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Biomedical subjects

S M Hwang

Publications and source records attributed to S M Hwang.

At least 73 records · Page 4Linked to original sources

Differential absorption of amoxicillin from the human small and large intestine.

Differences in extent of amoxicillin absorption from various regions of the gastrointestinal tract were determined and compared with the same dose administered orally. Nine healthy men were intubated at a proximal (duodenum or jejunum) or distal (ileum or colon) site with use of a 15-foot double lumen nasointestinal tube. Amoxicillin solutions (375 mg in 120 ml water) were delivered on 2 successive days as a bolus or a 4-hour infusion. Subjects were reintubated at another site and amoxicillin administration was repeated. Subjects with colonic intubation received only infusions. Finally, all subjects received an oral dose of amoxicillin solution. Plasma samples were obtained at 16 time points over a 10-hour period and assayed for amoxicillin by use of an HPLC method. Area under the concentration-time curve and the maximum plasma concentration were computed to evaluate amoxicillin absorption. Amoxicillin absorption was rate and site dependent in the gastrointestinal tract. The drug was well absorbed in the duodenum and jejunum, with no significant differences in absorption when administered as a bolus or 4-hour infusion, but absorption was decreased and rate dependent in the ileum, where more drug was absorbed as an infusion compared with a bolus. Amoxicillin was unabsorbed when infused in all colonic regions.

Adult↗

Structural aspects of the reversal phase of alveolar bone remodelling.

The ultrastructure of the reversal line during alveolar bone remodelling was investigated in the rat. Surface bone remodelling along the periosteum of the mandible was induced by the extraction of opposing maxillary molars. With transmission EM the reversal line was seen to be composed of a superficial electron-dense amorphous layer and a deep filamentous layer at 7 d after extraction. The reversal line exhibited strong alkaline phosphatase activity and contained acid mucopolysaccharide. Scanning EM of the surface of the line, exposed by sonication in distilled water, showed papillary structures, the surface of which appeared granular and exhibited a crystalline appearance. The tips of collagen fibrils of new bone were attached to the top of the papillae in the front area of bone formation. It is suggested that the reversal line is involved in the coupling of bone resorption and formation.

Alkaline Phosphatase↗

Investigation of conformational specificity at GPIIb/IIIa: evaluation of conformationally constrained RGD peptides.

RGD-containing proteins and peptides are known to bind to the platelet GPIIb/IIIa receptor and inhibit platelet aggregation. That a conformational component to the specificity exists is suggested by significantly lower activity of linear RGD analogs relative to closely related cyclic peptides and small proteins containing the RGD sequence. Recently, conformations for a suite of RGD containing cyclic peptides have been defined by NMR-based methods and, for one molecule, by X-ray diffraction. We report here the NMR-based conformational analysis of an additional cyclic peptide, cyclo(Pro-Arg-Gly-Asp-D-Pro-Gly), and compare the conformational variations in the suite of peptides and related analogs. Biological activity data for these peptides shows a preference of the platelet GPIIb/IIIa receptor for one conformation of the RGD sequence, but suggests its ability to bind a second, distinct conformation.

Amino Acid Sequence↗

Stimulation of leukotriene production and membrane translocation of 5-lipoxygenase by cross-linking of the IgE receptors in RBL-2H3 cells.

Recent studies in rat basophilic leukemia cells (RBL-2H3) have shown that two pharmacological agents, ionomycin and thapsigargin, induce leukotriene C4 production and translocation of 5-lipoxygenase from cytosol to membrane, primarily by causing an influx of extracellular calcium. In the present study, we investigate the induction of these events by receptor activation. Cross-linking of high-affinity IgE receptors (Fc epsilon RI) by antigen in RBL-2H3 cells leads to leukotriene C4 production and membrane translocation of 5-lipoxygenase. As in the ionomycin-stimulated cells, leukotriene C4 production in antigen-stimulated cells is calcium-dependent since the amount of leukotriene C4 produced correlates quantitatively with the increase in intracellular free calcium concentration ([Ca2+]i). However, the increase in [Ca2+]i required for equivalent leukotriene C4 production by antigen is not as high as it is using ionomycin. In addition, no threshold [Ca2+]i level is required for leukotriene production by antigen, which is in contrast to the ionomycin stimulation that a [Ca2+]i level of 300-400 nM is required. Furthermore, antigen causes an additive increase in leukotriene C4 production in cells stimulated by the ionomycin. These results suggest that another as yet unidentified intracellular pathway acts in conjunction with Ca2+ for leukotriene synthesis in antigen-stimulated cells. Antigen stimulation causes 20-30% of the total cell 5-lipoxygenase to associate with membranes (compared with 10% in unstimulated cells) as demonstrated by enzyme activity assay and by Western Blot using antibodies to 5-lipoxygenase.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influx of extracellular calcium is required for the membrane translocation of 5-lipoxygenase and leukotriene synthesis.

Our studies assessed the effects of increases in intracellular calcium concentrations [( Ca2+]i) on leukotriene synthesis and membrane translocation of 5-lipoxygenase (5LO). The calcium ionophore ionomycin and the tumor promoter thapsigargin stimulated leukotriene production and translocation of 5-lipoxygenase to the membrane. Both agents elicited prolonged rises in [Ca2+]i. Leukotriene C4 production associated with [Ca2+]i in cells stimulated with various concentrations of ionomycin and thapsigargin suggests that a threshold [Ca2+]i level of approximately 300-400 nM is required. In the absence of extracellular Ca2+, both the ionomycin- and thapsigargin-induced rises in [Ca2+]i were transient, indicating that the prolonged [Ca2+]i elevation is due to an influx of extracellular Ca2+. Addition of EGTA to the external medium before, or at different times during, the treatment with ionomycin or thapsigargin instantaneously inhibited 5LO translocation and leukotriene synthesis, indicating that Ca2+ influx plays an essential role in 5LO membrane translocation and leukotriene synthesis. No leukotriene production was detected when cells were stimulated by a physiological stimulus of leukotriene D4. The addition of 100 nM leukotriene D4 triggered peak rises in [Ca2+]i that were comparable to those achieved by the ionomycin and thapsigargin. However, the leukotriene D4 induced rise was transient and rapidly declined to a lower but still elevated steady-state level, which was attributed to Ca2+ influx. Stimulation with 100 nM leukotriene D4 for 15 s increased the cellular levels of 1,4,5-inositol triphosphate (IP3), 1,3,4-IP3, and 1,3,4,5-inositol tetraphosphate (IP4).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Evaluating the safety and potential use of a weight-bearing exercise, Tai-Chi Chuan, for rheumatoid arthritis patients.

The safety of a traditional Chinese exercise, Tai-Chi Chuan, on rheumatoid arthritis (RA) patients was evaluated. RA patients, who received 1 h of Tai-Chi Chuan instruction once (n = 20) and twice (n = 15) a week for 10 consecutive wk in two separate studies, showed no deterioration in their clinical disease activities compared with the corresponding controls (n = 11 and 9, respectively). Testing parameters included joint tenderness, joint swelling, time to walk 50 feet, handgrip strength and a written functional assessment. No significant exacerbation of joint symptoms using this weight-bearing form of exercise was observed. Tai-Chi Chuan exercise appears to be safe for RA patients and may serve as an alternative for their exercise therapy and part of their rehabilitation program. Weight-bearing exercises have the potential advantages of stimulating bone growth and strengthening connective tissue, but this effect needs to be documented in long-term studies.

Adult↗

Folate deficiency and an abnormal lymphocyte deoxyuridine suppression test in monkeys.

Cebus albifrons were fed folate-deficient diets in order to assess folate status at the cellular level with the deoxyuridine suppression test. Plasma and red blood cell folates were significantly lower at 2 months, compared to control values. Hematologic signs of megaloblastic anemia occurred after 6 months, with significantly lower hematocrit, hemoglobin and red blood cell number values and increased polymorphonuclear leukocyte lobe counts. Urinary formiminoglutamic acid excretion also was elevated significantly. Whole blood lymphocyte cultures exhibited abnormal deoxyuridine suppression of [3H]-thymidine incorporation into DNA with folate deficiency. Thus this deoxyuridine suppression test can be used in isolated whole blood lymphocytes of these nonhuman primates to document folate deficiency.

Animals↗

[Lysine production by Brevibacterium divaricatum NTU-2 and its recovery from the fermentation broth].

An accumulation of L-lysine of about 42 g/l (as L-lysine-HCl) was obtained by cultivating Brevibacterium divaricatum NTU-2 in a medium containing 10.0% glucose, 4.0% (NH4)2 SO4, 0.1% KH2 PO4, 0.04% Mg SO4.7H2 O, 30 ml/l soybean meal hydrolysate, 50 mg/l DL-methionine, 100 micrograms/l d-biotin, 100 micrograms/l thiamine-HCl and 5% CaCO3 at pH 7.0. The yield was about 48.8% based on consumed glucose. The L-lysine accumulated in the broth was recovered and purified by simply using a strong cation-exchange resin, Diaion SK1B. The absorbed L-lysine was eluted with 4% ammonia water, and the average adsorption and desorption rates estimated (by 10 repeated experiments) were found to be 96.1% and 97.5%, respectively. The eluate was concentrated to about 50% by volume and adjusted to pH 4.0 with 8 N HCl. The solution was concentrated again to one half volume and followed by adding three volumes of 95% ethanol. The crude crystals of L-lysine-HCl in purity of 89.5% were obtained by filtration. The recovery was 93.5%. The resulting crude crystals were then dissolved in water, and subjected to decolorization with 0.5% active carbon. The decolorized solution thus obtained was recrystalized by concentration and 95% ethanol addition. After filtration and drying at 80 degrees C for 3 hr, white crystalline L-lysine with purity of 99.2% was finally obtained. The overall recovery of L-lysine was up to 86.7%.

Brevibacterium↗

Lymph node blast crisis of chronic myelogenous leukemia. One case report.

We present a patient with philadelphia chromosome (ph')-positive CML with extramedullary blast crisis in the lymph nodes before the bone marrow blast crisis. The morphological and cytochemical studies of lymph node touch preparation, and cytogenetic studies of lymph node tissue confirmed the extramedullary lymphoid blast transformation of CML, while the bone marrow continued to show the chronic phase of CML. When we changed our treatment to use Vincristine-Prednisolone therapy, the lymphadenopathy disappeared gradually. This case shows that the clinical and prognostic features of extramedullary disease can strongly affect disease acceleration in CML, and the therapy should be changed promptly. Besides, cytogenetic study of extramedullary tumor tissue in CML can be useful for accurate diagnosis.

Aged↗

A simple method for observation of phagocytosis on bacterial thin-layer.

A simple method was developed to visually present the phagocytic activity of leukocytes by using adherent Staphylococcus aureus cells and blood applied on a plastic dish. When heparinized blood was applied on thin-layer of heat-killed S. aureus cells on the plastic dish, plaques due to the phagocytic activity of leukocytes were observed with a microscope under a low magnification. Fewer and smaller plaques were observed when plasma-deprived rather than whole blood was used. Some analyses were made in respect to the fundamental conditions required for optimal results. This method was considered to be useful for conveniently evaluating the serum opsonin activities and phagocytic function of leukocytes in various kinds of diseases.

Antigens, Bacterial↗

Simple method for quantitation of cell-bound protein A on Staphylococcus aureus cells by means of hemagglutination with sheep erythrocytes differentially sensitized with rabbit antibody and its clinical application.

A simple method for quantitation of cell-bound protein A (SpA) on organisms of Staphylococcus aureus was successfully devised by using hemagglutination between staphylococcal organisms and a series of sheep erythrocyte suspensions sensitized with different amounts of anti sheep erythrocyte rabbit antiserum. The validity of the principle and the reproducibility of the method presented here were precisely analyzed and the details of the method are presented. The hemagglutination was quantitatively inhibited both by normal rabbit serum and by soluble SpA. Using the method presented here, 376 strains of S. aureus freshly isolated from clinical materials were subjected to SpA quantitation in cell-bound form. According to the results, there was an unexpected distribution profile in the amounts of cell-bound SpA among the clinical isolates, which showed a two-peak pattern. A possible usefulness of the method presented here in clinical investigations is briefly discussed also.

Animals↗

Improved methods for detection and serotyping of coagulase from Staphylococcus aureus.

Improved methods for detection and serotyping of staphylocoagulase were concomitantly devised. We devised an improved method for detection of coagulase activity on agarose film in the same manner as single radial immunodiffusion. The amounts of reagents required for detection of coagulase on agarose film were successfully diminished by adding polyethylene glycol (PEG) to the original formula described by Boothby et al. Using microplates in another improved method for coagulase serotyping, the amount of reaction fluid required was considerably less compared with the conventional tube method. PEG was found to be also effective to increase the efficacy of coagulase serotyping. In the presence or absence of anti-coagulase antisera, culture supernatants of staphylococcal strain grown in brain heart infusion broth were incubated with the reaction fluid containing bovine fibrinogen, rabbit plasma, 6-amino caproic acid, polyethylene glycol 6,000. Coagulase activity was visualized as a turbid mass formed in the wells. Turbid mass formation due to coagulase activity was type-specifically inhibited in the presence of type-specific antisera. Detailed procedures of the methods are precisely described with some preliminary results obtained by the methods.

Coagulase↗

Isolation and characterization of a cDNA clone encoding rat 5-lipoxygenase.

A full-length cDNA clone encoding 5-lipoxygenase, a key enzyme in the formation of leukotrienes, was isolated from a rat basophilic leukemia cell lambda gt11 cDNA library. The 2.5-kilobase (kb) cDNA insert, whose identity was confirmed by hybrid-select translation and DNA sequence analysis, has a 2.0-kb open reading frame encoding a protein of Mr approximately 77,600 and includes 60 base pairs of 5'-untranslated region and 0.4 kb of 3'-untranslated region to the polyadenylation signal. The deduced amino acid sequence shows significant homology with published sequences for the rabbit reticulocyte lipoxygenase and soybean lipoxygenase-1; it also contains sequences similar to a consensus sequence found in several calcium-dependent membrane-binding proteins. The cDNA recognizes a 2.6-kb mRNA species which is detected in all tissues but is particularly abundant in RNA from lung.

Amino Acid Sequence↗

Interactions of 5-lipoxygenase with membranes: studies on the association of soluble enzyme with membranes and alterations in enzyme activity.

Treatment of rat basophilic leukemia cells (RBL-1) with the calcium ionophore A23187 resulted in activation of 5-lipoxygenase, as indicated by an induction of leukotriene release [Orning, L., Hammarström, S., & Samuelsson, B. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 2017]. The enzyme activation was accompanied by a time-dependent association of 5-lipoxygenase to the particular fraction. When cells were lysed in the presence of 0.05-10 microM CaCl2, the soluble 5-lipoxygenase became associated with the particulate fraction. This was demonstrated by a decrease in immunoreactivities and enzymatic activities in the soluble fraction and a parallel increase in particulate-associated immunoreactivities. The particulate-bound enzyme was not active. Ca2+ induced the membrane association of 5-lipoxygenase when added into the incubation mixtures containing the membrane fraction with either the cytosolic fraction or the purified enzyme. 5-Lipoxygenase also bound to the microsomal-enriched fraction in the presence of Ca2+. Maximal membrane binding was obtained after a 1-min incubation at 4 degrees C. When a fixed amount of isolated membranes (0.2 mg of protein) and increasing cytosolic protein (0.5-4 mg) were used, a linear increase in enzyme binding was observed. The binding became saturated at 3 mg of cytosolic protein/mg of membrane protein. 5-Lipoxygenase binding to the membrane fraction was unaffected by pretreatment of the membranes with trypsin but was inhibited by treating with phospholipase A2, suggesting that phospholipids are involved.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Structure-activity relationships of beta-adrenergic receptor-coupled adenylate cyclase: implications of a redox mechanism for the action of agonists at beta-adrenergic receptors.

The present studies have tested the hypothesis that agonists at beta-adrenergic receptors activate the beta-receptors by reducing them. This was examined by analyzing the interactions of 41 beta-agonists and antagonists with the receptors. The structural features which determined binding affinity (KD) were shown to be distinct from those which determined intrinsic, activity (IA). The IA was shown to be related to the oxidation-reduction properties which were determined primarily by the nature of the substituents on the phenyl ring. Thus, the parent compound phenylethanolamine, having no phenolic substituent, acted as an antagonist (IA = 0) and was also redox inactive. All of the antagonists tested (19) exhibited EP (peak potential for the first oxidative wave) values greater than 0.75 V, suggesting that they were difficult to oxidize. Agonists, however, exhibited a wide range of EP (0.25-0.7 V) with values lower than those of the antagonists. The agonists tested include catecholamines, catecholamine analogs bearing meta-substituted amino functionalities (such as amino, methylamino, formanilide, sulfonamide, urea, and carbamate), resorcinol, and hydroxymethyl congeners. It is proposed that the oxidizing tendency of the substituent on the phenyl ring is one of the factors that influences IA. To test the hypothesis further, we electrolytically oxidized isoproterenol to adrenochrome or to the o-quinone intermediate and tested for activity. The 4e-, 4H+-oxidation product adrenochrome did not bind to or stimulate adenylate cyclase, suggesting that the reducing ability to isoproterenol is important for its agonistic activity. A cyclic redox mechanism for the action of agonists at beta-adrenergic receptors is presented. We propose that agonist are electron donors. Their interactions with receptors result in reduction leading to activation of the receptors.

Adenylyl Cyclases↗

Characteristics of lysine uptake by isolated renal cortical tubule fragments from mature and immature dogs.

The uptake of L-lysine was examined in isolated renal cortical tubule fragments from adult and 1-week-old dogs. Lysine uptake by adult tubules was initially more rapid than that by the immature tubules. This uptake by mature tubules reached a steady state after 30 min of incubation, while the newborn tubules still had not reached a steady state by 90 min of incubation. Because a steady state of lysine uptake was not attained with the immature tubules, their uptake of lysine exceeded that of the adult after 60 min of incubation. Kinetic studies revealed that lysine was taken up by one saturable transport system with a Km of 0.56 mM and Vmax of 6.18 mmol/liter intercellular fluid per 5 min in the adult and one saturable transport system in the 1-week-old with a Km of 0.38 mM and Vmax of 3.66 mmol/l intracellular fluid per 5 min. Lysine also entered the renal tubule cells in both age groups via a diffusional pathway with a kd of 0.35 min-1 in the adult and 0.30 min-1 in the newborn. Cystine competitively inhibited lysine uptake by adult dog tubules with a Ki of 0.61 mM. The other dibasic amino acids, ornithine and arginine, also inhibited lysine uptake in both the adult and the newborn.

Age Factors↗