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Biomedical subjects

Y H Kang

Publications and source records attributed to Y H Kang.

At least 37 records · Page 2Linked to original sources

Lignans from Kadsura angustifolia.

A new dibenzocyclooctadiene lignan named angustifolin D (1) together with four known lignans: kadsulignan L (2), kadsulignan N (3), schisantherin P (4) and meso-dihydroguaiaretic acid (5) were isolated from the stems of Kadsura angustifolia. Their structures and stereochemistries were elucidated by spectral studies. Compounds 2 and 5 showed moderate platelet-activating factor (PAF) antagonistic activities with IC50 values of 2.6 x 10(-5) and 4.1 x 10(-5) M, respectively.

Animals↗

Truncated SNAP-25 (1-197), like botulinum neurotoxin A, can inhibit insulin secretion from HIT-T15 insulinoma cells.

We and others have previously shown that insulin-secreting cells of the pancreas express high levels of SNAP-25 (synaptosomal-associated protein of 25 kDa), a 206-amino acid t-SNARE (target soluble N-ethylmaleimide-sensitive factor attachment protein receptors) implicated in synaptic vesicle exocytosis. In the present study, we show that SNAP-25 is required for insulin secretion by transient transfection of Botulinum Neurotoxin A (BoNT/A) into insulin-secreting HIT-T15 cells. Transient expression of BoNT/A cleaved the endogenous as well as overexpressed SNAP-25 proteins and caused significant reductions in K+ and glucose-evoked secretion of insulin. To determine whether the inhibition of release was due to the depletion of functional SNAP-25 or the accumulation of proteolytic by-products, we transfected cells with SNAP-25 proteins from which the C-terminal nine amino acids had been deleted to mimic the effects of the toxin. This modified SNAP-25 (amino acids 1-197) remained bound to the plasma membrane but was as effective as the toxin at inhibiting insulin secretion. Microfluorimetry revealed that the inhibition of secretion was due neither to changes in basal cytosolic Ca2+ levels nor in Ca2+ influx evoked by K(+)-mediated plasma membrane depolarization. Electron microscopy revealed that cells transfected with either BoNT/A or truncated SNAP-25 contained significantly higher numbers of insulin granules, many of which clustered close to the plasma membrane. Together, these results demonstrate that functional SNAP-25 proteins are required for insulin secretion and suggest that the inhibitory action of BoNT/A toxin on insulin secretion is in part caused by the production of the plasma membrane-bound cleavage product, which itself interferes with insulin granule docking and fusion.

Botulinum Toxins, Type A↗

Phacoemulsification and posterior chamber intraocular lens implantation after scleral buckling, vitrectomy, or both.

BACKGROUND AND OBJECTIVE: Previous studies have documented the risk of complications for eyes with prior vitreoretinal surgery that later undergo intracapsular or extracapsular cataract extraction. The authors reviewed the records of patients with a history of vitreoretinal surgery and subsequent phacoemulsification to assess the visual outcome in the relatively early postoperative period and to determine the risk of postoperative complications. PATIENTS AND METHODS: The authors identified 21 eyes of 21 patients with a history of scleral buckling, vitrectomy, or both, who subsequently underwent phacoemulsification. Data recorded before, during, and after surgery were reviewed. RESULTS: The best-corrected visual acuity improved in 20 (95%) of 21 eyes, and the final best-corrected visual acuity was 20/40 or better in 12 (57%) of 21 eyes. The intraoperative complications included iridodialysis, zonular dialysis, and posterior capsular rupture. The immediate postoperative complications were microcorneal edema, anterior chamber hyphema, vitreous hemorrhage, and fibrinous pupillary membrane. No retinal detachments occurred during the postoperative follow-up period, which ranged from 3 to 10 months. Late complications included isolated cases of posterior capsular opacity, herpetic corneal ulcer, and iris posterior synechia; however, the final visual acuities were not limited, except in one eye with a corneal ulcer. CONCLUSION: Although some postoperative complications did occur, they did not limit final vision, and useful vision was achieved.

Adolescent↗

Cutaneous lesions in swine after decompression: histopathology and ultrastructure.

A detailed histopathologic description of skin lesions from a porcine model of decompression sickness (DCS) is presented. Pigs were dived in a dry chamber on a variety of profiles over an 11-mo period, with a 0.1-0.6 (10-60%) incidence of cutaneous lesions. The clinical appearance of the lesions evolved from irregular, sharply demarcated areas of erythema to violaceous and, eventually, darkly mottled macules. The lesions were biopsied under deep, sedative anesthesia. Histologic abnormalities were found in 91% (20/22) of the biopsies from clinically apparent cutaneous lesions. Vascular congestion was the most common finding. Focal areas of vasculitis were noted in 45% (10/22) of the lesions. Perivascular neutrophil infiltrates, edema, and occasionally, hemorrhage were also noted. Ultrastructural abnormalities were found in all of the lesions studied. Acute inflammation affecting the dermal vasculature was the most common finding. Platelets were rarely observed aggregating within vessels. The clinical and histologic features of cutaneous lesions in pigs after decompression are compared with previous accounts in humans. The model provides a useful tool for the study of cutaneous lesions in DCS and may be a means of exploring interventions in the disease.

Animals↗

New generation dopaminergic agents. 1. Discovery of a novel scaffold which embraces the D2 agonist pharmacophore. Structure-activity relationships of a series of 2-(aminomethyl)chromans.

A series of 2-(aminomethyl)chromans (2-AMCs) was synthesized and evaluated for their affinity and selectivity for both the high- and low-affinity agonist states (D2High and D2Low, respectively) of the dopamine (DA) D2 receptor. The 7-hydroxy-2-(aminomethyl)chroman moiety was observed to be the primary D2 agonist pharmacophore. The 2-methylchroman moiety was discovered to be an entirely novel scaffold which could be used to access the D2 agonist pharmacophore. Attaching various simple alkyl and arylalkyl side chains to the 7-hydroxy 2-AMC nucleus had significant effects on selectivity for the D2High receptor vs the 5HT1A and alpha 1 receptors. A novel DA partial agonist, (R)-(-)-2-(benzylamino)methyl)chroman-7-ol [R-(-)-35c], was identified as having the highest affinity and best selectivity for the D2High receptor vs the alpha 1 and 5HT1A receptors. Several regions of the 2-AMC nucleus were modified and recognized as potential sites to modulate the level of intrinsic activity. The global minimum conformer of the 7-hydroxy-2-AMC moiety was identified as fulfilling the McDermed model D2 agonist pharmacophoric criteria and was proposed as the D2 receptor-bound conformation. Structure-activity relationships gained from these studies have aided in the synthesis of D2 partial agonists of varying intrinsic activity levels. These agents should be of therapeutic value in treating disorders resulting from hypo- and hyperdopaminergic activity, without the side effects associated with complete D2 agonism or antagonism.

Animals↗

Autologous lymphoblastoid cell lines stably transfected with Plasmodium falciparum circumsporozoite protein as targets in cytotoxic T-lymphocyte assays.

To produce cell lines that can be used as a continuous source of antigen presenting cells for stimulating T-cell lines and clones and as targets in cytotoxic T-lymphocyte (CTL) assays, we used a retroviral vector with a simian virus (SV40) early promotor to transfer a Plasmodium falciparum circumporozoite (PfCSP) gene into human EBV transformed B-lymphoblastoid cell lines (B-LCL). We herein report successful, stable transfection and cell surface expression of this gene, as confirmed by PCR, Western blot analysis and immunoelectron microscopy. One of three successfully transfected autologous cell lines expressed PfCSP on the cell surface and was lysed by CD8+ T-cell dependent CTL from a donor volunteer who had been immunized with irradiated P. falciparum sporozoites. Such cell lines should provide excellent tools for characterizing human CD8+ T-cell responses against Plasmodium sp. proteins.

Animals↗

Metabolic cleavage of frangufoline in rodents: in vitro and in vivo study.

Frangufoline, a sedative 14-membered frangulanine-type cyclopeptide alkaloid, was found to be rapidly converted, via enzymatic process, in vitro and in vivo in rodents to M1 ((S)-(N,N-dimethylphenylalanyl)-(2S,3S)-3-[(p-formylphenoxy) leucyl]-(S)-leucine); which is a substituted linear tripeptide. The reaction did not require low molecular weight cofactors, and mammalian serum failed to catalyze the reaction. Structure-reactivity study of cyclopeptide alkaloid analogs suggested that the enamide bond is the site being cleaved, and the reaction was inhibited by organophosphorus esters such as BPNP and by eserine at higher concentrations but not by eserine at lower concentrations or by EDTA and PCMB. On the basis of these results, a possible mechanism for metabolic conversion of frangufoline to M1 was proposed, in which oxidation of the vinyl group and enzyme-catalyzed hydrolysis of the adjacent amide bond, possibly by B-esterase-like enzyme, proceed in a concerted manner.

Animals↗

Distribution of the cellular uptake of phosphorothioate oligodeoxynucleotides in the rat kidney in vivo.

Previous animal studies have demonstrated that following systemic administration phosphorothioate oligodeoxynucleotides (S-ODNs) are primarily excreted by the kidneys and that renal tissue levels of S-ODNs exceed that of other organs. Thus, the kidney may be an ideal target organ for application of antisense S-ODNs in vivo. We examined which cells within the rat kidney have uptake of radiolabeled S-ODNs following intravenous infusion. A 20-base 35S-ODN was infused into 6 adult male Wistar rats. Three animals each were sacrificed 30 min and 4 h after infusion. The kidneys were then removed, fixed, and tissue autoradiography was performed. Similar results were obtained in both groups. The highest level of radioactivity was seen within the proximal tubules. Lower levels of activity were seen within the glomerulus, the parietal epithelial cells of Bowman's space, and distal tubular cells. Very weak activity was also detected within the cells of the loop of Henle and the medullary collecting ducts. These results demonstrated that within the kidney S-ODNs were taken up primarily by proximal tubular cells, with much lower uptake by cells in other segments of the nephron.

Animals↗

Phacoemulsification and posterior chamber intraocular lens implantation in uveitis.

We reviewed the results of posterior chamber intraocular lens (PCL) implantation following scleral tunnel incision and phacoemulsification (PE) in 28 eyes with uveitis. Best-corrected visual acuity (BCVA) improved in 89% of eyes and the final BCVA was 20/40 or better in 64% of eyes. Intraoperative complication was a posterior capsular break (3 eyes). Postoperative complications were microcorneal edema (4 eyes), anterior chamber hyphema (3 eyes), posterior capsular opacity (3 eyes), fibrinous pupillary membrane (2 eyes), iris posterior synechia (2 eyes), neovascular glaucoma (1 eye), and retinal detachment (1 eye). In two eyes of three aftercataracts, visual improvement was achieved by Nd: YAG capsulotomy. A well-planned PE and PCL implantation through scleral tunnel in uveitis was not much more dangerous than expected.

Adolescent↗

Salicylate levels in rat stomach tissues after administration of aspalatone and acetylsalicylic acid in relation to their ulcerogenicity.

To study the mechanism for the low ulcerogenicity of the antithrombotic agent aspalatone ([3-[2-methyl-4-pyronyl)]-2-acetyloxybenzoate, CAS 147249-33-0), the metabolism and disposition of aspalatone were compared with those of acetylsalicylic acid (ASA) in the gut wall in relation to the salicylate level in the stomach tissues following oral administration in pyrolus-ligated rats. Both aspalatone and ASA were essentially stable in gastric juice and were absorbed in stomach unchanged. In glandular portion of the stomach, salicylate level found at 10 min post-dose in aspalatone (80 mg/kg)-and in ASA (50 mg/kg)-treated group was 67 +/- 43 nmol/g tissue and 2000 +/- 250 nmol/g tissue, respectively. In non-glandular (rumen) tissue, salicylate was not detected in the aspalatone group, whereas it reached a concentration of up to 1100 +/- 130 nmol/g tissue in the ASA group. As a result of the relative stability of the ester bond connecting the salicylic acid and maltol groups towards hydrolysis in the stomach and entrapment of ASA due to ion trapping, a lower salicylate level was observed in the stomach after oral aspalatone administration, and this may, at least in part, be the underlying mechanism for the low ulcerogenicity of aspalatone.

Animals↗

Reinforcement of PMMA bone cement with a continuous wire coil--a canine femur study.

The hoop and axial strains present due to loading after femoral stem implantation in canine femurs, implanted with and without a wire coil surrounding the distal tip were investigated. One stem served as the control without wire coil while the other was experimental reinforced with a wire coil. Both stems were implanted by identical methods. Ideally, the wire coil should serve to reduce the hoop and axial strains present in the distal tip of the arthroplasty. The strains were measured using 90 degrees rosette strain gages. Though the coil's position was altered slightly during implantation usable results were still obtained. At a maximum load of 44.5 N there were 32 and 19% reduction in the hoop and axial strains for the reinforced stem and the control, respectively. This experiment presents a striking difference between the control and reinforced hip arthroplasties. Equally important is that this study confirms the trends in hoop and axial strain behavior demonstrated in other works utilizing a wire coil reinforcement scheme. The simple method of applying a continuous wire coil may help to reduce the loosening of femoral stem of total hip arthroplasty by reducing strains at the tip of the stem due to the strengthening of the cement mantle.

Animals↗

Improvements in visual acuity following limbal lensectomy for subluxated lenses in children.

BACKGROUND AND OBJECTIVE: To report the surgical results of limbal lensectomy using an automated vitrectomy instrument for the treatment of subluxated lenses in children. PATIENTS AND METHODS: The authors reviewed the charts of children who underwent surgery for subluxated lenses. Limbal lensectomies with an "in the bag" technique were performed on 18 eyes of 10 patients. The indications for surgery were a best-corrected visual acuity of less than 20/60, bisection of pupil by the lens, or both. RESULTS: Best-corrected visual acuity improved in all 18 eyes (paired t test, P < .01). In 1 eye, transient anterior chamber hyphema occurred during the early postoperative period. No other significant complications were found during an average follow-up of 19.6 months. CONCLUSION: Limbal lensectomy for subluxated lenses in children improved visual acuity without severe complications.

Adolescent↗

Magnesium activated adenosine formation in intact perfused heart: predominance of ecto 5'-nucleotidase during hypermagnesemia.

Magnesium ion is an allosteric effector of 5'-nucleotidase and thus activates adenosine production from AMP. Two distinct 5'-nucleotidase systems, the membrane-bound ecto and the soluble cytosolic isoforms, exist in mammalian myocardium. The aim of this study was to delineate the contributions of the ecto vs. cytosolic isoforms to Mg2+-stimulated cardiac purine nucleoside formation and release. Isolated guinea pig hearts were retrogradely perfused at their physiological aortic pressure with Krebs-Henseleit bicarbonate buffer fortified with 10 mM glucose. AMP and the adenylate degradatives adenosine and inosine were measured in coronary venous effluent and in epicardial transudate, which was sampled to estimate concentrations of adenylate degradatives in the interstitium. When perfusate Mg2+ was increased from 0.6 to 6 mM, coronary vascular resistance and spontaneous heart rate fell, and steady-state coronary venous release of adenosine + inosine rose severalfold. Cytosolic free magnesium, as estimated by 31P-NMR after 15 min of perfusion with 6 mM Mg2+ or from chemically measured indicator metabolites after 30 min, rose 60 and 144% respectively (P < 0.05). Excess Mg2+ stimulated purine nucleoside release nearly threefold in coronary venous effluent and four- to sevenfold in epicardial transudate. 50 microM, alpha,beta-methylene adenosine 5'-diphosphate (AOPCP), a selective inhibitor of ecto 5'-nucleotidase, elevated interstitial AMP concentration tenfold, did not attenuate basal nucleoside release, but completely inhibited Mg2+-stimulated coronary venous purine nucleoside release and blunted Mg2+-stimulated interstitial purine nucleoside formation by 69%. During perfusion with exogenous 1 microM [8-14C]AMP, excess perfusate MgCl2 increased [14C]adenosine release by 63% in coronary effluent and 133% in epicardial transudate. AOPCP decreased baseline [14C]adenosine release in coronary effluent and epicardial transudate by 85-90%, caused equilibration of arterial and epicardial AMP, and attenuated MgCl2 activation of p[14C]adenosine formation by approx. 75%, in both the vascular and interstitial compartments. Intramyocytic concentrations of allosteric regulators of the cytosolic 5'-nucleotidases were evaluated in stop-frozen myocardium. Excess magnesium did not appreciably alter intracellular pH and ATP concentration, but lowered free cytosolic ADP and AMP concentrations by 50 and 70%, respectively. A simplified model of compartmentalized adenosine metabolism is proposed in which magnesium ion-activated cardiac purine release originates predominantly from the ecto 5'-nucleotidase; magnesium ion stimulation of metabolic flux through the cytosolic isoforms was constrained by concomitant reductions in intracellular AMP substrate and allosteric activator ADP. Magnesium ion-enhanced adenosine formation by 5'-nucleotidase could contribute to the known cardioprotective effects of this clinically used cation.

5'-Nucleotidase↗

Differential effects of transforming growth factor-beta 1 on lipopolysaccharide induction of endothelial adhesion molecules.

In this report, we studied the effects of transforming growth factor (TGF)-beta 1 on lipopolysaccharide (LPS)-induced expression of endothelial cell (EC) adhesion molecules. Confluent human umbilical cord vein EC cultures were stimulated with Escherichia coli LPS and TGF-beta 1, alone or in combination for various times and evaluated for expression of ICAM-1, E-selectin, and VCAM-1 by immunofluorescence and radioimmunoassay. Effects of LPS and/or TGF-beta 1 on cell growth were also studied by 3H-thymidine incorporation. Both LPS and TGF-beta 1 alone stimulated EC expression of the adhesion molecules in a dose-dependent manner. The effects of TGF-beta 1 on LPS induction of the adhesion molecules varied with LPS concentration and treatment time, mode, and duration. Pretreatment with TGF-beta 1 for 24 h greatly augmented LPS induction of ICAM-1 and VCAM-1 expression, but decreased E-selectin expression. TGF-beta 1 also enhanced expression of the adhesion molecules in cells that were pretreated with 1 microgram/mL LPS for 60 min. Concomitant treatment with TGF-beta 1/LPS resulted in significant increases in ICAM-1 but decreases in VCAM-1 expression. TGF-beta 1 effects on LPS induction of the adhesion molecules were more prominent at lower LPS levels (.001, .01 microgram/mL). Both LPS and TGF-beta 1 suppressed thymidine incorporation in a dose-related pattern. These data suggest that TGF-beta 1 has additive and antagonistic effects on LPS induction of the adhesion molecules and that the cell responsiveness to the stimuli in the expression is related to growth condition of the cells. In conclusion, our findings suggest that TGF-beta 1 exhibits pro-inflammatory and anti-inflammatory activities in human endothelial cells and may play an important role in regulating leukocyte adherence and extravasation under LPS-induced inflammatory conditions.

Cells, Cultured↗

Improved bonding strength of polyethylene/polymethylmetacrylate bone cement--a preliminary study.

The adhesion of polymethylmetacrylate (PMMA) bone cement and low density polyethylene (LDPE) substrate by means of corona discharge treatment are presented. The surfaces were characterized by water contact angle goniometer, Fourier-transform infrared spectroscopy in the attenuated total reflectance mode (FTIR-ATR), electron spectroscopy for chemical analysis (ESCA), and scanning electron microscopy (SEM) for surface morphology. The bonding strength between bone cement and LDPE with varying intensity of corona treatment was evaluated by a 180 degrees peel test. The water contact angles decreased with increasing treatment intensity from 95.0 +/- 1.8 to around 43.5 degrees. It was observed that the oxygen containing groups as hydroxyl, ether, carboxylic acid, ester, ketone or aldehyde groups were introduced on the LDPE surfaces by corona treatment. The peel strength steeply increased from 0 to 1.75 kgf/cm with increasing the intensity of corona treatment. The improved adhesion of bone cement onto LDPE substrate may be due to only secondary force such as hydrogen bonding between PMMA bone cement and hydrophilized LDPE substrate.

Adhesiveness↗

Mousepox outbreak in a laboratory mouse colony.

Mousepox was diagnosed in and eradicated from a laboratory mouse colony at the Naval Medical Research Institute. The outbreak began with increased mortality in a single room; subsequently, small numbers of animals in separate cages in other rooms were involved. Signs of disease were often mild, and overall mortality was low; BALB/cByJ mice were more severely affected, and many of them died spontaneously. Conjunctivitis was the most common clinical sign of disease in addition to occasional small, crusty scabs on sparsely haired or hairless areas of skin. Necropsy findings included conjunctivitis, enlarged spleen, and pale liver. Hemorrhage into the pyloric region of the stomach and proximal portion of the small intestine was observed in experimentally infected animals. In immune competent and immune deficient mice, the most common histologic finding was multifocal to coalescing splenic necrosis; necrosis was seen less frequently in liver, lymph nodes, and Peyer's patches. Necrosis was rarely observed in ovary, vagina, uterus, colon, or lung. Splenic necrosis often involved over 50% of the examined tissue, including white and red pulp. Hepatic necrosis was evident as either large, well-demarcated areas of coagulative necrosis or as multiple, random, interlacing bands of necrosis. Intracytoplasmic eosinophilic inclusion bodies were seen in conjunctival mucosae and haired palpebra. Ectromelia virus was confirmed as the causative agent of the epizootic by electron microscopy, immunohistochemistry, animal inoculations, serologic testing, virus isolation, and polymerase chain reaction. Serologic testing was of little value in the initial stages of the outbreak, although 6 weeks later, orthopoxvirus-specific antibody was detected in colony mice by indirect fluorescent antibody and enzyme-linked immunosorbent assay procedures. The outbreak originated from injection of mice with a contaminated, commercially produced, pooled mouse serum. The most relevant concern may be the unknown location of the source of the virus and the presence of a reservoir for this virus within the United States.

Animals↗

Effects of endotoxin on expression of VLA integrins by human bronchoalveolar lavage macrophages.

Endotoxin (lipopolysaccharide, LPS) is known to induce inflammatory responses, such as monocyte/macrophage adherence, migration, and accumulation. Recruitment and accumulation of macrophages during infection and inflammation are regulated by integrin-mediated cell-extracellular matrix interactions. In the present report, we studied the effects of LPS on the expression of VLA-5 (alpha 5 beta 1), VLA-3 (alpha 3 beta 1), and VLA-2 (alpha 2 beta 1) integrins and fibronectin (FN) by human alveolar macrophages in an attempt to understand the mechanism by which LPS regulates macrophage adhesion to matrix proteins. Bronchoalveolar lavage macrophages were treated with varying concentrations of Escherichia coli LPS for different times and evaluated for expression of the integrins and FN by immunofluorescence, immunoelectron microscopy, autoradiography, and radioimmunoassay. Immunofluorescent and immunoelectron microscopic observations showed that VLA integrins were constitutively expressed on the cell surface and concentrated on the microvilli and pseudopodia of the macrophages. The effects of LPS on expression of the integrins were dose and time related. VLA-5 expression was increased after 30 min of stimulation by LPS, suggesting that LPS may induce rapid secretion of the integrin. However, incubations with LPS longer than 30 min decreased VLA-5 expression in a dose-dependent pattern. LPS also caused dose-related decreases in the expression of VLA-3 and VLA-2 integrins and increases of intracellular FN 24 h after stimulation. The results suggest that a prolonged exposure to LPS may impede VLA integrin-mediated migration and result in local accumulation of macrophages in the lung.

Autoradiography↗

Partial protection against malaria by immunization with Leishmania enriettii expressing the Plasmodium yoelii circumsporozoite protein.

Since infection with Leishmania species induces CD4+ and CD8+ anti-leishmania T cells, we assessed protection against malaria by immunization with Leishmania enriettii transfected with the gene encoding the Plasmodium yoelii circumsporozoite protein (PyCSP). The recombinant plasmid appeared to be a circular episome in the host cells. Reverse transcription PCR showed that the PyCSP was trans-spliced by the addition of the 39-bp spliced leader of L. enriettii at its 5' end. The transfectant expressed a protein in a pattern similar to that found in the sporozoite itself. Immunofluorescence and immunoelectron microscopy indicated that PyCSP was abundantly expressed on the surface of the parasite. Mice immunized with the transfectant produced antibodies to sporozoites, had a delay in onset of parasitemia after challenge, and 4 of 22 (18%) were completely protected. The protected mice had cytotoxic T lymphocytes against the PyCSP. Immunization with recombinant vaccinia, Salmonella typhimurium, and pseudorabies virus expressing the PyCSP induces excellent immune responses, but has not been shown to protect against challenge. Thus, the modest protection found in these initial studies represents a step forward. After further work Leishmania may prove to be an important live vector vaccine system for induction of protective immune responses.

Amino Acid Sequence↗