Search

Detection of resistance to drugs for Mycobacterium tuberculosis takes about two months from the sample collection using culture-based methods. To test a rapid method for detection of resistance for five antituberculosis drugs using DNA microarray and to examine its potential for clinical use, we employed a DNA microarray for detection of seven mutations genes related to resistance of five kinds of antituberculous drugs using Mycobacterium tuberculosis DNA isolated from sputum. The results of microarray analysis were compared with the results of a standard culture method of Lowenstein-jensen drug sensitivity testing system. DNA microarray analysis showed a high sensitivity (>90%) for all five drugs. Specificity of rifampicin and ethambutol were nearly 90%, however specificity of isoniazid (60%) and kanamycin (67%) were not enough. The amount of Mycobacterium tuberculosis DNA required for microarray analysis corresponded to at least 1–9 Acid-Fast Bacilli per 10 fields by carbolfuchsin staining. DNA microarray analysis appears to be useful for estimation of drug resistances, nevertheless its limitations. To minimize misunderstanding, it is necessary to confirm the number of bacilli in the sputum, and culture method is needed for comparison when use the PCR-based array system.

Antipituitary antibody (APA) has been reported to be detected in patients with autoimmune thyroid disease. Type 2 iodothyronine deiodinase (D2) is expressed in both pituitary gland and thyroid gland. We studied the association of APA and D2 peptide antibody in patients with autoimmune thyroid disease. Rat pituitary gland homogenate and D2 peptide were used as antigens in the present study. APA and D2 peptide antibodies were measured by enzyme-linked immunosorbent assay (ELISA) in sera obtained from 42 patients with Hashimoto's disease, 26 patients with Graves' disease and 70 healthy control subjects. Moreover, D2 activity precipitation assay was performed in some patients with Hashimoto's disease. APA and D2 peptide antibody were elevated in patients with Hashimoto's disease and patients with Graves' disease, compared with control subjects. APA was positive in 32.4% (22/68), D2 peptide antibody was positive in 26.5% (18/68) of patients with autoimmune thyroid disease. APA was positive in 31.0% (13/42) of patients with Hashimoto's disease and 34.6% (9/26) of patients with Graves' disease. D2 peptide antibody was positive in 26.2% (11/42) of patients with Hashimoto's disease and 26.9% (7/26) of patients with Graves' disease. D2 peptide antibody was correlated with APA in patients with autoimmune thyroid disease. Moreover, precipitation of D2 activity was increased in some patients with Hashimoto's disease including a patient who also had idiopathic diabetes insipidus, and was correlated with D2 peptide antibody. These results suggest that D2 antibody may be associated with APA in patients with autoimmune thyroid disease.

Our previous reports showed detection of antipituitary antibodies (APA) by immunoblot analysis and enzyme linked immunosorbent assay (ELISA) by using rat pituitary tissue as antigen in patients with certain endocrine disorders. In the present report, we evaluated APA by using our immunoblot and ELISA technique in 76 patients with various pituitary disorders. The prevalence of a 22 kDa band of APA detected by immunoblot was found to be significantly higher (P

We report a 61-year old male patient with panhypopituitarism complicated with asymptomatic primary biliary cirrhosis (PBC). T1-weighted magnetic resonance imaging demonstrated high intensity of the anterior pituitary gland. There was no mass lesion or enlargement of the pituitary gland or the stalk. Immunoblot analysis of the patient's sera with rat pituitary antigens revealed a band with a molecular size of 22 kD. Anti-M2 mitochondrial antibody has been consistently positive for five years. Liver biopsy revealed portal hepatitis with periportal infiltration of the inflammatory cells. This is the first case report of autoimmune hypophysitis complicated with asymptomatic PBC.

The volatile components from the thalloid liverwort, Marchantia paleacea subsp. diptera were investigated by HS-SPME-GC-MS analysis. The monocyclic monoterpene aldehyde, perillaldehyde was identified for the first time as the major component and its content was about 50% of the volatiles, along with β-pinene, limonene, β-caryophyllene, α-selinene and β-selinene as minor volatiles. Using MD (Multi-dimensional) GC-MS analysis equipped with a chiral column as the second column, the chirality was determined of both perillaldehyde and limonene, which was considered as the precursor of perillaldehyde,. Both compounds were ( S)-(-)-enantiomers (over 99.0 %) and ( R)-enantiomers (less than 0.5 %). This is the first report of the existence of perillaldehyde in liverworts.

OBJECTIVE To evaluate the clinical significance of the presence of anti-pituitary antibodies (APAs) in patients with NIDDM or IDDM and to examine the relationship of APAs to GAD antibodies (GADAs). RESEARCH DESIGN AND METHODS Serum samples were obtained from patients with NIDDM and IDDM. APAs, determined by Western blot analysis, and GADAs, determined by radioimmunoassay, were detected in the patients' sera and control sera. Urinary levels of C-peptide (U-CPR) were measured. RESULTS The prevalence of APAs was significantly higher in patients with NIDDM (24.2%) or IDDM (56.8%) than in healthy control subjects (6%). In patients with NIDDM, the levels of U-CPR were significantly lower, and the prevalence of insulin deficiency was higher in APA+ patients than in APA− patients. CONCLUSIONS This is the first study to demonstrate that the prevalence of APAs is increased in patients with NIDDM and IDDM. The presence of APAs may be related to reduced secretion of insulin in NIDDM patients.

The antituberculous drug isoniazid (INH) is acetylated by N-acetyltransferase 2 (NAT2), and the frequency of INH-induced hepatotoxicity is determined by the NAT2 genotype. NAT2 genotyping is not done routinely in hospital laboratories because of its difficulty. Use of microarrays for research is becoming common and its expectations of clinical application are increasing. In this study, we attempted to develop an easier method of NAT2 genotyping for clinical use. We devised a novel oligonucleotide-based DNA microarray for NAT2 genotyping using a recently developed technique for attaching oligonucleotide probes to poly carbodiimide-coated glass slides, which achieves a stronger hybridization signal and better specificity than the more widely used aminosilane-coated slides. To assess the validity of this microarray, four clones with NAT2 mutations and DNA from 42 tuberculosis patients were investigated by the microarray method and by restriction fragment length polymorphism analysis. The results of genotyping by these two methods were in agreement. Analysis of the relationship between the NAT2 phenotype determined by the DNA microarray and the risk of INH-induced hepatotoxicity revealed that slow acetylators had a significantly higher risk. These findings suggest that our microarray may be clinically useful for predicting drug reactions to INH.

The aim of the present study was to investigate the prevalence of antipituitary antibodies (APA) in patients with autoimmune thyroid disease as determined by Western blot analysis and enzyme-linked immunosorbent assay (ELISA). Results by Western blot analysis showed positivity for APA in serum of 22.4% of patients with Graves' disease (n=143, p

The clinical significance of anti-pituitary antibodies (APA) was examined by enzyme-linked immunosorbent assay (ELISA) in individuals with non-insulin dependent diabetes mellitus (NIDDM). Serum samples were obtained from 150 NIDDM patients and 45 normal subjects. Urinary C-peptide (U- CPR) was also measured for the NIDDM patients. APA-positive serum was incubated with porcine pancreas, liver, kidney, or spleen powder and analyzed by immunoblot. The prevalence of APA was found to be significantly (P

The identification of pituitary antigens recognized by human antipituitary antibodies (APAs) is important in evaluating the pathophysiology of multiendocrine disorders linked to autoimmune factors. However, there is no convenient method for the quantitative analysis of circulating APAs. This study reports the development of an enzyme-linked immunosorbent assay (ELISA) for the detection of APAs. APAs were measured by ELISA and confirmed by Western blot analysis in sera from patients with endocrine disorders. APAs were detected frequently in patients with autoimmune thyroiditis, insulin-dependent diabetes mellitus (IDDM), or pituitary dwarfism. Circulating APAs were detected in 18% of patients with autoimmune thyroiditis. Confirmation by Western blot revealed positivity for APAs in the serum of 36% of patients with Hashimoto disease and in 29% of patients with Graves disease. Notably, 39% of patients with IDDM were also positive for APAs by ELISA. The identification of APAs by ELISA may be useful in evaluating autoimmune mechanisms involved in patients with multiendocrine disorders.

Antipituitary antibodies have been reported to exist in sera of patients with autoimmune endocrine disorders. In order to investigate the pituitary antigens recognized by antipituitary antibodies, we studied the autoantigens in rat pituitary membrane and cytosolic fractions recognized by human antipituitary antibodies and anti-human pituitary hormone antibodies. Sera from 6 patients which showed positive antipituitary antibodies by immunofluorescence methods were studied. Each serum identified some proteins with 14.5, 22, 47, 49, 65, 84 and 97.5 kDa. Anti-GH antibodies and anti-PRL antibodies identified a positive band with 22 kDa, suggesting that anti-GH and anti-PRL antibodies may be present in patients' sera. These results indicate that Western blot analysis of rat pituitary antigens recognized by human antipituitary antibodies is a useful method to elucidate the pathophysiology of autoimmune endocrine disorders.

Thin films of amorphous Si3N4 were prepared by the rf-sputtering method, and the effects of titanium and chlorine additives on its crystallization were examined. When Ti-doped amorphous Si3N4 was heated, TiN precipitated at >1100°C; the TiN precipitates promoted the conversion of amorphous Si3N4 to β-Si3N4. Chlorine led to preferential conversion of amorphous Si3N4 to α-Si3N4.

The clinical figures and therapeutic treatment of a young male AIDS patient complicated with severe interstitial pneumonitis are described.The diagnosis was based on clinical symptoms and laboratory tests, including a lowered OKT4/T8 ratio (0.07) and the Western-Blot test. Antibodies to the virus reverse transcriptase (p61p24) had declined strongly, indicating the onset of AIDS. Necessary care taken for preventing the hospital employees from the transmission of the virus infections in the workplace are also described.A transmissible strain of human immunodeficiency virus (HIV) was isolated from the peripheral blood, and was identified by serological tests and electron microscopy.

The prevalence of persons antibody-positive for adult T cell leukemia virus-associated antigen (ATLA) or human immunodeficiency virus (HIV) was determined in 1, 815 blood donors and 3, 511 patients (total 5, 326) between April 1986 to February 1988.Among 1, 815 blood donors (healthy persons), 5 (0.28%) were confirmed to be sero-positive for ATLA by indirect immunofluorescence and Western-blot tests. None were positive for HIV.Among 2, 848 serum specimens of patients analyzed in the immunological laboratory of Gunma University Hospital, 26 (0.91%) were antibody-positive for ATLA and 13 (0.46%) for HIV. Twelve of the 13 anti-HIV antibody-positive patients had hemophilia, the other positive patient was a young male AIDS patient (non-Japanese) who had severe interstitial pneumonia.Among 663 patients who visited a venereal disease clinic, 2 (0.30%) were found to be positive for ATLA, but none were positive for HIV.