Transcriptional Analysis Research Articles

Identification of the BBX gene family in blueberry at different chromosome ploidy levels and fruit development and response under stress

BackgroundBlueberry (Vaccinium spp.) fruits are rich in flavonoids such as anthocyanins and have a high nutritional value. The zinc finger protein transcription factor B-box (BBX) plays important roles in plant growth and development, hormone response, abiotic stress, and anthocyanin accumulation. However, studies on the BBX family in blueberry are lacking.ResultsIn total, 83 VcBBX and 24 VdBBX genes were identified in tetraploid and diploid blueberry, respectively. A correlation was observed between the number of BBX genes in blueberry and chromosome ploidy. Gene loss and specific replication during blueberry evolution may lead to an imbalance of quantitative relationship between VcBBX and VdBBX genes. The analysis of transcriptome and quantitative reverse transcription–polymerase chain reaction data revealed that the expression pattern of BBX genes depended on the developmental stage of blueberry fruit. Gibberellin inhibited the expression of most VcBBX genes. Abscisic acid promoted the expression of some members of the BBX family. The expression levels of VcBBX15b4, VcBBX21a1, and VcBBX30a in blueberry leaves were significantly downregulated under blue light treatment, whereas that of VcBBX15c3 was significantly upregulated under red light treatment.ConclusionIn total, 83 VcBBX and 24 VdBBX genes were identified in 2 types of blueberries. Fruit development and transcription profiles under different stresses were analyzed. These findings will support further investigation of how BBX genes are involved in regulating hormone treatment and light stress during the growth and development of blueberry.

Alternating inverse modulation of xylem K+/NO3 - loading by HY5 and PIF facilitates diurnal regulation of root-to-shoot water and nutrient transport.

Diurnal light-dark cycles regulate nutrient uptake and transport; however, the underlying molecular mechanisms remain largely unknown. Transcription factor MYB59 and ion transporter NPF7.3 participate in root-to-shoot K+/NO3 - translocation in Arabidopsis. In this study, transcriptional analyses and western blotting experiments revealed the diurnal expression of the MYB59-NPF7.3 module. ChIP-qPCR and EMSA showed that transcription factors HY5 and PIF directly bind to the MYB59 promoter. Phenotype analyses and ion content measurement indicated that HY5 and PIF antagonistically control root-to-shoot K+/NO3 - translocation through the MYB59-NPF7.3 module. We found HY5 proteins accumulate in roots and repress MYB59 transcription during daytime, while PIF proteins promote MYB59 transcription in the dark. The expression levels of the NPF7.3 transcript and protein are gradually decreased during daytime, but increased at night. The enhancement of K+/NO3 - loading into the xylem mediated by NPF7.3 could increase root pressure at night, which maintained the root-to-shoot water/nutrient translocation. This study reveals a synergistic mechanism between light signaling and nutrient transport in plants, and defines a diurnal molecular switch of driving forces for root-to-shoot water/nutrient translocation.

The essential role of the hickory StMADS11 subfamily in flower organogenesis and flowering time in Arabidopsis.

The StMADS11 subfamily genes play a crucial role in regulating flowering time, flower development, and bud dormancy in plants. These genes exhibit functional differences between annual and perennial woody plants. In hickory (Carya cathayensis Sarg.), the specific roles of these genes in flowering regulation have not been elucidated. In this study, we identified five StMADS11 subfamily genes in the hickory genome, designated as CcSVP-like, CcAGL24-like1, CcAGL24-like2, CcJOINTLESS-like1, and CcJOINTLESS-like2, based on their clustering characteristics. Sequence analyses revealed distinct structural features in this subfamily, including differences in intron length, C domain, and conserved motifs. Transcript analysis indicated high expression levels of these genes in female flower buds, along with a notable seasonal expression pattern. Overexpression studies on Arabidopsis have demonstrated that the StMADS11 subfamily genes lead to various floral organ and pod anomalies. Specifically, overexpression of CcSVP-like resulted in delayed flowering, while overexpression of CcAGL24-like1, CcAGL24-like2, CcJOINTLESS-like1, and CcJOINTLESS-like2 promoted flowering. Protein interaction studies have shown that the StMADS11 subfamily proteins bind to the CcFUL-like protein. Notably, CcFUL-like, CcSVP-like, CcJOINTLESS-like1, and CcJOINTLESS-like2 proteins were able to bind to the CcSOC1-like promoter and suppress its expression. Our findings elucidate the distinct roles of the StMADS11 subfamily genes in flower development and timing, contribute to developing the current understanding of flowering regulation in hickory, and offer a foundation for further studies in perennial woody plants.

Protective effects and bioinformatic analysis of narciclasine on vascular aging via cross-talk between inflammation and metabolism through inhibiting skeletal muscle-specific ceramide synthase 1.

The senescence of smooth muscle is one of the independent risk factors in atherosclerosis progression in which the vascular inflammation plays an important role on vascular dysfunction. This study is designed to explore the novel vascular aging biomarkers and screen the potential molecular interventional targets through bioinformatic analysis. Transcriptional analysis was conducted based on the GSE16487 open access database, which included 15 human vascular tissue samples from two groups: young group (≤ 60 years old, n = 8) and aged group (≥ 75 years old, n = 7). There were 275 differential expression genes (119 upregulated and 156 downregulated genes) with minimum 1.5-fold change between two groups. 9 genes were mainly participated in inflammation-related signaling pathways, in which narciclasine was validated as the most effective candidate for modulation the ceramide synthesis. In vitro and animal study demonstrated that narciclasine reversed vascular aging by inhibiting skeletal muscle-specific ceramide synthase 1 (CerS1), reducing the ceramide level derived from CerS1, and improving fat deposition and circulating glycolipid metabolism. Narciclasine attenuates vascular aging and modulates the cross-talk between inflammation and metabolism via inhibiting skeletal muscle-specific ceramide synthase 1.

Improving consumer trust in digital health: A mixed methods study involving people living with chronic kidney disease.

To explore preferences, experience and trust in digital health in people living with chronic kidney disease (CKD), and tailor these findings towards solutions that may enhance uptake of digital health services. Mixed methods study, with cross-sectional survey and individual interviews with adults living with CKD attending specialist appointments at an Australian metropolitan hospital. Descriptive statistics and Wilcoxon matched-pairs test were used for survey responses and thematic analysis of interview transcripts, both reported on a theme-by-theme basis provided an overall understanding of trust in digital healthcare. Digital health is changing the way health services are provided, and our results demonstrate that despite limited familiarity, participants are open to learn and adapt to existing digital models of care. Limited exposure to technology may undermine trust in digital health, and telehealth can promote improvements in health literacy. Having the choice in healthcare modalities can promote trust, which can arise from trustful relationships with clinicians who demonstrate genuine interest in patient care. Participants expressed more concerns about sharing identity data than health data online and worry about fragmented information among providers. They preferred public health services due to distrust generated by the perceived risk of private sector data commercialisation. Building trust requires increasing awareness of digital health benefits, promoting positive experiences, improving digital literacy and ensuring interoperability and transparency in digital healthcare systems. People with CKD want to learn and benefit from digital health. Choice and open disclosure on data management and purpose are paramount to building trust.

Harnessing the spatial and transcriptional regulation of monoterpenoid indole alkaloid metabolism in Alstonia scholaris leads to the identification of broad geissoschizine cyclase activities

Monoterpene indole alkaloids (MIAs) are valuable metabolites produced in numerous medicinal plants from the Apocynaceae family such as Alstonia scholaris, which synthesizes strictamine, a MIA displaying neuropharmacological properties of a potential importance. To get insights into the MIA metabolism in A. scholaris, we studied here both the spatial and transcriptional regulations of MIA genes by performing a robust transcriptomics analysis of the main plant organs, leaf epidermis but also by sequencing RNA from leaves transiently overexpressing the master transcriptional regulator MYC2. These transcriptomic studies notably demonstrated that the first steps of the MIA pathway are successively distributed in the internal phloem associated parenchyma and epidermis, and that MYC2 exert a remarkable transcriptional effect by modulating the expression of around 1000 genes. By combining these distinct datasets, we initiated the search for MIA-related genes encoding CYP71, based on the similarity of expression compared to already known MIA genes. Transient expression of these candidates in Nicotiana benthamiana leaves and yeast notably led to the identification of a related isoform of rhazimal synthase (RHS) capable of converting the MIA precursor geissoschizine into akuammicine, strictamine and 16-epi-pleiocarpamine. Investigating its catalytic mechanism revealed that strictamine results from rhazimal deformylation and that a similar mechanism may also explain 16-epi-pleiocarpamine synthesis. This prompted us to rename these enzymes geissoschizine cyclase due to their capacity of cyclizing geissoschizine into three different MIA scaffolds and to form both C-C and C-N bonds. This identification thus illustrates the potential of integrating spatial and transcriptional regulation analysis for MIA gene identification.

Interferon regulatory factor 2 of red-spotted grouper (Epinephelus akaara): Insights into its transcriptional profiling, antiviral potential, and function in macrophage polarization.

Interferon regulatory factor 2 (IRF2) is a member of the IRF family that is specifically involved in diverse immune responses via interferon (IFN)/IRF-dependent signaling pathways. In this study, IRF2 of Epinephelus akaara (EAIRF2) was identified and characterized by evaluating its structural and functional properties. EAIRF2 showed the highest homology with IRF2 of Epinephelus coioides and clustered with teleosts in the phylogenetic tree. The highest level of EAIRF2 mRNA was found in the blood under normal physiological conditions. In the immune challenge experiment, significant transcriptional modulation of EAIRF2 upon lipopolysaccharide (LPS), polyinosinic: polycytidylic acid (poly I:C), and nervous necrosis virus (NNV) challenge were observed. The subcellular localization assay confirmed the role of EAIRF2 as a transcription factor by revealing its specific nuclear localization. To elucidate its functional implications in antiviral defense, EAIRF2 was overexpressed in fathead minnow cells, which were subsequently infected with viral hemorrhagic septicemia virus (VHSV). Notably, cells overexpressing EAIRF2 exhibited a significant reduction in the transcription of VHSV genes. Concurrently, the genes associated with the IFN/IRF signaling pathway were upregulated. Furthermore, the Hoechst and propidium iodide dual staining assay, water-soluble tetrazolium-1 (WST-1) assay, and transcriptional analysis of B-cell lymphoma 2-associated X protein (Bax)/B-cell lymphoma 2 (Bcl-2) indicated that EAIRF2 possesses anti-apoptotic properties during viral infection and poly I:C treatment. Additionally, EAIRF2 overexpression in murine macrophages induced M1 polarization and augmented relative marker gene expression. Collectively, these findings suggest that EAIRF2 is a pivotal immune-related gene, specifically implicated in the IFN/IRF-mediated antiviral defense mechanism, apoptotic signaling pathway, and activation of macrophage-mediated immune responses in Epinephelus akaara. The finding of this study enhances our understanding of IRF2's function in teleost immunity and presents potential avenues for developing therapeutic strategies against viral infections and other immune-related conditions in aquaculture species.

Characterization of tumor necrosis factor receptor-associated factor 2 (TRAF2) in red-spotted grouper (Epinephelus akaara): In vivo and in vitro investigation of its role in the regulation of antiviral immunity and cell death.

Tumor necrosis factor receptor-associated factor 2 (TRAF2) is a key adaptor molecule in tumor necrosis factor receptor signaling complexes, facilitating downstream immune-related signaling cascades. This study aimed to elucidate its function in teleost fish by characterizing the TRAF2 homolog of the red-spotted grouper (Epinephelus akaara, EaTraf2). The open reading frame of EaTraf2 encodes a putative protein of 520 amino acids, containing several characteristic domains of TRAF2. These structural features of EaTraf2 are conserved across diverse organisms, with a relatively higher sequence identity to TRAF2 orthologs from other bony fish. Transcriptional analysis demonstrated that EaTraf2 was ubiquitously expressed in all examined tissues, with the highest level observed in blood. Upon immune challenge, EaTraf2 expression significantly increased in the early stages of stimulation in both blood and spleen. Subcellular localization analysis revealed that EaTraf2 is predominantly localized in the cytoplasm. Overexpression of EaTraf2 in fathead minnow (FHM) cells resulted in elevated levels of interferon and inflammation-associated genes following viral hemorrhagic septicemia virus (VHSV) infection, along with reduced viral gene expression. This provided compelling evidence that EaTraf2 possesses antiviral properties. Furthermore, EaTraf2 demonstrated the ability to promote cell death induced by oxidative stress. Additionally, luciferase reporter assays revealed that EaTraf2 activates the NF-κB signaling pathway upon poly(I:C) stimulation and the Jun N-terminal kinase (JNK) signaling pathway in response to H2O2 treatment. Overall, our study elucidated the role of EaTraf2 in regulating innate immune responses and mediating stress-induced cell death. These findings enhance our understanding of TRAF2 in fish and may contribute to improved health management strategies in finfish aquaculture.

Pathogenic Deep Intronic PCSK1 Variant Causes Proprotein Convertase 1/3 Deficiency in a Family.

Proprotein convertase 1/3 (PC1/3), encoded by PCSK1, is expressed in neuronal and endocrine cell types, where it activates a number of protein precursors that play roles in energy homeostasis. Biallelic PCSK1 loss-of-function mutations cause a polyendocrinopathy; a total of 34 patients were reported. An infant with congenital malabsorptive diarrhea of all carbohydrates underwent exome sequencing (ES), with particular consideration of PC1/3 deficiency, but no mutations were found. The onset of obesity in the second year of life increased suspicion of PC1/3 deficiency in the proband, as well as in his equally affected cousin. Transcript analysis revealed minor amounts of an aberrant PCSK1 transcript containing intron 9 sequence and encoding a premature stop codon (p.Pro400Valfs*35). A deep intronic PCSK1 variant, NG_021161.1(NM_000439.5):c.1196+2681T>A, was found to segregate in the proband's family with the disease. A minigene approach demonstrated that the identified deep-intronic variant underlies pseudo-exon inclusion of the intron 9 sequence in the transcript. The characteristic phenotype of PC1/3 deficiency might require extended genetic testing to make a timely diagnosis.

Assessment of the effects of cadmium, samarium and gadolinium on the blue mussel (Mytilus edulis): A biochemical, transcriptomic and metabolomic approach.

Improving the understanding of how chemicals affect on organisms and assessing the associated environmental risks is of major interest in environmental studies. This can be achieved by using complementary approaches based on the study of the molecular responses of organisms. Because of the known chemical pressures on the environment, regulations on the content of some chemicals, such as cadmium, have been mostly completed. In contrast, the environmental toxicity of rare earth elements (REEs), which are widely used in industry, has only recently begun to receive attention. Here, we investigated the effects of cadmium, and two REEs, samarium and gadolinium, on marine mussels under laboratory exposures. We found that after an 8-day exposure at 500 µg/L, the metals were bioaccumulated by the mussels. Furthermore, samarium and gadolinium affected two oxidative stress biomarkers, GST and SOD. Lipidomic analysis showed that lipid content was modulated by the REEs, but not by cadmium. Interestingly, several compounds belonging to the phosphoinositide metabolism were more abundant, suggesting a pro-mitotic or cell survival response, while a higher abundance of cardiolipins after samarium exposure suggested an alteration of mitochondrial activity. Moreover, depending on the tissue and the metal considered, transcriptional analyses revealed an effect on metallothionein, hsp70/90, energy metabolism enzymes, as well as pro-mitotic transcript accumulation. Thus, this study sheds a new light on metal toxicity and in particularl REEs by highlighting the accumulation and toxicity of cadmium, samarium and gadolinium at 500 µg/L at different molecular levels, from gene expression to the lipidome of blue mussels.

Informing the development of transmission modelling guidance for global immunization decision-making: A qualitative needs assessment.

In recent years, mathematical transmission models have been increasingly used to support immunization program decisions and to measure the impact and cost-effectiveness of interventions. However, countries face expertise-and resource-related barriers that limit the use and application of modelled evidence to inform decisions. The World Health Organization (WHO) established an Immunization and Vaccines Implementation Research advisory committee subgroup in 2023 to support immunization decision-makers to effectively generate, translate and use such evidence for strategies, policies, and programs. This study supports this effort, detailing the needs of end-users to inform content and format of the guidance. Fifteen in-depth interviews were conducted with vaccination decision-makers and modelers from all six WHO regions and across low-, middle- and high-income countries. Interviews explored: (i) how modelling is understood and used; (ii) the challenges faced when using modelled evidence; (iii) the types of guidance that would be most useful to enhance the use of modelled evidence. Analysis of transcripts was guided by the framework method, which structures the analysis of qualitative data. Participants with modelling expertise used it firsthand, systematically, and often in an advisory capacity. Less experienced users, often in policy advisory roles, were less confident in their understanding of modelling and some did not use it at all. Decision-makers with little or no modelling experience cited a need for more information to help them understand the value of modelling in their context and many supported its potential. All participants saw a need for capacity strengthening and localised application to instil confidence in using modelled evidence. Those with less experience expressed a need for ongoing interactive engagement with knowledge brokers and training. Insights from this study are being integrated into the development of guidance by WHO. By considering the diverse challenges and needs of both experienced and inexperienced users of modelling, the guidance will support immunization strategy and policy by responding specifically to immunization decision-makers information needs.

Systemic TRPV4 inhibition worsens retinal response to acute intraocular pressure elevation in older but not younger mice.

Previous evidence showed that transient receptor potential vanilloid 4 (TRPV4) inhibition was protective of retinal ganglion cell (RGC) loss after chronic intraocular pressure (IOP) elevation in young animals. However, the role of TRPV4 in mechanosensing IOP changes in the aging eye is not well understood. This study compared the recovery of retinal function and structure after acute IOP elevation in 3- and 12-month-old mouse eyes with and without TRPV4 inhibition. We examined retinal TRPV4 expression in 2-month-old rodent eyes using immunohistochemistry and transcript analysis of isolated macroglia and RGCs. To modulate TRPV4, mice were treated daily with either vehicle or a TRPV4 antagonist (HC-067047 10 mg/kg) delivered intraperitoneally for 7 days before and 7 days after IOP elevation (50 mmHg for 30 minutes). Retinal function and structure were assessed using dark-adapted full-field electroretinography and optical coherence tomography, respectively. We showed that Müller cells strongly expressed TRPV4. Seven days after IOP elevation, RGC functional recovery was significantly poorer in older mice treated with TRPV4 antagonist compared with age-matched vehicle controls (-54 ± 7% vs. -24 ± 10%, p=0.046) and their younger TRPV4 antagonist-treated counterparts (-5 ± 5%, p<0.001). This study showed that there was an age-related deficit in RGC functional recovery from IOP elevation with TRPV4 inhibition.

Analysis of TNS3-203 and LRRFIP1-211 Transcripts as Oral Cancer Biomarkers.

A recent pan-cancer transcriptome analysis indicated differential activity of alternative promoters of genes TNS3 and LRRFIP1 in head and neck squamous cell carcinoma compared to non-cancerous tissue. The promoters upregulated in head and neck squamous cell carcinoma regulate expression of transcripts TNS3-203 and LRRFIP1-211. Our aim was to investigate the biomarker potential of TNS3-203 and LRRFIP1-211 transcripts in oral cancer, the most common type of head and neck cancer. An in silico approach was used to characterize the promoters and transcripts of interest. Relative expression of TNS3-203 and LRRFIP1-211 transcripts was evaluated by qRT-PCR in a group of 46 oral cancer patients in samples of cancer and adjacent non-cancerous tissue. TNS3-203 was significantly overexpressed in oral cancer compared with matched non-cancerous tissue, so this transcript can potentially be used as a diagnostic biomarker. There were no differences in LRRFIP1-211 level between analyzed tissues. None of the investigated transcripts has prognostic potential in oral cancer. The results obtained indicate the diagnostic potential of TNS3-203, but not LRRFIP1-211 transcript and its role in oral carcinogenesis.

518. Newborn immune transcriptome changes rapidly and is shaped by maternal SARS-CoV-2 infection and/or vaccination

Abstract Background SARS-CoV-2 infection during pregnancy is associated with adverse maternal effects, but its impact on infant’s immune development is not well defined. Using transcriptional profiles we analyzed the impact of SARS-CoV2 infection and/or vaccination in pregnant people and their infants longitudinally Transcriptome changes in pregnant people with SARS-CoV-2 infection and/or vaccination Modular analysis of pregnant people with SARS-CoV-2 infection and/or vaccination compared with healthy uninfected controls. Red dot: overexpression, blue dot: underexpression, white/empty space no difference vs controls Methods Multicenter observational study of SARS-CoV-2-infected and/or vaccinated pregnant people and their infants. Pregnant people (infected [n=91], vaccinated [n=42], infected-and-vaccinated [n=14], controls [n=22]) and their infants (n=81, 38,14, 11, respectively) were included. Maternal blood samples were collected during different trimesters and at delivery. Infant blood samples were collected longitudinally from birth (&amp;lt; 72 hours), at 1 week and 1, 3 and 6 months of age. Whole blood RNA was extracted for RNA-sequencing and transcriptomic data analyzed with R Transcriptional analysis of immune system development in early life Modular gene expression scores were analyzed longitudinally of individual infant samples. Red: higher expression, white: lower expression. D1-3: days 1 to 3, W1: week 1, M1: month 1, M3: month 3, M6: month 6 Results Compared with uninfected controls, interferon and plasma cells genes were overexpressed in pregnant people with acute infection (&amp;lt; 14 days), but underexpressed with earlier infection or vaccination during pregnancy (Figure 1). Newborns &amp;lt; 72 h of age (from all groups) showed significantly increased expression of erythrocytes, neutrophils, and inflammation genes. Expression of T/cytotoxic cell genes started to be observed at 1 week, while expression of B cells, plasma cells and interferon genes was observed at 3 months of age (Figure 2). Quantitative gene set enrichment analysis showed that compared to controls, infants from infected and/or vaccinated mothers had increased expression of interferon, while neutrophil, and adaptive immunity genes were decreased (Figure 3) SARS-CoV-2 infection and/or vaccination of pregnant people shaped immune profiles of their infants Fold changes of gene sets in infants compared to controls are shown (adjusted p-value &amp;lt;0.01) from quantitative gene set enrichment (QuSAGE) analysis. Red: overexpression, blue: underexpression. Infant groups are defined by maternal infection and/or vaccination status at delivery. Conclusion Interferon and plasma cells related pathways were overexpressed in pregnant people with acute COVID-19, while vaccination and SARS-CoV-2 infection earlier in pregnancy were associated with under-expression of both innate and adaptive immunity genes. The infant immune system changed rapidly in the first few days of life, as different immune programs followed separate time trajectories. Maternal SARS-CoV-2 infection and/or vaccination shaped the immune profiles of their newborns, suggesting a potential impact on their immune development Disclosures Rodrigo DeAntonio, MD, MSc, DrPH, GSK: Grant/Research Support|Moderna: Grant/Research Support|Sanofi: Grant/Research Support Asuncion Mejias, MD, PhD, MsCS, Astra-Zeneca: Advisor/Consultant|Astra-Zeneca: Honoraria|Enanta: Advisor/Consultant|Janssen: Advisor/Consultant|Janssen: Grant/Research Support|Merck: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Pfizer: Advisor/Consultant|Pfizer: Honoraria|Sanofi-Pasteur: Advisor/Consultant|Sanofi-Pasteur: Honoraria Octavio Ramilo, MD, Pfizer, Sanofi, Gates Foundation, NIH, and Merck Sharp &amp; Dohme LLC, a subsidiary of Merck &amp; Co., Inc., Rahway, NJ, USA (MSD): Advisor/Consultant|Pfizer, Sanofi, Gates Foundation, NIH, and Merck Sharp &amp; Dohme LLC, a subsidiary of Merck &amp; Co., Inc., Rahway, NJ, USA (MSD): Grant/Research Support|Pfizer, Sanofi, Gates Foundation, NIH, and Merck Sharp &amp; Dohme LLC, a subsidiary of Merck &amp; Co., Inc., Rahway, NJ, USA (MSD): Honoraria|Pfizer, Sanofi, Gates Foundation, NIH, and Merck Sharp &amp; Dohme LLC, a subsidiary of Merck &amp; Co., Inc., Rahway, NJ, USA (MSD): SAC member for MSD

Overexpression of OsDUF868.12 enhances salt tolerance in rice

Excessive salt accumuln in soil is one of the most important abiotic stresses in agricultural environments. The Domain of Unknown Function 868 (DUF868) family, comprising 15 members in rice, has been identified in the protein family database. In this study, we cloned and functionally characterized OsDUF868.12, a member of the OsDUF868 family, to elucidate its role in rice response to salt stress. A series of experiments, including RT-qPCR, Agrobacterium-mediated transient transformation in tobacco for localization analysis, phenotypic characterization, physiological and biochemical index measurement, and leaf staining, were conducted to investigate the function of OsDUF868.12 under salt stress. Transcriptional analysis revealed that OsDUF868.12 exhibited the most significant response to low temperature and salt stress. Preliminary subcellular localization studies indicated that OsDUF868.12 is localized in the cell membrane. Phenotypic Identification Experiments showed Overexpression lines of OsDUF868.12 enhanced resistance to salt stress and increased survival rates, while knockout lines of OsDUF868.12 were opposite. Physiological and biochemical assessments, along with leaf staining, demonstrated that overexpression of OsDUF868.12 improved the activity against oxidative stress.under salt stress. Furthermore, overexpression of OsDUF868.12 elevated the transcription levels of positively regulated salt stress-related genes. These findings suggest that overexpression of OsDUF868.12 enhances rice tolerance to salt stress at the molecular level through a series of regulatory mechanisms. This study provides valuable insights into the functional roles of the DUF868 family in plant responses to abiotic stress.

Exploring the clinical, psychological, and social relevance of menopause for trans and gender diverse people: a qualitative study.

The aim of this study was to explore the meanings and experiences of menopause for trans and gender diverse (TGD) people and how menopause affects clinical practice. For this qualitative study in 2021-2022, a scoping literature review informed interview schedule development. Following email invitations, online semi-structured interviews were conducted in March-April 2021 in Australia with three prominent TGD community leaders (trans male, trans female, nonbinary person) and three experienced medical practitioners (general practitioner, endocrinologist, psychiatrist), which were audio-recorded and transcribed. Inductive, iterative thematic analysis of transcripts was conducted. A further literature search was completed in 2024. Themes identified included the following: (1) no consensus on the definition of menopause for TGD individuals; (2) diverse menopause experiences existing in the TGD community; (3) large variations present regarding perimenopausal hormone therapy for older TGD patients: maintain dose, taper to low dose, or cease hormones; and (4) minimal support for TGD people to discuss menopause and concerns around aging. Saturation was not reached with the sample size. There is a paucity of relevant research and no consensus on definitions, management, or impacts of menopause for TGD people. Therefore, current management approaches should be individually tailored and guided by clinical expertise. To formulate practical and realistic recommendations for clinical practice, future research could include the following: (1) a broader literature review into varied gender-affirming hormone treatment (GAHT) regimens among aging TGD people; (2) utilizing the Delphi technique to achieve expert consensus regarding GAHT in older TGD individuals; and (3) a prospective study design of TGD participants on different GAHT regimens over time.

Conceptualizing the corporate-social accountability and sustainability assessment of weather-risk management policy towards stakeholders using qualitative insights: a case of weather index insurance

Purpose Sustainability assessment of risk-management policies has been needed to analyze their corporate social impact on stakeholders in terms of environmental, social and governance (ESG) and corporate social responsibility (CSR) accountability aspects. Thus, weather index insurance (WII) is a significant risk-management policy; there is a need to explore the corporate socio-environmental impact of WII on stakeholders. Thus, the purpose of this study is to develop a theoretical framework for theorizing the sustainability reporting aspect of WII. Design/methodology/approach This study performs qualitative exploratory research using in-depth interviews with ten experts. It has been carried out through semi-structured interviews with academic and professional experts and analyzed the interview transcripts using qualitative analysis. Findings This study’s outcomes contribute a new paradigm in the ESG and CSR accountability domain from the perspectives of stakeholder theory. The findings of this study reported a novel conceptual framework for integrated sustainability reporting of WII based on ESG and CSR constructs. The thematic content analysis of interview transcripts and WII literature exposed the significant accountability of WII towards human, ecological and financial capital. Practical implications This study provides valuable insights into the theoretical development of WII sustainability domain by developing a theoretical framework that provides a further pathway for future researchers. Social implications This study attempts to analyze the WII exposure toward ESG and CSR dimensions that are helpful for insurers, farmers, managers and governments. Originality/value This study firstly bridges the gap between WII theoretical knowledge and professional practice in terms of sustainability assessment of WII.

Genetic Services in Appalachia Conference Series.

This study examined the data generated as part of a seven-session webinar series that focused on genetics care provision in the medically underserved, rural Appalachian region and examined how these services have adapted to challenging practice environments. Barriers and facilitators to care in our region were considered. Data included a baseline survey of registrants, transcripts of sessions, and feedback about sessions. We analyzed data with a sequential and concurrent mixed methods approach. Registrants (n = 137) were disproportionately healthcare providers with genetic services expertise (37.2%). Approximately half (43.8%) of registrants were from KY, WV, and TN in the central Appalachian region. Our baseline survey found that the most noted barriers were the cost of services, lack of providers, and access to care. The most common facilitator was telehealth. Analysis of transcripts identified barriers that were consistent with those noted in the baseline survey, but additional support and network opportunities were discussed to allow for learning across services. Numerous barriers to service delivery were noted; however, despite challenges, participants identified opportunities and resources in the community. These insights will inform a research agenda aimed at advancing genetics services in rural Appalachia, addressing challenges, and leveraging assets for improved healthcare access and outcomes.

Establishing Reference Genes for Accurate Gene Expression Profiling in Toxigenic Bacillus cereus

Bacillus cereus is a Gram-positive pathogen associated with foodborne illnesses and severe non-gastrointestinal infections. Robust tools for accurate gene transcription analysis are essential for studying toxin gene expression dynamics and deciphering the complex regulatory networks orchestrating the expression of toxin and virulence factors. This study aimed to identify reliable reference genes for normalizing reverse transcription quantitative PCR (RT-qPCR) data in toxigenic B. cereus. An emetic and an enteropathogenic strain were used as model organisms to establish a suitable reference gene set to monitor the dynamics of toxin gene transcription. Ten candidate reference genes were evaluated for their expression stability using geNorm, NormFinder, BestKeeper and the ΔCq method, with the final rankings integrated via RefFinder. Among the tested genes, rho, rpoD and recA were identified as the most stable expressed reference genes across all tested conditions. As shown in this proof-of-principle study, the established reference gene set provides a suitable tool to investigate the influence of extrinsic and intrinsic factors on toxin gene transcription. In conclusion, our newly established reference gene set provides a robust basis for studying toxin gene expression in B. cereus and contributes to a better understanding of its pathogenicity and potential strategies to mitigate its harmful effects.

The transcriptional analysis of pepper shed light on a proviral role of light-harvesting chlorophyll a/b binding protein 13 during infection of pepper mild mottle virus

Pepper mild mottle virus (PMMoV), a member of the genus Tobamovirus, causes severe damage on pepper worldwide. Despite its impact, the pathogenicity mechanisms of PMMoV and the pepper plant’s response to infection remain poorly understood. Here, we compared the transcriptomic changes in a susceptible pepper inbred line 21C241 with a resistant inbred line 21C385 seedlings, following systemic PMMoV infection using RNA sequencing. Our results revealed that PMMoV induced more pronounced mosaic symptoms and higher viral accumulation levels in the susceptible line 21C241 compared to the resistant line 21C385. We identified 462 and 401 differentially expressed genes (DEGs) in the systemically-infected leaves of the susceptible and resistant lines, respectively, when compared to their healthy counterparts. The majority of these DEGs were involved in photosynthesis and the biosynthesis of secondary metabolites, with 28 DEGs exhibiting distinct expression patterns between the two lines. Notably, the expression level of the chlorophyll a-b binding protein 13 (CAB13) was significantly up-regulated in resistant line 21C385 following PMMoV infection. Functional analysis through silencing of CAB13 in pepper and Nicotiana benthamiana demonstrated a reduction in PMMoV accumulation, suggesting that CAB13 plays a positive role in facilitating PMMoV infection in pepper plants. Taken together, our findings highlight the distinct gene expression profiles between susceptible and resistant pepper lines in response to PMMoV infection and confirm the proviral role of CAB13. This study provides valuable insights into the molecular mechanisms underlying resistance and susceptibility in pepper plants and may inform future strategies for disease management.