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The introduction of varicella vaccines into routine pediatric immunization programs has led to a considerable reduction in varicella incidence. However, there have been reports of varicella, herpes zoster, and meningitis caused by the vaccine strain of varicella-zoster virus (VZV), raising concerns. Establishing the relationship between the wild-type and vaccine strains in VZV infections among previously vaccinated individuals is crucial. Differences in the single nucleotide polymorphisms (SNPs) among vaccine strains can be utilized to identify the strain. In this study, we employed nanopore sequencing to identify VZV strains and analyzed clinical samples. We retrospectively examined vesicle and cerebrospinal fluid samples from patients with VZV infections. One sample each of the wild-type and vaccine strains, previously identified using allelic discrimination real-time PCR and direct sequencing, served as controls. Ten samples with undetermined VZV strains were included. After DNA extraction, a long PCR targeting the VZV ORF62 region was executed. Nanopore sequencing identified SNPs, allowing discrimination between the vaccine and wild-type strains. Nanopore sequencing confirmed SNPs at previously reported sites (105,705, 106,262, 107,136, and 107,252), aiding in distinguishing between wild-type and vaccine strains. Among the ten unknown samples, nine were characterized as wild strains and one as a vaccine strain. Even in samples with low VZV DNA levels, nanopore sequencing was effective in strain identification. This study validates that nanopore sequencing is a reliable method for differentiating between the wild-type and vaccine strains of VZV. Its ability to produce long-read sequences is remarkable, allowing simultaneous confirmation of known SNPs and the detection of new mutations. Nanopore sequencing can serve as a valuable tool for the swift and precise identification of wild-type and vaccine strains and has potential applications in future VZV surveillance. [ABSTRACT FROM AUTHOR]

• Endomyocardial biopsy samples of acute myocarditis were analyzed by CIBERSORT. • Activated NK cells were the most prevalent population in two cases. • Activated memory CD4+ and M2 macrophages were the most prevalent in one case each. • Overall CIBERSORT results were consistent with those of immunohistochemistry. • CIBERSORT is a promising tool to analyze immune cell landscapes in myocarditis. Myocarditis is an inflammatory disease of the myocardium, which leads to cardiac dysfunction and heart failure. Previous studies have suggested that complex cross-talk between innate and adaptive immune responses is involved in the pathogenesis of acute myocarditis. Immunohistochemistry is the current standard method for the evaluation of infiltrating immune cells, however, it is difficult to investigate and quantify many immune cell populations using this technique. Endomyocardial biopsy samples of five pediatric patients with myocarditis were analyzed by cell-type identification by estimating relative subsets of RNA transcript (CIBERSORT), a computational method for quantifying cell fractions from tissue gene expression profiles. CIBERSORT results were then compared with immunohistochemistry analyses. Significant results of immune infiltrate deconvolution were obtained in four patients with fulminant myocarditis by CIBERSORT analysis. Among 22 immune cell types, 19 cell types were detected in one or more patients. Activated NK cells were the most prevalent population in two patients, whereas activated memory CD4+ T cells and M2 macrophages were the most prevalent population in one patient each. Overall CIBERSORT results were consistent with those of immunohistochemistry, although some discrepancies were observed. Infiltrating immune cell subsets detected by CIBERSORT analysis can reflect the time course of innate and adaptive immune responses in acute myocarditis. CIBERSORT may have the potential to characterize the detail of infiltrating immune cells in myocardial tissues and provide novel insights into the pathogenesis of acute myocarditis. [ABSTRACT FROM AUTHOR]

Congenital radioulnar synostosis (RUS) is a rare skeletal disorder that is characterized by fusion of the radius and ulna. As the etiology of RUS is largely unknown, its treatment options are currently limited. A de novo missense mutation in the zinc finger matrin-type 2 (ZMAT2) gene was newly identified in a 5-year-old boy with RUS using whole-exome sequencing. Herein, we sought to further explore the function of zmat2 in zebrafish. Whole-mount in situ hybridization revealed site-specific expression of zzmat2 in the pectoral fins (equivalent to human upper limbs) and craniofacial regions, while immunohistochemistry showed the expression of zZmat2 in the pectoral fins and heart region. Gene knockdown produced defects in the pectoral fins and dorso-ventral patterning. zzmat2 knockdown also caused embryo dorsalization, a phenotype consistent with reduced/insufficient bone morphogenetic protein (BMP) signaling. These abnormalities were partially rescued by zbmp2b RNA overexpression and fully rescued by simultaneous overexpression of wild-type zzmat2. Importantly, the overexpression of mutant zzmat2 corresponding to the newly-identified mutation did not fully rescue the dorso-ventral patterning defects. The above findings indicate that ZMAT2 regulates skeletal development via the BMP signaling pathway, and its mutation may lead to a loss or reduction in biological activity. Thus, the newly identified ZMAT2 mutation potentially plays a causal role in RUS through deregulation of BMP signaling. • Congenital radioulnar synostosis (RUS) is a rare skeletal disorder of largely unknown etiology. • We identify ZMAT2 as a potential disease-causing gene in a patient with RUS by whole-exome sequencing. • Zebrafish in vivo and ex vivo studies reveal that ZMAT2 can regulate skeletogenesis via BMP signaling. [ABSTRACT FROM AUTHOR]

Although associations among obesity, adipocytokines, and bone mineral density have been reported, the influence of adipocytokines on osteoporotic fractures remains unclear. This study aimed to assess the impact of the adipocytokines leptin and adiponectin on the risk of incident vertebral and long-bone fractures in postmenopausal women. Clinical data were obtained from the retrospective Nagano Cohort Study of outpatients followed at a single primary care institute in Nagano Prefecture, Japan, between 1993 and 2018. The primary outcome was the occurrence of incident vertebral or long-bone fractures. In total, 1167 Japanese postmenopausal women (mean age: 65.9 years) completed the follow-up and the average observation period was 7.2 years. The subjects were divided into 4 groups (quartile 1 to 4) based respective leptin and adiponectin values. Kaplan–Meier analysis demonstrated a significantly lower incident long-bone fracture rate in the higher quartiles of serum leptin levels (p = 0.002). A significantly higher and more rapid occurrence of incident vertebral fractures, but not long-bone fractures, was found in the highest adiponectin quartile (p < 0.001). A Cox proportional hazards model adjusted for confounders including age, body weight, and either leptin or adiponectin revealed lower leptin levels and higher adiponectin levels as significant independent risk factors for incident long-bone fractures (hazard ratio [HR] 0.70, 95% confidence interval [CI] 0.50–0.96; p = 0.03) and vertebral fractures (HR 1.18, 95% CI 1.02–1.37; p = 0.02), respectively. Therefore, serum leptin and adiponectin may be independent risk factors for osteoporotic fractures affecting different bone types and sites. Determining patient adipocytokine levels may help predict the occurrence of specific osteoporotic fractures, thereby enabling optimal treatment for osteoporosis and improving quality of life. • The influence of adipocytokines on osteoporotic fractures remains controversial. • Serum leptin and adiponectin are independent osteoporotic fracture risk factors. • Serum adipocytokine levels may predict the occurrence of osteoporotic fractures. [ABSTRACT FROM AUTHOR]

Background: Leukotriene (LT) B4 is a well-known inflammatory mediator and is implied to play some roles in glomerulonephritis. Although LTA4 hydrolase, a final-step key enzyme to produce LTB4, is located in glomerular mesangial cells, as well as in leukocytes, platelets, and endothelial cells, its precise distribution in the kidney other than in mesangial cells remains unknown. Therefore, we have investigated the localization of mRNA, protein, and enzyme activity of LTA4 hydrolase in the rat kidney.Methods: Microdissection reverse transcriptase-polymerase chain reaction was used for the determination of LTA4 hydrolase mRNA. The enzyme protein was detected by Western blot, and immunohistochemistry was performed. Finally, LTA4 hydrolase activity and LTB4 were assayed in kidney tissues.Results: LTA4 hydrolase mRNA was detectable in all microdissected nephron segments of the cortex and outer medulla. The corresponding size of approximately 70 kDa protein was shown in descending order in the inner medullary > outer medullary >/= cortical homogenates. The immunohistochemical study demonstrated the ubiquitous presence of the enzyme in all nephron segments of cortex, outer medulla, and inner collecting tubules. LTA4 hydrolase activity was detected in the inner medullary >/= outer medullary >/= cortical tissue homogenates. LTB4 was demonstrated in the inner medullary > outer medullary >/= cortical tissues during the basal condition, and was time-dependently increased by stimulation with arachidonic acid and ionomycin in the cytosolic fraction from outer medulla and in the glomerular suspension.Conclusions: These results strongly suggest that renal tubular cells as well as glomerular cells have an LTB4-forming potency, which may participate in physiological and pathophysiological roles in the kidney. [ABSTRACT FROM AUTHOR]