Protein Gene Research Articles

Genomic insights into drug resistance and virulence determinants in rare pyomelanin-producing clinical isolates of Acinetobacter baumannii.

Clinical isolates of multi-drug resistant Acinetobacter baumannii are a major cause of nosocomial infections, often attributed to the highly adaptable genome that helps it tothrive under environmental selection pressure. Here, we aim to provide genotypic-based surveys and comparative whole genome sequencing (WGS) analysis to explore the genomics of the rare pyomelanin-forming clinical isolates of A. baumannii from India. A total of 54 clinical isolates of A. baumannii obtained from two tertiary care hospitals were genotyped using repetitive sequence-based PCR(REP-PCR) for elucidating their molecular epidemiology, followed by their resistance profiling through the determination of minimum inhibitory concentration using the micro broth dilution method. The isolates' virulence and antibiotic-resistant determinants were detected by PCR screening, followed by biofilm quantification. Pyomelanin pigment produced by A. baumannii isolates was isolated and chemically characterized. Finally, WGS of three pigment-producing and one non-producing A. baumannii strains was performed to explore the factors contributing to their variability. REP-PCR genotyping identified around 8 clusters, with all isolates being multidrug-resistant (MDR). Pyomelanin-producing isolates were strong biofilm formers, characterized by the concurrent presence of 'pgaB, BfmR, BfmS, ompA, and cusE' biofilm-related genes. These pigmented strains belonged to ST2Pas and co-harbored blaOXA-23, blaADC-25, aph (3')-VIa, armA, aph (6)-Id, tet(B) and msr(E) genes. Thirteen common IS elements and biosynthetic gene clusters of arylpolyene, NI-siderophore, and NRP-metallophore were identified. Notably, genomic islands containing aminoglycoside 3'-phosphotransferase, oxidative stress, two-component response regulators, efflux pump-related, toxin-antitoxin protein, and virulence-related genes were also mapped by WGS. The pyomelanin-forming isolates were MDR and virulent. The elucidation of WGS analysis provided critical insights for understanding the epidemiology, virulome, and mobilome of rare pigment-producing A. baumannii strains.

CD2AP deficiency aggravates Alzheimer’s disease phenotypes and pathology through p38 MAPK activation

BackgroundAlzheimer’s disease (AD) is the most common form of neurodegenerative disorder, which is characterized by a decline in cognitive abilities. Genome-wide association and clinicopathological studies have demonstrated that the CD2-associated protein (CD2AP) gene is one of the most important genetic risk factors for AD. However, the precise mechanisms by which CD2AP is linked to AD pathogenesis remain unclear.MethodsThe spatiotemporal expression pattern of CD2AP was determined. Then, we generated and characterized an APP/PS1 mouse model with neuron-specific Cd2ap deletion, using immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay, electrophysiology and behavioral tests. Additionally, we established a stable CD2AP-knockdown SH-SY5Y cell line to further elucidate the specific molecular mechanisms by which CD2AP contributes to AD pathogenesis. Finally, the APP/PS1 mice with neuron-specific Cd2ap deletion were treated with an inhibitor targeting the pathway identified above to further validate our findings.ResultsCD2AP is widely expressed in various regions of the mouse brain, with predominant expression in neurons and vascular endothelial cells. In APP/PS1 mice, neuronal knockout of Cd2ap significantly aggravated tau pathology, synaptic impairments and cognitive deficits. Mechanistically, the knockout of Cd2ap activated p38 mitogen-activated protein kinase (MAPK) signaling, which contributed to increased tau phosphorylation, synaptic injury, neuronal apoptosis and cognitive impairment. Furthermore, the phenotypes of neuronal Cd2ap knockout were ameliorated by a p38 MAPK inhibitor.ConclusionOur study presents the first in vivo evidence that CD2AP deficiency exacerbates the phenotypes and pathology of AD through the p38 MAPK pathway, identifying CD2AP/p38 MAPK as promising therapeutic targets for AD.

Unveiling the shared genes between systemic sclerosis and lung cancer

The risk of lung cancer is significantly increased in patients with systemic sclerosis (SSc), yet the specific genes underlying this association remain unexplored. Our study aims to identify genes shared by SSc and lung cancer. We identified differentially expressed genes (DEGs) from SSc and lung adenocarcinoma (LUAD) datasets (SSc: GSE95065, LUAD: GSE136043) in the GEO database. We found shared genes by intersecting top genes in protein–protein interaction networks by the STRING database. The area under the ROC curve (AUC) was calculated for each shared gene in validation datasets (SSc: GSE231692; LUAD: GSE43458), identifying PRKG2 as the core shared gene. We used the UALCAN platform to assess PRKG2 expression in LUAD patients at various stages and lymph node metastasis states, and compared disease-free survival (DFS) between low and high PRKG2 expression LUAD groups. PRKG2 was overexpressed in A549 cells to study its impact on lung cancer cell proliferation and invasion in vitro. We identified seven shared genes (SCN7A, AGTR1, WIF1, PRKG2, LTF, AQP4, COL10A1), with the AUC for PRKG2 exceeding 0.93 in both diseases (SSc AUC = 0.973; LUAD AUC = 0.939). The PRKG2 expression levels of LUAD patients with different clinical stages and lymph node metastasis states were consistently lower than those observed in normal individuals. The DFS of LUAD patients in the high PRKG2 expression group was higher than that in the low expression group (p = 0.028). In vitro experiments confirmed elevated PRKG2 expression inhibits the proliferation and invasion of lung cancer cells. PRKG2 is one of the genes shared by SSc and lung cancer, affecting the proliferation and invasion of lung cancer cells.

Why the adventitious roots of poplar are so colorful: RNAseq and metabolomic analysis reveal anthocyanin accumulation in canker pathogens-induced adventitious roots in poplar.

This study revealed a substrate-level synthesis of pigment cyanidin-3-O-glucoside and the redirection of metabolomic flux in the flavonoid/anthocyanin biosynthesis pathway in poplar adventitious roots (ARs) induced by stem canker pathogens. Recently, we observed a novel allometry on poplar stems, with copious colorful adventitious roots (ARs) induced by fungal canker pathogens. Here, we reveal chemical, physiological, and molecular mechanisms of AR coloration in poplar-pathogens (Valsa sordida/Botrosphaeria dothidea) interaction system using our phloem girdling-inoculation system. Light-induced coloration in ARs: red/rosy under sunlight, and milky white under shading. Chemical and metabolomic analyses indicated that numerous (93 in all 110) and high relative intensities/contents of flavonoids metabolites (mainly including flavonols, flavones, and anthocyanins class) accumulate in red ARs, some flavones and anthocyanins metabolites all contribute to the color of poplar ARs, and cyanidin-3-O-glucoside is the most abundant colorant. Integrated analysis of metabolomic and transcriptomic analysis suggested that sunlight exposure redirected the metabolomic flux from the flavonoid biosynthesis pathway to the flavonols and flavones branch pathways, induced by the upregulation of FLS (flavonol synthase/flavanone 3-hydroxylase) and other structural genes. The anthocyanins metabolomic analysis and the downregulation of the ANS (anthocyanin synthase) gene illustrated a retard of metabolomic flux from leucoanthocyanidins to anthocyanidins. Metabolomic results and the upregulation of the gene BZ1 (Bronze 1, anthocyanin 3-O-glucosyltransferase) illustrated that sunlight triggered a rapid biosynthesis of anthocyanin metabolites in poplar ARs, which based on the substrate level of anthocyanidins. Transcriptomic and RT-qPCR analyses showed that transcriptional factor MYB113, HY5 (Elongated hypocotyl 5), and COP1 (Ring-finger protein CONSTITUTIVE PHOTOMORPHOGENIC1) genes positively regulated the expression of the flavonoid/anthocyanin biosynthesis structural genes (such as the BZ1, FLS and LAR gene) in both sunlight-exposed red ARs and white ARs after light-exposure, suggesting sunlight induces anthocyanins biosynthesis through the interaction between "MBW" complex and COP1-HY5 module. Moreover, one SPL gene (squamosa promoter-binding-like protein gene, target of miR156, and one component of miR156-SPL module) was down-regulated in sunlight-exposed poplar ARs, implying the biosynthesis flavonoid/anthocyanin be regulated at the posttranscriptional level. This study provides a potential AR experimental system for research on flavonoid/anthocyanin biosynthesis in tree species.

Knockdown of OsPHP1 Leads to Improved Yield Under Salinity and Drought in Rice via Regulating the Complex Set of TCS Members and Cytokinin Signalling.

Plant two-component system (TCS) is crucial for phytohormone signalling, stress response, and circadian rhythms, yet the precise role of most of the family members in rice remain poorly understood. In this study, we investigated the function of OsPHP1, a pseudo-histidine phosphotransfer protein in rice, using a functional genomics approach. OsPHP1 is localised in the nucleus and cytosol, and it exhibits strong interactions with all sensory histidine kinase proteins (OsHK1-6) and cytokinin catabolism genes. Our results demonstrate that OsPHP1 functions as a negative regulator of cytokinin signalling. Knockdown of OsPHP1 enhanced the expression of positive cytokinin signalling regulators, such as OsHKs and OsAHPs (authentic phosphotransfer proteins), while downregulating negative regulators, such as type-A response regulators (OsRRs) and cytokinin catabolism genes (CKXs). Furthermore, OsPHP1 negatively influences abiotic stress tolerance, as evidenced by the increased sensitivity of OsPHP1-OE (overexpression) lines to salinity and drought. In contrast, OsPHP1-KD (knockdown) lines showed enhanced stress resilience, with better photosynthesis, increased tiller and panicle production, higher spikelet fertility, and grain filling. The study demonstrates that OsPHP1 suppresses antioxidant and stress-responsive genes, exacerbating ion toxicity and reducing osmolyte accumulation, thereby impairing plant growth and yield under stress conditions. These findings highlight OsPHP1 as a critical modulator of plant responses to abiotic stress and suggest potential genetic targets for enhancing crop stress tolerance.

A novel approach for efficient co-expression of two foreign genes based on the reverse genetic system of Newcastle disease virus

Newcastle disease virus (NDV) is an ideal model for exploring the mechanisms of the virus; it is also an optimal vector for developing vector vaccines and for cancer therapy. A reverse genetic system of NDV Mukteswar strain controlled by eukaryotic cellular RNA polymerase II promoter was established by reverse genetics technology. Based on the reverse genetic system, an open reading frame of the enhanced green fluorescent protein (EGFP) gene be inserted between the P and M genes of the viral genome and flanked with the gene start (GS) sequence and gene end (GE) sequence to form an independent transcription unit. The rescued virus was amplified in specific pathogen-free (SPF) chicken embryos for 10 generations, and the results showed that the recombinant virus could stably express the exogenous gene for at least 10 generations. Efficient expression of two exogenous genes synchronously is essential for the development of NDV-based multivalent vaccine candidates. Explore the possibility of simultaneous and efficient expression of two exogenous genes based on NDV vector. In the present study, a recombinant virus with co-expression of EGFP and cherry fluorescent protein (CFP) inserted between the intergenic regions of the P/M gene as two independent transcription units was successfully rescued. The results showed that the two exogenous genes could be expressed synchronously and efficiently. The results of biological analysis of the expression efficiency of exogenous genes showed that the EGFP in recombinant viruses with two exogenous genes was slightly lower than that of recombinant viruses with one exogenous gene, but the expression efficiency of CFP in recombinant viruses with two exogenous genes was higher than EGFP in both viruses. These recombinant viruses have similar growth kinetics but with a little attenuation in virulence compared with parental viruses. In conclusion, these data indicated that this study successfully established the reverse genetic system of the NDV Mukteswar strain and achieved the purpose of efficient expression of two exogenous genes synchronously in a novel approach, laying the foundation for the development of multivalent vaccines or tumor therapeutics using NDV as a vector.

Trans-differentiation of Jdp2-depleted Gaba-receptor-positive cerebellar granule cells to Purkinje cells

The Jun dimerization protein (Jdp2) gene is active in mouse cerebellar granule cells and its protein product plays a crucial role in the formation of the cerebellum lobes through programmed cell death. However, the role of Jdp2 in cellular differentiation and pluripotency in the cerebellum, and the effect of the antioxidation reaction on cell plasticity, remain unknown. N-acetyl-l-cysteine (NAC) induced the early commitment of the differentiation of granule cell precursors (GCPs) to neurons, especially Purkinje cells, via the γ-aminobutyric acid type A receptor α6 subunit (Gabra6) axis; moreover, Jdp2 depletion enhanced this differentiation program of GCPs. The antioxidative effect of NAC was the main driving force of this decision toward the neural differentiation of the GCP population in the presence of Gabra6 in vitro. This implies that antioxidative drugs are effective agents for rescuing oxidative-stress-induced GCP damages in the cerebellum and commit this Gabra6-positive cell population toward differentiation into Purkinje cells.

Vitamin D binding protein and receptor prevalence in a large population with periodontitis: single nucleotide polymorphism and transcriptomic profiling

BackgroundThere is an ongoing controversy regarding the expression of vitamin D receptor (VDR) and binding protein (VDBP) genes, as well as their polymorphisms, in periodontitis. We examined eight single nucleotide polymorphisms (SNPs) and performed a transcriptome-level bioinformatics analysis to clarify their relationship with periodontitis.MethodsTo explore VDR and VDBP polymorphisms, 600 subjects were included, including 307 patients with chronic periodontitis (CP) and 293 healthy controls. Genomic DNA was extracted from peripheral venous blood collected from each subject. A MassARRAY system was used to detect SNPs, including rs1544410G/A (BsmI), rs2228570C/T (FokI), rs7975232G/T (ApaI), rs731236T/C (TaqI), rs739837G/T, rs9729G/T, and rs3847987C/A in the VDR gene, and rs7041A/C in the VDBP gene. Then, we analyzed transcriptome sequencing datas of gingival tissues from two single-cell transcriptome sequencing studies to identify differential expression profiles. The objective was to further explore the potential association between VDR gene and gingival tissues in individuals with CP.ResultsThe regression analysis model revealed a significant relationship between rs739837G/T (P = 0.04) and rs7041A/C (P = 0.03) polymorphisms and CP susceptibility. Subjects carrying the TT genotype of rs739837 showed a decreased risk of developing CP compared to those carrying the GG + GT genotype (OR = 0.53, 95% CI = 0.29–0.99). Individuals carrying the AC + CC genotype of rs7041 showed a reduced risk of developing CP compared to those with the AA genotype (OR = 0.70, 95% CI = 0.51–0.97). Furthermore, allele C of rs7041 was found to have a protective effect against periodontitis (P = 0.03, OR = 0.75, CI = 0.58–0.98). However, no association was found between CP susceptibility and six other 6 SNPs (rs1544410, rs2228570, rs7975232, rs9729, rs731236, and rs3847987). Differential levels of VDR transcription were observed in gingival tissues during CP.ConclusionsVDR genetic variability and transcriptional expression are significant factors affecting susceptibility to CP. These findings suggested that rs739837 TT in VDR and rs7041 A/C in VDBP may be protective against periodontitis.

Predicted breeding values for relative scrapie susceptibility for genotyped and ungenotyped sheep

BackgroundScrapie is an infectious prion disease in sheep. Selective breeding for resistant genotypes of the prion protein gene (PRNP) is an effective way to prevent scrapie outbreaks. Genotyping all selection candidates in a population is expensive but existing pedigree records can help infer the probabilities of genotypes in relatives of genotyped animals.ResultsWe used linear models to predict allele content for the various PRNP alleles found in Icelandic sheep and compiled the available estimates of relative scrapie susceptibility (RSS) associated with PRNP genotypes from the literature. Using the predicted allele content and the genotypic RSS we calculated estimated breeding values (EBV) for RSS. We tested the predictions on simulated data under different scenarios that varied in the proportion of genotyped sheep, genotyping strategy, pedigree recording accuracy, genotyping error rates and assumed heritability of allele content. Prediction of allele content for rare alleles was less successful than for alleles with moderate frequencies. The accuracy of allele content and RSS EBV predictions was not affected by the assumed heritability, but the dispersion of prediction was affected. In a scenario where 40% of rams were genotyped and no errors in genotyping or recorded pedigree, the accuracy of RSS EBV for ungenotyped selection candidates was 0.49. If only 20% of rams were genotyped, or rams and ewes were genotyped randomly, or there were 10% pedigree errors, or there were 2% genotyping errors, the accuracy decreased by 0.07, 0.08, 0.03 and 0.04, respectively. With empirical data, the accuracy of RSS EBV for ungenotyped sheep was 0.46–0.65.ConclusionsA linear model for predicting allele content for the PRNP gene, combined with estimates of relative susceptibility associated with PRNP genotypes, can provide RSS EBV for scrapie resistance for ungenotyped selection candidates with accuracy up to 0.65. These RSS EBV can complement selection strategies based on PRNP genotypes, especially in populations where resistant genotypes are rare.

Spike(s) protein gene microevolution of SARS CoV-2 virus in Bolivian Population

Objective: analyze the population gene structure and phylogeny of the S gene of the SARS CoV-2 virus of COVID-19 (+) patients from the Plurinational State of Bolivia and then correlate its phylogeny with the different waves of contagion. Methods: three SARS-CoV-2 samples obtained by nasopharyngeal swabs from positive COVID-19 patients were sequenced by Sanger sequencing. 488 sequences of Bolivian SARS-CoV-2 were downloaded from GISAID until September 25, 2023. The genetic structure and phylogeny were analyzed to correlate the presence of the different variants with the waves of contagion. Results: three (3) sequences of 623pb, 1557pb, and 3060pb of the S gene were obtained, and the last one was analyzed with the sequences downloaded from GISAID, obtaining a phylogenetic tree and a network of haplogroups that revealed the formation of clades and nodes that gave rise to variants of the six waves that Bolivian inhabitants faced. The Y508S mutation was identified as new since it was not identified in the CoVariants® and CoV-Glue® databases. Conclusions: the phylogeny, haplogroup network, and Person's correlation coefficient revealed the existence of a positive correlation between the microevolution of the analyzed fragment and the appearance of the different waves of contagion. Methods: Three SARS-CoV-2 samples obtained by nasopharyngeal swab from positive COVID-19 patients were sequenced by Sanger sequencing. 488 sequences of Bolivian SARS-CoV-2 were downloaded from GISAID until September 25, 2023. The genetic structure and phylogeny were analyzed to then correlate the presence of the different variants with the waves of contagion. Results: 3 sequences of 623pb, 1557pb and 3060pb of the S gene were obtained, the last one was analyzed with the sequences downloaded from GISAID, obtaining a phylogenetic tree and a network of haplogroups that revealed the formation of clades and nodes that gave rise to variants of the six waves that Bolivian inhabitants faced. The Y508S mutation was identified as new, since it was not identified in the CoVariants® and CoV-Glue® databases. Conclusions: The phylogeny, haplogroup network and Person's correlation coefficient revealed the existence of a positive correlation between the microevolution of the analyzed fragment and the appearance of the different waves of contagion.

Impact of prolactin treatment on enhancing the cellular responses of MCF7 breast cancer cells to tamoxifen treatment

Breast cancer remains one of the most challenging diseases to treat due to its heterogeneity, propensity to recur, capacity to spread to distant vital organs, and, ultimately, patient death. Estrogen receptor-positive illness comprises the most common breast cancer subtype. Preclinical progress is hampered by the scarcity of medication-naïve estrogen receptor-positive tumour models that recapitulate metastatic development and treatment resistance. It is becoming increasingly clear that loss of differentiation and increased cellular stemness and plasticity are important causes of cancer evolution, heterogeneity, recurrence, metastasis, and treatment failure. Therefore, it has been suggested that reprogramming cancer cell differentiation could offer an effective method of reversing cancer through terminal differentiation and maturation. In this context, the hormone prolactin is well recognized for its pivotal involvement in the development of the mammary glands lobuloalveolar tissue and the terminal differentiation that drives the production of the milk protein gene and lactation. Additionally, numerous studies have examined the engagement of prolactin in breast cancer as a differentiation player that resulted in the ablation of tumour growth and progression. Here, we showed that a pre-treatment of the estrogen-positive breast cancer cell line with prolactin led to a considerable improvement in the sensitivity of this cancer cell to Tamoxifen endocrine therapy. We also showed a favourable prognostic value of prolactin receptors/estrogen receptors 1 (or alpha) co-expression on breast cancer patients outcomes, and this co-expression is highly correlated with the well-differentiated breast tumour type. Our results revealed a fruitful aspect of the effects of prolactin in improving the responses of breast cancer cells to conventional endocrine therapy. Moreover, these findings further validated the ability of prolactin as a persuader of a more differentiated and less aggressive breast cancer phenotype. Hence, it suggested a potential implication of prolactin as a therapeutic candidate.

Coat-protein Gene Based Identification and Characterization of Yellow Mosaic Virus Infecting Blackgram in Northern Karnataka, India

Aims: Coat-protein (CP) based identification and characterization of yellow mosaic virus infecting blackgram. Place and Duration of Study: The study was carried out during Kharif 2023 at College of Agriculture, Vijayapur. Methodology: The whole genomic DNA was isolated from the leaf tissues of both healthy blackgram plants and plants infected with the yellow mosaic virus by using a modified CTAB method. Specific primers for YMV were tried to amplify coat protein region of blackgram YMV. Results: Sequence analysis revealed that the CP gene of the begomovirus under study shared 99.8 per cent similarity with MYMV Shivamogga isolate (OM106035.1) at the nucleotide level. When comparing the deduced amino acid sequences of individual proteins from YMV infecting blackgram at Vijayapur with those of other begomoviruses, the highest identity was found with the MYMV Shivamogga isolate (OM106035.1), showing 84.59 per cent similarity. A phylogenetic tree constructed based on the full-length coat protein gene sequence of MYMV, along with 37 other Geminivirus sequences, formed two major clusters in which MYMV-Vijayapur isolate appeared in Cluster I. Conclusion: The results of the current study, based on similarities in coat protein sequence at both the nucleotide and amino acid levels, as well as phylogenetic analysis, confirmed the prevalence of MYMV strain rather than MYMIV, HgYMV and DoYMV in blackgram in northern parts of Karnataka.

Single and dual RPA-CRISPR/Cas assays for point-of-need detection of Stewart's wilt pathogen (Pantoea stewartii subsp. stewartii) of corn and Maize dwarf mosaic virus.

Pantoea stewartii subsp. stewartii and Maize dwarf mosaic virus (MDMV) infections severely affect corn productivity worldwide. Rapid point-of-need diagnoses of quarantine pathogens P. stewartii subsp. stewartii and MDMV are required for early detection, timely disease management and ensuring phytosanitary regulations. Recombinase polymerase amplification (RPA) is an isothermal technique suitable for rapid diagnostics using minimally processed samples. Integrating CRISPR/Cas collateral activities with the RPA assays further enhances the specificity and sensitivity of molecular toolkits for diagnostic assays. RPA-CRISPR/Cas12a assay targeting the intergenic spacer region between capsular polysaccharide genes cpsA and cpsB of P. stewartii subsp. stewartii detected 1 × 10-6 ng DNA μL-1 using real-time fluorescence and blue light observation methods, and 1 × 10-4 ng DNA μL-1 using the lateral flow dipstick (LFD). Likewise, RPA-CRISPR/Cas13a assay detected MDMV coat protein (CP) gene with an ultrasensitive detection limit of 3.69 × 10-7 ng μL-1 using the real-time fluorescence and blue light observation methods, and 3.69 × 10-5 ng μL-1 using the LFD. The dual RPA-CRISPR/Cas assays detected both pathogens without compromising the speed and/or detection sensitivity of the single assays. The validated assays provide a useful and sensitive molecular tool for detecting two quarantine pathogens of maize within a minimal resource framework suitable for fast-tracking the containment strategies. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Heat alters diverse thermal tolerance mechanisms: An organismal framework for studying climate change effects in a wild bird

Abstract The ability to cope with heat is likely to influence species success amidst climate change. However, heat coping mechanisms are poorly understood in wild endotherms, which are increasingly pushed to their thermoregulatory limits. We take an organismal approach to this problem, unveiling how behavioural and physiological responses may allow success in the face of sublethal heat. We experimentally elevated nest temperatures for 4 h to mimic a future climate scenario (+4.5°C) during a critical period of post‐natal development in tree swallows (Tachycineta bicolor). Heat‐exposed nestlings exhibited marked changes in behaviour, including movement to cooler microclimates in the nest. They panted more and weighed less than controls at the end of the four‐hour heat challenge, suggesting panting‐induced water loss. Physiologically, heat induced high levels of heat shock protein (HSP) gene expression in the blood, alongside widespread transcriptional differences related to antioxidant defences, inflammation and apoptosis. Critically, all nestlings survived the heat challenge, and those exposed to milder heat were more likely to recruit into the breeding population. Early life but sub‐lethal heat may therefore act as a selective event, with the potential to shape population trajectories. Within the population, individuals varied in their physiological response to heat, namely in HSP gene expression, which exhibited higher mean and higher variance in heat‐exposed nestlings than in controls. Heat‐induced HSP levels were unrelated to individual body mass, or among‐nest differences in brood size, temperature, and behavioural thermoregulation. Nest identity explained a significant amount of HSP variation, yet siblings in the same nest differed by an average of ~4‐fold and individuals in the population differed by as much as ~100‐fold in their HSP response. This massive variation extends previous laboratory work in model organisms showing that heat shock proteins may harbour cryptic phenotypic variation. These results shed light on oft‐ignored elements of thermotolerance in wild birds at a critical stage of post‐natal development. By highlighting the scope of heat‐induced HSP gene expression and coupling it with a suite of organismal traits, we provide a framework for future testing of the mechanisms that shape species success in the face of change. Read the free Plain Language Summary for this article on the Journal blog.

ADAM10-mediated β-klotho degradation: a key to FGF21 resistance in senescent vascular endothelial cells.

Fibroblast growth factor 21 (FGF21) is a key regulator of metabolism and cardiovascular health. However, its upregulation in aging and age-related disorders suggests the presence of FGF21 resistance. This study aimed to elucidate the mechanisms underlying senescence-associated FGF21 resistance in human umbilical vein endothelial cells (HUVECs) and to explore potential therapeutic interventions. Transcriptomic analysis revealed a significant reduction in the number of FGF21-regulated genes in senescent HUVECs compared to young cells, indicating the onset of FGF21 resistance. In young HUVECs, FGF21 inhibited inflammatory cytokines and upregulated mitochondrial ribosomal protein genes. Conversely, senescent HUVECs showed a downregulation of β-klotho, an essential co-receptor for FGF21, at the cell membrane, concomitant with an upregulation of ADAM10, a protease involved in β-klotho degradation. These changes were also observed in aged mouse aortas. ADAM10 was shown to bind directly to β-klotho and promote its degradation, contributing to FGF21 resistance. Inhibition of ADAM10 or overexpression of β-klotho restored FGF21 responsiveness in senescent HUVECs. Moreover, pharmacologically high concentrations of FGF21 were effective in overcoming resistance and restoring its regulatory effects on senescent cells. These findings suggest that ADAM10-mediated degradation of β-klotho is a central mechanism in the development of FGF21 resistance, and that targeting the ADAM10/β-klotho axis could represent a novel therapeutic approach to restore FGF21 sensitivity. Pharmacological administration of FGF21 may hold promise in treating vascular aging and its associated cardiovascular pathologies.

Determination of Zucchini Yellow Mosaic Virus and Watermelon Mosaic Virus Infections in Cucurbit Production Areas of Çanakkale Province from Türkiye

Viral diseases are among the most significant challenges in protecting plants of the Cucurbitaceae family, with viruses from the Potyvirus genus, such as zucchini yellow mosaic virus (ZYMV) and watermelon mosaic virus (WMV), causing up to 100% yield losses under favorable conditions. Despite the importance of these viruses, there have been no previous studies investigating potyvirus diseases in Cucurbitaceae production areas in Çanakkale province. Consequently, the status of these diseases in the region remains unknown. This study aims to address this gap by analyzing the presence of potyviruses in Cucurbitaceae production areas in Çanakkale. In the 2021 production year, a total of 137 samples exhibiting virus and virus-like symptoms were collected from various Cucurbitaceae production sites in Çanakkale province and its districts. The samples were tested using RT-PCR with primer pairs specific to WMV and ZYMV. From the infected samples, seven isolates were selected for further analysis, and the coat protein (CP) genes were amplified and sequenced. The results revealed that WMV was detected as a single infection in 78 samples, ZYMV in one sample, and mixed infections of ZYMV+WMV were found in 39 samples, indicating that WMV is notably prevalent in Çanakkale. Bioinformatics analyses demonstrated that the Turkish WMV and ZYMV isolates share more than 90% similarity with other isolates in both the local samples and the GenBank database. Phylogenetic analysis further revealed that Turkish WMV and ZYMV isolates are closely related to each other. This is the first study to reveal the presence and phylogenetic relationships of ZYMV and WMV in cucurbitaceous plants in Çanakkale province of Türkiye.

Analysis of GFAP variants in UK Biobank suggests underdiagnosis or incomplete penetrance of adult-onset Alexander disease

BackgroundAlexander disease is an autosomal dominant leukodystrophy caused by heterozygous pathogenic variants in the glial fibrillar acidic protein (GFAP) gene. Although increasingly recognised, there is evidence that Alexander disease, particularly...

Identification of Viral Diseases and Influences on Yield and Quality of Angelica sinensis

Angelica sinensis is a perennial herbaceous species mainly cultivated in the Gansu, Yunnan, and Qinghai provinces of China, and its dried roots have been widely used for nourishing blood and harmonizing vital energy, largely relying on its bioactive compounds (e.g., alkylphthalides, polysaccharides, and flavonoids). In recent years, viral diseases have been suspected to be present in A. sinensis in field cultivation. In order to reveal the infection status and causes, a survey and the identification of viral diseases and their influence on the yield and quality of A. sinensis were conducted in four different counties of Gansu province. The results showed viral disease rates of ca. 21% to 37% for potato virus Y (PVY) and tomato mosaic virus (ToMV), as well as ca. 2.8- to 8.9-fold decreases in root yield on a unit-area basis; meanwhile, the contents of the main bioactive compounds (i.e., ferulic acid, ligustilide, and polysaccharides) were significantly lower in the virus-infected plants (VIPs) compared with the virus-free plants (VFPs); there were significant positive relationships of the viral disease rate with planting density and expression levels of the PVY-coat protein (CP) and ToMV-CP genes (p < 0.01). The above-mentioned observations indicate that it is necessary and urgent to take measures (e.g., controlling plant density, rational rotation, and using virus-free seedlings) to prevent the spread of plant viruses.

Zinc finger protein rotund is essential for wings and ovarian development by regulating lipid homeostasis in Locusta migratoria.

Cys2-His2-type zinc finger (C2H2-ZF) proteins are involved in diverse biological processes. In insects, the wing and ovarian development is crucial for reproduction and evolution, yet the physiological roles of C2H2-ZF proteins in these processes remain underexplored. Here, RNA-seq analyses identified C2H2-ZF protein genes that were differentially expressed during wing formation in Locusta migratoria. Among these, the gene encoding a C2H2-ZF protein Rotund (Rn) was highly expressed in the wing pads of fourth- and fifth-instar nymphs. RNA interference mediated knockdown of LmRn in nymph stages resulted in pronounced abnormalities with curled wings and reduced wing area. LmRn knockdown led to reduced expression of lipid transport-related genes during wing morphogenesis, significantly decreased triglyceride (TG) level. In addition, we also find that LmRn knockdown impaired ovarian development and oocyte maturation in female adults, with decreased expression levels of lipid synthesis-related genes, and vitellogenin genes (VgA, and VgB) in the fat body. Meanwhile, the number of lipid droplets, TG content, and protein levels in the ovaries were significantly decreased after LmRn was silenced. Together, our findings reveal that LmRn is essential for wing and ovarian development by regulating lipid homeostasis in locusts, offering a potential target for insect pest management.

Genome Sequencing and Comparative Genomic Analysis of Attenuated Strain Gibellulopsis nigrescens GnVn.1 Causing Mild Wilt in Sunflower

Gibellulopsis nigrescens, previously classified in the Verticillium genus until 2007, is an attenuated pathogen known to provide cross-protection against Verticillium wilt in various crops. To investigate the potential mechanisms underlying its reduced virulence, we conducted genome sequencing, annotation, and a comparative genome analysis of G. nigrescens GnVn.1 (GnVn.1), an attenuated strain isolated from sunflower. The genome sequencing and annotation results revealed that the GnVn.1 genome consists of 22 contigs, with a total size of 31.79 Mb. We predicted 10,876 genes, resulting in a gene density of 342 genes per Mb. The pathogenicity gene prediction results indicated 1733 high-confidence pathogenicity factors (HCPFs), 895 carbohydrate-active enzymes (CAZys), and 359 effectors. Moreover, we predicted 40 secondary metabolite clusters (SMCs). The comparative genome analysis indicated that GnVn.1 contains more CAZys, SMCs, predicted effectors, and HCPF genes than Verticillium dahliae (VdLs.17) and Verticillium alfalfae (VaMas.102). The core–pan analysis results showed that GnVn.1 had more specific HCPFs, effectors, CAZys, and secreted protein (SP) genes, and lost many critical pathogenic genes compared to VdLs.17 and VaMs.102. Our results indicate that the GnVn.1 genome harbors more pathogenicity-related genes than the VdLs.17 and VaMs.102 genomes. These abundant genes may play critical roles in regulating virulence. The loss of critical pathogenic genes causes weak virulence and confers biocontrol strategies to GnVn.1.