Systemic immune responses do not affect significant immune responses in the skin.

Fish skin plays an important role in defending against pathogens in water, primarily through the secretion of skin mucus containing various immune-related factors. Local immune responses in the skin activate systemic immune responses by inflammatory cytokines. However, it remains unclear whether immune responses in the skin occur after systemic immune responses caused by pathogen invasion into the fish body. This study aimed to clarify the relationship between systemic immune responses and skin responses after intraperitoneal injection of formalin-killed cells (FKC) of Vibrio anguillarum. Although systemic inflammatory responses were observed in the spleen after injection, expression changes in the skin did not show significant differences. In contrast, expression of hemoglobin subunit genes significantly increased in the skin after FKC injection, suggesting that erythrocytes infiltrate extravascularly.

[A Trial of Practical Training on Online Medication Instructions in Clinical Preparatory Education: Development of Human Resources to Provide New Forms of Medical Care after the COVID-19 Pandemic].

The coronavirus disease 2019 (COVID-19) pandemic has considerably affected several social services. The Ministry of Health, Labour, and Welfare has partially revised the Pharmaceuticals and Medical Devices Law and established legislations on permanent online medication instructions. Based on these social needs, the development of human resources to provide online medication instructions is vital. Therefore, we developed a training program for providing online medication instructions in preparatory clinical education. Pharmacy students who had conducted medical interviews with standardized patients participated in the training. Educational outcomes were evaluated using an objective multiple-choice test and free description before and after practical training. The median number of correct answers on objective tests on the legislation on online medication instructions increased significantly. Based on the free description analysis, students were able to comprehend the influence of communication environment on the quality of medication instructions. Based on the results of the direct evaluation using objective testing and indirect evaluation by analyzing the free descriptions, they also acquired the skills necessary for providing online medication instructions. Therefore, this training program can contribute to mastering the provision of online medication instructions.

Association between birth weight and prevalence of cardiovascular disease and other lifestyle-related diseases among Japanese population: JPHC-NEXT Study.

BACKGROUND: An association between birth weight and cardiovascular disease (CVD) in adulthood has been observed in many countries; however, only a few studies have been conducted in Asian populations. METHODS: We used data from the baseline survey (2011-2016) of the Japan Public Health Center-based Prospective Study for the Next Generation Cohort, which included 114,105 participants aged 40-74 years. Adjusted prevalence ratios (aPRs) and 95% confidence intervals (CIs) were calculated from the prevalence of present and past histories of CVD and other lifestyle-related diseases, including hypertension, diabetes, hyperlipidemia, and gout, by birth weight, using Poisson regression. RESULTS: The prevalence of CVD increased with lower birth weight, with the highest prevalence among those with birth weight under 1,500 grams (males 4.6%; females 1.7%) and the lowest one among those with birth weight at or over 4,000g (males 3.7%: females 0.8%). Among 88.653 participants (41,156 males and 47,497 females) with complete data on possible confounders, birth weight under 1,500g was associated with a higher prevalence of CVD (aPR 1.76 [95%CI 1.37-2.26]), hypertension (aPR 1.29 [95%CI 1.17-1.42]), and diabetes (aPR 1.53 [95%CI 1.26-1.86]) when a birth weight of 3,000-3,999 grams was used as the reference. Weaker associations were observed for birth weight of 1500-2499 grams and 2500-2999 grams, while no significant associations were observed for birth weight at or over 4000 g. The association between birth weight and the prevalence of hyperlipidemia was less profound, and no significant association was observed between birth weight and gout. CONCLUSION: Lower birth weight was associated with a higher prevalence of CVD, hypertension, and diabetes in the Japanese population.

Association Between Skin Autofluorescence and White Blood Cell Counts in Older Adults.

BACKGROUND/AIM: Advanced glycation end products (AGEs) accumulate in the body with increasing age. However, their excessive accumulation may lead to various inflammatory and chronic diseases. While it is common for older adults to experience various comorbidities, there is a scarcity of published literature documenting the specific impact of ageing and comorbidities on AGEs in this population. The present study aimed to retrospectively evaluate the correlation among AGEs in the skin, calendar age, and comorbidities in older adults. PATIENTS AND METHODS: Accumulated AGEs in the skin were assessed by non-invasive measurement of skin autofluorescence (SAF) inside the forearm. This retrospective study included individuals who underwent SAF measurements at Shujitsu University Community Pharmacy with or without a prescription from October 2019 to October 2021. Subsequently, the associations between SAF, calendar age, comorbidities, and blood test parameters were investigated. RESULTS: SAF showed a positive correlation with calendar age for all enrolled participants; the correlation weakened for participants aged ≥50 years and plateaued for those aged ≥60 years. Furthermore, we observed a significant increase in SAF among all participants with comorbidities compared to those without comorbidities. By contrast, among participants aged ≥50 years, SAF did not show a significant association with comorbidities. However, SAF was significantly positively correlated with white blood cell (WBC) counts in these aged populations. CONCLUSION: The non-invasive assessment of SAF holds promise in evaluating changes in the physical condition associated with WBC counts among older adults.

Analysis of the distribution of adult height standard deviation scores in relation to prepubertal height standard deviation scores using longitudinal growth data: -Investigation of the catch-up rates of children with short stature to attain normal adult height.

Using the longitudinal growth data of 13,809 individuals in the Akita Prefecture, the percentage distributions of their adult height (AH) standard deviation scores (SDS) in relation to their prepubertal height SDS were obtained. The AH SDS increased with negative prepubertal height SDS and decreased with positive prepubertal height SDS, showing that a greater amount of change was associated with a greater interval of the prepubertal height SDS from the mean. The proportions of individuals who achieved normal AH stratified by prepubertal height SDS were as follows: 67.1%, in the group with prepubertal height SDS of -2.5 < to ≤ -2.0 SD, 46.0% in the group with -3.0 < to ≤ -2.5 SD, 75.2% in the group with +2.0 ≤ to < +2.5 SD, and 55.1% in the group with +2.5 ≤ to < +3.0 SD. Of all participants with short stature at prepuberty, 58.4%, 33.8%, 8.3%, and 0% of those with prepubertal height SDS of ≤ -2.0 SD, ≤ -2.5 SD, ≤ -3.0 SD, and ≤ -3.5 SD attained normal AH, respectively. On average, it is difficult for children with prepubertal height SDS of ≤ -2.5 SD to attain normal AH.

Type I interferon induced by polyinosinic-polycytidylic acid does not contribute to the efficacy of a formalin-killed cell vaccine against Edwardsiella piscicida in the Japanese flounder (Paralichthys olivaceus).

Polyinosinic-polycytidylic acid (poly I:C) is a type of pathogen-associated molecular pattern that can strongly induce the expression of type I interferon (I-IFN). Our previous study has demonstrated that the combination of poly I:C with a recombinant protein antigen not only stimulated the expression of I-IFN but also conferred protection against Edwardsiella piscicida in the Japanese flounder (Paralichthys olivaceus). In this study, our aim was to develop a better immunogenic and protective fish vaccine, for which we intraperitoneally coinjected P. olivaceus with poly I:C and formalin-killed cells (FKCs) of E. piscicida and compared the efficiency of protection against E. piscicida infection with that of FKC vaccine alone. Results showed that the expression levels of I-IFN, IFN-γ, interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, and the interferon-stimulated genes (ISGs) ISG15 and Mx were significantly increased in the spleen of fish inoculated with poly I:C + FKC. The results of ELISA showed that the levels of specific serum antibodies in the FKC and FKC + poly I:C groups were gradually increased until 28 days postvaccination and were significantly higher than those in the PBS and poly I:C groups. At 3 weeks after vaccination in the challenge test, the respective cumulative mortality rates of fish in the PBS, FKC, poly I:C, and poly I:C + FKC groups were 46.7%, 20.0%, 33.3%, and 13.3% under low-concentration challenge and 93.3%, 46.7%, 78.6%, and 53.3% under high-concentration challenge. This study showed that poly I:C may not provide an effective adjuvant effect with FKC vaccine for intracellular bacterial infections.

Whole-genome analysis of red sea bream iridovirus spread in 2021 in Japan provided epidemiological and viral traits insight.

Red sea bream iridovirus (RSIV) is the pathogen that causes red sea bream iridoviral disease. It causes a huge loss to the Japanese aquaculture industry. In 2021, outbreaks of red sea bream iridovirus occurred in South Japan. This study analysed nine whole-genome sequences of RSIV isolated in Oita and Ehime Prefectures in 2021 using a short-read next-generation sequencer. Nine isolates had highly uniform sequences, and there was no variant depending on locations or host species. Phylogenetic analyses with other reported megalocytivirus isolates showed that RSIV isolated in 2021 was genetically different from RSIV previously isolated in Oita and Ehime Prefectures in 2017-2019. These results suggest that RSIV isolated in Oita and Ehime Prefectures in 2021 might spread from a common ancestor different from the recent one. Additionally, it was found that RSIV isolated in 2021 had sequence mutations on protein-coding sequences that may be involved in viral pathogenicity and infectivity.

Preschool-children's height, trend, and causes: Japanese national surveys 1990-2010.

We observed trends in the height of children aged 3 to 6 in Japan using data from the National Growth Survey on Preschool Children in the years 1990, 2000, and 2010. Average standard deviation (SD) scores of height decreased from 0.39 (SD 1.02) in 1990 (n = 3,684) to 0.37 (SD 1.05) in 2000 (n = 2,981) and 0.33 (SD 1.07) in 2010 (n = 2,027). Mothers of children in later waves were taller, older, and more likely to be primiparous; children in later waves had shorter gestational age, lower birth weight, and were less likely to have been fed less with formula or solid foods before 6 mo. The only factor that consistently contributed to a reduction in children's height for both 1990-2000 and 2000-2010 was a reduction in birthweight SD score (indirect effect on height -1.5 [95% CI: -1.9, -1.1] mm for 1990-2000 and -1.2 [95% CI: -1.8, -0.8] mm for 2000-2010). Factors that contributed, although not significantly or consistently between the two periods, were changes in pre-pregnancy BMI, smoking during pregnancy, multiple pregnancies, gestational age, BMI at birth, and use of formula and solid foods before 6 mo. Secular increases in maternal age, height, and primiparity contributed to increasing children's height.

Secular trends in early motor development between 1980 and 2010 in Japan.

OBJECTIVE: Recent changes in birth characteristics in Japan may have a potential influence on children's developments. Therefore, we investigated secular trends in gross motor milestones. DESIGN: Data were collected from an official Japanese nationwide serial cross-sectional survey conducted every 10 years since 1960. 22 320 participants aged 2-18 months were identified from the four surveys from 1980 to 2010. OUTCOMES: We assessed whether or not a child achieved four gross motor milestones including rolling over (rolling), sitting without support (sitting), standing with support (standing) and walking alone (walking). The target age was defined as the age when the attainment rate ranged from >5% to >95% of the total. Multivariate logistic regression models were fitted. RESULTS: The final cohort included 20 570 children. The target ages were determined as follows: 3-6 months for rolling; 5-9 months for sitting; 6-11 months for standing; and 9-15 months for walking. The attainment rates of sitting, standing and walking in 1990 were higher than those in 2010, even after adjusting for child characteristics (sitting: adjusted OR (aOR)=2.07 (95% CI 1.62 to 2.65); standing: aOR=1.63 (95% CI 1.32 to 2.02); and walking: aOR=1.61 (95% CI 1.34 to 1.95)). CONCLUSIONS: The proportion of children who attained three motor milestones (sitting, standing and walking) by set target ages decreased between 1990 and 2010. The contribution of birth characteristics including a decrease in gestational age and fetal growth, as well as changes in other child characteristics, failed to explain why this decrease occurred.

Development of single nucleotide polymorphism (SNP) application for detection and genotyping of RSIV-type megalocytiviruses.

Red sea bream iridovirus (RSIV) belonging to the genus Megalocytivirus of the family Iridoviridae is the cause of serious mass mortality of cultured marine fishes. RSIV-type megalocytiviruses show extremely high nucleotide sequence identities. Thus, epidemiological studies on this virus are limited. This study developed two primer sets amplifying the regions possessing single nucleotide polymorphism (SNP) to determine the relationships and divergence of RSIV-type megalocytiviruses isolated from cultured marine fishes in Japan. The two regions were designed according to the genome sequences of the representative RSIV genotype II of megalocytivirus members in GenBank. The SNP 1 and 2 regions have sequences homologous to hypothetical protein ORF 24 and ORF 31, respectively, of RSIV (accession no. AP017456.1). By sequencing the regions, 53 polymorphic sites were identified. The phylogenetic analysis of 25 RSIV-type megalocytivirus isolates, classified into RSIV cluster, was clustered into eight haplotypes (seven haplotypes from Oita, two haplotypes from Ehime, and one haplotype shared between Oita and Ehime). These findings suggested that SNP in the RSIV genome is a powerful application for the detection and identification of RSIV-type megalocytiviruses.

Reduced pubertal growth in children with obesity regardless of pubertal timing.

Childhood obesity affects both pubertal growth and pubertal timing. We evaluated pubertal timing-mediated effects of childhood obesity on pubertal growth. This retrospective, representative-population-based cohort study included 6,733 boys and 6,916 girls born between April 1975 and March 1976 in Akita Prefecture, Japan. Individual changes in height standard deviation score between 7 and 17 years (ΔHtSDS), body mass index Z-score at 7 years (BMIZ), and estimated age at peak height velocity (ÂPHV) were used as surrogate indicators of pubertal growth, childhood obesity and pubertal timing, respectively. ÂPHV-mediated effect of BMIZ on ΔHtSDS was evaluated, and non-ÂPHV-mediated effect was calculated. Based on BMIZ, participants were categorized into three groups (underweight, normal-weight and obese groups), and the differences in ΔHtSDS between obese and normal-weight or underweight groups and ratios of non-ÂPHV-mediated effect were determined. ΔHtSDS values in the obese group were lower by 1.23 in boys and 1.17 in girls than those in the underweight group and by 0.87 in boys and 0.85 in girls than those in the normal-weight group. Non-ÂPHV-mediated effect on the reduced ΔHtSDS in the obese group compared to the underweight and normal-weight groups accounted for 68% and 71% in boys and 59% and 64% in girls, respectively. Thus, childhood obesity is associated with reduced pubertal growth regardless of pubertal timing. This reduced pubertal growth observed in children with obesity could be more affected by non-pubertal timing-mediated effect rather than pubertal timing-mediated effect.

Microdeletion in Xq28 with a polymorphic inversion in a patient with FLNA-associated progressive lung disease.

Lung phenotype was reported as a novel phenotype in patients with mutations in the filamin A gene (FLNA) in 2011. FLNA mutations can result in pulmonary hyperinflation during the neonatal period or early infancy with progressive respiratory failure, culminating in a diagnosis of FLNA-associated progressive lung disease, particularly if the patient has periventricular nodular heterotopia and cardiac complications, such as patent ductus arteriosus, atrial septal defect, and pulmonary hypertension. We report the first Japanese case of FLNA-associated progressive lung disease caused by a microdeletion in Xq28 encompassing the FLNA gene with a polymorphic inversion.

Establishment of a longitudinal growth chart corresponding to pubertal timing.

A standard growth chart is indispensable for evaluating an individual's growth. In Japan, the cross-sectional growth chart from fiscal year 2000 is most commonly used in the clinical setting. However, when using the current standard growth chart to assess growth during puberty, two problems are encountered. First, the individual pubertal height trajectory does not fit the cross-sectional growth chart because the pubertal height curve of individuals rises more sharply than that indicated by the cross-sectional growth chart. Second, variations in the timing of an individuals' growth spurt render it difficult or impossible to assess individual growth patterns using a single chart. To address these two issues, new growth charts were established using height measurements of 6744 boys and 6929 girls born between April 1975 and March 1976 in the Akita Prefecture. Individuals whose age at peak height velocity (agePHV) was 2 standard deviation greater or lesser than the mean were excluded, and the remaining participants were divided into three groups according to the first and third quartiles of agePHV. Finally, we established three longitudinal growth charts each for boys and girls based on a healthy Japanese population.

Complete Kisspeptin Receptor Inactivation Does Not Impede Exogenous GnRH-Induced LH Surge in Humans.

Context: Mutations in the kisspeptin receptor (KISS1R) gene have been reported in a few patients with normosmic congenital hypogonadotropic hypogonadism (nCHH) (OMIM #146110). Objectives: To describe a female patient with nCHH and a novel homozygous KISS1R mutation and to assess the role of kisspeptin pathway to induce an ovulation by GnRH pulse therapy. Design, Setting, and Intervention: Observational study of a patient including genetic and kisspeptin receptor functions and treatment efficiency using a GnRH pump. Main Outcome Measure: Response to pulsatile GnRH therapy. Results: A partial isolated gonadotropic deficiency was diagnosed in a 28-year-old woman with primary amenorrhea and no breast development. A novel homozygous c.953T>C variant was identified in KISS1R. This mutation led to substitution of leucine 318 for proline (p.Leu318Pro) in the seventh transmembrane domain of KISS1R. Signaling via the mutated receptor was profoundly impaired in HEK293-transfected cells. The mutated receptor was not detected on the membrane of HEK293-transfected cells. After several pulsatile GnRH therapy cycles, an LH surge with ovulation and pregnancy was obtained. Conclusion: GnRH pulsatile therapy can induce an LH surge in a woman with a mutated KISS1R, which was previously thought to be completely inactivated in vivo.

Mismatch between fetal sexing and birth phenotype: a case of complete androgen insensitivity syndrome.

With advancing maternal age, the number of prenatal genetic tests is increasing in many countries. Prenatal genetic tests, such as amniocentesis, chorionic villus sampling and non-invasive prenatal testing, can disclose fetal chromosomal sex, although these tests were originally designed to prenatally diagnose chromosomal aneuploidies, such as trisomy 21, 18 and 13. Complete androgen insensitivity syndrome (CAIS) is an X-linked recessive disorder caused by an androgen receptor dysfunction leading to hormone resistance. The affected individuals are genetic males as shown by 46,XY but present complete female external genitalia and normal breast development at puberty albeit without menstruation. CAIS is commonly diagnosed in adolescence based on primary amenorrhea or in childhood based on inguinal hernia or testis-like masses in the inguinal region. In the present report, we describe a baby in whom CAIS was diagnosed immediately after birth based on a mismatch between the fetal karyotype detected by amniocentesis and the external genitalia phenotype at birth. We speculate that the increase in the number of prenatal genetic tests is contributing to the early detection of 46,XY disorders of sex development, especially those previously called complete sex reversal, which is supposedly diagnosed during childhood or adolescence. Hence, it is necessary to understand the disease-specific hormone profile at each developmental stage for accurate diagnosis.

Ecological analysis of secular trends in low birth weight births and adult height in Japan.

BACKGROUND: Japan, which currently maintains the highest life expectancy in the world and has experienced an impressive gain in adult height over the past century, has suffered a dramatic twofold increase in low birth weight (LBW) births since the 1970s. METHODS: We observed secular trends in birth characteristics using 64 115 249 live births included the vital statistics (1969-2014), as well as trends in average height among 3 145 521 adults born between 1969 and 1996, included in 79 surveys conducted among a national, subnational or community population in Japan. RESULTS: LBW rates exhibited a U-shaped pattern showing reductions until 1978-1979 (5.5%), after which it increased. Conversely, average adult height peaked for those born during the same period (men, 171.5 cm; women, 158.5 cm), followed by a reduction over the next 20 years. LBW rate and adult height showed a strong inverse correlation (men, r=-0.98; women, r=-0.88). A prediction model based on birth and economical characteristics estimated the national average of adult height would continue to decline, to 170.0cm (95% CI 169.6 to 170.3) for men and 157.9cm (95% CI 157.5 to 158.3) for women among those born in 2014. CONCLUSIONS: Adult height in Japan has started to decline for those born after 1980, a trend that may be attributed to increases in LBW births over time. Considering the known association between shorter adult height and adverse health outcomes, evidence of population-level decline in adult health due to long-term consequences of increasing LBW births in Japan is anticipated.

Portosystemic shunt as a cause of congenital hyperinsulinemic hypoglycemia.

Congenital hyperinsulinemic hypoglycemia (CHH) is characterized by the inappropriate secretion of insulin from pancreatic beta cells in the presence of hypoglycemia. We herein describe the case of a 5-month-old boy with CHH due to congenital portosystemic shunt (CPSS). Insulin secreted from pancreatic beta cells flows into the portal vein and is first metabolized in the liver. First-pass elimination of insulin in the liver leads to great decrease in insulin concentration by approximately 40-80% in humans. CPSS accounts for a large quantity of insulin delivery into the systemic circulation due to the lack of hepatic first-pass elimination. Hypoglycemia can result from consistently high levels of insulin after reaching normal glucose level. CPSS therefore should be considered as a rare cause of CHH, especially in the case of post-prandial hyperinsulinemic hypoglycemia.

In vitro characteristics of cyprinid herpesvirus 2: effect of kidney extract supplementation on growth.

Herpesviral hematopoietic necrosis caused by goldfish hematopoietic necrosis virus (now identified as cyprinid herpesvirus 2, CyHV-2) has contributed to economic losses in goldfish Carassius auratus culture and is becoming a major obstacle in Prussian carp C. gibelio aquaculture in China. Several reports have described difficulties in culturing the virus, with the total loss of infectivity within several passages in cell culture. We succeeded in propagating CyHV-2 with a high infectious titer in a RyuF-2 cell line newly derived from the fin of the Ryukin goldfish variety using culture medium supplemented with 0.2% healthy goldfish kidney extract. The addition of kidney extract to the medium enabled rapid virus growth, resulting in the completion of cytopathic effect (CPE) within 4 to 6 d at 25°C. The extract also enabled reproducible virus culture with a titer of 105-6 TCID50 ml-1. The virus cultured using this protocol showed pathogenicity in goldfish after intraperitoneal injection. The virus grew in RyuF-2 cells at 15, 20, 25, 30, and 32°C but not at 34°C or higher. Higher incubation temperatures allowed earlier development of CPE, but culture at 30 and 32°C yielded a lower virus titer than that obtained at other temperatures because of heat inactivation of the propagated virus during cultivation. Cell lines derived from goldfish and ginbuna C. langsdorfii showed high susceptibility to the virus; cell lines from carp were susceptible to the virus using a medium containing goldfish kidney extract, but EPC, FHM, and BF-2 cell lines did not produce any CPE, even in the presence of the extract.

Clinical manifestations of Xq28 functional disomy involving MECP2 in one female and two male patients.

Subtelomeric imbalances are a frequent cause of cytogenetic abnormalities in patients with unexplained intellectual disability. Functional disomy of Xq28 involving the methyl-CpG-binding protein 2 gene (MECP2) has been observed mostly in subtelomeric duplications. We identified three patients with functional disomy of Xq28. A female patient showed an unbalanced translocation between 12q24.33 and Xq28. Two male patients showed an unbalanced translocation between Xq27.1- Yq11.22 and a recombinant X-chromosome containing duplicated material from Xq27.1 on Xp telomere, respectively. All three patients exhibited severe developmental delay, hypotonia, seizures, and distinctive facial features, including flat nasal bridge and hypertelorism. Additionally, brain magnetic resonance imaging (MRI) showed characteristic findings in each patient, including frontal dominant brain atrophy and hypoplasia of the corpus callosum, which are common findings in patients with functional disomies of Xq28 and interstitial duplications of Xq28, including MECP2. Brain MRI revealed a cystic lesion in the periventricular white matter in a patient, similar to our previous experience in patients with MECP2 duplication syndrome. Thus, white matter abnormalities may frequently be seen in cases of patients with additional MECP2 copies. © 2013 Wiley Periodicals, Inc.

Living donor liver transplantation for glycogen storage disease type Ib.

Glycogen storage disease type 1b (GSD-1b) is due to an autosomal recessive inborn error of carbohydrate metabolism caused by defects in glucose-6-phosphatase translocase. Patients with GSD-1b have severe hypoglycemia with several clinical manifestations of hepatomegaly, obesity, a doll-like face, and neutropenia. Liver transplantation has been indicated for severe glucose intolerance. This study retrospectively reviewed 4 children with a diagnosis of GSD-1b who underwent living-donor liver transplantation (LDLT). Between November 2005 and June 2008, 96 children underwent LDLT with overall patient and graft survival of 92.3%. Of these, 4 (4.2%) were indicated for GSD-1b. All patients are doing well with an excellent quality of life because of the stabilization of glucose intolerance, decreased hospital admission, and normalized neutrophil count. LDLT appears to be a feasible option and is associated with a better quality of life for patients with GSD-1b. Long-term observation may be necessary to collect sufficient data to confirm the efficacy of this treatment modality.