FUT2 non-secretor status is associated with Type 1 diabetes susceptibility in Japanese children.

AIM: To examine the contribution of the FUT2 gene and ABO blood type to the development of Type 1 diabetes in Japanese children. METHODS: We analysed FUT2 variants and ABO genotypes in a total of 531 Japanese children diagnosed with Type 1 diabetes and 448 control subjects. The possible association of FUT2 variants and ABO genotypes with the onset of Type 1 diabetes was statistically examined. RESULTS: The se2 genotype (c.385A>T) of the FUT2 gene was found to confer susceptibility to Type 1A diabetes in a recessive effects model [odds ratio for se2/se2, 1.68 (95% CI 1.20-2.35); corrected P value = 0.0075]. CONCLUSIONS: The FUT2 gene contributed to the development of Type 1 diabetes in the present cohort of Japanese children.

Variants associated with autoimmune Type 1 diabetes in Japanese children: implications for age-specific effects of cis-regulatory haplotypes at 17q12-q21.

AIMS: The aim of this study was to clarify the significance of previously reported susceptibility variants in the development of autoimmune Type 1 diabetes in non-white children. Tested variants included rs2290400, which has been linked to Type 1 diabetes only in one study on white people. Haplotypes at 17q12-q21 encompassing rs2290400 are known to determine the susceptibility of early-onset asthma by affecting the expression of flanking genes. METHODS: We genotyped 63 variants in 428 Japanese people with childhood-onset autoimmune Type 1 diabetes and 457 individuals without diabetes. Possible association between variants and age at diabetes onset was examined using age-specific quantitative trait locus analysis and ordered-subset regression analysis. RESULTS: Ten variants, including rs2290400 in GSDMB, were more frequent among the people with Type 1 diabetes than those without diabetes. Of these, rs689 in INS and rs231775 in CTLA4 yielded particularly high odds ratios of 5.58 (corrected P value 0.001; 95% CI 2.15-14.47) and 1.64 (corrected P value 5.3 × 10-5 ; 95% CI 1.34-2.01), respectively. Age-specific effects on diabetes susceptibility were suggested for rs2290400; heterozygosity of the risk alleles was associated with relatively early onset of diabetes, and the allele was linked to the phenotype exclusively in the subgroup of age at onset ≤ 5.0 years. CONCLUSIONS: The results indicate that rs2290400 in GSDMB and polymorphisms in INS and CTLA4 are associated with the risk of Type 1 diabetes in Japanese children. Importantly, cis-regulatory haplotypes at 17q12-q21 encompassing rs2290400 probably determine the risk of autoimmune Type 1 diabetes predominantly in early childhood.

Del(18)(q12.2q21.1) syndrome: a case report and clinical review of the literature.

The terminal deletion of the long arm of chromosome 18 is relatively common among cytogenetic abnormalities, which occur incidentally in approximately 1 in 40,000 live births. Proximal interstitial deletions of the long arm of chromosome 18 are less frequent. The critical region on chromosome 18 of this syndrome is del(18)(q12.2q21.1) and has recently been recognized as a new clinical entity. We describe a 8-year-old boy with developmental delay, obesity, and epilepsy, with characteristic facial anomalies in whom G-banding chromosome analysis revealed a unique karyotype of 46, XY, del(18)(q12.2q21.1). The patient was diagnosed with interstitial deletion chromosome 18q-syndrome. He received weight control therapy from a medical dietitian. For his epilepsy, he was administered oral carbamazepine at 4 mg/kg/day. At age six, he entered a special needs elementary school. After entering school, he often showed hyperkinesis, and was diagnosed with attention deficit hyperactivity disorder with mild mental retardation. Because patients with only del(18)(q12.2q21.1) have no serious associated malformations, physicians should be aware that even adult patients may have 18q-syndrome. Therefore, if epilepsy occurs in patients with minor facial anomalies, psychomotor retardation, obesity, and the possibility of 18q-syndrome with del(18)(q12.2q21.1) should be kept in mind, and chromosome testing should be performed.

Phenotypic features, androgen receptor binding, and mutational analysis in 278 clinical cases reported as androgen insensitivity syndrome.

Androgen insensitivity syndrome (AIS) is the most common single entity that results in male under-masculinization, but large cohort studies of AIS have rarely been performed. Over the last decade, nationwide cooperation between pediatric endocrinologists in the United Kingdom has allowed the creation of a database of cases of intersex and ambiguous genitalia where detailed clinical information on every notified case has been collected via a questionnaire. Among the 816 entries recorded by January 1999, there were 105 clinically diagnosed cases of complete AIS (CAIS) and 173 cases of partial AIS (PAIS). A masculinization score was devised by scoring the external phenotype, and a score of 12 represented normal masculinization. Androgen receptor (AR) binding was determined by studying binding capacity (Bmax) and receptor affinity (K(d)), and cases were classified as either zero, abnormal, or normal binding. Mutation screening of all eight exons of the AR gene was performed by single-strand conformational polymorphism analysis, followed by direct DNA sequencing. All cases of PAIS presented within the first month of birth. The median age at presentation of children with CAIS was 1 yr (P10,P90: 0.1,10.4). The testes were palpable in the labioscrotal folds or the inguinal region in 77% and 41% of cases of CAIS and PAIS, respectively. There was marked overlap between the masculinization score of those children with PAIS reared as girls [2.5(P10,P90:1, 6)] and those reared as boys [3(P10,P90:2, 7.5)]. Gonadectomy was performed prepubertally in 66% and postpubertally in 29% of the cases of CAIS. The median age of the latter group was older at 14 yr (P10,P90:0.1,18). No cases of malignancy or carcinoma in situ were reported in the 121 cases of AIS where histology results were available. Biochemical endocrine investigations were reported to have been performed in a greater number of cases of PAIS than CAIS (98% vs. 48%). AR binding was abnormal in 44 of 51 (86%) and 40 of 113 (35%) cases of CAIS and PAIS, respectively. Zero binding was encountered in 29 of 43 (67%) and 1 of 55 (2%) cases of CAIS and PAIS, respectively. Mutational analysis of the AR gene, performed in 102 index cases was positive in 57 of 69 (83%) cases of CAIS and 12 of 43 (28%) cases of PAIS. In 24 of these cases, the mutation identified was novel. The mutations in PAIS cases were all missense, whereas in CAIS the mutations were more diverse. AR binding was only normal in 3 of 69 mutation-positive cases. In the PAIS group, mutation-positive cases had a significantly higher Kd and Bmax compared to the mutation negative cases. The clinical diagnosis of AIS can be confirmed in a significant number of cases by a combination of androgen-binding studies and mutational analysis. There is some correlation between the phenotypic features and the abnormalities discovered on mutational analysis of the AR gene, but there is a need to improve this further by developing optimal bioassays of AR function. The phenotypic heterogeneity among clinically diagnosed cases of AIS emphasizes the need for appropriate comprehensive evaluation of male under-masculinization.

Localization of glutathione and induction of glutathione synthesis-related proteins in mouse brain by low doses of gamma-rays.

First, we determined the cerebral localization of reduced glutathione (GSH) in normal mice by means of autoradiography using 99mTc-meso-hexamethyl propylene oxime. A highly specific localization of GSH in the cerebellum and hippocampus was observed. Secondly, we measured the elevation of GSH level in the brain after low-dose gamma-irradiation. The cerebral GSH levels increased soon after irradiation with 50 cGy of gamma-rays, reaching a maximum at 3 h post-treatment, then remaining significantly higher than that of the non-irradiated control until 12 h and returning to the control level by 24 h. Thirdly, we examined the induction of the activities and the mRNAs of proteins involved in the synthesis and regeneration of GSH in the brain of mice subjected to low-dose gamma-ray irradiation. The level of mRNA for gamma-glutamylcysteine synthetase was significantly increased at 0.5 h, and remained high until 2 h post-irradiation (50 cGy). The level was transiently lowered to the non-irradiated control level at 3 h and slightly increased again after 6 h post-irradiation. gamma-Glutamylcysteine synthetase activity was significantly increased 3 h after irradiation, and remained high up to 24 h post-irradiation. As for glutathione reductase, the mRNA level was increased at 0.5 h, and peaked strongly at 2 h, while the enzyme activity was significantly increased at 6 h after irradiation, and continued to increase up to 24 h. The level of mRNA for thioredoxin, which contributes to GSH biosynthesis by supplying cysteine to the de novo pathway, peaked between 0.5 h and 2 h post-irradiation, and rapidly declined thereafter. The content of thioredoxin showed a transient decrease immediately after irradiation, but was then remarkably elevated, reaching a maximum at 3 h, and thereafter declining sharply. These results indicate that the increase in endogenous GSH in mouse brain soon after low-dose gamma-ray irradiation is a consequence of the induction of GSH synthesis-related proteins and occurs via both the de novo synthesis and the regeneration pathways.

The effect of NaF on the bacterial production of polysaccharide and subsequent adsorption on hydroxyapatite.

The affinity of some bacterial polysaccharides for hydroxyapatite was investigated. Water insoluble polysaccharide production by bacteria was inhibited by the presence of NaF in the growth medium, while the water soluble fraction was unaffected. The adsorption of the bacterial polysaccharide on hydroxyapatite decreased with decreasing levels of the water insoluble fraction. It is suggested that NaF can inhibit bacterial attachment and caries by interfering with bacterial attachment through reducing the production of water insoluble polysaccharide.

Determination of estrogen 3-sulfates in biological fluids of mammary tumor-bearing rats by radioimmunoassay.

Determination of estradiol 3-sulfate (E2 3-S) and estriol 3-sulfate (E3 3-S) in plasma, urine or tumor tissues of mammary tumor-bearing rats were performed using the superior radioimmunoassay (RIA) system. The plasma level of E2 3-S after tumorigenesis was found to be about one-third of the normal level. As to E3 3-S, the levels in urine were significantly high both before and after tumorigenesis. Before that, the average level was about 1.5 times, and after that, about 2.5 times as high as the normal level. In tumor tissues, an extremely high level of E3 3-S (399.6 +/- 113.9 pg/g tissue) was determined.

Specific imaging of hormone-dependent mammary carcinoma in nude mice with [131I]-anti-estriol 3-sulfate antibody.

We tried to put the estrogen metabolite to use in tumor imaging. The antibody against estriol 3-sulfate (E3 3-S), which was one of the major metabolites of estrogen in hormone-dependent mammary carcinoma, was prepared and the tissue distribution and imaging of human breast carcinoma with anti-E3 3-S antibody (Ab) were studied in nude mice. In hormone-dependent breast carcinoma, MCF-7,-bearing nude mice, [125I] anti-E3 3-S Ab localized in tumor with the percentage injected dose/g of 9.29 +/- 3.01 (mean +/- SD). This value was significantly high compared with that in hormone-independent breast carcinoma, MDA-MB-231,-bearing nude mice. At 72 h after the administration of [125I]anti-E3 3-S Ab to MCF-7 bearing mice, tumor/blood, tumor/liver and tumor/muscle ratios were 0.49, 5.02 and 6.83, respectively. These ratios were supposed to be enough for imaging. In radioimmunoscintigraphy, a MCF-7 tumor was clearly visualized at 120 or 168 h post-injection of [131I]anti-E3 3-S Ab.

Comparative study of intact A7 MoAb and F(ab')2 fragments for radioimmunoimaging of human colon cancer in nude mice.

Differences of pharmacokinetics and tumor imaging ability between intact monoclonal antibody A7 (A7 MoAb) and F(ab')2 fragments were studied in human colon cancer (LS-174T)-bearing nude mouse. First of all, we examined the yield and the immunoreactivity of F(ab')2 fragments after treatment with ficin as a function of time. The yield of F(ab')2 fragments reached about 50% after ficin treatment for 8 h, and the F(ab')2 retained about 80% of the immunoreactivity of the corresponding MoAb. Longer digestion with ficin produced smaller fragments (less than 92 kDa) with a lower yield and most of the immunoreactivity was lost. In pharmacokinetics studies, the F(ab')2 was preferentially taken up by the tumor, was cleared more rapidly from the blood circulation and seemed to have less non-specific tissue binding than intact A7 MoAb. In addition, the tumor image obtained at an early time using 131I-F(ab')2 was much superior in quality to that with intact 131I-A7 MoAb. The use of F(ab')2 fragments may be effective for tumor diagnosis and therapy.

Radioiodination of glycoprotein-conjugated liposomes by using the Bolton-Hunter reagent and biodistribution in tumor-bearing mice.

We have developed a suitable radiolabeling method for our new type of glycoprotein-liposome conjugate (GCL), in order to investigate its potential utility as a drug carrier that can target the cellular functions of carbohydrate-binding proteins. In order to obtain radiolabeled GCL with high labeling efficiency, we introduced p-hydroxyphenylpropyl groups into the liposome membrane through the amine moiety of a constitutive phospholipid, dipalmitoylphosphatidylethanolamine (DPPE) by using Bolton-Hunter reagent (BHR). Radioiodination of the introduced tyrosyl groups was performed by the Chloramine-T method. The labeling efficiency of the BHR-treated liposome conjugate was high in comparison with that of the BHR-untreated liposome conjugate. An in vitro inhibition study showed that the binding affinity of 125I-labeled BHR-treated GCL (125I-F3S-BH) with lectin was twice as high as that of untreated conjugate (125I-F3S). The biodistribution of 125I-F3S-BH in mice was considerably different from that of 125I-F3S. 125I-F3S-BH was more rapidly taken up by the liver and was more rapidly excreted from the liver than 125I-F3S. Moreover, 125I-F3S-BH accumulated more rapidly into the kidneys, which resulted a lower radioactivity in the blood circulation at an earlier time point than in the case of 125I-F3S. The characteristics of tumor accumulation of 125I-F3S-BH and 125I-F3S were similar to those in blood. If F3S is to be employed as an in vivo targeting ligand in biodistribution studies, BHR would be a suitable tool for radiolabeling because it allows GCL to retain the biological activity and characteristics of the unmodified conjugate.

Differences between A7 antibody and anti-CEA antibody.

The specificity of A7 monoclonal antibody (A7 MoAb), raised against a human colon cancer, was investigated in detail and compared with that of anti-CEA MoAb. Firstly, the biodistributions of radiolabeled A7 MoAb and anti-CEA MoAb were examined in human colon cancer (LS-174T)-, glioblastoma- and lung cancer (Lu-65)-bearing nude mice. 125I-A7 MoAb was highly concentrated into LS-174T and also, to a lesser extent, in glioblastoma, whereas 125I-anti-CEA MoAb was significantly taken up only by LS-174T. Both of these antibodies were taken up at only very low concentrations into Lu-65. Secondly, in vitro binding studies of 125I-A7 MoAb and 125I-anti-CEA MoAb with LS-174T cells and glioblastoma cells revealed high binding activity of these MoAbs to LS-174T cells, though A7 MoAb had a much higher affinity than that of anti-CEA MoAb. Neither of them showed high affinity for glioblastoma cells. Thirdly, competitive binding assay using A7 MoAb and anti-CEA MoAb to LS-174T cells showed that the binding of each 125I-MoAb was not inhibited by the other MoAb. Finally, in radioimmuno-imaging studies of LS-174T- and glioblastoma-bearing mice with 131I-A7 MoAb and -anti-CEA MoAb, both tumors were clearly visualized with the former, while the latter visualized only LS-174T. The A7 MoAb is clearly different from anti-CEA MoAb and may be useful for the in vivo radioimmunodetection and treatment of colon cancer.

Usefulness of (99m)Tc-d,l-HMPAO for estimation of GSH content in tumor tissues.

To investigate whether [(99m)Tc]-hexamethyl propyleneamine oxime ([(99m)Tc]-HMPAO) is applicable for evaluating glutathione (GSH) localization in tumor, the difference of distribution between [(99m)Tc]-d,l- and meso-HMPAO was studied using a mouse tumor model. Biodistribution of [(99m)Tc]-d,l- or meso-HMPAO was studied in GSH-depleted and control Ehrlich tumor-bearing mice. GSH levels in tumors in GSH-depleted and control mice were measured in another set of mice. The uptake of [(99m)Tc]-d,l-HMPAO in tumor was significantly decreased by the diethyl maleate (DEM) treatment. On the other hand, the DEM treatment increased the accumulation of [(99m)Tc]-meso-HMPAO in tumor. Meanwhile, the content of GSH was lowest in tumor among the tissues tested and decreased in a manner similar to other tissues on preloading of DEM. [(99m)Tc]-d,l-HMPAO may be useful for estimating the GSH status in a certain tumor and thereby contribute to the diagnosis of anticancer therapy.

Phenotyping of lymphocyte subsets in the vascular and epithelial lesions of a cow with malignant catarrhal fever.

Surface marker analysis of lymphoid cells infiltrating the vascular and epithelial lesions of a cow with malignant catarrhal fever (MCF) was conducted by immunohistochemistry using ten monoclonal antibodies. The majority of lymphoid cells in these lesions had BoCD8, BoCD6 or BoCD2, but they rarely possessed N-cell (BoCD5+/BoCD4-/BoCD8-, non-T non-B) markers. Similar reactivity was seen in lymphoid cells of perivascular infiltrates in the liver, heart and brain, and in T-dependent areas of lymph nodes. These results suggest involvement of cytotoxic T-lymphocytes in the pathogenesis of MCF.

Inhibition of phosphatidylinositol kinase by toyocamycin.

We have screened toyocamycin as an inhibitor of phosphatidylinositol kinase. It inhibited the enzyme of A431 cell membrane with an IC50 of 3.3 micrograms/ml. Adenosine and formycin A also inhibited the enzyme, but other 6 related nucleosides did not. Although orobol and 2,3-dihydroxybenzaldehyde that inhibit phosphatidylinositol kinase inhibited in situ phosphatidylinositol turnover, toyocamycin did not.

Anxiety and dental caries.

It is accepted that emotional stress causes various kinds of degenerative physiological changes. We have assumed that it is important to study the relationship between anxiety and dental caries. Anxiety and personality of children were investigated and oral examinations and questionnaires about their life style were performed. There was statistically a significant relationship between dental caries and anxiety which may be overridden by keeping regular hours. To prevent dental caries, we conclude that more attention should be paid to psychosomatic responses which may cause dental caries.

Ovarian cysts in precocious puberty.

We describe a 7-year-old girl with precocious puberty in whom a single large cyst (5 cm) and several small cysts (8-10 mm) in the single remaining ovary were detected by the ultrasound examination. Endocrinological examinations confirmed the diagnosis of central precocious puberty. Pathologic findings after the removal of the cystic lesions revealed that the large cyst was derived from degenerated follicular cysts and the small cysts were identical to follicular cysts: all were considered to have been formed by gonadotropin stimulation. In general, surgical removal of an ovarian follicular cyst in central precocious puberty is inappropriate. However, in this unusual patient who had a degenerated large cyst, surgery seemed to be appropriate because of a previously removed teratoma in the contralateral ovary.

[New curriculum of behavioral dentistry].

Faculty of Dentistry, Tokyo Medical and Dental University first introduced New Curriculum of Behavioral Dentistry in Japan. This curriculum has been established in consideration of the current social environments and dental clinical practice in our country, in addition to reference to Curriculum Guidelines in the USA. When students' impressions and assessments on the courses were researched and analyzed, the majority were found to be satisfied with this curriculum although several problems to be solved were also presented at the same time, such as timing of derivering lectures.

[Medical interview with simulated patients at behavioral science in dentistry].

The purpose of this paper is to present an outline of student's practice of interviewing simulated patients at behavioral science in dentistry. This practice was initiated as part of a newly introduced behavioral science course at our school of dentistry, to enable students to acquire communication skills, comprehensive understanding, and a proper attitude vis-à-vis patients. Students as well as instructors involved in the practice evaluated it as highly relevant for clinical education. It is concluded that the development of such practices in dental education is a prerequisite for training students to dentists oriented toward patient-centered dental practice.