Insights into pollution characteristics and human health risks of plasticizer phthalate esters in shellfish species.

The ubiquitous application of phthalate esters (PAEs) as plasticizers contributes to high levels of marine pollution, yet the contamination patterns of PAEs in various shellfish species remain unknown. The objective of this research is to provide the first information on the pollution characteristics of 16 PAEs in different shellfish species from the Pearl River Delta (PRD), South China, and associated health risks. Among the 16 analyzed PAEs, 13 were identified in the shellfish, with total PAE concentrations ranging from 23.07 to 3794.08 ng/g dw (mean = 514.35 ng/g dw). The PAE pollution levels in the five shellfish species were as follows: Ostreidae (mean = 1064.12 ng/g dw) > Mytilus edulis (mean = 509.88 ng/g dw) > Babylonia areolate (mean = 458.14 ng/g dw) > Mactra chinensis (mean = 378.90 ng/g dw) > Haliotis diversicolor (mean = 335.28 ng/g dw). Dimethyl phthalate (DMP, mean = 69.85 ng/g dw), diisobutyl phthalate (DIBP, mean = 41.39 ng/g dw), dibutyl phthalate (DBP, mean = 130.91 ng/g dw), and di(2-ethylhexyl) phthalate (DEHP, mean = 226.23 ng/g dw) were the most abundant congeners. Notably, DEHP constituted the most predominant fraction (43.98 %) of the 13 PAEs detected in all shellfish from the PRD. Principal component analysis indicated that industrial and domestic emissions served as main sources for the PAE pollution in shellfish from the PRD. It was estimated that the daily intake of PAEs via shellfish consumption among adults and children ranged from 0.004 to 1.27 µg/kgbw/day, without obvious non-cancer risks (< 0.034), but the cancer risks raised some alarm (2.0 × 10-9-1.4 × 10-5). These findings highlight the necessity of focusing on marine environmental pollutants and emphasize the importance of ongoing monitoring of PAE contamination in seafood.

Electrical stress and acid orange 7 synergistically clear the blockage of electron flow in the methanogenesis of low-strength wastewater.

Energy recovery from low-strength wastewater through anaerobic methanogenesis is constrained by limited substrate availability. The development of efficient methanogenic communities is critical but challenging. Here we develop a strategy to acclimate methanogenic communities using conductive carrier (CC), electrical stress (ES), and Acid Orange 7 (AO7) in a modified biofilter. The synergistic integration of CC, ES, and AO7 precipitated a remarkable 72-fold surge in methane production rate compared to the baseline. This increase was attributed to an altered methanogenic community function, independent of the continuous presence of AO7 and ES. AO7 acted as an external electron acceptor, accelerating acetogenesis from fermentation intermediates, restructuring the bacterial community, and enriching electroactive bacteria (EAB). Meanwhile, CC and ES orchestrated the assembly of the archaeal community and promoted electrotrophic methanogens, enhancing acetotrophic methanogenesis electron flow via a mechanism distinct from direct electrochemical interactions. The collective application of CC, ES, and AO7 effectively mitigated electron flow impediments in low-strength wastewater methanogenesis, achieving an additional 34% electron recovery from the substrate. This study proposes a new method of amending anaerobic digestion systems with conductive materials to advance wastewater treatment, sustainability, and energy self-sufficiency.

VirGrapher: a graph-based viral identifier for long sequences from metagenomes.

Viruses are the most abundant biological entities on earth and are important components of microbial communities. A metagenome contains all microorganisms from an environmental sample. Correctly identifying viruses from these mixed sequences is critical in viral analyses. It is common to identify long viral sequences, which has already been passed thought pipelines of assembly and binning. Existing deep learning-based methods divide these long sequences into short subsequences and identify them separately. This makes the relationships between them be omitted, leading to poor performance on identifying long viral sequences. In this paper, VirGrapher is proposed to improve the identification performance of long viral sequences by constructing relationships among short subsequences from long ones. VirGrapher see a long sequence as a graph and uses a Graph Convolutional Network (GCN) model to learn multilayer connections between nodes from sequences after a GCN-based node embedding model. VirGrapher achieves a better AUC value and accuracy on validation set, which is better than three benchmark methods.

Acceleration of the ocean warming from 1961 to 2022 unveiled by large-ensemble reanalyses.

Long-term changes in ocean heat content (OHC) represent a fundamental global warming indicator and are mostly caused by anthropogenic climate-altering gas emissions. OHC increases heavily threaten the marine environment, therefore, reconstructing OHC before the well-instrumented period (i.e., before the Argo floats deployment in the mid-2000s) is crucial to understanding the multi-decadal climate change in the ocean. Here, we shed light on ocean warming and its uncertainty for the 1961-2022 period through a large ensemble reanalysis system that spans the major sources of uncertainties. Results indicate a 62-year warming of 0.43 ± 0.08 W m-2, and a statistically significant acceleration rate equal to 0.15 ± 0.04 W m-2 dec-1, locally peaking at high latitudes. The 11.6% of the global ocean area reaches the maximum yearly OHC in 2022, almost doubling any previous year. At the regional scale, major OHC uncertainty is found in the Tropics; at the global scale, the uncertainty represents about 40% and 15% of the OHC variability, respectively before and after the mid-2000s. The uncertainty of regional trends is mostly affected by observation calibration (especially at high latitudes), and sea surface temperature data uncertainty (especially at low latitudes).

Electrical Conductivity and pH Are Two of the Main Factors Influencing the Composition of Arbuscular Mycorrhizal Fungal Communities in the Vegetation Succession Series of Songnen Saline-Alkali Grassland.

Arbuscular mycorrhizal fungi (AMF) are widely distributed microorganisms in the soil, playing an important role in vegetation succession, plant community diversity, and improving soil physicochemical properties. In this study, morphological identification and high-throughput sequencing technology were used to comprehensively analyze the AMF community composition and diversity at different succession stages of Songnen saline-alkali grassland. To determine the root colonization status of plants collected in the field, a colonization system was established using late-succession plants as host plants to verify the existence of mycorrhizal symbiosis and the matching phenomenon of AMF in Songnen saline-alkali grassland. The results indicated that both morphological methods and high-throughput sequencing technology showed that glomus was the dominant genus of AMF in Songnen saline grassland. Redundancy analysis (RDA) and linear regression analysis showed that electrical conductivity (EC) and pH were the main environmental factors affecting AMF species diversity and community structure in the succession sequence of Songnen saline grassland. In addition, the results of root colonization identification and the colonization system test in the field showed that AMF successfully colonized vegetation at different succession stages and had mycorrhizal symbiosis. The results of this study could help to understand the AMF community of Songnen saline-alkali grassland as well as provide a reference and basis for optimizing the AMF community structure of Songnen saline-alkali grassland through human intervention in the future and using mycorrhizal technology to restore and rebuild the degraded ecosystem of Songnen saline-alkali grassland.

Occurrence and distribution of polycyclic aromatic hydrocarbons (PAHs) in marine organisms from Shenzhen coastal waters and human health risk assessment.

This study investigated the levels and composition of 16 priority polycyclic aromatic hydrocarbons (PAHs) in marine organisms from South China Sea and assessed their potential health risks. The results revealed that the pollution levels of total PAHs ranged from 3.56 to 392.21 ng/g dw. Notably, 4-ring PAHs constituted the predominant fraction (58.02 %) of the total PAHs, with pyrene being the most abundant congener across all species. Intriguingly, our findings suggested that consuming these organisms might pose a low non-cancer hazard. Nonetheless, benzo[a]pyrene was detected in most species, with levels ranging from non-detectable to 11.24 ng/g dw. The individual lifetime cancer risk levels associated with seafood consumption in studied regions ranged from 1.10 × 10-5 to 1.52 × 10-5, highlighting a potential cancer risk that warrants special attention. These findings emphasize the need to prioritize carcinogenic compounds over total PAHs and underscore the importance of continuous monitoring of PAH pollution in seafood.

Establishment and Validation of Risk Prediction Model for Postpartum Hemorrhage for Pregnant Women ≥35 Years of Age in Natural Delivery.

Context: After the age of 35, women's fertility and physical function gradually decline, and this can significantly increase the risks of postpartum hemorrhage (PPH) after delivery. Sufficient exploration of prenatal indicators of PPH for older pregnant women are still lacking. Objective: The study intended to examine the factors influencing postpartum hemorrhage (PPH) in natural delivery for pregnant women ≥35 years of age and to establish a reliable risk-prediction model. Design: The research team performed a retrospective study. Setting: The study took place at Suzhou Ninth People's Hospital in Suzhou, Jiangsu Province, China. Participants: Participants were 351 pregnant women who had undergone a prenatal examination and vaginal delivery at the hospital between January 2019 and October 2022. Groups: The research team divided participants into two groups: (1) a PPH group, with 52 participants who experienced PPH, and (2) a non-PPH group, with 299 participants who had no PPH. Outcome Measures: The research team: (1) conducted single-factor analysis of the two groups' demographic and clinical characteristics; (2) performed multivariate logistic regression analysis to find the factors influencing PPH; (3) built a risk-prediction model based on the results; and (4) analyzed the model's identification ability, proofreading ability, and clinical applicability using a goodness-of-fit test, a receiver operating characteristic (ROC) curve, a calibration curve, and a decision curve. The team used the SPSS 22.0 and R software for statistical analysis. Results: The incidence of PPH was 14.81%, for the 52 out of 351 participants. The PPH group's age (P < .001), rate of pregnancy-induced hypertension (P = .008), length of the third stage of labor (P = .001), and newborn's birth weight (P < .001) were significantly greater and its FIB before delivery was significantly lower than those of the non-PPH group. The high expression of fibrinogen (FIB) before delivery indicates it may be a protective factor against PPH. The multivariate analysis indicated that a greater age (P = .013), pregnancy-induced hypertension (P = .002), a low FIB level (P < .001), a long third stage of labor (P = .012), and a low birth weight for the newborn (P = .006) were all significantly related to PPH. The research team validated the risk-prediction model, which indicated that the model had good recognition ability (AUC = 0.873). The optimal critical value was 34%, and the sensitivity and specificity were 0.869 and 0.826, respectively. In the comparison of the PPH value that the model predicted and the participants' actual PPH incidence (U = -0.006, Brier = 0.089), the deviation of the model wasn't statistically significant (χ2 = 5.964, P = .651). The analysis of the decision curve found that the net benefits for pregnant women ≥35 years of age were higher than those of the other two extreme curves, showing that the model was clinically effective. Conclusions: The PPH risk-prediction model for vaginal delivery for pregnant women ≥35 years of age showed that a greater age, pregnancy-induced hypertension, a lower prelabor FIB, a longer third stage of labor, and a higher birth weight for the newborn were significantly related to the incidence of PPH and that its use could be clinically helpful.

Three-dimensional magnetic resonance elastography combining proton-density fat fraction precisely identifies metabolic dysfunction-associated steatohepatitis with significant fibrosis.

PURPOSE: Patients with metabolic dysfunction-associated steatohepatitis (MASH) and significant fibrosis (fibrosis stage≥2), known as Fibro-MASH, are at increased risk of liver-related outcomes and lower rates of spontaneous disease regression. The aim was to investigate three-dimensional MR elastography (3D-MRE) combining proton-density fat fraction (PDFF) as a means of identifying Fibro-MASH. METHODS: Forty-eight New Zealand rabbits were fed a high-fat/cholesterol or standard diet to obtain different disease activity and fibrosis stages. Shear stiffness (SS) and Damping Ratio (DR) were derived from 3D-MRE, whereas PDFF was from a volumetric 3D imaging sequence. Steatosis grade, metabolic dysfunction-associated steatotic liver disease activity score (MAS), and fibrosis stage were diagnosed histologically. Serum markers of fibrosis and inflammation were also measured. Correlation and comparison analysis, Receiver operating characteristic curves (ROC), Delong test, logistic regression analysis, and Net reclassification improvement (NRI) were performed. RESULTS: PDFF correlated with steatosis grade (rho = 0.853). SS increased with developed liver fibrosis (rho = 0.837). DR correlated with MAS grade (rho = 0.678). The areas under the ROC (AUROCs) of SS for fibrosis grading were 0.961 and 0.953 for ≥F2, and ≥ F3, respectively. All the biochemical parameters were considered but excluded from the logistic regression analysis to identify Fibro-MASH. FF, SS, and DR were finally included in the further analysis. The three-parameter model combining PDFF, SS, and DR showed significant improvement in NRI over the model combining SS and PDFF (AUROC 0.973 vs. 0.906, P = 0.081; NRI 0.28, P < 0.05). CONCLUSION: 3D-MRE combining PDFF may characterize the state of fat content, disease activity and fibrosis, thus precisely identify Fibro-MASH.

NF-κB signaling pathway mechanism in cow intertoe skin inflammation caused by Fusobacterium necrophorum.

Background: Fusobacterium necrophorum is the main pathogen inducing bovine foot rot. The infected site is often accompanied by a strong inflammatory response, but the specific inflammatory regulatory mechanism remains unclear. Aim: A cow skin explants model was established to elucidate the mechanism of F. necrophorum bacillus causing foot rot in cows, and to provide reference for future clinical practice. Methods: Cow intertoe skin explants were cultured in vitro, and F. necrophorum bacteria solution and nuclear factor-κB (NF-κB) inhibitor BAY 1-7082 were added to establish an in vitro infection model. Hematoxylin and eosin staining, terminal - deoxynucleotidyl transferase mediated nick end labeling, and immunohistochemistry were used to detect the pathological changes of the skin explants infected with F. necrophorum, the degree of tissue cell apoptosis, and the expression of the apoptosis-related protein Caspase-3, respectively. RT-qPCR, Western blot, and ELISA were used to detect the activation of the NF-κB pathway and inflammatory cytokines by F. necrophorum. Results: The intertoe skin structure of cows infected with F. necrophorum changed with different degrees of inflammation, and the degree of tissue cell apoptosis was significantly increased (P < 0.01). In addition, infection with F. necrophorum significantly increased the phosphorylation level of IκBα protein and up-regulated the expression level of NF-κB p65. The high expression and transcriptional activity of NF-κB p65 significantly increased the expression and concentration of the inflammatory cytokines TNF-α, IL-1ß, and IL-8, thus inducing the occurrence of an inflammatory response. However, inhibition of NF-κB p65 activity significantly decreased the expression of inflammatory factors in the intertoe skin of cows infected with F. necrophorum. Conclusion: F. necrophorum activates NF-κB signaling pathway by increasing the expression of TNF-α, IL-1ß, IL-8 and other inflammatory factors, leading to foot rot in dairy cows.

Hypoxia-inducible factor orchestrates adenosine metabolism to promote liver cancer development.

Hypoxia-induced adenosine creates an immunosuppressive tumor microenvironment (TME) and dampens the efficacy of immune checkpoint inhibitors (ICIs). We found that hypoxia-inducible factor 1 (HIF-1) orchestrates adenosine efflux through two steps in hepatocellular carcinoma (HCC). First, HIF-1 activates transcriptional repressor MXI1, which inhibits adenosine kinase (ADK), resulting in the failure of adenosine phosphorylation to adenosine monophosphate. This leads to adenosine accumulation in hypoxic cancer cells. Second, HIF-1 transcriptionally activates equilibrative nucleoside transporter 4, pumping adenosine into the interstitial space of HCC, elevating extracellular adenosine levels. Multiple in vitro assays demonstrated the immunosuppressive role of adenosine on T cells and myeloid cells. Knockout of ADK in vivo skewed intratumoral immune cells to protumorigenic and promoted tumor progression. Therapeutically, combination treatment of adenosine receptor antagonists and anti-PD-1 prolonged survival of HCC-bearing mice. We illustrated the dual role of hypoxia in establishing an adenosine-mediated immunosuppressive TME and offered a potential therapeutic approach that synergizes with ICIs in HCC.

Characterization and distribution of polybrominated diphenyl ethers in shellfish in Shenzhen coastal waters and assessment of human health risks.

This study aims to investigate the profiles of polybrominated diphenyl ethers (PBDEs) in shellfish obtained from Shenzhen coastal waters and assess the potential health risks. We analyzed 74 shellfish samples from eight different species for PBDEs (BDE-28, -47, -99, -100, -153, -154, -183, -209). The concentrations of total PBDEs in different shellfish species ranged from 2.02 to 360.17 pg g-1 wet weight, with the highest levels found in Pectinidae, Babylonia areolate, Ostreidae, Perna viridis, Haliotis diversicolor, Corbiculidae, Pinctada margaritifera, and Veneridae in descending order. Among the PBDE congeners analyzed, BDE-47 was the most abundant, followed by BDE-154 and BDE-153. Furthermore, the estimated daily intake of PBDEs through shellfish consumption for Shenzhen residents were between 0.11 and 0.19 ng kg-1(bw) day-1. To our knowledge, this is the first study to systematically investigate the profiles of PBDEs in eight different shellfish species from Shenzhen's coastal waters and evaluate the potential human health risks associated with shellfish consumption.

Ferroptosis Suppressor Protein 1 Inhibition Promotes Tumor Ferroptosis and Anti-tumor Immune Responses in Liver Cancer.

BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) is a highly aggressive malignancy with dreadful clinical outcome. Tyrosine kinase inhibitors and immune checkpoint inhibitors are the only United States Food and Drug Administration-approved therapeutic options for patients with advanced HCC with limited therapeutic success. Ferroptosis is a form of immunogenic and regulated cell death caused by chain reaction of iron-dependent lipid peroxidation. Coenzyme Q10 (CoQ10)/ferroptosis suppressor protein 1 (FSP1) axis was recently identified as a novel protective mechanism against ferroptosis. We would like to explore whether FSP1 could be a potential therapeutic target for HCC. METHODS: FSP1 expression in human HCC and paired non-tumorous tissue samples were determined by reverse transcription-quantitative polymerase chain reaction, followed by clinicopathologic correlation and survival studies. Regulatory mechanism for FSP1 was determined using chromatin immunoprecipitation. The hydrodynamic tail vein injection model was used for HCC induction to evaluate the efficacy of FSP1 inhibitor (iFSP1) in vivo. Single-cell RNA sequencing revealed the immunomodulatory effects of iFSP1 treatment. RESULTS: We showed that HCC cells greatly rely on the CoQ10/FSP1 system to overcome ferroptosis. We found that FSP1 was significantly overexpressed in human HCC and is regulated by kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2 pathway. FSP1 inhibitor iFSP1 effectively reduced HCC burden and profoundly increased immune infiltrates including dendritic cells, macrophages, and T cells. We also demonstrated that iFSP1 worked synergistically with immunotherapies to suppress HCC progression. CONCLUSIONS: We identified FSP1 as a novel, vulnerable therapeutic target in HCC. The inhibition of FSP1 potently induced ferroptosis, which promoted innate and adaptive anti-tumor immune responses and effectively suppressed HCC tumor growth. FSP1 inhibition therefore represents a new therapeutic strategy for HCC.

Genome-Wide CRISPR/Cas9 Library Screening Revealed Dietary Restriction of Glutamine in Combination with Inhibition of Pyruvate Metabolism as Effective Liver Cancer Treatment.

Hepatocellular carcinoma (HCC) is the second most lethal cancer worldwide. Glutamine is an essential, extracellular nutrient which supports HCC growth. Dietary glutamine deficiency may be a potential therapeutic approach for HCC. HCC cells overcome metabolic challenges by rewiring their metabolic pathways for rapid adaptations. The efficiency of dietary glutamine deficiency as HCC treatment is examined and the adaptation machinery under glutamine depletion in HCC cells is unraveled. Using genome-wide CRISPR/Cas9 knockout library screening, this study identifies that pyruvate dehydrogenase α (PDHA), pyruvate dehydrogenase ß (PDHB), and pyruvate carboxylase (PC) in pyruvate metabolism are crucial to the adaptation of glutamine depletion in HCC cells. Knockout of either PDHA, PDHB or PC induced metabolic reprogramming of the tricarboxylic acid (TCA) cycle, disrupts mitochondrial function, leading to the suppression of HCC cell proliferation under glutamine depletion. Surprisingly, dietary glutamine restriction improves therapeutic responses of HCC to PDH or PC inhibitor in mouse HCC models. Stable isotope carbon tracing confirms that PDH or PC inhibitors further disrupt the metabolic rewiring of the TCA cycle induced by dietary glutamine depletion in HCC. In summary, the results demonstrate that pyruvate metabolism acts as novel targetable metabolic vulnerabilities for HCC treatment in combination with a glutamine-deficient diet.

A novel missense mutation in obscurin gene in a Chinese consanguineous family with left ventricular noncompaction.

BACKGROUND: Left ventricular noncompaction (LVNC) is an increasingly recognised cardiomyopathy of which a significant percentage are genetic in origin. The purpose of the present study was to identify potential pathogenic mutation leading to disease in a Chinese LVNC family. METHODS: A 3-generation family affected by LVNC was recruited. Clinical assessments were performed on available family members, with clinical examination, ECG, echocardiography and cardiac MRI. The proband (I-2), the proband's daughter (II-1, affected) and mother (III-1, unaffected) were selected for WGS. Sanger sequencing were performed in all of the 4 surviving family members. RESULTS: Combined whole genome sequencing with linkage analysis identified a novel missense mutation in the giant protein obscurin (OBSCN NM_001098623, c.C19063T), as the only plausible disease-causing variant that segregates with disease among the four surviving individuals, with interrogation of the entire genome excluding other potential causes. This c.C19063T missense mutation resulted in p.R6355W in the encoded OBSCN protein. It affected a highly conserved residue in the C terminus of the obscurin-B-like isoform between the PH and STKc domains, which was predicted to affect the function of the protein by different bioinformatics tools. CONCLUSIONS: Here we present clinical and genetic evidence implicating the novel R6355W missense mutation in obscurin as the cause of familial LVNC. This expands the spectrum of obscurin's roles in cardiomyopathies. It furthermore highlights that rare obscurin missense variants, currently often ignored or left uninterpreted, should be considered to be relevant for cardiomyopathies and can be identified by the approach presented here. This study also provided new insights into the molecular basis of OBSCN mutation positive LVNC.

Corrigendum: Pathogenicity and Long-Term Outcomes of Liddle Syndrome Caused by a Nonsense Mutation of SCNN1G in a Chinese Family.

[This corrects the article DOI: 10.3389/fped.2022.887214.].

Chronic effects of benzalkonium chlorides on short chain fatty acids and methane production in semi-continuous anaerobic digestion of waste activated sludge.

As an emerging pollutant, benzalkonium chlorides (BACs) potentially enriched in waste activated sludge (WAS). However, the microbial response mechanism under chronic effects of BACs on acidogenesis and methanogenesis in anaerobic digestion (AD) has not been clearly disclosed. This study investigated the AD (by-)products and microbial evolution under low to high BACs concentrations from bioreactor startup to steady running. It was found that BACs can lead to an increase of WAS hydrolysis and fermentation, but a disturbance to acidogenic bacteria also occurred at low BACs concentration. A noticeable inhibition to methanogenesis occurred when BAC concentration was up to 15 mg/g TSS. Metagenomic analysis revealed the key genes involved in acetic acid (HAc) biosynthesis (i.e. phosphate acetyltransferase, PTA), ß-oxidation pathway (acetyl-CoA C-acetyltransferase) and propionic acid (HPr) conversion was slightly promoted compared with control. Furthermore, BACs inhibited the acetotrophic methanogenesis (i.e. acetyl-CoA synthetase), especially BAC concentration was up to 15 mg/g TSS, thereby enhanced short chain fatty acids (SCFAs) accumulation. Overall, chronic stimulation of functional microorganisms with increasing concentrations of BACs impact WAS fermentation.

Tomoelastography based on multifrequency MR elastography predicts liver function reserve in patients with hepatocellular carcinoma: a prospective study.

BACKGROUND: Estimating liver function reserve is essential for preoperative surgical planning and predicting post-hepatectomy complications in patients with hepatocellular carcinoma (HCC). We investigated hepatic viscoelasticity quantified by tomoelastography, a multifrequency magnetic resonance elastography technique, to predict liver function reserve. METHODS: One hundred fifty-six patients with suspected HCC (mean age, 60 ± 1 years; 131 men) underwent preoperative tomoelastography examination between July 2020 and August 2021. Sixty-nine were included in the final analysis, and their 15-min indocyanine green retention rates (ICG-R15s) were obtained to determine liver function reserve. Tomoelastography quantified the shear wave speed (c, m/s), which represents stiffness, and loss angle (φ, rad), which represents fluidity. Both were correlated with the ICG-R15. A prediction model based on logistic regression for major hepatectomy tolerance (ICG-R15 ≥ 14%) was established. RESULTS: Patients were assigned to either the ICG-R15 < 14% (n = 50) or ICG-R15 ≥ 14% (n = 19) group. Liver c (r = 0.617) and φ (r = 0.517) were positively correlated with the ICG-R15 (both p < 0.001). At fibrosis stages F1-2, φ was positively correlated with the ICG-R15 (r = 0.528; p = 0.017), but c was not (p = 0.104). At stages F3-4, c (r = 0.642; p < 0.001) and φ (r = 0.377; p = 0.008) were both positively correlated with the ICG-R15. The optimal cutoffs of c and φ for predicting ICG-R15 ≥ 14% were 2.04 m/s and 0.79 rad, respectively. The area under the receiver operating characteristic curve was higher for c (0.892) than for φ (0.779; p = 0.045). CONCLUSIONS: Liver stiffness and fluidity, quantified by tomoelastography, were correlated with liver function and may be used clinically to noninvasively assess liver function reserve and stratify treatments.

Pathogenicity and Long-Term Outcomes of Liddle Syndrome Caused by a Nonsense Mutation of SCNN1G in a Chinese Family.

Objective: Liddle syndrome (LS) is a monogenic hypertension consistent with autosomal dominant inheritance, often with early onset high blood pressure in childhood or adolescence. This study aimed to identify the pathogenicity of a nonsense mutation in SCNN1G in a Chinese family with LS and the long-term outcomes of tailored treatment with amiloride. Methods: To explore the pathogenicity of candidate variant reported in 2015 by our team, we constructed mutant and wild-type models in vitro and measured amiloride-sensitive current in Chinese Hamster Ovary (CHO) cells using patch clamp technique. Participants were followed up for 7 years after tailored treatment with amiloride. Results: A nonsense variant was detected in six members, two of whom were pediatric patients. This mutation resulted in a termination codon at codon 572, truncating the Pro-Pro-Pro-X-Tyr motif. The mutant epithelial sodium channels displayed higher amiloride-sensitive currents than the wild-type channels (P < 0.05). Tailored treatment with amiloride achieved ideal blood pressure control in all patients with normal cardiorenal function, and no adverse events occurred during follow-up. Conclusion: We found the pathogenicity of a nonsense SCNN1G mutation (p.Glu571*) with enhanced amiloride-sensitive currents in a LS family with young patients. Tailored treatment with amiloride may be an effective strategy for the long-term control of blood pressure and protection from target organ damage or cardiovascular events, including children and youth patients with LS.

Implication of a novel truncating mutation in titin as a cause of autosomal dominant left ventricular noncompaction.

BACKGROUND: Mutation in the titin gene (TTN) in left ventricular noncompaction (LVNC) has been reported with a highly heterogeneous prevalence, and the molecular mechanisms underlying the pathogenesis of TTN gene mutation are uncharacterized. In the present study, we identified a novel TTN mutation in a pedigree with LVNC and investigated the potential pathogenic mechanism by functional studies. METHODS: The whole-genome sequencing with linkage analysis was performed in a 3-generation family affected by autosomal dominant LVNC cardiomyopathy. The clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR/Cas9) technology was used to establish novel truncating mutation in TTN in a rat cardiomyoblast H9C2 cell line in vitro, in which functional studies were carried out and characterized in comparison to its wild-type counterpart. RESULTS: A novel truncating mutation TTN p. R2021X was identified as the only plausible disease-causing variant that segregated with disease among the five surviving affected individuals, with an interrogation of the entire genome excluding other potential causes. Quantitative reverse transcription-polymerase chain reaction and cellular immunofluorescence supported a haploinsufficient disease mechanism in titin truncation mutation cardiomyocytes. Further functional studies suggested mitochondrial abnormities in the presence of mutation, including decreased oxygen consumption rate, reduced adenosine triphosphate production, impaired activity of electron translation chain, and abnormal mitochondrial structure on electron microscopy. Impaired autophagy under electron microscopy accompanied with activation of the Akt-mTORC1 signaling pathway was observed in TTN p. R2021X truncation mutation cardiomyocytes. CONCLUSIONS: The TTN p. R2021X mutation has a function in the cause of a highly penetrant familial LVNC. These findings expand the spectrum of titin's roles in cardiomyopathies and provide novel insight into the molecular basis of titin-truncating variants-associated LVNC.