Association between clinical features and decreased degree centrality and variability in dynamic functional connectivity in the obsessive-compulsive disorder.

Neuroimaging studies have indicated widespread brain structural and functional disruptions in patients with obsessive-compulsive disorder (OCD). However, the underlying mechanism of these changes remains unclear. A total of 45 patients with OCD and 42 healthy controls (HC) were enrolled. The study investigated local degree centrality (DC) abnormalities and employed abnormal regions of DC as seeds to investigate variability in dynamic functional connectivity (dFC) in the whole brain using a sliding window approach to analyze resting-state functional magnetic resonance imaging. The relationship between abnormal DC and dFC as well as the clinical features of OCD were examined using correlation analysis. Our findings suggested decreased DC in the bilateral thalamus, bilateral precuneus, and bilateral cuneus in OCD patients and a nominally negative correlation between the DC value in the thalamus and illness severity measured using the Yale-Brown Obsessive Compulsive Scale (Y-BOCS). In addition, seed-based dFC analysis showed that compared to measurements in the HC, the patients had decreased dFC variability between the left thalamus and the left cuneus and right lingual gyrus, and between the bilateral cuneus and bilateral postcentral gyrus, and a nominally positive correlation between the duration of illness and dFC variability between the left cuneus and left postcentral gyrus. These results indicated that OCD patients had decreased hub importance in the bilateral thalamus and cuneus throughout the entire brain. This reduction was associated with impaired coupling with dynamic function in the visual cortex and sensorimotor network and provided novel insights into the neurophysiological mechanisms underlying OCD.

The impact of deep learning image reconstruction of spectral CTU virtual non contrast images for patients with renal stones.

Purpose: To compare image quality and detection accuracy of renal stones between deep learning image reconstruction (DLIR) and Adaptive Statistical Iterative Reconstruction-Veo (ASIR-V) reconstructed virtual non-contrast (VNC) images and true non-contrast (TNC) images in spectral CT Urography (CTU). Methods: A retrospective analysis was conducted on images of 70 patients who underwent abdominal-pelvic CTU in TNC phase using non-contrast scan and contrast-enhanced corticomedullary phase (CP) and excretory phase (EP) using spectral scan. The TNC scan was reconstructed using ASIR-V70 % (TNC-AR70), contrast-enhanced scans were reconstructed using AR70, DLIR medium-level (DM), and high-level (DH) to obtain CP-VNC-AR70/DM/DH and EP-VNC-AR70/DM/DH image groups, respectively. CT value, image quality and kidney stones quantification accuracy were measured and compared among groups. The subjective evaluation was independently assessed by two senior radiologists using the 5-point Likert scale for image quality and lesion visibility. Results: DH images were superior to AR70 and DM images in objective image quality evaluation. There was no statistical difference in the liver and spleen (both P > 0.05), or within 6HU in renal and fat in CT value between VNC and TNC images. EP-VNC-DH had the lowest image noise, highest SNR, and CNR, and VNC-AR70 images had better noise and SNR performance than TNC-AR70 images (all p < 0.05). EP-VNC-DH had the highest subjective image quality, and CP-VNC-DH performed the best in lesion visibility. In stone CT value and volume measurements, there was no statistical difference between VNC and TNC (P > 0.05). Conclusion: The DLIR-reconstructed VNC images in CTU provide better image quality than the ASIR-V reconstructed TNC images and similar quantification accuracy for kidney stones for potential dose savings.The study highlights that deep learning image reconstruction (DLIR)-reconstructed virtual non-contrast (VNC) images in spectral CT Urography (CTU) offer improved image quality compared to traditional true non-contrast (TNC) images, while maintaining similar accuracy in kidney stone detection, suggesting potential dose savings in clinical practice.

Fermentation broth of a novel endophytic fungus enhanced maize salt tolerance by regulating sugar metabolism and phytohormone biosynthesis or signaling.

Soil salinization is a major environmental factor that severely affects global agriculture. Root endophytes can enter root cells, and offer various ecological benefits, such as promoting plant growth, improving soil conditions, and enhancing plant resistance. Su100 is a novel strain of endophytic fungus that was characterized from blueberry roots. In this study, we focused on evaluating the effects of Su100 secretion on maize growth. The results demonstrated that maize treated with Su100 fermentation broth (SFB) exhibited significantly stronger salt tolerance than the control. It is worth mentioning that the treated root system not only had an advantage in terms of biomass but also a change in root structure with a significant increase in lateral roots (LRs) compared to the control. Transcriptome analysis combined with hormone content measurements indicated that SFB upregulated the auxin signaling pathway, and also caused alterations in brassinosteroids (BR) and jasmonic acid (JA) biosynthesis and signaling pathways. Transcriptome analyses also indicated that SFB caused significant changes in the sugar metabolism of maize roots. The major changes included: enhancing the conversion and utilization of sucrose in roots; increasing carbon flow to uridine diphosphate glucose (UDPG), which acted as a precursor for producing more cell wall polysaccharides, mainly pectin and lignin; accelerating the tricarboxylic acid cycle, which were further supported by sugar content determinations. Taken together, our results indicated that the enhanced salt tolerance of maize treated with SFB was due to the modulation of sugar metabolism and phytohormone biosynthesis or signaling pathways. This study provided new insights into the mechanisms of action of endophytic fungi and highlighted the potential application of fungal preparations in agriculture.

A novel growth-promoting dark septate endophytic fungus improved drought tolerance in blueberries by modulating phytohormones and non-structural carbohydrates.

Drought is a significant global issue affecting agricultural production, and the utilization of beneficial rhizosphere microorganisms is one of the effective ways to increase the productivity of crops and forest under drought. In this study, we characterized a novel growth-promoting dark septate endophytes (DSE) fungus R16 (Dothideomycetes sp.) derived from blueberry roots. Hyphae or microsclerotia were visible within the epidermal or cortical cells of R16-colonized blueberry roots, which was consistent with the typical characteristics of DSE fungi. Inoculation with R16 promoted the growth of blueberry seedlings, and the advantage over the control group was more significant under PEG-induced drought. Comparison of physiological indicators related to drought resistance between the inoculated and control groups was performed on the potted blueberry plants, including the chlorophyll content, net photosynthetic rate, root activities, malondialdehyde and H2O2 content, which indicated that R16 colonization mitigated drought injury in blueberry plants. We further analyzed the effects of R16 on phytohormones and non-structural carbohydrates (NSCs) to explore the mechanism of increased drought tolerance by R16 in blueberry seedlings. The results showed that except for the gibberellin content, indole-3-acetic acid, zeatin and abscisic acid varied significantly between the inoculated and control groups. Sucrose phosphate synthase and sorbitol-6-phosphate dehydrogenase activities in mature leaves, the key enzymes responsible for sucrose and sorbitol synthesis, respectively, as well as sorbitol dehydrogenase, sucrose synthase, cell wall invertase, hexokinase and fructokinase in roots, the key enzymes involved in the NSCs metabolism, showed significant differences between the inoculated and control groups before and after drought treatment. These results suggested that the positive effects of R16 colonization on the drought tolerance of blueberry seedlings are partially attributable to the regulation of phytohormone and sugar metabolism. This study provided valuable information for the research on the interaction between DSE fungi and host plants as well as the application of DSE preparations in agriculture.

Information System Model and Key Technologies of High-Definition Maps in Autonomous Driving Scenarios.

Background: High-definition maps can provide necessary prior data for autonomous driving, as well as the corresponding beyond-line-of-sight perception, verification and positioning, dynamic planning, and decision control. It is a necessary element to achieve L4/L5 unmanned driving at the current stage. However, currently, high-definition maps still have problems such as a large amount of data, a lot of data redundancy, and weak data correlation, which make autonomous driving fall into difficulties such as high data query difficulty and low timeliness. In order to optimize the data quality of high-definition maps, enhance the degree of data correlation, and ensure that they better assist vehicles in safe driving and efficient passage in the autonomous driving scenario, it is necessary to clarify the information system thinking of high-definition maps, propose a complete and accurate model, determine the content and functions of each level of the model, and continuously improve the information system model. Objective: The study aimed to put forward a complete and accurate high-definition map information system model and elaborate in detail the content and functions of each component in the data logic structure of the system model. Methods: Through research methods such as the modeling method and literature research method, we studied the high-definition map information system model in the autonomous driving scenario and explored the key technologies therein. Results: We put forward a four-layer integrated high-definition map information system model, elaborated in detail the content and functions of each component (map, road, vehicle, and user) in the data logic structure of the model, and also elaborated on the mechanism of the combined information of each level of the model to provide services in perception, positioning, decision making, and control for autonomous driving vehicles. This article also discussed two key technologies that can support autonomous driving vehicles to complete path planning, navigation decision making, and vehicle control in different autonomous driving scenarios. Conclusions: The four-layer integrated high-definition map information model proposed by this research institute has certain application feasibility and can provide references for the standardized production of high-definition maps, the unification of information interaction relationships, and the standardization of map data associations.

Deep learning image reconstruction for low-kiloelectron volt virtual monoenergetic images in abdominal dual-energy CT: medium strength provides higher lesion conspicuity.

BACKGROUND: The best settings of deep learning image reconstruction (DLIR) algorithm for abdominal low-kiloelectron volt (keV) virtual monoenergetic imaging (VMI) have not been determined. PURPOSE: To determine the optimal settings of the DLIR algorithm for abdominal low-keV VMI. MATERIAL AND METHODS: The portal-venous phase computed tomography (CT) scans of 109 participants with 152 lesions were reconstructed into four image series: VMI at 50 keV using adaptive statistical iterative reconstruction (Asir-V) at 50% blending (AV-50); and VMI at 40 keV using AV-50 and DLIR at medium (DLIR-M) and high strength (DLIR-H). The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of nine anatomical sites were calculated. Noise power spectrum (NPS) using homogenous region of liver, and edge rise slope (ERS) at five edges were measured. Five radiologists rated image quality and diagnostic acceptability, and evaluated the lesion conspicuity. RESULTS: The SNR and CNR values, and noise and noise peak in NPS measurements, were significantly lower in DLIR images than AV-50 images in all anatomical sites (all P < 0.001). The ERS values were significantly higher in 40-keV images than 50-keV images at all edges (all P < 0.001). The differences of the peak and average spatial frequency among the four reconstruction algorithms were significant but relatively small. The 40-keV images were rated higher with DLIR-M than DLIR-H for diagnostic acceptance (P < 0.001) and lesion conspicuity (P = 0.010). CONCLUSION: DLIR provides lower noise, higher sharpness, and more natural texture to allow 40 keV to be a new standard for routine VMI reconstruction for the abdomen and DLIR-M gains higher diagnostic acceptance and lesion conspicuity rating than DLIR-H.

Deep learning image reconstruction generates thinner slice iodine maps with improved image quality to increase diagnostic acceptance and lesion conspicuity: a prospective study on abdominal dual-energy CT.

BACKGROUND: To assess the improvement of image quality and diagnostic acceptance of thinner slice iodine maps enabled by deep learning image reconstruction (DLIR) in abdominal dual-energy CT (DECT). METHODS: This study prospectively included 104 participants with 136 lesions. Four series of iodine maps were generated based on portal-venous scans of contrast-enhanced abdominal DECT: 5-mm and 1.25-mm using adaptive statistical iterative reconstruction-V (Asir-V) with 50% blending (AV-50), and 1.25-mm using DLIR with medium (DLIR-M), and high strength (DLIR-H). The iodine concentrations (IC) and their standard deviations of nine anatomical sites were measured, and the corresponding coefficient of variations (CV) were calculated. Noise-power-spectrum (NPS) and edge-rise-slope (ERS) were measured. Five radiologists rated image quality in terms of image noise, contrast, sharpness, texture, and small structure visibility, and evaluated overall diagnostic acceptability of images and lesion conspicuity. RESULTS: The four reconstructions maintained the IC values unchanged in nine anatomical sites (all p > 0.999). Compared to 1.25-mm AV-50, 1.25-mm DLIR-M and DLIR-H significantly reduced CV values (all p < 0.001) and presented lower noise and noise peak (both p < 0.001). Compared to 5-mm AV-50, 1.25-mm images had higher ERS (all p < 0.001). The difference of the peak and average spatial frequency among the four reconstructions was relatively small but statistically significant (both p < 0.001). The 1.25-mm DLIR-M images were rated higher than the 5-mm and 1.25-mm AV-50 images for diagnostic acceptability and lesion conspicuity (all P < 0.001). CONCLUSIONS: DLIR may facilitate the thinner slice thickness iodine maps in abdominal DECT for improvement of image quality, diagnostic acceptability, and lesion conspicuity.

Three exonic variants in the PHEX gene cause aberrant splicing in a minigene assay.

Background: X-linked hypophosphatemia (XLH, OMIM 307800) is a rare phosphorus metabolism disorder caused by PHEX gene variants. Many variants simply classified as missense or nonsense variants were only analyzed at the DNA level. However, growing evidence indicates that some of these variants may alter pre-mRNA splicing, causing diseases. Therefore, this study aimed to use bioinformatics tools and a minigene assay to ascertain the effects of PHEX variations on pre-mRNA splicing. Methods: We analyzed 174 variants in the PHEX gene described as missense or nonsense variants. Finally, we selected eight candidate variants using bioinformatics tools to evaluate their effects on pre-mRNA splicing using a minigene assay system. The complementary DNA (cDNA) sequence for the PHEX gene (RefSeq NM_000444.6) serves as the basis for DNA variant numbering. Results: Of the eight candidate variants, three were found to cause abnormal splicing. Variants c.617T>G p.(Leu206Trp) and c.621T>A p.(Tyr207*) in exon 5 altered the splicing of pre-mRNA, owing to the activation of a cryptic splice site in exon 5, which produced an aberrant transcript lacking a part of exon 5, whereas variant c.1700G>C p.(Arg567Pro) in exon 16 led to the activation of a cryptic splice site in intron 16, resulting in a partial inclusion of intron 16. Conclusion: Our study employed a minigene system, which has a great degree of flexibility to assess abnormal splicing patterns under the circumstances of patient mRNA samples that are not available, to explore the impact of the exonic variants on pre-mRNA splicing. Based on the aforementioned experimental findings, we demonstrated the importance of analyzing exonic variants at the mRNA level.

Investigation of N-Glycan Functions in Receptor for Advanced Glycation End Products V Domain through Chemical Glycoprotein Synthesis.

The receptor for advanced glycation end products (RAGE) plays a crucial role in inflammation-related pathways and various chronic diseases. Despite the recognized significance of N-glycosylation in the ligand-binding V domain (VD) of RAGE, a comprehensive understanding of the site-activity and structure-activity relationships is lacking due to the challenges in obtaining homogeneous glycoprotein samples through biological expression. Here, we combined chemical and chemoenzymatic approaches to synthesize RAGE-VD and its congeners with Asn3-glycosylation by incorporating precise N-glycan structures. Evaluation of these samples revealed that, in comparison to other RAGE-VD forms, α2,6-sialylated N-glycosylation at the Asn3 site results in more potent inhibition of HMGB1-induced nuclear factor-κB (NF-κB) expression in RAGE-overexpressing cells. Hydrogen/deuterium exchange-mass spectrum analysis revealed a sialylated RAGE-VD-induced interaction region within HMGB1. Conversely, Asn3 N-glycosylation in VD has negligible effects on RAGE-VD/S100B interactions. This study established an approach for accessing homogeneously glycosylated RAGE-VD and explored the modulatory effects of N-glycosylation on the interactions between RAGE-VD and its ligand proteins.

Identification of seven variants in the col4a1 gene that alter RNA splicing by minigene assay.

Type IV collagen is an integral component of basement membranes. Mutations in COL4A1, one of the key genes encoding Type IV collagen, can result in a variety of diseases. It is clear that a significant proportion of mutations that affect splicing can cause disease directly or contribute to the susceptibility or severity of disease. Here, we analyzed exonic mutations and intronic mutations described in the COL4A1 gene using bioinformatics programs and identified candidate mutations that may alter the normal splicing pattern through a minigene system. We identified seven variants that induce splicing alterations by disrupting normal splice sites, creating new ones, or altering splice regulatory elements. These mutations are predicted to impact protein function. Our results help in the correct molecular characterization of variants in COL4A1 and may help develop more personalized treatment options.

A novel heterozygous variant of the SALL1 gene with atypical Townes-Brocks syndrome phenotypes in Chinese family.

Townes-Brocks syndrome (TBS) is an autosomal dominant disorder characterised by the triad of anorectal, thumb, and ear malformations. It may also be accompanied by defects in kidney, heart, eyes, hearing, and feet. TBS has been demonstrated to result from heterozygous variants in the SALL1 gene, which encodes zinc finger protein believed to function as a transcriptional repressor. The clinical characteristics of an atypical TBS phenotype patient from a Chinese family are described, with predominant manifestations including external ear dysplasia, unilateral renal hypoplasia with mild renal dysfunction, and hearing impairment. A novel heterozygous variant c.3060T>A (p.Tyr1020*) in exon 2 of the SALL1 gene was identified in this proband. Pyrosequencing of the complementary DNA of the proband revealed that the variant transcript accounted for 48% of the total transcripts in peripheral leukocytes, indicating that this variant transcript has not undergone nonsense-mediated mRNA decay. This variant c.3060T > A is located at the terminal end of exon 2, proximal to the 3' end of the SALL1 gene, and exerts a relatively minor impact on protein function. We suggest that the atypical TBS phenotype observed in the proband may be attributed to the truncated protein retaining partial SALL1 function.

Valosin-containing protein acts as a target and mediator of S-nitrosylation in the heart through distinct mechanisms.

S-nitrosylation (SNO) is an emerging paradigm of redox signaling protecting cells against oxidative stress in the heart. Our previous studies demonstrated that valosin-containing protein (VCP), an ATPase-associated protein, is a vital mediator protecting the heart against cardiac stress and ischemic injury. However, the molecular regulations conferred by VCP in the heart are not fully understood. In this study, we explored the potential role of VCP in cardiac protein SNO using multiple cardiac-specific genetically modified mouse models and various analytical techniques including biotin switch assay, liquid chromatography, mass spectrometry, and western blotting. Our results showed that cardiac-specific overexpression of VCP led to an overall increase in the levels of SNO-modified cardiac proteins in the transgenic (TG) vs. wild-type (WT) mice. Mass spectrometry analysis identified mitochondrial proteins involved in respiration, metabolism, and detoxification as primary targets of SNO modification in VCP-overexpressing mouse hearts. Particularly, we found that VCP itself underwent SNO modification at a specific cysteine residue in its N-domain. Additionally, our study demonstrated that glyceraldehyde 3-phosphate dehydrogenase (GAPDH), a key enzyme in glycolysis, also experienced increased SNO in response to VCP overexpression. While deletion of inducible nitric oxide synthase (iNOS) in VCP TG mice did not affect VCP SNO, it did abolish SNO modification in mitochondrial complex proteins, suggesting a dual mechanism of regulation involving both iNOS-dependent and independent pathways. Overall, our findings shed light on post-translational modification of VCP in the heart, unveiling a previously unrecognized role for VCP in regulating cardiac protein SNO and offering new insights into its function in cardiac protection.

Impact of Postoperative Radiotherapy on the Prognosis of Early-Stage (pT1-2N0M0) Oral Tongue Squamous Cell Carcinoma.

PURPOSE: To identify subgroups of patients with early-stage (pT1-2N0M0) oral tongue squamous cell carcinoma (OTSCC) who may benefit from postoperative radiotherapy (PORT). PATIENTS AND METHODS: This retrospective cohort study included 528 patients diagnosed between October 2009 and December 2021. Clinicopathological characteristics and treatments with or without PORT were analyzed for their impact on outcomes. RESULTS: Among 528 patients who underwent radical surgery (median age, 62 years [IQR, 52-69]), 145 (27.5%) also underwent PORT. Multivariate analyses revealed that PORT was associated with improved survival outcomes, whereas moderate-to-poor differentiation, perineural infiltration (PNI), lymphovascular invasion (LVI), and increasing depth of invasion (DOI) were associated with poorer survival outcomes. For patients with moderate-to-poor differentiation, the surgery + PORT group showed improved outcomes compared with the surgery-alone group. After propensity score matching, the results were as follows: overall survival (OS), 97% versus 69%, P = .003; disease-free survival (DFS), 88% versus 50%, P = .001. After excluding cases with PNI/LVI, the differences persisted: OS, 97% versus 82%, P = .040; DFS, 87% versus 64%, P = .012. Similar survival benefits were observed in 104 patients with PNI and/or LVI (OS, 81% v 58%; P = .022; DFS, 76% v 47%; P = .002). In subgroups with DOI >5 mm or close margins, PORT contributed to improved DFS (80% v 64%; P = .006; 92% v 66%; P = .049) but did not significantly affect OS. CONCLUSION: Patients with moderately-to-poorly differentiated pT1-2N0M0 OTSCC benefited from PORT. Our study provided evidence that patients with PNI and/or LVI who underwent PORT had improved survival. PORT also offered DFS benefit among patients with DOI >5 mm.

Tracking endogenous proteins based on RNA editing-mediated genetic code expansion.

Protein labeling approaches are important to study proteins in living cells, and genome editing tools make it possible to tag endogenous proteins to address the concerns associated with overexpression. Here we established RNA editing-mediated noncanonical amino acids (ncAAs) protein tagging (RENAPT) to site-specifically label endogenous proteins with ncAAs in living cells. RENAPT labels protein in a temporary and nonheritable manner and is not restricted by protospacer adjacent motif sequence. Using a fluorescent ncAA or ncAA with a bio-orthogonal reaction handle for subsequent dye labeling, we demonstrated that a variety of endogenous proteins can be imaged at their specific subcellular locations. In addition, two proteins can be tagged individually and simultaneously using two different ncAAs. Furthermore, endogenous ion channels and neuron-specific proteins can be real-time labeled in primary neurons. Thus, RENAPT presents a promising platform with broad applicability for tagging endogenous proteins in living cells to study their localization and functions.

Impacts of Adaptive Statistical Iterative Reconstruction-V and Deep Learning Image Reconstruction Algorithms on Robustness of CT Radiomics Features: Opportunity for Minimizing Radiomics Variability Among Scans of Different Dose Levels.

This study aims to investigate the influence of adaptive statistical iterative reconstruction-V (ASIR-V) and deep learning image reconstruction (DLIR) on CT radiomics feature robustness. A standardized phantom was scanned under single-energy CT (SECT) and dual-energy CT (DECT) modes at standard and low (20 and 10 mGy) dose levels. Images of SECT 120 kVp and corresponding DECT 120 kVp-like virtual monochromatic images were generated with filtered back-projection (FBP), ASIR-V at 40% (AV-40) and 100% (AV-100) blending levels, and DLIR algorithm at low (DLIR-L), medium (DLIR-M), and high (DLIR-H) strength levels. Ninety-four features were extracted via Pyradiomics. Reproducibility of features was calculated between standard and low dose levels, between reconstruction algorithms in reference to FBP images, and within scan mode, using intraclass correlation coefficient (ICC) and concordance correlation coefficient (CCC). The average percentage of features with ICC > 0.90 and CCC > 0.90 between the two dose levels was 21.28% and 20.75% in AV-40 images, and 39.90% and 35.11% in AV-100 images, respectively, and increased from 15.43 to 45.22% and from 15.43 to 44.15% with an increasing strength level of DLIR. The average percentage of features with ICC > 0.90 and CCC > 0.90 in reference to FBP images was 26.07% and 25.80% in AV-40 images, and 18.88% and 18.62% in AV-100 images, respectively, and decreased from 27.93 to 17.82% and from 27.66 to 17.29% with an increasing strength level of DLIR. DLIR and ASIR-V algorithms showed low reproducibility in reference to FBP images, while the high-strength DLIR algorithm provides an opportunity for minimizing radiomics variability due to dose reduction.

Three exonic variants in the COL4A5 gene alter RNA splicing in a minigene assay.

BACKGROUND: X-linked Alport syndrome (XLAS) is an inherited renal disease caused by rare variants of COL4A5 on chromosome Xq22. Many studies have indicated that single nucleotide variants (SNVs) in exons can disrupt normal splicing process of the pre-mRNA by altering various splicing regulatory signals. The male patients with XLAS have a strong genotype-phenotype correlation. Confirming the effect of variants on splicing can help to predict kidney prognosis. This study aimed to investigate whether single nucleotide substitutions, located within three bases at the 5' end of the exons or internal position of the exons in COL4A5 gene, cause aberrant splicing process. METHODS: We analyzed 401 SNVs previously presumed missense and nonsense variants in COL4A5 gene by bioinformatics programs and identified candidate variants that may affect the splicing of pre-mRNA via minigene assays. RESULTS: Our study indicated three of eight candidate variants induced complete or partial exon skipping. Variants c.2678G>C and c.2918G>A probably disturb classic splice sites leading to corresponding exon skipping. Variant c.3700C>T may disrupt splicing enhancer motifs accompanying with generation of splicing silencer sequences resulting in the skipping of exon 41. CONCLUSION: Our study revealed that two missense variants positioned the first nucleotides of the 5' end of COL4A5 exons and one internal exonic nonsense variant caused aberrant splicing. Importantly, this study emphasized the necessity of assessing the effects of SNVs at the mRNA level.

Biosynthesis of Epipyrone A Reveals a Highly Specific Membrane-Bound Fungal C-Glycosyltransferase for Pyrone Galactosylation.

Epipyrone A is a unique C-galactosylated 4-hydroxy-2-pyrone derivative with an antifungal potential from the fungus Epicoccum nigrum. We elucidated its biosynthesis via heterologous expression and characterized an unprecedented membrane-bound pyrone C-glycosyltransferase biochemically. Molecular docking and mutagenesis experiments suggested a possible mechanism for the heterocyclic C-glycosylation and the importance of a transmembrane helix for its catalysis. These results expand the repertoire of C-glycosyltransferases and provide new insights into the formation of C-glycosides in fungi.

Potential Biocontrol Microorganisms Causing Attenuated Pathogenicity in Plasmopara viticola.

A phenomenon of pathogenicity attenuation of Plasmopara viticola was consistently observed during its subculture on grape. To clarify the causes of attenuated pathogenicity of P. viticola, culturable microbes were isolated from the P. viticola mass (mycelia, sporangiophores, and sporangia) in each generation and tested for their biocontrol efficacies on grape downy mildew (GDM). The results showed that the incidence of GDM decreased with the increase in the number of subculture times on both vineyard-collected leaves and grape leaves from in vitro-grown seedlings. The number of culturable microbial taxa on the surface of P. viticola decreased, whereas the population densities of four specific strains (i.e., K2, K7, P1, and P5) increased significantly with the increase in subculture times. Compared with the control, the biocontrol efficacies of the bacterial strain K2 reached 87.5%, and those of both fungal strains P1 and P5 reached 100.0%. Based on morphological characteristics and molecular sequences, strains K2, P1, and P5 were identified as Curtobacterium herbarum, Thecaphora amaranthi, and Acremonium sclerotigenum, respectively, and these three strains survived very well and multiplied on the surface of P. viticola. As the number of times P. viticola was subcultured increased, all three of these strains became the predominant strains, leading to greater P. viticola inhibition, attenuated P. viticola pathogenicity, and effective GDM biological control. To the best of our knowledge, this is the first report of C. herbarum and T. amaranthi having biological control activity against GDM.

Analyzing the Correlation Between Serum Biomarkers and Child-Pugh Score in Liver Cirrhosis.

Objective: This study aimed to analyze the diagnostic efficacy of serum biomarkers in liver cirrhosis patients categorized by Child-Pugh scores. Methods: An observational cross-sectional study design was employed. A total of 110 liver cirrhosis patients, classified according to Child-Pugh scores and 60 healthy individuals were included in this study. Serum levels of adenosine deaminase (ADA), adiponectin (APN), matrix metalloproteinase-2 (MMP-2), alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were measured. Results: The levels of ADA, APN, MMP-2, ALP, ALT, and AST were significantly higher in the study group compared to the control group (P < .05). Furthermore, these levels increased with the severity of liver cirrhosis, with higher levels observed in patients with Child-Pugh class C. The positive diagnostic rates for joint detection in Child-Pugh class A, B, and C were 93.75% (30/32), 100% (34/34), and 100% (44/44), respectively. Conclusions: Combined detection of serum biomarkers improves the diagnostic efficacy of liver cirrhosis. The diagnostic rates were higher when considering Child-Pugh scores, with the highest rates observed in class C.

Functional analysis of the CTNS gene exonic variants predicted to affect splicing.

Cystinosis is a severe, monogenic systemic disease caused by variants in CTNS gene. Currently, there is growing evidence that exonic variants in many diseases can affect pre-mRNA splicing. The impact of CTNS gene exonic variants on splicing regulation may be underestimated due to the lack of routine studies at the RNA level. Here, we analyzed 59 exonic variants in the CTNS gene using bioinformatics tools and identified candidate variants that may induce splicing alterations by minigene assays. We identified six exonic variants that induce splicing alterations by disrupting the ratio of exonic splicing enhancers/exonic splicing silencers (ESEs/ESSs) or by interfering with the recognition of classical splice sites, or both. Our results help in the correct molecular characterization of variants in cystinosis and inform emerging therapies. Furthermore, our work suggests that the combination of in silico and in vitro assays facilitates to assess the effects of DNA variants driving rare genetic diseases on splicing regulation and will enhance the clinical utility of variant functional annotation.