Non-invasive transcriptomic analysis using mRNAs in skin surface lipids obtained from children with mild-to-moderate atopic dermatitis.

BACKGROUND: Specimens for analysing the molecular pathology of skin disease are generally obtained through invasive methods, such as biopsy. However, less burdensome methods are desirable for paediatric patients. We recently established a method that comprehensively analyses RNA present in sebum (skin surface lipid-RNAs: SSL-RNAs) using a next-generation sequencer. Using this method, biological information can be obtained from the skin in a completely non-invasive manner. OBJECTIVES: To verify the applicability of the SSL-RNA method for analysis of paediatric skin and analyse the molecular pathology of mild-to-moderate atopic dermatitis (AD) in children. METHODS: We collected sebum specimens from the whole faces of 23 healthy children and 16 children with mild-to-moderate AD (eczema area and severity index (EASI) score: 5.9 ± 2.6) ranging in age from 6 months to 5 years, using an oil-blotting film. We then extracted SSL-RNAs from the samples and performed an AmpliSeq transcriptomic analysis. RESULTS: The expressions of genes related to keratinization (LCE, PSORS1C2, IVL and KRT17), triglyceride synthesis and storage (PLIN2, DGAT2 and CIDEA), wax synthesis (FAR2), ceramide synthesis (GBA2, SMPD3 and SPTLC3), antimicrobial peptides (DEFB1) and intercellular adhesion (CDSN), all of which are related to the skin barrier, are lower in children with AD than in healthy children. The children with AD also have higher expression of CCL17, a Th2-cytokine and an increased Th2-immune response as demonstrated by a gene set variation analysis. Moreover, KRT17 and CCL17 expression levels are significantly correlated with the EASI score. CONCLUSIONS: Molecular changes associated with abnormal immune responses and the epidermal barrier in children with mild-to-moderate AD can be determined using the SSL-RNA method. This non-invasive method could therefore be a useful means for understanding the molecular pathology of paediatric AD.

Evaluation of effective energy using radiochromic film and a step-shaped aluminum filter.

Although the half-value layer (HVL) is one of the important parameters for quality assurance (QA) and quality control (QC), constant monitoring has not been performed because measurements using an ionization chamber (IC) are time-consuming and complicated. To solve these problems, a method using radiochromic film and step-shaped aluminum (Al) filters has been developed. To this end, GAFCHROMIC EBT2 dosimetry film (GAF-EBT2), which shows only slight energy dependency errors in comparison with GAFCHROMIC XR TYPE-R (GAF-R) and other radiochromic films, has been used. The measurement X-ray tube voltages were 120, 100, and 80 kV. GAF-EBT2 was scanned using a flat-bed scanner before and after exposure. To remove the non-uniformity error caused by image acquisition of the flat-bed scanner, the scanning image of the GAF-EBT2 before exposure was subtracted after exposure. HVL was evaluated using the density attenuation ratio. The effective energies obtained using HVLs of GAF-EBT2, GAF-R, and an IC dosimeter were compared. Effective energies with X-ray tube voltages of 120, 100, and 80 kV using GAF-EBT2 were 40.6, 36.0, and 32.9 keV, respectively. The difference ratios of the effective energies using GAF-EBT2 and the IC were 5.0%, 0.9%, and 2.7%, respectively. GAF-EBT2 and GAF-R proved to be capable of measuring effective energy with comparable precision. However, in HVL measurements of devices operating in the high-energy range (X-ray CT, radiotherapy machines, and so on), GAF-EBT2 was found to offer higher measurement precision than GAF-R, because it shows only a slight energy dependency.

Evaluation of effective energy for QA and QC: measurement of half-value layer using radiochromic film density.

The effective energy of diagnostic X-rays is important for quality assurance (QA) and quality control (QC). However, the half-value layer (HVL), which is necessary to evaluate the effective energy, is not ubiquitously monitored because ionization-chamber dosimetry is time-consuming and complicated. To verify the applicability of GAFCHROMIC XR type R (GAF-R) film for HVL measurement as an alternative to monitoring with an ionization chamber, a single-strip method for measuring the HVL has been evaluated. Calibration curves of absorbed dose versus film density were generated using this single-strip method with GAF-R film, and the coefficient of determination (r2) of the straight-line approximation was evaluated. The HVLs (effective energies) estimated using the GAF-R film and an ionization chamber were compared. The coefficient of determination (r2) of the straight-line approximation obtained with the GAF-R film was more than 0.99. The effective energies (HVLs) evaluated using the GAF-R film and the ionization chamber were 43.25 keV (5.10 mm) and 39.86 keV (4.45 mm), respectively. The difference in the effective energies determined by the two methods was thus 8.5%. These results suggest that GAF-R might be used to evaluate the effective energy from the film-density growth without the need for ionization-chamber measurements.

Half-value layer measurement: simple process method using radiochromic film.

Although the half-value layer (HVL) is one of the important parameters for QA and QC, constant monitoring has not been performed because the measurements using an ionization chamber (IC) are time-consuming and complicated. To solve these problems, the use of radiochromic film (GAFCHROMIC XR TYPE R: GAF-R) with step-shaped aluminum (Al) filters, referred to herein as the simple process method, has been developed. The measurement X-ray tube voltages were 120 kV, 100 kV, and 80 kV. The Al filter area, the full exposure area, and the unexposed area were set on the GAF-R so as to obtain correct data. The HVL was evaluated using the density attenuation ratio. The HVLs obtained using the GAF-R and an 1C dosimeter were compared. HVLs with X-ray tube voltages of 120 kV, 100 kV, and 80 kV using the GAF-R were 4.10 mm, 3.55 mm and 2.97 mm, respectively. The difference ratios of the HVLs using the GAF-R and the IC were 1.2%, 7.6%, and 10.0%, respectively. The HVL at 120 kV can be routinely and quickly measured using the simple process method. Therefore, an IC dosimeter is not needed for HVL measurements for QA and QC. However, the HVL measurements of low energy (100 kV and 80 kV) need attention.

Antimyeloma effects of a novel synthetic retinoid Am80 (Tamibarotene) through inhibition of angiogenesis.

In multiple myeloma (MM), the interaction between myeloma cells and bone marrow microenvironment has an important role in the pathogenesis of MM. We first examined the inducing effect of myeloma cells on migration of human umbilical vein vascular endothelial cells (HUVECs). Five myeloma cell lines produced varying amounts of VEGF, and migration of HUVECs was induced by coculture with myeloma cells. We next examined the inhibitory effect of a novel synthetic retinoid Am80 (Tamibarotene) on both myeloma cells and HUVECs. Am80 is specific for the retinoic-acid receptor-alpha/beta, and has therapeutic effects in all-trans retinoic acid resistant acute promyelocytic leukemia. Am80 slightly inhibited the growth of both myeloma cells and HUVECs, and remarkably inhibited the growth of HUVECs stimulated by VEGF. Am80 showed little growth inhibition of bone marrow stromal cells (BMSCs), but it markedly inhibited migration of HUVECs by cocultured myeloma cells. Am80 inhibited VEGF-induced phosphorylation of VEGF receptor. In addition, VEGF-induced formation of tube-like structures in vitro and neovascularization in mouse corneas were significantly inhibited by Am80. These findings clearly demonstrate that Am80 is a potential inhibitor of angiogenesis caused by the interaction between vascular endothelial cells and myeloma cells, and might be a useful therapeutic agent against MM.

Studies on methylthymol blue-I Separation and purification of methylthymol blue and semimethylthymol blue.

The metallochromic indicator Methylthymol Blue was purified chromatographically with cellulose and ion-exchange resin columns. The monosubstituted product of the reaction, Semimethylthymol Blue, was also separated and purified, and can also be used for the colorimetric determination of metals. There are not marked differences between the infrared spectra of Methylthymol Blue and Semimethylthymol Blue. The purities of Methylthymol Blue and Semimethylthymol Blue finally obtained were above 97% and 90% respectively. In acidic medium Methylthymol Blue forms 1:1 greyish blue and 1:2 pure blue chelates with copper(II) whilst Semimethylthymol Blue forms only 1:1 orange chelates with bivalent metals. The molar absorptivities at 435 mmu are 1.89 (+/- 0.03) x 10(4) for Methylthymol Blue at pH 5.00 and 1.76 (+/- 0.03) x 10(4) for Semimethylthymol Blue at pH 5.45.

[An evaluation of the biting force, the body composition and the amount of masticatory action in young females].

UNLABELLED: The purposes of this study were to investigate factors related to maximum biting force and to understand the characteristics of physical properties of daily ingested foods in young females. One hundred and forty subjects aged 18-23, with Angle 1 class occlusion, had not suffered from periodontitis, and had not been treated for preparation of tooth crown of first molars. Body height and weight were measured, and percentage of body fat, fat mass (FM) and lean body mass (LBM) were estimated, using the impedance analyzer. The maximum biting force was measured by the press sensation method. According to the formula on the basis of our new version of Yanagisawa's food classification, the mean value of the amount of masticatory action for one day was calculated. Subjects were divided into the normal biting force and the low biting force groups with -1SD of the maximum biting force, in order to compare body composition and backgrounds in sports activities between these two groups. Multiple linear regression analysis was carried out, employing maximum biting force (kg.f) as a dependent variable, and having a background in sports activities, FM, LBM, the number of missing teeth, the number of dental caries and the amount of masticatory action for one day as independent variables. Results were as in the following: 1) The proportion of subjects who had a background in sports activities in the low biting force group were less than that in the normal biting force group (p < 0.01). 2) Having a background in sports activities and LBM were positively correlated to maximum biting force (p < 0.01), while the amount of masticatory action for one day was not. 3) All subjects, especially those in the low biting force group seldom had food requiring the highest amount of masticatory action. CONCLUSION: Having a background in sports activities and LBM are positively correlated with the maximum biting force, while the amount of masticatory action for one day was not correlated positively nor negatively in young females.

[Case-control study of risk factors for cedar pollinosis among parents of 3-year-old children].

PURPOSE: To identify risk factors associated with cedar pollinosis among parents of three-year-old children. METHODS: The subjects were parents whose children underwent health examination at the age of three years in September and October 1997 in Tochigi prefecture. We distributed questionnaires to the examinees beforehand with the request that they be filled in and brought to the examination site. Information on parents' environmental factors (place of residence, type and structure of housing, and distance from roads with heavy traffic), and mothers' past history and family history of allergic diseases was obtained. We defined cedar pollinosis in terms of three symptoms, sneezing, nasal discharge, and nasal obstruction, between February and May. Controls were those who did not have any of the three symptoms. Environmental factors were assessed with univariate and multivariate analyses using unconditional logistic models. Maternal genetic factors were also analyzed with odds ratios and 95% confidence intervals calculated for each. We also compared odds ratios of environmental factors between groups with and without genetic factors. RESULTS: Of the parents of children taking the health examination, 90.2% took part (2,968 of 3,291 couples). Information was obtained from 2,846 mothers and 2,905 fathers. Mother and father cases were 312 and 229, and controls were 1,857 and 1,934, respectively. For the univariate analysis of environmental factors, place of residence (residential area/agricultural area), type of housing (apartment complex/solitary house), structure (reinforced concrete building/wooden house), and distance from heavy traffic (< 100 m/100 m+) were positively related to cedar pollinosis. For the multivariate analysis using unconditional logistic models, the odds ratio was significantly high for distance from heavy traffic (< 100 m/100 m+) among fathers. Mothers' past history and family history of allergic diseases showed high odds ratios for cedar pollinosis. We classified groups with and without past and family histories of allergic diseases (group with and without genetic factors) for assessment with multivariate analyses. Odds ratios for the group with an allergic history were higher than for those without such a history, but difference for factors such as, distance from heavy traffic type of housing, and structure were not statistically significant. CONCLUSION: Risk of cedar pollinosis increases with distance from heavy traffic among fathers. Mothers with histories of allergy show slightly elevated odds ratios for environmental factors, but without statistic significance.

[Relationship between breast milk and atopic dermatitis in children].

OBJECTIVE: To determine whether or not dioxins and furans in breast milk have a role in the prevalence of atopic dermatitis among children. METHODS: The target population of the study was all children participating in health check-up program for 3-year-old children in Tochigi Prefecture in September and October 1997. Using a questionnaire, information on nutrition in infants (breast milk only, bottled milk only, or mixed), parity, mothers' age at birth, and a history of atopic dermatitis was obtained. Besides, data on potential confounding factors were obtained. RESULTS: Questionnaires from 2,968 children (85.3% of those who were to participate in the programs, and 90.2% of children who participated them) were analyzed. The risk of atopic dermatitis was higher among children with breast milk (odds ratio [OR] = 1.37 with 95% confidence interval [CI] 1.02-1.83) and those with mixed nutrition (OR = 1.21, 95% CI: 0.94-1.57) in comparison with children with only bottled milk. Mothers' age at birth (OR for those who were more than 30 years or older in comparison with those who were younger than 30 years = 1.27; 95% CI, 1.01-1.62) and those with second or later parity orders (OR = 1.32, 95% CI; 1.04-1.67) were also risk factors of the dermatitis after the adjustment for some potential confounding factors. CONCLUSION: Breast milk elevates the risk of atopic dermatitis slightly; the risk is, however, higher in children in second or later parity orders. If the PCDDs and PCDFs in breast milk cause the dermatitis, this would contradict the assumed metabolism of these chemicals in human bodies.

Reducing motion artifacts during hepatic DSA.

In hepatic digital subtraction angiography, artifacts caused by diaphragmatic motion can interfere with the correct diagnosis of hepatic lesions. This article describes the feasibility of using the exhalation holding method in reducing diaphragmatic motion artifacts in hepatic DSA. The authors compared respiratory holding methods in 30 patients who underwent hepatic DSA. They performed a blind comparison of images to assess diaphragmatic motion artifacts. Results showed the images obtained by the exhalation holding method were superior to those obtained by the inhalation holding method in 20 out of 30 series (66.7%).

Effect of ATP-binding cassette subfamily B member 1 on bovine blastocyst implantation.

The ATP-binding cassette subfamily B member 1 (ABCB1) is an efflux transporter that excretes xenobiotics and waste matter. High expression of ABCB1 induced by forskolin (FSK) and rifampicin (RIF) in the bovine blastocysts reportedly improves the cellular quality. In the present study, interferon-α, similar to FSK and RIF, was highly potent in inducing the expression of ABCB1 in the bovine blastocysts but did not exhibit an additive effect with FSK and RIF. Bovine blastocysts stimulated by the combined treatment with FSK, RIF, and interferon-α to express high levels of ABCB1 displayed better freezing resistance as indicated by higher cell numbers in post thawing cultures. On transfer to recipients, such embryos established pregnancies with significantly higher frequencies in repeat breeder cows rather than normal ones.

Congenital adrenal hyperplasia with 47, XXY Klinefelter syndrome.

We describe here a unique case of congenital adrenal hyperplasia (CAH) accompanied by Klinefelter syndrome. A Japanese boy was diagnosed as having CAH caused by 21-hydroxylase deficiency at birth, but was untreated thereafter until age 10. In the meantime he showed marked acceleration in somatic growth with sexual precocity by age 9, at which time growth completely stopped. During regular follow-ups at our clinic and steroid treatment after age 10, he was recognized as having bilateral small and firm testes. A chromosomal examination and a testicular biopsy revealed a complication of 47, XXY Klinefelter syndrome with CAH. Association of these two diseases has not been reported so far, and the present case is the first one to our knowledge.

Effects of exercise for 1 month on serum lipids in adolescent females.

BACKGROUND: The present study was done to clarify the effects of 1 month of exercise on levels of total cholesterol (TC) and high-density lipoprotein cholesterol (HDLC) and on the ratio TC/HDLC and also to evaluate the relationship of body fat to amount of exercise and TC/HDLC ratio. METHODS: Twenty-seven female athletes (aged 15-18 years) were divided into two groups: the participant's group, in which players trained and attended a tournament, and the non-participant's group, in which players did not attend the tournament. We assessed the amount of exercise, body composition, serum TC and HDLC and TC/HDLC on four occasions: before (T0), during (T1), 1 day after (T2) and a week after (T3) the experimental period. Levels of TC and HDLC adjusted for changes in plasma volume were compared for each occasion. Two multiple regression models for change in TC/HDLC from T0 to T2 and from T0 to T3 were employed. RESULTS: (i) The changing patterns in TC and HDLC throughout the program were different between the two groups; (ii) the decreased level of TC/HDLC after 1 month of exercise may easily revert to its original level; and (iii) the relevant factor for the decline in TC/HDLC was the amount of exercise, not body fat reduction. CONCLUSIONS: Exercise may be a more important factor for the improvement of TC/HDLC than concomitant body fat reduction and non-strenuous exercise may maintain a more stable and higher HDLC level than strenuous exercise.

Possible involvement of multidrug-resistance-associated protein (MRP) gene expression in spontaneous drug resistance to vincristine, etoposide and adriamycin in human glioma cells.

The multidrug-resistance phenotype in human tumors is partly associated with over-expression of the 170 kDa-P-glycoprotein encoded by the multidrug-resistance-1 (MDR1) gene. Another related, but non-P-glycoprotein, multidrug-resistance-associated protein (MRP) gene encodes a 190 kDa-membrane ATP-binding protein. Glioblastoma multiforme is a highly malignant primary neoplasm of the central nervous system which is refractory to anti-cancer chemotherapy, but the mechanism underlying this drug resistance is unknown. Out of glioma cell lines, 2, namely IN500 and T98G, which had elevated MRP mRNA levels, showed the highest resistance to multiple anti-cancer agents such as etoposide, vincristine and adriamycin, and decreased intracellular accumulation of etoposide. In the remaining 5 cell lines, various degrees of sensitivity to adriamycin and etoposide appeared to correlate with their respective MRP mRNA levels. Our study proposes that MRP may be involved in spontaneous multidrug resistance in human gliomas.