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Β-Gals are a subgroup of the glycoside hydrolase (GH) family of enzymes, which possess the Glyco_hydro_35 (GH35) domain. Although studies have been conducted on the β-Gal gene family in numerous plant species, no such research has been conducted on beans. The purpose of this study was to determine the gene expression levels of β-Gal genes in the leaf tissue of P. vulgaris under salt and drought stress using quantitative real-time polymerase chain reaction (qRT-PCR) and to perform a comprehensive analysis of β-Gal gene family members using bioinformatics tools. In the bean genome, 25 Pvul-βGAL proteins with amino acid numbers ranging from 291 to 1119, molecular weights from 32.94 to 126.56 kDa, and isoelectric points from 5.46 to 9.08 were identified. Both segmental and tandem duplication have occurred in β-Gal genes in the bean genome, and Pvul-BGAL genes have been subject to negative selection in the evolutionary process. For a deeper comprehension of the evolutionary proximity of Pvul-BGAL genes, a phylogenetic tree and synteny map were drawn together with Arabidopsis thaliana and Glycine max β-Gal genes. The expression profiles of β-Gal genes in different tissues of the bean were determined in silico. In addition, the expression profiles of β-Gal genes in the leaves of bean plants subjected to drought and salt stress were analyzed, and the role of β-Gal genes in salt and drought stress was estimated. In this study, the role of β-Gal gene family in abiotic stress response and the characterization of β-Gal genes in beans were determined for the first time and will provide a basis for future functional genomics studies. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is a widespread chronic bone and joint disease for which there is currently no effective preventive or therapeutic treatment. Accumulating evidence indicates that circular RNAs (circRNAs), a class of noncoding RNAs, play critical roles in OA. Therefore, in this study, we aimed to reveal an unexplored circTBCK and elucidate its mechanism of action in the pathological process of OA. The different expression of circTBCK was obtained both in vitro and in vivo. In the in vivo model, mice were induced via destabilization of the medial meniscus (DMM) surgery, while in vitro model, mouse cells like primary chondrocytes of newborn mice and ATDC5 cell line were treated with IL-1β treatment (10 ng/mL for 24 h). The level of circTBCK was examined by quantitative real-time polymerase chain reaction (qRT-PCR). After circTBCK was overexpressed or knocked down, IL-1β treatment was performed, and then, chondrocyte viability was detected via a Cell Counting Kit-8 (CCK-8) assay at 0, 24, 48, or 72 h. To assess type II collagen (Collagen II) expression, immunofluorescence (IF) analysis was used. The levels of mRNAs and proteins related to proliferation, the extracellular matrix (ECM) and autophagy were determined by qRT-PCR and Western blotting. Compared with OA treatment, primary chondrocytes with treatment of both circTBCK overexpression and IL-1βincreased the expression of anabolic factors—Collagen II and SRY-box transcription factor 9 (SOX9), proliferation-related molecules—Ki-67 and proliferating cell nuclear antigen (PCNA), and autophagy-related molecules—Microtubule-associated protein 1 light chain 3 (LC3), B-cell lymphoma 1 (Bcl1), and autophagy-related 5 (Atg5) and decreased Sequestosome 1 (SQSTM1 or P62). In contrast, knockdown of circTBCK aggravated the chondrocyte degeneration induced by IL-1β. Overall, our findings suggest that circTBCK, an unexplored circRNA, could regulate autophagy, proliferation, and the extracellular matrix (ECM) to mitigate the development of OA, suggesting a possible target for OA prevention and therapy. [ABSTRACT FROM AUTHOR]

Potato (Solanum tuberosum L.), an important horticultural crop is a member of the family Solanaceae and is mainly grown for consumption at global level. Starch, the principal component of tubers, is one of the significant elements for food and non-food-based applications. The genes associated with biosynthesis of starch have been investigated extensively over the last few decades. However, a complete regulation pathway of constituent of amylose and amylopectin are still not deeply explored. The current in-silico study of genes related to amylose and amylopectin synthesis and their genomic organization in potato is still lacking. In the current study, the nucleotide and amino acid arrangement in genome and twenty-two genes linked to starch biosynthesis pathway in potato were analysed. The genomic structure analysis was also performed to find out the structural pattern and phylogenetic relationship of genes. The genome mining and structure analysis identified ten specific motifs and phylogenetic analysis of starch biosynthesis genes divided them into three different clades on the basis of their functioning and phylogeny. Quantitative real-time PCR (qRT-PCR) of amylose biosynthesis pathway genes in three contrast genotypes revealed the down-gene expression that leads to identify potential cultivar for functional genomic approaches. These potential lines may help to achieve higher content of resistant starch. [ABSTRACT FROM AUTHOR]

Papaya (Carica papaya L.) is a commercially important fruit crop cultivated worldwide due to its nutritional and medicinal values. Papaya ringspot disease (PRSD), caused by the papaya ringspot virus (PRSV), poses a significant threat to papaya cultivation, resulting in substantial yield losses. In this study, two independent field experiments were conducted at Bagalkote located in the Northern Dry Zone of Karnataka state of India. The first experiment aimed to identify the optimal planting month for papaya to effectively manage PRSV disease. The second experiment was conducted to determine the susceptible papaya growth stage for PRSV infection. The results revealed that early planting of papaya in June or late planting in March were identified as the most optimal planting times across the majority of growth stages, as they exhibited the lowest disease incidence along with superior growth and yield characteristics compared to other planting months. In contrast, planting during the winter season (September to January) resulted in high disease severity due to an increased aphid population. Conversely, planting during periods of low aphid activity (spring season) delayed disease onset until the monsoon. By the time the aphid population increased, the plants had already progressed beyond the flowering and fruit-bearing stages. In the second experiment, the severity and frequency of foliar symptoms on PRSV-inoculated papaya plants were significantly higher in those inoculated at the early growth stage compared to those inoculated at a later growth stage. This indicates that the early growth stage (up to 180 days after transplanting) is a critical period for PRSV infection, necessitating the implementation of effective disease management measures during this time to minimize disease spread and enhance growth and yield. Furthermore, plants inoculated at the early stage exhibited a higher viral titer, more severe symptoms, and a higher percent transmission rate compared to those inoculated at a later stage. These findings were supported by qRT-PCR analysis, which demonstrated a highly significant and positive correlation between early inoculation and disease severity. [ABSTRACT FROM AUTHOR]

Introduction: The basic helix-loop-helix (bHLH) proteins are a large family of transcription factors that are essential to physiology, metabolism, and development. However, the available information is limited about the bHLH gene family in Chimonobambusa utilis , which is widely cultivated in China because of its high-quality and economic value. C. utilis cultivars exhibit five natural color variations in their shoot sheaths, but the molecular mechanism behind this color diversity remains unclear. Methods: De novo assembly was employed to obtain gene sequences. To identify pathways related to color formation, GO enrichment analysis was performed on the 44,255 functionally annotated unigenes. Results: The transcriptomic analysis of C. utilis yielded a total of 195,977 transcripts and 75,137 unigenes after removing redundancy. The enrichment results revealed that four pathways were most strongly associated with color formation. Phylogenetic, conserved motif, and protein–protein interaction analyses, along with qRT–PCR validation, confirmed CubHLH17 's role in red sheath color. Discussion: This research not only deepens insights into the functional roles of CubHLH genes but also lays the foundation for genetic improvement of bamboo species. We suggest that these findings will contribute to both scientific research and commercial bamboo cultivation through gene editing technology in the future. [ABSTRACT FROM AUTHOR]

AbstractIntroductionMethodsResultsDiscussionThis study aimed to assess the expression changes of BTG1, PGI, and PGII in tissues and serum of patients with gastric cancer, atrophic gastritis, and healthy individuals.QRT-PCR was used to measure BTG1, PGI, and PGII expression in 30 cancers, 30 atrophic gastritis, and 30 healthy tissue samples. Serum levels of PGI and PGII were measured using ELISA. Statistical tests included the Mann–Whitney U and independent T-test. Covariates like tumour stage and H. pylori status were considered.BTG1 expression was significantly lower in cancer and gastritis tissues. Serum PGI and PGII levels were significantly reduced in cancer patients (P ≤ 0.001).The PGI/PGII ratio in serum emerged as a strong non-invasive biomarker for distinguishing cancer from healthy individuals. While BTG1 provides insights into gastric carcinogenesis, its clinical utility is limited due to the need for tissue samples. The serum-based PGI/PGII ratio shows greater promise as a non-invasive screening tool for GC. [ABSTRACT FROM AUTHOR]

Background: The transition from vegetative to floral development is a critical process regulated by a variety of complex internal and external factors. This study aims to characterize the Flowering Locus D (FLD) gene in relation to the floral induction pathway and examine its expression profiles under different photoperiodic conditions in Indian bean (Lablab purpureus (L.) Sweet). Methods: The LprFLD gene was PCR-amplified and sequenced using gene-specific primers for both photosensitive (GNIB-22, GP-189) and photoinsensitive (GNIB-21) varieties of Indian bean. Exon prediction, phylogenetic analysis, and in silico domain prediction were performed, followed by gene expression analysis using qRT-PCR for contrasting phenotypes. Results: The complete LprFLD gene sequence was characterized, containing five exons and showing significant similarity to GmFLD and VuFLD. Phylogenetic analysis of FLD homologs from 27 taxa revealed the closest evolutionary relationships among members of the Phaseoleae tribe, including Lablab purpureus, Vignaumbellata, Vigna angularis, Vignaradiata, Vigna unguiculata, Phaseolus vulgaris, Glycine max, Glycinesoja, and Cajanus cajan. In silico domain prediction identified two conserved domains, SWIRM and amino oxidase, in the FLD homologs of Indian bean, soybean, cowpea, Arabidopsis and rice. Gene expression analysis showed that the relative expression of LprFLD in trifoliate leaves corresponded with flowering in both photosensitive and photoinsensitive genotypes under long-day conditions. Overall, the expression level of LprFLD was lower in trifoliate leaves and shoot tips. Conclusion: This is the first report on the characterization of the LprFLD locus in Indian bean. Phylogenetic analysis confirmed its close evolutionary relationship with other Phaseoleae species. qRT-PCR results suggest that the elevated expression of LprFLD in trifoliate leaves coincides with flowering time in both photosensitive and photoinsensitive genotypes under long-day conditions, indicating that other loci may also regulate floral transition under varying photoperiods in Indian bean. [ABSTRACT FROM AUTHOR]

TCP transcription factors are a unique class of transcription factors that play important roles in alleviating abiotic stresses such as drought and salt. In this study, the whole-genome data of three cultivated varieties, namely, "SFS", "Sang" and "OT3098v2", were utilized to identify and analyze the members of the TCP gene family in oats, and their responses to two abiotic stresses, drought and salt, were also investigated. Results showed that there are 83, 65, and 30 non-redundant TCP genes in the three oats, with the highest number of TCP genes specific to the "SFS", reaching 22 genes. The oat TCP genes can be classified into three subfamilies: PCF, CIN, and CYC/TB1. Most AsTCP genes have important motifs, Motif 1 and Motif 2, which are part of the bHLH domain. Additionally, various cis-acting elements related to hormone response, abiotic stress, light response, and growth and development were found in the promoters of AsTCP genes. The main amplification mechanism of the oat TCP gene family is fragment duplication. Two tandem duplications, AsTCP058 / AsTCP059 and AsTCP023 / AsTCP025 , are stably present in the three oats. The highest number of AsTCP collinear relationships exist in the "SFS" with 89 pairs. After drought and salt stress treatments, significant differences in gene expression were observed among different oat cultivars and treatment periods. Genes that showed significant expression changes under both treatments (AsTCP021 , AsTCP033 , AsTCP044 , AsTCP053 , and AsTCP058) may play important roles in oat's response to abiotic stresses. Notably, AsTCP053 gene was significantly upregulated at 24 h of stress treatment and showed a more sensitive response to salt stress. This study provides insights into the functional characterization of the oat TCP gene family and its molecular mechanisms underlying stress tolerance. [ABSTRACT FROM AUTHOR]

The aim of this study was to deeply explore the pathogenesis of obesity type 2 diabetes mellitus (O-T2DM) and search for potential biomarkers through high-throughput RNA sequencing technology. The study included 15 patients with O-T2DM and 15 healthy controls, and peripheral blood samples were collected for transcriptome analysis. The results showed that compared with the control group, there were 442 circRNAs and 2756 mRNAs with significant differential expression in the O-T2DM group. Through weighted gene co-expression network analysis (WGCNA) and pathway enrichment analysis, it was found that the differentially expressed mRNAs were mainly enriched in signaling pathways such as T cell receptor, cell senescence, cytotoxicity mediated by NK cells, IL-17, lipids and atherosclerosis, and the oxidative phosphorylation pathway was activated, and apoptosis was inhibited. Based on the ceRNA theory, a regulatory network was constructed, and key circRNAs such as hsa_circ_0060614 were screened out, which may regulate the expression of the MT2A gene by adsorbing hsa-mir-4668-3p, and the expression levels of the three were significantly increased in O-T2DM patients. This study provides a new perspective for the research on the molecular mechanism of O-T2DM and an important theoretical basis for the development of personalized treatment and precision medicine for it. [ABSTRACT FROM AUTHOR]

Background: Monkeypox virus (MPXV) has emerged as a significant global health concern with outbreaks worldwide. While MPXV is primarily known for its dermatological and systemic manifestations, it can also cause central nervous system (CNS) complications. This systematic review describes the demographic, clinical, diagnostic, and therapeutic characteristics of MPXV-associated CNS neuroinflammatory disorders. Methods: We systematically reviewed the literature to identify cases of MPXV-associated CNS neuroinflammatory disorders. Data on demographics, systemic and neurological manifestations, diagnostic methods, treatment strategies, and outcomes were extracted and analyzed. Results: Eighteen cases of MPXV-associated neuroinflammatory disorders were identified. The mean age of patients was 27.8 years (range: 28 days to 43 years), with a male predominance (66.7%). Diagnosis included The most common diagnoses were acute disseminated encephalomyelitis in nine cases (50.0%), encephalitis/meningoencephalitis in seven cases (38.9%, isolated transverse myelitis in one case (5.6%), and transverse myelitis with encephalitis in one case (5.6%). The latency between the onset of systemic symptoms and neurological involvement averaged 6.2 days. MPXV detection was confirmed in 13 of 18 (72.2%) cases, primarily using quantitative real-time polymerase chain reaction from various biological specimens. Among the 12 cases with documented treatment, the most commonly administered therapies were tecovirimat (58.3%) and intravenous methyl-prednisolone (66.7%). Outcomes were reported in 17 cases, with complete recovery in 29.4%, partial recovery in 41.2%, and death in 29.4% of patients. Conclusions: MPXV-associated neuroinflammatory disorders of the CNS are rare but cause significant complications. The findings underscore the need for clinical vigilance, advanced diagnostic approaches, and targeted therapeutic strategies. Further research is essential to elucidate mechanisms underlying MPXV neurovirulence and develop effective treatments for these life-threatening conditions. [ABSTRACT FROM AUTHOR]

Background: Quantitative Real-time PCR (qRT-PCR) is a powerful technique to analyze gene expression patterns by measuring the relative abundance of mRNA transcription levels. The most crucial step in obtaining accurate results of qRT-PCR is to select suitable reference genes. Water temperature is an important factor that affects various physiological processes of fish. Presently, Japanese flounder is a commercially important marine culture species and the study of its gene expression is increasing rapidly. However, the reference genes used for Japanese flounder in previous studies, especially under temperature stress, only focused on those well-known genes widely reported in vertebrates, which might not be the proper reference genes. Results: In this study, we evaluated the suitability of eight genes including ribosomal protein L6 (rpl6), ribosomal protein L9 (rpl9), delta (4)-desaturase, sphingolipid 1 (degs1), cathepsin L (ctsl), eukaryotic translation elongation factor 1 gamma (eef1g), NSA2 ribosome biogenesis homolog (nsa2), eukaryotic translation initiation factor 3, subunit E, a (eif3ea), glutamine amidotransferase class 1 domain containing 1 (gatd1) analyzed from RNA sequencing (RNA-Seq) data and two genes including β-actin (actb) and 18S rRNA ribosomal RNA (18S RNA) selected from literature to obtain the best internal controls in qRT-PCR analysis of Japanese flounder under temperature stress. The statistical analysis methods (delta-Ct, BestKeeper, geNorm, and NormFinder) were further used to determine candidate reference gene stability. Initial results showed the suitability of eight genes from RNA-Seq data, which exhibited more stable expression levels than two commonly reported reference genes. Further analysis revealed that gatd1 and rpl6 were the best reference genes in Japanese flounder exposed to temperature stress. Conclusion: This study transcriptome-wide identified reference genes in different tissues of Japanese flounder exposed to temperature stress for the first time, providing a basis for gene expression research in flatfish. [ABSTRACT FROM AUTHOR]

Background: One of the primary challenges that the expanding population faces is water scarcity. Thus, a global imperative has been established to safeguard extant water resources and optimize their utility through sustainable practices and efficient management. In the present investigation, Azolla pinnata, a pteridophyte (fern), was employed to phytoremediate Cr (VI) from chromium-polluted water. The potential of this treated water for agricultural purposes was verified through the use of Vicia faba plants. Results: In vitro, A. pinnata effectively remediates Cr (VI) from an array of liquid concentrations (0.05 to 90 ppm) in a ratio of 25:1 {volume (mL): fresh weight of Azolla (g)} after 2 days incubation period at room temperature. At low concentrations (0.1 ppm), the phytoremediation capacity peaked at 70%, falling to 19.53% at a high concentration (90 ppm). Upon continuous irrigation with Cr-polluted water (0.05 to 50 ppm), the in vivo pot experiment on Vicia faba plants revealed high Cr accumulation in the roots reached 52.5 mg Kg-1 dry weight (Dwt) at the 50 ppm Cr treatment. Nevertheless, a reduced Cr content of 19.5 mg Kg-1 Dwt was observed when the plants were irrigated with 50 ppm Cr-polluted water that had been treated with Azolla. At 50 ppm of Cr, Azolla's treatment significantly increased shoot length, fresh weight, and Chl a content to 25.25 cm, 3.4 g, and 6.5 mg g-1 Dwt, respectively, up from 10.25, 1.8, and 4.7 in untreated plants. The chromosomal aberrations were significantly induced in the dividing cells of all Cr treatments, with the highest value of 4.8% at 50 ppm. This value was reduced to 2.88% at the same concentration when treated with Azolla. At a concentration of 10 ppm Cr, the mitotic index was significantly improved to 6.99% when combined with Azolla, as opposed to 3.63% when the same concentration was used without Azolla. The DNA degradation assay showed partial DNA degradation at 50 ppm Cr, which the Azolla treatment eliminated. Furthermore, the gene expression levels of both the PM H+-ATPase and the calcium-dependent protein kinase CDPK5 were upregulated in response to Cr, despite the fact that the expression level was altered in a dose- and concentration-dependent manner by Azolla treatment. Conclusion: Azolla exhibits substantial potential for reducing the detrimental effects of chromium stress including oxidative stress on plants. It modulates stress-related gene expression, protects DNA integrity, enhances cell mitosis, and reduces chromosomal damage. These results indicate that Azolla has the potential to be a valuable asset in phytoremediation strategies for chromium-contaminated environments, and that it may enhance plant survival and growth under Cr stress conditions. Key message: Azolla pinnata can be effectively utilized as an environmentally-friendly method to remediate chromium-contaminated water for agricultural usage. Highlights: • A. pinnata remediates Cr(VI) from liquid concentrations with 70% phytoremediation capacity at low concentrations. • A. pinnata treatment greatly decreased Cr buildup and improved Vicia faba growth under Cr stress. • In Vicia faba, A. pinnata increased mitotic index, reduced Cr-induced chromosomal aberrations, and modulated the expression of genes linked to stress. [ABSTRACT FROM AUTHOR]

Feline infectious peritonitis virus (FIPV) remains as one of the leading causes of morbidity and mortality in young cats from shelters and catteries worldwide. Since little is known about the molecular characteristics of currently circulating FIPV strains in Long Island, New York, samples from two shelter cats submitted to the Pathology Diagnostic Services of the Long Island University College of Veterinary Medicine, with gross and microscopic lesions consistent with those of FIP were processed for virus isolation, molecular characterization and full-length genome decoding. The younger shelter cat, a 1-year-old male (A15) was found dead without previous signs of illness. Postmortem examination revealed gross and microscopic lesions characterized by vasculitis, necrosis, hemorrhage, and pyogranulomatous inflammation confined to the colon and associated lymph nodes. The second cat, a 7-year-old spayed female (A37) had an identical clinical history and similar but widespread lesions, including fibrinous peritoneal effusion, cecal, colonic, renal, and hepatic involvement. The gross and microscopic diagnosis of FIP in these cats was confirmed by immunohistochemistry (IHC) demonstration of feline coronavirus antigen using mouse anti-FIPV3-70 monoclonal antibody. Virus isolation from saved frozen kidney and colon tissue was performed through several subsequent blind passages in MDCK and Vero cell lines. Confirmation of the FIPV isolation was done through qRT-PCR, IFA, western blot using N protein antibodies, and NGS of the full-length genome sequencing. The full-length genome sequences of the virus isolate from the two cats were decoded using next-generation sequencing (NGS) and deposited in the GenBank as accession numbers PQ192636 and PQ202302. The genome size of these isolates was (29355 and 29321) nucleotides (nt) in length, respectively. While their genome organization was consistent with other FIPV genomes as follows (5'UTR-ORF1ab-S-3abc-M-E-7b-3'UTR-3'), marked differential mutations were observed in the ORF1a/b, S, 3Abc, and 7b protein genes of the two FIPV isolates. One notable deletion of 34 nucleotides was observed in the 7b genes of one of these isolates but was absent in the other. We confirmed the potential recombination events during the evolution of those two FIPV field isolates with the potential parent virus as FECoV-US isolated in 1970 and the potential minor parent as the Canine coronavirus. Our results provide a comprehensive molecular analysis of two novel FIPV isolates causing fatal disease in shelter cats from Long Island. Diagnostic surveillance with molecular characterization and sequencing analysis of circulating FIPV strains within animal shelters may help early detect unique emerging clinical and pathological manifestations of the disease and develop more targeted prophylactic and therapeutic approaches to control it. [ABSTRACT FROM AUTHOR]

Key message: Melatonin increases Pb tolerance in P. ovata seedlings via the regulation of growth and stress-related phytohormones, ROS scavenging and genes responsible for melatonin synthesis, metal chelation, and stress defense. Lead (Pb) is a highly toxic heavy metal that accumulates in plants through soil and air contamination and impairs its plant growth and development. Because of its pharmaceutical importance, improvements in Plantago ovata yield against abiotic stresses are necessary. Melatonin (MEL) is a stress-alleviating biostimulator and our results showed a reduction in Pb induced phytotoxicity by enhancing plant growth attributes and balancing protective osmolytes. Pb-induced reactive oxygen species accumulation, including superoxide and peroxide free radicals and their mitigation through enzymatic antioxidants, was demonstrated in presence of MEL. Cell viability and Pb bioaccumulation were determined to understand the extent of cellular damage. Moreover, MEL increased secondary metabolite (flavonoids and anthocyanins) contents by 2–3-fold at the lowest Pb concentrations. Similar increases in the relative expression of genes (PoPAL and PoPPO), which are responsible for the production of non-enzymatic antioxidants, were observed. Notably, the upregulation of the PoCOMT gene up to 4-fold indicates increased melatonin production, as manifested in the phytomelatonin level. MEL supplementation also increased the auxin (IAA) level by 3-fold in the 100 µM Pb treatment group, while the abscisic acid (ABA) level decreased (1.4-fold) and the expression of PoMYB (a stress-related transcription factor) increased (up to 2.66-fold). Additionally, we found extreme downregulation (up to 18-fold) in the relative expression of PoMT 2 (a metal binding thiol compound) with melatonin treatment, which is otherwise upregulated (by 6-fold) during Pb stress. In the current study, these effects collectively revealed that MEL contribute to enhanced plant growth and Pb stress tolerance. [ABSTRACT FROM AUTHOR]

The transcription factor Tbx20 is integral to heart development and plays a significant role in various cardiac diseases. Despite its established importance, the regulatory mechanisms and functional significance of Tbx20 remain incompletely understood. To elucidate these mechanisms, we initially conducted eQTL mapping to identify genetic loci associated with Tbx20 expression in heart tissue from BXD mice. Co-expression and enrichment analyses revealed pathways linked to Tbx20, including dilated cardiomyopathy, hypertrophic cardiomyopathy, and FoxO signaling. Additionally, protein–protein interaction studies identified essential cardiac proteins, such as Myl2 and Myl7, along with upstream regulators like Mef2c. To validate our bioinformatic findings, we performed quantitative reverse transcription polymerase chain reaction (qRT-PCR) to assess the relative mRNA expression levels of TBX20 and Mef2c in the heart tissues of BXD mice compared to their parental strains (B6 and D2). Our results demonstrated significant up-regulation of both TBX20 and Mef2c in the BXD group relative to the parental strains. Conversely, both genes were down-regulated in B6, D2, Control, and Treatment groups when compared to BXD mice. These findings confirm the predicted regulatory roles of TBX20 and Mef2c in cardiac development as suggested by our initial analyses.This study not only reinforces the critical role of Tbx20 in cardiac gene regulation but also highlights its potential as a therapeutic target for cardiovascular disorders. Further investigations into Tbx20 and its interactions will enhance our understanding of heart biology and contribute to the development of targeted therapies for heart diseases. [ABSTRACT FROM AUTHOR]

Herbicide residues can negatively impact crop rotation and plant growth-promoting bacteria. This study examined the effect of atrazine on the expression of genes related to IAA production and phosphate solubilisation in Bacillus stercoris B.PNR1 and B.PNR2 under in vitro conditions. Results showed that B.PNR2 exhibited a more significant upregulation of the genes encoding aldehyde dehydrogenase (locus tags NZD81_05285 and NZD81_18505) and glucose 1-dehydrogenase (locus tags NZD81_05445 and NZD81_17730) compared to B.PNR1 when exposed to 200 µg/L of atrazine in the culture medium (P < 0.05). Specifically, the expression levels of NZD81_05445 and NZD81_17730 were over 30 times higher at 200 µg/L compared to the control (P < 0.05). The growth of mung bean seedlings in atrazine-contaminated soil varied by inoculation method. Seeds soaked in bacterial cell suspension for 24 h had a low survival percentage (41.25% to 51.25%), while directly pouring the suspension into the soil or soaking root-protruded seeds resulted in better survival (75.00% to 94.29%). The expression of both genes may be associated with enhanced growth of mung bean seedlings in atrazine-contaminated soil, as B.PNR2 has demonstrated a more effective trend than B.PNR1 in both soil conditions. For instance, shoot lengths of mung beans in non-contaminated and atrazine-contaminated soil were 16.35 ± 0.23 cm and 12.49 ± 0.21 cm with B.PNR2, compared to 14.94 ± 0.26 cm and 12.47 ± 0.21 cm with B.PNR1. Applying B.PNR2 directly to the soil resulted in specific root lengths of mung bean seedlings grown in atrazine-contaminated soil measuring 3.14 ± 0.06 m/g. In contrast, B.PNR1 produced a specific root length of 2.86 ± 0.06 m/g. Both bacterial suspensions demonstrated significantly higher specific root lengths than the control group, which measured 1.84 ± 0.05 m/g (P < 0.05). [ABSTRACT FROM AUTHOR]

Background: Oxidative damage has been implicated in multiple neurodegenerative diseases, including epilepsy. Selenium, in the form of selenoproteins is an integral part of the human antioxidant defense system. Though a relationship between the altered selenium levels and epilepsy has been reported, limited evidence is available about the expression pattern of selenoproteins in epileptic patients. Objective: This study aimed to determine the serum selenium levels in idiopathic epileptic and healthy individuals. Expression profiling of selenoproteins (GPx1, TRxR1 and SEPW1) both at mRNA and protein levels was also evaluated. Methods: Serum selenium levels of 30 patients with idiopathic generalized epilepsy and their age and gender matched 30 healthy controls were measured. Protein levels of Serum Glutathione Peroxidase 1 (GPx1), Thioredoxin Reductase 1 (TRxR1) and Selenoprotein W (SEPW1) were estimated using ELISA. mRNA expression of GPx1, TRxR1 and SEPW1 were determined using qRT-PCR. Results: The mean values for serum selenium levels in cases and controls were 37.6 ± 2.0 µmol/ml and 38.9 ± 2.7 µmol/ml, respectively. Selenium levels in cases were significantly lower as compared to controls (p = 0.031). No statistically significant differences were observed between the serum levels of selenoproteins GPx1, TRxR1 and SEPW1 in epileptic patients and the healthy group. GPx1 and TRxR1 expression was found to be down regulated (0.34 and 0.13 folds respectively) whereas SEPW 1 was found to be 0.04 folds up regulated in epileptic patients compared to the healthy subjects. Conclusion: Selenium deficiency observed in epileptic patients suggests the association between serum selenium levels and epilepsy. This study provides the information about the selenium status in Pakistani population and helps in understanding the role of selenium in the prevention of epilepsy. [ABSTRACT FROM AUTHOR]

Cortisol is the major glucocorticoid, which is considered to be a key factor linking the environmental stress and sex differentiation in teleosts. In the present study, we explored the possible role of cortisol in sex differentiation in a protogynous hermaphroditic swamp eel, Monopterus albus. We cloned and characterized the full-length cDNA sequences of glucocorticoid receptor genes, gr1 and gr2. A novel alternative splicing variant (gr1b) was also discovered, which lacked a 27-bp nucleotide insertion in gr1 , resulting in a nine-amino-acid deletion in GR1. Real-time PCR indicated that swamp eel gr1a mainly expressed in the brain; gr1b mainly in the spleen, head kidney, kidney and muscle; gr2 mainly in the muscle. During female-to-male sex reversal, the expression levels of gr1a and gr1b decreased significantly at the stage of ovotestis III, and gr2 showed a trend of increase, and increased significantly in the testis. In the brain, acute heat stress up-regulated significantly the expression of gr1a and gr1b , while had no significant effect on gr2. However, in the ovary, heat stress only up-regulated significantly the expression of gr1b at 12 h. Serum cortisol concentration increased significantly by heat stress at 1 h, and returned to normal levels at 6 and 12 h. Cortisol injection up-regulated significantly the expression levels of gr1b and gr2 , while had no significant effect on gr1a in the ovary. At the same time, cortisol injection down-regulated significantly the expression of cyp19a1a , and only decreased significantly the expression of foxl2 at 6 and 12 h. Cortisol injection up-regulated significantly the expression of dmrt1a at 6, 24 and 48 h, while only increased the expression of gsdf at 6 h. The results indicate that cortisol and GRs may play roles in sex differentiation by the regulation of gonadal differentiation-related gene expression in swamp eel. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant Klebsiella pneumoniae poses a severe risk to global public health, necessitating the immediate development of novel therapeutic strategies. The current study aimed to investigate the effectiveness of the green algae Arthrospira maxima (commercially known as Spirulina) both in vitro and in vivo against carbapenem-resistant K. pneumoniae. In this study, thirty carbapenem-resistant K. pneumoniae isolates were collected, identified, and then screened for their susceptibility to several antibiotics and carbapenemase production genes using PCR. Both blaKPC and blaOXA-48 genes were the most predominant detected carbapenemase genes in the tested isolates. The phytochemical profiling of A. maxima algal extract was conducted using LC–MS/MS in a positive mode technique. The minimum inhibitory concentrations (MIC) of the algal extract ranged from 500 to 1000 µg/mL. The algal extract also resulted in decreasing the membrane integrity and distortion in the bacterial cells as revealed by scanning electron microscope. The bioactive compounds that were responsible for the antibacterial action were fatty acids, including PUFAs, polysaccharides, glycosides, peptides, flavonoids, phycocyanin, minerals, essential amino acids, and vitamins. Moreover, A. maxima algal extract revealed an antibiofilm activity by crystal violet assay and qRT-PCR. A murine pneumonia model was employed for the in vivo assessment of the antibacterial action of the algal extract. A. maxima showed a promising antibacterial action which was comparable to the action of colistin (standard drug). This was manifested by improving the pulmonary architecture, decreasing the inflammatory cell infiltration, and fibrosis after staining with hematoxylin and eosin and Masson's trichrome stain. Using immunohistochemical investigations, the percentage of the immunoreactive cells significantly decreased after using monoclonal antibodies of the tumor necrosis factor-alpha and interleukin six. So, A. maxima may be considered a new candidate for the development of new antibacterial medications. [ABSTRACT FROM AUTHOR]

Background and aim: Acute myeloid leukemia (AML) is a remarkably complex malignancy; with considerable genetic, epigenetic, and phenotypic heterogenicity. Circ-RNAs are a novel class of non-coding RNA. They may influence leukemia development and offer exciting possibilities for targeted AML diagnosis and therapy. This study aimed to detect circ_0002232 and circ-VIM expression levels in AML patients and their relation to the clinicopathological characteristics and disease outcome to assess the prognostic potential of both circ-RNAs and achieve a new target therapy for the disease. Methods: Circ_0002232 and circ-VIM gene expressions were measured in 60 AML patients and 30 controls using qRT-PCR. Results: Circ_0002232 was significantly downregulated in our patients compared to controls (P value < 0.001). On the other hand, circ‐VIM was notably upregulated in our patients (P value = 0.005). Using ROC curve, circ_0002232 and circ-VIM biomarkers could distinguish AML patients from controls with AUC 0.847, 0.683 and P value < 0.0001, = 0.004 respectively. Patients with downregulated circ_0002232 were significantly younger than upregulated patients (p value = 0.003). In addition, downregulated circ_0002232 was significantly associated with decreased hemoglobin level and increased overall survival (OS). Regarding high circ-VIM expression in AML patients, it was significantly correlated with lacking complete remission and leukocytosis. Conclusion: Circ_0002232 and circ-VIM could be valuable diagnostic biomarkers to differentiate AML patients from healthy controls in clinical use. Circ-VIM expression may influence AML prognosis. Further research is needed to validate the clinical utility of circ_0002232 as a prognostic marker for OS in AML patients. [ABSTRACT FROM AUTHOR]

Developing melon genotypes with resistance to Fusarium oxysporum f. sp. Melonis‐(FOM) race1.2 is a major goal in any breeding program. In this study, we identified the role of 11 gene markers that contribute to polygenic resistance during the FOM1.2–melon interaction.qRT‐PCR analysis elucidated upregulation of candidate marker genes AMT, DXPR, Fom‐2, GLUC, GalS, GRF3, MLO, PRK, RuBlsCo, TLP and WRKY in resistant 'Shante‐F1' and 'Khatouni', and susceptible 'Shante‐T' and 'Shahabadi' at 7, 14 and 21 days post‐inoculation (dpi). We also studied changes in defence‐related enzyme activity: chitinase (CHI), β‐1,3‐glucanase (GLU) and peroxidase (POX) in melon roots.AMT, GLUC and DXPR transcripts were upregulatied in leaf and root tissues of the resistant 'Shante‐F1' and 'Shahabadi'. Transcript levels for GalS and GRF3 increased 6.77‐ and 6.83‐fold in roots of 'Shante‐F1' at 7 dpi, whereas in PRK, TLP and WRKY theye increased by 7.84‐, 5.15‐ and 12.26‐fold at 14 dpi, respectively. However, transcript levels increased by 5.18‐fold for Fom‐2 and 8.46‐fold for MLO at 21 dpi. Also, RBC transcript level peaked at 14 dpi with 4.9‐fold increase in leaves of resistant genotypes, whereas AMT increased 2.94‐fold at 21 dpi, and GLUC and DXPR increased 7.11‐ and 2.91‐fold at 14 dpi in 'Shante‐F', respectively. Defence‐related‐enzyme activity was also upregulated three‐fold in resistant varieties.The dynamic shifts in the melon transcriptome induced by FOM1.2 emphasize that resistance mechanisms are predominantly regulated through signalling pathways involving CHI, GLU, and POX defence response. Surprisingly, the AMT gene, basically resistant to downy mildew, Pseudoperonospora cubensis; GLUC, MLO and PRK resistant to powdery mildew (Sphaerotheca fusca); TLP and WRKY resistant to Phytophthora blight (Phytophthora capsici); and GRF3 and RBC resistant to root knot nematodes (Meloidogyne spp.) were upregulated in resistant genotypes, indicating a dual role of these genes in resistance to more than one disease at a time. [ABSTRACT FROM AUTHOR]

Key message: Fusarium oxysporum disrupts redox homeostasis in Vigna mungo, likely by interfering with salicylic acid signaling, which can be ameliorated by boosting PAL and its related pathways via salicylic acid pretreatment. Fusarium oxysporum, a widespread soil-borne fungus, significantly threatens global crops. This study centers on elucidating the infection strategies employed by F. oxysporum against a new and underexplored host Vigna mungo, a leguminous crop of high agronomic value, and the defense mechanisms that can be activated against the infection, aiming to uncover how these responses can be leveraged to develop potential countermeasures. Building on prior work demonstrating the in vitro antifungal efficacy of phytohormones, including salicylic acid (SA), this study further investigates SA pretreatment at 100 µM, which previously reduced reactive oxygen species (ROS) and improved germination under Fusarium stress. Through a comprehensive analysis of V. mungo plants pretreated with SA and subjected to F. oxysporum infection, we observed that fungal exposure reduced growth, chlorophyll content, and levels of proteins, phenolics and flavonoids, while increasing stress markers and antioxidant activity. SA pretreatment mitigated these effects by boosting antioxidant molecules and activating the phenylalanine ammonia-lyase (PAL) pathway, thereby enhancing endogenous SA and ROS scavenging. Furthermore, qRT-PCR analysis confirmed SA-mediated upregulation of antioxidant (catalase and peroxidase), fungal stress response genes ((pathogenesis-related gene 4 (PR4) and defensin (DEF)) and SA synthesis and regulator genes (PAL and WRKY70) involved in plant systemic resistance, while LC–MS data revealed an altered metabolic profile with increased phytoalexins and antioxidants synthesis. Overall, SA pretreatment confers resistance against F. oxysporum in V. mungo by modulating endogenous SA and metabolic profile to activate key defense pathways and redox homeostasis, highlighting its potential in plant defense strategies and reinforcing our proposed model of SA action. [ABSTRACT FROM AUTHOR]

Common beans hold significant importance in sustainable agriculture and their critical role in human nutrition cannot be overstated. Factors such as climate change underscore the necessity for expanding genetic diversity and delineating the attributes of local bean cultivars. Among the various abiotic stressors, drought emerges as a formidable constraint limiting bean cultivation. While diverse set of strategies are employed to mitigate the impacts of drought stress, the ultimate and enduring solution lies in the development of drought-resistant bean cultivars, and it can be stated that the same situation is valid for salinity stress. Flavonoid biosynthesis is very important for plant growth and development and flavonoids are known to be involved in many pathways including stress response. This study aimed to comprehensively identify and characterize the CHS gene family within different bean cultivars exposed to drought and salt stress, utilizing genome-wide analysis, and assessing gene expression levels. Employing a spectrum of in silico methodologies, 14 CHS genes were identified in the common bean genome. These genes exhibited molecular weights ranging from 37.38 to 43.34 kDa and consisted of 341–393 amino acid residues. Remarkably, all Pvul-CHS genes shared a common structure comprising two exons. Phylogenetic analyses conducted across Phaseolus vulgaris, Arabidopsis thaliana, and Glycine max revealed that the Pvul-CHS gene family could be classified into three primary clusters. The expression profiles of Pvul-CHS genes unveiled their varied tissue-specific expressions and their pivotal roles in responding to diverse abiotic stresses. Furthermore, we conducted in silico assessments of the chromosomal positions of CHS gene family members in beans, their orthologous associations with related genomes, and cis-acting element analyses. The outcomes of this study hold the potential to significantly contribute to the breeding of beans endowed with enhanced resilience against salt and drought conditions. [ABSTRACT FROM AUTHOR]

The N6-methyladenosine (m6A) methyltransferase METTL3 has been demonstrated to function in mediating m6A modification, but its role in ischemic stroke (IS) has not been fully elucidated. This study aimed to explore the downstream mechanism of METTL3-mediated m6A modification in IS. GSE16561 and GSE22255 were downloaded from the Gene Expression Omnibus database for analysis of differentially expressed genes (DEGs), and it was found that METTL3 mRNA was downregulated in IS. Then quantitative real-time polymerase chain reaction was used to verify the downregulation of METTL3 mRNA in the peripheral blood of IS patients and the cortexes of transient middle cerebral artery occlusion mice. By combining DEGs with the m6A-downregulated genes in GSE142386 which performed methylated RNA immunoprecipitation sequencing (MeRIP‐seq) on METTL3‐deficient and control endothelial cells, a total of 131 genes were identified as the METTL3-mediated m6A-modified genes in IS. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the genes were mainly involved in cytokine-cytokine receptor interaction, MAPK signaling pathway and NF-kappa B signaling pathway. CTSS and SBK1 were further screened as the key METTL3-mediated m6A-modified genes by random forest model and PCR validation. The ROC curve analysis showed that the combination with CTSS and SBK1 was of good diagnostic value for IS, with the AUC of 0.810, sensitivity of 0.780, and specificity of 0.773. Overall, we found that METTL3-mediated m6A modification may influence the occurrence and development of IS by participating in inflammation-related biological processes, and two key m6A-modified genes mediated by METTL3 (CTSS and SBK1) can be used as diagnostic biomarkers for IS. [ABSTRACT FROM AUTHOR]

Severe COVID-19 and MIS-C are rare but serious outcomes associated with SARS-CoV-2 infection. The onset of MIS-C often involves the gastrointestinal system, suggesting a potential connection with gut microbiota. This study aims to compare the gut microbiota of children with severe COVID-19 and those with MIS-C using various biomolecular approaches. Gut microbiota composition and specific microbial modulations were analyzed using fecal samples collected at hospital admission. The study included hospitalized patients (mean age 6 ± 5 years) diagnosed with severe COVID-19 (37 patients) or MIS-C (37 patients). Microbial differences were assessed using both NGS and qRT-PCR methodologies. In 75% of cases, pharmacological treatments included antibiotics and corticosteroids, which influenced the microbiota composition. Early age was found to have the most significant impact on microbiota diversity. Significant differences in alpha and beta diversity were observed between COVID-19 and MIS-C patients, particularly concerning low-abundance species. Levels of Bacteroides spp., Bifidobacterium spp., and Akkermansia muciniphila were comparable between groups, while an increased activity of Bifidobacterium spp. was noted in children with positive fecal samples (p = 0.019). An in-depth evaluation of lesser-known gut species may be key to reducing the risk of severe outcomes and developing microbiota-based biomarkers for the early diagnosis of MIS-C. [ABSTRACT FROM AUTHOR]

The use of antidepressant drugs during pregnancy is increasing globally. SNRIs and SSRIs are most widely used for treating panic disorders. Venlafaxine is an antidepressant that inhibits serotonin and norepinephrine reuptake. Given the increasing prevalence of antidepressant use during pregnancy, Venlafaxine exposure during the early stages of development could lead to changes in protein dynamics and disrupt essential gene pathways, raising concerns about the potential teratogenic effects of antidepressant exposure during pregnancy. This study explores the developmental impact of Venlafaxine, a commonly prescribed serotonin-norepinephrine reuptake inhibitor (SNRI), on protein biochemistry and gene expression in the developing Gallus gallus domesticus embryos. Significant changes in protein profiles were observed among control and Venlafaxine-treated groups. Sonic Hedgehog (SHH) gene, a key regulator of embryonic patterning and organogenesis, specifically neural tube formation, and limb development, is known for its crucial role in embryonic development. qRT-PCR analysis of Venlafaxine-treated embryos showed changes in SHH gene expression suggesting that Venlafaxine may target SHH gene expression, and potentially alter signalling pathways crucial for normal embryogenesis. However, detailed studies are needed to understand the long-term consequences of prenatal and foetal antidepressant exposure on embryonic growth and development. These changes may lead to abnormal growth patterns or congenital defects, emphasizing the need for caution when prescribing Venlafaxine during pregnancy. [ABSTRACT FROM AUTHOR]

Abstract The increasing temperature is a major threat to plant growth and development. It severely alters various biochemical and physiological processes and ultimately affects the overall crop yield. The membrane-based remorin protein-encoding genes (REM) were previously reported as significantly involved in the regulation of various biotic and abiotic stressors. However, these REM genes were not studied in Cajanus Cajan, the sixth most important legume crop due to its rich protein source and traditional medicinal plant. In this study, 17 REM gene orthologs were identified in C. cajan against A. thaliana REM genes and verified through the presence of conserved REM-related domains. Phylogenetic analysis revealed that REM genes were divided into six different groups. All the REM genes were unevenly distributed on 11 chromosomes of C. cajan. 3D protein structures and intron-exon organization indicate conserved evolutionary pattern within C. cajan. Various core, hormone-responsive, and stress-responsive cis-regulatory elements were found in promoter regions of REM genes, including TATA-box, CAAT-box, MYB, and G-box. The total estimation of antioxidant enzyme activity revealed the increase in POD and SOD activity, potentially due to a defense mechanism in response to high temperature. RNA sequencing data processing reveals higher expression of CcREM genes in leaf and flower, including CcREM1.1 and CcREM1.2. Furthermore, the differential change in expression was observed in response to high-temperature stress. Among these genes, one upregulated gene (CcREM1.3) and two downregulated genes (CcREM6.1 and CcREM6.5) are potential candidate targets for heat stress response, followed by qRT-PCR validation. Our findings suggest that CcREM1-like and CcREM6-like genes hold significant potential for future climate-smart heat-tolerant breeding of C. cajan.

This study was designed to identify the efficiency of interferon-stimulated gene 15 (ISG15) expression for early pregnancy detection in Awassi ewes. A total of 10 ewes were enrolled: 3 carrying twins, 4 carrying single pregnancies, and 3 non-pregnant. Blood samples were collected on 0, 6, 15, and 25 days of gestation to measure ISG15 expression levels using quantitative real-time PCR (qRT-PCR). The results of the relative expression values of ISG15 in pregnant and non-pregnant ewes on 0, 6, 15, and 25 days of pregnancy were 0.044±0.012 vs. 0.049±0.009, 0.203±0.083 vs. 0.201±0.093, 1.839±0.594 vs. 0.292±0.136, and 0.583±0.067 vs 0.200±0.051, respectively. In both twin and single pregnant ewes, the relative expression values of ISG15 were increased significantly (P