The aim of this study was to investigate the differentially expressed genes associated with intramuscular fat deposition in the longissimus dorsi muscle of Xinjiang Brown Bulls. The longissimus dorsi muscles of 42 Xinjiang Brown Bulls were selected under the same feeding conditions. The intramuscular fat content of muscle samples were determined by Soxhlet extraction method, from which 5 samples with high intramuscular fat content (HIMF group) and 5 samples with low intramuscular fat content (LIMF group) were selected. It was found that the intramuscular fat content of HIMF group was 46.054% higher than that of LIMF group. Muscle samples produced by paraffin section for morphological observation. It was found that the fat richness of the HIMF group was better than that of the LIMF group. Transcriptome sequencing technology was used to analyze the gene expression differences of longissimus dorsi muscle. Through in-depth analysis of the longissimus dorsi muscle by transcriptome sequencing technology, we screened a total of 165 differentially expressed genes. The results of Gene Ontology (GO) enrichment analysis showed that the differentially expressed genes in the two groups were mainly clustered in biological pathways related to carbohydrate metabolic processes, redox processes and oxidoreductase activities. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the differentially expressed genes were significantly clustered in 15 metabolic pathways, which mainly covered fatty acid metabolism (related to lipid metabolism and glucose metabolism), pentose phosphate pathway, Peroxisome Proliferator-Activated Receptors (PPAR) signalling pathway and other important metabolic processes. The three genes that were predominantly enriched in the glycolipid metabolic pathway by analysis were SCD5, CPT1C, and FBP2, all of which directly or indirectly affect intramuscular fat deposition. In summary, the present study investigated the differences in gene expression between high and low intramuscular fat content in the longissimus dorsi muscle of Xinjiang Brown Bulls by transcriptome sequencing technology, and revealed the related signalling pathways. Therefore, we hypothesized that SCD5, CPT1C, and FBP2 were the key genes responsible for the significant differences in intramuscular fat content of the longissimus dorsi muscles in a population of Xinjiang Brown Bulls. We expect that these findings will provide fundamental support for subsequent studies exploring key genes affecting fat deposition characteristics in Xinjiang Brown Bulls.

Chronic stress is a pervasive issue that significantly impacts both physical and mental health. Unlike acute stress, which is short-lived and can be beneficial, chronic stress persists over extended periods, leading to a range of adverse health outcomes. This paper explores the myriad physical, mental, and behavioral consequences of chronic stress, emphasizing the underlying physiological mechanisms and potential long-term health implications. Through a comprehensive literature review and data analysis utilizing Python, we aim to provide an in-depth understanding of chronic stress and its effects. Key findings include the critical roles of cortisol and other stress-related chemicals in exacerbating health issues, and the potential for the normalization of high stress levels as an adaptive yet harmful response.

Background/Objectives: The latest WHO classification underscores the importance of the complete blood count (CBC) and cell morphology assessment in diagnosing lymphoid neoplasms. This study evaluated the diagnostic role of Research Use Only (RUO) parameters from the BC-6800 Plus Mindray analyzer in differentiating cell chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), and other lymphoproliferative disorders. Methods: CBCs were performed on 90 patients (66% male, median age 67 years) with pathological blood counts (ALL, n=14; CLL, n=47; other lymphoproliferative disorders, n=29) at Novara’s University Hospital. Univariable and multivariable logistic regression assessed the association of basic (WBC, Hb, RDW, NE#, LY#, MO#, PLT) and research cell parameters (NLR, NMR, LMR, NeuX, NeuY, NeuZ, LymX, LymY, LymZ, MonX, MonY, MonZ). Results: In multivariable analysis, Hb (p=0.02), NeuY (p=0.04), and MonY (p=0.01) independently predicted CLL versus ALL, correctly classifying 93.4% of cases (AUC 0.91, 95% CI 0.81-1.0). MO# (p=0.003), LymY (p<0.0001), and MonY (p=0.004) were independent predictors of CLL versus other lymphoproliferative disorders, classifying 77.6% of cases (AUC 0.86, 95% CI 0.78-0.95). NeuZ (p=0.01) and NeuY (p=0.04) independently predicted ALL versus other lymphoproliferative disorders, classifying 93% of cases (AUC 0.98, 95% CI 0.95-1.0). Conclusions: The use of RUO parameters offers valuable, cost-effective support in the early diagnosis of ALL, CLL, and other lymphoproliferative disorders.

Cholera is a diarrheal disease prevalent in populations without access to clean water. Cholera is caused by Vibrio cholerae, which colonizes the upper small intestine in humans once ingested. A growing number of studies suggests the gut microbiome composition modulates animal behavior. Zebrafish are an established cholera model that can maintain a complex, mature gut microbiome during infection. Larval zebrafish, which have immature gut microbiomes, provide the advantage of high-throughput analyses for established behavioral models. We identified the effects of V. cholerae O1 El Tor C6706 colonization at 5 days post-fertilization (dpf) on larval zebrafish behavior by tracking startle responses at 10 dpf. We also characterized the larval gut microbiome using 16S rRNA sequencing. V. cholerae-infected or uninfected control groups were exposed to either an alternating light/dark stimuli or a single-tap stimulus, and average distance and velocity were tracked. While there was no significant difference in the light/dark trial, we report a significant decrease in distance moved for C6706-colonized larvae during the single-tap trial. This suggests that early V. cholerae colonization of the larval gut microbiome has a dampening effect on sensorimotor function, supporting the idea of a link between the gut microbiome and behavior.

The clinical application of collagen-based biomaterials is expanding rapidly, especially in tissue engineering and cosmetics. While oral supplements and injectable skin boosters are popular for enhancing skin health, clinical evidence supporting their effectiveness remains limited. Injectable products show potential in revitalizing skin, but safety concerns persist due to challenges in sterilization and the risk of biological contamination. Traditional methods of sterilization (heat and irradiation) can denature collagen. This study addresses these issues by introducing a novel technique: double filtration and low-temperature steam sterilization of a collagen gel. In vitro tests document sterility and confirm that the collagen structure remained intact without modification. An in vivo study found no adverse effects in rats or significant lesions at the implantation site over 13 weeks. These results suggest that this novel method to process collagen gels is a safe and effective skin booster. Advanced processing methods are likely to mitigate safety risks associated with injectable collagen products, though further research is needed to validate their biological effectiveness and clinical benefits.

The role of astro- and microglial cells in the pathogenesis of epilepsy is currently under active in-vestigation. It is proposed that the activity of these cells may be regulated by agonists of peroxi-some proliferator-activated nuclear receptors (PPARs). The objective of this study was to investi-gate the effects of the PPAR β/δ agonist GW0742 (Fitorine) on the behavior and gene expression of astro- and microglial proteins involved in the regulation of epileptogenesis in rat brain in a lithi-um-pilocarpine model of temporal lobe epilepsy (TLE). A seven-day administration of GW0742 at a dose of 5 mg/kg/day has been shown to suppress TLE-related behavioral hyperactivity in the Open Field test and to attenuate the TLE-induced increase in gene expression and protein levels of astro- and microglial activation markers Gfap and Aif1 in the dorsal hippocampus and temporal cortex. Furthermore, GW0742 reversed the TLE-related increase in Lcn2 gene expression (a marker of the A1 phenotype of astrocytes) in the temporal cortex, the increase in the ratio of Nos2/Arg1 gene expression (markers of the M1/M2 phenotypes of microglia), and the decrease in Arg1 gene ex-pression in the dorsal hippocampus. The results suggest that GW0742 may mitigate behavioral and biochemical abnormalities in a lithium-pilocarpine model of TLE.

Diabetic Foot Ulcer (DFU) is a severe complication of diabetes mellitus (DM), often leading to hospitalization and non-traumatic amputations in the United States. It is particularly prevalent among individuals of Hispanic descent, with a prevalence rate exceeding 10% [1, 2]. The target of this study is to identify markers of severity of DFU. This work follows a strategy that uses first Electronic Health Records and machine learning to identify risk factors reported as measurements of blood tests, such as cholesterol, blood sugar, and specific proteins in a cohort of thousands of patients. After this RNA risk factors are predicted using samples of bulk and single-cell gene expression across patients with different levels of DFU severity. We found the Albu-min/Creatinine Ratio (ACR) test as a key factor in severe DFUs. We also found a cluster of cells that harbor high expression of Apolipoprotein E protein (APOE) and it is significant in non-healing DFU cases. Overall, the study shows how the use of hundreds of thousands of EHR can reduce and inform the search space of molecules in a few samples of bulk-single cell transcriptomics and identify molecular markers of severity of DFU.

Mulberries is a type of fruit tree that is rich in nutrients and possesses both medicinal and dietary properties, deeply loved by consumers. The fragrance is a significant factor in the sensory quality of mulberries, playing a crucial role in consumer purchasing behavior. 'Baichang' mulberry is a unique variety of mulberry that emits a strong, creamy-like fragrance, which has enormous potential market value. However, the key metabolites and mechanisms responsible for the formation of this fragrance are still unclear. In this study, we utilized GC-MS technology and RNA-seq to examine the Volatile Organic Compounds (VOCs) and gene expression in 'Baichang' (Morus macroura) mulberries and 'Hongchang' (Morus laevigata) mulberries, the latter of which do not have a distinct fragrance. The results showed that 661 volatile metabolites were detected in both mulberry varieties. Most of these were found in higher relative amounts in the 'Baichang' mulberry variety, with 47 types being present only in the 'Baichang' mulberries, suggesting a rich composition of VOCs in the 'Baichang' mulberry. 312 metabolites were identified as differential metabolites; combining the sensory annotation information and relative content of VOCs, 70 candidate VOCs were annotated as related to the formation of 'sweet, waxy, coconut, and oily' sensations. They could potentially be the key metabolites contributing to the unique aroma of 'Baichang' mulberries. The relative odor activity value (rOAV) analysis revealed that 2(3H)-Furanone, 5-hexyldihydro- was the key VOC contributing to the formation of the special aroma of 'Baichang' mulberries. By RNA-seq, 19 key structural genes related to ester synthesis were identified, including AAT , ACX , ALDH, CYP, EHL, FAD, HPL, and LOX. The high expression of these structural genes in the "Baichang" mulberry fruit is the main reason for its unique creamy aroma. This study provides insights into the aroma differences among different kinds of mulberrie.

Lactococcus lactis is a potential bacterial cell factory to develop delivery systems for vaccines and therapeutic proteins. Large progress has been made in applications using engineered L. lactis against, e.g., inflammatory bowel disease and cervical cancer, but improvement of secretion and cell anchoring efficacy is still desirable. A double-labelling method based on biarsenical hairpin binding and nickel-polyhistidine affinity was used for visualization of protein trafficking and quantification of targeted proteins on the cell surface and in the cytoplasm. To investigate the importance of mature domain targeting signals (MTSs), we generated truncated constructs encoding 126, 66, and 26 amino acid residues from the N-terminus of the basic membrane protein A (BmpA) and fused those with the gene for the human papillomavirus serotype 16 (HPV16) E7 oncoprotein. Overexpression of fusion proteins was observed to come at a cost of cell proliferation. L. lactis cells produced and displayed the shortest fusion protein only with difficulty, suggesting that the entire absence of a homologous sequence containing MTSs significantly impedes export and surface anchoring of fusion proteins. With 40 amino acids following the signal peptide and containing one MTS, effective translocation was possible. Mutations of MTSs towards increased hydrophobicity resulted in an increase of secreted and surface-displayed fusion protein, suggesting the potential to design rationally improved constructs.

The aim of this study was to investigate the differentially expressed genes associated with intramuscular fat deposition in the longissimus dorsi muscle of Xinjiang Brown Bulls. The longissimus dorsi muscles of 42 Xinjiang Brown Bulls were selected under the same feeding conditions. The intramuscular fat content of muscle samples were determined by Soxhlet extraction method, from which 5 samples with high intramuscular fat content (HIMF group) and 5 samples with low intramuscular fat content (LIMF group) were selected. It was found that the intramuscular fat content of HIMF group was 46.054% higher than that of LIMF group. Muscle samples produced by paraffin section for morphological observation. It was found that the fat richness of the HIMF group was better than that of the LIMF group. Transcriptome sequencing technology was used to analyze the gene expression differences of longissimus dorsi muscle. Through in-depth analysis of the longissimus dorsi muscle by transcriptome sequencing technology, we screened a total of 165 differentially expressed genes. The results of Gene Ontology (GO) enrichment analysis showed that the differentially expressed genes in the two groups were mainly clustered in biological pathways related to carbohydrate metabolic processes, redox processes and oxidoreductase activities. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the differentially expressed genes were significantly clustered in 15 metabolic pathways, which mainly covered fatty acid metabolism (related to lipid metabolism and glucose metabolism), pentose phosphate pathway, Peroxisome Proliferator-Activated Receptors (PPAR) signalling pathway and other important metabolic processes. The three genes that were predominantly enriched in the glycolipid metabolic pathway by analysis were SCD5, CPT1C, and FBP2, all of which directly or indirectly affect intramuscular fat deposition. In summary, the present study investigated the differences in gene expression between high and low intramuscular fat content in the longissimus dorsi muscle of Xinjiang Brown Bulls by transcriptome sequencing technology, and revealed the related signalling pathways. Therefore, we hypothesized that SCD5, CPT1C, and FBP2 were the key genes responsible for the significant differences in intramuscular fat content of the longissimus dorsi muscles in a population of Xinjiang Brown Bulls. We expect that these findings will provide fundamental support for subsequent studies exploring key genes affecting fat deposition characteristics in Xinjiang Brown Bulls.

Importance: Neuropsychiatric disorders like schizophrenia and bipolar disorder lack objective diagnostic markers, hindering accurate diagnosis and treatment. A novel approach using patient-derived cerebral organoids and digital analysis of electrophysiological recordings could provide much-needed objective biomarkers.Objective: To develop a digital analysis pipeline that can identify distinct electrophysiological signatures of schizophrenia and bipolar disorder using multi-electrode array recordings from patient-derived cerebral organoids.Design: This was an experimental study using previously published data. The study type can be classified as a case-control study, comparing cerebral organoids and 2D neuronal cultures derived from patients with schizophrenia, bipolar disorder, and healthy controls. The study utilized multi-electrode array recordings for analysis. Setting: The study was conducted in a laboratory setting using previously recorded data. Participants: The study included a total of 24 Caucasian subjects, comprising three groups: patients with schizophrenia, patients with bipolar disorder, and healthy controls. The specific number of participants in each group is not provided. The participant pool consisted of 17 males and 7 females, with an average age of 38.8 years.Main Outcomes and Measures: The primary outcome was the accuracy of a support vector machine classifier in distinguishing between healthy control, schizophrenia, and bipolar disorder samples based on electrophysiological features extracted from the recordings. Key cellular-digital biomarkers were identified using minimum redundancy maximum relevance feature selection.Results: This study included 24 Caucasian subjects (17 males, 7 females; mean age 38.8 years) with schizophrenia (SCZ), bipolar disorder (BPD), and healthy controls. Our Support Vector Machine classifier achieved 95.8% accuracy in distinguishing SCZ from control samples in 2D neuronal cultures under both baseline and post-electrical stimulation (PES) conditions. For cerebral organoids, classification accuracy was 83.3% under baseline conditions, improving to 91.6% under PES when distinguishing between control, SCZ, and BPD samples. Key features for classification included channel-specific measures such as median, covariance, autocorrelation, and kurtosis of neural activity. Confusion matrices visualize classification performance, with most misclassifications occurring in the BPD class under baseline conditions. These results demonstrate that our digital analysis pipeline can effectively distinguish between healthy control and patient-derived samples using electrophysiological features from multi-electrode array recordings, particularly under PES conditions.Conclusions and Relevance: This digital analysis pipeline demonstrates the potential to identify objective electrophysiological signatures of schizophrenia and bipolar disorder using patient-derived cerebral organoids. This approach could lead to improved diagnostic accuracy and personalized treatment strategies for neuropsychiatric disorders.

Introduction: The environmentally acquired aerobic spore-forming (EAS-Fs) bacteria that are ubiquitous in nature (e.g. soil) are transient colonisers of the mammalian gastro-intestinal tract. Without regular exposure, their numbers quickly diminish. These species of bacteria have been suggested to be essential to the normal functioning of metabolic and immunogenic health. The modern Western lifestyle restricts exposure to these EAS-Fs, possibly explaining part of the pathogenesis of many Western diseases. To date, the only animal studies that address specific microbiome modelling are based around germ-free animals. We have designed a new animal model that specifically restricts exposure to environmental sources of bacteria. Methodology: A new protocol, termed Super Clean, that involves housing mice in autoclaved individually ventilated cages (IVCs), with autoclaved food / water, and strict ascetic handling practice was first experimentally validated. The quantification of EAS-Fs was assessed by heat-treating faecal samples and measuring colony forming units (CFUs). This was then compared to mice in standard-conditions. Mice were housed in their respective groups from birth until 18 months. Stool samples were taken throughout the experiment to assess for abundance in transiently acquired environmental bacteria. Clinical, biochemical, histological, and gene expression markers were analysed for diabetes, hypercholesterolaemia, obesity, inflammatory bowel disease, and non-alcoholic fatty liver disease (the “diseases of the West”). Results: Our results show that stringent adherence to the Super Clean protocol produces a significantly decreased abundance of aerobic spore-forming Bacillota after 21 days. This microbiomic shift was correlated with significantly increased levels of obesity and impaired glucose metabolism. There was no evidence of colitis, liver disease or hypercholesterolaemia. Conclusion: This new murine model successfully isolates EAS-Fs, and has the potential utility for future research allowing for the investigation into the clinical impact living in relative hygienic conditions.

The Galápagos archipelago is a vast reservoir of terrestrial and marine biodiversity, owing in large part to its relatively recent volcanic genesis and colonization by humans. This unique ecological system is particularly susceptible to human, animal, and environmental impacts. Climate change, globalization, and the blurring of human-domestic animal-wildlife interfaces are poised to bring new threats and challenges to the region. A One Health perspective that simultaneously considers human, animal, and environmental health is imperative in assessing and mitigating the challenges facing the Galápagos Islands.In Part I of this review, we provide the historical context for biodiversity in the archipelago; discuss the role of invasive species in habitat destruction, fragmentation, and competition with endemic species; and summarize the established and emerging infectious disease threats. We also discuss the imperative to implement research, surveillance, and preventative measures to identify and manage future threats from a One Health perspective, with a specific emphasis on implications for wildlife health. In Part II, we outline the socioeconomic context of life in the Galápagos Islands, evaluate the current and predicted effects of climate change, and discuss direct anthropogenic factors affecting Galápagos biodiversity, such as tourism, fishing, pollution, and the illegal wildlife trade. We also examine the impact of the COVID-19 pandemic in the region. Throughout this two-part review, we build a cohesive picture of One Health in the Galápagos Islands by integrating past work, current needs, and emerging threats. We also consider overarching goals for conservation, ecosystem management, and socioeconomic sustainability that have been previously defined by both governmental and non-governmental stakeholders, and we identify discrete, implementable, and interdisciplinary recommendations that will facilitate achievement of those goals.

Adhesive-invasive E. coli have been suggested to be associated with the development of Crohn's disease (CD). It is assumed that they can provoke the onset of the inflammatory process as a result of the invasion of intestinal epithelial cells, and then, due to survival inside macrophages, stimulate chronic inflammation and the development of granuloma. In previous reports, we have shown that passage of the СD isolate ZvL2 on minimal medium M9 supplemented with sodium propionate (PA) as a carbon source stimulates, and with glucose - inhibits the adherent-invasive properties and the ability to survive in macrophages. This effect was reversible and not observed for the laboratory strain K12 MG1655. We were able to compare the isogenic strain AIEС in two phenotypes - virulent (ZvL2-PA) and non-virulent (ZvL2-GLU). We showed that, unlike ZvL2-GLU, the ZvL2-PA activates the production of ROS and cytokines when interacting with neutrophils. The laboratory strain does not cause a similar effect. Differences in neutrophil NADH-oxidase activation and ζ potential for ZvL2-GLU and ZvL2-PA are associated with changes in the abundance of membrane proteins. Using differential 2D electrophoresis and LC-MS, we identified membrane proteins that may be associated with the activation of the virulence properties of the СD isolate ZvL2.

Regulations and guidelines on water quality are used according to physicochemical but scarcely biological variables to protect ecosystems and control water use. This study innovates using in-sects as bioindicators to establish the health status of an aquatic ecosystem in the Latin American's lentic bodies. Water quality in urban lentic bodies in the Metropolitan Region, Chile, was evaluated from of aquatic insect assemblages and physicochemical variables for conserving aquatic life. Evaluations were carried out in parallel in four sampling stations in three water bodies, two seasons, and 2–3-year series, with three replicates. Aquatic insects were randomly sampled and identified by families; abundance, and richness differences were compared with non-parametric tests. Family Biotic Indices (FBI) were calculated. Physicochemical variables were measured using portable multiparametric and laboratory chemical analysis to determine water quality. A new physicochemical variable (total suspended solids) was established, which had a greater incidence with the results obtained for the FBI. Based on the nitrogen and phosphorus levels in the water bodies, all of them were eutrophics. Given the ecosystem diversity and complexity, studies should delve deeper into wetlands to establish methods that contribute to determining water quality, using insects as bioindicators and physicochemical variables.

Carbamate insecticides have been used for over four decades to control brown planthopper, Nilaparvata lugens, but resistance has been reported in many countries, including Korea. The test results on resistance to fenobucarb showed that the LC₅₀ values were 3.08 for the susceptible strain, 10.61 for the 2015 population, and 73.98 for the 2019 population. Compared to the susceptible strain, the 2015 and 2019 populations exhibited resistance levels 3.4 and 24.2 times higher, respectively. To elucidate the reason for the varying levels of resistance to fenobucarb in these populations, mutations in the ace1 gene, the target site of carbamate, were investigated, but no previously reported mutations were confirmed. Through RNAseq analysis focusing on the expression of detoxification enzyme genes as an alternative resistance mechanism, it was found that the carboxylesterase gene Nl-EST1 was overexpressed 2.4 times in the 2015 population and 4.7 times in the 2019 population compared to the susceptible strain. This indicates a strong correlation between the level of resistance development in each population and the expression level of Nl-EST1. Previously, Nl-EST1 was reported in an organophosphorus insecticide-resistant strain of Sri Lanka 2000. Thus, Nl-EST1 is crucial for developing resistance to organophosphorus and carbamate insecticides. Resistance-related genes such as Nl-EST1 could serve as expression markers for resistance diagnosis, and it can apply to integrated resistance management of N. lugens.

Protected and conserved areas are a key area-based strategy to address the biodiversity and climate change crises. Fundamental to understand their performance over time are indicators. The Digital Observatory for Protected Areas (DOPA) was born in 2013 as a set of open access web services and applications to be used to assess, monitor, and report on protected and conserved areas. For over a decade it has delivered over 50 indicators to support policy processes, national and regional governments, and practitioners. DOPA has also developed a versatile and efficient back-end approach that is widely applicable in other area-based conservation contexts. Here, we describe the methods and workflows behind DOPA´s back end and provide examples of policy relevant questions it can answer. We discuss key advantages and limitations of this systematic and replicable approach and explore the use of this back-end architecture to inform progress in area-based conservation targets for the following decades. This approach, embedded in multiple services provided by the Knowledge Centre for Biodiversity of the European Commission (KCBD), can also support the implementation and monitoring of area-based targets of the Kunming Montreal Global Biodiversity Framework at international, regional, and national levels.

The study´s goals were to determine the health status of a group of heart transplant recipients (HTR) and their level of physical activity and to compare the health status among them and with a group of healthy sedentary individuals. Fifty-four HTR and eighteen S were assigned to four groups, according to their level of physical activity (determined with The International Physical Activity Questionnaire); patients with a low, a moderate and a high level of physical activity (HTRL, HTRM and HTRH, respectively) and sedentary individuals (S). Participants underwent a basic blood analysis and several tests to assess their cardiovascular, neuromuscular, functional mobility condition and their quality of life. The S and HTRH were very similar in terms of BP, HR and blood analysis while HTRM and HTRL differed from both S and HTRH in these parameters. Regarding the cardiovascular, neuromuscular, functional mobility, and quality of life variables assessed in this study, HTRH showed the best results across all of them, followed by S, HTRM, and HTRL. It is suggested that the weekly level of physical activity of HTR should be high, what might help them to enhance their health and quality of life.

This study provides a comprehensive analysis of the conformational dynamics of pseudo-endocannabinoid receptors (p-CBRs) CB₁ and CB₂ in response to ligand binding. The ligands studied include N-arachidonoyl dopamine (NADA), Cannabidiol (CBD), known agonists (FUB-MDMB and E3R), and inverse agonists (Taranabant and AM630). Using homology models based on crystal structures (PDB ID: 5U09 for CB₁ and 5ZTY for CB₂), molecular docking, and molecular dynamics (MD) simulations, the research validates the structural integrity and dynamic behaviours of these pseudo-receptors. Induced-fit docking revealed NADA as having the highest binding affinity for both receptors, with docking scores of −13.92 kcal/mol for CB₁ and −13.32 kcal/mol for CB₂, facilitated by key hydrogen bond interactions. MD simulations over 250 ns at ~298 K showed successful equilibration, indicating the most stable interaction for NADA, thereby further validating the models. CB₁ and CB₂ exhibited distinct conformational responses to ligands, with Cα RMSD and Cα RMSF analyses suggesting a more rigid binding pocket for CB₁ and a flexible binding pocket for CB₂. These findings highlight the significance of receptor behaviour and dynamics in the development of cannabinoid-based therapeutics, supporting the potential of CBD and NADA as modulators for CB₁ and CB₂, respectively.

The main objective of our study was to understand the rheological behavior of various types of cheeses with mold subjected to multiple stresses during processing/handling, transport, or storage, aiming to maintain or even improve product quality, using computer-assisted penetration methods, and advanced regression algorithms. Uniaxial penetration tests with a cone at a constant speed were conducted using a universal Hounsfield HK1S testing equipment connected to a computer to analyze the texture behavior (tangential stress, flow index, apparent viscosity) of four of the most common types of cheese with mold, depending on the cone's tip angle (9, 19, and 90°) and penetration speed (12, 30, and 60 mm/min). From the results obtained for the four categories of mold cheeses (Brie, Camembert, Dorblue, or Roquefort), the amplitude of the speed and angle of penetration were considered as the main influencing factors for the shear deformation (τc), flow index (Kf), and apparent viscosity (ηa). The moisture and firmness of the mold cheeses are closely linked and depend on the type of mold, the maturation process, or storage conditions. Careful control of these factors is essential to achieve the desired texture and taste characteristics in cheeses with mold.

Listeria monocytogenes is an emerging pathogen responsible for the serious foodborne disease, listeriosis. The commensal gut microbiota is the first line of defense against pathogen internalization. The gut microbiome can be modified by prebiotic substrates, which are frequently added to food products and dietary supplements. Prebiotics should selectively support the growth of beneficial microbes and thus improve host health. Nevertheless, little is known about their effect on the growth of L. monocytogenes. The aim of this study was to evaluate the growth ability of four L. monocytogenes strains, representing the most common serotypes, on prebiotic oligosaccharides (beta-(1,3)-D-glucan, inulin, fructooligosaccharides, galactooligosaccharides, lactulose, raffinose, stachyose, and 2´-fucosyllactose and a mixture of human milk oligosaccharides) as a sole carbon source. The results showed that only beta-(1,3)-D-glucan was metabolized by L. monocytogenes. Therefore, the safety of beta-(1,3)-D-glucan with respect to listeriosis development should be further studied.

The Anterior Cruciate Ligament (ACL) is a crucial stabilizing ligament in the knee, essential for maintaining knee joint stability and proper biomechanical function. ACL rupture is one of the most common injuries, especially prevalent among athletes participating in high-demand sports that involve pivoting, cutting, and jumping. Such injuries not only lead to immediate functional impairment but also predispose individuals to long-term consequences, including osteoarthritis if left untreated. ACL reconstruction using a graft, whether autograft or allograft, has become the gold standard treatment method aimed at restoring knee stability and function. However, the postoperative healing process of the graft within the bone tunnel presents significant challenges due to its complexity. This healing involves a coordinated sequence of molecular and cellular events, including inflammation, proliferation, and remodeling phases. These phases are governed by a myriad of molecular signals, growth factors, and cellular interactions that ultimately determine the success of the graft integration and the restoration of ligamentous function. The review article "Molecular Biology of ACL Graft Healing: Early Mechanical Loading Perspective" delves deeply into these intricate biological processes. It provides a comprehensive analysis of the underlying molecular mechanisms that facilitate graft healing. Furthermore, it emphasizes the critical role of early mechanical loading in this context. Mechanical loading, when applied appropriately, has been shown to positively influence graft healing by enhancing cellular responses, collagen fiber alignment, and overall biomechanical properties of the graft. By exploring the impact of early mechanical loading, the review sheds light on how mechanical stimuli can be optimized to improve clinical outcomes. This understanding is vital for developing effective rehabilitation protocols that promote faster and more robust graft healing. The article offers valuable insights for clinicians and researchers, aiming to bridge the gap between molecular biology and practical rehabilitation strategies in ACL reconstruction. Overall, this review underscores the importance of an integrated approach that considers both the biological and mechanical aspects of ACL graft healing. Such an approach holds promise for advancing patient care and improving the success rates of ACL reconstruction surgeries.

Recent advancements in molecular diagnostics have highlighted the potential of circulating cell-free DNA (ccfDNA) as a biomarker in various diseases. This study aims to explore the role of ccfDNA in the early diagnosis, prognosis, and therapeutic monitoring of diseases, with a focus on oncology. We conducted a comprehensive review and analysis of current literature, combined with experimental approaches involving advanced genomic sequencing and bioinformatics techniques. Our research systematically characterizes the molecular composition of ccfDNA, including its origins, size distribution, and methylation patterns, across different disease states. Furthermore, we investigate the dynamics of ccfDNA release and clearance in the bloodstream, elucidating mechanisms that could influence its diagnostic accuracy. In the context of cancer, we analyze the correlation between tumor-derived ccfDNA and disease progression, response to therapy, and overall survival. Our findings indicate that ccfDNA holds promise as a non-invasive biomarker for early disease detection, real-time monitoring of treatment efficacy, and potentially, predicting relapse. This study contributes to the growing field of liquid biopsies and opens new avenues for personalized medicine, emphasizing the need for standardized protocols in ccfDNA analysis to maximize its clinical utility. The implications of our findings extend beyond oncology, suggesting potential applications of ccfDNA in other pathological conditions.

I present the first complete theory of ADHD. ADHD occurs when the organism adapts to try to maintain higher glucose levels than normal. The main mechanism via which this is done is by faster opening of KATP channels, mainly Kir6.2/SUR1, which increases the cell's intrinsic \xtblty. I show that this induces all of the symptoms of ADHD. The optimal treatment for ADHD already exists, since drugs of the sulfonylurea family close KATP channels and have been used for safely treating people for dozens of years.

S6K1 (ribosomal protein S6 kinase beta-1) is a critical serine/threonine kinase involved in the regulation of various cellular processes, including protein synthesis, gene expression, and metabolic homeostasis. The structural complexity of S6K1, with its multiple isoforms and unique kinase domain, allows for diverse functionality across different cellular contexts. In this review, we report the intricate structure of S6K1, highlighting the significance of its kinase domain. Additionally, we explored the tissue-specific expression and subcellular localization of S6K1 isoforms, emphasizing their distinct roles in cellular processes. The regulation of S6K1 extends beyond transcriptional control, with post-translational modifications such as phosphorylation and acetylation playing crucial roles in its activity and stability. The physiological implications of S6K1 are vast, influencing processes from cell growth and protein synthesis to immune responses and muscle hypertrophy. However, dysregulation of S6K1 is implicated in numerous pathological conditions, including cancer Studies reveal that altered S6K1 activity, particularly in its phosphorylation state, can lead to reduced neuronal viability, contributing to the progression of diseases such as Parkinson's. This manuscript synthesizes current knowledge on S6K1, offering insights into its multifaceted roles and highlighting its potential as a therapeutic target in various disease contexts.

Kiwifruit (Actinidia chinensis) cultivation is expanding worldwide being China, New Zealand and Italy the major producing countries. Halyomorpha halys, the brown marmorated stink bug, has been raising serious concerns to kiwifruit cultivation both in China and Italy. This study aimed at improving the chemical control efficacy against this pest by comparing two insecticide spray techniques (a conventional ray atomizer and a trumpet-modified atomizer adapted for localized spray application in kiwifruit). In fact, kiwifruit is often grown with a ‘pergola’ training system, which may reduce the effectiveness of insecticide penetration into the canopy. Experiments have been performed in naturally infested orchards of both Actinidia chinensis var. chinensis ‘Jintao’ and A. chinensis var. deliciosa ‘Hayward’ during two consecutive seasons. Furthermore, mesh cages, each containing three H. halys adults, were deployed in the orchards to assess the insects’ mortality at 1, 3, 7 and 10 days after the insecticide application with the two spray techniques. In the cultivar 'Jintao', the two systems performed similarly, while in the cultivar 'Hayward', an overall significantly higher insect mortality was recorded with the trumpet atomizer. The crop damage was also evaluated on both cultivars, simulating the grower insecticide applications with the two spray techniques. At harvest, no difference emerged between the spray techniques, which provided a significantly better protection compared to the untreated control. Further investigations in this direction are needed also considering the restriction of insecticidal active substances ongoing in European Union and the need to maximize the efficacy of the available tools.

Polar auxin transport (PAT) and auxin efflux carriers are known components controlling leaf complexity and venation patterns in some model plant species. Evidence indicates that PAT generates auxin converge points (CPs) that in turn lead to local leaf formation and internally into major vein formation. However, the role of PAT in more diverse leaf arrangements and vein patterns is largely unknown. We used pharmacological inhibition of PAT in developing pinnate tomato, trifoliate clover, palmate lupin, and bipinnate carrot leaves and observed dosage-dependent reduction to simple leaves in these eudicots. Leaf venation patterns changed from craspedodromous (clover, carrot), semi-craspedodromous (tomato) and brochidodromous (lupin) to more parallel patterning with PAT inhibition. The visualization of auxin responses in transgenic tomato plants showed that discrete and separate CPs in control plants were replaced by diffuse convergence areas near the margin. These effects indicate that PAT plays a universal role in the formation of different leaf and vein patterns in eudicot species via a mechanism that depends on the generation as well as separation of auxin CPs. Computer simulations indicate that variations in PAT can alter the number of CPs, corresponding leaf lobe formation and position of major leaf veins along the leaf margin in support of experimental results.

This study investigated the effect of partially replacing fish meal with krill and copepod meals in inert diets, co-fed with shrimp, on the growth and nutrient digestibility of long-snout seahorse (Hippocampus guttulatus). A control diet (Diet 1) using raw starch and four experimental diets with similar protein (≈44.8%) and energy (≈15.1 MJ/kg) levels were tested. Diet 2 used fish meal as the sole protein source, while in Diets 3-5, krill and copepod meals replaced 44% of the fish meal. Seahorses fed Shrimp + Diets 2-5 showed significantly higher growth rates (P < 0.05) than those fed Shrimp + Diet 1, though there were no significant growth differences among Diets 2-5. Digestibility of dry matter (46.1% to 72.2%), lipids (73.3% to 85.5%), crude protein (89.8% to 95.8%), energy (82% to 92.2%), and phosphorus (28.7% to 64.4%) varied with diet, being consistently lower in seahorses fed Shrimp + Diet 1. As an agastric species, H. guttulatus did not exhibit impaired digestibility for any of the tested nutrients, minerals, or energy. This study suggests that crustacean meals can effectively substitute fish meal in inert diets for this species, contributing to the sustainability and optimization of captive seahorse husbandry practices.

Deep brain stimulation of the subthalamic nucleus (STN-DBS) is a recognized treatment in Parkinson’s disease (PD), but knowledge is still limited regarding its possible impact on patients with behavioral addiction (BA) who undergo surgery. To assess this effect of STN-DBS, we prospectively studied the follow-up from 3-months pre-surgery to 12-months post-surgery of 12 PD patients with pre-existing BA (9 with pathological gambling disorder, 2 with hypersexual disorder, and 1 with compulsive shopping disorder) in a multicenter study. Patients were assessed for BA, for mental status with anxiety, depression, mania, emotional process and for PD, using the Unified Parkinson’s Disease Rating Scale motor examination (UPDRS3) and cumulative daily L-dopa equivalent daily dose (LEDD). Mean 6 and 12-month post-surgery scores were compared to baseline. After one year of STN-DBS, our results indicated that the treatment remained generally effective and stable. There was no effect on mania state or negative impact on motor signs. A significant improvement of the anxiety score was also observed. The STN is an interesting target for new therapeutic stimulation in BA and works to investigate the precise target in this neurobiological architecture should be performed in the future.

Soluble inorganic pyrophosphatase (sPPase), a pyrophosphate hydrolase, has been shown to play an important role in several physiological processes such as plant growth and development, metabolic activities, and response to abiotic stresses, but sPPase has been less studied in woody plants. In order to preliminarily verify whether the soluble inorganic pyrophosphatase in Eucommia ulmoides Oliver (E. ulmoides) has a role in resistance to drought stress, we amplified the EuSIP5 (E. ulmoides soluble inorganic pyrophosphatase 5) cDNA sequence from E. ulmoides by RT-PCR. In order to study the drought resistance of the Eucommia ulmoides soluble inorganic pyrophosphatase 5 (EuSIP5) gene, the cDNA sequence of the EuSIP5 gene was amplified from Eucommia ulmoides Oliver by RT-PCR. Bioinformatic analysis indicated that EuSIP5 might be an unstable amphipathic protein, mainly located in the cytoplasm. E. ulmoides, the highest expression of the EuSIP5 gene was found in male leaves and pericarp from April to October, and in leaves of E. ulmoides in July and September. During drought, the expression of EuSIP5 in seedling E. ulmoides leaves was significantly higher than that of the control. An overexpression vector harboring the EuSIP5 was constructed and introduced into the tobacco (Nicotiana tabacum cv. Xanthi). We found that during drought stress, the appearance of wilting was delayed by 4 days in transgenic tobacco plants (OE) compared with wild type plants (WT), and the expression of drought-related genes DET2, CYP85A1, P5CS, ERF1 and F-box was found to be increased in transgenic tobacco leaves, as well as the enzyme activities of the protective enzymes: peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) were significantly higher than those of the WT, and the malondialdehyde (MDA) content was lower than the WT. These results indicated that the introduction of EuSIP5 gene into tobacco increased the expression of drought-related genes, enhanced the anti-membrane oxidation ability of tobacco, and reduced the oxidative stress damage, thus improving the drought resistance of tobacco.

Wheat is used for making many food products due to its diverse quality profile found among different wheat classes. Since laboratory analysis of these end-use quality traits is costly and time-consuming, genetic dissection of the traits is preferential. This study used a genome-wide association study (GWAS) of 10 end-use quality traits, including kernel protein, flour protein, flour yield, softness equivalence, solvent’s retention capacity, cookie diameter, and top-grain, in soft red winter wheat (SRWW) adapted to US southeast. The GWAS study included 266 SRWW genotypes that were evaluated in two locations over two years (2020-2022). A total of 27,466 single nucleotide markers were used and a total of 80 significant marker-trait associations were identified. There were 13 major effect quantitative trait loci (QTLs) explaining > 10% phenotypic variance out of which 12 were considered as novel. Five of the major effect QTLs were found to be stably expressed across multiple datasets, and four showed associations with multiple traits. Candidate genes were identified for eight of the major-effect QTLs including genes associated with starch biosynthesis and nutritional homeostasis in plants. These findings increase genetic comprehension of these end-use quality traits and could potentially be used for improving the quality of SRWW.

The mosquito-borne zoonotic nematode parasites, Dirofilaria immitis and Dirofilaria repens, primarily affect dogs. In recent years, their distribution has expanded due to various factors influencing vector-borne pathogens. This study aimed to investigate the comparative prevalence of infection in dogs and humans within a hyperenzootic region of Europe and to estimate the proportional relationship between infection prevalence in dogs and humans, in the concept of “One Health”. To this end, 604 blood samples from dogs and 625 serum samples from humans living in the Thrace region of northeastern Greece were collected. Dog samples were examined using Kott’s test for Dirofilaria spp. microfilariae and a commercial serological test for D. immitis antigen. Human sera were analysed by western blot for both parasites. The overall prevalence of infection in dogs was 177 (29.3%), with 173 (28.6%) testing positive for D. immitis and 7 (1.3%) for D. repens, including 6 (1%) cases of mixed infection. Specific IgG antibodies were detected in 42 (6.7%) human samples, with 24 (3.8%) positive for D. immitis and 18 (2.9%) for D. repens. The infection proportion in humans was 23.4% of the corresponding canine infection, indicating a high risk of human infection in this hyperenzootic region.

This article examines the biological characteristics and potential medicinal uses of the endemic plant species Scorzonera uzbekistanica, found only in Uzbekistan. The plant contains significant amounts of bio active compounds such as inulin, flavonoid, and oxyacetylene in its roots. These compounds are known for their antioxidant, anti-inflammatory, and prediction properties. Traditionally used to alleviate digestive issues, this plant's potential in modern medicine should be further supported by scientific research. The study calls upon the scientific community to explore the potential applications of this endemic plant.

The neural system has limited resources to interact with the environment, and this is where attention comes into play. Attention is a complex process that provides numerous benefits, but there has been a longstanding debate about the mechanisms involved in attentional orientation. Two widely proposed mechanisms are top-down and bottom-up, which relate to the organism's current behavioral goal and the salience of environmental stimuli, respectively. However, distinguishing between these mechanisms can be challenging in both behavioral and neural studies. This review introduces a new conceptualization in which both top-down and bottom-up mechanisms are viewed as aspects of the same process. This unified approach aims to prepare the organism to face stimuli and events based on its prior experiences, immediate (the current task that he/she is aware of), and innate (to be alive) demands.

Butirosins are naturally occurring aminoglycoside (AG) antibiotics featuring a 4,5-disubstituted 2-deoxystreptamine (2-DOS) with a (2S)-4-amino-2-hydroxybutyrate (AHBA) side chain. This side chain has been shown to confer resistance against AG modifying enzymes, leading to ongoing studies on the butirosin biosynthetic pathway and the corresponding enzymes. Butirosin is produced by Niallia (formerly Bacillus) circulans and Bacillus vitellinus, with most research focused on the former strain. To date, no whole genome analysis has been performed on B. vitellinus. In this study, we sequenced the complete genome of B. vitellinus NBRC 13296 and performed a comparative analysis of different butirosin biosyntheric gene clusters (BGCs), including those from N. circulans. The complete genome of B. vitellinus NBRC 13296 comprises a 6,331,192 base circlular chromosome with a GC content of 52.68%. The annotation revealed the presence of 5,605 CDSs, 70 tRNA genes, 30 rRNA genes, and 3 ncRNA genes in NBRC 13296. The highest dDDH and ANI values between NBRC 13296 and the most closely related type strain, Paenibacillus chitinolyticus KCCM 41400, were 97.8% and 98.66%, respectively. Based on these genome-based comparative analyses, we propose reclassifying B. vitellinus NBRC 13296 as P. chitinolyticus. Genome mining revealed 18 gene clusters encoding the biosynthesis of diverse secondary metabolites in the genome of B. vitellinus NBRC 13296, indicating the enormous biosynthetic potential of this strain. The predicted structural diversity of secondary metabolites includes aminoglycoside, PKS, NRPS, PKS-NRPS hybrids, metallophores, phosphonates, terpenes, β-lactones, and RiPP peptides. We then comparatively characterized butorosin BGCs previously studied in several N. circulans strains. Additionally, comparative genome analysis revealed complete butirosin BGCs identified from P. chitinolyticus KCCM 41400, P. chitinolyticus NRRL B-23119, P. chitinolyticus NRRL B-23120, P. chitinolyticus B-14908, P. chitinolyticus YSY-3.1, P. chitinolyticus JMW06, Paenibacillus sp. GbtcB18, Paenibacillus sp. HGH0039, Paenibacillus sp. MZ04-78.2. Finally, we identified the core region consisting of BtrS, BtrN, BtrM, BtrL, BtrA, BtrB, BtrC, BtrD, BtrD, BtrE, BtrF, BtrG, BtrH, BtrI, BtrI, BtrJ, BtrK, BtrO, BtrP, and BtrV, followed by an upstream region organizing BtrQ, BtrW, BtrX, BtrY, and BtrZ in the same transcriptional direction and sequential genetic arrangement, and a downstream region organizing various proteins based on BtrT, BtrR2, BtrU, and BtrR1. Our study provided insights into the reclassification of B. vitellinus NBRC 13296 to P. chitinolyticus and suggested the need for continued studies on butirosin biosynthesis from an enzymatic perspective.

Consumer perception of beef is heavily influenced by overall meat quality, a critical factor in the cattle industry. Genomics has the potential to improve important beef quality traits and identify genetic markers and causal variants associated with these traits by genomic selection (GS), and genome-wide association studies (GWAS) approaches. Transcriptomics, proteomics, and metabolomics provide insights into underlying genetic mechanisms by identifying differentially expressed genes, proteins, and metabolic pathways linked to quality traits, complementing GWAS data. Leveraging these functional genomics techniques can optimize beef cattle breeding for enhanced quality traits to meet high-quality beef demand. This paper provides a comprehensive overview of the current state applications of the omics technologies in uncovering functional variants underlying beef quality complexities. By highlighting the latest findings from GWAS, GS, transcriptomics, proteomics, and metabolomics studies, this work seeks to serve as a valuable resource fostering a deeper understanding of the complex relationships between genetics, gene expression, protein dynamics, and metabolic pathways in shaping beef quality.

Mitochondria are dynamic organelles that can adjust and respond to different stimuli within a cell. This plastic ability allows them to effectively coordinate several cellular functions in cells and becomes particularly relevant in highly complex cells such as neurons. An imbalance in mitochondrial dynamics can disrupt mitochondrial function, leading to abnormal cellular function and ultimately to a range of diseases, including neurodegenerative disorders. Regulation of mRNA transport and local translation inside neurons is crucial for maintaining the proteome of distal mitochondria, which is vital for energy production and synaptic function. A significant portion of the axonal transcriptome is dedicated to mRNAs for mitochondrial proteins, emphasizing the importance of local translation in sustaining mitochondrial function in areas far from the cell body. In neurons, local translation and the regulation of mRNAs encoding mitochondrial-shaping proteins could be essential for synaptic plasticity and neuronal health. The dynamics of these mRNAs, including their transport and local translation, may influence the morphology and function of mitochondria, thereby affecting the overall energy status and responsiveness of synapses. Comprehending the mitochondria-related mRNAs regulation and local translation, as well as its influence on mitochondrial morphology near the synapses will help to better understand neuronal physiology and neurological diseases where mitochondrial dysfunction and impaired synaptic plasticity play a central role.

Autoimmune Glomerulonephritis represents a homogeneous area of renal pathology with clinical relevance for either the numerical impact or difficulties in treatment. SLE and Lupus nephritis are the most frequent occurrences. They are characterized by glomerular deposition of circulating autoantibodies that recognize glomerular antigens Technologies for studying tissue and circulating antibodies have evolved over the years and culminated with the direct analysis of the antigen-antibody complex in renal bioptic fragments. Mass-spectrometry of immunoprecipitates from the renal tissue is, in fact, the most recent evolution that eliminates the need for tissue microdissection. The results obtained with this technique have led to the discovery of several new glomerular antigens in Membranous Nephropathy and in Lupus Nephritis, two conditions whose pathogenesis remained unknown for decades, Peptide and protein arrays (Immunopeptidomics and immunoproteomics, respectively) for the characterization of circulating autoantibodies represent the new frontier for identifying autoantibodies. Immunopeptidomics consists in 7.5 million aligned peptides of 16 amino acids each which cover the whole human proteome; immunoproteomics utilizes a chip containing structured proteins, 26.000 overall. Both are very sensitive techniques that allow the characterization of hundreds of new antibodies in circulation. The contribution of arrays to amplify the panel of potential autoantibodies involved in glomerulonephritis is expected to be relevant for improving our knowledge of the pathogenesis of several autoimmune conditions.

Autism spectrum disorder (ASD) involves social communication difficulties and repetitive behaviors, with a growing prevalence worldwide. Symptoms include cognitive impairments, gastrointestinal (GI) issues, feeding difficulties, and psychological problems. A significant concern in ASD is food selectivity, leading to nutrient deficiencies. Common GI issues in ASD, such as constipation and irritable bowel syndrome, stem from abnormal gut flora and immune system dysregulation. Sensory sensitivities and behavioral challenges exacerbate these problems, correlating with neurological symptom severity. Children with ASD also exhibit higher oxidative stress due to low antioxidant levels like glutathione. Therapeutic diets, including ketogenic, high-antioxidant, gluten-free, and casein-free, show potential in managing ASD symptoms like behavior, communication, GI issues, and oxidative stress, though the evidence is limited. Various studies have focused on different populations, but there is increasing concern about the impact on children. This review examines the effect of food selectivity, GI problems, and antioxidant deficiencies in ASD and revises the effectiveness of various therapeutic diets, highlighting the need for further research to confirm their long-term benefits.

Abstract: Founders of quantum theory have pointed out the irreducibility of living systems to quantum mechanics. Quantum mechanics is never violated they rightfully argue but works under a set of active macroscopic constraints that selectively order the behavior of the micro components and these constraints are not reducible to the reversible, causally closed formalism of quantum mechanics. The whole of biological evolution is characterized by the production of increasingly more highly-ordered levels of such macroscopic constraints and biological evolutionary theory has no account for this either. It is assumed outside its theoretical first principles. In fact this generative macroscopic ordering, the production of increasingly more highly differentiated levels of order—more things, more kinds of things, cannot be explained within biological theory because it is generic to the universe itself. Irreducible to quantum mechanics this begs the question of fundamentality in physics. Fundamental laws in a universe defined by active macroscopic ordering, level-building, and differentiation must be able to causally cross levels, and more importantly explain why there is any level-building at all to begin with. The laws of thermodynamics are fundamental laws in this sense, and 4th Law path selection explains the generative universal ordering and differentiation (“complexification”).

The main goal of this study was to pinpoint functional candidate genes associated with multiple economically important traits in Nellore cattle. After quality control, 1 830 genomic regions sourced from 52 scientific peer-reviewed publications were used in this study. From these, a total of 8 569 positional candidate genes were annotated for reproduction, 11 195 for carcass, 5 239 for growth, and 3 483 for morphological traits, and used in the overrepresentation analysis. The significant genes (adjusted p-values<0.05) identified in the overrepresentation analysis underwent prioritization analyses, and an enrichment analysis of the prioritized overrepresented candidate genes was performed. The prioritized candidate genes were: GFRA4, RFWD3, SERTAD2, KIZ, REM2, and ANKRD34B for reproduction, RFWD3, TMEM120A, MIEF2, FOXRED2, DUSP29, CARHSP1, OBI1, JOSD1, NOP58, and LOXL1-AS1 for carcass, ANKRD34B, and JOSD1 for growth traits, and no gene prioritized for morphological traits. The functional analysis pinpointed the following genes: KIZ (plays crucial role in spindle organization, essential in forming a robust mitotic centrosome), DUSP29 (involved in muscle cell differentiation), and JOSD1 (involved in protein deubiquitination, thereby improving growth). The enrichment of the functional candidate genes identified in this study highlights that these genes play an important role in the expression of reproduction, carcass, and growth traits in Nellore cattle.

This experiment aimed to determine the temperature limits beyond which seahorse growth and reproduction become suboptimal due to climate change. Four temperatures (16, 20, 24, and 28°C) were tested to evaluate their effect on juvenile (1-56 days post-parturition (DPP)) and adult (one year old) seahorses. Additionally, the reproductive performance of adults was observed. Another experiment measured oxygen consumption (MO2) in the same age groups and temperatures. Adults showed significantly higher growth rates at 20 and 24°C compared to 16 and 28°C. Adult mortality rates were 0%, 0%, 6.2%, and 62.5% at the respective temperatures. Juvenile growth performance was higher at 20°C and 24°C but significantly lower at 16°C, with survival rates of 8%, 62%, 10%, and 0%, respectively. Oxygen consumption increased with age and temperature, ranging from 106.3±3.1 to 203.3±3.1 μmol O2/g BW/h at 16°C, and from 128.8±3.8 to 273.3±3.1 μmol O2/g BW/h at 28°C. The study highlights that juvenile and adult H. guttulatus have narrow thermal boundaries, beyond which reproduction, growth, and survival are seriously affected. Under climate change, the species appears unable to cope, potentially leading to their rapid disappearance.

This study evaluated the presence of Human Papillomavirus (HPV) and Bovine Leukemia Virus (BLV) DNA in breast cancer (BC) samples from women in Northeastern Brazil. We analyzed 186 formalin-fixed, paraffin-embedded tissue block samples using PCR and qPCR to detect HPV and BLV DNA. BLV DNA was not detected in any BC patients. However, HPV DNA was found in 46.2\% of the BC samples: 44.6\% were positive for HPV16, 3.5\% for HPV31, and 2\% were coinfected with HPV16 and HPV31. Additionally, we found an association between HPV-positive BC and Ki-67 expression (P=0.03, OR=1.85; CI=1.02-3.34), as well as between HPV31-positive BC and mitotic index (P=0.047). Although BLV was not detected in BC patients from Northeastern Brazil, the association between HPV and cell proliferation markers warrants further investigation, as HPV may contribute to the aggressiveness of BC in these patients.

Inflammatory bowel diseases (IBD) involve chronic inflammation of the gastrointestinal tract, where effector CD4+ T-cells play a central role. Thereby, the recruitment of T-cells into the colonic mucosa represents a key process in IBD. We recently found that CCR9 and DRD5 might form a heteromeric complex on the T-cell surface. The increase of CCL25 production and the reduction in dopamine levels associated with colonic inflammation represent a dual signal stimulating the CCR9:DRD5 heteromer, which promotes the recruitment of CD4+ T-cells into the colonic lamina propria. Here, we aimed to analysing the molecular requirements involved in the heteromer assembly as well as to determining the underlying cellular mechanisms involved in the colonic tropism given by the stimulation of the CCR9:DRD5 complex. The results show that dual stimulation of the CCR9:DRD5 heteromer potentiates the phosphorylation of the myosin light chain 2 (MLC2) and the migration speed in confined microchannels. Accordingly, disrupting the CCR9:DRD5 assembly induced a sharp reduction in the pMLC2 in vitro, decreased the migratory speed in confined microchannels and dampened the recruitment of CD4+ T-cells into the inflamed colonic mucosa. Furthermore, in silico analysis confirmed that the interface of interaction of CCR9:DRD5 is formed by the transmembrane segments 5 and 6 from each protomer. Our findings demonstrated that the CCR9:DRD5 heteromeric complex plays a fundamental role in the migration of CD4+ T-cells into the colonic mucosa upon inflammation. Thereby, disassembling the formation of the CCR9:DRD5 represents an important therapeutic opportunity to treat IBD.

To evaluate dryland wheat genotypes' performance to different pre-crop and residue management under dryland conditions, a split-split plot experiment based on RCB design with three replications was conducted for two years (2017-2018 and 2018-2019). The site of study's long-term average precipitation, temperature, and relative humidity are 376 mm, 9 ◦C, and 50%, respectively. Wheat-wheat and vetch-wheat cropping systems were considered in the main plots, different wheat and vetch residue levels including 0, 2, and 4 t ha-1 were located in subplots, and five dryland wheat genotypes including Sadra, Hashtroud, Baran, Varan, and Ohadi were allocated in sub-sub plots. Results indicated that leaf chlorophyll content index (CCI) and stomatal conductance (gs) were greater in the vetch-wheat cropping system compared to the wheat monoculture system for all genotypes. Normalized differences vegetative index (NDVI) of genotypes improved by applying the crop residue. Over two years, the application of crop residues resulted in higher variable fluorescence at J and I steps as well as an increase in the photosynthesis performance index (PI). Varan and Baran genotypes stood out as the superior genotype, exhibiting the highest values physiological characteristics and grain yield under the application of 4 t ha-1 of vetch residue. Grain filling rate (GFR) was reduced, while grain filling duration (GFD) was increased with increasing the crop residue levels. The enhanced grain yield of wheat genotypes grown under vetch residue attributed to the factors such as improvement in leave pigments and photosynthetic efficiency which facilitate longer grain filling duration with high grain weight. As a result, it is advisable to adopt a vetch-wheat cropping system with a high proportion of crop residue in dryland regions to achieve increased and sustainable wheat production.

Cardiovascular disease is a leading cause of morbidity and mortality. New research elucidates in-creasingly complex relationships between cardiac and metabolic health, giving rise to new possible therapeutic targets. Sphingolipids are a heterogenous class of bioactive lipids with critical roles in normal human physiology. They have also been shown to play both protective and deleterious roles in the pathogenesis of cardiovascular disease. This review will explore the roles of Sphingolipids in vascular, cardiac, and metabolic pathologies and will evaluate the therapeutic potential in targeting Sphingolipids with the aim of prevention and reversal of cardiovascular disease, in order to improve long-term cardiovascular outcomes.

Determining the estrous cycle stages in mice is essential for optimizing breeding strategies, synchronizing experimental time lines, and facilitating studies in behavior, drug testing, and genetics. It is critical for reducing the production of genetically unmodified offspring in the generation and investigation of genetically modified animal models. An accurate detection of the estrus cycle is particularly relevant in the context of the 3Rs—Replacement, Reduction and Refinement. The estrous cycle, encompassing the reproductive phases of mice, is key to refining experimental designs and addressing ethical issues related to the use of animals in research. This study presents results from two independent laboratories on the efficacy of the Mouse Estrus Detector (MED) from Elmi Ltd. (Latvia) for the accurate determination of the estrus phase. The female mice of five strains/stocks (CD1, FVB/N, C57Bl6/J, B6D2F1, and Swiss) were used. The results showed that the MEDProTM is a low -traumatic, simple, rapid and painless method of estrus detection that supports the principles of the 3Rs. The use of the MEDProTM for estrus detection in mice caused minimal stress, enhanced mating efficiency, facilitated an increase in the number of embryos for the in vitro fertilization and allowed the production of the desired number of foster animals.

Understanding the structural intricacies of RNA molecules is essential for deciphering numerous biological processes. Traditionally, scientists have relied on experimental methods to gain insights and draw conclusions. However, the recent advent of advanced computational techniques has significantly accelerated and refined the accuracy of research results in several areas. A particularly challenging aspect of RNA analysis is the prediction of its secondary structure, which is crucial for elucidating its functional role in biological systems. This paper deals with the prediction of pseudoknots in RNA, focusing on two types of pseudoknots: K-type and M-type pseudoknots. Pseudoknots are complex RNA formations in which nucleotides in a loop form base pairs with nucleotides outside the loop and thus contribute to essential biological functions. Accurate prediction of these structures is crucial for understanding RNA dynamics and interactions. Building on our previous work in which we developed a framework for the recognition of H- and L-type pseudoknots, an extended grammar-based framework tailored to the prediction of K- and M-type pseudoknots is proposed. This approach uses syntactic pattern recognition techniques and provides a systematic method to identify and characterize these complex RNA structures. Our framework uses context-free grammars (CFGs) to model RNA sequences and predict the occurrence of pseudoknots. By formulating specific grammatical rules for type K and type M pseudoknots, we enable efficient parsing of RNA sequences to recognize potential pseudoknot configurations. This method ensures an exhaustive exploration of possible pseudoknot structures within a reasonable time frame. In addition, the proposed method incorporates essential concepts of biology, such as base pairing optimization and free energy reduction, to improve the accuracy of pseudoknot prediction. These principles are crucial to ensure that the predicted structures are biologically plausible. By embedding these principles into our grammar-based framework, we aim to predict RNA conformations that are both theoretically sound and biologically relevant.

Triple-negative breast cancer (TNBC) lacks the expression of estrogen receptors, human epidermal growth factor receptor 2, and progesterone receptors (PR). TNBC has the poorest prognosis among breast cancer subtypes and is more likely to respond to immunotherapy due to its higher expression of PD-L1 and a greater percentage of tumor-infiltrating lymphocytes. Immunotherapy has revolutionized TNBC treatment, especially with the FDA's approval of pembrolizumab (Keytruda) combined with chemotherapy for advanced cases, opening new avenues for treating this deadly disease. Although, immunotherapy can significantly improve patient outcomes in a subset of patients, achieving the desired response rate for all remains an unmet clinical goal. Strategies that improve responses to immune checkpoint blockade, including combining immunotherapy with chemotherapy, molecularly targeted therapy, or radiotherapy may improve response rates and clinical outcomes.  In this review, we provide a short background on TNBC and immunotherapy and explore the different types of immunotherapy strategies that are currently being evaluated in TNBC. Additionally, we review why combination strategies may be beneficial, provide an overview of the combination strategies, and discuss the novel immunotherapeutic opportunities that may be approved in the near future for TNBC.

Economic, social, and environmental sustainability are the results of efforts aiming to improve all aspects of milk production, respecting animal welfare and improving herd health. An epidemi-ological study was designed to assess the role of contagious pathogens (S. aureus and S.agalactiae) in a cohort of 120 dairy herds located in the southern regions of Italy. Milk quality was assessed using certified methods, and the prevalence of mastitis pathogens in bulk tank milk was determined using quantitative polymerase chain reaction. Welfare scores were assessed using a scoring card that have more than 100 items, including animal-based measurements. Statistical analyses were performed using general lineal model and logistic regression procedures. The results showed that S. aureus had a significant negative effect on the amount of milk components delivered to the dairy plant, and on the level of welfare. Whereas, the presence of S.agalactiae did not show any significant association. The major risk factors associated with the presence of S.aureus were also identified to help prioritise control programs. These results support the “One Sustainability” approach im-plying that an increase of animal productivity is related to the improvement of animal health and welfare and potentially leading to the mitigation of greenhouse gas emissions.Economic, social, and environmental sustainability are the results of efforts aiming to improve all aspects of milk production, respecting animal welfare and improving herd health. An epidemi-ological study was designed to assess the role of contagious pathogens (S. aureus and S.agalactiae) in a cohort of 120 dairy herds located in the southern regions of Italy. Milk quality was assessed using certified methods, and the prevalence of mastitis pathogens in bulk tank milk was determined using quantitative polymerase chain reaction. Welfare scores were assessed using a scoring card that have more than 100 items, including animal-based measurements. Statistical analyses were performed using general lineal model and logistic regression procedures. The results showed that S. aureus had a significant negative effect on the amount of milk components delivered to the dairy plant, and on the level of welfare. Whereas, the presence of S.agalactiae did not show any significant association. The major risk factors associated with the presence of S.aureus were also identified to help prioritise control programs. These results support the “One Sustainability” approach im-plying that an increase of animal productivity is related to the improvement of animal health and welfare and potentially leading to the mitigation of greenhouse gas emissions.

Growth monitoring of crops is a crucial aspect of precision agriculture, essential for optimal yield prediction and resource allocation. Traditional crop growth monitoring methods are labor-intensive and prone to errors. This study introduces an automated segmentation pipeline utilizing multi-date aerial images and ortho-mosaics to monitor the growth of cauliflower crops (Brassica Oleracea var. Botrytis) using an object-based image analysis approach. The methodology employs YOLOv8, Grounding Detection Transformer with Improved Denoising Anchor Boxes (DINO), and the Segment Anything Model (SAM) for automatic annotation and segmentation. The YOLOv8 model was trained using aerial image datasets, which then facilitated the training of the Grounded Segment Anything Model framework. This approach generated automatic annotations and segmentation masks, classifying crop rows for temporal monitoring and growth estimation. The study’s findings utilized a multi-modal monitoring approach to highlight the efficiency of this automated system in providing accurate crop growth analysis, promoting informed decision-making in crop management and sustainable agricultural practices. Results indicate consistent and comparable growth patterns between aerial images and ortho-mosaics, with significant periods of rapid expansion and minor fluctuations over time. The results also indicated a correlation between the both the time and method of observation which paves a future possibility of integration of such techniques aimed at increasing the accuracy in crop growth monitoring based on automatically derived temporal crop row segmentation masks.

Vaginitis is a widespread issue for women worldwide, yet current diagnostic tools are lacking. Bacterial vaginosis (BV) is the most prevalent type of vaginitis, found in 10-50% of reproductive aged women. Current diagnostic methods for BV rely on clinical criteria, microscopy, or detection of a few microbes by qPCR. However, many vaginal infections lack a single etiological agent and are characterized by changes in the vaginal microbiome community structure (e.g. BV is defined as a loss of protective lactobacilli resulting in an overgrowth of anaerobic bacteria). Shotgun metagenomic sequencing provides a comprehensive view of all the organisms present in the vaginal microbiome (VMB) allowing for a better understanding of all potential etiologies. Here, we describe a robust VMB metagenomics sequencing test with a sensitivity of 93.1%, specificity of 90%, a negative predictive value of 93.4%, and a positive predictive value of 89.6% certified by Clinical Laboratory Improvement Amendments (CLIA), the College of American Pathologist (CAP) and the Clinical Laboratory Evaluation Program (CLEP). We sequenced over 7,000 human vaginal samples with this pipeline and described general findings and comparison to US census data.

Global climatic pressures and increased human demands create a modern necessity for efficient and affordable plant phenotyping unencumbered by arduous technical requirements. The analysis and archival of imagery have become easier as modern camera technology and computers are leveraged. This facilitates the detection of vegetation status and changes over time. Using a custom lightbox, an inexpensive camera, and common software, turfgrass pots were photographed in a greenhouse environment over an 8-week experiment period. Subsequent imagery was analyzed for area of cover, color metrics, and sensitivity to image corrections. Findings were compared to active spectral reflectance data and previously reported measurements of visual quality, productivity, and water use. Results indicate that Red Green Blue-based (RGB) imagery with simple controls is sufficient to measure the effects of plant treatments. Notable correlations were observed for corrected imagery, including between a percent yellow color area classification segment (%Y) with human visual quality ratings (VQ) (R = -0.89), the dark green color index (DGCI) with clipping productivity in mg d-1 (mg) (R = 0.61), and an index combination term (COMB2) with water use in mm d-1 (mm) (R = -0.60). The calculation of green cover area (%G) correlated with Normalized Difference Vegetation Index (NDVI) (R = 0.91) and its RED reflectance spectra (R = -0.87). A CIELAB b*/a* chromatic ratio (BA) correlated with Normalized Difference Red-Edge index (NDRE) (R = 0.90), and its Red-Edge (RE) (R = -0.74) reflectance spectra, while a new calculation termed HSVi correlated strongest to the Near-Infrared (NIR) (R = 0.90) reflectance spectra. Additionally, COMB2 significantly differentiated between the treatment effects of date, mowing height, deficit irrigation, and their interactions (p < 0.001). Sensitivity and statistical analysis of typical image file formats and corrections that included JPEG (JPG), TIFF (TIF), geometric lens correction (LC), and color correction (CC) were conducted. Results underscore the need for further research to support image corrections standardization and better connect image data to biological processes. This study demonstrates the potential of consumer-grade photography to capture plant phenotypic traits.

This study analyzed methodologies for identifying β-casein genotypes in dairy cows and the purity of A2 fermented milk. The research was justified by the need for quick and accurate techniques for herd screening and A2 milk quality control. The methodology included the development of an ELISA test to detect β-casein A1 in fermented milk and the evaluation of a commercial Lateral Flow Immuno Assay (LFIA) to identify A2A2 genotypes and the purity of A2 milk. The results showed that the ELISA test had high sensitivity and specificity (100%) in detecting β-casein A1. The LFIA test was 100% effective in identifying A2A2 genotypes and verifying the purity of A2 milk with a minimum detectable contamination of 5% for raw milk and 10% for fermented milk. Compared to traditional genotyping, these methods proved to be highly reliable and more practical for large-scale screening. It was concluded that the ELISA and LFIA were valuable tools for ensuring the quality and authenticity of A2 milk, meeting the demands of producers and consumers for safe and healthy dairy products.

Toxoplasma gondii is a protozoan and the etiologic agent of toxoplasmosis causes high mortality in immunocompromised individuals and newborns. Despite the medical importance of toxoplasmosis, few drugs are available for its treatment, which are associated with side events and parasite resistance. Here, we show a screening of molecules present in Covid-Box as a way of discovering new hits with anti-T. gondii activity. Covid-Box contains 160 molecules with known or predicted activity against SARS-CoV-2. Our analysis selected 23 Covid-Box molecules that can inhibit the tachyzoite forms of the RH strain of T. gondii in vitro by more than 70% at 1 µM after seven days of treatment. The inhibitory curves showed that most of these molecules inhibited the proliferation of tachyzoites with IC50 values below 0.80 µM; Cycloheximide and (-)-Anisomycin were the most active drugs, with IC50 values of 0.02 μM. Cell viability assays showed that the compounds are not toxic at active concentrations, and most are highly selective for parasites. Overall, all 23 compounds were selective, and for three of them (Apilimod, Midostaurin, and Salinomycin), this is the first report of activity against T. gondii. To better understand the effect of the drugs, we analyzed the effect of six of them on the ultrastructure of T. gondii using transmission electron microscopy. The main changes observed in parasite morphology after treatment with the selected drugs were changes in cell division and parasite organelles.

Steroidogenic factor 1 (SF-1) is a nuclear receptor that regulates steroidogenesis and reproductive development. NR5A1/SF-1 variants are associated with a broad spectrum of phenotypes across individuals with disorders of sex development (DSD). Oligogenic inheritance has been suggested as an explanation. However, testing the impact of specific gene variants involved in a network remains difficult. To confirm the hypothesis that NR5A1/SF-1-related DSD follow an oligogenic mode of inheritance, we investigated a constellation of gene variants identified in a 46,XY severely undervirilized individual. Candidate genes were revealed by whole exome sequencing, and pathogenicity was predicted by different in silico tools. We found variants in NR1H2 and INHA associated with steroidogenesis, sex development, and reproduction. Functional testing was conducted in cell models. Novel SF-1 and NR1H2 binding sites in the AR and INHA gene promoters were found. Transactivation studies showed that wild-type NR5A1/SF-1 regulates INHA and AR gene expression, while the NR5A1/SF-1 variant had decreased transcriptional ac-tivity. NR1H2 was found to regulate AR gene transcription; however, the NR1H2 variant showed normal activity. This study expands the NR5A1/SF-1 network of interacting partners while strengthening the hypothesis that the broad phenotype observed in 46,XY DSD individuals may be caused by oligogenic pathogenicity.

The blood that remains in the placenta and umbilical cord after the birth of the infant is referred to as umbilical cord blood (UCB), which has the advantages of convenient source and less ethical controversy. MicroRNA (miRNA) is a class of non-coding RNAs that can regulate gene expression after translation and transcription. Alterations in miRNA expression involve changes in a range of gene expression pathways. An increasing body of research indicates that miRNAs are implicated in many human disorders and are involved in almost all biological medium processes. Due to their high stability in UCB, circulating miRNAs are considered promising biomarkers for diagnosis and disease prognosis. Herein, we conduct a comprehensive review to summarize the roles of diverse UCB miRNAs from different perspectives, including UCB miRNA as biomarker for developmental projections and disease diagnosis, the relationship between maternal gestational status and neonatal UCB miRNA expression, the therapeutic function of exosomal UCB miRNA, miRNA expression profiles in UCB cell lineages. Taken together, miRNAs in UCB are very promising biomarkers for indicating physiological states, disease states, drug responses, development states, or health conditions, which can help in diagnosing, classifying, and predicting diseases, prognosis, or providing personalized treatment plans for patients.

Pro-inflammatory cytokines play a role in the failure of β-cells in type 1 and type 2 diabetes. While existing data from ‘omics’ experiments allow for some understanding of the molecular mechanisms behind cytokine-induced dysfunction in β-cells, no report thus far has provided information on the direct imaging of the β-cell landscape with nanoscale resolution following cytokine exposure. In this study, we use Airyscan-based optical super-resolution microscopy of Insulinoma 1E (INS-1E) cells to investigate the structural properties of two subcellular membranous compartments involved in the production, maturation and secretion of insulin-containing granules, the Endoplasmic Reticulum (ER) and the Golgi Apparatus (GA). Our findings reveal that exposure of INS-1E cells to IL-1β and IFN-γ for 24 hours leads to significant structural alterations of both compartments. In more detail, both ER and GA fragment and give rise to vesicle-like structures with markedly reduced characteristic area and perimeter and increased circularity with respect to the original structures. These findings complement the molecular data collected thus far on these compartments and their role in β-cell dysfunction using and lay the ground for future optical-microscopy-based ex-vivo and in-vivo investigations.

Abstract: This study aimed to assess the agronomic performance and yield stability of elite white yam (Dioscorea rotundata) genotypes across diverse Nigerian environments. Twenty-five genotypes were evaluated at three locations in two consecutive growing seasons, 2022 and 2023. The genotypes were assessed for their agronomic performance, tuber yield and related traits using various analytical tools such as AMMI (Additive main effects and multiplicative interaction) and MTSI (Multi-trait stability index). The AMMI analysis revealed significant differences among the genotypes and across the environ-ments for all traits (p

: This study focused on assessing whether the inclusion of probiotics (B. subtilis) as feed additives during the preweaning stage can enhance the body weight and metabolic condition of neonatal calves. A total of 50 Holstein calves, all born on the same farm, were randomly divided into two homogeneous treatment groups after birth. The probiotic was administered 24 hours post-birth, signifying the start of the experimental period. The Control Group (CG), consisting of 25 calves, was fed with a milk replacer, while the Treatment Group (TG), also with 25 calves, received milk replacer supplemented with 7.5 mL/calf/day of B. subtilis probiotic, in accordance with the manufacturer's guidelines. It took one month to collect the animals. Body weight was measured at birth for all animals. A local veterinarian, working on the farm, conducted daily health checks of the calves, recording health parameters and any antibiotic treatments. Blood samples were collected from each calf at birth and again after 30 days by puncturing the jugular vein, using 10 mL evacuated serum tubes before morning feeding. Significant differences in body weight were observed at 30 days of age (p < 0.05). The CG's average body weight (BW) was 52.71 kg (±10.68), compared to 58.49 kg (±14.21) in the TG, a difference of 5.78 kg or an increase of 9.88%. Additionally, the TG exhibited significantly lower aspartate transaminase (AST) activity at 30 days of age (p < 0.001), with an average of 51.02 IU/L (±5.16) compared to 72.00 IU/L (±12.68) in the CG, a 41.12% decrease. The TG also had a significantly higher average phosphorus (P) concentration (p < 0.001) of 3.27 mmol/L (±0.22), a 9.36% increase over the CG's 2.99 mmol/L (±0.27). A significant increase in gamma-glutamyl transferase activity was observed in the TG compared to the CG (p < 0.001), with a 64.48% (or 26.29 IU/L) increase. The TG's average GGT activity was 40.64 IU/L (±22.43), while in the CG it was 14.35 IU/L (±3.45). Additionally, there was a significant 17.7% decrease (or 10.02 g/L) in total protein (TP) concentration in the TG compared to the CG, with the TG averaging 46.32 g/L (±17.43) and the CG 56.34 g/L (±9.81). Overall, dairy calves who were given conventional milk replacer along with a daily dose of 7.5 mL of B. subtilis probiotic experienced enhanced growth performance and a more favorable metabolic profile. Their body weight exhibited a 9.88% growth, but the activity of AST decreased by 41.12%, suggesting a decreased likelihood of liver damage. Additionally, their protein digestibility was 17.7% greater compared to the control group. In addition, B. subtilis had a positive impact on the overall concentration of phosphorus.

The vaccines have to undergo independent testing at National Control laboratory (NCL) for various quality control tests to ensure safety, purity, and potency before market authorization and surveillance testing. In this study, five different inactivated cell culture rabies vaccine preparations (Aluminum adjuvanted and non-adjuvanted) marketed in India were evaluated using compendial safety test of mouse inoculation test (MIT) for residual live virus. Investigations were done to see the interference of aluminum adjuvant in rabies vaccine preparation to ensure safety as per the in-vivo compendial test for virus inactivation. After intracerebral injections, mice received the aluminum phosphate adjuvanted inactivated rabies vaccine (ARV) had neurological complications that resulted in their deaths between days 4 and 10. Further investigations were carried out to find out the root cause for mice death including fluorescent antibody test (FAT), rabies virus amplification test. Additionally, alamar blue based cell cytoxicity assay was carried out using BHK-21 cells to estimate the cell toxicity of aluminum phosphate adjuvanted inactivated rabies vaccine. The present study highlighted the challenges in ensuring the safety of adjuvated inactivated rabies vaccine at level of NCL. Further, study demonstrated the toxicity of aluminum under both in vitro and in vivo settings, sparking controversy regarding the safety of aluminum adjuvants in vaccines at the suggested compendial limit of 1.25 mg/single human dosage.

Various strategies have been employed to improve the reliability of 2D, 3D, and co-culture in vitro models of nonalcoholic fatty liver disease, including using extracellular matrix proteins, such as collagen I, to promote cell adhesion. While studies have demonstrated the significant benefits of culturing cells on collagen I, its effects on HepG2 and HepaRG cell lines after exposure to palmitate (PA) have not been investigated. Therefore, this study aimed to assess the effects of PA-induced lipotoxicity in HepG2 and HepaRG cultured in the absence or presence of collagen I. HepG2 and HepaRG cultured in the absence or presence of collagen I were exposed to PA, followed by analyses that assessed cell proliferation, viability, adhesion, cell death, mitochondrial respiration, reactive oxygen species production, gene and protein expressions, and triacylglycerol accumulation. Culturing HepG2 on collagen I was associated with increased cell proliferation, adhesion, expression of integrin receptors, and improved cellular spreading compared to culturing them in the absence of collagen I. PA-induced lipotoxicity was greater in collagen I-cultured HepG2 than those cultured in the absence of collagen I. In summary, the present study demonstrated for the first time that collagen I-cultured HepG2 exhibited exacerbated cell death following exposure to PA through integrin-mediated death. The findings from this study may serve as a caution to those using 2D models or 3D scaffold-based models of HepG2 in the presence of collagen I.

A multi-environment evaluation of sorghum genotypes was conducted across six environments in the 2021 main growing season in a randomized complete block design with three replications. The objectives of the study were to estimate the magnitude of genotypes by environment interaction (GEI) and grain yield stability of drought-tolerant sorghum genotypes across different environments. Data were subjected to analysis of variance, Additive Main Effects and Multiplicative Interaction (AMMI), and GGE biplot analysis. Combined analysis of variance revealed significant variations among genotypes, environments, and GEI for yield and yield-related traits, indicating that these factors significantly affected grain yield. The maximum mean grain yield value of genotypes due to the mean effect of the environment was obtained from G1 (5119.93kg ha-1), followed by G14 (4834.57 kg ha-1), and G18 (4801.20 ha-1), while the least mean grain yield was obtained from G3 (3314.50 kg ha-1). The multiplicative variance of the treatment sum of squares due to GEI was partitioned into four principal component axes (PCA). Sum squares of the first and second interaction principal component axis (IPCA) explained 71.07% and 17.50% of the GEI variation, respectively. The IPCA1&2 mean squares were highly significant (P≤0.01), indicating the adequacy of the AMMI model with the first two IPCAs for cross-validation of grain yield variation. The magnitude of the GEI sum squares was 3.9 times that of the genotype sum squares for grain yield, indicating the presence of substantial differences in genotypic responses across environments. The results of cultivar superiority measure (Pi), yield stability index (YSI), AMMI stability value (ASV), regression coefficient (bi), and deviation from regression (S2di) depicted that genotypes G18, G22, G31, and 32 were the most stable genotypes for grain yield and biomass yield, respectively. AMMI2 biplot showed Jinka, Alduba, and Kako were the most discriminating environments as indicated by the long distance from the origin; whereas testing locations Meioso and Gato with short vector length indicated that these locations had less discriminating power on the genotypes' performance. The study has provided precious information on the yield stability status of the sorghum genotypes and the best environments for future improvement programs in Ethiopia.

Sclerotinia minor Jagger is a phytopathogenic fungus that causes lettuce drop, a serious problem of lettuce (Lactuca sativa L.) production. The control of this pathogen is challenging because of the presence of resistance of sclerotia, which can survive in the soil under favourable conditions. In Bulgaria, the management of lettuce drop relies primarily on the strategic application of synthetic fungicides. To find alternative methods for disease management, four bacterial isolates were screened for antagonism against S. minor. This study reports the in vitro evaluation of the antifungal activity of Bacillus subtilis, Priestia megaterium, Bacillus safensis, and Bacillus mojavensis against S. minor. The molecular identification of the isolates involved in the activity was examined by 16s rRNA sequencing. Isolated bacterial strains produced indole-3-acetic acid (IAA) in a medium supplemented with 0.1% L-tryptophan. The ability of those strains to increase the mobility of phosphorus and zinc has been elucidated. The production of siderophores has been confirmed on CAS (Chrom azurol S) medium. The inhibitory action showed by the filtrates of the bacterial growth broths against S. minor and points out the nature of the molecules involved. The evaluation of the antifungal activity was carried out in vitro and pot experiments. This study determined the effect of growth-promoting rhizobacteria on the development of lettuce. This research focuses mainly on the development of biocontrol strategies for the management of lettuce drop in greenhouses.

Tamil Nadu, a state in southern India, is bereft of any native milch or dual-purpose cattle breed and the state depends chiefly on crossbred cattle for milk production. As most (90 percent) of the milk production in the state was contributed by Jersey crossbred cattle, the genetic evaluation study was intended to assess the milk production traits viz., test-day milk yields (TDMYs), total milk yield (TMY) and 305-day milk yield (305MY) in these crossbred cattle reared under farmers’ rearing environment. Data on monthly TDMYs (TDMY1 to TDMY10) of Jersey crossbred cows (n=81,653) reared by the farmers over 24 years (1999-2022) were collected for evaluation. The influence of non-genetic factors viz., location (agroclimatic zones), period and season of calving, and parity on TDMYs, TMY, and 305MY were assessed, and the variance and covariance components for genetic parameters (heritability and genetic correlation) of the traits were estimated by the method of restricted maximum likelihood (REML) under multivariate animal model and by random regression model (RRM). The overall means of various TDMYs ranged from 4.98 (TDMY10) to 9.95 kg (TDMY2) and, for TMY and 305MY the means were 2480.33 and 2393.71 kg respectively. The heritabilities estimated by animal model for TDMYs were high and ranged from 0.39 ± 0.01 to 0.47 ± 0.01 and were fairly less variable throughout the lactation while the estimates obtained by RRM were medium to high and ranged between 0.28 ± 0.01 and 0.65 ± 0.02 for different days in milk (DIMs). The heritability estimated for TMY and 305MY were 0.48 ± 0.01 and 0.49 ± 0.01 respectively. The estimates of heritability in mid-lactation yields were comparatively little less than those at the beginning and the end of lactation. The high estimates of heritabilities for TDMYs, TMY, and 305MY obtained in the present study offered an ample scope for improving milk production through selective breeding. Genetic correlations between TDMYs estimated by animal model were positive and moderate to high, with a range between 0.47 ± 0.01 and 0.99 ± 0.00. Considerably higher estimates (0.97-0.99) were observed between adjacent TDMYs showing strong genetic associations. By RRM, genetic correlations estimated between DIMs were positive, except for a few estimates and the magnitude of genetic correlation decreased with an increase in the interval between the DIMs. The high genetic correlation observed between the TDMYs in early and mid stages of lactation with TMY and 305MY suggested that these test day yields could be used favorably for the early selection of cows for milk production which facilitates reduction in the generation interval and consequently increase the annual genetic gain for the milk production traits.

Xyleborus beetles, a diverse group of ambrosia beetles, present significant challenges to forestry and agriculture due to their damaging burrowing behavior and symbiotic relationships with fungi. This review synthesizes current knowledge on the biology, ecology, and management of Xyleborus. We explore the beetles' life cycle, reproductive strategies, habitat preferences, and feeding habits, emphasizing their ecological and economic impacts. Control and management strategies, including preventive measures, chemical and biological control, and integrated pest management (IPM), are critically evaluated. Recent advances in molecular genetics and behavioral studies offer insights into genetic diversity, population structure, and host selection mechanisms. Despite progress, managing Xyleborus effectively remains challenging. This review identifies future research needs and highlights innovative control methods, such as biopesticides and pheromone-based trapping systems.

Trehalose, a disaccharide made of two D-glucose molecules, is found in various organisms like bacteria, yeast, fungi, insects, and plants. It serves as an energy and carbon source in plants and has signaling functions in yeast and plants, affecting metabolic pathways and growth regulation. It also plays a role in protecting proteins and cell membranes from damage caused by stress. This study aims to determine the best level of trehalose supplementation in layer diets between 34 to 49 weeks of age, due to limited literature on its impact on laying hen productivity. Experimental diets were based on nutritional recommendations and included six trehalose levels. The study was conducted over five periods of 21 days each, evaluating various parameters such as feed intake, egg production, egg quality, and bone characteristics. The results showed that trehalose supplementation influenced the performance of laying hens, as well as some aspects of egg and bone quality.

To investigate the effects of predation of the paddle crab Charybdis japonica on the culture and survival of scallops (Mizuhopecten yessoensis) during bottom culture, we investigated the behavioral characteristics of three sizes (small, medium, and large) of scallops in response to exposure to crabs. We also measured the activities of superoxide dismutase, catalase, arginine kinase, and octopine dehydrogenase in the gill, adductor muscle, and mantle of scallops before and after exposure to predation. Tissues that showed significant differences between control and test specimens were selected for deep sequencing of the transcriptome to identify and validate the key genes that were sensitive to predation. We found when M. yessoensis is stimulated by the presence of predators, its behavioral characteristics and related physiological and ecological indexes undergo significant changes. The results are relevant for developing specifications for M. yessoensis seedling casting during bottom culture.

Fungi inhabiting deep subseafloor sediments have been shown to possess anaerobic methane (CH₄) production capabilities under atmospheric conditions. However, their ability to produce CH₄ under in situ conditions with high hydrostatic pressure (HHP) remains unclear. Here, Schizophyllum commune 20R-7-F01, isolated from ~ 2 km below the seafloor, was cultured in Seawater Medium (SM) in culture bottles fitted with sterile syringes for pressure equilibration. Subsequently, these culture bottles were transferred into 1-liter stainless steel pressure vessels at 30 °C for 5 days to simulate in situ HHP and anaerobic environments. Our comprehensive analysis of bioactivity, biomass, transcriptomics revealed that S. commune not only survived but significantly enhanced CH₄ production, reaching approximately 2.5 times higher levels under 35 MPa HHP compared to 0.1 MPa standard atmospheric pressure. Pathways associated with carbohydrate metabolism, methylation, hydrolase activity, cysteine and methionine metabolism, and oxidoreductase activity were notably activated under HHP. Specifically, key genes involved in fungal anaerobic CH₄ synthesis, including methyltransferase mct1 and dehalogenase dh3, were upregulated 7.9- and 12.5-fold, respectively, under HHP. Enhanced CH₄ production under HHP was primarily attributed to oxidative stress induced by pressure, supported by intracellular Reactive Oxygen Species (ROS) levels and comparative treatments with cadmium chloride and hydrogen peroxide. These findings suggest that fungi persisting in deep-sea sediments over millions of years have the potential to significantly contribute to global CH₄ flux.

This study analyzed methodologies for identifying β-casein genotypes in dairy cows and the purity of A2 fermented milk. The research was justified by the need for quick and accurate techniques for herd screening and A2 milk quality control. The methodology included the development of an ELISA test to detect β-casein A1 in fermented milk and the evaluation of a commercial Lateral Flow Immuno Assay (LFIA) to identify A2A2 genotypes and the purity of A2 milk. The results showed that the ELISA test had high sensitivity and specificity (100%) in detecting β-casein A1. The LFIA test was 100% effective in identifying A2A2 genotypes and verifying the purity of A2 milk with a minimum detectable contamination of 5% for raw milk and 10% for fermented milk. Compared to traditional genotyping, these methods proved to be highly reliable and more practical for large-scale screening. It was concluded that the ELISA and LFIA were valuable tools for ensuring the quality and authenticity of A2 milk, meeting the demands of producers and consumers for safe and healthy dairy products.

Fagao is one of China's traditional gluten free staple foods made by rice or corn flour. Corn Fagao prepared by co-fermentation with sourdough and yeast exhibits better quality and less staling compared to traditional yeast-fermented Fagao. The physicochemical properties of corn Fagao batter during the sourdough-yeast co-fermentation were investigated. The results showed that compared with yeast fermentation, the gas production and viscosity of the batter increased by co-fermentation. The co-fermented batter shows higher hydrolysis of starch and less amylose content. The integrity of starch granules in co-fermented batter was damaged more seriously, the crystallinity and short-range ordered structure were less than yeast fermented batter, even though the crystal structure type of starch did not change obviously. The peak viscosity, minimum viscosity, final viscosity, decay value and recovery value of corn batter are reduced by co-fermentation, which can improve the thermal stability of the batter and slow down the aging. Co-fermentation also results in a more pronounced reduction in protein subunit contents than yeast fermentation. The changes in the physicochemical properties of corn Fagao batter help explain the quality improvement of corn Fagao made from co-fermentation method, and may provide theoretical references for co-fermentation with sourdough and yeast to other gluten free foods.

The high transmissibility, rapid evolution, and immune escape of SARS-CoV-2 variants can influence the course of infection and, in turn, morbidity and mortality in COVID-19, posing a challenge in controlling transmission rates and contributing to the emergence and spread of new variants. The factors that shape viral genetic variation are consequently essential for understanding the evolution and transmission of SARS-CoV-2, especially in vaccinated individuals where immune response plays a role in the progression and spread of this disease. This study assessed. In this context, we evaluated the impact of immunity induced by the CoronaVac vaccine (Butantan/Sinovac) on intra-host genetic diversity, analyzing 118 whole-genome sequences of SARS-CoV-2 from unvaccinated and vaccinated patients infected with the Gamma variant. Vaccination with CoronaVac favors negative selection at the intra-host level in different genomic regions and prevent greater genetic diversity of SARS-CoV-2, which may help reduce the emergence of new mutations, reinforcing the importance of vaccination in reducing virus transmission.

Comprehending the replication kinetics of SARS-CoV-2 variants helps explain why certain var-iants spread more easily and are more contagious, and pose significant health menaces to the global populations. The replication kinetics of Malaysian isolates of the Alpha, Beta, Delta, and Omicron variants were studied in the Vero E6 cell line. Their replication kinetics were deter-mined using the plaque assay, quantitative real-time PCR (qRT-PCR), and viral growth curve. The Beta variant exhibited the highest replication rate at 24 hours post-infection (h.p.i), as evi-denced by the highest viral titers and lowest viral RNA multiplication threshold. The plaque phenotypes also varied among the variants, in which the Beta and Omicron variants formed the largest and smallest plaques, respectively. All studied variants showed strong cytopathic effects after 48 h.p.i. The whole-genome sequencing highlighted cell-culture adaptation, where the Beta, Delta and Omicron variants acquired mutations at the multibasic cleavage site after three cycles of passaging. The findings suggest a strong link between the replication rates and their respec-tive transmissibility and pathogenicity. This is essential in predicting the impacts of the upcom-ing variants on the local and global populations and is useful in designing preventive measures to curb the virus outbreaks.

One of the main objectives of developing new anti-cancer vaccine strategies is to effectively induce CD8+ T cell-mediated anti-tumor immunity. Live recombinant vectors, notably Listeria mono-cytogenes, have been shown to elicit a robust in vivo CD8+ T cell response in preclinical settings. Significantly, it has been demonstrated that Listeria induces inflammatory/immunogenic cell death mechanisms such as pyroptosis and necroptosis in immune cells that favorably control immunological responses. Therefore, we postulated that the host's response to Listeria-based vec-tors and the subsequent induction of CD8+ T cell-mediated immunity would be compromised by the lack of regulatory or effector molecules involved in pyroptosis or necroptosis. To test our hy-pothesis, we used recombinant L. monocytogenes carrying the ovalbumin gene (LM.OVA) to vac-cinate wild-type (WT), caspase-1/11-/-, gsdmd-/-, ripk3-/-, and mlkl-/- C57Bl/6 mice. We performed an in vivo cytotoxicity assay to assess the efficacy of OVA-specific CD8+ T lymphocytes in eliminat-ing target cells in wild-type and genetically deficient backgrounds. Furthermore, we evaluated the specific anti-tumor immune response in mice inoculated with B16F0 and B16F0.OVA mela-noma cell lines. Our findings demonstrated that while caspase-1/11 and GSDMD deficiencies in-terfere with the rapid control LM.OVA infection, of the KO seems to contribute to the early acti-vation of OVA-specific CTL responses. In contrast, individual deficiency of each one of these pro-teins negatively impact the generation of long-lasting effector CD8+ T cells.

This review provides a comprehensive analysis of the pelvic plexus and its regulation across various mammalian species, including rats, cats, dogs, and pigs. The pelvic and hypogastric nerves play crucial roles in regulating pelvic functions such as micturition, defecation, and erection. The anatomical organization of these nerves varies, forming either well-defined ganglia or complex plexuses. Despite these variations, the neurons within these structures are consistently regulated by key neurotransmitters—norepinephrine and acetylcholine. These neurons also possess receptors for testosterone and prolactin, particularly in rats, indicating the significant role of these hormones in neuronal function and development. Moreover, neuropeptides such as vasoactive intestinal peptide (VIP), substance P, neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL), and calcitonin gene-related peptide (CGRP) are co-released with neurotransmitters to modulate pelvic functions. This review highlights the complex interplay between neurotransmitters, neuropeptides, and hormones in regulating pelvic physiology and emphasizes the importance of hormonal regulation in maintaining the functionality and health of the pelvic plexus across different species.

Antibiotic prescription is the first line of treating bacterial diseases. But it affects the composition and function of non-targeted intestinal microbiota. Antimicrobial resistance will be the cause for the death of millions of people worldwide. By considering collateral damage of antibiotics, searching an alternative antimicrobial agent will be a solution to minimize the problems. Probiotics, prebiotics and synbiotics are an alternative to antibiotics. Thanks to GIT microbes as they are responsible for metabolism, energy production, protection from pathogens and immunomodulation. So, it is appreciable if the gut microbiota modulated through consumption of probiotics, prebiotics and synbiotics. Probiotic effects are genus and species specific. To get good health benefit of probiotics, safety of probiotics should be assessed before using them. This is mandatory for prebiotics and synbiotic selection. So this review paper gives information about side effects of antibiotics on gut microbiota, description of probiotics, prebiotics and synbiotics as alternative to antibiotics in modulating GIT microbiota, techniques of assessing safety of probiotics and health benefits of probiotics, prebiotics and synbiotics with scientific evidence in clinical trial and some disagreement and challenges of using probiotics, prebiotics and synbiotics.

(1) Background: Triggering receptor expressed on myeloid cells 2 (TREM2) is involved in neu-roinflammation and HIV-associated neurocognitive impairment (NCI). People with HIV (PWH) using cannabis exhibit lower inflammation and neurological disorders. We hypothesized TREM2 dysfunction mediates HIV neuropathogenesis and can be reversed by cannabinoids. (2) Methods: EcoHIV-infected wildtype (WT) and TREM2R47H mutant mice were used to study HIV's impact on TREM2 and behavior. TREM2 and related gene expressions were examined in mono-cyte-derived macrophages (MDMs) from PWH (n=42) and people without HIV (PWoH; n=19) with varying cannabis use via RNA sequencing and qPCR. Differences in membrane-bound and soluble TREM2 (sTREM2) were evaluated using immunocytochemistry (ICC) and ELISA. (3) Results: EcoHIV increased immature and C-terminal fragment forms of TREM2 in WT mice but not in TREM2R47H mice, with increased IBA1 protein in TREM2R47H hippocampi, correlating with worse memory test performance. TREM2 mRNA levels increased with age in PWoH but not PWH. Cannabidiol (CBD) treatment increased TREM2 mRNA alone and with IL1β. RNA-seq showed upregulation of TREM2-related transcripts in cannabis-using PWH compared to naïve controls. IL1β increased sTREM2 and reduced membrane-bound TREM2, effects partially re-versed by CBD. (4) Conclusions: These findings suggest HIV affects TREM2 expression modu-lated by cannabis and CBD, offering insights for therapeutic strategies.

Long COVID is a phenomenon in which individuals experience persistent symptoms after recovering from COVID-19. The symptoms are discovered to be unique for every individual and can affect multiple organs systems in the body. This study aims to identify effective natural remedies for long COVID by analyzing hub genes associated with the symptoms of the condition and evaluating the repurposed drugs catered and used for treating the symptoms of long COVID. The most common and prevalent symptoms of long COVID were identified; Fatigue, Shortness of Breath, Loss of Smell, Headache, Brain Fog, Chest Pain, Insomnia, Heart Palpitations, Dizziness, Joint Pain, Depression, Anxiety, Tinnitus, Loss of Appetite. Hub genes for each of the symptoms provided an insight on the key biological pathways of the symptom. Repurposed drugs identified, provided the template to identify the natural compounds with similar structure as a potential therapeutic drug. The natural compounds were retrieved using fingerprint search of the repurposed drugs from the NPASS Database. The findings of this study suggest several natural remedies for each symptom based on the molecular docking of the hub gene and natural compound using iGEMDOCK. The identified natural remedies may hold promise in treating long COVID, but further research is required to explore the efficacy and effectiveness of the proposed natural compounds. The results of the study pose important implications for the development of effective treatments for long COVID.

This literature review presents a comparative analysis of cranial morphology in Allosaurus, focusing on its implications for feeding mechanics. Allosaurus, a large theropod dinosaur from the Late Jurassic period, exhibits distinct cranial features that are pivotal for understanding its predatory behaviors and ecological role. By examining fossilized skulls and employing advanced imaging techniques, this research identifies key morphological traits such as tooth structure, jaw articulation, and cranial robusticity. The analysis reveals that Allosaurus had a specialized cranial architecture that supported a powerful bite force and efficient processing of prey. Comparative metrics are applied to assess similarities and differences with other theropods, such as Tyrannosaurus rex, to elucidate variations in feeding strategies among theropods. The findings suggest that Allosaurus possessed adaptations that allowed it to exploit a diverse range of prey, indicating a versatile feeding strategy. These results enhance our understanding of theropod evolution and ecological interactions, providing insights into the functional significance of cranial morphology in dinosaurian predation. This review underscores the importance of cranial features in reconstructing the behavior and diet of extinct species and contributes to the broader field of vertebrate functional morphology.

Oxidative stress and chronic inflammation create a perpetual cycle in the elderly, where impaired immune function amplifies susceptibility to oxidative damage, and oxidative stress further weakens the immune response. This cycle is particularly detrimental to the respiratory system of the elderly, which is an easy target for constant exogenous harmful attacks during cold/flu season or under heavy air pollution. Herbal medicines that protect respiratory function are seen as safer alternatives to conventional therapies; however, there is limited availability of scientifically validated, safe, and effective natural supplements for these conditions. In this study, we evaluated a standardized bioflavonoid composition, UP446, which contains bioactives from the roots of Scutellaria baicalensis and the heartwoods of Acacia catechu, as a natural and nutritional supplement for its antioxidative and immunoregulatory effects in oxidative stress-accelerated aging and chemically induced immune suppression mouse models. Immunosenescence was induced by continual subcutaneous injection of D-galactose (D-Gal) at a dose of 500 mg/kg/day in CD-1 mice. UP446 was administered orally at doses of 100 mg/kg and 200 mg/kg starting in the fifth week of immunosenescence induction. The study lasted a total of ten weeks. All mice received a quadrivalent influenza vaccine 14 days before the end of the study. Whole blood, serum, spleen homogenate, and thymus tissues were used for analysis.Cyclophosphamide (Cy)-induced immunosuppression was triggered by three consecutive injections of cyclophosphamide at 80 mg/kg/day, followed by oral administration of UP446 for 18 days at doses of 100 mg/kg and 200 mg/kg. Blood was collected from each animal at necropsy, and serum was isolated for IgA and IgG ELISA analysis. UP446 was found to improve immune response, as evidenced by stimulation of innate (NK cells) and adaptive immune responses (T cells and cytotoxic T cells), an increase in antioxidant capacity (glutathione peroxidase), preservation of vital immune organs (the thymus), and a reduction in NFκB. UP446 also increased serum levels of IgA and IgG. The findings presented in this report demonstrate the antioxidative, anti-inflammatory, and immune-regulatory activities of UP446, suggesting its potential use in respiratory conditions involving immune stress due to aging, oxidative stress, and/or pathogenic challenges.

This study employed High-Performance Liquid Chromatography coupled with Diode Array De-tector and Mass Spectrometry (HPLC-DAD-MS) to characterize bioactive compounds in olive leaf extracts. The analysis revealed a diverse metabolic profile, predominantly composed of tyrosol, flavonoids, and hydroxycinnamic acid. The antioxidant activity of the extracts demonstrated their efficacy, with variations observed between neutral and acidic pH conditions. Encapsulation was explored as a preservation method, successfully maintaining the integrity of phenolic compounds in both liquid and powdered forms. All four extracts have an undeniable antimicrobial effect. Moreover, the antibacterial activity tested on E. coli ATCC 25922 and S. aureus ATCC 25923 strains significantly improved post-encapsulation, particularly for the acidic pH extract. The findings suggest that encapsulation can potentially conserve and/or enhance the antimicrobial activity of olive leaf extract, paving the way for developing functional products with improved shelf life and nutritional properties. Further investigations are recommended to elucidate underlying mecha-nisms and optimize encapsulation processes to maximize the benefits of olive leaf extract in di-verse applications.

Wastewater genomic surveillance is an efficient and economic approach to track infectious diseases and can help detect unusual epidemiological events. The current study uses this approach for the first time in Morocco to monitor the epidemiology of SARS-CoV-2 and Influenza A, B and RSV virus infections during the third wave of COVID-19 caused by the Omicron variant. The virome was concentrated from wastewater collected from two sewer sheds of two cities, Agadir and Inezgane, using the PEG/NaCl method. The viral copy number was determined using qRT-PCR. All 26 samples in both cities were tested positive for SARS-CoV-2 at various viral loads. For Influenza A virus, two samples were tested positive in Inezgane. The estimated SARS-CoV-2 viral RNA copy numbers observed were then used to estimate the number of infected individuals using the SEIR model. The number of viral copies of SARS-CoV-2 for Agadir and Inezgane varied from 82932 ± 15479 to 507865 ± 58759 copies/L and 61244 ± 8552 to 497506 ± 18061 copies/L, respectively. The estimated number of cases correlates positively with the number of reported cases. Next Generation Sequencing showed that samples contain the following two variants: BA.1 and BA.2 that have been detected in clinical samples. This study demonstrates the effectiveness and feasibility of wastewater genomic surveillance in monitoring pathogens such as SARS-CoV-2 in Morocco.

Drug residues that contaminate food and water represent a serious concern for human health, since their irrational use is constantly increasing the amplitude of exposure. Multiple sources contribute to the overall exposure to contaminants, including agriculture, domestic use, personal, public, and veterinary healthcare, increasing the possible origin of contamination. In this review, we focus on crop pesticides and veterinary drug residues because of their extensive use in modern agriculture and farming, which ensures food production and security for the ever-growing population around the world. We discuss crop pesticides and veterinary drug residues with respect to their worldwide distribution and impacts, with special attention on their harmful effects on human reproduction and embryo development, as well as their link to epigenetic alterations, leading to intergenerational and transgenerational diseases. Among the contaminants, the most commonly implicated in causing such disorders are organophosphates, glyphosate and antibiotics, with tetracyclines being the most frequently reported. This review highlights the importance of finding new management strategies for pesticides and veterinary drugs. Moreover, due to the still limited knowledge on inter- and transgenerational effects of these contaminants, we underlie the need to strengthen research in this field, so as to better clarify the specific effects of each contaminant and their long-term impact.

Comparative analysis of mortality rates between public and private healthcare systems remains a critical yet complex area of study. This article synthesizes findings from extensive academic literature and empirical studies to examine the relationship between healthcare system type and patient outcomes across various countries. By focusing on mortality rates as a key indicator, we elucidate the nuanced impacts of public versus private healthcare delivery. Our analysis reveals the multifaceted challenges inherent in both systems and their implications for patient survival. We conclude by identifying crucial gaps in current research and proposing policy recommendations to optimize healthcare outcomes in diverse global contexts.This revised abstract:Starts with a strong opening statement that immediately captures the topic's importance. Clarifies the article's methodology (synthesizing existing literature and studies).Specifies the focus on mortality rates as the primary outcome measure.Hints at the complexity of the findings, avoiding oversimplification.Ends with a clear statement about the article's contributions to both research and policy.

Background: Mules are used worldwide for agrarian purposes such as plowing and harvesting crops. In this study, mules were used for plowing and cultivation. While mules are believed to be less susceptible to diseases and fatigue than horses, no scientific investigation has been conducted to support this belief. The objective was to assess effect of work-associated fatigue on hematological and biochemical parameters in working mules in Mexico. Methodology: Six healthy mules 6.5 to 23 years, 3.5 BCS, were studied. Three blood samples/day were taken at the beginning and end of the work season. One sample taken at the beginning of the day, middle, and end (3 hours apart). Biochemical parameters, including the concentrations of albumin, globulin, total protein, glucose, alkaline phosphatase (ALP), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), total bilirubin, lactate, creatinine kinase (CK), urea, creatinine, calcium, sodium, potassium, total carbon dioxide (tCO2), and al-bumin-to-globulin ratio, were determined using the veterinary Skyla-VB1 automated analyzer (Lite-On Technology Corporation, Hsinchu-Taiwan), and blood lactate concentration using the handheld Lactate Scout Vet. Results: Significant increases in concentrations AST (P < 0.001), BUN (P < 0.001), calcium (P = 0.002), creatine phosphokinase CPK U/L (P < 0.001), tCO2 (P = 0.006), and urea (P < 0.01), were observed at the end of the work season. However, sodium, potassium, and chloride were significantly decreased at the end of the work season. No treatment or time-by-treatment interaction effect was observed for ALP, creatinine, globulin, or the albu-min-to-globulin ratio. Conclusions: The significant decrease in mineral and electrolytes (Na, K, and Cl) at the end of work season could suggest change in electrolytic balance during the strenuous plowing season. A higher concentration of tCO2 at end of the season is a good indicator of metabolic acid-base disturbances. Results indicate use of mules for plowing fields disrupts the mules' micromineral and electrolyte profiles. Increased CPK levels demonstrate that the workload was excessive. This study furthers our understanding of biochemical responses in mules.

Cyclocarya paliurus tea, also known as "sweet tea", a health substitute tea with Cyclocarya paliurus leaves as raw material, is famous for its unique nutritional benefits and flavor. However, due to the unique "bittersweet" of Cyclocarya paliurus tea, it is still unable to fully satisfy consumers' high-quality taste experience and oral pleasure. The taste and sensory quality of Cyclocarya paliurus tea are mainly determined by secondary metabolites. Therefore, this study aimed to explore the metabolites in Cyclocarya paliurus leaves during their growth process, particularly the composition and variation of sweet and bitter taste compounds, by combining multi-platform metabolomics analysis with an electronic tongue system and molecular docking simulation technology. The results indicated that there were significant differences in the contents of total phenols, flavonoids, polysaccharides and saponins of C. paliurus leaves in different growing months, among which the bioactive components in leaves harvested in July were the highest, and the best balance between sweetness and bitterness was achieved. A total of 575 secondary metabolites were identified as potential active metabolites related to sweet/bitter taste using nontargeted metabolomics based on UHPLC-MS/MS analysis. Molecular docking technology was utilized to study the interactions between candidate metabolites and the sweet receptor T1R2/T1R3 and the bitter receptor T2R4/T2R14. Six key compounds with high sweetness and low bitterness were successfully identified by using computational simulation analysis, including cis-Anethole, Gluconic acid, beta-D-Sedoheptulose, Asparagine, Proline, and Citrulline, which may serve as candidates for taste modification in Cyclocarya paliurus leaves. These findings provide a new perspective for understanding the sweet and bitter taste characteristics metabolites that contribute to the distinctive sensory quality of Cyclocarya paliurus leaves.

The aim of this study was to investigate and effect of application of different conditions of freeze-drying on the kinetics of process and the sorption properties of dried apples. Slices of apple were frozen and subjected to freezing-drying process with different combination of a shelf temperature (-20, 10, 20, 30°C) and pressure (37, 63, 103, 165 Pa). During the freeze-drying, the temperature in the centre of material was recorded. The moisture content in dried material and changes of water content dried apples stored at humidity of 75.3% were obtained. The Midilli et al. model was used to describe drying kinetics of the freeze-drying with good fit. Drying time decreased from 660 (variant with a constant shelf temperature of 30°C, pressure 63 Pa) to 1305 minutes (variant with temperatures −20: 10: 20: 30°C, pressure 63 Pa). For this reason, the most favourable experimental parameters were: temperature of 30°C and pressure of 63 Pa. However, application of these parameters caused higher absorption of water vapour during storage. Therefore, the selection of freeze-drying parameters should also be related to the expected properties of the final product.

Expansins are cell wall (CW) proteins that mediate the CW loosening and regulate salt tolerance in a positive or negative way. The transcription of Populus trichocarpa expansin A6 (PtEXPA6) was downregulated upon prolonged duration of salt stress (48 h) after a transient increase induced by NaCl (100 mM). The role of PtEXPA6 in salt tolerance was determined by transferring the PtEXPA6 gene into the hybrid species Populus alba × P. tremula var. glandulosa clone 84K (84K) and Populus tremula × P. alba INRA '717-1B4' (717-1B4). The PtEXPA6-transgenic poplars of 84K and 717-1B4 showed a greater reduction in photosynthesis, stem height and diameter growth under salt stress and a reduced capacity for ROS scavenging by antioxidant enzymes, which is associated with excessive Na+ accumulation in roots and shoots. NMT flux recordings showed that the radial translocation of Na+ salt into the root xylem and real-time Na+ translocation from roots to leaves were greater in PtEXPA6-transgenic poplars than in wild-type poplars. Analysis of comparative contractility and comparative extensibility of intact root tips indicated that PtEXPA6 increased the CW loosening in the transgenic poplars of 84K and 717-1B4. Noteworthy, the PtEXPA6-promoted CW loosening was shown to facilitate radial and longitudinal transport of Na+ in transgenic poplars. We conclude that overexpression of PtEXPA6 leads to CW loosening that facilitates radial translocation of Na+ into the root xylem and subsequent Na+ translocation from roots to leaves, resulting in excessive Na+ accumulation and consequently reduced salt tolerance in transgenic poplars.

Neurologic disorders such as traumatic brain injury, multiple sclerosis, Alzheimer’s disease, and drug-resistant epilepsy have a high socioeconomic impact around the world. Current therapies for these disorders are often not effective. This creates a demand for the development of new therapeutic approaches to treat these disorders. Recent data suggest that autoreactive naturally occurring immunoglobulins produced by subsets of B cells called B1 B cells combined with complement are actively involved in the processes of restoration of neuronal functions during pathological conditions and remyelination. The focus of this review is to discuss the possibility of creating specific therapeutic antibodies that can activate and fix complement to enhance neuronal survival and promote central nervous system repair after injuries associated with many types of neurodegenerative diseases.

Long COVID is the collective term used to describe the persistence of symptoms in individuals who have recovered from SARS-CoV-2 infection. Some people who have healed from COVID-19 continue to experience new or persistent symptoms that last for weeks or months. While Long COVID is becoming more widely recognized, its fundamental causes and effective therapies remain largely unclear. This study aims to explore the potential of phytochemical medications as a cutting-edge therapeutic strategy for treating Long COVID, also known as post-acute sequelae of SARS-CoV-2 infection (PASC).G protein-coupled receptors (GPCRs), which play a critical role in cell signaling, were the focus of this study. GPCRs are known to be involved in immunological and inflammatory responses and are considered potential targets for Long COVID therapy. The GPCRs examined included chemokine receptors, serotonin receptors, adrenergic receptors, and histamine receptors. The docking results demonstrated high binding affinity between phytochemical drug structures and these receptors for most drug-receptor combinations.The study consists of three main parts. First, data on receptors and drugs were retrieved from the Protein Data Bank (PDB) and PubChem, respectively. The receptors selected are those predicted to be linked to Long COVID. The structure of the drugs was converted from SDF to PDB format using OpenBabel from ChemInfo. Docking studies were performed to assess the binding affinity between the drugs (ligands) and their respective receptors (binding sites) using iGemDock v2.1.In conclusion, the findings of this study could have significant implications for the development of alternative therapies for Long COVID.

One of the most crucial attributes of emulsions is their particle size, which has implication on the stability, rheology as well as sensory properties in different food applications. Homogenization speed is an important parameter that disrupts fat globules into desirable particle sizes. The effect of emulsification composition and homogenization parameters on particle size distribution of palm stearin oil-in-water emulsion was investigated. A palm stearin oil-in-water emulsion was homogenized at different speed (10k-25k rpm) and the particle size distribution was investigated. When the homogenization speed was increased above >10k rpm, the particle size distribution decreased. Increasing concentration of surfactant from 1 wt.% to 3wt.%, in the emulsion showed a decrease in particle size distribution for all homogenisation speed. Oil droplet size could also be minimized by increasing the concentration of sugar in continuous phase.

We investigated the cub’s survival in a wild Indian Leopard (Panthera pardus fusca) population in the Jhalana Forest Reserve, India. The research focuses on the survival analysis of leopard cubs during their first two years of life and determining the critical period of the reproductive cycle that influences survival. Survival functions were estimated using the Kaplan-Meier method using data collected with trail cameras over four years from 2018 to 2021. We found that the mean survival probability of cubs during the 2-year period they depend on their mother was 0.618, the first year – 0.739, and higher during the second year of life (0.831). The results also indicate that the first 12 months of a leopard cub's life are crucial for survival. These findings allow for understanding critical periods in the life history of individuals and are essential for effective conservation strategies in fragmented habitats.

Abscisic acid (ABA) and gibberellic acid (GA3) are regulators of fruit color and sugar levels, and the application of these hormones is a common practice in commercial vineyards dedicated to the production of table grapes. However, the effects of exogenous ABA and GA3 on wine cultivars remains unclear. We investigated the impact of ABA and GA3 applications on Malbec grapevine berries across three developmental stages. We found similar patterns of berry total anthocyanin accumulation, induced by both treatments, closely associated with berry H2O2 levels. Quantitative proteomics revealed that ABA and GA3 positively modulated antioxidant defense proteins, mitigating H2O2. Consequently, proteins involved in phenylpropanoid biosynthesis were downregulated, leading to decreased anthocyanin content at almost ripe stage, particularly petunidin-3-G and peonidin-3-G. Additionally, we noted increased levels of the non-anthocyanins E-viniferin and quercetin in the treated berries, which may enhance H2O2 scavenging at almost ripe stage. Using a linear mixed-effects model, we found statistical significance of the fixed effects berry H2O2 and sugar contents, demonstrating their roles in anthocyanin accumulation. In conclusion, our findings suggest a common molecular mechanism by which ABA and GA3 influence berry H2O2 content, ultimately impacting anthocyanin dynamics during ripening.

The ovary is the major determinant of female reproductive health. Ovarian functions are mainly related to the primordial follicle pool, which is gradually loss with aging. Ovarian aging and reproductive dysfunctions share oxidative stress as common underlying mechanism. ROS signaling is essential for normal ovarian processes yet can contribute to various ovarian disorders when disrupted. Therefore, balance in the redox system is crucial for proper ovarian functions. In the present study, by focusing on mRNAs and ncRNAs described in the ovary and taking into account only validated ncRNA interactions, we built up an ovarian antioxidant ceRNA network, namely OvAnOx ceRNA, composed of 5 mRNAs (SOD1, SOD2, CAT, PRDX3, GR), 10 miRNAs and 5 lncRNAs (XIST, FGD5-AS1, MALAT1, NEAT1, SNHG1). Our bioinformatic analysis indicated that the components of the OvAnOx ceRNA not only contribute to antioxidant defence, but are also involved in other ovarian functions. Indeed, antioxidant enzymes encoded by mRNA of OvAnOx ceRNA operate within a regulatory network that impacts ovarian reserve, follicle dynamics, and oocyte maturation in normal and pathological conditions. The OvAnOx ceRNA represents a promising tool to unravel the complex dialogue between redox potential and ovarian signalling pathways involved in reproductive health, aging and diseases.

Dogs and cats have become the most influential and successful pets through long-term domestication. People keep them for various reasons, such as their functional roles or for physical or psychological support. However, why humans are so attached to dogs and cats remains unclear. A comprehensive understanding of the current state of human preferences for dogs and cats and the potential influential factors behind it is required. Here, we investigate this question using two independent online datasets and anonymous questionnaires. We find that current human preferences for dogs and cats are relatively higher than for most other interests, such as games and music. Genetic variations, gender, age, and economic development levels notably influence these preferences for dogs and cats. Specifically, dog and cat ownership is significantly heritable (h2 = 0.43, 95%CI: 0.35 -0.51) and further analysis found that women, young people, and those with higher incomes are more likely to keep dogs and cats, and the primary reason is to gain emotional support. Our study provides insights into why humans get so attached to dogs and cats and establishes a foundation for developing co-evolutionary models.

Long COVID, characterized by persistent symptoms lasting from weeks to several months following initial infection with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), significantly impacts individuals' quality of life and poses a potential long-term burden on global healthcare systems. Clinical manifestations of Long COVID are diverse, affecting multiple systems including respiratory, cardiovascular, neurological, and gastrointestinal. While the precise mechanisms underlying Long COVID remain unclear, emerging evidence suggests involvement of various bodily pathways. This study employs bioinformatics tools to investigate the mechanisms of Long COVID and identify potential drug targets associated with key pathways, including inflammation, cytokines, immune response, coagulation, mitochondrial function, oxidative stress, and autonomic dysfunction. Tools such as the National Center for Biotechnology Information (NCBI) database, PubChem, UniProt, and iGEMDOCK were utilized for pathway analysis and molecular docking. The analysis aimed to identify drug molecules that may interact with target proteins to regulate or inhibit pathway activity, yielding several potential drug candidates for further investigation.

The Asiatic apple leafminer, Phyllonorycter ringoniella (Matsumura), is a significant secondary pest of apple trees in Northeast Asia. To better understand its population dynamics, a population model based on temperature-developmental relationships was constructed. This model includes three sub-models: spring emergence, immature stage transition, and adult oviposition. Field data were collected from sex-pheromone baited traps in apple orchards in Andong, Korea, dur-ing 2015 and 2016 to validate the model. Simulations under six pesticide-natural enemy scenari-os showed that the population size of each generation was best simulated when weighted mor-tality factors for pesticides and natural enemies were applied. Using daily temperature inputs, the model demonstrated that P. ringoniella typically undergoes five generations per year, with peak times predicted within a seven-day margin of field data. Sensitivity analyses revealed that population size was influenced by total fecundity and the larval stage model, but peak times remained consistent despite parameter changes. Higher temperatures led to earlier adult peak dates, especially in summer generations. This model serves as a fundamental tool for estimating population dynamics and abundance changes of P. ringoniella and can guide the timings of pesti-cide application. Further validation is necessary to test the model's efficacy in controlling pests in apple orchards.

During HIV-1 virus assembly the genomic RNA (vRNA) is selected for packaging by the viral protein Gag because it contains a specific packaging signal, Psi. While there have been numerous studies of Gag-Psi interactions, there is almost no information on kinetic aspects of this interaction. We have investigated the kinetics of Gag binding to different RNAs using switchSENSE DRX2 technology. We measured the association rate of Gag binding to monomeric Psi, to a “Multiple Binding Site Mutant” Psi that is inactive for genome packaging in vivo, and to a scrambled Psi. We discovered that Gag binds more rapidly to Psi RNA than to the mutant or scrambled RNAs. Furthermore, rapid Gag association kinetics are retained within sub-regions of Psi: Gag associates more rapidly with RNA containing only the 3’ two of the three Psi stem-loops than with monomeric RNA containing the 5’ two stem-loops or a scrambled RNA. No differences were detectable with individual Psi stem-loops. Interestingly, the rate of binding of Gag molecules to Psi increases with increasing Gag concentration, suggesting cooperativity in binding. The results are consistent with the hypothesis that selectivity in packaging derives from kinetic differences in initiation of particle assembly.

Enzyme catalysis has traditionally been used by various human cultures for creating value, long before its basic concepts have been uncovered, by preparing useful products through the trans-formation of raw materials available from natural resources. Tremendous scientific and techno-logical progress has been accumulated globally in understanding what constitutes an enzyme, what reactions enzymes can catalyze, and how to search, develop, apply and improve enzymes to make desired products. The exquisite properties of enzymes as nature's preferred catalysts, such as high selectivity, diversity and adaptability, enable their optimal work, whether in single reactions or in multiple reactions. Excellent opportunities for resource efficient manufacturing of compounds needed are provided by the actions of enzymes working in reaction cascades and pathways within the same reaction space, like molecular robots along a production line. Enzyme catalysis plays an increasing role for industrial innovation and responsible production in various areas, such as green and sustainable chemistry, industrial or white biotechnology. Sources of inspiration can be current manufacturing or supply chain challenges, the treasure of natural enzymes or the opportunities of engineering tailor-made enzymes. Making best use of the power of enzyme catalysis is essential for changing the way how current products are manufactured, how renewable biobased resources can replace fossil-based resources, and how the safety, health environment aspects of manufacturing processes can be improved towards cleaner and more sustainable production.

Measuring cell confluence in high-throughput screening of pharmaceutics, cell culture monitoring, cancer research, and industrial applications requires quantitative and standardizable measures to minimize subjective observations and increase throughput1-12 . Over the past half-century, non-invasive specialized programs have been created to quantitatively assess cell confluence, and some software tools exist to meet these needs1,4,5,6,7,8,9 . However, many of these tools are expensive, computationally-demanding, or inaccessible to non-experts1,6. With the assistance of generative artificial intelligence, we have developed the Confluence Analysis Program (CAP), a modern tool designed to be precise, interactive, and user-friendly. CAP is written entirely in Python and runs in any command-line interface (CLI), including Terminal (OS) and Command Prompt (Windows). In addition to fetching all necessary external libraries automatically and prompting drag-and-drop image retrieval, CAP features live visual comparison of cell mass identification to the original image assay. Our results demonstrate robust performance of CAP on fluorescent micrographs of varying confluences ranging from low to high. CAP is provided as a free resource, accessible in the Supplementary Materials.