Sphingomyelin maintains the cutaneous barrier via regulation of the STAT3 pathway.

Epidermal tissues play vital roles in maintaining homeostasis and preventing the dysregulation of the cutaneous barrier. Sphingomyelin (SM), a sphingolipid synthesized by sphingomyelin synthase (SMS) 1 and 2, is involved in signal transduction via modulation of lipid-raft functions. Though the implications of SMS on inflammatory diseases have been reported, its role in dermatitis has not been clarified. In this study, we investigated the role of SM in the cutaneous barrier using a dermatitis model established by employing Sgms1 and 2 deficient mice. SM deficiency impaired the cutaneous inflammation and upregulated signal transducer and activator of transcription 3 (STAT3) phosphorylation in epithelial tissues. Furthermore, using mouse embryonic fibroblast cells, the sensitivity of STAT3 to Interleukin-6 stimulation was increased in Sgms-deficient cells. Using tofacitinib, a clinical JAK inhibitor, the study showed that SM deficiency might participate in STAT3 phosphorylation via JAK activation. Overall, these results demonstrate that SM is essential for maintaining the cutaneous barrier via the STAT3 pathway, suggesting SM could be a potential therapeutic target for dermatitis treatment.

Prediction Model with HLA-A*33:03 Reveals Number of Days to Develop Liver Cancer from Blood Test.

The development of liver cancer in patients with hepatitis B is a major problem, and several models have been reported to predict the development of liver cancer. However, no predictive model involving human genetic factors has been reported to date. For the items incorporated in the prediction model reported so far, we selected items that were significant in predicting liver carcinogenesis in Japanese patients with hepatitis B and constructed a prediction model of liver carcinogenesis by the Cox proportional hazard model with the addition of Human Leukocyte Antigen (HLA) genotypes. The model, which included four items-sex, age at the time of examination, alpha-fetoprotein level (log10AFP) and presence or absence of HLA-A*33:03-revealed an area under the receiver operating characteristic curve (AUROC) of 0.862 for HCC prediction within 1 year and an AUROC of 0.863 within 3 years. A 1000 repeated validation test resulted in a C-index of 0.75 or higher, or sensitivity of 0.70 or higher, indicating that this predictive model can distinguish those at high risk of developing liver cancer within a few years with high accuracy. The prediction model constructed in this study, which can distinguish between chronic hepatitis B patients who develop hepatocellular carcinoma (HCC) early and those who develop HCC late or not, is clinically meaningful.

Selective detection of urease-producing bacteria on the genital skin surface in patients with incontinence-associated dermatitis.

We aimed to investigate the association between the presence of cutaneous urease-producing bacteria and the development of incontinence-associated dermatitis (IAD) using an original urea agar medium as a step toward developing advanced preventive measures. In previous clinical assessments, we developed an original urea agar medium to detect urease-producing bacteria via the medium's colour changes. In a cross-sectional study, specimens were collected via the swabbing technique at genital skin sites in 52 stroke patients hospitalised in a university hospital. The primary objective was to compare the presence of urease-producing bacteria between the IAD and no-IAD groups. Determining the bacterial count was the secondary objective. The prevalence of IAD was 48%. A significantly higher detection rate of urease-producing bacteria was observed in the IAD group than in the no-IAD group (P = .002) despite the total number of bacteria being equivalent between them. In conclusion, we discovered that there was a significant association between the presence of urease-producing bacteria and IAD development in hospitalised stroke patients.

Bacterial species distribution on the genital skin of hospitalized patients with stroke manifesting incontinence-associated dermatitis: A cross-sectional study.

AIM: To compare the isolated and identified bacterial species colonizing on the genital skin between patients with and without incontinence-associated dermatitis. METHODS: This cross-sectional study included 102 patients with stroke admitted to an acute hospital in Japan. Swabs were collected, and bacterial species found in swabs were isolated and identified using a selective agar medium and simple identification kits. In addition to demographic information, severity of incontinence-associated dermatitis and the total bacterial counts were measured. RESULTS: Incontinence-associated dermatitis was present in 53.9% of the participants. Staphylococcus aureus was found in 50% of the participants with incontinence-associated dermatitis and only 17.9% of those without incontinence-associated dermatitis (P = 0.0029). Bacterial species distribution by erythema and skin erosion, which denote severity of incontinence-associated dermatitis, was different, but not significant; additionally, the total number of bacterial colonies was equivalent. CONCLUSIONS: Bacterial species distribution differed between patients with and without incontinence-associated dermatitis, whereas the total number of bacterial colonies was equivalent. A high detection rate of S. aureus on genital skin sites potentially affects the presence of incontinence-associated dermatitis and its severity. Geriatr Gerontol Int 2023; 23: 537-542.

Genetic association of IL17 and the importance of ABO blood group antigens in saliva to COVID-19.

The outbreak of COVID-19 caused by infection with SARS-CoV-2 virus has become a worldwide pandemic, and the number of patients presenting with respiratory failure is rapidly increasing in Japan. An international meta-analysis has been conducted to identify genetic factors associated with the onset and severity of COVID-19, but these factors have yet to be fully clarified. Here, we carried out genomic analysis based on a genome-wide association study (GWAS) in Japanese COVID-19 patients to determine whether genetic factors reported to be associated with the onset or severity of COVID-19 in the international meta-GWAS are replicated in the Japanese population, and whether new genetic factors exist. Although no significant genome-wide association was detected in the Japanese GWAS, an integrated analysis with the international meta-GWAS identified for the first time the involvement of the IL17A/IL17F gene in the severity of COVID-19. Among nine genes reported in the international meta-GWAS as genes involved in the onset of COVID-19, the association of FOXP4-AS1, ABO, and IFNAR2 genes was replicated in the Japanese population. Moreover, combined analysis of ABO and FUT2 genotypes revealed that the presence of oral AB antigens was significantly associated with the onset of COVID-19. FOXP4-AS1 and IFNAR2 were also significantly associated in the integrated analysis of the Japanese GWAS and international meta-GWAS when compared with severe COVID-19 cases and the general population. This made it clear that these two genes were also involved in not only the onset but also the severity of COVID-19. In particular, FOXP4-AS1 was not found to be associated with the severity of COVID-19 in the international meta-GWAS, but an integrated analysis with the Japanese GWAS revealed an association with severity. Individuals with the SNP risk allele found between IL17A and IL17F had significantly lower mRNA expression levels of IL17F, suggesting that activation of the innate immune response by IL17F may play an important role in the severity of SARS-CoV-2 infection.

Importance of HBsAg recognition by HLA molecules as revealed by responsiveness to different hepatitis B vaccines.

Hepatitis B (HB) vaccines (Heptavax-II and Bimmugen) designed based on HBV genotypes A and C are mainly used for vaccination against HB in Japan. To determine whether there are differences in the genetic background associated with vaccine responsiveness, genome-wide association studies were performed on 555 Heptavax-II and 1193 Bimmugen recipients. Further HLA imputation and detailed analysis of the association with HLA genes showed that two haplotypes, DRB1*13:02-DQB1*06:04 and DRB1*04:05-DQB1*04:01, were significantly associated in comparison with high-responders (HBsAb > 100 mIU/mL) for the two HB vaccines. In particular, HLA-DRB1*13:02-DQB1*06:04 haplotype is of great interest in the sense that it could only be detected by direct analysis of the high-responders in vaccination with Heptavax-II or Bimmugen. Compared with healthy controls, DRB1*13:02-DQB1*06:04 was significantly less frequent in high-responders when vaccinated with Heptavax-II, indicating that high antibody titers were less likely to be obtained with Heptavax-II. As Bimmugen and Heptavax-II tended to have high and low vaccine responses to DRB1*13:02, 15 residues were found in the Heptavax-II-derived antigenic peptide predicted to have the most unstable HLA-peptide binding. Further functional analysis of selected hepatitis B patients with HLA haplotypes identified in this study is expected to lead to an understanding of the mechanisms underlying liver disease.

Changes in activities of daily living, physical fitness, and depressive symptoms after six-month periodic well-rounded exercise programs for older adults living in nursing homes or special nursing facilities.

A 6-month, twice weekly, well-rounded exercise program (47 sessions in total) comprised of a combination of aerobic, resistance and flexibility training was provided for institutionalized older adults aged 60 to 93. We analyzed the data of 18 older adults who could stand and had attended more than 10% of the classes (mean participation rate: 54%) to examine changes in activities of daily living (ADL), physical fitness tests and depressive moods. The mean (+/- standard deviation, range) age of the participants was 71.3 (+/- 15.6, 60-93) in men and 85.9 (+/- 5.8, 72-93) in women. Significant improvement in ADL of the hand manipulation domain and borderline significant improvement in ADL of the mobility domain were observed (McNemar test p = 0.011 and 0.072, respectively). A 6-minute walk distance increased significantly from 151.6 m to 236.6 m (p = 0.01, paired t-test), and the result of the Soda Pop test, which tests hand-eye coordination, also improved significantly from 35.2 sec to 25.3 sec (p = 0.01, paired t-test). These findings suggest that such a program could be effective in improving the ADL and physical fitness of the elderly.

Synthetic lethality of the ALDH3A1 inhibitor dyclonine and xCT inhibitors in glutathione deficiency-resistant cancer cells.

The cystine-glutamate antiporter subunit xCT suppresses iron-dependent oxidative cell death (ferroptosis) and is therefore a promising target for cancer treatment. Given that cancer cells often show resistance to xCT inhibition resulting in glutathione (GSH) deficiency, however, we here performed a synthetic lethal screen of a drug library to identify agents that sensitize the GSH deficiency-resistant cancer cells to the xCT inhibitor sulfasalazine. This screen identified the oral anesthetic dyclonine which has been recently reported to act as a covalent inhibitor for aldehyde dehydrogenases (ALDHs). Treatment with dyclonine induced intracellular accumulation of the toxic aldehyde 4-hydroxynonenal in a cooperative manner with sulfasalazine. Sulfasalazine-resistant head and neck squamous cell carcinoma (HNSCC) cells were found to highly express ALDH3A1 and knockdown of ALDH3A1 rendered these cells sensitive to sulfasalazine. The combination of dyclonine and sulfasalazine cooperatively suppressed the growth of highly ALDH3A1-expressing HNSCC or gastric tumors that were resistant to sulfasalazine monotherapy. Our findings establish a rationale for application of dyclonine as a sensitizer to xCT-targeted cancer therapy.

Factors related to frequency of engaging in outside activities among elderly persons living an independent life at home.

We examined by cross-sectional design the association between the frequency of outside activity and the demographic, health problem, lifestyle, psychosocial and environmental factors among 239 community-dwelling elderly Japanese people (105 men and 134 women) aged 65 years and older, who lived independently at home. The associations of the outside activities more than 6-7 days per week based on a categorical questionnaire choice with potential factors were expressed by odds ratio (OR) and 95% confidence interval (CI) calculated through logistic regression analysis by sex. The proportions of those engaging in outside activity more than 6-7 days per week were 38.1% and 21.6% among men and women, respectively. The associations of the factors belonging to various fields with the frequency of engaging in outside activities were clearly pronounced among men. Elderly men engaging in outside activities more often were scored less for depression and more socially active than men engaging in outside activities less often. Multiple regression analysis revealed meeting and talking often with friend (OR=4.18, 95% CI: 1.06-16.5), current alcohol consumption (3.01, 1.06-8.54), having any hobby (3.59, 0.94-13.7), and easy access to public transportation (3.43, 1.28-9.16) as significant or borderline significant factors related to engaging in outside activities more than 6-7 days per week. Women who were currently employed engaged in outside activities more frequently. The frequency of engaging in outside activities was associated with factors belonging to various fields among elderly people living independently, particularly among men, suggesting its usefulness as an indicator of comprehensive well-being.

The EGF Receptor Promotes the Malignant Potential of Glioma by Regulating Amino Acid Transport System xc(-).

Extracellular free amino acids contribute to the interaction between a tumor and its microenvironment through effects on cellular metabolism and malignant behavior. System xc(-) is composed of xCT and CD98hc subunits and functions as a plasma membrane antiporter for the uptake of extracellular cystine in exchange for intracellular glutamate. Here, we show that the EGFR interacts with xCT and thereby promotes its cell surface expression and function in human glioma cells. EGFR-expressing glioma cells manifested both enhanced antioxidant capacity as a result of increased cystine uptake, as well as increased glutamate, which promotes matrix invasion. Imaging mass spectrometry also revealed that brain tumors formed in mice by human glioma cells stably overexpressing EGFR contained higher levels of reduced glutathione compared with those formed by parental cells. Targeted inhibition of xCT suppressed the EGFR-dependent enhancement of antioxidant capacity in glioma cells, as well as tumor growth and invasiveness. Our findings establish a new functional role for EGFR in promoting the malignant potential of glioma cells through interaction with xCT at the cell surface. Cancer Res; 76(10); 2954-63. ©2016 AACR.

Cloning of cDNAs encoding multiple subtypes of feline interferon-alpha from the feline epitherial cell line.

Mammalian interferon (IFN)-alpha consists of a 23-amino acid signal peptide and a 166-amino acid mature protein. Feline (Fe) IFN-alpha has an extra unique molecule consisting of a 171-amino acid mature protein with a 5-amino acid insertion. We cloned eight new subtypes of cDNA encoding FeIFN- alpha from a feline epithelial cell line. Among all the FeIFN-alpha subtypes, including six that have previously been reported, the variations were found to be far less than those of IFN-alphas of other animals.

[Association of flatfoot with pain, fatigue and obesity in Japanese over sixties].

PURPOSE: There is still uncertainty whether flatfoot, characterized by lower height of the medial longitudinal arch of the foot, is associated with foot symptoms in adults. Furthermore, few studies have been conducted to examine any association with obesity in older people. In this study, we therefore assessed the prevalence of flatfoot among community-dwelling older men and women using a footprint method, and analyzed relationships with foot symptoms and obesity. METHODS: The study sample consisted of 242 women and 98 men aged 60 years or older who were neither institutionalized nor disabled in activities of daily living. We collected footprints of both feet, data on weight and height, and information on foot symptoms and daily activity. From the footprints, we identified the H line by connecting the tip of the second toe and the intersection of the internal surface tangent of the foot and the external one. Then flatfoot was defined as concavity of the medial arch situated medial to the H line in either foot. Foot symptoms (pain and fatigue) were assessed using a 3-scale questionnaire: no pain (fatigue); present only when walking or standing; or present anytime. If present, the place of the pain or the fatigue was further identified. We analyzed associations with these symptoms and also with obesity assessed in terms of the body mass index (BMI). RESULTS: We identified flatfoot in 26.5% (95% Confidence Interval: 22.0-31.0) of the men and 25.7% (95% Confidence Interval: 20.2-31.2) of the women. In both sexes, the affected individuals had a higher prevalence of foot pain and fatigue than those unaffected. The association with the latter in women was particularly significant. There was a linear association between prevalence of flatfoot with the BMI category in women. In addition, the prevalence was lowest in the least obese category in men although this did not reach statistical significance. CONCLUSIONS: Flatfoot was significantly associated with the presence of pain and fatigue in women. Furthermore, a significant positive association with obesity was noted.

Stereocontrolled syntheses of alpha-C-mannosyltryptophan and its analogues.

The total synthesis of alpha-C-mannosyltryptophan (C-Man-Trp), a naturally occurring C-glycosylamino acid, was achieved from a commercially available alpha-methyl-D-mannoside in 10 steps including the following key steps: the C-glycosidation of a mannose derivative with a stannylacetylene, Castro indole synthesis, and Sc(ClO4)3-promoted coupling with L-serine-derived aziridine carboxylate. The glucose- and galactose-analogues of C-Man-Trp were also synthesized in a similar manner. Conformational analyses of the synthesized C-glycosyltryptophan and its synthetic intermediate are briefly discussed.

Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene.

We developed an electrochemical detection method for evaluating cellular physiological status based on the stringent response as a means to monitor cell viability. A reporter plasmid was constructed by inserting the beta-galactosidase gene (lacZ) under the control of the rpoS promoter, and then used to transform E. coli cells. Electrochemical responses from the products catalyzed by beta-galactosidase expressed by these E. coli cells were detected using the chronoamperometric technique in a nondestructive manner. Comparisons of response currents between the relA-positive strain and relA-negative strain revealed that increases in these currents were caused by the stringent response due to the stressful alcoholic environment, and thus as a model of stressful cultivating conditions. The current was proportional to the beta-galactosidase activity assayed by a conventional method that required the destruction of cells. The cellular physiological status, which depends on the stringent response as a viability marker, therefore, could then be evaluated online with a current using the rpoS-lacZ reporter gene in the relA-positive strain without pretreatment.

Long-term body weight fluctuation is associated with metabolic syndrome independent of current body mass index among Japanese men.

BACKGROUND: The relation between weight fluctuation and the risk of cardiovascular disease (CVD) is fairly consistent, although the physiologic basis for the relationship is uncertain. In the present study the association between long-term weight fluctuation and the development of metabolic syndrome (MS), a potent CVD risk factor, was investigated. METHODS AND RESULTS: A cross-sectional study of 664 Japanese men aged 40-49 years was conducted. The root mean square error around the slope of weight on age (weight - RMSE) was calculated by a simple linear regression model, in which the subject's actual weights at ages 20, 25, 30 years and 5 years prior to the study, as well as current weight, were dependent variables against the subject's age as the independent variable. Weight-RMSE was significantly and positively associated with the prevalence of each MS components (high blood pressure, hypertriglyceridemia, low-high density lipoprotein-cholesterol, high fasting glucose, and obesity). Such associations, as well as clustering of the MS component together with RMSE increase, were apparent among subjects with body mass index (BMI) <25 kg/m2, although the prevalence of MS or its components was much higher among overweight subjects (BMI >or=25 kg/m2). CONCLUSIONS: Development of MS possibly explains the risk of CVD not only in overweight or obese persons, but also in normal-weight persons with large weight fluctuation.

Association between serum leptin concentration and white blood cell count in middle-aged Japanese men and women.

BACKGROUND: Leptin's hematopoietic or proinflammatory role has been experimentally reported. We investigated whether serum leptin concentrations are associated with white blood cell (WBC) counts in humans. METHODS: Serum leptin concentrations of Japanese civil servants aged 40 to 59 years (1082 men and 200 women) were analyzed in relation to their WBC count. Serum leptin concentrations and WBC counts were measured by radioimmunoassay and automated particle counter respectively, using samples obtained at the time of the participants' annual health checkups. RESULTS: The geometric mean (+/-geometric standard deviation) leptin concentrations were 3.25 +/- 1.82 ng/mL and 6.25 +/- 3.99 ng/mL, and the geometric mean WBC counts, 5770 +/- 1269/mm(3) and 5107 +/- 1228/mm(3), in men and women respectively. The WBC count adjusted for age, body mass index (BMI), physical activity, and drinking and smoking habits increased together with the increase in leptin concentration. Multiple linear regression against WBC count by the leptin concentration and those covariates revealed a significant and independent association with serum leptin concentration especially in women (standardized beta = 0.31, p < 0.001), and also in men (standardized beta = 0.17, p < 0.001). BMI was not significantly associated with WBC counts in the multivariate model adjusting for leptin levels in both sexes. CONCLUSIONS: Our results are in line with leptin's hematopoietic or proinflammatory functions. The increased WBC counts often observed in obese people would be mediated by the increased leptin concentration.

Serum phospholipid transfer protein mass as a possible protective factor for coronary heart diseases.

BACKGROUND: Phospholipid transfer protein (PLTP) can generate pre-beta high-density lipoprotein (HDL), an efficient acceptor of peripheral cholesterol, by mediating a process called HDL conversion. The transfer of phospholipids to immature HDL is also essential in maintaining reverse cholesterol transport. The phospholipid transfer activity of PLTP has been associated with various patho-physiological conditions; however, little information is available concerning the relationship between PLTP mass and disease. METHODS AND RESULTS: Using a sandwich enzyme-linked immunosorbent assay, PLTP concentration was measured and related to the risk of developing cardiovascular disease in a worksite-based cohort of Japanese men (n=2,567). Multiple linear regression analysis showed significant associations between PLTP and HDL cholesterol, triglycerides, low-density lipoprotein cholesterol, and body mass index (standardized beta=0.395, -0.191, -0.064, and -0.064, respectively; R(2)=0.31). During the follow-up period, there were 10 cases of coronary heart disease (CHD) and 7 of stroke. The multivariate adjusted relative risk of CHD was 0.46 (95% confidence interval, 0.20-1.07) for an increase of 1 standard deviation in the PLTP value (p=0.071). PLTP concentration was not related to the risk of stroke. CONCLUSIONS: The results of this prospective study indicate that the serum PLTP concentration would serve as a predictor of CHD, independent of HDL cholesterol, triglycerides and other established risk factors.

Association of white blood cell count and clustered components of metabolic syndrome in Japanese men.

BACKGROUND: The role of inflammation in the genesis of cardiovascular disease has attracted attention and in the present study the association among metabolic syndrome (MS), white blood cell (WBC) count, and insulin concentration was investigated. METHODS AND RESULTS: A cross-sectional study of 3,594 Japanese men aged 34-69 years evaluated the MS components (high blood pressure, hypo-high density lipoprotein (HDL)-cholesterolemia, hypertriglyceridemia, hyperglycemia), as defined by the criteria given in the Third Report of the National Cholesterol Education Program Expert Panel on Detection Evaluation, and Treatment of High Blood Cholesterol in Adults, except for obesity [body mass index (BMI) >/=25 kg/m(2)]. WBC count had a positive correlation with BMI, blood pressure, triglyceride, glucose and insulin, and a negative correlation with HDL-cholesterol. The multi-adjusted means of WBC count and insulin concentration were significantly higher in MS subjects defined as having 3 or more of the components than in non-MS subjects with no more than 2 components. Both means also increased with the number of MS components (p<0.001 for trend). In the multiple linear regression analysis, BMI, HDL-cholesterol, systolic blood pressure, glucose and triglyceride had a significant and independent association with WBC count, but the insulin concentration did not. CONCLUSIONS: The cluster of MS components based on insulin resistance may cause low-grade inflammation.

Reproducibility and validity of a simple checklist-type questionnaire for food intake and dietary behavior.

BACKGROUND: A simple, reliable, and valid food questionnaire is needed in clinical dietary assessments, community health education, and multi-purpose epidemiologic studies to obtain a crude measure of dietary intake. METHODS: To assess the validity and reproducibility of a simple 4-point scale food intake and behavior checklist, it was compared to two 3-day weighed dietary records. The FBC was administered to 47 students of a dietician course and their parents (n = 94) over a 9-month interval to assess the reproducibility. The mean intakes of selected food groups assessed by the two dietary records completed between food intake and behavior checklists were compared to the responses to the food intake and behavior checklist to assess its validity. RESULTS: The kappa statistics for reproducibility ranged from 0.25 for confectionaries to 0.63 for a preference for fatty foods (median, 0.39). There was a reasonable level of correlation between the dietary record and the food intake and behavior checklist in the intake of eggs, milk, and fruits (r = 0.53, 0.56, and 0.50, respectively). There was a weaker but still significant correlation in the intake of vegetables, and alcohol (r = 0.31 and 0.45, respectively). No significant correlation was observed in the intake of meat, fish, confectionaries, and soft drinks. However, those who reported consuming mainly fish rather than meat were found to eat significantly less meat and animal fat. Similarly, those who did not prefer fatty foods consumed significantly less meat, animal fat, and polyunsaturated fatty acids. CONCLUSIONS: This simple food checklist was useful in collecting data on egg, milk, and fruit consumption. Assessing intake frequency of vegetables, meat or fish with the FBC may be useful in screening high- or low-intake individuals.