pathogens of Pacific oysters. Understanding the pathogenesis of this virus is critical to developing tools to control outbreaks on shellfish farms. OsHV-1 is genetically related to vertebrate herpesviruses, which have a lytic and a latent stage, with the latent stage capable of being reactivated to the lytic stage. Here, OsHV-1 latency in Pacific oysters was investigated in experimentally and naturally infected oysters. Lytic infection in one-year-old oysters injected with the Tomales Bay strain of OsHV-1 was detectable between 1 and 4 days post-infection (dpi) but was not detectable after 5 dpi. The infected oysters shed 1102 to 1104 DNA copies/ml during the 4-day acute phase. At 21 dpi, the recovered oysters were temperature and chemically stressed and were found to shed 1104 to 1105 DNA copies/ml over a period of 48 h. Lytic shedding was not detectable in two-year-old oysters injected similarly with the same strain of OsHV-1; however, the OsHV-1 genome was detectable by qPCR in the adductor muscle, gill, mantle, and hemocytes within the first 3 dpi, after which it became undetectable. No OsHV-1 was detectable in the adductor muscle, gill, or mantle from experimentally infected oysters on days 15 and 21 post-infection or from oysters sampled 9 months after surviving an OsHV-1 mortality event; however, OsHV-1 DNA could be detected in hemocytes of both experimentally infected oysters at 21 dpi and naturally infected oysters. In addition, lytic viral gene transcription was detectable in hemocytes of experimentally infected oysters between 1 and 21 dpi and in hemocytes of naturally infected oysters.

Morphological features and composition of ISSR DNA fragments were studied in species of the genus Tulipa L. A comparative analysis of the data obtained was carried out and an attempt was made to clarify the species belonging of plants. Plants obtained from various sources were studied. It was found that representatives of the genus Tulipa L. from the two subgenera Tulipa and Eriostemones are well separated not only morphologically, but also by the composition of ISSR fragments. At the intra- and interspecific level, the results for morphological traits and molecular data differed. Interspecific and intraspecific differences were more clearly traced in the complex analysis of morphological features and ISSR PCR data. All samples obtained in the form of bulbs and renewed vegetatively had an identical set of ISSR fragments. Plants grown from seeds were characterized by a significant variety of molecular markers and had both species-specific and in-dividual genetic variability. It was found that the samples obtained and registered as the Tulipa urumiensis Stapf are the yellow-flowered form of the Tulipa tarda Stapf, and a sample received as the Tulipa turkestanika Regel is the Tulipa bifloriformis Vved. Thus, a comprehensive study of both the data of fragmentary DNA analysis and morphological features, provided a sufficient number of DNA samples, makes it possible to clarify the species of tulip plants, and also allows us to assess the genetic diversity of the genus Tulipa.

1) Background: Northwest Syria (NWS), is a conflict-affected and unstable area. Due to its limited health infrastructure, accessing advanced COVID-19 testing services is challenging. COVID-19 antigen rapid diagnostic tests (Ag-RDTs) have the potential to overcome this barrier. Therefore, a pilot project was implemented to introduce Ag-RDTs in this setting aiming to a) describe the feasibility, uptake, and results of Ag-RDTs; and b) identify facilitators and barriers to Ag-RDTs testing. 2) Methods: A cross-sectional study design involving secondary analysis of data collected during the project’s monitoring was developed. A local NGO implemented 25,000 Ag-RDTs cross boarder through trained community health workers. 3) Results: A total of 27,888 persons were found eligible and enrolled, of which 24,956 (89.5%) consented to test and 121 (0.5%) were found positive. Highest positivity was observed among those with severe COVID-19 symptoms (12.7%), those with respiratory illnesses (2.5%), persons enrolled at Afrin Hospitals (2.5%), and healthcare workers (1.9%). A non-random sample of 236 people underwent confirmatory rt-PCR test. Accordingly, observed sensitivity, specificity, positive and negative predictive values, were 80.0%, 96.1%, 91.4% and 90.3%, respectively. Key encountered challenges included obtaining informed consent and conducting confirmatory rt-PCR testing. 4) Conclusion: This project demonstrated great feasibility utilizing Ag-RDTs as a screening/diagnostic tool for COVID-19 infections with nearly 90% uptake. Considering the high specificity and negative predictive values and the higher positivity rates among severe COVID-19 symptomatic, embedding Ag-RDTs into COVID-19 testing strategies for ruling out and in COVID-19 infections would hold a great advantage.

Enteroaggregative Escherichia coli (EAEC) are highly heterogeneous in virulence; we wanted to understand the pathogenic potential of EAEC isolated various clinical and non-clinical sources in an animal model. We infected male BALB/c mice in six mice/group with 50 EAEC isolates, isolated from clinical and non-clinical sources. We studied colonization, weight loss, stool shedding, inflammatory markers and its relationship with 21 virulence genes and phylogroups, EAEC organ burden and histopathological changes. We detected significantly more inflammatory changes and fecal lactoferrin and calprotectin levels in mice infected with EAEC isolated from symptomatic cases. In clinical EAEC isolates, the presence of chromosomal genes {(aap (46%), aaiC (23.3%), and SPATEs (pet (13.3%), sat (20%), sigA and pic (6.6%)}, adhesive variants {(agg4A (53.3%), aggA (53.3%), aafA (36.6%), agg3A (40%)} of EAEC, and master regulator gene aggR (66.6%) were associated with higher levels of lactoferrin and calprotectin. Also, 70% (9/13) of EAEC isolated from acute diarrheal cases bearing chuA (70%) in our study were assigned to group B2 (4 isolates) and D (5 isolates). Real-time PCR analysis revealed that colonization by EAEC strains from different clinical and non-clinical sources occurs up to 10-15 days of life. Even from non-diarrhoeal stools and non-clinical sources, had the potential to cause prolonged colonization, weight loss, and inflammation in the intestine though the degree varied. Moreover, better understanding of EAEC pathogenic pathways is desperately needed in different clinical scenarios.

RT-PCR tests have become the gold standard for detecting the SARS-CoV-2 virus in the context of the COVID-19 pandemic. Because of the extreme number of cases in periodic waves of infection, there is a severe financial and logistical strain on diagnostic laboratories. For this reason, alternative implementations, and validations of academic protocols, that employ the lowest cost and most widely available equipment and reagents found in different regions, is essential. In this study, we report an alternative implementation of the EUA 2019-nCoV CDC assay which uses a previously characterized duplex PCR reaction for the N1 and RNAse P target regions and an additional uniplex reaction for the N2 target region. Taking advantage of the Abbott m2000 Sample Preparation System and NEB Luna Universal Probe One-Step RT-qPCR kit, some of the most widely available and lowest cost nucleic acid extraction and amplification platforms, this modified test shows a state-of-the-art analytical and clinical sensitivities and specificities, when compared with the Seegene Allplex-SARS-CoV-2 assay. This implementation has the potential to be verified and implemented by diagnostic laboratories around the world to guarantee low-cost RT-PCR tests that can take advantage of widely available equipment and reagents.

Peanut (Arachis hypogaea) contains allergenic proteins, which make it harmful to the sensitised population. The presence of peanut in foods must be indicated on label, to prevent accidental consumption by allergic population. In this work, we use chloroplast markers for specifically detection of peanut by real-time PCR, in order to increase the assay sensitivity. Binary mixtures of raw and processed peanut flour in wheat were performed at concentrations ranging from 100000 to 0.1 mg/kg. DNA isolation from peanut, mixtures and other legumes was carried out following three protocols for obtaining genomic and chloroplast-enrich DNA. Quantity and quality of DNA was evaluated, obtaining better results for protocol 2. Specificity and sensitivity of the method has been assayed with specific primers for three chloroplast markers (mat k, rpl16 and trnH-psbA) and Ara h 6 peanut allergen-coding region was selected as nuclear low-copy target and TaqMan probes. Efficiency and linear correlation of calibration curves were within the adequate ranges. Mat k chloroplast marker yielded the most sensitive and efficient detection for peanut. Moreover, detection of mat K in binary mixtures of processed samples was possible up to 10 mg/kg even after boiling, and autoclave 121°C 15 min, with acceptable efficiency and linear correlation. Applicability of the method has been assayed in several commercial food products.

Early identification of fetal abnormalities is a huge challenge for modern obstetrics. Quantitative fluorescent polymerase chain reaction (QF-PCR) has quickly become an effective means of chromosome anomaly detection due to its advantages in terms of timing, manpower and accuracy. The QF-PCR results also make a significant change in clinicians’ attitude to some extent, assisting them providing parents with professional and valuable advice on pregnancy management. In this review, the advantages and drawbacks of QF-PCR will be explored. By reviewing studies published in Vietnamese Medical Journals, we conclude QF-PCR can become a potential screening test in the field of prenatal diagnosis.

Droplet digital polymerase chain reaction (ddPCR) is a method used to detect and quantify nu-cleic acids even when present in exceptionally low numbers. While it has proven to be valuable for clinical studies, it has failed to be widely adopted for environmental and applied studies. Due to the complexity of the chemical and biological composition of environmental samples, protocols tailored to clinical studies are not appropriate, and results are difficult to interpret. We used en-vironmental DNA samples originating from field studies to determine a protocol for environ-mental samples. Samples included field soils which had been inoculated with the soil fungus Rhizophagus irregularis (environmental positive control), field soils that had not been inoculat-ed and the targeted fungus was not naturally present (environmental negative control), and root samples from both field categories. To control for the effect of soil inhibitors, we also in-cluded DNA samples of an organismal control extracted from pure fungal spores (organismal positive control). Finally, we included a no-template control consisting only of the PCR reaction reagents and nuclease free water instead of template DNA. Using original data, we examined which factors contribute to poor resolution in root and soil samples and propose best practises to ensure accuracy and repeatability. Furthermore, we evaluated manual and automatic threshold determination methods and we propose a novel protocol based on multiple controls that is more appropriate for environmental samples.

Streptococcal infection is a main infectious diseases for farmed grey mullet (Mugil cephalus). This study were to identify spreptococcal species in diseased farmed grey mullet and to investigate differences in susceptibility to 13 antibiotics and in genotypes between the stains from the grey mullet and non-grey mullet. 170 samples from diseased farmed grey mullet were collected from three county in 2013 -2016. Multiplex PCR identified L. garviea (146) as the main pathogen, S. agalactia (9), S. dysgalactiae (19), and double infection (5), but no S. iniae. The prevalence changed annually and differed among three counties. Pulsed-field gel electrophoresis (PFGE) analysis demonstrated identical genotype with an ApaI-digested DNA pattern. Disc diffusion results demonstrated differences in antibiotic susceptibility between the strains from grey mullet (146) and non-grey mullet (30). Almost all strains resisted to clindamycin and all strains were susceptible to six antibiotic in grey mullet and 4 antibiotics in non-grey mullet. The reduced susceptible strains was more in non-grey mullet than grey mullet group. The reduced susceptible strains were observed the highest in 2014 and in Chiayi county and decreased from 2014 to 2016. However, the strains with reduced susceptibility to ceftriaxone, cirpofoxacin, moxifloxacin, tetracycline for human treatment were observed.

Toxoplasmosis is a neglected anthropozoonosis caused by the obligate intracellular protozoan, Toxoplasma gondii. The role of non-human primates in the epidemiology of human toxoplasmosis is not clear. Acute and highly fatal cases of toxoplasmosis are frequently reported in new world monkeys and asymptomatic infections in old world monkeys. Here we report detection of latent natural T. gondii infection in olive baboons during a screening exercise to select animals for an experimental toxoplasmosis study. Archived serum samples from 32 olive baboons (Papio anubis), 23 wild caught and nine colony-born, were screened for T. gondii DNA using nested PCR. Eighteen (56.25%) samples were from females and fourteen (43.75%) from males. Toxoplasma gondii DNA was detected in 21 (65.6%) baboons. 13 females (40. 6%) and eight males (25%). Five baboons (24%) which tested positive were from the Institute of Primate Research colony but there was no statistical significance between them and the wild trapped (p=0.453). There was neither statistical significance (p=0.373) between sex and infection status nor between area of origin and infection status (p>0.05). These results indicate that olive baboons get infected with T. gondii in the wild and during captivity and may be significant reservoirs of human infections, especially where they may be trapped for bushmeat. We recommend a country-wide study to establish true prevalence of toxoplasmosis among non-human primates and identify associated parasite strains.

Cerebellar ataxia, neuronopathy, and vestibular areflexia syndrome (CANVAS) is caused by biallelic pathogenic expansions, or compound heterozygosity with other pathogenic variants in the RFC1 gene. CANVAS is estimated to be underdiagnosed, both because of the lack of formal diagnostic criteria and molecular challenges that translate to lesser access and high cost of routine testing. Our aim was to address the need for making CANVAS genetic testing routine, by designing a streamlined two-step PCR consisting of a short-allele screening PCR and a confirmatory PCR with fragment capillary electrophoresis detection. Exome sequencing of RFC1 was additionally foreseen to resolve potential compound heterozygosity cases. Specificity of our approach was evaluated using ataxia patients with known non-CANVAS diagnoses, and optimized using Southern blot confirmed CANVAS patients. We evaluated our approach by testing patients consecutively referred for clinically suspected CANVAS using first the two-step PCR, followed by exome sequencing. Our approach was able to accurately identify negative and confirm positive cases in prospectively collected suspected CANVAS patients presenting with at least three typical clinical signs. The proposed testing approach provides an alternative method able to clearly distinguish between CANVAS negative and positive cases and can be easily incorporated into the genetic diagnostic laboratory workflow.

Since certain Mycobacterium tuberculosis complex (MTBC) members, like M. bovis, are endemic in specific South African wildlife reserves and zoos, cases of clinically important nontuberculous mycobacteria (NTM) in wildlife may be neglected. Additionally, due to the inability of tests to differentiate between host responses to MTBC and NTM, the diagnosis of MTBC may be confounded by the presence of NTMs. This may hinder control efforts. These constraints highlight the need for enhanced rapid detection and differentiation methods for MTBC and NTM, especially in high MTBC burden areas. We evaluated the use of the GeneXpert MTB/RIF Ultra, the Hain CMdirect V1.0, and novel amplicon sequencing PCRs targeting the mycobacterial rpoB and ku gene targets, directly on antemortem respiratory samples from known MTBC-infected and NTM culture-positive African elephants (n=26 animals) and rhinoceros (n=23 animals). Our findings suggest that the Ultra is the most sensitive diagnostic test for MTBC DNA detection directly in raw antemortem respiratory specimens and that the rpoB PCR is ideal for Mycobacterium genus DNA detection and species identification through amplicon sequencing.

In recent research, a growing body of empirical evidence, suggests that prosocial impact at work can play a significant role in enhancing creativity and innovativeness. Drawing from Self Determination Theory, we hypothesized that basic psychological needs and benevolence satisfaction could serve as a mediating factor in the relation between employees perceived social impact and innovative work behavior and creativity, thus illuminating the manner in which contentment of psychological needs fosters inventive proclivities within the organizational milieu. Results from a study in Greece and Canada (N=528) showed that both perceived social impact and prosocial motivation are positively associated with innovative work behavior and creativity while autonomy and competence satisfaction mediate the relation between perceived social impact and the work outcomes examined within this study. Moreover, prosocial motivation was found to moderate the relation between benevolence satisfaction and innovativeness. Findings extend prior research on the role of prosociality on creative behavior at work and provide supporting evidence for the organizations that encourage and support employees’ initiatives to make a positive difference in the lives of others.

In the cone of positive quadratic forms C^{8 x 8}, it is shown that there exists in the neighbourhood of the quadratic form Q_{F_{22}} a large cluster of non-equivalent \Sigma_0-subcones of positive volume which are as well minimal as maximal.

Background. COVID-19 efforts were often ineffective in controlling the spread of the pandemic. Identifying ineffective controls during a pandemic is thus vital. Method. Utilizing publicly available data on COVID deaths in the counties of US states, we create an index to capture and interpret ineffectiveness in the efforts to reduce the spread of the pandemic in US counties. This index is based on the Intervened Poisson Distribution (IPD) introduced originally by Shanmugam. Motivation for the research idea occurred while we noticed the data dispersion of the COVID deaths is smaller than the average only in some counties. Under-dispersed data is common in statistical modeling. A novel approach we adapted in this article includes the estimation of an intervention parameter estimated through iterative non-linear optimization. Results. Twenty-five counties in California, Idaho, Minnesota, Mississippi, Montana, Nebraska, North Carolina, North Dakota, Texas, and Utah were found to be ineffective in controlling for fatalities based on the expected probability distribution. A review of the policies enacted in these areas would provide insight into ineffective prevention efforts, and some of these issues are documented in current literature. Conclusion. The IPD index an innovate way to document efficacy of interventions during pandemics. The IPD may identify ineffective efforts prior to statistical models intended to evaluate efficacy of efforts.

Background: Community pharmacies are well-placed to deliver well-being interventions; however, to date, nothing has been produced specifically for this setting. The aim of this study was to develop a positive psychology intervention suitable for a community pharmacy setting with the goal of increasing the well-being of community members. Methods: Intervention development consisted of three steps: Step 1- identify the evidence-base and well-being model to underpin the basis of the intervention (Version 1); Step 2 - model the intervention and gather user feedback to produce Version 2, and Step 3 - revisit the evidence-base and refine the intervention to produce Version 3. Results: Findings from nine studies (seven RCTs, one cross-sectional, one N-1 design plus user feedback were applied to model a 6-week ‘Prescribing Happiness (P-Hap)’ intervention, underpinned by the PERMA model plus four other components from the positive psychology literature (Three Good Things, Utilising Your Signature Strengths in New Ways, Best Possible Selves and Character Strengths). A PERMA-based diary was designed to be completed 3-days a week as part of the intervention. Conclusions: This work is an important development which will direct the future implementation of interventions to support well-being in this novel setting. The next stage is to gain the perspectives of external stakeholders on the feasibility of delivering the P-Hap for its adoption into community pharmacy services in the future.

Teacher burnout has been shown to be one of the most common negative consequences of the COVID-19 pandemic. This study aimed to identify distinct psychological resources and burnout risk profiles of teachers and examine their association with Kolb’s educator roles and their professional experience. Methods: The survey data were collected from 330 preschool and primary school (84 males, Mage = 38.3, SD = 9.14) teachers using a convenience sampling method. Results: The two-step cluster analysis revealed two distinct profiles. The first profile, “high psychological resources, no burnout risk,” was characterized by absent symptoms of burnout and increased levels of well-being, self-control, and positive emotionality. The second profile, “moderate psychological resources, mild burnout,” was associated with medium levels of well-being, self-control, and positive emotionality accompanied by mild burnout. Our findings highlighted that cluster one had a significantly higher score for the facilitator role and cluster two for the expert and coach roles. Additionally, teachers with less professional experience were more likely to belong to cluster one, considering their adequate skills in digital literacy. Conclusions: These findings provide new insights into the explanation of teacher burnout and the design of intervention programs.

Neoadjuvant systemic therapy has now become the the standard in early breast cancer management. Chemotherapy in combination with trastuzumab +/- pertuzumab targeted therapy can improve rates of pathologic complete response (pCR) in patients with HER2-positive breast cancer. Achieving a pCR is considered a good prognostic factor, in particular in patients with more aggressive breast cancer subtypes such as TNBC or HER2 positive cancers. Furthermore, most studies demonstrate that chemotherapy in combination with trastuzumab and pertuzumab is well tolerated. The retrospective analysis presented here concentrates on neoadjuvant therapy with the TCbH-P regimen, with a particular emphasis on patients over 60 years of age. We analysed the factors affecting the achievement of pCR and presented adverse effects of the applied therapies, which opened a discussion about optimizing the therapy of older patients with HER-2 positive breast cancer.

Part 1 of this 2-paper series identified a wide and deep network of context, generative mechanisms and outcomes responsible for psychosocial development in a performance basketball club. In this – part 2 – study, the stakeholder’s programme theories were tested during a full-season ethnography of the same club. The findings confirm the highly individualised nature of each young person’s journey. Methodologically, immersion in the day-to-day environment generated a fine-grain analysis of the processes involved, including: i) sustained attentional focus; ii) structured and unstructured skill building activities; iii) deliberate and incidental support; and iv) feelings indicating personal growth. Personal development in and through sport is thus shown to be conditional, multi-faceted, time-sensitive and idiosyncratic. The findings of this two-part study are considered to propose a model of psychosocial development in and through sport. This heuristic tool is presented to support sport psychologists, coaches, club administrators and parents to deliberately create and optimise developmental environments.

Long noncoding RNAs (lncRNAs) of virus origin accumulate in cells infected by many positive strand (+) RNA viruses to bolster viral infectivity. Their biogenesis mostly utilizes exoribonucleases of host cells that degrade viral genomic or subgenomic RNAs in the 5’-to-3’ direction until being stalled by well-defined RNA structures. Here we report a viral lncRNA that is produced by a novel replication-dependent mechanism. This lncRNA corresponds to the last 283 nucleotides of the turnip crinkle virus (TCV) genome, hence is designated tiny TCV subgenomic RNA (ttsgR). ttsgR accumulated to high levels in TCV-infected Nicotiana benthamiana cells when the TCV-encoded RNA-dependent RNA polymerase (RdRp), also known as p88, was overexpressed. Both (+) and (-) strand forms of ttsgR were produced in these cells in a manner dependent on the RdRp functionality. Strikingly, templates as short as ttsgR itself were sufficient to program ttsgR amplification, as long as the TCV-encoded replication proteins, p28 and p88, were provided in trans. Consistent with its replicational origin, ttsgR accumulation required a 5’ terminal G₃(A/U)₄ motif shown by others to be crucial for the replication of a TCV satellite RNA. More importantly, introducing a new G₃(A/U)₄ motif elsewhere in the TCV genome was alone sufficient to cause the emergence of another lncRNA. Collectively our results unveil a replication-dependent mechanism for the biogenesis of viral lncRNAs, thus suggesting that multiple mechanisms, individually or in combination, may be responsible for viral lncRNA production.

Business School rankings are “big business”, influencing donors and potential students alike, holding much sway over decanal and faculty priorities, particularly with respect to the curriculum as well as the focus and destination of research publications (i.e., in so called “top” journals). Over the past several years, the perverse effects of these priorities have begun to be acknowledged and new ratings and ranking systems have emerged. One promising new comer is the Positive Impact Rating (PIR), which uniquely and exclusively focuses on student perceptions of their business school’s priorities and the learning experience. In addition, it organizes schools by tier, in an effort to foster collaboration and continuous improvement, as opposed to ranked competition. If this new approach is to achieve its stated objective and help shift the focus of business schools to developing future business leaders and research output in alignment with a more sustainable world (and the United Nations Sustainable Development Goals), it is essential that the metrics used be – and perceived as - both valid and reliable. The current research aims to make a contribution in this regard, analyzing the results at one business school in detail and making recommendations for strengthening these aims. Results show that the parametric properties of the survey are highly interrelated suggesting that the predictive utility of the separate elements within scale could be improved. Additionally, biases in scores may exist dependent on where the responses are collected and who solicited them, as well as the students’ perception of their overall academic experience and on socio-cultural factors.

In this paper, by using the variational method, a sufficient condition for the unique existence of the stationary solution of the reaction-diffusion ecosystem is obtained, which directly leads to the global asymptotic stability of the unique equilibrium point. Besides, employing impulse control technique derives the globally exponential stability criterion of delayed feedback ecosystem.And numerical examples illuminate the effectiveness of impulse control, which has a certain enlightening effect on the actual epidemic prevention work . That is, in the face of the epidemic situation, taking a certain frequency of positive and effective epidemic prevention measures is conducive to the stability and control of the epidemic situation. particularly, the newly-obtained theorems quantifies this feasible step.

AbstractBaraton University dairy farm is an environment that attracts a microbiologist to inquire the composition of bacteria that exist there in. The knowledge of bacteria has in time and again amazed the life scientist community that have invested to acquire more information in this microbiology world.The study engages fundamental tests such as gram stain, endospore stain, and assays for specific microbial activities & enzymes, susceptibility on disinfectant and antibiotic, utilization of specific substrate and culture characteristics. The two organisms (gram negative and positive) tested positive for sucrose & lactose fermentation, Indole & Methly red, catalase & Oxidase, were both facultative and motile. On contrary, gram positive bacteria had spores and had a gamma haemolysis on Blood Agar, while gram negative bacteria haemolysed beta haemolysis. To draw a conclusion on the identity of the two organisms is that, the gram positive is a Bacillus ........ while gram negative is Escherichia coli.

Objectives: This study proposes a model that integrates three determinants of social cognitive theory (SCT) to explain the impact of FoMO on SNS usage. Design: A cross-sectional study is conducted using data for 259 participants recruited from a website. Main Outcome Measures: The analysis focuses on FoMO, social influence, positive outcome expectancy, refusal self-efficacy, and SNS-related behavior cloud-based sites. Data are examined using descriptive analysis and structural equation modeling. Results: The proposed model reported proper goodness of fit. FoMO does not directly or indirectly impact SNS usage through the determinants of SCT. However, social influence and refusal self-efficacy have a direct effect. Conclusions: The roles of the three determinants of SCT vary by stage of SNS usage. FoMO and refusal self-efficacy are more strongly related with SNS addiction. Further research, particularly longitudinal and intervention studies, is needed to examine the effects of specific factors on SNS addiction.

The present study investigates the reliability and validity of the Positive-Negative Relationship Quality (PN-RQ) scale in Turkey. In order to determine the validity and reliability of the scale, two studies were performed. The first was carried out with emerging adults (university students) who were in a romantic relationship (148 females, 43 males, and 2 unkown) and had the main purpose to examine the structure validity of the measurement tool in the Turkish sample with an exploratory analysis. Study 2 was performed online with 513 married adults (359 females, 149 males, and 5 unknown); confirmatory findings and criterion validity studies were added. Exploratory factor analyses revealed that relationship quality had a two-factor structure and that there was also a negative relationship between the factors. Confirmatory factor analyses on the married sample showed that the bi-factor model provided evidence for the multidimensional nature of the scale. Both studies demonstrated high internal consistency.

In this article, we establish the existence of and then compare smallest eigenvalues for nabla fractional boundary value problems involving a fractional difference boundary condition, using the theory of u0-positive operators with respect to a cone in a Banach space.

After reviewing foundational literature by Snyder, I will demonstrate how hope theory has been used in psychological interventions and psychiatry, how hope influences learning and medical outcomes, and how hope can be applied to difficult life circumstances. Using this literature review, hope theory will be interrogated. I will argue that hope theory has developed its way through the years to garner many strengths, however a few weaknesses do exist and will be described. I will conclude with recommendations for using hope theory in practice, including the medical field, psychological interventions and counseling, and student development.

From the perspective of individual resource and social capital, this paper aims to explain how employees’ political skill affect their job satisfaction and turnover intention, through the mediating role of popularity. Using a sample of 237 dyad surveys from supervisors and employees in the Yangtze River Delta of China, we found that: 1) political skill is positively correlated with job satisfaction and negatively correlated with turnover intention, and that these correlations are partially mediated by individual popularity; 2) positive affect (PA) moderates the relationship between political skill and job satisfaction, and negative affect (NA) moderates the relationship between political skill and turnover intention. The implications of our findings and future research directions are discussed.

Age-related diseases (ARDs) are the leading cause of death worldwide, and contribute to 90% of mortality in developed countries. Interestingly, the mortality rates of individual ARDs increase exponentially with age. Processes described by the exponential growth function typically involve a branching chain reaction or, more generally, a positive feedback loop. Here I propose that each ARD is mediated by one or several positive feedback loops (vicious cycles). I then identify critical vicious cycles in five major ARDs: atherosclerosis, hypertension, diabetes, Alzheimer’s and Parkinson’s. I also propose that the progression of ARDs can be halted by selectively interrupting the vicious cycles and suggest the most promising targets. An evolutionary perspective is also offered.

Rodents are known to be reservoirs of Toxoplasma gondii and keep the parasite circulation in the environment. We conducted biomonitoring to assess the role of sylvatic rodents in maintaining T. gondii and to analyse the prevalence and seroprevalence of the parasite in seven wild rodent species. Rodents were collected in our study sites (woodland and open grasslands) located in northeastern Poland and dissected. We collected brain, spleen, blood and serum samples. We applied both molecular (PCR assay, nested-PCR assay) and serological (ELISA and agglutination tests) methods to indicate the best approach for application in the biomonitoring of T. gondii in small mammals. We screened samples from 95 individuals sing PCR assays and found no T. gondii DNA. The agglutination test showed no signal. We found antibodies against T. gondii in 6 sera samples out of 74 analysed (seroprevalence = 8.11% [4.0-17.1]). Our results confirm that rodents participate in the life cycle of T. gondii as reservoirs of this parasite in the sylvatic environment. However, biomonitoring should be performed with the ELISA tests to search for T. gondii antigens, rather than a molecular approach only.

Verification of inclusivity of molecular detection methods gives indications on the reliability of viral infections diagnosis because of the tendency of viral pathogens to undergo sequence variation. This study was aimed to select inclusive probe based reverse transcription quantitative PCR (RT-qPCR) assays for the diagnosis of the most widespread and detrimental viruses infecting honeybees, namely the acute bee paralysis virus (ABPV), the black queen cell virus (BQCV), the chronic paralysis bee virus (CBPV), the deformed wing virus variants A (DWVA) and B (DWVB), and the sacbrood virus (SBV). Therefore, detection methods previously described were re-evaluated in silico for specificity and inclusivity. Based on this evaluation, selected methods were modified or new ones were designed and tested in duplex RT-qPCR reactions. The limits of detection (LODs), effect of multiplexing on sensitivity and viral RNA quantification potential in bees and hive debris were assessed. This study made available diagnostic assays able to detect an increased number of virus variants compared to previously described tests and two viral pathogens in a single PCR reaction.

The residual content of the only carbon source increases at the end of the exponential growth phase when P. putida BS3701 cells are cultured under nitrogen deficiency. Moreover, the mRNA quantity of some genes involved in carbon catabolism decreases. There are no available data on regulators capable of stimulating the target mRNA degradation in pseudomonads under nitrogen deficiency in the literature. We have applied both comparative genomics methods and a number of biomolecular ones to identify new small P. putida BS3701 RNAs. This is due to the fact that the formation of ncRNA-mRNA duplexes, recognized by RNases, is one of the main mechanisms for reducing the number of transcripts in a cell. We have used the following selection criteria: 1) the presence of the interest gene of the NtrC binding site and a sigma54-dependent promoter in the upstream area, 2) the conservative location of the interest gene in the P. putida and P. aeruginosa genomes, 3) increase in the product quantity of the interest gene in a cell under nitrogen deficiency on two different carbon sources (succinate and naphthalene), 4) the product detection of the interest gene in a fraction enriched with small RNAs. The expression of two new small RNAs has increased twice or more times under nitrogen deficiency on both succinate and naphthalene: expression of ng171 and ng379. ng379 contains narK RNA motif.

Outbreaks of coronavirus disease 2019 (COVID-19) have been recorded in different countries across the globe. The virus is highly contagious, hence early detection, isolation, and quarantine of infected patients will play an important role in containing the viral spread. Diagnosis in a mobile lab can aid to find infected patients in time. Here, we develop a field-deployable diagnostic workflow that can reliably detect COVID-19. Instruments used in this workflow could easily fit in a mobile cabin hospital and also be installed in the community. Different steps from sample inactivation to detection were optimized to find the fastest steps and portable instruments in detection of COVID-19. Each step was compared to that of the normal laboratory diagnosis set-up. From the results, our proposed workflow (80 min) was two times faster compared to that of the normal laboratory workflow (183 min) and a maximum of 32 samples could be detected at each run. Additionally, we showed that using 1% Rewocid WK-30 could inactivate the novel coronavirus directly without affecting the overall detection results. Comparison of our workflow using an in-house assay to that of a commercially acquired assay produced highly reliable results. From the 250 hospital samples tested, there was a high concordance 247/250 (98.8%) between the two assays. The in-house assay sensitivity and specificity were 116/116 (100%) and 131/134 (97.8%) compared to that of the commercial assay. Based on these results, we believe that our workflow is fast, reliable, adaptable and most importantly, field deployable.

We have used Daphnia cucullata as a model organism for the first time in the four deepest Latvian lakes from the Boreal biogeographical region lakes in order to find the genetic diversity of Daphnia cucullata populations. During the research, the most appropriate microsatellite markers for future genetic studies of Daphnia cucullata populations in Boreal biogeographical region lakes. It was the loci Dgm105 and Dgm101, in which the maximum number of alleles and the maximum number of private alleles were found. The Dgm105 locus had five private alleles (62% of all detected alleles), while the Dgm101 locus had four private alleles (57% of all detected alleles) in these loci. We was determined the observed heterozygosity (Hobs) and the expected heterozygosity (Hexp) level (by Hardy-Weinberg), the number of polymorphic loci, the number of detected alleles in each analyzed microsatellite locus, the average number of alleles at the locus (Na), the average effective number of alleles at the locus (Ne), FST of the population genetic differentiation, the genetic distance (D) (by Nei) and significance (χ2- test) of differences between the levels of observed and expected heterozygosity. It was shown that Daphnia cucullata populations from lakes with a low number of zooplankton taxa (Riča and Geraņimovas-Ilzas) have a higher genetic diversity compared to lakes with a high number of zooplankton taxa (Dridzis and Svente) in lakes. It was found, that Daphnia cucullata populations from lakes Dridzis and Svente have the least genetic distance and these populations form a single genetic group, as confirmed by clustering

Abstract: For the problem of small target of printed circuit board surface defects and low detection accuracy, the printed circuit board surface defect detection network DCR-YOLO is designed to meet the premise of real-time detection speed and effectively improve the detection accuracy. Firstly, the backbone feature extraction network DCR-backbone, consisting of two CR residual blocks and one common residual block, is used for small target defect extraction on printed circuit boards. Secondly, the SDDT-FPN feature fusion module is responsible for the fusion of high level features to low level features, while enhancing feature fusion for the feature fusion layer where the small target prediction head YOLO Head-P3 is located to further enhance the low level feature representation. the PCR module enhances the feature fusion mechanism between the backbone feature extraction network and the SDDT-FPN feature fusion module at different scales of feature layers. the C5ECA module is responsible for adaptive adjustment of feature weights and adaptive attention to the requirements of small target defect information, further enhancing the adaptive feature extraction capability of the feature fusion module. Finally, three YOLO-Heads are responsible for predicting small target defects for different scales. Experiments show that the DCR-YOLO network model detection Map reaches 98.58%, the model size is 7.73MB, which meets the lightweight requirement, and the detection speed reaches 103.15fps, which meets the application requirements for real-time detection of small target defects.

The device and method in accordance with the constant pressure regulation of micro droplet PCR in microfluidic chips are developed to explore the optimization way for the micro droplet movement, fragmentation, and bubble generation in microfluidic chips in existing digital PCR technology. In the developed device, an air source device is adopted to regulate the pressure in the chip, such that micro droplet generation and PCR amplification without bubbles can be achieved. In 3 min, the sample in 20ul will be distributed into nearly 50,000 water-in-oil droplets exhibiting a diameter of about 87μm, and the micro droplet will be subjected to a close arrangement in the chip without air bubbles. The device and chip are adopted to quantitatively detect the human gene . As indicated by the experimental results, a good linear relationship exists between detection signal and DNA concentration ranging from 101~ 105 copies /μL (R2=0.999). The micro droplet PCR devices based on constant pressure regulation chips exhibit a wide variety of advantages (e.g., achieving high pollution resistance, micro droplet fragmentation and integration avoidance, reducing human interference, and standardizing results). Thus, micro droplet PCR devices based on constant pressure regulation chips have promising applications in nucleic acid quantification.

Leaf color mutants (LCMs) are important resources for studying diverse metabolic processes such as chloroplast biogenesis and differentiation, pigments’ biosynthesis and accumulation, and photosynthesis. However, in Dendrobium officinale, LCMs are yet to be fully studied and exploited due to the unavailability of reliable RGs (reference genes) for qRT-PCR (quantitative real-time reverse transcription PCR) normalization. Hence, this study took advantage of previously released transcriptome data to select and evaluate the suitability of ten candidate RGs, including Actin (Actin), polyubiquitin (UBQ), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1-α (EF1α), β-tubulin (β-TUB), α-tubulin (α-TUB), 60S ribosomal protein L13-1 (RPL13AD), aquaporin PIP1-2 (PIP1-2), Intima protein (ALB3) and Cyclin (CYCB1-2) for normalizing leaf color-related genes’ expression levels via qRT-PCR. Stability rankings analysis via common software Best-Keeper, GeNorm, and NormFinder disclosed that both ten genes met the requirements of RGs. Of them, EF1α exhibited the highest stability and was selected as the most reliable. The reliability and accuracy of EF1α were confirmed through qRT-PCR analysis of fifteen chlorophyll pathway-related genes. The expression patterns of these genes via EF1α normalization were consistent with the results by RNA-Seq. Our results will pave the way for molecular dissection of leaf color mutations in D. officinale.

Abstract: In 2016, we conducted an entomological survey in a railway transect between Banfora and Ouagadougou, Burkina Faso. The aim was to evaluate the risk factors for arbovirus epidemics, including vector infection status, in areas representative of the country. Aedes aegypti mosquitoes were collected at larval stage from four study sites and reared until adult stage and kept in RNAlater for detection of arbovirus RNA. In the laboratory, the mosquito specimens were screened for dengue virus (DENV) and chikungunya virus (CHIKV) using one step real-time qRT-PCR. We detected one DENV-2 positive pool from Ouagadougou, giving a minimum infec-tion rate (MIR) of 16.67, and 6 CHIKV positive pools, giving a MIR of 66.67 from Ouagadougou, but also in Banfora and Boromo. The qRT-PCR is a useful tool for the surveillance of arboviruses of public health importance in Burkina Faso and may be incorporated into disease surveillance and control programs in Burkina Faso.

Soybean (Glycine max L.) is an important crop that serves as a source of edible oil and protein. However, little is known about its molecular mechanism of adaptation to extreme environmental conditions. Based on the Arabidopsis thaliana sequence database and Phytozome, a soybean gene that was highly homogenous with the reduced induction of the non-photochemical quenching2 (AtRIQ2) gene, GmRIQ2-like (accession NO.: Glyma.04G174400), was identified in this study. The gene structure analysis revealed that GmRIQ2-like encoded a transmembrane protein. Elements of the promoter analysis indicated that GmRIQ2-like participated in the photosynthesis and abiotic stress pathways. The subcellular localization results revealed that the protein encoded by GmRIQ2-like was located in chloroplasts. The quantitative real-time (qRT)-PCR results revealed that GmRIQ2-like-overexpression (OE) and -knock-out (KO) transgenic soybean seedlings were cultivated successfully. The relative chlorophyll (Chl) and zeaxanthin contents and Chl fluorescence kinetic parameters demonstrated that GmRIQ2-like dissipated excess light energy by enhancing the non-photochemical quenching (NPQ) and reduced plant photoinhibition. These results suggested that GmRIQ2-like was induced in response to strong light and depressed Chl production involved in soybean stress tolerance. These findings indicate that the transgenic seedlings of GmRIQ2-like could be used to enhance strong light stress tolerance and protect soybean plants from photoinhibition damage. This study will serve as a reference for studying crop photoprotection regulation mechanisms and benefits the research and development of new cultivars.