Essential role of glycoprotein Ibα in platelet activation.

Glycoprotein (GP) Ib, the ligand-binding subunit of platelet GPIb-IX complex, interacts with von Willebrand factor (VWF) exposed at the injured vessel wall, initiating platelet adhesion, activation, hemostasis, and thrombus formation. The cytoplasmic tail of GPIb interacts with 14-3-3, regulate ng the VWF-GPIb-elicited signal transduction and VWF binding function of GPIb. However, we unexpectedly found that the GPIb-14-3-3 association, beyond VWF-dependent function, is essential for general platelet activation. We found that the GPIb cytoplasmic tail peptide MPC, a potential GPIb inhibitor, by itself induced platelet aggregation, integrin αIIbß3 activation, granule secretion, and phosphatidylserine (PS) exposure. Conversely, the deletion of the cytoplasmic tail of GPIb in mouse platelets (10aa-/-) decreased platelet aggregation, integrin IIb3 activation, granule secretion, and PS exposure induced by various physiological agonists. Phosphoproteome-based kinase activity profiling revealed significantly upregulated protein kinase C (PKC) activity in MPC-treated platelets. MPC-induced platelet activation was abolished by the pan-PKC inhibitor and PKC deletion. Decreased PKC activity was observed in both resting and agonist-stimulated 10aa-/- platelets. GPIb regulates PKC activity by sequestering 14-3-3 from PKC. In vivo, the deletion of the GPIb cytoplasmic tail impaired mouse hemostasis and thrombus formation and protected against platelet-dependent pulmonary thromboembolism. Therefore, our findings demonstrate an essential role for the GPIb cytoplasmic tail in regulating platelet general activation and thrombus formation beyond the VWF-GPIb axis.

Correlational patterns of neuronal activation and epigenetic marks in the basolateral amygdala and piriform cortex following olfactory threat conditioning and extinction in rats.

Introduction: Cumulative evidence suggests that sensory cortices interact with the basolateral amygdala (BLA) defense circuitry to mediate threat conditioning, memory retrieval, and extinction learning. The olfactory piriform cortex (PC) has been posited as a critical site for olfactory associative memory. Recently, we have shown that N-methyl-D-aspartate receptor (NMDAR)-dependent plasticity in the PC critically underpins olfactory threat extinction. Aging-associated impairment of olfactory threat extinction is related to the hypofunction of NMDARs in the PC. Methods: In this study, we investigated activation of neuronal cFos and epigenetic marks in the BLA and PC using immunohistochemistry, following olfactory threat conditioning and extinction learning in rats. Results: We found highly correlated cFos activation between the posterior PC (pPC) and BLA. cFos was correlated with the degree of behavioral freezing in the pPC in both adult and aged rats, and in the BLA only in adult rats. Markers of DNA methylation 5 mC and histone acetylation H3K9/K14ac, H3K27ac, and H4ac exhibited distinct training-, region-, and age-dependent patterns of activation. Strong correlations of epigenetic marks between the BLA and pPC in adult rats were found to be a general feature. Conversely, aged rats only exhibited correlations of H3 acetylations between the two structures. Histone acetylation varied as a function of aging, revealed by a reduction of H3K9/K14ac and an increase of H4ac in aged brains at basal condition and following threat conditioning. Discussion: These findings underscore the coordinated role of PC and BLA in olfactory associative memory storage and extinction, with implications for understanding aging related cognitive decline.

Microbial methanogenesis in aerobic water: A key driver of surface methane enrichment in a deep reservoir.

Significant amounts of the greenhouse gas methane (CH4) are released into the atmosphere worldwide via freshwater sources. The surface methane maximum (SMM), where methane is supersaturated in surface water, has been observed in aquatic systems and contributes significantly to emissions. However, little is known about the temporal and spatial variability of SMM or the mechanisms underlying its development in artificial reservoirs. Here, the community composition of methanogens as major methane producers in the water column and the mcrA gene was investigated, and the cause of surface methane supersaturation was analyzed. In accordance with the findings, elevated methane concentration of SMM in the transition zone, with an annually methane emission flux 2.47 times higher than the reservoir average on a large and deep reservoir. In the transition zone, methanogens with mcrA gene abundances ranging from 0.5 × 103-1.45 × 104 copies/L were found. Methanobacterium, Methanoseata and Methanosarcina were the three dominate methanogens, using both acetic acid and H2/CO2 pathways. In summary, this study contributes to our comprehension of CH4 fluxes and their role in the atmospheric methane budget. Moreover, it offers biological proof of methane generation, which could aid in understanding the role of microbial methanogenesis in aerobic water.

Enantioselective and Regiodivergent Synthesis of Propargyl- and Allenylsilanes through Catalytic Propargylic C-H Deprotonation.

We report a highly enantioselective intermolecular C-H bond silylation catalyzed by a phosphoramidite-ligated iridium catalyst. Under reagent-controlled protocols, propargylsilanes resulting from C(sp3)-H functionalization, as well the regioisomeric and synthetically versatile allenylsilanes, could be obtained with excellent levels of enantioselectivity and good to excellent control of propargyl/allenyl selectivity. In the case of unsymmetrical dialkyl acetylenes, good to excellent selectivity for functionalization at the less-hindered site was also observed. A variety of electrophilic silyl sources (R3SiOTf and R3SiNTf2), either commercial or in situ-generated, were used as the silylation reagents, and a broad range of simple and functionalized alkynes, including aryl alkyl acetylenes, dialkyl acetylenes, 1,3-enynes, and drug derivatives were successfully employed as substrates. Detailed mechanistic experiments and DFT calculations suggest that an η3-propargyl/allenyl Ir intermediate is generated upon π-complexation-assisted deprotonation and undergoes outer-sphere attack by the electrophilic silylating reagent to give propargylic silanes, with the latter step identified as the enantiodetermining step.

Pressure-induced phase transition of Lu2Ti2O7and Lu1.5Ce0.5Ti2O7+xpyrochlores.

This study utilizes both experimental and computational approaches to investigate the performance of Lu2Ti2O7(LTO) and Lu1.5Ce0.5Ti2O7+x(LCTO) pyrochlores under high pressure. The structural changes of LTO and LCTO pyrochlores were characterized usingin-situsynchrotron x-ray diffraction (SXRD) andin-situRaman spectroscopy at pressures up to 44.6 GPa. The kinks inP-aandP-Vcurves at around 5 GPa are mainly attributed to the interaction between the pressure medium and the isostructural changes. The onset pressures for transitioning from the cubic pyrochlore phase (Fd-3 m) to the monoclinic phase (P21) are observed at 32.5 GPa and 38.1 GPa, respectively. It is important to note that at the highest measured pressures, the phase transition remains incomplete. This partial transition is likely the result of oriented disorder among cations and anions under high pressure. In addition, introducing Ce as a dopant significantly enhances structural stability. This can be explained by the larger ionic radius of Ce, which hinders the disordering process.

Novel missense mutation c.797T>C (p.Met266Thr) gives rise to the rare B(A) phenotype in a Chinese family.

BACKGROUND AND OBJECTIVES: B(A) phenotype is usually formed by nucleotide mutations in the ABO*B.01 allele, with their products exhibiting glycosyltransferases (GTs) A and B overlapping functionality. We herein report a B(A) allele found in a Chinese family. MATERIALS AND METHODS: The entire ABO genes of the probands, including flanking regulatory regions, were sequenced through PacBio third-generation long-read single-molecule real-time sequencing. 3D molecular models of the wild-type and mutant GTB were generated using the DynaMut web server. The effect of the mutation on the enzyme function was predicted by PROVEAN and PolyPhen2. The predictions of stability changes were performed using DynaMut and SNPeffect. RESULTS: Based on serological and sequencing features, we concluded the two probands as possible cases of the B(A) phenotype. Crystallization analysis showed that Thr266 substitution does not disrupt the hydrogen bonds. However, some changes in interatomic contacts, such as loss of ionic interactions and hydrophobic contacts, and addition of weak hydrogen bonds, may have affected protein stability to some extent. This mutation was predicted to have a benign effect on enzyme function and slightly reduce protein stability. CONCLUSION: The probands had the same novel B(A) allele with a c.797T>C (p.Met266Thr) mutation on the ABO*B.01 backbone.

"Three-in-One" Nanozyme Composite for Augmented Cascade Catalytic Tumor Therapy.

Cascade catalytic reaction exhibits simple procedure and high efficiency, such as that from the orderly assembly of different enzymes in biological systems. Mimicking of the natural cascade procedure becomes critical, but the orderly assembly of different enzymes is still challenging. Herein, single Au-Pt nanozyme is reported with "three-in-one" functions to initiate cascade conversions for O2 supply as mimic catalase, H2 O2 production with its glucose oxidase-like property, and • OH generation as mimic peroxidase for chemodynamic therapy (CDT). Thus, the complex assembly and cross-talk among the different enzymes are avoided. To this end, metastable Cu2 O NPs, as scaffolds, are used to anchor ultrasmall Au-Pt nanozyme, while metal-organic framework (MOF) is used to encapsulate the nanozyme for tumor microenvironment response and shielding protein adsorption. Pluronic F127 is then modified on the surface to improve hydrophilicity and biocompatibility of the composite. The endogenous acidity and glutathione in tumor degrade MOF to expose nanozyme for cascade catalytic CDT. The high photothermal conversion ability also enhances the CDT, while Cu2+ ions consume GSH to further improve CDT efficiency as augmented cascade catalytic tumor therapy. Thus, a new paradigm is provided with drug-free single nanozyme for improving tumor therapeutic efficacy and minimizing side effects.

Advantages of diapause in Anastatus japonicus Ashmead mass production on eggs of the Chinese oak silkworm, Antheraea pernyi.

BACKGROUND: Anastatus japonicus Ashmead, a biological control agent utilized in China to control fruit bugs and forest caterpillars, is efficiently raised on large Chinese oak silkworm (Antheraea pernyi) eggs. Here, we investigated the biological parameters of non-diapaused and diapaused Anastatus japonicus after long-term storage within eggs of the host, Antheraea pernyi, under laboratory conditions. RESULTS: Diapaused mature larvae of Anastatus japonicus were more cold-tolerant than non-diapaused mature larvae, as reflected by a lower supercoiling point, lower freezing point, and higher survival rate at cold temperatures. Diapause induction enhanced the lifespan, fecundity and oviposition period of Anastatus japonicus than non-diapaused Anastatus japonicus when refrigerated for 6 months. However, after 12 months of refrigeration, the fecundity and oviposition period of Anastatus japonicus were significantly reduced with and without diapause. No difference in the progeny sex ratio of Anastatus japonicus was observed between diapause-induction treatment and those of non-diapaused. With the extension of refrigeration period from 6 months to 12 months, the lifespan, fecundity and oviposition period of Anastatus japonicus which were treated with diapause induction showed a sharp decrease. No significantly difference in the lifespan, fecundity and oviposition period of Anastatus japonicus was observed between diapause-induction treatment and those of non-diapaused when refrigerated for 12 months. CONCLUSION: These results suggest that the induction of diapause is an applicable technique to achieve mass production of Anastatus japonicus in long-term storage using eggs of the factitious host Antheraea pernyi, without compromising the quality of the parasitoid. The refrigeration period of diapaused Anastatus japonicus should not exceed 6 months. © 2023 Society of Chemical Industry.

The role of PKC in X-ray-induced megakaryocyte apoptosis and thrombocytopenia.

Thrombocytopenia is a critical complication after radiation therapy and exposure. Dysfunction of megakaryocyte development and platelet production are key pathophysiological stages in ionizing radiation (IR)-induced thrombocytopenia. Protein kinase C (PKC) plays an important role in regulating megakaryocyte development and platelet production. However, it remains unclear how PKC regulates IR-induced megakaryocyte apoptosis. In this study, we found that pretreatment of PKC pan-inhibitor Go6983 delayed IR-induced megakaryocyte apoptosis, and inhibited IR-induced mitochondrial membrane potential and ROS production in CMK cells. Moreover, suppressing PKC activation inhibited cleaved caspase3 expression and reduced p38 phosphorylation levels, and IR-induced PKC activation might be regulated by p53. In vivo experiments confirmed that Go6983 promoted platelet count recovery after 21 days of 3 Gy total body irradiation. Furthermore, Go6983 reduced megakaryocyte apoptosis, increased the number of megakaryocyte and polyploid formation in bone marrow, and improved the survival rate of 6 Gy total body irradiation. In conclusion, our results provided a potential therapeutic target for IR-induced thrombocytopenia.

Stability and bifurcation analysis of a two-patch model with the Allee effect and dispersal.

In the current manuscript, a two-patch model with the Allee effect and nonlinear dispersal is presented. We study both the ordinary differential equation (ODE) case and the partial differential equation (PDE) case here. In the ODE model, the stability of the equilibrium points and the existence of saddle-node bifurcation are discussed. The phase diagram and bifurcation curve of our model are also given as a results of numerical simulation. Besides, the corresponding linear dispersal case is also presented. We show that, when the Allee effect is large, high intensity of linear dispersal is not favorable to the persistence of the species. We further show when the Allee effect is large, nonlinear diffusion is more beneficial to the survival of the population than linear diffusion. Moreover, the results of the PDE model extend our findings from discrete patches to continuous patches.

An eco-epidemic model for assessing the application of integrated pest management strategies.

Mathematical models have become indispensable tools for analyzing pest control strategies. However, in the realm of pest control studies, the consideration of a plant population being affected by a model that incorporates pests, natural enemies and disease in the pest population has been relatively limited. Therefore, this paper aims to formulate and investigate a hybrid impulsive eco-epidemic model that incorporates disease in the pest population. Initially, we examine the existence and stability of the pest-eradication periodic solution. Subsequently, to explore the impact of chemical and biological control methods, we propose an updated eco-epidemic model that incorporates varying frequencies of pesticide sprays and the release of both infected pests and natural enemies for pest control. We establish threshold values for the susceptible pest eradication periodic solution under different scenarios, illustrating the global attractiveness of this solution. Finally, we discuss the obtained results and suggest potential avenues for future research in this field.

Physical Activity During Adolescence and Early-adulthood and Ovarian Cancer Among Women with a BRCA1 or BRCA2 Mutation.

In the general population, physical activity has been associated with a lower risk of several cancers; however, the evidence for ovarian cancer is not clear. It is suggested that early-life physical activity may differentially impact risk. Whether this is true among women at high risk due to a pathogenic variant (mutation) in the BRCA1 or BRCA2 genes has not been evaluated. Thus, we performed a matched case-control study to evaluate the association between adolescent and early-adulthood physical activity and ovarian cancer. BRCA mutation carriers who completed a research questionnaire on various exposures and incident disease and with data available on physical activity were eligible for inclusion. Self-reported activity at ages 12-13, 14-17, 18-22, 23-29, and 30-34 was used to calculate the average metabolic equivalent of task (MET)-hours/week for moderate, vigorous, and total physical activity during adolescence (ages 12-17) and early-adulthood (ages 18-34). Conditional logistic regression was used to estimate the OR and 95% confidence intervals (CI) of invasive ovarian cancer associated with physical activity. This study included 215 matched pairs (mean age = 57.3). There was no association between total physical activity during adolescence (ORhigh vs. low = 0.91; 95% CI: 0.61-1.36; Ptrend = 0.85), early-adulthood (ORhigh vs. low = 0.78; 95% CI: 0.51-1.20; Ptrend = 0.38) and overall (ORhigh vs. low = 0.81; 95% CI: 0.54-1.23; Ptrend = 0.56) and ovarian cancer. Findings were similar for moderate (Ptrend ≥ 0.25) and vigorous (Ptrend ≥ 0.57) activity. These findings do not provide evidence for an association between early-life physical activity and BRCA-ovarian cancer; however, physical activity should continue to be encouraged to promote overall health. SIGNIFICANCE: In this matched case-control study, we observed no association between physical activity during adolescence or early-adulthood and subsequent risk of ovarian cancer. These findings do not provide evidence for an association between early-life physical activity and BRCA-ovarian cancer; however, being active remains important to promote overall health and well-being.

Polymorphisms in the promoter regions of RHD and RHCE genes in the Chinese Han population.

BACKGROUND AND OBJECTIVES: The Rh blood group system is the most polymorphic human blood group system. Previous studies have investigated variants in the RHD and RHCE promoter. The relevance of these variants to the Chinese Han population is further clarified in this study. MATERIALS AND METHODS: In total, 317 donors (223 Rh D-positive [D+], including 20 Del and 94 Rh D-negative [D-]) were randomly selected. The promoter regions and exon 1 of RHD and RHCE were amplified through polymerase chain reaction (PCR) whose products were directly sequenced using forward and reverse primers. RESULTS: Expected PCR products of the RHD promoter and exon 1 were amplified in 223 D+ individuals, including 20 Del individuals, and were absent in 81 of 94 D- individuals. Expected PCR products of RHCE were observed in all donors. Two single nucleotide variants (SNVs) were observed in the RHD promoter region. Moreover, 11 SNVs were observed in the promoter and exon 1 of RHCE. rs4649082, rs2375313, rs2281179, rs2072933, rs2072932, rs2072931 and rs586178 with strong linkage disequilibria were significantly different between the D+ and D- groups. [A;C] was the most common haplotype in the RHD promoter (NC_000001.11:g.[-1033A>G;-831C>T]). [G;T;T;A;T;A;C;G;A;C;G] was the most predominant haplotype in both total and D- groups. In D+ individuals, [A;C;T;G;C;G;C;G;C;C;C] was the most frequent haplotype in the RHCE promoter (NC_000001.11:g.[-1080A>G;-958C>T;-390T>C;-378G>A;-369C>T;-296G>A;-144C>G;-132G>A;-122C>A;28C>T;48C>G]). CONCLUSION: We speculate that the SNVs/haplotypes found in this article cannot significantly affect gene expression. The present study findings should help elucidate the molecular basis of the polymorphic expression of RHD and RHCE promoter regions.

A Novel Variant in the Desmoplakin Gene in One Case of the Rare Carvajal Syndrome with Dilated Cardiomyopathy: A Case Report and Literature Review.

Carvajal syndrome is a rare hereditary cardiocutaneous syndrome caused by the variants of the desmoplakin (DSP) gene. In this study, we report a patient of Carvajal syndrome with a novel homozygous missense variant of DSP gene. We diagnosed a 7-year-old female patient with Carvajal syndrome characterized by dilated cardiomyopathy, palmoplantar keratoderma, woolly hair, and dental dysplasia, who disclosed a novel homozygous missense variant c.4597C > T (p.Q1533X) in exon 6 of the DSP gene found for the first time. Both her parents were heterozygous for the identified nonsense variant c.4597C > T (p.Q1533X) in DSP gene but neither showed evidence of Carvajal syndrome, indicating that this novel variant causes the disease in an autosomal recessive manner. Genotypes of Carvajal syndrome are even broader than so far anticipated. When patients with dilated cardiomyopathy, palmoplantar keratoderma, woolly hair, and dental dysplasia are found in clinical practice, Carvajal syndrome should be highly suspected, and family gene sequencing should be actively carried out.

Analysis of postoperative pregnancy outcome in 180 women with congenital uterine malformation.

Introduction: This study aims to explore the effects of combination of laparoscopy and hysteroscopy in pregnancy outcome in women diagnosed with congenital uterine malformation (CUM). The observation criteria include pregnancy rate, misdiagnosis rate, rate of spontaneous abortion and preterm birth rate. Material and methods: A total of 180 patients with congenital uterine malformation, who were treated in our hospital from January 2015 to June 2018, were enrolled in the study. Prior to hospitalization, all the patients had neither a history of genital tract surgery nor endocrine abnormalities, chromosomal abnormalities, immune abnormalities or other factors affecting pregnancy. Furthermore, the ovarian functions were normal, and there were no factors leading to infertility in the male partners. The diagnosis was mainly based on medical history, clinical manifestations, gynecological examinations, and ultrasonography including two-dimensional and three-dimensional ultrasonography, as well as hysterosalpingogram (HSG), magnetic resonance imaging (MRI), hysteroscopy, and/or laparoscopy or surgery. Patients were diagnosed and classified according to the Buttram classification. Results: Among these 180 patients, 37 patients were diagnosed with complete septate uterus, 96 patients had sub-septate uterus, 25 patients had unicornuate uterus, 11 patients were diagnosed with bicornuate uterus, and 11 patients had didelphic uterus. The total number of preoperative pregnancies was 112, including 106 spontaneous abortions, with an abortion rate of 94.64%, and 86 total postoperative pregnancies, among which spontaneous abortions occurred 11 times, with an abortion rate of 12.79%. The difference was statistically significant (p < 0.05). Conclusions: Uterine malformation surgery can significantly improve the reproductive prognosis in patients with CUM.

Neferine mediated TGF-ß/ERK signaling to inhibit fibrosis in endometriosis.

OBJECTIVE: To investigate the mechanism of Neferine in treating endometriosis fibrosis by TGF-ß/ERK signaling pathway through a combination of network pharmacological analysis of Lotus embryos, in vivo animal experiments, and in vitro cell experiments. METHODS: The active ingredients of the drug lotus embryos, the drug targets and the targets of endometriosis were determined from the TCMSP database, the Swiss Target Prediction database and GeneCard and Online Mendelian Inheritance in Man. The String database and Cytoscape 3.6.3 software were used to construct the network of common target protein interactions between drug and disease, as well as the target network. GO and KEGG enrichment analysis of the common targets was performed. We designed endometriosis mouse models with Neferine to investigate the therapeutic effect of Neferine on the fibrosis model of endometriosis and its mechanism of action. Different methods were used to evaluate the treated endometriotic lesion tissue and the untreated ectopic lesion tissue. The 12Z cells (human endometriosis immortalized cells) were cultured in vitro and treated with Neferine to detect cell viability and the effects of invasion and metastasis. RESULTS: The results of GO function and KEGG enrichment analysis showed that the role pathways of lotus germ were TGF-ß signaling pathway, ERK1/2 signaling pathway, IL-17 signaling pathway, TNF signaling pathway, AGE-RAGE signaling pathway, and PI3K-Akt signaling pathway. Neferine which is one of the effective active ingredients of lotus germ, significantly inhibited the expression of fibronectin, collagen I, connective tissue growth factor, and smooth muscle actin by activating the TGF-ß/ERK pathway in vivo, which is required for the fibrosis process of endometriosis. Neferine also significantly inhibited the proliferation, invasion and metastasis ability of 12Z cells. CONCLUSION: Neferine inhibits the progression of endometriosis both in vitro and in vivo. Its mechanism of action may involve the regulation of the TGF-ß/ERK signaling pathway, leading to the inhibition of fibrosis in endometriosis.

Association between radiomics features of DCE-MRI and CD8+ and CD4+ TILs in advanced gastric cancer.

Objective: The aim of this investigation was to explore the correlation between the levels of tumor-infiltrating CD8+ and CD4+ T cells and the quantitative pharmacokinetic parameters of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) in patients with advanced gastric cancer. Methods: We retrospectively analyzed the data of 103 patients with histopathologically confirmed advanced gastric cancer (AGC). Three pharmacokinetic parameters, Kep, Ktrans, and Ve, and their radiomics characteristics were obtained by Omni Kinetics software. Immunohistochemical staining was used to determine CD4+ and CD8+ TILs. Statistical analysis was subsequently performed to assess the correlation between radiomics characteristics and CD4+ and CD8+ TIL density. Results: All patients included in this study were finally divided into either a CD8+ TILs low-density group (n = 51) (CD8+ TILs < 138) or a high-density group (n = 52) (CD8+ TILs ≥ 138), and a CD4+ TILs low-density group (n = 51) (CD4+ TILs < 87) or a high-density group (n = 52) (CD4+ TILs ≥ 87). ClusterShade and Skewness based on Kep and Skewness based on Ktrans both showed moderate negative correlation with CD8+ TIL levels (r = 0.630-0.349, p < 0.001), with ClusterShade based on Kep having the highest negative correlation (r = -0.630, p < 0.001). Inertia-based Kep showed a moderate positive correlation with the CD4+ TIL level (r = 0.549, p < 0.001), and the Correlation based on Kep showed a moderate negative correlation with the CD4+ TIL level, which also had the highest correlation coefficient (r = -0.616, p < 0.001). The diagnostic efficacy of the above features was assessed by ROC curves. For CD8+ TILs, ClusterShade of Kep had the highest mean area under the curve (AUC) (0.863). For CD4+ TILs, the Correlation of Kep had the highest mean AUC (0.856). Conclusion: The radiomics features of DCE-MRI are associated with the expression of tumor-infiltrating CD8+ and CD4+ T cells in AGC, which have the potential to noninvasively evaluate the expression of CD8+ and CD4+ TILs in AGC patients.