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Poor oral health has been linked to an increased risk of mortality. People with good cooking skills may be able to cope with problems caused by oral dysfunction, such as restricted food choice. This study investigated whether cooking skills modified the association between oral health status and mortality. A 3-year follow-up was conducted among participants of the Japan Gerontological Evaluation Study, a population-based cohort study of Japanese adults aged 65-101 years. Oral health assessment comprised number of teeth and three oral functions (awareness of swallowing difficulty, decline in masticatory function, and dry mouth). Cooking skills were assessed using a valid scale adapted for Japanese participants. Mortality from 2016 to 2019 was analysed for 10,121 respondents. Cox regression models were used to estimate hazard ratios for mortality risk adjusting for potential confounders, including sociodemographic factors and health status. During the follow-up, there were 488 deaths (4.8% of all participants). Both fewer teeth and poor oral function were associated with a greater mortality risk. Cooking skills modified the association only for oral function. Stratification by cooking skill score showed that the hazard ratios for ≥2 oral difficulties (vs. none) was 2.06 (95% confidence interval: 1.43-2.96) among those with low cooking skills and 1.25 (95% confidence interval: 0.92-1.71) among those with high cooking skills after adjusting for potential confounders. Improving cooking skills may be key to mitigating health risks owing to poor oral functions., (© The Author(s) 2023. Published by Oxford University Press on behalf of the British Geriatrics Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Painful ganglion cysts that develop in the hallux and finger usually enlarge progressively to the peripheral direction. Simple resection of satellite ganglion cyst alone has been reported to cause a high rate of recurrence and treatment is often very difficult. The purpose of this study is to evaluate the appropriate surgical treatment for painful satellite ganglion cysts in the hallux and finger and discuss the origin of the ganglion cysts in cases treated surgically at our hospital. We reviewed five cases (three males and two females, ages 55-87 years), three of which occurred in the hallux and two in the finger. In all cases, the preoperative magnetic resonance image showed a large fluid of the flexor tendon sheath. And also, joint effusion was found in the metatarsophalangeal joint and the proximal interphalangeal joint. The first case of the hallux ganglion underwent simple excision of the cyst and had recurrences three times. In the other four cases, the additional synovectomy of the metatarsophalangeal joint and the proximal interphalangeal joint was performed along with ganglion cyst excision. These cases had no recurrence up to 1 year after operation. Recently, there have been reports that tendon sheath ganglions are connected to the ankle, wrist, hallux, and phalangeal joints. Although there are a few cases in our department, satellite ganglion cyst of the hallux and finger possibly originates from adjacent joints. Additional synovectomy of the affected joint should be performed for the excision of satellite ganglion cyst to prevent recurrence., (© Japan College of Rheumatology 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objectives: Immunosuppressive therapy is the mainstay of treatment for child-onset systemic lupus erythematosus (cSLE). Since epidemiological data on Japanese cSLE patients are not available, we evaluated the trends in how treatment choices have changed over time in Japan., Methods: Using the Japanese health insurance database provided by Medical Data Vision Co., Ltd, we identified cSLE patients and evaluated changes in the use of corticosteroids and immunosuppressive medications and maximum daily doses of prednisolone from 2009 to 2018., Results: Of 182 cSLE patients, 86% were female, and the median age was 14 years. Oral prednisolone was used in more than 97% of cSLE patients during the study period, and the median of the maximum daily dose in each patient decreased over time. Intravenous cyclophosphamide was used less frequently after 2016, while mycophenolate mofetil and hydroxychloroquine were used frequently after 2016. The use of mizoribine reduced after 2014, whereas the other immunosuppressive medications showed no significant change over time; the use of biological agents was very limited., Conclusions: Oral prednisolone was the mainstay of treatment for cSLE, and the maximum daily dose has reduced over the past decade. The most frequently prescribed immunosuppressive therapy has shifted to mycophenolate mofetil over time., (© Japan College of Rheumatology 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: the present study aimed to investigate the relationship between the frequency of eating together and the risk of weight loss in older adults., Methods: this was a three-year follow-up prospective cohort study based on a self-reported questionnaire. We used data from the Japan Gerontological Evaluation Study (JAGES) conducted in 2016 and 2019. The participants were independent older adults aged ≥65 years in Japan. We used >5% weight loss during follow-up as the outcome variable and frequency of eating together as the explanatory variable. The relative risks (RRs) and 95% confidence intervals (95% CIs) were estimated based on the Poisson regression model with a Huber-White sandwich estimator for standard errors, including possible confounders., Results: among 56,919 participants, the mean age was 73.0 years (1SD = 5.5) at baseline, and 47.9% were male. About 15.1% (n = 8,596) of the participants experienced >5% weight loss during follow-up. The proportion of each category of the eating together frequency was 36.6% for 'every day', 10.3% for 'several times a week', 26.8% for 'several times a month', 20.5% for 'several times a year' and 5.8% for 'seldom'. Compared to 'every day', only 'several times a year (RR = 1.07, 95% CI = 1.01-1.13)' and 'seldom (RR = 1.17, 95% CI = 1.08-1.27)' were significantly associated with the increased risk of >5% weight loss., Conclusion: there is a temporal association between less frequent opportunities to eat together and the increased risk of weight loss among independent older adults., (© The Author(s) 2022. Published by Oxford University Press on behalf of the British Geriatrics Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Objective: To clarify the clinical significance of development of urinary abnormality in mixed connective tissue disease (MCTD)., Methods: Forty-one patients with an initial diagnosis of MCTD, followed at five hospitals between April 1, 2000 and December 31, 2013, were included. The relationship between urinary abnormality and various clinical parameters were retrospectively analyzed. Urinary abnormality was defined as proteinuria and/or hematuria detected by urinalysis. Development of other connective tissue diseases (CTDs) was defined as satisfaction of the criteria of each respective disease., Results: Of 41 patients (34 females, 7 males, mean age at diagnosis 42.2 ± 15.2 years), 16 developed urinary abnormality (UrA(+) patients). The total incidences of development of other CTDs were higher in the UrA(+) patients than UrA(-) (62.5% versus 16.0%, p = .01). In the comparison between UrA(+) and UrA(-) patients, there were no significant differences in follow-up duration or last determined estimated glomerular filtration rate (eGFR), although eGFR decreased more significantly in the UrA(+) patients than UrA(-). (-20.2 ± 17.2 vs -6.1 ± 13.8 ml/min/1.73m2, p = .01; -21.0 ± 18.9 vs -6.7 ± 14.1%, p = .03)., Conclusion: Urinary abnormality during the clinical course in MCTD is predictive of a higher incidence of developing other CTDs. Furthermore, it might also predict long-term renal prognosis in patients with an initial diagnosis of MCTD., (© 2021 Japan College of Rheumatology.)

Sidewalks are indispensable environmental resources for daily life in that they encourage physical activity. However, the proportion of sidewalk coverage is low even in developed countries. We examined the association between neighborhood sidewalk environment and dementia in Japan. We conducted a 3-year follow-up (2010-2013) among participants in the Japan Gerontological Evaluation Study, a population-based cohort study of community-dwelling older adults. We ascertained the incidence of dementia for 76,053 participants from the public long-term care insurance system. We calculated sidewalk coverage (sidewalk area as a percentage of road area) within 436 residential neighborhood units using geographic information systems. Multilevel survival models were used to estimate hazard ratios for the incidence of dementia. During follow-up, 5,310 dementia cases were found. In urban areas, compared with the lowest quartile of sidewalk coverage, the hazard ratio was 0.42 (95% confidence interval: 0.33, 0.54) for the highest quartile, adjusting for individual covariates. After successive adjustments for other neighborhood factors (land slope; numbers of hospitals, grocery stores, parks, railway stations, and bus stops; educational level; and unemployment rate), the hazard ratio remained statistically significant (hazard ratio = 0.71, 95% confidence interval: 0.54, 0.92). Living in a neighborhood with a high level of sidewalk installation was associated with low dementia incidence in urban areas., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Johns Hopkins Bloomberg School of Public Health.)

Objectives: The Childhood Health Assessment Questionnaire (CHAQ) is one of the most widely used self- report questionnaires to measure functional status in Juvenile idiopathic arthritis (JIA). The Japanese version of the CHAQ (JCHAQ) has been revised to meet requirements of clinical international trials which need the same number of questions in each functional area of the CHAQ. Methods: The original JCHAQ consisted of 36 items, measuring eight functional areas. This was changed to 30 items of questionnaire so that each functional area has same number of questions as the original US English version. The revised version was professionally translated from English to Japanese, reviewed, and validated with Japanese JIA patients. Results: A total of 42 JIA patients were enrolled in the validation: seven systemic, 30 polyarticular/oligoarticular and five enthesis related. Most patients were well controlled and the median disability index (DI) scores was 0.0 [0-0.03]; however, significant correlation was seen with visual analog scale (VAS) of pain, VAS overall well-being, physician VAS, DAS (Disease Activity Score) 28-ESR, and JADAS (Juvenile Arthritis Disease Activity Score)-27. In comparison of two groups of disease activity, remission or inactive/low disease activity vs. moderate/high disease activity, both DAS28-ESR and JADAS-27 showed significant correlation with DI. Conclusion: The updated JCHAQ was a reliable and valid tool for the functional assessment of children with JIA. It is more suitable for international and transitional comparison.

Recent studies have revealed that not only proton-sensing channels, but also one family of G protein-coupled receptors (GPCRs) comprising OGR1, GPR4, G2A and TDAG8 are responsible for the sensing of extracellular protons, or pH. Here, we report that two other GPCRs, GPR31 and GPR151, were also activated in acidic condition. Elevated pH of assay mixtures resulted in a remarkable increase in [35S]GTPγS binding by GPR31-Giα and GPR151-Giα fusion proteins in a narrow range between pH 6 and 5. Our reporter gene assays with CHO cells expressing recombinant GPR31 or GPR151 also showed that activation was maximal at pH ∼5.8. Although these results from in vitro and cellular assays revealed slightly different pH sensitivities, all of our results indicated that GPR31 and GPR151 sensed extracellular protons equally well as other proton-sensing GPCRs., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.)

The 2011 Great East Japan Earthquake and Tsunami resulted in widespread property destruction and over 250,000 displaced residents. We sought to examine whether the type of housing arrangement available to the affected victims was associated with a differential incidence of depressive symptoms. In this prospective cohort study, which comprised participants aged ≥65 years from Iwanuma as a part of the Japan Gerontological Evaluation Study, we had information about the residents' mental health both before the disaster in 2010 and 2.5 years afterward. The Geriatric Depression Scale was used. Type of accommodation after the disaster was divided into 5 categories: no move, prefabricated housing (temporary housing), existing private accommodations (temporary apartment), newly established housing, and other. Poisson regression analysis was adopted, with and without multiple imputation. Among the 2,242 participants, 16.2% reported depressive symptoms at follow-up. The adjusted rate ratio for depressive symptoms among persons moving into prefabricated housing, compared with those who did not, was 2.07 (95% confidence interval: 1.45, 2.94). Moving into existing private accommodations or other types of accommodations was not associated with depression. The relationship between living environment and long-term mental health should be considered for disaster recovery planning.

Background: Childhood socioeconomic disadvantage has been previously linked to increased mortality risk in adulthood. However, most previous studies have focused on middle-aged adults in Western contexts. Here, we sought to examine the association between childhood socioeconomic status (SES) and mortality among healthy older Japanese adults., Methods: We conducted a 3-year follow-up of participants in the Japan Gerontological Evaluation Study (JAGES), a population-based cohort of 65- to 103-year-old Japanese adults. Childhood SES was assessed by survey at baseline. Mortality from 2010 to 2013 was analysed for 15 449 respondents (7143 men and 8306 women). Cox regression models were used to estimate hazard ratios (HR) for risk of death., Results: A total of 754 deaths occurred during the 3-year follow-up. Lower childhood SES was significantly associated with lower mortality in men, but not in women. Compared with men growing up in more advantaged childhood socioeconomic circumstances, the age-adjusted HR for men from low childhood SES backgrounds was 0.75 [95% confidence interval (CI): 0.56-1.00]. The association remained significant after adjustment for height, education, adult SES, municipalities of residence, health behaviours, disease status and current social relationships (HR = 0.64; 95% CI 0.47-0.87). This association was stronger among men aged 75 years or older, HR = 0.67 (95% CI: 0.47-0.95), compared with men aged 65-74 years, HR = 0.90 (95% CI: 0.54-1.51)., Conclusions: Childhood socioeconomic disadvantage is associated with lower mortality among men aged 75 years or older, which may be due to selective survival, or alternatively to childhood physical training or postwar calorie restriction in this generation of Japanese males., (© The Author 2016. Published by Oxford University Press on behalf of the International Epidemiological Association.)

4-Methyl-5-hydroxyethylthiazole kinase (ThiM) participates in thiamin biosynthesis as the key enzyme in its salvage pathway. We purified and characterized ThiM from Escherichia coli. It has broad substrate specificity toward various nucleotides and shows a preference for dATP as a phosphate donor over ATP. It is activated by divalent cations, and responds more strongly to Co(2+) than to Mg(2+).

Background: Eating by oneself may be a risk factor for mental illness among older adults, but may be influenced by cohabitation status. We examined the association between eating alone and depression in the context of cohabitation status in older adults in Japan., Design: A longitudinal, population-based study., Setting: Data from the Japan Gerontological Evaluation Study., Subjects: We analysed 17,612 men and 19,581 women aged ≥65 without depression (Geriatric Depression Scale <5) at baseline in 2010., Methods: Eating status was classified into two categories: eating with others and eating alone. The risk of depression onset by 2013 was estimated using Poisson regression., Results: After adjusting for socioeconomic status, physical health, nutritional status, social support, social participation, frequency of meet friends, employment status and marital status, the adjusted rate ratio (ARR) for depression onset in men who ate alone compared with those who ate with others was 2.36 (95% confidence intervals [CI]: 1.18-4.71) for those living alone and 1.03 (95% CI: 0.81-1.32) for those living with others. Among women, the ARR for depression for those who ate alone compared with those who ate with others was 1.31 (95% CI: 1.00-1.72) for those living alone and 1.21 (95% CI: 1.01-1.44) for those living with others., Conclusions: Eating alone may be a risk factor for depression. Among men, the effect of eating alone on depression may be reinforced by living alone, but appears to be broadly comparable in women living alone and women living with others., (© The Author 2015. Published by Oxford University Press on behalf of the British Geriatrics Society.)

Fish have a complex self-defense mechanism against microbial invasion. Recently, l-lysine α-oxidases have been identified from a number of fish species as a novel type of antibacterial protein in the integument. These enzymes exhibit strict substrate specificity for l-lysine, but the underlying mechanisms and details of their catalytic properties remain unknown. In this study, a synthetic gene coding for Scomber japonicus l-lysine α-oxidase, originally termed AIP (for apoptosis-inducing protein), was expressed in Pichia pastoris, and the recombinant enzyme (rAIP) was purified and characterized. rAIP exhibited essentially the same substrate specificity as the native enzyme, catalyzing the oxidative deamination of l-lysine as an exclusive substrate. rAIP was N-glycosylated and remained active over a wide range of pH, with an optimal pH of 7.5. The enzyme was stable in the pH range from 4.5 to 10.0 and was thermally stable up to 60°C. A molecular modelling of rAIP and a comparative structure/sequence analysis with homologous enzymes indicate that Asp(220) and Asp(320) are the substrate-binding residues that are likely to confer exclusive substrate specificity for l-lysine on the fish enzymes., (© The Authors 2014. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.)

Glycosphingolipids (GSLs) are essential membrane components of eukaryotic cells. Recently, a new type of fungal neogala-series GSL was identified in aureobasidin A-resistant fungi. In this study, we analyzed GSLs from four pathogenic fungal strains belonging to the order Hypocreales, and found that Mariannaea elegans contained both acidic GSLs and neutral GSLs with mono- and di-saccharides. The structures of the neutral GSLs of M. elegans were determined by compositional sugar, fatty acid, and sphingoid analyses by GC/MS, MALDI time-of-flight/MS, and 1H NMR. The ceramide moiety of Glcβ1-Cer consisted mainly of the 2-hydroxylated C18:0-fatty acid 9-methyl-octadeca-4-sphinganine or 9-methyl-octadeca-4,8-sphingadienine. In contrast, the ceramides of Galβ1-6Galβ1-Cer and Glc1-6Galβ1-Cer consisted mainly of saturated 2-hydroxylated C24:0-fatty acids and C18:0-phytosphingosine. To our knowledge, Glc1-6Galβ1-Cer is a novel GSL in fungi, and M. elegans is the first example of an aureobasidin A-sensitive fungus that possesses fungal neogala series GSLs.

Background: At 1446 on 11 March 2011, northeastern Japan was struck by a major earthquake measuring 9.0 on the Richter scale. There have been several reports of transient blood pressure increases after a major earthquake, but the impact of a major earthquake on blood pressure in chronic kidney disease (CKD) patients has not been fully investigated., Methods: Changes in clinic blood pressure following the earthquake were investigated in 132 hypertensive patients with stage 3 and 4 CKD who were residents of Fukushima City., Results: Both systolic and diastolic blood pressures were significantly elevated 1-3 weeks after the earthquake compared with before the earthquake (134 ± 19 mm Hg vs. 138 ± 20 mm Hg, P = 0.02 for systolic; 76 ± 13 mm Hg vs. 79 ± 12 mm Hg, P = 0.01 for diastolic), and these values returned to baseline by 5-7 weeks after the earthquake. Multiple logistic regression analysis identified male sex (odds ratio (OR), 0.35; 95% confidence interval (CI), 0.14-0.86; P = 0.02), mean blood pressure at baseline (OR, 0.92; 95% CI, 0.86-0.96; P < 0.01), and sympatholytic medications, such as α-blockers, β-blockers, or central sympatholytics (OR, 0.23; 95% CI, 0.07-0.76; P = 0.01), as independent factors related to elevation of mean blood pressure 1-3 weeks after the earthquake in CKD patients., Conclusions: Blood pressure was significantly increased after a major earthquake in hypertensive patients with stage 3 and 4 CKD. During the first 3 weeks after the earthquake, blood pressure control was associated with the use of sympatholytic medications.

Compared to the many studies on the physiological and toxicological effects of selenium (Se) in mammals, avian Se metabolism is still an unexplored topic. Some birds are useful as poultry for human nutrition. Moreover, birds belong to higher trophic levels in the biosphere and thus may play an important role in Se circulation in the ecosystem in the same way as mammals do. In this study, we analyzed the distribution and metabolism of Se in an experimental bird, the Japanese quail, which was fed drinking water containing sodium selenite or selenomethionine (SeMet). The highest concentration of Se was detected in the pancreas, followed by down feathers, liver, and kidneys. SeMet was more efficiently incorporated into the quail than selenite. The specific and preferable distribution of Se to the high molecular weight fraction in the serum of the quail was observed only in the SeMet-ingestion group. As in mammals, selenosugar and trimethylselenonium were the major metabolites in quail excreta. Three unknown Se metabolites were detected by HPLC-ICP-MS. Although part of the metabolic pathway of Se in the Japanese quail fed selenite and SeMet was the same as that observed in mammals, the bird also showed certain avian-specific metabolic process for Se.

Background: Tissue accumulation of advanced glycation end-products (AGE) is thought to be a contributing factor to the progression of cardiovascular disease (CVD). Skin autofluorescence, a non-invasive measure of AGE accumulation using autofluorescence of the skin under ultraviolet light, has shown associations with CVD in haemodialysis patients. The present study aimed to evaluate relationships of skin autofluorescence to renal function as well as CVD in pre-dialysis patients with chronic kidney disease (CKD)., Methods: Subjects in this cross-sectional analysis comprised 304 pre-dialysis CKD patients [median age, 62.0 years; median estimated glomerular filtration rate (eGFR), 54.3 mL/min/1.73 m(2); diabetes, n = 81 (26.6%)]. AGE accumulation in skin was assessed by skin autofluorescence using an autofluorescence reader. Relationships between skin autofluorescence, eGFR, CVD history and other parameters were evaluated., Results: Skin autofluorescence correlated negatively with eGFR (r = -0.42, P < 0.01) and increased as CKD stage advanced. Multiple regression analysis revealed significant correlations of skin autofluorescence with age, presence of diabetes, eGFR and CVD history in CKD patients (R(2) = 30%). Age, male gender, smoking history, skin autofluorescence and eGFR were significantly correlated with CVD history, and multiple logistic regression analysis identified age [odds ratio (OR), 1.09; 95% confidence interval (CI), 1.03-1.15; P < 0.01], history of smoking (OR, 6.50; 95%CI, 1.94-21.83; P < 0.01) and skin autofluorescence (OR, 3.74; 95%CI, 1.54-9.24; P < 0.01) as independent factors., Conclusions: Tissue AGE accumulation measured as skin autofluorescence increased as GFR decreased and was related to CVD history in CKD patients. Non-invasive autofluorescence readers may provide potential markers for clinical risk assessment in pre-dialysis CKD patients.

Objective: Cushing's disease (CD) is usually caused by ACTH-secreting pituitary microadenomas, while silent corticotroph adenomas (SCA) are macroadenomas without Cushingoid features. However, the molecular mechanism(s) underlying their different tumor growth remains unknown. The aim of the current study was to evaluate and compare the gene expression profile of cell cycle regulators and cell growth-related transcription factors in CD, SCA, and non-functioning adenomas (NFA)., Design and Methods: Tumor tissue specimens resected from 43 pituitary tumors were studied: CD (n=10), SCA (n=11), and NFA (n=22). The absolute transcript numbers of the following genes were quantified with real-time quantitative PCR assays: CDKN2A (or p16(INK4a)), cyclin family (A1, B1, D1, and E1), E2F1, RB1, BUB1, BUBR1, ETS1, and ETS2. Protein expressions of p16 and cyclin D1 were semi-quantitatively evaluated by immunohistochemical study., Results and Conclusion: CDKN2A gene expression was about fourfold greater in CD than in SCA and NFA. The gene expressions of cyclins D1, E1, and B1, but not of A1, in CD were significantly suppressed than those in NFA. Cyclin D1 gene expression positively correlated with cyclins B1 and E1. The gene expressions of E2F1, RB1, BUB1, BUBR1, ETS1, and ETS2 did not differ between each group. Positive immunostaining for p16 and negative immunostaining for cyclin D1 were more frequent in CD than in NFA; there were positive correlations between mRNA and protein expressions of p16 and cyclin D1. Thus, it is suggested that upregulated CDKN2A with the concomitant downregulated cyclin gene family is partly involved in the small size of ACTH-secreting adenoma.

Astaxanthin possesses higher antioxidant activity than other carotenoids and, for this and other reasons, has great commercial potential for use in the aquaculture, pharmaceutical, and food industries. The basidiomycetous yeast Xanthophyllomyces dendrorhous is one of the best natural producers of astaxanthin, but wild-type cells accumulate only a small amount of astaxanthin. In this study, we developed an efficient flow cytometry method to screen for astaxanthin-overproducing mutants of X. dendrorhous. We first examined the relationship between cellular astaxanthin content and the intensity of fluorescence emitted from the cell. Although the fluorescence emission maximum of astaxanthin dissolved in acetone occurred at 570 nm, intracellular astaxanthin content correlated better with emission at around 675 nm in different X. dendrorhous strains. Using this emission wavelength, we screened cells mutagenized with ethyl methanesulfonate and successfully isolated mutants that produced 1.5-3.8-fold more astaxanthin than parent cells. This method enabled us to obtain overproducers five times more efficient than conventional screening from plate culture.

We screened soil samples for CO(2)-requiring extreme oligotrophs similar to Rhodococcus erythropolis N9T-4, which can grow on a basal salt agar medium without an organic carbon source. From 387 soil samples, three isolates were obtained and identified as Streptomyces spp. by 16S rDNA analysis. The isolates required gaseous CO(2) for growth and grew on a basal salt medium solidified by silica gel. These results suggest that such CO(2)-requiring oligotrophs occur widely in nature.

Aspergillus oryzae has two fructosyl-amino acid oxidase (FAOD) isozymes (AoFao1 and AoFao2), which are different in the substrate specificities. Northern blot analysis showed both FAO genes were induced by autoclave-browned medium containing l-lysine or l-valine. Studies with a mutant, that had a disrupted AoFAO2 gene, revealed that the expression of AoFAO1 by fructosyl l-valine depended on the expression of AoFAO2. Both genes were also induced by one of the FAOD-reaction products, glucosone. In contrast, other alpha-dicarbonyl compounds, which display a similar structure to that of glucosone were not able to induce the genes expression. These results imply that glucosone may contribute to the expression of FAO genes.

Fructosyl-amino acid oxidase (FAOD)-reactive fraction (FRY) was found in commercial yeast extract. FRY showed very hydrophilic property and was adsorbed to phenylboronate silica gel, indicating that it contained the Amadori compound. TLC and amino acid analyses revealed that glucosone, lysine, and arginine were produced from FRY after incubation with FAOD. TOF-MS analysis confirmed that FRY is a mixture of fructosyl lysine and fructosyl arginine. These compounds were also detected in mycelial extract of an FAOD-producer, Aspergillus terreus GP1, grown on the minimum medium, suggesting that a glycation reaction occurs in fungal cells and that FAOD acts toward the resultant Amadori compounds.

Zidovudine (ZDV), an antiretroviral drug used alone or in combination with other antiretroviral agents to treat HIV-infected pregnant women and their newborn infants, effectively reduces mother-to-child transmission of the virus. That myopathy and cardiomyopathy, related to mitochondrial damage, develop in some adults chronically treated with ZDV has long been known; recently, reports have suggested that similar adverse effects may occur in some infants exposed perinatally. Using a mouse model of human neonatal exposure, we treated pregnant CD-1 mice twice daily with doses of 75 mg/kg ZDV plus 37.5 mg/kg lamivudine throughout gestation and lactation; pups were exposed by direct gavage beginning postnatal day (PND) 4 and sacrificed on PND 28. Hearts were removed rapidly, and ventricles were processed for electron microscopy. Morphometric and semiquantitative morphological analyses were performed on three micrographs from each of three blocks from each of three females and three males from the control and treated groups. Treated mice showed significant increases in the mean area and decreases in the mean number of cardiomyocytic mitochondria compared to controls. We observed clusters of damaged mitochondria more frequently in treated animals than in controls; damage included fragmentation and loss of cristae. These results, demonstrating alterations in cardiomyocytic mitochondria of mice exposed in utero and postnatally, may model cardiac damage reported in human infants similarly exposed to ZDV. Critical insights derived from animal-model data like these may be used to mitigate risks to thousands of human infants receiving essential lifesaving therapy with antiretroviral drugs.

A Mn-depositing fungus, Acremonium-like hyphomycete strain KR21-2, was isolated from a Mn deposit occurring on the wall of a storage bottle containing Mn(III, IV) oxide-coated streambed pebbles and stream water. 18S rRNA gene sequence analysis revealed that strain KR21-2 was phylogenetically related to members of the order Hypocreales within the class Ascomycetes. The spent culture medium at the stationary phase of fungal growth contained a 54-kDa protein capable of depositing Mn oxides. The enzymatic activity was inhibited by azide and o-phenanthroline. The Mn(II)-oxidizing protein possessed a laccase activity, as indicated by direct oxidation of p-phenylenediamine and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid). These results are consistent with the role assumed for laccase-like multicopper oxidase, which is proposed to be involved in the Mn(II)-oxidizing factors from some bacteria. Unlike laccases of basidiomycete fungi, however, the protein of strain KR21-2 did not produce soluble Mn(III) species in the presence of either of the Mn chelators pyrophosphate and malonate. This is the first report on the possible involvement of laccase and/or multicopper oxidase in Mn oxide deposition by ascomycetes (including their anamorphs) ubiquitous in natural environments.

Cord blood (CB) cells are a useful source of hematopoietic cells for transplantation. The hematopoietic activities of CB cells are different from those of bone marrow and peripheral blood (PB) cells. Platelet recovery is significantly slower after transplantation with CB cells than with cells from other sources. However, the cellular mechanisms underlying these differences have not been elucidated. We compared the surface marker expression profiles of PB and CB hematopoietic cells. We focused on two surface markers of hematopoietic cell immaturity, i.e., CD34 and AC133. In addition to differences in surface marker expression, the PB and CB cells showed nonidentical differentiation pathways from AC133(+)CD34(+) (immature) hematopoietic cells to terminally differentiated cells. The majority of the AC133(+)CD34(+) PB cells initially lost AC133 expression and eventually became AC133(-)CD34(-) cells. In contrast, the AC133(+)CD34(+) CB cells did not go through the intermediate AC133(-)CD34(+) stage and lost both markers simultaneously. Meanwhile, the vast majority of megakaryocyte progenitors were of the AC133(-)CD34(+) phenotype. We conclude that the delayed recovery of platelets after CB transplantation is due to both subpopulation distribution and the process of differentiation from AC133(+)CD34(+) cells.

Abstract We describe the case of a Japanese girl who suffered from both Behçet's disease (BD) and bilateral sacroiliitis. This patient was diagnosed with BD at 8 years of age, and began to complain of buttock pain when she was 10 years old. X-rays of her pelvis showed bilateral grade 3 sacroiliitis, and bone scintigraphy showed an increased uptake in the sacroiliac joints. Movements of the spine and chest expansion were not restricted. HLA typing was positive for B44 and DR4, but negative for B27. The patient was not diagnosed as having seronegative spondyloarthropathy. This case indicates that erosive sacroiliitis is an intrinsic condition of BD.

We describe an application of gel microdroplet (GMD) and flow cytometry techniques to selective enrichment of non-growing Leuconostoc mesenteroides cells, which are well culturable on other media, from a mixture with Bacillus subtilis cells in nutrient broth. After encapsulating cells of the mixed population within GMDs and a brief incubation in nutrient broth, the inability of L. mesenteroides cells to form microcolonies within GMDs allowed their discrimination from B. subtilis cells. After staining the GMD mixture with 6-carboxyfluorescein diacetate, which showed no influence on cell viability, the GMDs containing single cells of L. mesenteroides were selectively collected using flow cytometry sorting based on differences in fluorescence intensity. The cells of L. mesenteroides retained viability during the process.

AC133 antigen is a novel marker for human hematopoietic stem/progenitor cells. In this study, we examined the expression and proliferation potential of AC133(+) cells obtained from steady-state peripheral blood (PB). The proportion of AC133(+) cells in the CD34(+) subpopulation of steady-state PB was significantly lower than that of cord blood (CB), although that of cytokine-mobilized PB was higher than that of CB. The proliferation potential of AC133(+)CD34(+) and AC133(-)CD34(+) cells was examined by colony-forming analysis and analysis of long-term culture-initiating cells (LTC-IC). Although the total number of colony-forming cells was essentially the same in the AC133(+)CD34(+) fraction as in the AC133(-)CD34(+) fraction, the proportion of LTC-IC was much higher in the AC133(+)CD34(+) fraction. Virtually no LTC-IC were detected in the AC133(-)CD34(+) fraction. In addition, the features of the colonies grown from these two fractions were quite different. Approximately 70% of the colonies derived from the AC133(+)CD34(+) fraction were granulocyte-macrophage colonies, whereas more than 90% of the colonies derived from the AC133(-)CD34(+) fraction were erythroid colonies. Furthermore, an ex vivo expansion study observed expansion of colony-forming cells only in the AC133(+)CD34(+) population, and not in the AC133(-)CD34(+) population. These findings suggest that to isolate primitive hematopoietic cells from steady-state PB, selection by AC133 expression is better than selection by CD34 expression.

A water-soluble ester, methyl formate, was detected as a metabolite in the culture medium of methylotrophic yeasts. Methyl formate synthase, which catalyses NAD(+)-dependent dehydrogenation of the hemiacetal adduct of methanol and formaldehyde, catalyses the ester synthesis. The enzyme activity was induced on a methanol medium and was increased further by the addition of formaldehyde. In the reaction system using intact cells of Pichia methanolica AKU 4262, 135 mM (8.1 g/liter) methyl formate was produced from 2 M methanol. This is a new biological process for ester synthesis that couples spontaneous formation of hemiacetal and alcohol dehydrogenase.

We investigated IgG, IgA and IgM class specific antibodies to five bacterial (Klebsiella pneumoniae, Escherichia coli, Salmonella enteritidis, Salmonella typhimurium and Shigella flexneri) lipopolysaccharides (LPS) by enzyme-linked immunosorbent assay in 144 Japanese patients with ankylosing spondylitis (AS). AS patients had significantly elevated IgA antibodies to K. pneumoniae LPS, Salmonella enteritidis LPS and Salmonella typhimurium LPS; however, there was no correlation between antibody level to LPS and acute-phase reactants, erythrocyte sedimentation rate and serum C-reactive protein.

T4-binding globulin (TBG) is the major transport protein of thyroid hormone in man. Inherited TBG abnormalities were manifested fully in hemizygous males and partially in heterozygous females and transmitted in an X-chromosome-linked fashion, compatible with its location on Xq21-22. We have previously reported that complete deficiency (CD) and partial deficiency (PD) in Japanese subjects resulted from two distinct mutations of the TBG gene, TBG-CDJ and TBG-PDJ, respectively. Recently, we encountered a female manifesting TBG-CD and herein investigated the molecular mechanisms. She was found to possess TBG-CDJ and common-type TBG (TBG-C) alleles by characterizing the TBG gene. Then, X-chromosome inactivation status was evaluated in her family members using a phosphoglycerate kinase (PGK) gene, located on Xq13. Three TBG-CDJ heterozygotes and one unaffected female, confirmed to be PGK heterozygotes for a polymorphic BstXI site, were analyzed. Only the CD female was shown to undergo selective inactivation by examining the BstXI site in amplified products after digestion with a methylation-sensitive enzyme, HpaII. Among an additional eight informative females with TBG deficiency, one heterozygous female for TBG-PDJ shared this selective inactivation pattern. Moreover, the X-chromosome with TBG-C was suggested to be inactivated selectively from the linkage of PGK and TBG alleles recognized in eight of nine family members. Selective X-chromosome inactivation was considered to be the cause of a female heterozygous for TBG-CDJ or -PDJ manifesting the same phenotype as a hemizygote.

In Fusarium oxysporum S-1F4, fructosyl lysine oxidase (FLOD) was induced with N epsilon-fructosyl N alpha-Z-lysine (epsilon-FL), which is a model compound of a glycated protein, and the induction was inhibited by the addition of cycloheximide in the growing cells. FLOD formation was greatly enhanced in an autoclave-browned medium containing glucose and L-lysine. Some Amadori compounds formed from glucose and L-lysine during autoclaving were assumed to induce the enzyme. After optimization of the culture conditions, FLOD produced in the browned medium was comparable to that in the medium with epsilon-FL.

A considerable amount of methylformate accumulated in the culture medium of methanol-grown methylotrophic yeasts. Methylformate is considered as an intermediate in a novel formaldehyde oxidation pathway. Through investigations with Pichia methanolica, methylformate formation was found to be catalysed by a new type of alcohol dehydrogenase, which was named methylformate synthase. When cells were grown on a relatively high concentration of methanol or exposed to a high concentration of formaldehyde, formation of methylformate was enhanced and the level of methylformate synthase in the cells increased. How methylformate synthase is involved in formaldehyde oxidation and formaldehyde detoxification is discussed.

Fructosyl lysine oxidase (FLOD) was examined for its use in the enzymatic measurement of the level of glycated albumin in blood serum. To isolate microorganisms having such an enzyme activity, we used N epsilon-fructosyl N alpha-Z-lysine (epsilon-FL) as a sole nitrogen source in the enrichment culture medium. The isolated fungus, strain S-1F4, showed a high FLOD activity in the cell-free extract and was identified as Fusarium oxysporum. FLOD was purified to an apparent homogeneity on SDS-PAGE. The molecular mass of the subunit was 50 kDa on SDS-PAGE and seemed to exist in a monomeric form. The enzyme had an absorption spectrum characteristic of a flavoprotein and the flavin was found to be covalently bound to the enzyme. The enzyme acted against N epsilon-fructosyl N alpha-Z-lysine and N alpha-fructosyl N epsilon-Z-lysine and showed specificity for fructosyl lysine residues.

Saccharomyces cerevisiae wy2 segregated to 2 mater and 2 non-mater in relation to mating ability. The non-mater segregants behaved as the normal type of homothallic life cycle. On the other hand, the mater segregants gradually formed spores during successive subcultures, indicating that slow interconversion of mating-type happened to occur during subcultures. We termed this novel type of life cycle "delayed homothallism". The results of complementation tests with standard ho strains and introduction of a wild type HO gene showed that delayed homothallism was caused by a defective HO gene. The amino acid sequence deduced from the nucleotide sequence of the wy2 HO gene differed from the wild type HO gene in three amino acid residues. In the carboxy terminus of HO protein, there are three repeats of cysteine and histidine that are postulated to play a role in binding of HO protein to DNA. However, wy2 HO protein lacked one such repeat at residues Cys470-His475, where His was replaced by Leu.

We cloned the rat T4-binding globulin (rTBG) gene, characterized its organization, and studied its promoter activity and regulation. A genomic DNA library was constructed and screened using an rTBG complementary DNA (cDNA) as a probe. An 8.6-kilobase pair (kbp) clone was partially sequenced and compared with the sequence of the previously cloned cDNA. It helped complete the cDNA sequence and identify the first noncoding exon (exon 0). The transcription start site was identified using an RNase protection assay. The rTBG genomic clone contained 1.2 kbp 5'-flanking and 1.7 kbp 3'-flanking regions. The sizes of exons and introns of the rTBG gene are similar to those of the human TBG gene, belonging to the serine protease inhibitor family. The 5'-flanking region contains a TATA box, a CAAT box, and a consensus sequence for the hepatocyte nuclear factor 1-binding site. We tested the promoter activity of the 1.2-kbp 5'-flanking region using a luciferase reporter plasmid. When transfected into a hepatocyte-derived cell line (HepG2), the plasmid construct containing the fragment -1227 to +11 (transcription start site, +1) showed a 9-fold increase in luciferase activity compared with that of a promoterless luciferase vector. No promoter activity was detected in a nonhepatocyte-derived cell line (COS1). Serial 5'-deletion revealed that the construct containing the fragment -180 to +11 had 40% of the maximal promoter-induced luciferase activity. And that containing the fragment -53 to +11 showed no significant increase of luciferase activity. These results suggest that the region -180 to -53, containing hepatocyte nuclear factor 1-binding site, is essential to the liver-specific expression. We previously reported down-regulation of rTBG messenger RNA by T3 in vivo. The present study failed to show T3's effect on the promoter activity of the 1.2-kbp 5'-flanking region of the rTBG gene.

We have previously reported a Japanese family manifesting partial TBG deficiency (TBG-PDJ). This variant was characterized by a decreased level of serum TBG concentration, heat lability, and normal isoelectric focussing pattern, but the affinity to iodothyronine is unknown. The TBG-PDJ gene possesses a single nucleotide substitution replacing the normal Pro363(CCT) with Leu(CTT); however, the precise mechanism that results in the reduction of the TBG concentration in the serum of the subjects harboring this mutation remains unknown. This was investigated in the current communication by expressing the complementary DNAs of TBG-PDJ and the common type TBG (TBG-C) in COS-1 cells. Pulse-chase experiments revealed impaired secretion of TBG-PDJ. TBG-C secretion into the medium was evident during 60 min of the pulse period and was almost completed by 12 h. On the other hand, TBG-PDJ was secreted slowly and continued to accumulate between 12-24 h of the chase period. The molecular mass of TBG-PDJ in the cell lysate was identical to that of TBG-C when estimated by gel electrophoresis (54 kilodaltons). The content of TBG-PDJ in the cell lysate decreased less rapidly than that of TBG-C, indicating that impaired TBG-PDJ secretion accounts for the partial TBG deficiency. Oligosaccharide units of intracellular TBG-C were resistant to endoglycosidase-H, but half of those of TBG-PDJ were sensitive to the enzyme digestion, suggesting partial retention of TBG-PDJ within the rough endoplasmic reticulum. Northern blot analysis revealed abundant messenger ribonucleic acid for the glucose-regulated protein-78, the level of which was 3.54-fold greater in the cells transfected with TBG-PDJ than in nontransfected COS-1 cells, whereas that in TBG-C-transfected cells was same as that in the nontransfected cells. Increased expression of glucose-regulated protein-78 together with the sensitivity to endoglycosidase-H suggests impairment of intracellular processing of TBG-PDJ. Our results indicate that the impaired intracellular transport of the TBG-PDJ molecule is the main cause of the reduced concentration of immunoreactive TBG in the serum of subjects harboring this TBG variant.

We have previously reported six unrelated Japanese families having the same mutation in the TBG gene and manifesting complete TBG deficiency (TBG-CDJ). The deficiency consists of a single nucleotide deletion resulting in the production of C-terminal truncation due to a frameshift and premature termination. However, the reason for the failure to detect TBG in the serum of subjects harboring this mutation remains unknown. In this communication we investigated the mechanism of the TBG deficiency associated with TBG-CDJ. The complementary DNAs of TBG-CDJ and normal TBG (TBG-N) were expressed in COS-1 cells. Pulse-chase experiments revealed a complete absence of secretion of TBG-CDJ, whereas TBG-N was already present in the medium at time 0 and was almost entirely secreted by 3 h. In cell lysates, TBG-CDJ had a smaller molecular mass [52 kilodalton (kDa)] than TBG-N (54 kDa) and gradually decreased during the chase. Thus, failure of TBG-CDJ secretion accounts for the complete TBG deficiency. The molecular mass of TBG-CDJ (42 kDa) was also smaller than that of TBG-N (44 kDa) when synthesized in the presence of tunicamycin. These findings are consistent with the premature termination of the TBG-CDJ molecule deduced from the nucleotide sequence analysis. Intracellular TBG-N was resistant to endoglycosidase H but not TBG-CDJ, suggesting the retention of TBG-CDJ within the rough endoplasmic reticulum. Indeed, subcellular fractionation revealed that most of TBG-CDJ was located in the rough endoplasmic reticulum compartment, and TBG-N was distributed in the Golgi fractions. Our results suggested that the lack of intracellular transport of the truncated TBG molecule is the cause for the absence of immunoreactive TBG in the serum of subjects harboring the TBG-CDJ variant.

A mutant strain of a methylotrophic yeast, Candida boidinii UV-16, which was obtained and characterized as a respiration-deficient mutant, showed an improved productivity of glycerol from methanol. Cultural conditions for glycerol production was studied. Feedings of methanol and a nitrogen source during cultivation promoted the glycerol production by strain UV-16. Biotin and thiamin exclusion from the culture medium repressed the growth but doubled the glycerol production. The presence of slight amounts of the vitamins from seed culture was necessary and stimulative for the glycerol production. Addition of antifoam stimulated the ascent of glycerol productivity. Under the best culture conditions, strain UV-16 produced 2.18g/liters of glycerol in a methanol medium.

The clinical effectiveness of bioclean rooms was studied in relation to prevention of infection during treatment for induction of remission of acute leukemia. According to the steps in clean patient care, patients were divided into three groups, Group A consisting of 106 patients who were accommodated in laminar air flow (LAF) rooms under strictly clean nursing, Group B consisting of 99 patients who were treated in disinfected rooms with LAF without any clean nursing techniques and Group C consisting of 188 patients who stayed in conventional wards. Since all patients were treated with antileukemic drugs and preventive administration of antimicrobial drugs under the same regimen, any difference between groups should be attributable to LAF or LAF and clean nursing. A marked and stepwise reduction in the incidence of infection between Groups A, B and C was observed. A statistically significant reduction in pneumonia, upper respiratory tract infection, periproctitis and skin abscesses was also observed in Groups A and B. As to the causative organisms, a relative increase in incidence of infections caused by Pseudomonas aeruginosa and Escherichia coli and a decrease in those caused by Proteus and fungi were observed in Groups A and B, as compared with Group C. These data suggest that pneumonia etc. may be exogenous infections and that Proteus and fungi are exogenous organisms, while Pseudomonas aeruginosa and Escherichia coli are endogenous bacteria. These data seem very helpful for setting up several steps in bioclean rooms for effective and economic patient care.

Human leukemic cells which bear antigens present on the common acute lymphocytic leukemia (ALL) cell surface can be lysed with murine monoclonal antibodies and baby rabbit complement. With NALM-6 cells used as target cells, the optimal conditions for completely eliminating the cells in vitro have been defined. The antibody cocktail plus complement is more effective than any single antibody plus complement in lysing the target cells. Optimal lysis is obtained when the antibody cocktail at 10 micrograms/ml for each antibody and baby rabbit complement at a final dilution of 1:6 are used. Leukemic cells from two newly diagnosed ALL patients were also effectively killed by this treatment. Of special interest is the finding that human complement can be used in place of baby rabbit complement.

Thirty-two adult patients with minimal change nephrotic syndrome (MCNS) were studied in order to clarify the characteristics of the immune system in MCNS and their relation to clinical activity. In the active phase (n = 17), serum immunoglobulin (Ig) M and E levels, B lymphocytes (surface Ig-positive cells) and their subsets, surface IgG, IgM and IgE positive cells; B gamma, B mu and B epsilon, were increased, whereas the serum IgG level and OKT3-reactive cells, peripheral T lymphocytes; T3, were decreased. In the remitted phase maintained by steroid therapy (n = 17), serum Igs and B lymphocyte subsets tended to return to normal levels concomitant with decreases in T3 and T4, and an increase in T8 in consequence of a marked decrease in T4/T8 (helper/suppressor) ratio. In stable remission continuing with no steroid therapy (n = 14), the above abnormalities returned to normal ranges, except for serum IgM which remained at a high level and re-elevated serum IgE. These results suggest that immunological abnormalities in MCNS are characterized by acceleration of the IgE and IgM producing systems and impaired maturation of the IgG producing system despite normal differentiation from the IgM producing to IgG producing system, possibly caused by T lymphocyte dysfunction.

Thermodynamic quantities of the self-association of 6-methylpurine in water (1)-dimethylsulfoxide (DMSO) (2) mole fraction, x2 less than 0.1) and water (1) - N,N-dimethylformamide (DMF) (2) (x2 less than or equal to 1.0) mixed solvents have been obtained through heat of dilution measurements, at 25 degrees C. In the water-DMSO solvent system, the standard enthalpy and entropy changes, delta Ho and delta So, of the association exhibited an abrupt behavior. They decreased remarkbly with the the mole fraction of DMSO until about x2 = 0.012 and after that, they increased steeply. In the case of water-DMF solvent system, the values of delta Ho and delta So didn't show the abrupt behavior. They decreased steeply until about x2 = 0.1 and, at higher mole fractions, became relatively constant. These behaviors are rationalized on the basis of solvent structural effects and solvation in these association systems.

The proportion of B lymphocyte subsets with surface immunoglobulin G (sIgG) was significantly increased in minimal change nephrotic syndrome (MCNS), membranous nephropathy, IgA nephropathy and mesangiocapillary glomerulonephritis (MCGN) and with sIgA in IgA nephropathy and MCGN, and with sIgE in MCNS. Increased subsets in membranous nephropathy, IgA nephropathy and MCGN corresponded to the immunoglobulins deposited in the glomeruli, and the increased subset of sIgE in MCNS was correlated with the elevation of serum IgE. These results suggest that each disease studied has a characteristic subset pattern of B lymphocyte response. This may have an important role in determining the histological type of idiopathic glomerulonephritis.