A novel HLA-B*35 allele, B*35:188, identified by sequence-based typing.

The novel allele B*35:188 allele showed a single nucleotide difference with B*35:96 at nt 347 T>C in exon 3.

Induction of MMP-13 expression by soluble human glucocorticoid-induced tumor necrosis factor receptor in fibroblast-like synovial cells.

OBJECTIVE: We tested the hypothesis that human glucocorticoid-induced tumor necrosis factor receptor (hGITR/TR11) expressed on the surface of activated CD4(+) T cells is responsible for up-regulating the production of matrix metalloproteinase (MMP)-13 by fibroblast-like synoviocytes (FLSs). METHODS: The level of MMP-13 was measured by Western blot and reverse transcriptase polymerase chain reaction (RT-PCR). Expressions of hGITR ligand (hGITRL) on the surface of FLSs and hGITR on the surface of human CD4(+) T cells were analyzed by flow cytometry and RT-PCR. Neutralizing antibodies (Abs) were used to block hGITRL and hGITR on the surface of FLSs and human CD4(+) T cells, respectively. Human CD4(+) T cells were cocultured with FLSs to facilitate interaction between hGITR on CD4(+) T cells and hGITRL on FLSs. RESULTS: Soluble hGITR (shGITR) stimulated FLSs to produce MMP-13, and blockade of hGITRL reduced this effect. Direct contact between activated CD4(+) T and FLSs also induced the production of MMP-13, and neutralization of hGITR on activated CD4(+) T cells during coculture decreased the amount of MMP-13 produced by FLSs. CONCLUSION: shGITR stimulated FLSs to produce MMP-13 via a signal through hGITRL. Direct contact between activated CD4(+) T cells and FLSs facilitated hGITR-hGITRL interaction, and resulted in inducing MMP-13. This effect may increase tissue destruction in chronic inflammation such as rheumatoid arthritis (RA).

Dyspnea and palpitation during pregnancy.

OBJECTIVES: Dyspnea and palpitation are common features of pregnancy. While several theories have been put forward to explain the etiology of gestational dyspnea and palpitation, there have been few systemic studies of its incidence, severity and time-course in a group of normal women. METHODS: We interviewed postpartum women, within 3 days after delivery, about dyspnea and palpitation. Separately from this interview, we performed 24-hour ECG monitoring for obstetric patients with palpitation before delivery. RESULTS: The subjects interviewed were 261 women, of whom 37.5 percent and 11.5 percent experienced dyspnea and palpitation, respectively. These symptoms had tendency to increase to term. The presence of arrhythmias could be documented in only 22% of patients having 24-hour Holter monitoring. CONCLUSION: Dyspnea and palpitation were common among normal pregnant women and had a tendency to increase to term.

Effects of oxygen on invertase expression in continuous culture of recombinant Saccharomyces cerevisiae containing the SUC2 gene.

The yeast SUC2 gene, cloned on a multicopy plasmid pRB58, was used to study the effect of oxygen on the invertase expression of the recombinant Saccharomyces cerevisiae. Glucose repression was not the only factor affecting the invertase expression. The results obtained from the single-stage continuous cultures under microaerobic conditions showed that invertase expression was also strongly dependent on oxygen availability, and moving from anaerobic to aerobic conditions led to a five-fold increase in specific invertase activity. However, the cell yields under anaerobic conditions were quite low compared to those under aerobic conditions. These opposite effects of oxygen on cell growth and gene expression offer a strategy for maximizing invertase productivity by a two-stage continuous culture. The first stage was operated at a low level of glucose, around 100 mg/l, under aerobic conditions in order to obtain a high yield of yeast biomass, and the second stage maintained anaerobic conditions with residual glucose levels of 50 mg/l to derepress and fully induce invertase expression. The two-stage continuous culture resulted in a 2.5-fold increase in invertase productivity over that of a single-stage continuous culture.

Requirement of MEF2D in the induced differentiation of HL60 promyeloid cells.

The regulatory role of MEF2 (myocyte enhancer binding factor 2) proteins in nonmuscle tissues has not been well characterized. We examined the expression of MEF2 family members, namely, MEF2A, -B, -C, and -D, in the differentiation of HL60 promyeloid cells and observed the remarkable increase in the expressions of MEF2A and MEF2D proteins during the differentiation process into monocytes. To examine the role of MEF2, we expressed a dominant-negative form of MEF2D, without its transactivation domain, in HL60 cells. When the HL60 cell line expressing the mutant MEF2D was induced to differentiate by VitD(3) treatment, cell surface expression of CD14 and the ability to reduce NBT, which are important characteristics of differentiated monocytes, were significantly decreased compared with control HL60 cells. These results show that MEF2D is required in the differentiation process along the monocyte/macrophage lineage,

Purification and characterization of cysteine protease from Pleurotus ostreatus.

Cysteine protease activity in mycelial culture increased 7.7-fold after fruit body formation in Pleurotus ostreatus, using the Leu pNA (LPNA) cleavage assay. The enzyme was purified from fruit bodies and its M(r) was 97,000 by gel filtration and 48,500 by SDS-PAGE, indicating that it is a dimer. The enzyme was sensitive to iodoacetic acid, p-chloromercuribenzoate, N-ethylmaleimide, and HgCl2. The sequence of the first 9 N-terminal amino acids of cysteine protease was ASGLXXAIL.

Increased prevalence of abnormal signal-averaged electrocardiograms in older patients who have hypertension with low diastolic blood pressure.

Greater mortality in hypertensive patients with the lowest diastolic blood pressure (DBP) values has been reported and may be partially explained by more fatal arrhythmias. The records of 135 patients were reviewed to determine whether the prevalence of an abnormal signal-averaged electrocardiogram (SAECG+), a risk marker for ventricular arrhythmias, was greater in patients who have hypertension with DBP < 85 mm Hg. SAECG+ was present in 31 (39%) of 80 patients with hypertension and 13 (24%) of 55 subjects with normotension p < 0.05. Hypertensive patients were more likely than normotensive subjects to have left ventricular hypertrophy (LVH, p = 0.01) and LV dysfunction (p < 0.01). In multivariate analysis only age and systolic dysfunction emerged as significant predictors of SAECG+. Among 68 hypertensive patients and for whom recent DBP data were available, SAECG+ was present in 18 of 37 with DBP < 85 mm Hg (group 1), 1 of 12 with DBP 85 to 94 mm Hg (group 2, p < 0.05 vs group 1), and 4 of 19 with DBP > 95 mm Hg (group 3, p < 0.05 vs group 1, p = 0.08 vs group 2). There were no significant differences between the three groups of hypertensive patients for coronary artery disease, LVH, systolic dysfunction, or left-sided cardiac chamber enlargement. Multivariate analysis in hypertensive patients indicated that age and DBP were each independently predictive of SAECG+, whereas the indexes of heart disease were not. Within group 1, those with SAECG+ were significantly older (68 +/- 1 vs 59 +/- 1 years, p < 0.01) and more likely to have LVH (p < 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)