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Biomedical subjects

H S Ju

Publications and source records attributed to H S Ju.

9 recordsLinked to original sources

Characterization of an extracellular poly(3-hydroxy-5-phenylvalerate) depolymerase from Xanthomonas sp. JS02.

A bacterium, JS02, capable of degrading an aromatic medium-chain-length polyhydroxyalkanoate (PHA(MCL)), poly(3-hydroxy-5-phenylvalerate) (PHPV), was isolated from wastewater-treatment sludge (Ju et al. 1998), and was identified as a Xanthomonas species. An extracellular PHPV depolymerase was purified from the concentrated culture broth of Xanthomonas sp. JS02 by using a chromatography series on Sephadex G-75, QAE-Sephadex A-50 and hydroxyapatite. The molecular mass of the purified enzyme was estimated to be 41.7 kDa. The purified enzyme could hydrolyse PHPV and p-nitrophenyl (PNP)-esters of fatty acids, but did not hydrolyse short-chain-length PHAs, though the culture supernatant could hydrolyse them. The optimum pH range was 8.0-9.0 and the optimum temperature was 60 degrees C for PNP-octanoate hydrolysis. The Km values for PNP-hexanoate and PNP-octanoate were 10.9 and 0.88 microM, respectively.

Biodegradation, Environmental↗

Development of an immunoassay for urinary galactosylhydroxylysine.

Galactosylhydroxylysine (GHL) is released during bone resorption and has been shown to be elevated in subjects with metabolic bone loss. GHL is relatively specific for bone, it is not recycled or significantly metabolized during collagen turnover, and the levels are not influenced by diet. Previous measurements of GHL levels in urine have been performed using reverse-phase high performance liquid chromatography following pre-column derivatization. We produced polyclonal antibodies to GHL using GHL purified from sea sponges and developed an immunoassay that can recognize GHL in urine. The antibodies have minimal cross-reactivity with a physiological mixture of amino acids (< 1%), galactose (< 0.2%), lactose (< 0.3%), and glucosylgalactosylhydroxylysine (< 1%). This competitive immunoassay requires no dilution or pretreatment of the samples and provides a rapid and easy method for the evaluation of GHL in urine. Analysis of clinical samples from normal individuals, post-menopausal women, osteoporotic patients and individuals with Paget's disease show that the assay can discriminate between groups with differing levels of bone resorption as well as deoxypyridinoline (Dpd).

Adult↗

Comparison of analytical performance and biological variability of three bone resorption assays.

We have compared the analytical performance and biological variability of three commercially available bone resorption assays: Pyrilinks-D, Osteomark, and CrossLaps, for the measurement of urinary free deoxypyridinoline (Dpd), cross-linked N-telopeptides of type I collagen (NTx), and linear C-telopeptides of type I collagen (CTx), respectively. The intraassay and interassay CVs for precision of the Dpd and NTx assays were < 10% for analyte concentrations greater than the second calibrator (i.e., 3 nmol/L Dpd or 30 nmol bone collagen equivalents/L NTx). The CTx assay demonstrated poor precision for analyte concentration lower than the third calibrator (i.e., 200 micrograms/L). The NTx assay exhibited nonlinear recovery for sample dilutions prepared in buffer; however, this nonlinear recovery could be corrected for sample dilutions made in urine at a low analyte concentration. Supplement recoveries of each of the three assays were within 100% +/- 10% on average. All three analytes showed stability through five freeze-thaw cycles. The mean day-to-day variations were 16% for Dpd, and 23% for both NTx and CTx. Similar diurnal rhythm was observed for all three assays on average, with the peak in the early morning and the nadir in the afternoon. Mean amplitude of the diurnal variation was 37% for Dpd and NTx, and 57% for CTx. Variations within the reference intervals for a healthy premenopausal population were 28% for Dpd, 57% for NTx, and 56% for CTx. Pyrilinks-D has demonstrated analytical precision and accuracy equal or superior to Osteomark and CrossLaps in all areas. Dpd exhibits the least biological variability day-to-day, within individuals across the diurnal cycle, and within a healthy premenopausal population.

Adult↗

Glycerol-3-phosphate dehydrogenase activity in human lymphocytes: effects of insulin, obesity and weight loss.

Insulin exposure stimulates an increase in glycerol-3-phosphate dehydrogenase (G3PDH) activity in isolated human lymphocytes that correlates to an increase in G3PDH mRNA and requires new protein synthesis. Synthetic diacylglycerol or phorbol ester can mimic the effect of insulin on G3PDH activity, suggesting that protein kinase C may be involved in regulation of G3PDH levels. In addition, lithium chloride, an inositol phosphate phosphatase inhibitor, and calcium uptake inhibitors can abolish insulin stimulation of G3PDH activity. For obese subjects in whom insulin resistance in vitro can be demonstrated, the extent of insulin stimulation of G3PDH activity is decreased compared to normal weight individuals, and treatment by a very low calorie diet restores insulin stimulation of G3PDH activity. Thus, insulin stimulation of G3PDH activity is dependent upon the metabolic state of the subject from whom the cells are obtained.

Diet, Reducing↗

Mechanisms of protective effects of berbamine on ischemia/reperfusion injury in isolated rat heart.

Isolated perfused rat heart model was used to observe the protective effects of berbamine on myocardial ischemia/reperfusion injury. The hearts were significantly injured by 40 min global ischemia followed by 20 min reperfusion. Berbamine could significantly improve heart function, prevent ventricular fibrillation, reduce CK release, preserve Na,K-ATPase activity, and reduce Na+ gain and K+ loss during ischemia and Ca2+ overload during reperfusion. With the use of low temperature ESR technique, in hearts subjected to 40 min ischemia and 15 sec reperfusion, oxygen-centered free radical signals became much more intense. In the presence of berbamine, these signals decreased. Results showed that berbamine could alleviate myocardial ischemia/reperfusion injury. This effect might be due to: 1) preserved myocardial Na,K-ATPase activity and inhibition of sodium overload at the end of ischemia, which might further lead to attenuation of reperfusion-induced calcium overload, and 2) reduction of oxygen free radical generation during reperfusion.

Alkaloids↗

Scavenging effect of berbamine on active oxygen radicals in phorbol ester-stimulated human polymorphonuclear leukocytes.

The scavenging effect of berbamine (Ber) on active oxygen radicals was studied, using a spin-trapping technique and a chemiluminescence (CL) method in phorbol myristate acetate (PMA) stimulated polymorphonuclear leukocytes (PMN) and in four cell-free superoxide (O2-.) or hydroxyl radical (OH.) generating systems. Ber (0.1 to 0.3 mM) effectively reduced active oxygen radicals in PMN stimulated with PMA, but had no obvious effect on oxygen consumption during the respiratory burst of PMN, measured with spin probe oxymetry. Ber (0.3 mM) prominently inhibited the CL response of PMA-stimulated PMN. The agent remarkably quenched O2-. in xanthine/xanthine oxidase and irradiation riboflavin systems and OH. in the Fenton reaction. Its scavenging action on O2-. was stronger than that of Vitamin E in the xanthine/xanthine oxidase system but the same as Vitamin E in the riboflavin system, and its action on OH. was similar to that of Vitamin E.

Alkaloids↗

[Scavenging effects of sodium ferulate and 18 beta-glycyrrhetic acid on oxygen free radicals].

The scavenging effects of sodium ferulate (SF) and glycyrrhetic acid (GA) on oxygen free radicals were studied by using spin trapping technique and chemiluminescence (CL) methods. SF 0.3 mmol/L and GA 0.3 mmol/L effectively scavenged oxygen free radicals produced by phorbol myristate acetate (PMA)-stimulated polymorphonuclear leukocytes (PMN), but had no effect on the oxygen consumption by PMN, measured with spin probe oxymetry. In 3 cell-free oxygen free radical generating systems, both SF and GA showed a remarkably scavenging effects on O2- and OH.. SF 30 mumol/L and GA 30 mumol/L prominently inhibited the CL response of PMA-stimulated PMN. The results demonstrated that SF and GA are oxygen free radical scavengers.

Coumaric Acids↗

[Antioxidant effect of berbamine].

The antiperoxidant action of berbamine (Ber) was demonstrated by colorimetric estimation of malondialdehyde (MDA) formation, and the scavenging effect of Ber on O2- was investigated by chemiluminescence method and ESR-spin trapping technique. Ber 11-100 mumol/L remarkably inhibited MDA formation induced by incubating mice liver homogenate at 37 degrees C with vibration for 1 h. Ber 1-100 mumol/L and 0.2-0.6 mmol/L effectively scavenged O2- in alkaline DMSO and xanthine/xanthine oxidase systems respectively. The results show that Ber is an antioxidant.

Alkaloids↗

[Effects of glycyrrhiza flavonoid on lipid peroxidation and active oxygen radicals].

The antiperoxidant activity of glycyrrhiza flavonoid (FG) was studied by using colorimetric estimation of lipid peroxide (MDA) formation. The scavenging effects of FG on O2-. and OH. was investigated by using chemiluminescence method and spin trapping technique in different systems. The results were as follows: FG 2.8-25 micrograms/ml effectively inhibited MDA formation induced by incubating mice liver homogenate at 37 degrees C for 1 h; FG 0.265-26.5 micrograms/ml or 2.58-258 micrograms/ml was shown to markedly scavenge O2-. in alkaline/DMSO or xanthine/xanthine oxidase systems, respectively in a concentration-dependent manner. FG 144 micrograms/ml or 258 micrograms/ml also significantly scavenged the active oxygen radicals produced by PMN stimulation with PMA or OH. produced in Fenton's reaction respectively. The results suggest that FG may be used as an antioxidant.

Animals↗