AMPK Suppression Due to Obesity Drives Oocyte mtDNA Heteroplasmy via ATF5-POLG Axis.

Due to the exclusive maternal transmission, oocyte mitochondrial dysfunction reduces fertility rates, affects embryonic development, and programs offspring to metabolic diseases. However, mitochondrial DNA (mtDNA) are vulnerable to mutations during oocyte maturation, leading to mitochondrial nucleotide variations (mtSNVs) within a single oocyte, referring to mtDNA heteroplasmy. Obesity (OB) accounts for more than 40% of women at the reproductive age in the USA, but little is known about impacts of OB on mtSNVs in mature oocytes. It is found that OB reduces mtDNA content and increases mtSNVs in mature oocytes, which impairs mitochondrial energetic functions and oocyte quality. In mature oocytes, OB suppresses AMPK activity, aligned with an increased binding affinity of the ATF5-POLG protein complex to mutated mtDNA D-loop and protein-coding regions. Similarly, AMPK knockout increases the binding affinity of ATF5-POLG proteins to mutated mtDNA, leading to the replication of heteroplasmic mtDNA and impairing oocyte quality. Consistently, AMPK activation blocks the detrimental impacts of OB by preventing ATF5-POLG protein recruitment, improving oocyte maturation and mitochondrial energetics. Overall, the data uncover key features of AMPK activation in suppressing mtSNVs, and improving mitochondrial biogenesis and oocyte maturation in obese females.

MBNL1-AS1 Promotes Hypoxia-Induced Myocardial Infarction via the miR-132-3p/RAB14/CAMTA1 Axis.

Background: Mounting evidence have indicated that long noncoding RNA (lncRNA) muscleblind like splicing regulator 1 antisense RNA 1 (MBNL1-AS1) play a crucial regulatory role in cardiovascular disease, myocardial infarction (MI) included. In this research, we sought to probe into the biological function and potential mechanism of MBNL1-AS1 in MI. Methods: Cardiomyocytes were treated under hypoxic conditions for 0-12 h. Functional assays including CCK-8 and flow cytometry were performed to assess hypoxia-stimulated cardiomyocyte viability and apoptosis, respectively. Moreover, bioinformatics analysis and mechanical assays were conducted to reveal the competitive endogenous RNA (ceRNA) mechanism of MBNL1-AS1. Results: The upregulation of MBNL1-AS1 was found in hypoxia-stimulated cardiomyocytes. Functionally, the downregulation of MBNL1-AS1 dramatically promoted hypoxia-induced cardiomyocyte viability and inhibited apoptosis. Mechanistically, miR-132-3p bound to MBNL1-AS1 in hypoxia-induced cardiomyocytes, and miR-132-3p directly targeted RAB14, member RAS oncogene family (RAB14) and calmodulin binding transcription activator 1 (CAMTA1). Furthermore, MBNL1-AS1 upregulates the expression of RAB14 and CAMTA1 in hypoxia-stimulated cardiomyocytes via targeting miR-132-3p. Conclusions: The current study revealed the critical role of the MBNL1-AS1/miR-132-3p/RAB14/CAMTA1 axis in MI, indicating MBNL1-AS1 as an innovative therapeutic target for MI.

Vitamin a supplementation during pregnancy in shaping child growth outcomes: A meta-analysis.

Abnormal fetal growth increases risks of childhood health complications. Vitamin A supplementation (VAS) is highly accessible, but literature inconsistency regarding effects of maternal VAS on fetal and childhood growth outcomes exists, deterring pregnant women from VAS during pregnancy. This meta-analysis aimed to analyze effects of vitamin A only or vitamin A + co-intervention during pregnancy in healthy mothers (MH) or with complications (MC, night blindness and HIV positive) on perinatal growth outcomes, also assess VAS dose impacts. The Cochrane Library, PubMed, ScienceDirect, Scopus, Embase and Web of Science databases were searched from inception to July 15, 2021. We covered subgroup analyses, including VAS in MH or MC within randomized controlled trial (RCT) or observational studies (OS). Fifty-five studies were included in this meta-analysis (426,098 pregnancies). Vitamin A decreased risk of preterm birth by 9% in MH-RCT (P < 0.001), by 62% in MH-OS (P = 0.029), by 10% in MC-RCT (P = 0.089); decreased LBW by 24% in MC-RCT (P = 0.032); increased neonatal weight in MC-RCT (SMD 0.96; P = 0.051). Besides, vitamin A + co-intervention decreased risks of preterm by 18% in MH-OS (P = 0.021); LBW by 25% in MH-OS (P < 0.001); by 32% in MC-RCT (P = 0.006); decreased neonatal defects by 33% in MH-OS (P = 0.064); decreased anemia by 25% in MH-OS (P = 0.0003); increased neonatal weight in MH-OS (SMD 0.51; P = 0.014); and increased neonatal length in MH-OS (SMD 1.83; P = 0.013). Meta-regression of VAS dose with individual outcomes was not significant, and no side effects were observed for VAS doses up to 4000 mcg (RAE/d). Regardless of maternal health conditions, VAS during pregnancy can safely and effectively improve fetal development and neonatal health even in mothers without VAD.

Impact of autoantibodies against the M2-muscarinic acetylcholine receptor on clinical outcomes in peripartum cardiomyopathy patients with standard treatment.

OBJECTIVES: To evaluate the impact of autoantibodies against the M2-muscarinic receptor (anti-M2-R) on the clinical outcomes of patients receiving the standard treatment for peripartum cardiomyopathy (PPCM). METHODS: A total of 107 PPCM patients who received standard heart failure (HF) treatment between January 1998 and June 2020 were enrolled in this study. According to anti-M2-R reactivity, they were classified into negative (n = 59) and positive (n = 48) groups, denoted as the anti-M2-R (-) and anti-M2-R (+) groups. Echocardiography, 6-min walk distance, serum digoxin concentration (SDC), and routine laboratory tests were performed regularly for 2 years. The all-cause mortality, cardiovascular mortality, and rehospitalisation rate for HF were compared between the two groups. RESULTS: A total of 103 patients were included in the final data analysis, with 46 in the anti-M2-R (+) group and 57 in the anti-M2-R (-) group. Heart rate was lower in the anti-M2-R (+) group than in the anti-M2-R (-) group at the baseline (102.7 ± 6.1 bpm vs. 96.0 ± 6.4 bpm, p < 0.001). The initial SDC was higher in the anti-M2-R (+) group than in the anti-M2-R (-) group with the same dosage of digoxin (1.25 ± 0.45 vs. 0.78 ± 0.24 ng/mL, p < 0.001). The dosages of metoprolol and digoxin were higher in the anti-M2-R (-) patients than in the anti-M2-R (+) patients (38.8 ± 4.6 mg b.i.d. vs. 27.8 ± 5.3 mg b.i.d., p < 0.0001, respectively, for metoprolol; 0.12 ± 0.02 mg/day vs. 0.08 ± 0.04 mg/day, p < 0.0001, respectively, for digoxin). Furthermore, there was a greater improvement in cardiac function in the anti-M2-R (-) patients than in the anti-M2-R (+) patients. Multivariate analysis identified negativity for anti-M2-R as the independent predictor for the improvement of cardiac function. Rehospitalisation for HF was lower in the anti-M2-R (-) group, but all-cause mortality and cardiovascular mortality were the same. CONCLUSIONS: There were no differences in all-cause mortality or cardiovascular mortality between the two groups. Rehospitalisation rate for HF decreased in the anti-M2-R (-) group. This difference may be related to the regulation of the autonomic nervous system by anti-M2-R.

Effects of Maternal Exercise During Pregnancy on Perinatal Growth and Childhood Obesity Outcomes: A Meta-analysis and Meta-regression.

BACKGROUND: Perinatal growth abnormalities program susceptibility to childhood obesity, which is further exaggerated by maternal overweight and obesity (MO) during pregnancy. Exercise is highly accessible, but reports about the benefits of maternal exercise on fetal growth and childhood obesity outcomes are inconsistent, reducing the incentives for pregnant women to participate in exercise to improve children's perinatal growth. OBJECTIVE: This systematic review and meta-analysis aims to establish evidence-based efficacy of exercise in mothers with normal weight (MNW) and MO during pregnancy in reducing the risks of perinatal growth abnormalities and childhood obesity. In addition, the impacts of exercise volume are also assessed. METHODS: The PubMed, ScienceDirect, Web of Science, and Cochrane Library databases were searched from inception to February 15, 2020. We included randomized controlled trials with exercise-only intervention or exercise with other confounders in pregnant MNW (body mass index, BMI 18.5-24.9 kg/m2) and MO (BMI ≥ 25 kg/m2), which were further subgrouped in the meta-analysis. Primary outcomes included birth weight, preterm birth, small for gestational age (SGA), large for gestational age (LGA), infant and childhood weight, and childhood obesity. A linear meta-regression analysis was also used to explore the effects of exercise volume on outcomes. RESULTS: 99 studies were included in the meta-analysis (n = 596,876), and individual study quality ranged from fair to good according to the Newcastle-Ottawa scale assessment. Exercise only interventions in MNW reduced preterm birth by 15% (26 studies, n = 76,132; odds ratio [OR] 0.85; 95% CI 0.72, 1.01; I2 = 83.3%), SGA by 17% (33 studies, n = 92,351; OR 0.83; 95% CI 0.71, 0.98; I2 = 74.5%) and LGA by 17% (29 studies, n = 84,310; OR 0.83; 95% CI 0.74, 0.95; I2 = 60.4%). Exercise only interventions in MO reduced preterm birth by 33% (2 studies, n = 3,050; OR 0.67; 95% CI 0.70, 0.96; I2 = 0%), SGA by 27% (8 studies, n = 3,909; OR 0.73; 95% CI 0.50, 1.05; I2 = 40.4%) and LGA by 55% (9 studies, n = 81,581; OR 0.45; 95% CI 0.18, 1.11; I2 = 98.3%). Exercise only interventions in MNW reduced childhood obesity by 53% (3 studies, n = 6,920; OR 0.47; 95% CI 0.36, 0.63; I2 = 77.0%). However, no significant effect was observed in outcomes from exercise confounders in either MNW or MO. In the meta-regression, the volume of exercise-only intervention in MNW was negatively associated with birth weight, greatly driven by volumes more than 810 metabolic equivalents (MET)-min per week. Other outcomes were not associated with exercise volume. CONCLUSIONS: This systematic review and meta-analysis suggests that exercise during pregnancy in both MNW and MO safely and effectively reduce the risks of preterm birth, SGA, and LGA. Furthermore, MNW exercise also reduces the risk of childhood obesity. Overall, regardless of prepregnancy BMI, maternal exercise during pregnancy provides an excellent opportunity to mitigate the high prevalence of adverse birth outcomes and childhood obesity.

Methane yields during anaerobic co-digestion of animal manure with other feedstocks: A meta-analysis.

Anaerobic co-digestion of animal manure with other feedstocks (aka co-digestion) is increasingly being used to enhance methane yield and organic waste management. The benefits accruing from co-digestions compared to mono-digestions, however, vary greatly in the literature. The goal of this research was to use meta-analysis to critically compare methane yields between mono- and co-digestions and identify relevant factors (co-substrate type, substrate dose, carbon to nitrogen (C/N) ratio, volatile solids (VS), substrate pH, organic loading rate (OLR), and hydraulic retention time (HRT)) contributing to methane yield. Published studies (n = 64 representing 384 case-studies) with sufficient detail on methane yield were identified for the meta-analysis. Analysis indicated that co-digestion of animal manure with other feedstocks significantly increased methane yield (249 L kg-1[VS]), compared with anaerobic mono-digestion of animal manure (171 L kg-1[VS]). Similar methane yields increases (47-57 L kg-1[VS]) were obtained from co-digestions in batch reactors of swine (238-287 L kg-1[VS]), poultry (213-260 L kg-1[VS]), and cattle manure (147-204 L kg-1[VS]). In continuous digesters of cattle manure (175-299 L kg-1[VS]) co-digestion had the greatest methane yield improvement of 124 L kg-1[VS], swine manure (212-322 L kg-1[VS]) co-digestion ranked second with 110 L kg-1[VS], and poultry manure ranked third with 70 L kg-1[VS]. Improved methane yield were obtained at optimum C/N ratio ranging from 26 to 34. The respective optimum OLR for co-digestion of swine, poultry, and cattle manure were 1.2, 1.4 and 3.4 kg VS m-3 d-1, while the recommended HRT was 30-40 d. Taken together, anaerobic co-digestion of animal manure with other feedstock significantly improved anaerobic digestion. Factors contributing to methane yields included: substrate-type and dose, VS, C/N, OLR, and HRT.

Effect of stearic or oleic acid on milk performance and energy partitioning when fed in diets with low and high rumen-active unsaturated fatty acids in early lactation.

This experiment was conducted to determine the effects of stearic acid (SA; C18:0) or rumen-protected oleic acid (OA; C18:1 cis-9) on milk performance and energy partitioning of early lactation cows when supplemented in diets with low and high level of rumen unsaturated fatty acids (RUFA). In low RUFA experiment (LRUFA), FA supplement rich in either SA or calcium salts OA was added to a basal diet with a low concentration of RUFA (0.75% vs. 1.4%, LRUFA-SA vs. LRUFA-OA). In high RUFA experiment (HRUFA), 2% soybean oil was added to the diet fed in the LRUFA experiment. In each experiment, 30 multiparous cows were blocked by parity and predicted transmitting ability for milk yield and were randomly fed 1 of 2 treatment diets from 2 to 13 wk postpartum. In the LRUFA experiment, LRUFA-SA had 2.4 kg/d more dry matter intake (DMI) (P < 0.01), 3.8 kg/d more energy-corrected milk (P < 0.01), and 0.3% units more milk fat percentage (P < 0.01) and 0.2 kg/d more milk fat yield (P < 0.01). Dietary treatments did not affect body weight, energy balance, and energy intake partitioning into milk, maintenance, and body tissues (P > 0.1). In the HRUFA experiment, HRUFA-SA had 1.4 kg/d more DMI (P = 0.03) but similar milk and milk components yields (P > 0.1). HRUFA-SA had a tendency to gain more body weight (P = 0.07) and had more positive energy balance (P = 0.01) and decreased gross feed efficiency (milk yield/DMI) (P = 0.01). Consistently, HRUFA-SA increased intake energy partitioning into body tissues (P = 0.02) and decreased energy partitioning into milk (P = 0.01). In summary, SA supplementation had more DMI relative to OA, but the effects on milk and milk fat production were different and affected by the level of RUFA in the basal diet. In application, SA supplementation was more effective to improve milk production when included in the basal diet with the low RUFA.

Association of Autoantibodies against M2-Muscarinic Acetylcholine Receptor with Atrial Fibrosis in Atrial Fibrillation Patients.

OBJECTIVES: To investigate the association of serum autoantibodies against M2-muscarinic acetylcholine receptor (anti-M2-R) with atrial fibrosis in long-standing persistent atrial fibrillation (AF) patients. METHODS: Twenty-four long-standing persistent AF patients, scheduled to undergo hybrid ablation surgery, were enrolled in the study. Twenty-six patients with sinus rhythm, scheduled to undergo coronary artery bypass grafting surgery, were enrolled into the non-AF group. We detected serum anti-M2-R levels. Left atrial appendages were subjected to histological and molecular biological assays. Patients in the AF group received follow-up for two years. RESULTS: The AF group showed significantly higher serum anti-M2-R levels compared to the non-AF group (496.2 ± 232.5 vs. 86.3 ± 25.7 pmol/L, p < 0.001). The AF group exhibited severe fibrosis in the left atrial appendages, as indicated by increased collagen volume fraction (45.2 ± 4.7% vs. 27.6 ± 8.3%, p < 0.001), and higher levels of collagen I (0.52 ± 0.04 vs. 0.24 ± 0.06, p < 0.001) and collagen III (0.51 ± 0.07 vs. 0.36 ± 0.09, p < 0.001). TGF-ß1 and CTGF were also upregulated in the AF group. A positive correlation between serum anti-M2-R levels and fibrosis of the left atrial appendage and fibrogenic indexes was observed. CONCLUSIONS: Serum anti-M2-R levels are higher in AF patients and are associated with the severity of atrial fibrosis. In addition, serum anti-M2-R levels are positively correlated to TGF-ß1 and CTGF expression in the left atrial appendage.

Association of autoantibodies against the M2-muscarinic receptor with long-term outcomes in peripartum cardiomyopathy patients: A 5-year prospective study.

BACKGROUND: Peripartum cardiomyopathy (PPCM) is characterized by heart failure. Our previous study found that autoantibodies against the M2-muscarinic receptor (anti-M2-R) are increased in PPCM patients. We aimed to evaluate the association of anti-M2-R on prognosis of PPCM patients with standard treatment. METHODS: Synthetic peptides corresponding to the M2 receptor served as the target antigens in an enzyme-linked immunosorbent assay experiment. They were used to screen the sera of 80 PPCM patients, who were separated into anti-M2-R-negative and positive groups according to their anti-M2-R reactivity. Clinical assessment and echocardiography examination were performed at baseline and after 5 years with a standard treatment regimen. The endpoint events were compared after 5 years of follow-up. RESULTS: There were 76 PPCM patients who completed the final data analysis, including 36 in the anti-M2-R (+) group and 40 in the anti-M2-R (-) group. Both groups showed improvement in the left ventricular end-diastolic and end-systolic dimensions and the ejection fraction with standard treatment regimens for 5 years (all p<0.001). Patients in the anti-M2-R (-) group had greater tolerance and were more rapidly titrated to metoprolol, and they had better improvement in cardiac function than patients in the anti-M2-R (+) group (p<0.05). Patients in the anti-M2-R (-) group had a marked decrease in re-hospitalization (p<0.05), but not in all-cause mortality or cardiovascular mortality. Being positive for anti-M2-R increased the risk of PPCM (OR=4.7, 95% CI 1.8-12.2, p=0.002). CONCLUSIONS: PPCM patients, especially anti-M2-R (-) patients, have a relatively better prognosis than other patients. We posit that the presence of anti-M2-R may be involved in the pathogenesis of PPCM.

The Effect of Autoantibody against M2-Muscarinic Acetylcholine Receptor in Heart Failure Patients on Digoxin Treatment.

BACKGROUND: Autoantibody against M2-muscarinic acetylcholine receptor (anti-M2AChR) has a biological effect similar to a vagus agonist. Digoxin has a function of vagus nervous system stimulation. We hypothesized that anti-M2AChR is highly correlated with digoxin in patients with chronic heart failure (CHF). METHODS: Synthetic M2AChR peptides served as the target antigen in an ELISA were used to screen the sera of 80 CHF patients, who were separated into a negative (-) or positive (+) anti-M2AChR group according to their anti-M2AChR reactivity. Echocardiography and serum digoxin concentration (SDC) were performed at baseline and after 1 year of digoxin in combination with the standard treatment regime. The end-point events were compared over 1 year of follow-up. RESULTS: Seventy-two CHF patients completed the final data analysis, including 32 (+)anti-M2AChR and 40 (-)anti-M2AChR patients. The resting heart rate of the positive group was higher than that of the negative group at baseline (p < 0.05; 89.0 ± 1.6 vs. 83.8 ± 1.1 bpm). Both groups showed improvement in the left ventricular end-diastolic and end-systolic dimensions and ejection fraction with digoxin in combination with the standard treatment regime for 1 year (all p < 0.01). However, the 32 patients with (-)anti-M2AChR had greater improvements than the 40 patients with (+)anti-M2AChR, and this was accompanied by a marked decrease of rehospitalization (all p < 0.01) but not of cardiovascular mortality after 1 year. The SDC of patients with (-)anti-M2AChR was significantly lower than that of patients with (+)anti-M2AChR (p < 0.05; 0.63 ± 0.05 vs.1.16 ± 0.06 ng/mL) and had a positive correlation with anti-M2AChR (r = 0.81, p < 0.001). CONCLUSION: These results suggested that anti-M2AChR could be a useful biomarker of vagus nerve overactivation and is associated with a poor response to digoxin treatment in CHF patients.

Nutrient contributions and biogas potential of co-digestion of feedstocks and dairy manure.

This study focused on collection of data on nutrient flow and biogas yield at a commercial anaerobic digester managed with dairy manure from a 1000 cow dairy and co-digestion of additional feedstocks. Feedstocks included: blood, fish, paper pulp, out of date beverages and grease trap waste. Mass flow of inputs and outputs, nutrient concentration of inputs and outputs, and biogas yield were obtained. It was determined that manure was the primary source of nutrients to the anaerobic digester when co-digested with feedstocks. The percentage of contribution from manure to the total nutrient inputs for total nitrogen, ammonia-nitrogen, phosphorus and total solids was 46.3%, 67.7%, 32.8% and 23.4%, respectively. On average, manure contributed the greatest amount of total nitrogen and ammonia-nitrogen. Grease trap waste contributed the greatest amount of phosphorus and total solids at approximately 50%. Results demonstrated that a reliable estimate of nutrient inflow could be obtained from the product of the nutrient analyses of a single daily composite of influent subsamples times the total daily flow estimated with an in-line flow meter. This approach to estimate total daily nutrient inflow would be more cost effective than testing and summing the contribution of individual feedstocks. Data collected after liquid-solid separation confirmed that the majority (>75%) of nutrients remain with the liquid effluent portion of the manure stream. It was demonstrated that the ash concentration in solids before and after composting could be used to estimate the mass balance of total solids during the compost process. This data confirms that biogas or methane yield could be accurately measured from the ratio of % volatile solids to % total solids.

Short communication: Temporal effect of feeding potassium carbonate sesquihydrate on milk fat in lactating dairy cows fed a fat-depressing diet.

A lactation study with 10 multiparous dairy cows in early lactation, with an average of 64 days in milk (standard deviation=37), were used to evaluate how quickly milk fat concentration would change when potassium carbonate sesquihydrate was abruptly added to the diet. The experiment had 3 periods. In period 1 (d 0 to 7) all cows were fed the same basal (control) diet with 1.8% soy oil, dry basis; in period 2 (d 8 to 28) 5 cows received the control diet, whereas the other 5 cows received the control diet plus 0.59% of added K with K carbonate sesquihydrate; and in period 3 (d 29 to 42) all 10 cows received the control diet. The control diet was formulated for a dietary cation-anion difference (DCAD), calculated as Na + K - Cl - S, of 37.7mEq/100g of dry matter (DM), 1.74% of DM as K, and 5.7% long-chain fatty acids (DM%), which included 1.8% of DM as soybean oil. Period 1 was used as a covariate. In period 2, d 8 to 28, 5 cows remained on the control diet whereas 5 cows were fed with the control diet plus K carbonate sesquihydrate (DCAD+ diet; DCAD of 54.3mEq/100g DM and 2.33% of DM as K). After feeding the DCAD+ diet, we noted a difference in milk fat concentration from 3.9 to 4.3% within 72h. Over the 21d of period 2, the DCAD+ diet resulted in significantly greater milk fat percentage from 4.0 to 4.3%, lactose from 4.74 to 4.82%, and fat efficiency in the form of fat in milk divided by fat in DMI from 1.27 to 1.49, without affecting dry matter intake (DMI), milk protein concentration, solids-not fat concentration, 3.5% fat-corrected milk, and protein efficiency in the form of protein in milk divided by protein in DMI. In period 3 (d 29-42), all cows were again fed the control diet, resulting in a tendency for greater milk fat concentration, significantly greater lactose concentration, and fat efficiency in the form of fat in milk divided by fat in DMI for the cows having received the DCAD+ diet during period 2. In conclusion, the abrupt addition of K carbonate sesquihydrate resulted in a greater milk fat concentration and tended to maintain the greater concentration after cessation of K carbonate sesquihydrate feeding.

Adipositas cordis sudden death: a series of 79 patients.

BACKGROUND: The principal aim of this study was to investigate the clinical, epidemiological and pathologic features for a series of 79 cases of adipositas cordis sudden death. METHODS: We analyzed clinical and autopsy pathological features of 79 patients (43 females and 36 males) with adipositas cordis who died suddenly between 1975 and 2010. Data were extracted from China National Knowledge Infrastructure and Wan Fang Database. RESULTS: The average age of the 79 cases was 36.6 ± 1.4 years old ranging from 13 to 68, and 82.3% of them were between 20 to 50 years old. Sudden death was the first symptom in 62 (78.5%) of the cases, only 17 (21.5%) had a history of chest distress or dyspnea. More than 4/5 (87.3%) of the cases had no any past medical history. At autopsy, the subjects' heart weight was mild or moderately increased, and a large amount of fatty tissues but not fibrous or fibro fatty was accumulated underneath the epicardium and infiltrated toward the right ventricle walls, and even infiltrated to all layers of the cardiac walls. Regional epidemiological data showed that about 80% of cases were living north and only 20% were living south of the Yangzi River, but not any familial heredity. CONCLUSION: Adipositas cordis sudden death is a very severe disease, it occurs mostly in youth and middle-aged and sudden death is often the first symptom. There is a significant regional difference, but not any genetic correlation. The pathogenesis of adipositas cordis sudden death should to be further explored.

Downregulation of the miR-30 family microRNAs contributes to endoplasmic reticulum stress in cardiac muscle and vascular smooth muscle cells.

BACKGROUND: Endoplasmic reticulum (ER) stress is a common subcellular response to stresses and central to ER stress is increased expression of glucose-regulated protein 78 (GRP78). However, the mechanisms for GRP78 upregulation remained poorly understood. Our study goal was to shed light on this issue. METHODS: H2O2 was used to create cellular models of ER stress in neonatal rat ventricular cells (NRVCs) and rat aorta vascular smooth muscle cells (RAVSMCs). Molecular Biology techniques were used to quantify protein and mRNA levels. Luciferase reporter gene assay was employed to investigate miRNA targeting. MTT assay and ELISA were used to detect cell death. RESULTS: MiRNAs belonging to the miR-30 family including miR-30a, b, c, d and e were all downregulated in ER stress induced by H2O2 in cardiovascular cells NRVCs and RAVSMCs, along with the upregulation of GRP78, cleaved ATF6, CHOP, and cleaved caspase-12. GRP78 was confirmed to be a target gene for miR-30. Artificial knockdown of miR-30 by antimiR-30 triggered the phenotypic ER stress with significant GRP78/ATF6/CHOP/caspase-12 upregulations and cell death, while miR-30 replacement mitigated ER stress. Knockdown of CHOP by siRNA regulated all members of the miR-30 family whereas sequestration of C/EBP transcription factor by its decoy downregulated miR-30 miRNAs. CONCLUSIONS: Collectively, downregulation of the miR-30 family miRNAs contributes to the ER stress and the associated upregulation of GRP78 in the cardiovascular system. The participation of miR-30 creates a positive feedback loop in the ER stress signaling pathway. MiR-30 replacement may be a viable approach for alleviating disorders associated with ER stress.

Association of autoantibodies against the M2-muscarinic receptor with perinatal outcomes in women with severe preeclampsia.

BACKGROUND: The goal of this study was to test the hypothesis that autoantibodies against M2-muscarinic acetylcholine receptor (M2-AAB) are associated with severe preeclampsia and increased risk of adverse perinatal outcomes. METHODS: We conducted a case-control study comparing 60 women with severe preeclampsia to 60 women with normal pregnancy and 60 non-pregnant controls. A peptide, corresponding to amino acid sequences of the second extracellular loops of the M2 receptor, was synthesized as antigen to test for the presence of autoantibodies, using an enzyme-linked immunosorbent assay. The frequency and titer of M2-AAB were compared in the 3 groups. The risk of adverse perinatal outcomes among women with severe preeclampsia in the presence of M2-AAB was estimated. RESULTS: M2-AAB were positive in 31.7% (19/60) of patients with severe preeclampsia, in 10.0% (6/60) (p=0.006) of normal pregnant women and in 8.3% (5/60) (p=0.002) of non-pregnant controls. The presence of M2-AAB was associated with increased risk of adverse pregnancy complications (OR, 3.6; 95%CI, 1.0-12.6; p=0.048), fetal growth restriction (OR, 6.8; 95% CI, 2.0-23.0; p=0.002), fetal distress (OR, 6.7; 95% CI, 1.7-26.6; p=0.007), low Apgar score (OR, 5.3; 95% CI, 1.4-20.7; p=0.017), and perinatal death (OR, 4.3; 95% CI, 1.0-17.6; p=0.044) among women with severe preeclampsia. CONCLUSIONS: This study demonstrates, for the first time, an increase in M2-AAB in patients with severe preeclampsia. Women with severe preeclampsia who are M2-AAB positive are at increased risk for neonatal mortality and morbidity. We posit that M2-AAB may be involved in the pathogenesis of severe preeclampsia.

Association between the presence of autoantibodies against adrenoreceptors and severe pre-eclampsia: a pilot study.

BACKGROUND: Pre-eclampsia is the leading cause of maternal and neonatal morbidity and mortality with incompletely understood etiopathogenesis. The purpose of the current study is to determine whether there is a relationship between the presence of autoantibodies against ß1, ß2 and α1 adrenoreceptors and severe pre-eclampsia. METHODOLOGY/PRINCIPAL FINDINGS: Synthetic peptides corresponding to amino acid sequences of the second extracellular loops of ß1, ß2 and α1 adrenoreceptors were synthesized as antigens to test 34 patients with severe pre-eclampsia, 36 normal pregnancy women and 40 non-pregnant controls for the presence of autoantibodies using enzyme-linked immunosorbent assay. The respective frequencies of autoantibodies against ß1, ß2 and α1 adrenoreceptors were 50.0% (17/34), 52.9% (18/34) and 55.9% (19/34) in patients with severe pre-eclampsia, 19.4% (7/36) (p = 0.011), 19.4% (7/36) (p = 0.006) and 17.6% (6/36) (p = 0.001) in normal pregnancy women and 10% (4/40), 7.5% (3/40) and 10% (4/40) (p<0.001) in non-pregnant controls. Titers of these autoantibodies were also significantly increased in patients with severe pre-eclampsia. By logistic regression analysis, the presence of these three autoantibodies significantly increased the risk of neonatal death (odds ratio, 13.5; 95% confidence interval, 1.3-141.3; p = 0.030) and long-term neonatal hospitalization (odds ratio, 5.0; 95% confidence interval, 1.3-19.1; p = 0.018). The risk of hypertension and fetal distress were also associated with the presence of these three autoantibodies. CONCLUSIONS/SIGNIFICANCE: This novel pilot study demonstrated for the first time that the presence of autoantibodies against ß1, ß2 and α1 adrenoreceptors are increased in patients with severe pre-eclampsia. Pregnant women who are positive for the three autoantibodies are at increased risks of neonatal mortality and morbidity. We posit that these autoantibodies may be involved in the pathogenesis of severe pre-eclampsia.

Predictive value of pre-procedural autoantibodies against M2-muscarinic acetylcholine receptor for recurrence of atrial fibrillation one year after radiofrequency catheter ablation.

BACKGROUND: Increasing evidences have suggested that autoantibodies against muscarinic-2 acetylcholine receptor (anti-M2-R) may play an important role in the development of atrial fibrillation (AF). Predictive value of pre-procedural anti-M2-R for the recurrence of AF after radiofrequency catheter ablation is still unclear. METHODS: Totally 76 AF patients with preserved left ventricular systolic function were prospectively enrolled and subjected to ablation after the detection of serum anti-M2-R by enzyme linked immunosorbent assay. These patients were given follow-up examination for one year after ablation. Risk estimation for the recurrence of AF was performed using the univariate and multivariate logistic regression. RESULTS: In AF group, serum anti-M2-R was significantly higher than that in the control group in terms of frequency (40.8% versus 11.7%; p < 0.001) and titer (1:116 versus 1:29; p < 0.001). Compared with paroxysmal AF patients, persistent AF patients had higher frequency (57.6% versus 27.9%; p = 0.009) and titer (1:132 versus 1:94; p = 0.012) for autoantibodies. During one-year follow-up examination after ablation, the recurrence of AF was observed in 25 (32.9%) patients. Multivariate analysis showed that pre-procedural serum anti-M2-R was an independent predictor for the recurrence of AF at the time point of 12 months after ablation (odds ratio: 4.701; 95% confidence interval: 1.590-13.894; p = 0.005). CONCLUSIONS: In AF patients, the frequency and titer of serum anti-M2-R were significantly higher than those in the control group with sinus rhythm. Pre-procedural serum anti-M2-R was an independent predictor for the recurrence of AF one year after radiofrequency catheter ablation.