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Previously, we demonstrated that mitochondrial transplantation has beneficial effects in a polymicrobial sepsis model. However, the mechanism has not been fully investigated. Mitochondria have their own genes, and genomic changes in sepsis are an important issue in terms of pathophysiology, biomarkers, and therapeutic targets. To investigate the changes in transcriptomic features after mitochondrial transplantation in a polymicrobial sepsis model, we used a rat model of fecal slurry polymicrobial sepsis. Total RNA from splenocytes of sham-operated (SHAM, n = 10), sepsis-induced (SEPSIS, n = 7), and sepsis receiving mitochondrial transplantation (SEPSIS + MT, n = 8) samples was extracted and we conducted a comparative transcriptome-wide analysis between three groups. We also confirmed these results with qPCR. In terms of percentage of mitochondrial mapped reads, the SEPSIS + MT group had a significantly higher mapping ratio than the others. RT1-M2 and Cbln2 were identified as highly expressed in SEPSIS + MT compared with SEPSIS. Using SHAM expression levels as another control variable, we further identified six genes (Fxyd4, Apex2l1, Kctd4, 7SK, SNORD94, and SNORA53) that were highly expressed after sepsis induction and observed that their expression levels were attenuated by mitochondrial transplantation. Changes in transcriptomic features were identified after mitochondrial transplantation in sepsis. This might provide a hint for exploring the mechanism of mitochondrial transplantation in sepsis.
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Sepsis , Transcriptoma , Ratas , Animales , Mitocondrias/genética , Mitocondrias/metabolismo , Perfilación de la Expresión Génica , Sepsis/genética , Sepsis/metabolismoRESUMEN
Previously, we have shown that mitochondrial transplantation in the sepsis model has immune modulatory effects. The mitochondrial function could have different characteristics dependent on cell types. Here, we investigated whether the effects of mitochondrial transplantation on the sepsis model could be different depending on the cell type, from which mitochondria were isolated. We isolated mitochondria from L6 muscle cells, clone 9 liver cells and mesenchymal stem cells (MSC). We tested the effects of mitochondrial transplantation using in vitro and in vivo sepsis models. We used the LPS stimulation of THP-1 cell, a monocyte cell line, as an in vitro model. First, we observed changes in mitochondrial function in the mitochondria-transplanted cells. Second, we compared the anti-inflammatory effects of mitochondrial transplantation. Third, we investigated the immune-enhancing effects using the endotoxin tolerance model. In the in vivo polymicrobial fecal slurry sepsis model, we examined the survival and biochemical effects of each type of mitochondrial transplantation. In the in vitro LPS model, mitochondrial transplantation with each cell type improved mitochondrial function, as measured by oxygen consumption. Among the three cell types, L6-mitochondrial transplantation significantly enhanced mitochondrial function. Mitochondrial transplantation with each cell type reduced hyper-inflammation in the acute phase of in vitro LPS model. It also enhanced immune function during the late immune suppression phase, as shown by endotoxin tolerance. These functions were not significantly different between the three cell types of origin for mitochondrial transplantation. However, only L6-mitochondrial transplantation significantly improved survival compared to the control in the polymicrobial intraabdominal sepsis model. The effects of mitochondria transplantation on both in vitro and in vivo sepsis models differed depending on the cell types of origin for mitochondria. L6-mitochondrial transplantation might be more beneficial in the sepsis model.
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Lipopolisacáridos , Sepsis , Humanos , Lipopolisacáridos/metabolismo , Mitocondrias/metabolismo , Sepsis/metabolismo , Inflamación/metabolismo , Monocitos/metabolismoRESUMEN
Hepatocellular carcinoma (HCC) is the second most common cancer worldwide, demonstrating aggressiveness and mortality more frequently in men than in women. Despite reports regarding the inhibitory ability of estrogen receptor alpha (ERα, ESR1) in certain cancer progression, targets and the basis of underlying gender disparity in HCC worsening remain elusive. Here, we report the ability of ERα to transcriptionally inhibit G protein subunit alpha 12 (Gα12) responsible for HCC worsening. First, using human samples and public database, the expression of ERα and Gα12 in HCC was examined. Then, quantitative real-time PCR, chromatin immunoprecipitation-assay, luciferase assay and immunoblottings of liver cancer cell lines confirmed the inhibitory ability of ERα on Gα12 and HCC progression. Gα12 promoted mesenchymal characteristics and amoeboidal movement, which was antagonized by ERα overexpression. Additionally, we found microRNA-141 and microRNA-200a as downstream targets of the Gα12 signaling axis for cancer malignancy regulation under the control of ERα. As for in-depth mechanism, PTP4A1 was found to be directly inhibited by microRNA-141 and microRNA-200a. Moreover, we found the inhibitory effect of ERα on amoeboidal movement by analyzing the morphology and blebbing of liver cancer cells and the active form of MLC levels. The identified targets and ESR1 levels are inversely correlated with human specimens, as well as with sex-biased survival rates of HCC patients. Collectively, ERα-dependent repression of Gα12 and consequent changes in the Gα12 signaling may explain the gender disparity in HCC, providing pharmacological clues for the control of metastatic HCC.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , MicroARNs , Carcinoma Hepatocelular/patología , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Femenino , Subunidades alfa de la Proteína de Unión al GTP G12-G13/genética , Subunidades alfa de la Proteína de Unión al GTP G12-G13/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Hepáticas/patología , Masculino , MicroARNs/metabolismoRESUMEN
BACKGROUND AND AIMS: Fat accumulation results from increased fat absorption and/or defective fat metabolism. Currently, the lipid-sensing nuclear receptor that controls fat utilization in hepatocytes is elusive. Liver X receptor alpha (LXRα) promotes accumulation of lipids through the induction of several lipogenic genes. However, its effect on lipid degradation is open for study. Here, we investigated the inhibitory role of LXRα in autophagy/lipophagy in hepatocytes and the underlying basis. APPROACH AND RESULTS: In LXRα knockout mice fed a high-fat diet, or cell models, LXRα activation suppressed the function of mitochondria by inhibiting autophagy/lipophagy and induced hepatic steatosis. Gene sets associated with "autophagy" were enriched in hepatic transcriptome data. Autophagy flux was markedly augmented in the LXRα knockout mouse liver and primary hepatocytes. Mechanistically, LXRα suppressed autophagy-related 4B cysteine peptidase (ATG4B) and Rab-8B, responsible for autophagosome and -lysosome formation, by inducing let-7a and microRNA (miR)-34a. Chromatin immunoprecipitation assay enabled us to find LXRα as a transcription factor of let-7a and miR-34a. Moreover, 3' untranslated region luciferase assay substantiated the direct inhibitory effects of let-7a and miR-34a on ATG4B and Rab-8B. Consistently, either LXRα activation or the let-7a/miR-34a transfection lowered mitochondrial oxygen consumption rate and mitochondrial transmembrane potential and increased fat levels. In obese animals or nonalcoholic fatty liver disease (NAFLD) patients, let-7a and miR-34a levels were elevated with simultaneous decreases in ATG4B and Rab-8B levels. CONCLUSIONS: LXRα inhibits autophagy in hepatocytes through down-regulating ATG4B and Rab-8B by transcriptionally activating microRNA let-7a-2 and microRNA 34a genes and suppresses mitochondrial biogenesis and fuel consumption. This highlights a function of LXRα that culminates in the progression of liver steatosis and steatohepatitis, and the identified targets may be applied for a therapeutic strategy in the treatment of NAFLD.
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Proteínas Relacionadas con la Autofagia/metabolismo , Autofagia/fisiología , Cisteína Endopeptidasas/metabolismo , Hepatocitos/metabolismo , Receptores X del Hígado/metabolismo , Mitocondrias/fisiología , Proteínas de Unión al GTP rab/metabolismo , Activación Metabólica , Animales , Autofagia/genética , Proteínas Relacionadas con la Autofagia/genética , Cisteína Endopeptidasas/genética , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Regulación hacia Abajo , Hígado Graso/etiología , Hígado Graso/genética , Hígado Graso/metabolismo , Hígado Graso/fisiopatología , Células Hep G2/metabolismo , Células Hep G2/fisiología , Hepatocitos/fisiología , Humanos , Metabolismo de los Lípidos/genética , Metabolismo de los Lípidos/fisiología , Hígado/metabolismo , Hígado/fisiología , Hígado/fisiopatología , Receptores X del Hígado/genética , Receptores X del Hígado/fisiología , Ratones , Ratones Noqueados , MicroARNs/genética , MicroARNs/metabolismo , MicroARNs/fisiología , Mitocondrias/metabolismo , Enfermedad del Hígado Graso no Alcohólico/etiología , Enfermedad del Hígado Graso no Alcohólico/genética , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Enfermedad del Hígado Graso no Alcohólico/fisiopatología , Biogénesis de Organelos , Consumo de Oxígeno/genética , Consumo de Oxígeno/fisiología , Transcriptoma , Proteínas de Unión al GTP rab/genéticaRESUMEN
Immune suppression is known to occur during sepsis. Endotoxin tolerance is considered a mechanism of immune suppression in sepsis. However, the timing and serial changes in endotoxin tolerance have not been fully investigated. In this study, we investigated serial changes in endotoxin tolerance in a polymicrobial sepsis model. Herein, we used a rat model of fecal slurry polymicrobial sepsis. After induction of sepsis, endotoxin tolerance of peripheral blood mononuclear cells (PBMCs) and splenocytes was measured at various time points (6 h, 12 h, 24 h, 48 h, 72 h, 5 days, and 7 days), through the measurement of TNF-α production after stimulation with lipopolysaccharide (LPS) in an ex vivo model. At each time point, we checked for plasma tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10 levels. Moreover, we analyzed reactive oxygen species (ROS) as measured by 2',7'-dichlorodihydrofluorescein, plasma lactate, serum alanine aminotransferase (ALT), and creatinine levels. Nuclear factor (NF)-κB, IL-1 receptor-associated kinase (IRAK)-M, and cleaved caspase 3 levels were measured in the spleen. Endotoxin tolerance, measured by TNF-α production stimulated through LPS in PBMCs and splenocytes, was induced early in the sepsis model, starting from 6 h after sepsis. It reached a nadir at 24 to 48 h after sepsis, and then started to recover. Endotoxin tolerance was more prominent in the severe sepsis model. Plasma cytokines peaked at time points ranging from 6 to 12 h after sepsis. ROS levels peaked at 12 h and then decreased. Lactate, ALT, and serum creatinine levels increased up to 24 to 48 h, and then decreased. Phosphorylated p65 and IRAK-M levels of spleen increased up to 12 to 24 h and then decreased. Apoptosis was prominent 48 h after sepsis, and then recovered. In the rat model of polymicrobial sepsis, endotoxin tolerance occurred earlier and started to recover from 24 to 48 h after sepsis.
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Lipopolisacáridos , Sepsis , Animales , Tolerancia a Endotoxinas , Interleucina-6 , Lactatos , Leucocitos Mononucleares , Lipopolisacáridos/farmacología , FN-kappa B , Ratas , Especies Reactivas de Oxígeno , Sepsis/patología , Factor de Necrosis Tumoral alfaRESUMEN
OBJECTIVE: Takotsubo syndrome following cardiac surgery is a rare occurrence. However, early diagnosis is essential to prevent treatment which could increase the left ventricular outflow tract pressure gradient in patients with Takotsubo syndrome, and lead to cardiogenic shock. Therefore, our study aimed to identify the incidence of Takotsubo syndrome after cardiopulmonary bypass and the associated risk factors and prognosis. METHODS: We retrospectively studied 5773 patients who underwent cardiopulmonary bypass between February 2007 and July 2017. Among these, Takotsubo syndrome was diagnosed in 52 (0.9%). To evaluate the risk factors for Takotsubo syndrome, 104 of the remaining 5721 patient were randomly selected as the control group (1:2 ratio). Univariate and multivariate logistic regression analyses were used for risk factor analysis. RESULTS: Majority of patients (69.2%) in the Takotsubo syndrome group underwent mitral valve surgery, compared with 32.7% in the control group. The following risk factors of Takotsubo syndrome were identified: atrio-ventricular valve surgery (odds ratio (OR) 10.5; 95% confidence interval (CI), 2.6-42.5; p = 0.001); and the immediate postoperative use of epinephrine (OR, 3.3; 95% CI, 1.0-10.7; p = 0.05) and dobutamine (OR, 4.8; 95% CI, 1.72-13.3; p = 0.003). Hypertension was a significant protective factor against Takotsubo syndrome following cardiac surgery (OR, 0.22; 95% CI, 0.06-0.73; p = 0.01). CONCLUSION: Takotsubo syndrome following cardiac surgery is rare. Immediate postoperative use of epinephrine and doputamine, as well as atrio-ventricular valve surgery were factors associated with the development of Takotsubo syndrome.
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Procedimientos Quirúrgicos Cardíacos , Cardiomiopatía de Takotsubo , Procedimientos Quirúrgicos Cardíacos/efectos adversos , Estudios de Casos y Controles , Humanos , Estudios Retrospectivos , Factores de Riesgo , Cardiomiopatía de Takotsubo/epidemiología , Cardiomiopatía de Takotsubo/etiologíaRESUMEN
Colour is one of the important quality traits affecting the meat purchasing decision by consumers, and myoglobin is the principal heme protein responsible for the meat colour. This study aimed to investigate the effects of pH (5.3, 5.8, 6.4 and 7.4) and temperature (4 and 25 °C) on oxymyoglobin (OxyMb) oxidation in model reaction mixtures containing OxyMb, fatty acids (C18:2n-6 and C18:3n-3) and vitamin E. A decrease of the OxyMb concentration with increased acidity was observed for all the reaction mixtures with/without fatty acids and vitamin E. After 48 h of storage at 4 °C, the OxyMb concentration decreased by approximately 60-70%, 61-69%, 53.7-53.9% and 40.93-41.84% in the reaction mixtures containing [OxyMb + C18:2n-6 or C18:3n-3] at pH 5.3, 5.8, 6.4 and 7.4, respectively. While, after 48 h at 25 °C, the OxyMb concentration decreased by 95-98% in all the reaction mixtures containing [OxyMb + C18:2n-6 or C18:3n-3] under all the pH conditions. The presence of vitamin E significantly inhibited the OxyMb oxidation in the reaction mixtures containing fatty acids under acidic conditions, but a higher level of vitamin E may be required for meat(s) containing high n-3 fatty acids content that are stored at high temperature.
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Activation of sterol regulatory element-binding protein 1 (SREBP-1), a master lipogenic transcription factor, is associated with cancer metabolism and metabolic disorders. Neddylation, the process of adding NEDD8 to its substrate, contributes to diverse biological processes. Here, we identified SREBP-1 as a substrate for neddylation by UBC12 and explored its impact on tumor aggressiveness. In cell-based assays, SREBP-1 neddylation prolonged SREBP-1 stability with a decrease in ubiquitination. Consequently, NEDD8 overexpression facilitated proliferation, migration, and invasion of SK-Hep1 liver tumor cells. MLN4924 (an inhibitor of the NEDD8-activating enzyme-E1) treatment or UBC12 knockdown prevented SREBP-1 neddylation and tumor cell phenotype change. This effect was corroborated in an in vivo xenograft model. In human specimens, SREBP-1, UBC12, and NEDD8 were all upregulated in hepatocellular carcinoma (HCC) compared to nontumorous regions. Moreover, SREBP-1 levels positively correlated with UBC12. In GEO database analyses, SREBP-1 levels were greater in metastatic HCC samples accompanying UBC12 upregulation. In HCC analysis, tumoral SREBP-1 and UBC12 levels discriminated overall patient survival rates. Additionally, MLN4924 treatment destabilized SREBP-1 in MDA-MB-231 breast cancer cells and in the tumor cell xenograft. SREBP-1 and UBC12 were also highly expressed in human breast cancer tissues. Moreover, most breast cancers with lymph node metastasis displayed predominant SREBP-1 and UBC12 expressions, which compromised overall patient survival rates. In summary, SREBP-1 is neddylated by UBC12, which may contribute to HCC and breast cancer aggressiveness through SREBP-1 stabilization, and these events can be intervented by MLN4924 therapy. Our findings may also provide potential reliable prognostic markers for tumor metastasis.
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Neoplasias de la Mama/mortalidad , Carcinoma Hepatocelular/mortalidad , Neoplasias Hepáticas/mortalidad , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo , Enzimas Ubiquitina-Conjugadoras/metabolismo , Animales , Mama/patología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/secundario , Línea Celular Tumoral , Ciclopentanos/farmacología , Ciclopentanos/uso terapéutico , Femenino , Humanos , Hígado/patología , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/patología , Metástasis Linfática/patología , Ratones , Proteína NEDD8/metabolismo , Pronóstico , Estabilidad Proteica/efectos de los fármacos , Pirimidinas/farmacología , Pirimidinas/uso terapéutico , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/análisis , Tasa de Supervivencia , Enzimas Activadoras de Ubiquitina/antagonistas & inhibidores , Enzimas Activadoras de Ubiquitina/metabolismo , Enzimas Ubiquitina-Conjugadoras/análisis , Ubiquitinación/efectos de los fármacos , Regulación hacia Arriba , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
OBJECTIVE: Although the slaughter of cull sows (CS) for human consumption and meat products processing appears quite common throughout the world, relatively limited scientific information regarding the meat quality parameters of this pork type is available. The present study aimed at providing the technological quality characteristics and eating quality of CS meat, and comparing with those of commercial pork. METHODS: Longissimus thoracis et lumborum muscle samples of CS and finisher pigs (FP) at 24 h postmortem were collected and used for investigation of the meat quality traits (pH, color, shear force, cooking loss, water holding capacity), fatty acids, flavor compounds and sensory characteristics. RESULTS: The CS meat had significantly higher moisture content (p = 0.0312) and water holding capacity (p = 0.0213) together with lower cooking loss (p = 0.0366) compared to the FP meat. The CS meat also exhibited higher (p = 0.0409) contents of unsaturated fatty acids, especially polyunsaturated fatty acids (PUFA, p = 0.0213) and more desirable PUFA/total saturated fatty acids ratio (p = 0.0438) compared to the FP meat. A total of 56 flavor compounds were identified, amongst the amount of 16 compounds differed significantly between the two pork groups. Most of the PUFA-derived flavor compounds (e.g., hexanal, benzaldehyde, and hydrocarbons) showed higher amounts in the CS meat. While, 3-(methylthio)-propanal and 4-methylthiazole associated with pleasant aromas (meaty and roast odor notes) were only found in the FP meat. Furthermore, no differences were reported by panelists for flavor, juiciness, tenderness, and acceptability scores between the two pork groups studied. CONCLUSION: The sow meat exhibited better technological quality and its eating quality could be comparable to the commercial pork. This study provides meat processors and traders with valuably scientific information which may help to improve the utilization and consumption level of sow meat.
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Objective: The present work aimed at evaluating the effects of carcass quality grade on the quality characteristics of pork meat according to Korean carcass quality grade system. Method: Pork carcasses with varying in quality grades (QG): 1+ (QG1+, n=10), 1 (QG1, n=10) and 2 (QG2, n=10), were used to evaluate the relationship between carcass quality grade and meat quality. The meat quality traits, fatty acid profiles, flavor compounds and sensory qualities were measured on the longissimus dorsi muscle samples of these carcasses. Results: Pork meat of higher quality grade (GQ1+) presented significantly higher fat content (5.43%), C18:2n-6 level (19.03%) and total unsaturated fatty acids (UFA) content (62.72%). Also, the QG1+ meat was significantly higher in levels of classes of flavor compounds such as aldehydes, alcohols and hydrocarbons in comparison to those of the meat samples from the lower GQ groups. The sensory evaluation results (flavor, juiciness, tenderness and acceptability scores) of QG1+ meat was significantly higher than the QG1 and QG2 meats. The pork with lower QG (i.e., QG2) was found positively correlated to redness (r=0.987), C18:1n-9 level (r=1.000) but negatively correlated to the fat content (r=-0.949), and flavor (r=-0.870), juiciness (r=-0.861), tenderness (r=-0.862) and acceptability (r=-0.815) scores. Conclusion: The pork with higher quality grade had higher fat content, total unsaturated fatty acids and better eating quality, thus producing pork with higher quality grades should be considered in order to satisfy the consumer's expectation.
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BACKGROUND & AIMS: Hepatic stellate cells (HSCs) have a role in liver fibrosis. Guanine nucleotide-binding α-subunit 12 (Gα12) converges signals from G-protein-coupled receptors whose ligand levels are elevated in the environment during liver fibrosis; however, information is lacking on the effect of Gα12 on HSC trans-differentiation. This study investigated the expression of Gα12 in HSCs and the molecular basis of the effects of its expression on liver fibrosis. METHODS: Gα12 expression was assessed by immunostaining, and immunoblot analyses of mouse fibrotic liver tissues and primary HSCs. The role of Gα12 in liver fibrosis was estimated using a toxicant injury mouse model with Gα12 gene knockout and/or HSC-specific Gα12 delivery using lentiviral vectors, in addition to primary HSCs and LX-2 cells using microRNA (miR) inhibitors, overexpression vectors, or adenoviruses. miR-16, Gα12, and LC3 were also examined in samples from patients with fibrosis. RESULTS: Gα12 was overexpressed in activated HSCs and fibrotic liver, and was colocalised with desmin. In a carbon tetrachloride-induced fibrosis mouse model, Gα12 ablation prevented increases in fibrosis and liver injury. This effect was attenuated by HSC-specific lentiviral delivery of Gα12. Moreover, Gα12 activation promoted autophagy accompanying c-Jun N-terminal kinase-dependent ATG12-5 conjugation. In addition, miR-16 was found to be a direct inhibitor of the de novo synthesis of Gα12. Modulations of miR-16 altered autophagy in HSCs. In a fibrosis animal model or patients with severe fibrosis, miR-16 levels were lower than in their corresponding controls. Consistently, cirrhotic patient liver tissues showed Gα12 and LC3 upregulation in desmin-positive areas. CONCLUSIONS: miR-16 dysregulation in HSCs results in Gα12 overexpression, which activates HSCs by facilitating autophagy through ATG12-5 formation. This suggests that Gα12 and its regulatory molecules could serve as targets for the amelioration of liver fibrosis. LAY SUMMARY: Guanine nucleotide-binding α-subunit 12 (Gα12) is upregulated in activated hepatic stellate cells (HSCs) as a consequence of the dysregulation of a specific microRNA that is abundant in HSCs, facilitating the progression of liver fibrosis. This event is mediated by c-Jun N-terminal kinase-dependent ATG12-5 formation and the promotion of autophagy. We suggest that Gα12 and its associated regulators could serve as new targets in HSCs for the treatment of liver fibrosis.
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Subunidades alfa de la Proteína de Unión al GTP G12-G13/metabolismo , Células Estrelladas Hepáticas/metabolismo , Cirrosis Hepática , MicroARNs/metabolismo , Animales , Autofagia/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Subunidades alfa de la Proteína de Unión al GTP G12-G13/antagonistas & inhibidores , Regulación de la Expresión Génica , Humanos , Cirrosis Hepática/metabolismo , Cirrosis Hepática/patología , Ratones , Inhibidor 1 de Activador Plasminogénico/metabolismo , Inhibidor 1 de Activador Plasminogénico/farmacología , Inhibidores de Serina Proteinasa/farmacología , Transducción de Señal/efectos de los fármacos , Regulación hacia ArribaRESUMEN
A Bacillus species that produces 2,3-butanediol (2,3-BD), termed BRC1, was newly isolated, and a 2,3-BD dehydrogenase (Bdh) from this species was identified and characterized at the molecular and biochemical level. Sequence analysis revealed that Bdh is homologous to D-2,3-BD dehydrogenases. An analysis of the enzymatic properties of Bdh overexpressed in Escherichia coli confirmed the molecular results, showing preferred activity toward D-2,3-BD. Optimum pH, temperature, and kinetics determined for reductive and oxidative reactions support the preferential production of 2,3-BD during cell growth. Overexpression of bdh under the control of a xylose-inducible promoter resulted in increased enzyme activity and enhanced 2,3-BD production in Bacillus sp. BRC1. Additionally, a hydrolysate of cellulosic material, (empty palm fruit bunches), was successfully used for the enhanced production of 2,3-BD in the recombinant Bacillus strain.
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Oxidorreductasas de Alcohol/metabolismo , Arecaceae/microbiología , Bacillus/fisiología , Butileno Glicoles/aislamiento & purificación , Butileno Glicoles/metabolismo , Frutas/microbiología , Oxidorreductasas de Alcohol/genética , Bacillus/clasificación , Mejoramiento Genético/métodos , Hidrólisis , Especificidad de la EspecieRESUMEN
A 62-year-old woman who had undergone mitral valve replacement 24 years ago was admitted to the hospital with congestive heart failure. She needed heart transplantation for stage D heart failure. Preoperative cardiac computed tomographic scans showed a severely calcified left atrium and a large right atrium. Given that the left atrium's calcification was too severe to suture, the calcified left atrial wall was broadly resected, and the resected left atrial wall was reconstructed with a bovine pericardial patch for anastomosis with the donor's left atrial wall. The operation was completed without heavy bleeding, and the patient was discharged from the hospital with no complications.
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Calcinosis , Atrios Cardíacos , Insuficiencia Cardíaca , Trasplante de Corazón , Cardiopatía Reumática , Tomografía Computarizada por Rayos X , Humanos , Femenino , Cardiopatía Reumática/cirugía , Cardiopatía Reumática/complicaciones , Cardiopatía Reumática/diagnóstico , Trasplante de Corazón/métodos , Persona de Mediana Edad , Calcinosis/cirugía , Calcinosis/diagnóstico , Calcinosis/complicaciones , Atrios Cardíacos/diagnóstico por imagen , Atrios Cardíacos/cirugía , Insuficiencia Cardíaca/cirugía , Insuficiencia Cardíaca/etiología , Insuficiencia Cardíaca/diagnóstico , Índice de Severidad de la Enfermedad , Resultado del Tratamiento , Implantación de Prótesis de Válvulas Cardíacas/métodos , Pericardio/trasplante , Pericardio/cirugíaRESUMEN
Endoplasmic reticulum (ER) stress plays a crucial role in liver diseases, affecting various types of hepatic cells. While studies have focused on the link between ER stress and hepatocytes as well as hepatic stellate cells (HSCs), the precise involvement of hepatic macrophages in ER stress-induced liver injury remains poorly understood. Here, we examined the effects of ER stress on hepatic macrophages and their role in liver injury. Acute ER stress led to the accumulation and activation of hepatic macrophages, which preceded hepatocyte apoptosis. Notably, macrophage depletion mitigated liver injury induced by ER stress, underscoring their detrimental role. Mechanistic studies revealed that ER stress stimulates macrophages predominantly via the PERK signaling pathway, regardless of its canonical substrate ATF4. hnRNPA1 has been identified as a crucial mediator of PERK-driven macrophage activation, as the overexpression of hnRNPA1 effectively reduced ER stress and suppressed pro-inflammatory activation. We observed that hnRNPA1 interacts with mRNAs that encode UPR-related proteins, indicating its role in the regulation of ER stress response in macrophages. These findings illuminate the cell type-specific responses to ER stress and the significance of hepatic macrophages in ER stress-induced liver injury. Collectively, the PERK-hnRNPA1 axis has been discovered as a molecular mechanism for macrophage activation, presenting prospective therapeutic targets for inflammatory hepatic diseases such as acute liver injury.
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BACKGROUND: Atrial functional mitral regurgitation (AFMR) is a newly discovered condition associated with longstanding atrial fibrillation. This retrospective study aimed to analyze the outcomes of the maze procedure and mitral regurgitation (MR) surgery in AFMR and atrial fibrillation in comparison with those in degenerative MR (DMR). METHODS: Patients who underwent mitral valve repair/replacement with a maze procedure at a hospital (July 2012-August 2021) were included. We excluded patients aged below 18 years undergoing concomitant coronary artery bypass grafting or atrial septal defect repair and those with MR etiology other than ARMR or DMR. RESULTS: We included 35 patients with AFMR and 50 patients with DMR. Patient characteristics and postoperative outcomes were not significantly different between the two groups. Long-term outcomes revealed no significant differences in the ratio of cardiac mortality, stroke, or hospital readmission. However, after the maze procedure, the sinus rhythm restoration rate was significantly lower (62% vs. 28.5%, p < 0.001), a junctional rhythm state (p < 0.001) and permanent pacemaker insertion for sick sinus syndrome (SSS) (p = 0.03) were significantly more common in AFMR than DMR. On postoperative transthoracic echocardiography (TTE), the pulmonary artery systolic pressure was significantly less decreased in the AFMR group than in the DMR group compared with that on preoperative TTE (p = 0.04). CONCLUSIONS: AFMR showed excellent mitral valve surgery outcomes, similar to DMR, but had a significantly higher risk of pacemaker insertion for SSS after the maze procedure.
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Fibrilación Atrial , Procedimiento de Laberinto , Insuficiencia de la Válvula Mitral , Válvula Mitral , Humanos , Insuficiencia de la Válvula Mitral/cirugía , Insuficiencia de la Válvula Mitral/fisiopatología , Masculino , Estudios Retrospectivos , Femenino , Persona de Mediana Edad , Fibrilación Atrial/cirugía , Fibrilación Atrial/fisiopatología , Resultado del Tratamiento , Válvula Mitral/cirugía , Válvula Mitral/fisiopatología , Anciano , Implantación de Prótesis de Válvulas Cardíacas/métodos , Ecocardiografía , Complicaciones Posoperatorias/fisiopatología , Complicaciones Posoperatorias/epidemiologíaRESUMEN
Background: Pump-controlled retrograde trial off (PCRTO) is a safe, simple, and reversible method for weaning patients from veno-arterial extracorporeal membrane oxygenation (VA-ECMO). However, few studies have compared PCRTO to conventional weaning methods. This retrospective study aimed to compare PCRTO to non-PCRTO methods. Methods: This study included patients who were weaned from VA-ECMO from January 2016 to December 2022 at our medical center. Demographic data, ECMO management, ECMO complications, survival to discharge, and cardiogenic shock after VA-ECMO weaning were compared between the 2 groups. Results: Seventy patients who were weaned from VA-ECMO using PCRTO and 85 patients who were weaned with conventional methods were compared. Patient characteristics were not significantly different between the 2 groups. The rate of survival to discharge was significantly higher in the PCRTO group than in the non-PCRTO group (90% vs. 72%, p=0.01). The rates of freedom from all-cause mortality at 10, 30, and 50 days after weaning from ECMO were 75%, 55%, and 35% in the non-PCRTO group and 62%, 60%, and 58% in the PCRTO group, respectively (p=0.1). The incidence of cardiogenic shock after weaning from VA-ECMO was significantly higher in the non-PCRTO group (16% vs. 5%, p=0.04). In logistic regression analysis, PCRTO was a significant factor for survival to discharge (odds ratio, 2.42; 95% confidence interval, 1.29-5.28; p=0.02). Conclusion: Compared to conventional methods, PCRTO is a feasible and reversible method, and it serves as a useful predictor of successful VA-ECMO weaning through a preload stress test.
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OBJECTIVE: Beef of Jeju black cattle (JBC) is considered as a healthy meat type due to its significantly higher unsaturated fatty acids (UFA). Lipid (e.g., UFA) is highly susceptible to oxidizing agents, which results in the quality deterioration and economic value loss of meat products. Therefore, development and application of novel preservative techniques is necessary to improve the shelf-life stability of high-UFA beef. The objective of this study was to assess the applicability of chitosan-based coatings in preservation of JBC beef. METHODS: Different coating solutions: 2% chitosan alone, and 2% chitosan containing 0.1% or 0.3% gallic acid were prepared to investigate their applicability in preservation of fresh beef during storage. Jeju black cattle beef (2-cm thick steaks) were non-coated (control) or coated with the above coating solutions, placed on trays, over-wrapped with plastic film and stored at 4°C. The microbiological indices, color, total volatile basic nitrogen (TVBN) and lipid oxidation of the beef were investigated after 1, 10, and 21 days of storage. RESULTS: Coating with 2% chitosan alone reduced the spoilage bacteria count, TVBN and thiobarbituric acid reactive substances levels in the beef compared with control during storage (p<0.05). Noticeably, coating with 2% chitosan containing 0.1% or 0.3% gallic acid was more effective on retardation of spoilage bacteria growth, lipid oxidation and discoloration in the beef compared to the chitosan coating alone over the storage period (21 days) (p<0.05). CONCLUSION: Taken together, the combined chitosan and gallic acid coating could be used as a bio-preservative technique in the meat industry.
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Rationale: The surge of severe liver damage underscores the necessity for identifying new targets and therapeutic agents. Endoplasmic reticulum (ER) stress induces ferroptosis with Gα12 overexpression. NF-κB essential modulator (NEMO) is a regulator of inflammation and necroptosis. Nonetheless, the regulatory basis of NEMO de novo synthesis and its impact on hepatocyte ferroptosis need to be established. This study investigated whether Nrf2 transcriptionally induces IKBKG (the NEMO gene) for ferroptosis inhibition and, if so, how NEMO induction protects hepatocytes against ER stress-induced ferroptosis. Methods: Experiments were conducted using human liver tissues, hepatocytes, and injury models, incorporating NEMO overexpression and Gα12 gene modulations. RNA sequencing, immunoblotting, immunohistochemistry, reporter assays, and mutation analyses were done. Results: NEMO downregulation connects closely to ER and oxidative stress, worsening liver damage via hepatocyte ferroptosis. NEMO overexpression protects hepatocytes from ferroptosis by promoting glutathione peroxidase 4 (GPX4) expression. This protective role extends to oxidative and ER stress. Similar shifts occur in nuclear factor erythroid-2-related factor-2 (Nrf2) expression alongside NEMO changes. Nrf2 is newly identified as an IKBKG (NEMO gene) transactivator. Gα12 changes, apart from Nrf2, impact NEMO expression, pointing to post-transcriptional control. Gα12 reduction lowers miR-125a, an inhibitor of NEMO, while overexpression has the opposite effect. NEMO also counters ER stress, which triggers Gα12 overexpression. Gα12's significance in NEMO-dependent hepatocyte survival is confirmed via ROCK1 inhibition, a Gα12 downstream kinase, and miR-125a. The verified alterations or associations within the targeted entities are validated in human liver specimens and datasets originating from livers subjected to exposure to other injurious agents. Conclusions: Hepatic injury prompted by ER stress leads to the suppression of NEMO, thereby facilitating ferroptosis through the inhibition of GPX4. IKBKG is transactivated by Nrf2 against Gα12 overexpression responsible for the increase of miR-125a, an unprecedented NEMO inhibitor, resulting in GPX4 induction. Accordingly, the induction of NEMO mitigates ferroptotic liver injury.
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Ferroptosis , Hepatopatías , MicroARNs , Humanos , Estrés del Retículo Endoplásmico/genética , Ferroptosis/genética , Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , MicroARNs/genética , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , FN-kappa B/metabolismo , Quinasas Asociadas a rhoRESUMEN
This study was carried out to assess the quality properties, components associated with taste and aroma of beef as a function of breed. For this purpose, steers from four Korean native cattle breeds: Hanwoo (n=10), Chikso (n=10), black Hanwoo (n=12, BHW) and Jeju black cattle (n=12, JBC) were used. The steers all were raised under identical conditions and finished at a similar age of around 30-months old. Following 24 h of slaughter, all longissimus lumborum muscles were collected and used for analysis of meat quality, fatty acids, and flavor-related components (metabolic compounds, free amino acids, and aroma volatiles). The Hanwoo presented a significantly higher intramuscular fat content (IMF, 22.85%) than the BHW (11.78%), Chikso (9.25%), and JBC (9.14%; p<0.05). The meat of Hanwoo breed showed lighter and redder color, and lower shear force value (p<0.05). The JBC presented a "healthier" fatty acid profiles as it had a higher total unsaturated fatty acids content (p<0.05). With regard to flavor-related components, Hanwoo also had higher total contents of free amino acids and metabolites associated with umami and sweet tastes, and fat-derived volatile compounds (aldehydes, alcohols, and ketones) associated with fatty aroma. It may be concluded that there was a considerable difference in the meat quality properties among breeds. The variations of IMF content and flavor-related components may be the main factors contributing to the typical flavors of beef among the four Korean native cattle breeds.
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OBJECTIVE: To analyze and compare the outcomes of mitral valve surgery for atrial functional mitral regurgitation (AFMR) and for degenerative mitral regurgitation (DMR). METHODS: Patients with AFMR or DMR who underwent mitral valve repair/replacement at 2 institutions between January 2012 and December 2022 were included. Patients <18 years of age and patients undergoing concomitant cardiac surgery (except for the maze procedure or tricuspid annuloplasty) were excluded. Propensity score analysis was used to adjust for baseline differences. RESULTS: A total of 642 patients were enrolled. After propensity score analysis, 164 patients were classified into the DMR group, and 82 patients were classified into the AFMR group. All matched patients in both groups had atrial fibrillation. In DMR and AFMR, the 5-year freedom from readmission for heart failure and cardiac death was 96.3% in the DMR group versus 88.6% in the AFMR group (P = .045) and freedom from readmission for cardiac death in the 2 groups was 100% and 90.0%, respectively (P = .002). The recurrence rate of significant mitral regurgitation (MR) after mitral valve repair was not significantly different between the 2 groups (P = .699, log-rank test), and the 5-year freedom from MR recurrence (moderate or greater) was 89.8% and 93.0%, respectively. After the maze procedure, significantly more patients in the AFMR group than the DMR group were in junctional rhythm (49.1% vs 3.3%; P < .001) and required permanent pacemaker insertion during the follow-up period (11.4% vs 1.5% after 5 years; P = .041, log-rank test). CONCLUSIONS: AFMR was associated with acceptable outcomes of mitral valve surgery, and mitral valve repair is a good treatment option. However, significantly more patients were in junctional rhythm after the maze procedure, needing more permanent pacemaker insertion.