Effect of vitrification on protein O-GlcNAcylation in mouse metaphase II oocytes.

In brief: Oocyte vitrification leads to DNA hypomethylation, which results in defect in early embryo development. This study reveals that oocyte vitrification impairs the DNA methylation pattern by influencing protein O-GlcNAcylation. Abstract: Oocyte vitrification leads to decreased DNA methylation levels, which impairs the quality and the developmental potential of oocytes. However, the underlying molecular mechanism still need to be further revealed. In this study, mouse metaphase II (M II) oocytes were frozen by vitrification technology, while fresh oocytes were used as the control group. The effect of oocyte vitrification on protein O-GlcNAcylation and its impact on the developmental potential of oocytes were elucidated. We found that the protein O-GlcNAcylation levels were significantly increased in vitrified oocytes. Increase of protein O-GlcNAcylation levels in control oocytes by PUGNAc (an O-GlcNAcase inhibitor) decreases blastocyst rate after parthenogenetic activation (20.82% in PUGNAc-treated group; 53.82% in control group, P < 0.05). We also discovered that DNA methylation was disrupted in two-cell embryos derived from vitrified oocytes, resulting in decreased 5mC and increased 5hmC, showing similar phenotypes to that derived from PUGNAc-treated oocytes. In vitrified and PUGNAc-treated oocytes, O-GlcNAcylated TET3 was significantly increased. Notably, by inhibiting protein O-GlcNAcylation in vitrified oocytes using OSMI1 (an O-GlcNAc transferase inhibitor) we restored the DNA methylation in two-cell embryos and ameliorated the developmental defects in early embryo. Thus, elevated protein O-GlcNAcylation in vitrified oocytes is an essential contributor to their declining embryonic developmental potential. Modulation of protein O-GlcNAcylation improves the developmental potential of vitrified oocytes.

Food Environment Index is Inversely Associated with Gastric Cancer Incidence in the United States.

The first epidemiologic study was conducted to prospectively examine the association between Food Environment Index (FEI) and gastric cancer (GC) risk in the US. Surveillance, Epidemiology, and End Results provided information on GC incident cases diagnosed between 2000 and 2015 from 16 population-based cancer registries across the US. The county-level food environment was assessed using the FEI, an indicator of access to healthy foods (0 is worst, 10 is best). Poisson regression was used to calculate incidence rate ratios (IRRs) and 95% confidence intervals (CIs) for the association between FEI and GC risk adjusting for individual-level and county-level covariates. Higher levels of FEI were associated with a statistically significant reduced risk for GC (n = 87,288 cases; adjusted IRR for every score increase = 0.50, 95% CI 0.35, 0.70; P < 0.001; adjusted IRR for the medium vs. low category = 0.87, 95% CI 0.81, 0.94; and adjusted IRR for the high vs. low category = 0.89, 95% CI 0.82, 0.95). These results suggest that a healthy food environment, as measured by FEI, may be a protective factor for GC in the US. To reduce the GC incidence, further strategies to improve food environment at the county level are warranted.

Clinical Evaluation of Femoral Head Fractures: Which Classification Systems Have the Best Universality, Reliability, and Reproducibility?

BACKGROUND: Femoral head fractures are rare but potentially disabling injuries, and classifying them accurately and consistently can help surgeons make good choices about their treatment. However, there is no consensus as to which classification of these fractures is the most advantageous; parameters that might inform this choice include universality (the proportion of fractures that can be classified), as well as, of course, interobserver and intraobserver reproducibility. QUESTIONS/PURPOSES: (1) Which classification achieves the best universality (defined as the proportion of fractures that can be classified)? (2) Which classification delivers the highest intraobserver and interobserver reproducibility in the clinical CT assessment of femoral head fractures? (3) Based on the answers to those two questions, which classifications are the most applicable for clinical practice and research? METHODS: Between January 2011 and January 2023, 254 patients with femoral head fractures who had CT scans (CT is routine at our institution for patients who have experienced severe hip trauma) were potentially eligible for inclusion in this study, which was performed at a large Level I trauma center in China. Of those, 9% (23 patients) were excluded because of poor-quality CT images, unclosed physes, pathologic fractures, or acetabular dysplasia, leaving 91% (231 patients with 231 hips) for analysis here. Among those, 19% (45) were female. At the time of injury, the mean age was 40 ± 17 years. All fractures were independently classified by four observers according to the Pipkin, Brumback, AO/Orthopaedic Trauma Association (OTA), Chiron, and New classifications. Each observer repeated his classifications again 1 month later to allow us to ascertain intraobserver reliability. To evaluate the universality of classifications, we characterized the percentage of hips that could be classified using the definitions offered in each classification. The kappa (κ) value was calculated to determine interrater and intrarater agreement. We then compared the classifications based on the combination of universality and interobserver and intraobserver reproducibility to determine which classifications might be recommended for clinical and research use. RESULTS: The universalities of the classifications were 99% (228 of 231, Pipkin), 43% (99 of 231, Brumback), 94% (216 of 231, AO/OTA), 99% (228 of 231, Chiron), and 100% (231 of 231, New). The interrater agreement was judged as almost perfect (κ 0.81 [95% CI 0.78 to 0.84], Pipkin), moderate (κ 0.51 [95% CI 0.44 to 0.59], Brumback), fair (κ 0.28 [95% CI 0.18 to 0.38], AO/OTA), substantial (κ 0.79 [95% CI 0.76 to 0.82], Chiron), and substantial (κ 0.63 [95% CI 0.58 to 0.68], New). In addition, the intrarater agreement was judged as almost perfect (κ 0.89 [95% CI 0.83 to 0.96]), substantial (κ 0.72 [95% CI 0.69 to 0.75]), moderate (κ 0.51 [95% CI 0.43 to 0.58]), almost perfect (κ 0.87 [95% CI 0.82 to 0.91]), and substantial (κ 0.78 [95% CI 0.59 to 0.97]), respectively. Based on these findings, we determined that the Pipkin and Chiron classifications offer near-complete universality and sufficient interobserver and intraobserver reproducibility to recommend them for clinical and research use, but the other classifications (Brumback, AO/OTA, and New) do not. CONCLUSION: Based on our findings, clinicians and clinician-scientists can use either the Pipkin or Chiron classification systems to classify femoral head fractures based on CT images, with equal confidence. It seems unlikely that any new classifications will substantially outperform these, and the other available systems either lacked sufficient universality or reproducibility to recommend them for general use. LEVEL OF EVIDENCE: Level III, diagnostic study.

Alkaline Dilution Alters Sperm Motility in Dairy Goat by Affecting sAC/cAMP/PKA Pathway Activity.

In dairy goat farming, increasing the female kid rate is beneficial to milk production and is, therefore, economically beneficial to farms. Our previous study demonstrated that alkaline incubation enriched the concentration of X-chromosome-bearing sperm; however, the mechanism by which pH affects the motility of X-chromosome-bearing sperm remains unclear. In this study, we explored this mechanism by incubating dairy goat sperm in alkaline dilutions, examining the pattern of changes in sperm internal pH and Ca2+ concentrations and investigating the role of the sAC/cAMP/PKA pathway in influencing sperm motility. The results showed that adding a calcium channel inhibitor during incubation resulted in a concentration-dependent decrease in the proportion of spermatozoa with forward motility, and the sperm sAC protein activity was positively correlated with the calcium ion concentration (r = 0.9972). The total motility activity, proportion of forward motility, and proportion of X-chromosome-bearing sperm decreased (p < 0.05) when cAMP/PKA protease activity was inhibited. Meanwhile, the enrichment of X-chromosome-bearing sperm by pH did not affect the sperm capacitation state. These results indicate that alkaline dilution incubation reduces Ca2+ entry into X-sperm and the motility was slowed down through the sAC/cAMP/PKA signaling pathway, providing a theoretical foundation for further optimization of the sex control method.

Deep Learning Assisted Diagnosis of Musculoskeletal Tumors Based on Contrast-Enhanced Magnetic Resonance Imaging.

BACKGROUND: Misdiagnosis of malignant musculoskeletal tumors may lead to the delay of intervention, resulting in amputation or death. PURPOSE: To improve the diagnostic efficacy of musculoskeletal tumors by developing deep learning (DL) models based on contrast-enhanced magnetic resonance imaging and to quantify the improvement in diagnostic performance obtained by using these models. STUDY TYPE: Retrospective. POPULATION: Three hundreds and four musculoskeletal tumors, including 212 malignant and 92 benign lesions, were randomized into the training (n = 180), validation (n = 62) and testing cohort (n = 62). FIELD STRENGTH/SEQUENCE: A 3 T/T1 -weighted (T1 -w), T2 -weighted (T2 -w), diffusion-weighted imaging (DWI), and contrast-enhanced T1-weighted (CET1 -w) images. ASSESSMENT: Three DL models based, respectively, on the sagittal, coronal, and axial MR images were constructed to predict the malignancy of tumors. Blinded to the prediction results, a group of specialists made independent initial diagnoses for each patient by reading all image sequences. One month after the initial diagnoses, the same group of doctors made another round of diagnoses knowing the malignancy of each tumor predicted by the three models. The reference standard was the pathological diagnosis of malignancy. STATISTICAL TESTS: Sensitivity, specificity, and accuracy (all with 95% confidential intervals [CI]) corresponding to each diagnostic test were computed. Chi-square tests were used to assess the differences in those parameters with and without DL models. A P value < 0.05 was considered statistically significant. RESULTS: The developed models significantly improved the diagnostic sensitivities of two oncologists by 0.15 (95% CI: 0.06-0.24) and 0.36 (95% CI: 0.24-0.28), one radiologist by 0.12 (95% CI: 0.04-0.20), and three of the four orthopedists, respectively, by 0.12 (95% CI: 0.04-0.20), 0.29 (95% CI: 0.18-0.40), and 0.23 (95% CI: 0.13-0.33), without impairing any of their diagnostic specificities (all P > 0.128). DATA CONCLUSION: The DL models developed can significantly improve the performance of doctors with different training and experience in diagnosing musculoskeletal tumors. EVIDENCE LEVEL: 3 TECHNICAL EFFICACY: Stage 2.

Reoperation rate and implants' position variation features of displaced femoral neck fractures with sliding compression or length-stable fixation in young and middle-aged population.

INTRODUCTION: Sliding compression fixation and length-stable fixation are two basic internal fixation concepts in the treatment of displaced femoral neck fractures. In this study, we aimed to compare the reoperation rates for different methodologies of internal fixation for femoral neck fractures in young and middle-aged population. MATERIALS AND METHODS: This a retrospective study. A total of 215 patients with displaced femoral neck fractures treated with cannulated screw fixation were enrolled and divided into the sliding compression and length-stable groups according to the fixation pattern. The occurrence of and reason for revision surgery within one year were recorded. Forty-five patients with complete CT data (including CT scanning on the first postoperative day and at the last follow up) were selected from the total sample. A newly established computerized image processing method was used to evaluate variations in the spatial location of screws. RESULTS: The reoperation rate was significantly higher in the length-stable group (23.8%) than in the sliding compression group (7.3%). The rate of revision surgery due to nonunion was also higher in the length-stable group (11.4%) than in the sliding compression group (1.8%). However, no significant difference was observed in terms of joint penetration or soft tissue irritation. The sliding compression group (6.58 ± 3.18 mm) showed higher femoral neck shortening than length-stable group (4.16 ± 3.65 mm). When analyzing the spatial variations, a significantly greater screw withdrawal distance was observed in the sliding compression group than in the length-stable group, but with a smaller rotation angle. CONCLUSION: Length-stable internal fixation of displaced femoral neck fractures may lead to an increased reoperation rate in young and middle-aged population. TRIAL REGISTRATION: Name of the registry: Chinese Clinical Trial Registry. TRIAL REGISTRATION NUMBER: ChiCTR2000032327. Trial registration date: 2020-4-26.

Cortical allograft strut augmented with platelet-rich plasma for the treatment of long bone non-union in lower limb- a pilot study.

BACKGROUND: The autogenous iliac bone graft is the first choice of surgical treatment for long bone non-union. However, many factors limit the use of autogenous bone, such as insufficient bone harvest and complications in the donor site. This study aimed to pilot-test the effectiveness of the cortical allograft strut augmented with Platelet-rich plasma (PRP) on long bone non-union in the lower limb. METHOD: This study was a one-armed pilot trial, with thirteen men and four women patients scheduled for surgery. Revision surgery for managing long bone non-union included debridement, internal fixation of the cortical allograft strut, and adding PRP in the fracture site. After surgery, outcome measurements of healing rate, healing time, the incidence of revision, and complications, were assessed at least one-year follow-up. RESULTS: Fourteen of seventeen participants completed all follow-ups. The mean age of 14 patients was 35.9 years (range, 18-56 years), and the mean BMI was 22.44 ± 1.53 kg/m2. All nonunions united after the operation. The mean healing time was 4.6 ± 0.7 months. There was no revision or complication. CONCLUSION: Cortical allograft strut augmented with PRP led to healing long bone non-union in the lower limb. More clinical research is required before widespread use.

Fracture line distribution in femoral head fractures: a complement to Pipkin, Brumback, and AO/OTA classifications.

PURPOSE: This study aimed to explore the fracture line distribution and validate fracture classifications of Femoral head fractures (FHFs). MATERIALS AND METHODS: A total of 209 FHFs were reviewed retrospectively. Subjects were classified by associated injuries and commonly used fracture classifications (Pipkin, Brumback, and AO/OTA), and the universality degree of classifications was evaluated. The fracture line directions were determined in the coronal and axial CT planes. 3D mapping analysis of fracture lines was performed separately by each group. 3D maps were employed to analyze the discrimination degree of inter-subtype classifications and create a new classification. The subjects were subsequently reclassified. Correlations between classifications were analyzed to determine the matching degree of the three classifications. RESULTS: The universality degrees were 98.6% (pipkin), 44.5% (Brumback), and 94.3% (AO/OTA). The cases of (100%) Brumback and (98.5%) AO/OTA can be classified by Pipkin. The mean angles of fracture lines to the coronal and axial axis of primary compressive trabeculae were 20.25° and 54.56°. The discrimination degrees of inter-subtype of classifications were 0 (Pipkin), 60% (Brumback), and 33% (AO/OTA). A new classification with three regions and five types was created on 3D maps. Pipkin and AO/OTA matched one region, while Brumback matched two regions. CONCLUSIONS: There were three distributed fracture regions in FHFs that mismatched Pipkin, Brumback, and AO/OTA classifications. The new classification, based on morphometric features of FHFs, could compensate for the shortcomings of commonly used classifications, improving their applicability in treating FHFs.

Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes.

BACKGROUND: Oocyte vitrification has been widely used in the treatment of infertility and fertility preservation. However, vitrification-induced mitochondrial damage adversely affects oocyte development. Several studies have reported that mitochondrial calcium uptake protein 1 (MICU1) regulates the uptake of mitochondrial calcium by the mitochondrial calcium uniporter (MCU) and subsequently controls aerobic metabolism and oxidative stress in mitochondria, but research considering oocytes remains unreported. We evaluated whether the addition of MICU1 modulators enhances mitochondrial activity, pyruvate metabolism, and developmental competence after warming of MII oocytes. METHODS: Retrieved MII oocytes of mice were classified as vitrified or control groups. After thawing, oocytes of vitrified group were cultured with or without DS16570511 (MICU1 inhibitor) and MCU-i4 (MICU1 activator) for 2 h. RESULTS: Mitochondrial Ca2+ concentration, pyruvate dephosphorylation level, and MICU1 expression of MII oocytes were significantly increased after vitrification. These phenomena were further exacerbated by the addition of MCU-i4 and reversed by the addition of DS16570511 after warming. However, the mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) in vitrified-warmed MII oocytes drop significantly after vitrification, which was improved after MCU-i4 treatment and decreased significantly after DS16570511 treatment. The vitrification process was able to elicit a development competence reduction. After parthenogenetic activation, incubation of the thawed oocytes with MCU-i4 did not alter the cleavage and blastocyst rates. Moreover, incubation of the thawed oocytes with DS16570511 reduced the cleavage and blastocyst rates. CONCLUSIONS: MICU1-mediated increasing mitochondrial calcium uptake after vitrification of the MII oocytes promoted the pyruvate oxidation, and this process may maintain oocyte development competence by compensating for the consumption of ATP under stress state.

Effect of inoculation with newly isolated thermotolerant ammonia-oxidizing bacteria on nitrogen conversion and microbial community during cattle manure composting.

Nitrogen loss during composting is closely related to NH4+-N conversion, and ammonia-oxidizing bacteria (AOB) are important microorganisms that promote NH4+-N conversion. Since the biological activity of conventional AOB agents used for compost inoculation declines rapidly during the thermophilic phase of composting, new compound inoculants should be developed that are active during that phase. In the current study, the effects of inoculating cattle manure compost with newly isolated AOB (5%, v/w) [thermotolerant AOB X-2 strain (T-AOB-2), mesophilic AOB X-4 strain (M-AOB-4), and AOB X-2 combined with AOB X-4 (MT-AOB-2-4)] on the conversion of nitrogen, compost maturity, and the resident microbial community were studied. During 35 days of composting, compared with the control, AOB inoculation reduced NH3 emissions by 29.98-46.94%, accelerated the conversion of NH4+-N to NO2--N, increased seed germination values by 13.00-25.90%, and increased the abundance of the microbial community at the thermophilic phase (16.38-68.81%). Network analysis revealed that Bacillaceae play a crucial role in the composting process, with the correlation coefficients: 0.83 (p < 0.05) with NH3, 0.64 (p < 0.05) with NH4+-N, and 0.81 (p < 0.05) with NO2--N. In addition, inoculation with MT-AOB-2-4 notably increased the total nitrogen content of compost, prolonged the sanitation stage, and promoted compost maturity. Hence, MT-AOB-2-4 may be used to increase the microbial community abundance and improve the efficiency of cattle manure composting.

Fruit and Vegetable Intake is Inversely Associated with Cancer Risk in Mexican-Americans.

Objective: There is inconsistent evidence and limited data in the Hispanic population concerning fruit and vegetable intake and cancer risk. This study explored the effect of fruit and vegetable intake on cancer risk in Mexican-Americans. Methods: Participants in this cross-sectional study were drawn from the Cameron County Hispanic Cohort. Consumption of fruits and vegetables were assessed using a validated questionnaire. Cancer was self-reported by the participants based on being told by a health care provider that they had cancer. Results: Among 2,381 participants with available dietary data, 82 reported a diagnosis of cancer. Participants who met recommendations of five or more servings of fruit and vegetable per day had a significantly 86% lower risk for reported cancer compared with those who did not meet recommendations, after adjusting for other covariates. Every portion increment of total fruit and vegetable intake was significantly associated with the reduced cancer risk by 11% with the adjustment of other covariates. Conclusions: Fruit and vegetable intake was inversely associated with cancer risk in Mexican-Americans. Improving the consumption of fruit and vegetable might be an effective area for further research as part of a strategy for cancer prevention and control among Mexican-Americans independent of other factors.

A New Look at Genetic and Environmental Architecture on Lipids Using Non-Normal Structural Equation Modeling in Male Twins: The NHLBI Twin Study.

This study examined genetic and environmental influences on the lipid concentrations of 1028 male twins using the novel univariate non-normal structural equation modeling (nnSEM) ADCE and ACE models. In the best fitting nnSEM ADCE model that was also better than the nnSEM ACE model, additive genetic factors (A) explained 4%, dominant genetic factors (D) explained 17%, and common (C) and unique (E) environmental factors explained 47% and 33% of the total variance of high-density lipoprotein cholesterol (HDL-C). The percentage of variation explained for other lipids was 0% (A), 30% (D), 34% (C) and 37% (E) for low-density lipoprotein cholesterol (LDL-C); 30, 0, 31 and 39% for total cholesterol; and 0, 31, 12 and 57% for triglycerides. It was concluded that additive and dominant genetic factors simultaneously affected HDL-C concentrations but not other lipids. Common and unique environmental factors influenced concentrations of all lipids.

Circulating MicroRNAs and Life Expectancy Among Identical Twins.

Human life expectancy is influenced not only by longevity assurance mechanisms and disease susceptibility loci but also by the environment, gene-environment interactions, and chance. MicroRNAs (miRNAs) are a class of small noncoding RNAs closely related to genes. Circulating miRNAs have been shown as promising noninvasive biomarkers in the development of many pathophysiological conditions. However, the concentration of miRNA in the circulation may also be affected by environmental factors. We used a next-generation sequencing platform to assess the association of circulating miRNA with life expectancy, for which deaths are due to all causes independent of genes. In addition, we showed that miRNAs are present in 41-year archived plasma samples, which may be useful for both life expectancy and all-cause mortality risk assessment. Plasma miRNAs from nine identical male twins were profiled using next-generation sequencing. The average absolute difference in the minimum life expectancy was 9.68 years. Intraclass correlation coefficients were above 0.4 for 50% of miRNAs. Comparing deceased twins with their alive co-twin brothers, the concentrations were increased for 34 but decreased for 30 miRNAs. Identical twins discordant in life expectancy were dissimilar in the majority of miRNAs, suggesting that environmental factors are pivotal in miRNAs related to life expectancy.

Increased pSmad2 expression and cytoplasmic predominant presence of TGF-ßRII in breast cancer tissue are associated with poor prognosis: results from the Shanghai Breast Cancer Study.

Perturbations of transforming growth factor-beta (TGF-ß) signaling are pivotal to tumorigenesis and tumor progression through their effects on cell proliferation and cell invasion. This study aims to evaluate the association of TGF-ßRII and pSmad2 protein expressions in breast tissue with clinicopathological factors and prognosis of breast cancer. Expression of the TGF-ßRII and pSmad2 proteins was assessed in breast tissue of 1,045 breast cancer cases in the Shanghai Breast Cancer Study using a double immunofluorescence staining method, which was validated with standard single immunostains. TGF-ßRII expression intensity was positively associated with younger age at diagnosis (P = 0.03), pre-menopausal status (P = 0.03), and lower TNM stage (P = 0.04). Cytoplasmic predominant expression pattern of TGF-ßRII was associated with older age at diagnosis (P = 0.04) and invasive histological type (P = 0.03). Increased pSmad2 expression was associated with higher breast cancer grade (P < 0.01). Higher pSmad2 expression [HR (95 % CI):1.48 (1.07-2.04), P = 0.02] and cytoplasmic predominant TGF-ßRII expression [HR (95 % CI): 1.80 (1.08-3.00), P = 0.02] were significantly associated with reduced cancer-free survival. Our data suggest that TGF-ßRII and pSmad2 expressions are associated with certain clinical and pathologic features of breast cancer. A cytoplasmic predominant TGF-ßRII expression pattern and a higher pSmad2 expression were associated with decreased breast cancer survival. Our study provides additional evidence to support the important role of TGF-ß signaling in breast cancer prognosis.

Genetic and environmental influences on the prospective correlation between systemic inflammation and coronary heart disease death in male twins.

OBJECTIVE: Because of lack of evidence, we aimed to examine to what degree low-grade systemic inflammation and coronary heart disease (CHD) death shared common genetic and environmental substrates. APPROACH AND RESULTS: From the 41-year prospective National Heart, Lung, and Blood Institute Twin Study, we included 950 middle-aged male twins at baseline (1969-1973). Low-grade systemic inflammation was measured with plasma levels of interleukin-6 (IL-6) and C-reactive protein. Univariate and bivariate structural equation models were used, adjusted for a risk score for CHD death. The score-adjusted heritability was 19% for IL-6, 27% for C-reactive protein, and 22% for CHD death. The positive phenotypic correlation of IL-6 with CHD death (radjusted=0.27; 95% confidence interval [CI], 0.08-0.43) was driven by additive genetic factors (contribution [relative contribution], 0.30 [111%]) but attenuated by unique environment (-0.03 [-11%]). The genetic correlation between IL-6 and CHD death was 0.74 (95% CI, 0.21-1.00), whereas the unique environmental correlation was -0.05 (95% CI, -0.35 to 0.25). The proportion of genetic variance for CHD death shared with that for IL-6 was 74%. The phenotypic correlation of C-reactive protein with CHD death (radjusted=0.10; 95% CI, -0.02 to 0.22) was explained by additive genetic factors (0.20 [149%]) but was attenuated by the unique environment (-0.09 [-49%]). The genetic correlation of C-reactive protein with CHD death was 0.63 (95% CI, -0.07 to 1.00), whereas the unique environmental correlation was -0.07 (95% CI, -0.29 to 0.17). CONCLUSIONS: Low-grade systemic inflammation, measured by IL-6, and long-term CHD death share moderate genetic substrates that augment both traits.

Heritability of usual alcohol intoxication and hangover in male twins: the NAS-NRC Twin Registry.

BACKGROUND: Alcohol consumption is influenced by heritable factors. The genetic influence on usual high-density drinking, including alcohol intoxication and hangover, is unknown. We aim to estimate the heritability of usual high-density drinking. METHODS: A total of 13,511 male twins in this cross-sectional study were included from the National Academy of Sciences-National Research Council (NAS-NRC) Twin Registry. Data on the frequency of alcohol intoxication and alcohol hangover over the past year, that is, usual high-density drinking (phenotypes), were collected through a self-administered questionnaire when twins were middle-aged in 1972. Structural equation modeling was used to estimate the variance components of phenotypes. RESULTS: The mean of the frequency of usual high-density drinking in the entire twin population was 0.16 times per month for intoxication and 0.18 times per month for hangover. The heritability of usual alcohol intoxication was 50.7% (95% confidence interval [CI] 46.2 to 55.0) before and 49.9% (95% CI 45.3 to 54.2) after the body mass index (BMI) adjustment. The heritability of usual hangover was 55.4% (95% CI 51.2 to 58.6) before and 54.8% (95% CI 50.6 to 58.8) after adjustment for BMI. Unshared environmental factors between co-twins explained the remaining variance in alcohol intoxication and in hangover. CONCLUSIONS: Both genetic and unshared environmental factors have important influences on usual alcohol intoxication and hangover. These findings are important in understanding the occurrence of and developing interventions for usual high-density drinking.

Neural Manifold Constraint for Spike Prediction Models Under Behavioral Reinforcement.

Spike prediction models effectively predict downstream spike trains from upstream neural activity for neural prostheses. Such prostheses could potentially restore damaged neural communication pathways using predicted patterns to guide electrical stimulations on downstream. Since the ground truth of downstream neural activity is unavailable for subjects with the damage, reinforcement learning (RL) with behavior-level rewards becomes necessary for model training. However, existing models do not involve any constraint on the generated firing patterns and neglect the correlations among neural activities. Thus, the model outputs can greatly deviate from the natural range of neural activities, causing concerns for clinical usage. This study proposes the neural manifold constraint to solve this problem, shaping RL-generated spike trains in the feature space. The constraint terms describe the first and second order statistics of the neural manifold estimated from neural recordings during subjects' freely moving period. Then, the models can be optimized within the neural manifold by behavioral reinforcement. We test the method to predict primary motor cortex (M1) spikes from medial prefrontal (mPFC) spikes when rats perform the two-lever discrimination task. Results show that the neural activity generated by constrained models resembles the real M1 recordings. Compared with models without constraints, our approach achieves similar behavioral success rates, but reduces the mean squared error of neural firing by 61%. The constraints also increase the model's robustness across data segments and induce realistic neural correlations. Our method provides a promising tool to restore transregional communication with high behavioral performance and more realistic microscopic patterns.

Goat milk exosomal microRNAs alleviate LPS-induced intestinal inflammation in mice.

Intestinal inflammation is a common digestive system disease. Milk-derived exosomes can participate in intercellular communication and transport a variety of bioactive components, and the microRNAs (miRNAs) they carry play important roles in a variety of biological processes in the body. At present, the preventive effect and mechanism of action of goat milk exosomes and their derived miRNAs on intestinal inflammation are still unclear. In this study, the protective effect of goat milk exosomes on LPS-induced intestinal inflammation was investigated using mouse intestinal inflammation model and IEC-6 cell inflammation model. Small RNA sequencing was used to analyze the miRNA expression profile of goat milk exosomes. In this study, C-Exo and M-Exo alleviated intestinal inflammation by reducing the LPS-induced release of proinflammatory cytokines, inhibiting the increase in the NLRP3 protein and the activation of the TLR4/NFκB signaling pathway. C-Exo has a more significant inhibitory effect on them, and better therapeutic efficacy than M-Exo. Notably, the target genes of miRNAs in C-Exo and M-Exo were significantly enriched in immune-related pathways. Furthermore, their derived miR-26a-5p and miR-30a-5p were found to ameliorate the IEC-6 inflammatory response. These findings suggest that miRNAs in goat milk exosomes have the potential to attenuate LPS-induced intestinal inflammation.

Correction: Childhood cancer survival in the highly vulnerable population of South Texas: A cohort study.

[This corrects the article DOI: 10.1371/journal.pone.0278354.].

Goat Milk Exosomes Ameliorate Ulcerative Colitis in Mice through Modulation of the Intestinal Barrier, Gut Microbiota, and Metabolites.

Goat milk is rich in a variety of nutrients that are important for intestinal health and disease prevention. However, the role of exosomes in goat milk remains to be elucidated. This study investigated for the first time the therapeutic efficacy and molecular underlying mechanisms of mature milk exosomes (M-exo) and goat colostrum exosomes (C-exo) on dextran sodium sulfate-induced ulcerative colitis (UC) in mice. The findings demonstrate that M-exo and C-exo significantly improved physiological indices, suppressed the secretion of proinflammatory cytokines, and diminished oxidative stress and apoptosis in UC mice. Moreover, C-exo and M-exo restored the intestinal barrier function, remodeled the gut microbiota, and improved metabolite composition in the feces of colitis mice. In conclusion, goat milk exosomes ameliorate UC in mice, which provides a basis for the development of functional food applications for the prevention and treatment of inflammatory bowel disease.