Different Effects of Total Bilirubin on 90-Day Mortality in Hospitalized Patients With Cirrhosis and Advanced Fibrosis: A Quantitative Analysis

Introduction: Total bilirubin (TB) is a major prognosis predictor representing liver failure in patients with acute on chronic liver failure (ACLF). However, the cutoff value of TB for liver failure and whether the same cutoff could be applied in both cirrhotic and non-cirrhotic patients remain controversial. There is a need to obtain the quantitative correlation between TB and short-term mortality via evidence-based methods, which is critical in establishing solid ACLF diagnostic criteria.Methods: Patients hospitalized with cirrhosis or advanced fibrosis (FIB-4 > 1.45) were studied. TB and other variables were measured at baseline. The primary outcome was 90-day transplantation-free mortality. Multi-variable Cox proportional hazard model was used to present the independent risk of mortality due to TB. Generalized additive model and second derivate (acceleration) were used to plot the “TB-mortality correlation curves.” The mathematical (maximum acceleration) and clinical (adjusted 28-day transplantation-free mortality rate reaching 15%) TB cutoffs for liver failure were both calculated.Results: Among the 3,532 included patients, the number of patients with cirrhosis and advanced fibrosis were 2,592 and 940, respectively, of which cumulative 90-day mortality were 16.6% (430/2592) and 7.4% (70/940), respectively. Any increase of TB was found the independent risk factor of mortality in cirrhotic patients, while only TB >12 mg/dL independently increased the risk of mortality in patients with advanced fibrosis. In cirrhotic patients, the mathematical TB cutoff for liver failure is 14.2 mg/dL, with 23.3% (605/2592) patients exceeding it, corresponding to 13.3 and 25.0% adjusted 28- and 90-day mortality rate, respectively. The clinical TB cutoff for is 18.1 mg/dL, with 18.2% (471/2592) patients exceeding it. In patients with advanced fibrosis, the mathematical TB cutoff is 12.1 mg/dL, 33.1% (311/940) patients exceeding it, corresponding to 2.9 and 8.0% adjusted 28- and 90-day mortality rate, respectively; the clinical TB cutoff was 36.0 mg/dL, 1.3% (12/940) patients above it.Conclusion: This study clearly demonstrated the significantly different impact of TB on 90-day mortality in patients with cirrhosis and advanced fibrosis, proving that liver failure can be determined by TB alone in cirrhosis but not in advanced fibrosis. The proposed TB cutoffs for liver failure provides solid support for the establishment of ACLF diagnostic criteria

Analysis of pharmacogenetic interations between bmp signaling and wdr68 and wdr68 activity in zebrafish craniofacial development

Bone morphogenetic protein (Bmp) signaling is required for neural crest cell induction, proliferation, migration, and patterning. In zebrafish, the neural crest cells within the first pharyngeal arch develop into both the Meckel???s cartilage (M) and palatoquadrate (PQ). Dorsomorphin (DM) and isoliquiritigenin (ISL) are known to repress and activate the Bmp pathway, respectively. I hypothesized that Wdr68 modulates Bmp signaling. I saw synergistic reductions of M and PQ cartilages in DM-treated wdr68hi3812/hi3812 mutant zebrafish embryos relative to DM-treated wild-type siblings. I saw rescue of M cartilage in ISL-treated wdr68hi3812/hi3812 mutant zebrafish embryos relative to mutant siblings. Bmp is known to induce endothelin-1 (edn1) expression, which is known to be important for M cartilage formation. I saw further reduction in edn1 expression in wdr68hi3812/hi3812 mutant zebrafish embryos treated with DM compared to expression in mutant zebrafish embryos alone. I also saw rescued edn1 expression in wdr68hi3812/hi3812 mutant zebrafish embryos treated with ISL compared to expression in mutant zebrafish embryos alone. Thus Wdr68 is important for robust Bmp signaling to downstream edn1 gene expression

Combined Transcriptomic and Metabolomic Analysis Reveals the Mechanism of Flavonoid Biosynthesis in <i>Handroanthus chrysanthus</i> (Jacq.) S.O.Grose

Handroanthus and Tabebuia are known for their ornamental and medicinal value, which are attributed to metabolites. However, the mechanisms underlying the synthesis of these metabolites are poorly understood. In this study, the expression levels of secondary metabolites and the mechanism of flavonoid biosynthesis in the bark and leaves of Handroanthus chrysantha (Jaq.) were examined using transcriptomic and metabolomic techniques. Metabolic analysis identified several differentially accumulated metabolites (DAMs), most of which were flavonoids, isoprenoids, and sterols. Additionally, 30 flavonoids were identified in the bark and leaves of H. chrysantha. Transcriptomic analysis identified 69 genes involved in flavonoid biosynthesis, among which 49 were significantly different between the bark and leaves. qRT-PCR analysis of eight genes involved in flavonoid biosynthesis showed that the expression patterns of the genes were consistent with the transcriptome sequencing data. Integrative transcriptomic and metabolomic analysis showed that 20 differentially expressed genes (DEGs) associated with flavonoid biosynthesis were strongly correlated with seven DAMs, confirming the involvement of the DEGs in flavonoid biosynthesis. These findings considerably contribute to the understanding of the biosynthesis of secondary metabolites in H. chrysantha and serve as a reference for further pharmacological studies

Overexpression of <i>PvFAD3</i> Gene from <i>Plukenetia volubilis</i> Promotes the Biosynthesis of α-Linolenic Acid in Transgenic Tobacco Seeds

The ω-3 fatty acid desaturase (FAD3) gene encodes a rate-limiting enzyme in the synthesis of α-linolenic acid. In this study, homologous cloning was used to obtain the full-length sequence of the PvFAD3 gene of Plukenetia volubilis. The full-length DNA sequence was 1871 bp long, with 8 exons and 7 introns. The structural analysis of the amino acid sequence revealed that the PvFAD3 protein contained three histidine-conserved regions and an endoplasmic reticulum retention signal. The real-time reverse transcription-polymerase chain reaction performed for determining the expression patterns of the PvFAD3 gene in different tissues of P. volubilis showed that PvFAD3 expression was highly expressed in the fast oil accumulation stage of seed. The analysis of subcellular localization assay in epidermal cells of tobacco (Nicotiana benthamiana) leaves showed that the PvFAD3 protein was mainly localized in the endoplasmic reticulum. Seed-specific overexpression vectors were constructed, and Agrobacterium-mediated genetic transformation was performed to obtain transgenic tobacco plants overexpressing PvFAD3. The results of fatty acid assays performed using harvested seeds showed a significant increase in α-linolenic acid content, a dramatic decrease in linoleic acid content, and an obvious increase in oil content in transgenic tobacco seeds. Collectively, the PvFAD3 gene of P. volubilis was confirmed as a key enzyme gene for α-linolenic acid synthesis; thus, indicating that the PvFAD3 gene can be used for fatty acid fraction improvement in oilseed plants

Integrated Transcriptome and Proteome Analysis Provides Insight into the Ribosome Inactivating Proteins in Plukenetia volubilis Seeds

Plukenetia volubilis is a highly promising plant with high nutritional and economic values. In our previous studies, the expression levels of ricin encoded transcripts were the highest in the maturation stage of P. volubilis seeds. The present study investigated the transcriptome and proteome profiles of seeds at two developmental stages (Pv-1 and Pv-2) using RNA-Seq and iTRAQ technologies. A total of 53,224 unigenes and 6026 proteins were identified, with functional enrichment analyses, including GO, KEGG, and KOG annotations. At two development stages of P. volubilis seeds, 8815 unique differentially expressed genes (DEGs) and 4983 unique differentially abundant proteins (DAPs) were identified. Omics-based association analysis showed that ribosome-inactivating protein (RIP) transcripts had the highest expression and abundance levels in Pv-2, and those DEGs/DAPs of RIPs in the GO category were involved in hydrolase activity. Furthermore, 21 RIP genes and their corresponding amino acid sequences were obtained from libraries produced with transcriptome analysis. The analysis of physicochemical properties showed that 21 RIPs of P. volubilis contained ricin, the ricin_B_lectin domain, or RIP domains and could be divided into three subfamilies, with the largest number for type II RIPs. The expression patterns of 10 RIP genes indicated that they were mostly highly expressed in Pv-2 and 4 transcripts encoding ricin_B_like lectins had very low expression levels during the seed development of P. volubilis. This finding would represent valuable evidence for the safety of oil production from P. volubilis for human consumption. It is also notable that the expression level of the Unigene0030485 encoding type I RIP was the highest in roots, which would be related to the antiviral activity of RIPs. This study provides a comprehensive analysis of the physicochemical properties and expression patterns of RIPs in different organs of P. volubilis and lays a theoretical foundation for further research and utilization of RIPs in P. volubilis

SLViT: Shuffle-convolution-based lightweight Vision transformer for effective diagnosis of sugarcane leaf diseases

Farmers must accurately and promptly identify sugarcane leaf diseases with identical symptoms. RGB images have a beneficial function in disease identification. Nevertheless, complex backgrounds and identical symptoms can significantly reduce the recognition accuracy and robustness. To overcome these challenges, the SLViT hybrid network is presented, in which the transformer encoder is converted to a flexible plug-in (LViT) that is subsequently integrated into several locations of a lightweight CNN architecture (SHDC). SLViT is initially trained on the publicly available disease dataset Plant Village before being moved to the self-created sugarcane leaf disease dataset SLD10k, which consists of seven classes and 10,309 images. The ablation experiments demonstrate that all the adjustments to SLViT have contributed positively to its overall performance. SLViT outperforms six SOTA models and three custom-designed leaf-disease recognition models on Plant Village in terms of speed (1,832 FPS), weight (2 MB), consumption (50 M), and precision (98.84 %). SLViT also outperformed MobileNetV3_small on the SLD10k dataset with an accuracy bonus of 1.87 % and a size reduction of 66.3 %. The experiment also reveals that SLViT has absorbed the advantages of both the lightweight CNN and the noise-resistant transformer. This study demonstrates the applicability of SLViT for sugarcane leaf diagnosis in the field

Expression Patterns and Regulation of Non-Coding RNAs during Synthesis of Cellulose in Eucalyptus grandis Hill

Cellulose, an essential structural component in the plant cell wall and a renewable biomass resource, plays a significant role in nature. Eucalyptus’s excellent timber tree species (including Eucalyptus grandis Hill) provide many raw materials for the paper and wood industries. The synthesis of cellulose is a very complex process involving multiple genes and regulated by various biological networks. However, research on regulating associated genes and non-coding RNAs during cellulose synthesis in E. grandis remains lacking. In this study, the wood anatomical characteristics and chemical indexes of E. grandis were analyzed by taking three different parts (diameter at breast height (DBH), middle and upper part of the trunk) from the main stem of E. grandis as raw materials. The role of non-coding RNAs (Long non-coding RNA, lncRNA; Micro RNA, miRNA; Circle RNA, circRNA) on regulating candidate genes was presented, and the network map of ceRNA (Competing endogenous RNA) regulation during wood cellulose biosynthesis of E. grandis was constructed. The transcriptome sequencing of nine samples obtained from the trunk of the immature xylem in E. grandis at DBH, middle and upper parts had a 95.81 G clean reading, 57,480 transcripts, 7365 lncRNAs, and 5180 circRNAs. Each sample had 172–306 known miRNAs and 1644–3508 new miRNAs. A total of 190 DE-lncRNAs (Differentially expressed long non-coding RNAs), 174 DE-miRNAs (Differentially expressed micro RNAs), and 270 DE-circRNAs (Differentially expressed circle RNAs) were obtained by comparing transcript expression levels. Four lncRNAs and nine miRNAs were screened out, and the ceRNA regulatory network was constructed. LncRNA1 and lncRNA4 regulated the genes responsible for cellulose synthesis in E. grandis, which were overexpressed in 84K (Populus Alba × Populus glandulosa) poplar. The cellulose and lignin content in lncRNA4-oe were significantly higher than wild type 84K poplar and lncRNA1-oe. The average plant height, middle and basal part of the stem diameter in lncRNA4-oe were significantly higher than the wild type. However, there was no significant difference between the growth of lncRNA1-oe and the wild type. Further studies are warranted to explore the molecular regulatory mechanism of cellulose biosynthesis in Eucalyptus species

Comparative transcriptomics and bioinformatics analysis of genes related to photosynthesis in Eucalyptus camaldulensis

The timber species Eucalyptus camaldulensis is one of the most important in southern China. Therefore, it is essential to understand the photosynthetic pattern in eucalyptus leaves. In the present study, eighteen photosynthesis-related genes were analyzed using bioinformatics methods. The results indicated that there were ten differentially expressed ribose-5-phosphate isomerase genes (RPI), and six of them were up-regulated in the mature leaves compared to the young leaves, while others were down-regulated. The differential expression of four rubisco methyltransferase genes (RBCMT) were observed. Two of them were up-regulated, while two were down-regulated in mature leaves compared to young leaves. Furthermore, two ribulose-phosphate-3-epimerase genes (RPE) were up-regulated in the mature leaves compared to the young leaves. In contrast, two genes involved in triosephosphate isomerase (TIM) were down-regulated in mature leaves compared with young leaves. The current study provides basic information about the transcriptome of E. camaldulensis and lays a foundation for further research in developing and utilizing important photosynthetic genes

Combining Quantitative Data on Growth, Wood Density and Other Traits with SSR Markers to Evaluate Genetic Diversity and Structure in a Planted Population of Eucalyptus camaldulensis Dehn.

Eucalyptus camaldulensis Dehn. is one of the most morphologically and genetically variable Eucalyptus species. Growth, Leptocybe invasa Fisher &amp; La Salle susceptibility, pilodyn penetration and other traits up to age 36 months were assessed in a seed source/family trial in China comprising 112 seedlots representing five natural stand and six exotic seed sources. Genetic diversity and population structure of this trial population were also analyzed using 48 simple sequence repeat (SSR) markers. The key objective was to examine whether the genomic data could provide value over information obtained from just quantitative trait data. Significant genetic variation was found among seed sources and among families within seed sources for most quantitative traits. The ratio of variance among seed sources to variance among families within seed sources, based on variances estimated from quantitative trait data, varied from 0.1% (height at 9 months) up to 75.2% (bark thickness). Equivalent ratios estimated from the AMOVA on SSR loci data were similar for height (ages 24 and 36 months) and also pilodyn penetration at 36 months, but not for 9-month height or 36-month bark thickness. From 48 SSR loci examined, the genetic differentiation coefficient (among seed sources) was 0.086, indicating low genetic differentiation among seed sources. While overall genetic diversity in the trial population examined was high, the levels within the different seed sources varied markedly. Prior to this study, genetic distances among families from the three exotic seed sources (from domesticated Indian populations) in the trial, along with their genetic distances from, and relatedness to, families from five natural stand seed sources (Australian) in the trial were unknown. The SSR loci data removed uncertainties and revealed that the exotic sources increased the breadth of genetic origins represented in the trial population&mdash;information that could not have been obtained from just the quantitative trait data

Preoxygenation with standard facemask combining apnoeic oxygenation using high flow nasal cannula versuss standard facemask alone in patients with and without obesity: the OPTIMASK international study

Abstract Background Combining oxygen facemask with apnoeic oxygenation using high-flow-nasal-oxygen (HFNO) for preoxygenation in the operating room has not been studied against standard oxygen facemask alone. We hypothesized that facemask-alone would be associated with lower levels of lowest end-tidal oxygen (EtO2) within 2 min after intubation in comparison with facemask combined with HFNO. Methods In an international prospective before–after multicentre study, we included adult patients intubated in the operating room from September 2022 to December 2022. In the before period, preoxygenation was performed with facemask-alone, which was removed during laryngoscopy. In the after period, facemask combined with HFNO was used for preoxygenation and HFNO for apnoeic oxygenation during laryngoscopy. HFNO was maintained throughout intubation. The primary outcome was the lowest EtO2 within 2 min after intubation. The secondary outcome was SpO2 ≤ 95% within 2 min after intubation. Subgroup analyses were performed in patients without and with obesity. This study was registered 10 August 2022 with ClinicalTrials.gov, number NCT05495841. Results A total of 450 intubations were evaluated, 233 with facemask-alone and 217 with facemask combined with HFNO. In all patients, the lowest EtO2 within 2 min after intubation was significantly lower with facemask-alone than with facemask combined with HFNO, 89 (85–92)% vs 91 (88–93)%, respectively (mean difference − 2.20(− 3.21 to − 1.18), p < 0.001). In patients with obesity, similar results were found [87(82–91)% vs 90(88–92)%, p = 0.004]; as in patients without obesity [90(86–92)% vs 91(89–93)%, p = 0.001)]. SpO2 ≤ 95% was more frequent with facemask-alone (14/232, 6%) than with facemask combined with HFNO (2/215, 1%, p = 0.004). No severe adverse events were recorded. Conclusions Combining facemask with HFNO for preoxygenation and apnoeic oxygenation was associated with increased levels of lowest EtO2 within 2 min after intubation and less desaturation