Influence of the polymorphism of the DUSP14 gene on the expression of immune-related genes and development of pulmonary tuberculosis.

Recently, a genome-wide screening identified a functional single-nucleotide polymorphism in dual-specificity phosphatase 14 gene (DUSP14), which was associated with pulmonary tuberculosis (TB) in a West African study. DUSP14 regulates T-cell proliferation and cytokine production in a negative way via dephosphorylation and inactivation of key signaling molecules. The aim of this study is to further explore the possible significance of the DUSP14 polymorphism. Total RNA was extracted from the whole blood of 109 healthcare workers (HCWs) in Vietnam and subjected to quantitative reverse-transcription PCR for DUSP14 and 20 immune-related genes. DUSP14 rs1051838 was genotyped in 502 new pulmonary TB patients and 506 healthy controls. Among disease-free individuals (HCWs), T-helper type-1 (Th1)-related genes, interferon-gamma receptor 2 (IFNGR2) and signal transducer and activator of transcription-1 (STAT1) mRNA levels significantly increased as the number of A alleles of rs1051838 increased, whereas the DUSP14 mRNA level tended to decrease. The AA genotype was associated with protection against active TB in younger patients (⩽45 years old, OR=0.63, 95% CI 0.44-0.90). Our results suggest that a low-expression genotype of DUSP14 accompanied by high transcript levels of Th1 immune-related genes may confer protection against early TB development.

Improvement of (R)-3-hydroxybutyric acid secretion during Halomonas sp. KM-1 cultivation with saccharified Japanese cedar by the addition of urea.

UNLABELLED: Japanese cedar (Cryptomeria japonica) is a major species in artificial Japanese forests. The Halomonas sp. KM-1 was recently isolated and found to grow effectively on saccharified Japanese cedar wood, resulting in the intracellular storage of poly-(R)-3-hydroxybutyric acid (PHB) under aerobic conditions. Under microaerobic conditions, the extracellular secretion of (R)-3-hydroxybutyric acid ((R)-3-HB) led to the degradation of intracellular PHB. In this study, the production of PHB and the secretion of (R)-3-HB using saccharified Japanese cedar were much improved in cultures that were grown in the presence of urea. The level of intracellular PHB production after 36 h under aerobic cultivation was 23·6 g l(-1) ; after shifting to microaerobic conditions for 24 h, the (R)-3-HB concentration in the medium reached 21·1 g l(-1) . Thus, KM-1 efficiently utilizes saccharified Japanese cedar to produce PHB and secretes (R)-3-HB, making it a practical candidate for use in the industrial production of (R)-3-HB. SIGNIFICANCE AND IMPACT OF THE STUDY: Japanese cedar is a major species grown in artificial Japanese forests, and its thinning is crucial for the health of artificial forests and the Japanese economy. Halomonas sp. KM-1 grew effectively on saccharified Japanese cedar wood, resulting in intracellular storage of poly-(R)-3-hydroxybutyric acid (PHB) under aerobic conditions. Under microaerobic conditions, extracellular secretion of (R)-3-hydroxybutyric acid ((R)-3-HB) caused intracellular PHB degradation. (R)-3-HB is a chiral compound that is useful in the chemical, health food and pharmaceutical industries. The production of PHB and secretion of (R)-3-HB using saccharified wood was dramatically improved, which may positively affect its future industrial production.

Association analysis of susceptibility candidate region on chromosome 5q31 for tuberculosis.

Chromosome 5q31 spans the T helper (Th) 2-related cytokine gene cluster, which is potentially important in Th1/Th2 immune responses. The chromosome 5q23.2-31.3 has been recently identified as a region with suggestive evidence of linkage to tuberculosis in the Asian population. With the aim of fine-mapping a putative tuberculosis susceptibility locus, we investigated a family-based association test between the dense single nucleotide polymorphism (SNP) markers within chromosome 5q31 and tuberculosis in 205 Thai trio families. Of these, 75 SNPs located within candidate genes covering SLC22A4, SLC22A5, IRF1, IL5, RAD50, IL13, IL4, KIF3A and SEPT8 were genotyped using the DigiTag2 assay. Association analysis revealed the most significant association with tuberculosis in haplotypes comprising SNPs rs274559, rs274554 and rs274553 of SLC22A5 gene (P(Global)=2.02 x 10(-6)), which remained significant after multiple testing correction. In addition, two haplotypes within the SLC22A4 and KIF3A region were associated with tuberculosis. Haplotypes of SLC22A5 were significantly associated with the expression levels of RAD50 and IL13. The results show that the variants carried by the haplotypes of SLC22A4, SLC22A5 and KIF3A region potentially contribute to tuberculosis susceptibility among the Thai population.

Genome-wide SNP-based linkage analysis of tuberculosis in Thais.

Tuberculosis, a potentially fatal infectious disease, affects millions of individuals annually worldwide. Human protective immunity that contains tuberculosis after infection has not been clearly defined. To gain insight into host genetic factors, nonparametric linkage analysis was performed using high-throughput microarray-based single nucleotide polymorphism (SNP) genotyping platform, a GeneChip array comprised 59 860 bi-allelic markers, in 93 Thai families with multiple siblings, 195 individuals affected with tuberculosis. Genotyping revealed a region on chromosome 5q showing suggestive evidence of linkage with tuberculosis (Z(lr) statistics=3.01, logarithm of odds (LOD) score=2.29, empirical P-value=0.0005), and two candidate regions on chromosomes 17p and 20p by an ordered subset analysis using minimum age at onset of tuberculosis as the covariate (maximum LOD score=2.57 and 3.33, permutation P-value=0.0187 and 0.0183, respectively). These results imply a new evidence of genetic risk factors for tuberculosis in the Asian population. The significance of these ordered subset results supports a clinicopathological concept that immunological impairment in the disease differs between young and old tuberculosis patients. The linkage information from a specific ethnicity may provide unique candidate regions for the identification of the susceptibility genes and further help elucidate the immunopathogenesis of tuberculosis.

Anti-arthritic effects of combined treatment with histone deacetylase inhibitor and low-intensity ultrasound in the presence of microbubbles in human rheumatoid synovial cells.

OBJECTIVE: The therapeutic effects of histone deacetylase (HDAC) inhibitor combined with ultrasound (US) (1 MHz, 10% duty factor, 0.1 or 0.2 W/cm(2)) in RA synovial fibroblasts (RASFs) were examined. METHODS: RASFs were isolated from rheumatoid synovial tissues obtained from patients with RA during total knee arthroplasty. RASFs were treated with an HDAC inhibitor, trichostatin A (TSA), with or without US. Cell viability was estimated using the Trypan blue dye exclusion test and cell cycle was examined by flow cytometry using propidium iodide (PI) staining. Gene expression of cell cycle-related genes cyclin D, cyclin A, cyclin B and p21(WAF1/Cip1) was analysed by semi-quantitative RT-PCR. Detection of apoptosis was examined by flow cytometry using annexin V-FITC and PI staining. Microarray analysis was carried out to profile gene expression of inflammation-related genes. RESULTS: Dose-dependent decreases in cell viability, cell cycle arrest and apoptosis in RASFs due to TSA were observed. US treatment in the presence of microbubbles increased cellular uptake, but did not induce cell cycle arrest or apoptosis. The combination of TSA and US modulated cell cycle-related gene expression and significantly decreased S phase cells and increased G(2)-M phase cells. US also further enhanced TSA-induced RASF apoptosis and regulated expression of inflammation-related genes. CONCLUSIONS: HDAC inhibitor in combination with US effectively reduces cell viability and induces apoptosis in RASFs. The combination therapy could be useful to control synovial proliferation and inflammation, since US can be easily applied to targeted joints as local physiotherapy.

Enhancement of coxsackievirus B3 myocarditis in mice by lobenzarit disodium through inhibition of splenic pan T cells.

OBJECTIVE: The aim was to test the efficacy of the immune system modulator lobenzarit disodium in the treatment of coxsackievirus B3 myocarditis. METHODS: Two week old C3H/He mice were inoculated with 10(3) plaque forming units of coxsackievirus B3. Lobenzarit disodium, 25 mg.kg-1.d-1, was given subcutaneously daily on days 0-14 (experiment I; group 2) and days 14-28 (experiment II; group 4). Both treated groups were compared to infected controls for each experiment (groups 1 and 3). For the analysis of splenic lymphocyte subsets, additional mice in untreated and treated groups were killed on d 7, and the percentages of Thy 1.2 (CD3), L3T4 (CD4), Ly 2 (CD8) subsets were analysed by laser flow cytometry (experiment III). RESULTS: In experiment I, the survival rate in the lobenzarit treated group was significantly lower than in the controls (2/11 v 8/11). Cellular infiltration and myocardial necrosis in the lobenzarit group were more severe. Myocardial virus titres and serum neutralising antibody titres did not differ significantly between the two groups. In experiment II, the survival rate (7/9 v 13/13) and cardiac pathology between the two groups did not differ significantly. In experiment III, the percentage of the Thy 1.2 subset (CD3) in the treated group was significantly lower (p < 0.05) than in the control group, at 36.0(SD 2.9)% v 42.8(5.8)%. CONCLUSIONS: Lobenzarit disodium decreased splenic pan T cells and aggravated both clinical course and cardiac pathology in acute murine coxsackievirus B3 myocarditis.

Support for an association between HLA-DR1 and schizophrenia in the Japanese population.

An increase of HLA-DR1 has been observed in schizophrenia patients from the Japanese population. A decrease of DR4, which was reported in Caucasian patients, has also been found in some of the Japanese studies. This small study further investigated frequencies of HLA-DR1 and DR4 in unrelated Japanese patients with schizophrenia (n = 45) and healthy comparison subjects (n = 117). The number of patients possessing DR1 was higher (10 of 45, 22%) compared with the comparison group (11 of 117, 9.4%, P = 0.03). This may support the previous observation of an increased DR1 frequency in the Japanese patients. When the present data is combined with three previous studies, proportions of the Japanese subjects with DR1 were 98 of 588 schizophrenia patients (16.7%) vs. 93 of 942 comparison subjects (9.9%). However, no difference was observed in DR4 frequencies between the patients (51%) and comparison subjects (44%). Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:725-727, 2000.

Immunopharmacological evaluation of bucillamine in rheumatoid arthritis.

Seventy six patients with rheumatoid arthritis (RA) treated with bucillamine, a novel immunomodulating drug with no antiinflammatory effects, were evaluated in terms of the immunomodulating effects. Clinical symptoms and laboratory data, including lymphocyte subset changes, were followed for 12 or more months. Long-term administration of bucillamine markedly improved arthritis and suppressed the titre of rheumatoid arthritis haemagglutination. Flow cytometric analysis of peripheral blood T cell subsets showed that bucillamine induced a characteristic increase in the cell constituent ratio of CD4+CD45RA+ T cells. Decreases in CD4+CD45RO+ T cells, CD8+HLA-DR+ T cells and CD5+ B cells were considered characteristic features induced by bucillamine administration.

Mucoepidermoid carcinoma of the salivary gland in pleural fluid. A case report.

The cellular features of mucoepidermoid carcinoma of submandibular gland origin observed in pleural fluid are presented. The pleural fluid contained predominantly atypical spheroid cell clusters accompanied by numerous mesothelial cells. The cells had round nuclei with conspicuous nucleoli, coarsely granular chromatin and abundant cytoplasm with vacuoles. The cellular features of the malignant cells in the pleural fluid were correlated with the histology of the parent lesion.

Cytologic demonstration of Entamoeba histolytica using immunoperoxidase techniques. Report of two cases.

An indirect immunoperoxidase technique to detect Entamoeba histolytica in cell samples from patients suspected to have amebiasis is described. Using a rat antiserum specific for E. histolytica, the organism was clearly identified both in smears and in cell blocks. This immunoperoxidase technique seems to offer great possibilities for a specific, accurate and rapid identification of amebic infestation in diagnostic cytology.

[Antibiotic concentration during extracorporeal circulation (author's transl)].

In open surgery, ampicillin (Solcillin) or cephacetrile (Celtol), 2 g for patients weighing 20 kg or more, and 1 g for those below 20 kg, was administered, and the antibiotic concentrations in blood and urine were estimated during extracorporeal circulation, from the time of operation to the admission in an intensive care unit (Group A). In other group, a total circulation volume of 0.3 mg/ml of the antibiotic was administered, based on body weight of patients and priming volumes (Group B). In Group A, blood concentration of antibiotics was so variable that it was difficult to decide additional dosage. In Group B, comparatively definite concentrations were estimated in each case. In extracorporeal circulation for a long time, it is preferable to maintain the blood concentration of antibiotic at 50 micrograms/ml, by additional antibiotic administration 90 approximately 120 minutes after the beginning of the extracorporeal circulation. In patients with preoperative subacute bacterial endocarditis, the blood concentration should be kept over 100 micrograms/ml during extracorporeal circulation.

[Two cases of urachal carcinoma].

Two cases urachal carcinoma are presented. Both of them had complaints of macrohematuria. One patient received partial cystectomy, and the other patient had radical cystectomy, peritoneo-umbiliectomy, and ureterosigmoidstomy after CDDP and ADM intraarterial injection. The former patient was found to have distant metastasis 7 months after operation. The latter patient has been disease-free for 4 months. The most important point to cure urachal carcinoma is accurate staging and sufficient first treatment, especially pre-operative intraarterial chemotherapy.

Renal hemangioma.

Four cases of renal hemangioma are presented. Renal hemangioma is difficult to detect because this benign vascular tumor never demonstrates any abnormalities on renal arteriography. Computed tomography in two resected cases of cavernous hemangioma revealed a low-density mass without any enhance effect, while the others diagnosed by the selective renal arteriography demonstrated no abnormality on computed tomography. We postulate that both angiographic and computed tomographic appearances of the renal hemangioma could depend on its vascular components. Related reports are also reviewed.

[A case of soft renal calculi with xanthogranulomatous change].

A case of soft renal calculi with xanthogranulomatous change is reported. A 37-year-old female visited our hospital on February 4, 1992 complaining of frequency of urination and right lower abdominal pain. Under the clinical diagnosis of right renal calculi, extracorporeal shock wave lithotripsy was attempted, but no sign of destruction was observed. Right pyelolithotomy was performed on June 8, 1992. Several soft calculi were removed from the right renal pelvis. Microscopic examination showed that the calculi were surrounded by a cell layer including foam cells and giant cells. In addition, the calculi revealed positive reaction by several stainings (alcian-blue, PAS and muticarmin), which showed that the calculi were mainly composed of mucopolysaccharides. We discussed this disease in terms of symptoms, diagnosis, treatment and mechanism in comparison with previous reports.

[A case of renal lymphoma].

A 74-year-old man was admitted to our hospital, complaining of gross hematuria and complete urinary retention. Right nephrectomy and suprapubic prostatectomy were performed under general anesthesia. On the second postoperative day, acute renal failure developed and peritoneal dialysis was begun. But, unfortunately he died the next day. Pathological diagnosis was malignant lymphoma infiltrating to the perirenal and periadrenal adipose tissue.

Pulmonary Mycobacterium avium complex infection: association with NRAMP1 polymorphisms.

The present study aimed to elucidate risk factors for nonimmunocompromised pulmonary Mycobacterium avium complex (MAC) infection. Epidemiological data and variations of candidate genes for mycobacterial diseases were analysed in 111 patients with pulmonary MAC infection. Four polymorphisms of the human natural resistance-associated macrophage protein (NRAMP)1 gene, the 5'(GT)n, 469+14 G/C, D543N and the 3'untranslated region (3'TGTG) insertion/deletion, were genotyped using PCR-based methods. Fok I and Taq I polymorphisms of the vitamin D receptor gene and -221 X/Y and codon 54 A/B polymorphisms of the mannose binding lectin gene were also evaluated. Females were more susceptible to MAC infection mainly affecting the right middle lobe or lingular segment of the lung. Patients' residence at the onset of the disease was distributed evenly irrespective of a waterfront or city water supply system. As compared with homozygotes for major alleles of the D543N and TGTG insertion/deletion polymorphism of the NRAMP1 gene, heterozygotes containing minor alleles were less often observed in MAC cases than in controls. This genetic effect was more significant in patients without comorbidity but not in patients with comorbidity. Other polymorphisms did not show any association with the MAC infection. The human natural resistance-associated macrophage protein 1 gene might be involved in susceptibility to pulmonary Mycobacterium avium complex infection.