Subclass phenotypes in patients with unprovoked venous thromboembolisms using a latent class analysis.

BACKGROUND: Patients with unprovoked venous thromboembolisms (VTEs) can be sub-classified based on the different phenotypes using a latent class analysis (LCA), which might be useful for selecting individual management strategies. METHODS: In the COMMAND VTE Registry-2 database enrolling 5197 VTE patients, the current derivation cohort consisted of 1556 patients with unprovoked VTEs. We conducted clustering with an LCA, and the patients were classified into subgroups with the highest probability. We compared the clinical characteristics and outcomes among the developed subgroups. RESULTS: This LCA model proposed 3 subgroups based on 8 clinically relevant variables, and classified 592, 813, and 151 patients as Class I, II, and III, respectively. Based on the clinical features, we named Class I the younger, Class II the older with a few comorbidities, and Class III the older with many comorbidities. The cumulative 3-year anticoagulation discontinuation rate was highest in the older with many comorbidities (Class III) (39.9 %, 36.1 %, and 48.4 %, P = 0.02). There was no significant difference in the cumulative 5-year incidence of recurrent VTEs among the 3 classes (12.8 %, 11.1 %, and 4.0 % P = 0.20), whereas the cumulative 5-year incidence of major bleeding was significantly higher in the older with many comorbidities (Class III) (7.8 %, 12.7 %, and 17.8 %, P = 0.04). CONCLUSION: The current LCA revealed that patients with unprovoked VTEs could be sub-classified into further phenotypes depending on the patient characteristics. Each subclass phenotype could have different clinical outcomes risks especially a bleeding risk, which could have a potential benefit when considering the individual anticoagulation strategies. CLINICAL TRIAL REGISTRATION: URL: http://www.umin.ac.jp/ctr/index.htm COMMAND VTE Registry-2: Unique identifier, UMIN000044816 COMMAND VTE Registry: Unique identifier, UMIN000021132.

Nardilysin in vascular smooth muscle cells controls blood pressure via the regulation of calcium dynamics.

Blood pressure is a crucial physiological parameter and its abnormalities can cause a variety of health problems. We have previously reported that mice with systemic deletion of nardilysin (NRDC), an M16 family metalloprotease, exhibit hypotension. In this study, we aimed to clarify the role of NRDC in vascular smooth muscle cell (VSMC) by generating VSMC-specific Nrdc knockout (VSMC-KO) mice. Our findings reveal that VSMC-KO mice also exhibit hypotension. Aortas isolated from VSMC-KO mice exhibited a weakened contractile response to phenylephrine, accompanied by reduced phosphorylation of myosin light chain 2 and decreased rhoA expression. VSMC isolated from VSMC-KO aortas showed a reduced increase in intracellular Ca2+ concentration induced by α-stimulants. These findings suggest that NRDC in VSMC regulates vascular contraction and blood pressure by modulating Ca2+ dynamics.

Prospective study on planned biliary stent placement to treat small common bile duct stones.

Aims: Small common bile duct stones are known to occasionally clear spontaneously. This study aimed to prospectively assess the role of biliary stent placement in promoting the spontaneous clearance of small common bile duct stones. Methods and Results: We analyzed patients presenting with common bile duct stones of ≤5 mm diameter between June 2020 and May 2022. The exclusion criteria included asymptomatic patients, biliary pancreatitis, altered gastrointestinal anatomy, bile duct strictures (malignant or benign), and a history of EST. The biliary stents were inserted without stone removal. Stone clearance was assessed using endoscopic ultrasonography or endoscopic retrograde cholangiopancreatography after 3 months. Our primary endpoint was the clearance rate of common bile duct stones over 6 months, targeting a lower limit for the 95% confidence interval (CI) exceeding 25%. Of the 32 enrolled patients, 18 (56.3%; 95% CI: 37.7-73.6%) exhibited stone clearance. Early complications occurred in 11 patients (34.4%), totaling 12 incidents: acute cholecystitis in four, acute pancreatitis in three, biliary pain in three, and cholangitis in two patients. No severe complications occurred. Six (18.8%) patients experienced asymptomatic stent migration. Following stone clearance, four (12.5%) patients experienced stone recurrence, with an average duration of 256 ± 164 days. Conclusion: Biliary stenting appeared to effectively promote the clearance of small common bile duct stones in approximately half of the patients. However, the potential complications and risks of stone recurrence warrant close monitoring.This trial was registered in the Japan Registry of Clinical Trials (jRCT1042200020).

Col1α2-Cre-mediated recombination occurs in various cell types due to Cre expression in epiblasts.

The Cre-LoxP system has been commonly used for cell-specific genetic manipulation. However, many Cre strains exhibit excision activity in unexpected cell types or tissues. Therefore, it is important to identify the cell types in which recombination takes place. Fibroblasts are a cell type that is inadequately defined due to a lack of specific markers to detect the entire cell lineage. Here, we investigated the Cre recombination induced by Col1α2-iCre, one of the most common fibroblast-mesenchymal Cre driver lines, by using a double-fluorescent Cre reporter line in which GFP is expressed when recombination occurs. Our results indicated that Col1α2-iCre activity was more extensive across cell types than previously reported: Col1α2-iCre-mediated recombination was found in not only cells of mesenchymal origin but also those of other lineages, including haematopoietic cells, myocardial cells, lung and intestinal epithelial cells, and neural cells. In addition, study of embryos revealed that recombination by Col1α2-iCre was observed in the early developmental stage before gastrulation in epiblasts, which would account for the recombination across various cell types in adult mice. These results offer more insights into the activity of Col1α2-iCre and suggest that experimental results obtained using Col1α2-iCre should be carefully interpreted.

An adult female case of autoimmune pancreatitis with gastric outlet obstruction complicated by annular pancreas.

A 65-year-old woman presented with epigastric pain persisting for more than 3 months. She was diagnosed with autoimmune pancreatitis (AIP), based on high serum IgG4 levels (981 mg/dL) and diffuse pancreatic enlargement with a capsule-like rim on computed tomography (CT). Additionally, the main pancreatic duct was indistinct on magnetic resonance cholangiopancreatography. CT, esophagogastroduodenoscopy, and upper gastrointestinal radiography revealed stenosis with gastric outlet obstruction (GOO) in the second part of the duodenum. Prednisolone administration was initiated as treatment; on day 3 of treatment, the patient's symptoms improved. After 2 weeks, CT and endoscopic ultrasonography of the duodenal bulbs revealed improvement of the enlarged pancreas. The second part of the duodenum ran into the pancreatic head, and no malignant lesions were observed. Based on the above findings, we suspect that she developed AIP in the annular pancreas (AnnP), where duodenal stenosis worsened with diffuse pancreatic enlargement, resulting in GOO. She is currently under careful observation with tapering of prednisolone-without surgical treatment for AnnP. The pathogenesis of GOO caused by AIP without malignancy is rare. One case of GOO caused by AIP, wherein AIP developed in the AnnP (similar to the present case), has been reported, highlighting the novelty of our report.

Upregulated Ribosomal Pathway Impairs Follicle Development in a Polycystic Ovary Syndrome Mouse Model: Differential Gene Expression Analysis of Oocytes.

Polycystic ovary syndrome (PCOS), a common endocrine disorder, is associated with impaired oocyte development, leading to infertility. However, the pathogenesis of PCOS has not been completely elucidated. This study aimed to determine the differentially expressed genes (DEGs) and epigenetic changes in the oocytes from a PCOS mouse model to identify the etiological factors. RNA-sequencing analysis revealed that 90 DEGs were upregulated and 27 DEGs were downregulated in mice with PCOS compared with control mice. DNA methylation analysis revealed 30 hypomethylated and 10 hypermethylated regions in the PCOS group. However, the DNA methylation status did not correlate with differential gene expression. The pathway enrichment analysis revealed that five DEGs (Rps21, Rpl36, Rpl36a, Rpl37a, and Rpl22l1) were enriched in ribosome-related pathways in the oocytes of mice with PCOS, and the immunohistochemical analysis revealed significantly upregulated expression levels of Rps21 and Rpl36. These results suggest that differential gene expression in the oocytes of mice in PCOS is related to impaired folliculogenesis. These findings improve our understanding of PCOS pathogenesis.

Rupture of Non-aneurysmal Mycotic Thoracic Aortic Arteritis Caused by Campylobacter fetus.

Campylobacter fetus (C. fetus) demonstrates a preference for vascular tissue and is an infrequent etiology of mycotic aortic arteritis (MAA), mostly occurring in the abdominal aorta. MAA characteristically has a rapid progression to aneurysm formation and subsequently, to aortic rupture. We present a 73-year-old woman with non-aneurysmal mycotic thoracic aortic arteritis (MTAA) complicated with a rupture caused by C. fetus. She presented after four days of pain in the lower abdomen. Contrast-enhanced computed tomography revealed non-aneurysmal descending thoracic aorta arteritis and an abdominal aorta aneurysm, and the blood cultures were positive for C. fetus. Antibiotic therapy relieved the abdominal pain. However, eight days after the antibiotic therapy, she died because of a rupture of the non-aneurysmal MTAA. The non-aneurysmal MTAA caused by C. fetus ruptured while the infection was being treated with appropriate antibiotics, and there was no sign of arterial dilatation. An early open or endovascular repair after a short pre-operative antibiotic therapy may be required for non-aneurysmal MAA caused by C. fetus. More cases of non-aneurysmal MAA caused by C. fetus are needed to determine the clinical course and to decide the treatment strategy.

Smooth muscle protein 22α-Cre recombination in resting cardiac fibroblasts and hematopoietic precursors.

The Cre-loxP system has been widely used for cell- or organ-specific gene manipulation, but it is important to precisely understand what kind of cells the recombination takes place in. Smooth muscle 22α (SM22α)-Cre mice have been utilized to alter genes in vascular smooth muscle cells (VSMCs), activated fibroblasts or cardiomyocytes (CMs). Moreover, previous reports indicated that SM22α-Cre is expressed in adipocytes, platelets or myeloid cells. However, there have been no report of whether SM22α-Cre recombination takes place in nonCMs in hearts. Thus, we used the double-fluorescent Cre reporter mouse in which GFP is expressed when recombination occurs. Immunofluorescence analysis demonstrated that recombination occurred in resting cardiac fibroblasts (CFs) or macrophages, as well as VSMCs and CMs. Flow cytometry showed that some CFs, resident macrophages, neutrophils, T cells, and B cells were positive for GFP. These results prompted us to analyze bone marrow cells, and we observed GFP-positive hematopoietic precursor cells (HPCs). Taken together, these results indicated that SM22α-Cre-mediated recombination occurs in resting CFs and hematopoietic cell lineages, including HPCs, which is a cautionary point when using SM22α-Cre mice.

Effects of constitutively active IKKß on cardiac development.

NF-κB is a major transcription factor regulating cell survival, organ development and inflammation, but its role in cardiac development has been inadequately explored. To examine this function, we generated mice in which IKKß, an essential kinase for NF-κB activation, was constitutively activated in embryonic cardiomyocytes. For this purpose, we used smooth muscle-22α (SM22α)-Cre mice, which are frequently used for gene recombination in embryonic cardiomyocytes. Embryonic hearts of SM22αCre-CA (constitutively active) IKKßflox/flox mice revealed remarkably thin, spongy and hypoplastic myocardium. In exploring the mechanism, we found that the expression of bone morphogenetic protein 10 (BMP10) and T-box transcription factor 20 (Tbx20), major regulators of cardiac development, was significantly downregulated and upregulated, respectively, in the SM22αCre-CAIKKßflox/flox mice. We also generated NK2 homeobox 5 (Nkx2.5) Cre-CAIKKßflox/wt mice since Nkx2.5 is also expressed in embryonic cardiomyocytes and confirmed that the changes in these genes were also observed. These results implicated that the activation of NF-κB affects cardiac development.

Disruption of piRNA machinery by deletion of ASZ1/GASZ results in the expression of aberrant chimeric transcripts in gonocytes.

In the male germline, the machinery to repress retrotransposons that threaten genomic integrity via the piRNA pathway is established in gonocytes. It has been reported that disruption of the piRNA pathway leads to activation of retrotransposons and arrests spermatogenesis before it enters the second meiosis; however, its effects on gonocytes have not been fully elucidated. In this study, we analyzed the effects of Asz1 deletion, which is a crucial component of the piRNA pathway, on the gonocyte transcriptome. In Asz1-null gonocytes, MIWI2, which is responsible for introducing DNA methylation to retrotransposons in a piRNA-dependent manner, disappeared from the nuclei of fetal gonocytes. Transcriptome analysis revealed that retrotransposons targeted by the piRNA pathway and non-annotated transcript variants were upregulated in gonocytes from neonatal Asz1-/- mice. These non-annotated transcript variants were chimeras generated by joining exons transcribed from retrotransposons and canonical genes. DNA methylation analysis showed that retrotransposons that induce the expression of aberrant chimeric transcripts are not fully methylated. This was consistent with the impaired nuclear localization of MIWI2 in Asz1-null gonocytes. Furthermore, heterogeneity of DNA methylation status in retrotransposons was observed in both gonocytes and their descendants. This suggests that the piRNA system in gonocytes can potentially prevent spermatogenic cell populations bearing aberrant chimeric transcripts from propagating later in spermatogenesis. In conclusion, Asz1 is required to repress retrotransposons and retrotransposon-driven aberrant chimeric transcripts in gonocytes through the piRNA pathway.

Insights into in vivo follicle formation: a review of in vitro systems.

In vitro systems capable of reconstituting the process of mouse oogenesis are now being established to help develop further understanding of the mechanisms underlying oocyte/follicle development and differentiation. These systems could also help increase the production of useful livestock or genetically modified animals, and aid in identifying the causes of infertility in humans. Recently, we revealed, using an in vitro system for recapitulating oogenesis, that the activation of the estrogen signaling pathway induces abnormal follicle formation, that blocking estrogen-induced expression of anti-Müllerian hormone is crucial for normal follicle formation, and that the production of α-fetoprotein in fetal liver tissue is involved in normal in vivo follicle formation. In mouse fetuses, follicle formation is not carried out by factors within the ovaries but is instead orchestrated by distal endocrine factors. This review outlines findings from genetics, endocrinology, and in vitro studies regarding the factors that can affect the formation of primordial follicles in mammals.

[Comparison of two cases of type 2 autoimmune pancreatitis].

We describe two cases of type 2 autoimmune pancreatitis (AIP). A 39-year-old man presented to our hospital with complaints of epigastric and back pain. Pancreatic enzyme levels were elevated, but serum levels of immunoglobulins G and G4 (IgG and IgG4) were normal. Computed tomography (CT) showed diffuse pancreatic enlargement, and endoscopic retrograde pancreatography revealed diffuse narrowing of the pancreatic duct. Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) revealed granulocytic epithelial lesions and very few IgG4-positive cells. Colonoscopy revealed ulcerative colitis. Type 2 AIP was diagnosed, and 5-aminosalicylic acid (5-ASA) and prednisolone were administered. The clinical course has since been favorable, and the prednisolone dose is currently being reduced. A 47-year-old woman presented to our hospital with complaints of bloody stools. Colonoscopy revealed ulcerative colitis. CT depicted diffuse pancreatic enlargement with a capsule-like rim. Pancreatic enzyme levels were elevated, but serum levels of IgG and IgG4 were normal. On magnetic resonance cholangiopancreatography, the pancreatic duct could not be delineated. No pathological findings of type 2 AIP were obtained on EUS-FNA. Type 2 AIP was suspected, and 5-ASA and steroid enemas were administered. To date, recurrence has not been observed, and 5-ASA management continues. The two cases differed with regard to sex of patient, clinical course, pathological findings, and treatment.

Optimal conditions for mouse follicle culture.

Mammalian ovaries contain a large number of immature follicles. Follicular culture can contribute to the production of fertile oocytes from latent immature follicles, providing a useful tool for exploring the developmental competencies and related factors that oocytes acquire during growth. However, the potential of oocytes produced by follicular culture is limited. Herein, the optimal follicular culture conditions for the addition of polyvinylpyrrolidone to the medium and oxygen concentration were investigated. Polyvinylpyrrolidone with a high molecular weight (≥ 360,000) and a 7% oxygen concentration were found to increase the blastocyst formation rate by more than 20% compared with conventional culture conditions. Although the developmental ability of oocytes produced by follicular culture remained inferior to that of in vivo-derived oocytes, these findings may pave the way for enhanced production of fertile oocytes in vitro and for studying the process of full developmental potency acquisition by oocytes.

Correlations between the solubility and surface characteristics of milk protein concentrate powder particles.

The solubility of high-protein milk protein concentrate (MPC) may decrease significantly during storage, particularly at relatively high temperatures and humidity. The objective of this study was to seek correlations between the solubility loss of MPC during storage and various surface characteristics determined on the basis of simultaneous nanoscale topographical imaging and nanomechanical mapping of MPC particle surfaces using atomic force microscopy. A control MPC and a calcium-depleted MPC were stored at 45°C and 66% relative humidity for up to 60 d. The solubility of the control MPC was 56% at the beginning of the storage and gradually decreased to 10% at the end of the 60-d storage. The calcium-depleted MPC exhibited more rapid decreases from almost 100% at the beginning of the storage to 18% after storage for 45 d, after which we observed no significant difference in solubility between the control and calcium-depleted MPC. Averaged or root mean squared roughness values calculated using topographical images were found to have no correlation with the solubility. Deformation, Derjaguin-Muller-Toropov modulus, and adhesion images revealed the presence of individual casein micelles and larger clusters of aggregated casein micelles at MPC particle surfaces, whereas we observed no correlation between the solubility and averaged values of these nanomechanical properties. Furthermore, Derjaguin-Muller-Toropov modulus and adhesion images showed that the peripheral edges of individual casein micelles and their clusters had significantly higher values of the corresponding nanomechanical properties than other regions in the images, indicating the occurrence of the fusion of casein micelles. The surface area coverage or the percent area of the fused regions in an image revealed significant negative linear correlations with the solubility for both the control and calcium-depleted MPC. The present results support the hypothesis that the fusion of casein micelles at MPC powder particle surfaces is a causative factor for the solubility loss of MPC during storage and in turn suggest that the solubility loss may be alleviated by inhibiting the formation of a crust or skin on powder particle surfaces.

Correlation of the Abbe Number, the Refractive Index, and Glass Transition Temperature to the Degree of Polymerization of Norbornane in Polycarbonate Polymers.

The influences of the average degree of polymerization (Dp), which is derived from Mn and terminal end group, on optical and thermal properties of various refractive indexed transparent polymers were investigated. In this study, we selected the alicyclic compound, Dinorbornane dimethanol (DNDM) homo polymer, because it has been used as a representative monomer in low refractive index polymers for its unique properties. DNDM monomer has a stable amorphous phase and reacts like a polymer. Its unique reaction allows continuous investigation from monomer to polymer. For hydroxy end group and phenolic end group polymers, the refractive index (nd) decreased with increasing Dp, and both converged to same value in the high Dp region. However, the Abbe number (νd) of a hydroxy end group polymer is not dependent on Dp, and the νd of a phenolic end group polymer is greatly dependent on Dp. As for glass transition temperatures (Tg), both end group series were increased as Dp increased, and both converged to the same value.

Effects of soy protein isolate hydrolysates on cholecystokinin released by rat intestinal mucosal cells and food intake in rats.

Soy protein isolate hydrolysates (SPIH) were prepared from soy protein isolate (SPI). Effects of SPIH on a satiety signal cholecystokinin (CCK) and feeding behavior in rats were investigated. SPIH induced more CCK release (164.66 ± 2.40 pg/mL) by rat intestinal mucosal cells than SPI (143.33 ± 3.71 pg/mL). Meal size (MS), intermeal interval (IMI), and satiety ratio (SR = MS/IMI) of rats received different daily doses of SPIH or dietary fiber were detected for 40 days. A 100 mg/kg dose of SPIH resulted in a greater SR than an identical dose of dietary fiber, while a 300 mg/kg dose resulted in a less MS and IMI. A 500 mg/kg dose of SPIH had similar effects to the same dose of dietary fiber on reducing MS, extending IMI, and increasing SR, but resulted in a significantly less body weight at the end of the experiment (318.15 ± 17.83 g) than the dietary fiber group (340.28 ± 6.15 g).