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Background Past research has shown that virus-induced phytoene desaturase ( PDS ) gene silencing via agroinjection in the attached and detached fruit of tomato plants results in a pale-yellow fruit phenotype. Although the PDS gene is often used as a marker for gene silencing in tomatoes, little is known about the role of PDS in fruit ripening. In this study, we investigated whether the pepper PDS gene silenced endogenous PDS genes in the fruit of two tomato cultivars, Dotaerang Plus and Legend Summer. Results We found that the pepper PDS gene successfully silenced endogenous PDS in tomato fruit at a silencing frequency of 100% for both cultivars. A pale-yellow silenced area was observed over virtually the entire surface of individual fruit due to the transcriptional reduction in phytoene desaturase ( PDS ), zeta-carotene ( ZDS ), prolycopene isomerase ( CrtlSO ), and beta-carotene hydroxylase ( CrtR - b2 ), which are the carotenoid biosynthesis genes responsible for the red coloration in tomatoes. PDS silencing also affected the expression levels of the fruit-ripening genes Tomato AGAMOUS-LIKE1 ( TAGL1 ), RIPENING INHIBITOR ( RIN ), pectin esterase gene ( PE ), lipoxygenase ( LOX ), FRUITFULL1/FRUITFUL2 ( FUL1/FUL2 ), and the ethylene biosynthesis and response genes 1-aminocyclopropane-1-carboxylate oxidase 1 and 3 ( ACO1 and ACO3 ) and ethylene-responsive genes ( E4 and E8 ). Conclusion These results suggest that PDS is a positive regulator of ripening in tomato fruit, which must be considered when using it as a marker for virus-induced gene silencing (VIGS) experiments in order to avoid fruit-ripening side effects.

A maternal mortality ratio is a sensitive indicator when comparing the overall maternal health between countries and its very high figure indicates the failure of maternal healthcare efforts. Cambodia, Laos, Myanmar, and Vietnam-CLMV countries are the low-income countries of the South-East Asia region where their maternal mortality ratios are disproportionately high. This systematic review aimed to summarize all possible factors influencing maternal mortality in CLMV countries. This systematic review applied \"The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) Checklist (2020)\", Three key phrases: \"Maternal Mortality and Health Outcome\", \"Maternal Healthcare Interventions\" and \"CLMV Countries\" were used for the literature search. 75 full-text papers were systematically selected from three databases (PubMed, Google Scholar and Hinari). Two stages of data analysis were descriptive analysis of the general information of the included papers and qualitative analysis of key findings. Poor family income, illiteracy, low education levels, living in poor households, and agricultural and unskilled manual job types of mothers contributed to insufficient antenatal care. Maternal factors like non-marital status and sex-associated work were highly associated with induced abortions while being rural women, ethnic minorities, poor maternal knowledge and attitudes, certain social and cultural beliefs and husbands' influences directly contributed to the limitations of maternal healthcare services. Maternal factors that made more contributions to poor maternal healthcare outcomes included lower quintiles of wealth index, maternal smoking and drinking behaviours, early and elderly age at marriage, over 35 years pregnancies, unfavourable birth history, gender-based violence experiences, multigravida and higher parity. Higher unmet needs and lower demands for maternal healthcare services occurred among women living far from healthcare facilities. Regarding the maternal healthcare workforce, the quality and number of healthcare providers, the development of healthcare infrastructures and human resource management policy appeared to be arguable. Concerning maternal healthcare service use, the provisions of mobile and outreach maternal healthcare services were inconvenient and limited. Low utilization rates were due to several supply-side constraints. The results will advance knowledge about maternal healthcare and mortality and provide a valuable summary to policymakers for developing policies and strategies promoting high-quality maternal healthcare.

The β1 integrin subunit contributes to pancreatic beta cell growth and function through communication with the extracellular matrix (ECM). The effects of in vitro and in vivo β1 integrin knockout have been extensively studied in mature islets, yet no study to date has examined how the loss of β1 integrin during specific stages of pancreatic development impacts beta cell maturation. Beta-cell-specific tamoxifen-inducible Cre recombinase (MIP-CreERT) mice were crossed with mice containing floxed Itgb1 (β1 integrin) to create an inducible mouse model (MIPβ1KO) at the second transition stage (e13.5) of pancreas development. By e19.5–20.5, the expression of beta-cell β1 integrin in fetal MIPβ1KO mice was significantly reduced and these mice displayed decreased beta cell mass, density and proliferation. Morphologically, fetal MIPβ1KO pancreata exhibited reduced islet vascularization and nascent endocrine cells in the ductal region. In addition, decreased ERK phosphorylation was observed in fetal MIPβ1KO pancreata. The expression of transcription factors needed for beta-cell development was unchanged in fetal MIPβ1KO pancreata. The findings from this study demonstrate that β1 integrin signaling is required during a transition-specific window in the developing beta-cell to maintain islet mass and vascularization.

Schizophrenia (SZ) is a severe, complex, and common mental disorder with high heritability (80%), an adult age of onset, and high discordance (∼50%) in monozygotic twins (MZ). Extensive studies on familial and non-familial cases have implicated a number of segregating mutations and de novo changes in SZ that may include changes to the mitochondrial genome. Yet, no single universally causal variant has been identified, highlighting its extensive genetic heterogeneity. This report specifically focuses on the assessment of changes in the mitochondrial genome in a unique set of monozygotic twins discordant (MZD) for SZ using blood. Genomic DNA from six pairs of MZD twins and two sets of parents (N = 16) was hybridized to the Affymetrix Human SNP Array 6.0 to assess mitochondrial DNA copy number (mtDNA-CN). Whole genome sequencing (WGS) and quantitative polymerase chain reaction (qPCR) was performed for a subset of MZD pairs and their parents and was also used to derive mtDNA-CN estimates. The WGS data were further analyzed to generate heteroplasmy (HP) estimates. Our results show that mtDNA-CN estimates for within-pair and mother-child differences were smaller than comparisons involving unrelated individuals, as expected. MZD twins showed discordance in mtDNA-CN estimates and displayed concordance in directionality of differences for mtDNA-CN across all technologies. Further, qPCR performed better than Affymetrix in estimating mtDNA-CN based on relatedness. No reliable differences in HP were detected between MZD twins. The within-MZD differences in mtDNA-CN observed represent postzygotic somatic changes that may contribute to discordance of MZ twins for diseases, including SZ.

Background According to anecdotal reports, SDF’s ability to arrest caries can be enhanced by light-curing in a clinical setting. The purpose of the present study was to explore the dental professionals’ perceptions of using SDF and to understand the barriers and enabling factors to using SDF with and without light-curing. Methods A qualitative study was conducted with dental professionals who had experience with using SDF with and without light-curing. A purposive heterogeneous and convenience sampling approach was applied to ensure the inclusion of participants with different employment and experience levels. Eighteen participants (dental students (undergraduate and postgraduate), dental personnel (practicing dentists and dental hygienists), and academics) with ages ranging from 22 to 53 years old participated in the study. The data were collected through a semi-structured open-ended questionnaire. Manual line-by-line coding and content analysis methods were used for data analysis. Results Most participants indicated a preference for light-curing, citing perceived benefits, such as quicker treatment, convenience, better visibility, prevention of saliva contamination, and ensuring thorough SDF coverage. However, some participants voiced concerns, based either on existing practice guidelines or due to insufficient evidence regarding the efficacy of light-curing. Additionally, one participant proposed a hybrid approach based on tooth location, suggesting avoiding using light-curing on anterior teeth, while utilizing it on posterior teeth. Conclusion The participants’ responses indicated that the use of SDF with and without light-curing each has its merits. Using SDF with light-curing emerged as the preferred method due to perceived benefits. e.g., quicker treatment, better moisture control. However, concerns regarding existing guidelines and the lack of robust evidence for its efficacy were also identified. This study highlights the need for additional research to address the knowledge gaps and provide stronger evidence to support the use of light-curing after SDF application.

The unfolded protein response (UPR) is activated in pancreatic pathologies and suggested as a target for therapeutic intervention. In this study, we examined activating transcription factor 3 (ATF3), a mediator of the UPR that promotes acinar-to-ductal metaplasia (ADM) in response to pancreatic injury. Since ADM is an initial step in the progression to pancreatic ductal adenocarcinoma (PDAC), we hypothesized that ATF3 is required for initiation and progression of PDAC. We generated mice carrying a germline mutation of Atf3 ( Atf3 −/− ) combined with acinar-specific induction of oncogenic KRAS ( Ptf1a creERT/+ Kras G12D/+ ). Atf3 −/− mice with (termed APK ) and without KRAS G12D were exposed to cerulein-induced pancreatitis. In response to recurrent pancreatitis, Atf3 −/− mice showed decreased ADM and enhanced regeneration based on morphological and biochemical analysis. Similarly, an absence of ATF3 reduced spontaneous pancreatic intraepithelial neoplasia (PanIN) formation and PDAC in Ptf1a creERT/+ Kras G12D/+ mice. In response to injury, KRAS G12D bypassed the requirement for ATF3 with a dramatic loss in acinar tissue and PanIN formation observed regardless of ATF3 status. Compared to Ptf1a creERT/+ Kras G12D/+ mice, APK mice exhibited a significant decrease in pancreatic and total body weight, did not progress through to PDAC, and showed altered pancreatic fibrosis and immune cell infiltration. These findings suggest a complex, multifaceted role for ATF3 in pancreatic cancer pathology.

Objective Mitochondrial dysfunction and nuclear epigenetic alterations, two hallmarks of aging, are associated with aberrant development and complex disease risk. Here, we report a method for the simultaneous assessment of mitochondrial DNA copy number (mtDNA-CN) and DNA methylation age (DNAm age) from the same DNA extraction using quantitative polymerase chain reaction (qPCR) and array data, respectively. Result We present methods for the concurrent estimation of mtDNA-CN and DNAm age from the same DNA samples. This includes qPCR to estimate mtDNA-CN, representing the number of circular mitochondrial genomes in a cell, and DNA methylation microarray data to estimate the epigenetic age of an individual. Further, we provide a method for the combination of these metrics into a shared metric termed ‘mtEpiAge’. This approach provides a valuable tool for exploring the interplay between mitochondrial dysfunction and nuclear epigenetic alterations, and their associations with disease and aging.

In this study, whether the addition of antifreeze protein (AFP) to a cryopreservative solution (plant vitrification solution 2 (PVS2)) is more effective in reducing freezing injuries in Hosta capitata than PVS2 alone at different cold exposure times (6, 24, and 48 h) is investigated. The upregulation of C-repeat binding factor 1 (CBF1) and dehydrin 1 (DHN1) in response to low temperature was observed in shoots. Shoots treated with distilled water (dH2O) strongly triggered gene expression 6 h after cold exposure, which was higher than those expressed in PVS2 and PVS2+AFP. However, 24 h after cold exposure, gene expressions detected in dH2O and PVS2 treatments were similar and higher than PVS2 + AFP. The expression was highest in PVS2+AFP when the exposure time was extended to 48 h. Similarly, nitric reductase activities 1 and 2 (Nia1 and Nia2) genes, which are responsible for nitric oxide production, were also upregulated in low-temperature-treated shoots, as observed for CBF1 and DHN1 expression patterns during cold exposure periods. Based on the gene expression patterns, shoots treated with PVS2+AFP were more likely to resist cold stress, which was also associated with the higher cryopreservation efficiency of PVS2+AFP compared to PVS2 alone. This finding suggests that the improvement of cryopreservation efficiency by AFP could be due to the transcriptional regulation of CBF1, DHN1, Nia1, and Nia2, which might reduce freezing injuries during cryopreservation. Thus, AFP could be potentially used as a cryoprotectant in the cryopreservation of rare and commercially important plant germplasm.

Circular RNAs (circRNAs) participate in complex diseases and have emerged as a novel biomarker and therapeutic target. It is challenging to identify circRNAs from total RNA-seq data and to validate and quantify their levels by qRT-PCR. We sought to characterize the circRNA transcriptome from RNA-seq with total RNA from septic peripheral blood mononuclear cells (PBMCs), assess/compare commonly used aligners, and optimize validation methods, considering sepsis is the leading cause of death in intensive care units (ICU) and there is no effective treatment. PBMCs were isolated from sepsis patients before and after intensive care, and RNA sequencing was conducted with total RNA. RNA-seq data for circRNAs were analyzed using four pipelines. A circRNA transcriptome has been generated, and four alignment and annotation pipelines have been evaluated. TopHat was found to be the most sensitive aligner while MapSplice was found to be the most accurate. Overall, circRNA expression in septic PBMCs were found to be more abundant and less diverse at ICU admission (ICU-AD) compared with ICU discharge (ICU-DC). There were three differentially expressed circRNAs between ICU-AD and ICU-DC. The addition of reverse primers to a reverse transcription reaction improved reproducibility and accuracy of qRT-PCR. Among these altered circRNAs, two different isoforms of circular RNA from the ASPH gene (circASPH(2,4), and circASPH(2,3,4) with the same backsplicing junction (BSJ) were identified and specifically quantified by qRT-PCR. All the two isoforms were highly expressed at ICU-AD and circASPH(2,3,4) levels positively correlated with the length of stay in ICU. The circRNAs predictably bind to miR-670, miR-7975, miR-6818, and miR-384 and interact with proteins TLR2 and ZC3H12D. In conclusion, it is feasible to identify circRNA transcriptome from RNA-seq with total RNA. Moreover, RT-PCR followed by gel electrophoresis is important to identify/ distinguish a new isoform of a circRNA with the same BSJ. CircRNA transcriptome in septic PBMCs evolves as infectious severity changes. CircASPH(2,3,4) has a clinical potential as an indicator of disease severity.