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Biomedical subjects

H J Koh

Publications and source records attributed to H J Koh.

At least 19 recordsLinked to original sources

Bran extracts from pigmented rice seeds inhibit tumor promotion in lymphoblastoid B cells by phorbol ester.

Using flow cytometry, we quantitatively evaluated anti-tumor-promoting activity of rice bran extracts by measuring inhibition of Epstein-Barr virus early-antigen activation (EBV-EA) induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). This assay measures anti-tumor-promoting activity and cytotoxicity of target substances using the same batch of cells. Using this short-term procedure, we have determined the anti-tumor-promoting activity of 70% ethanol-water extracts of bran (outer layer) from seeds of five pigmented rice cultivars: Jumlalocal-1, DZ 78, Elwee, LK1-3-6-12-1-1, and LK1A-2-12-1-1. The results show that, compared to an extract from the non-pigmented white cooking rice variety Chuchung, the extracts from the pigmented varieties strongly inhibited phorbol ester-induced tumor promotion in marmoset lymphoblastoid cells B95-8 in vitro. These findings may facilitate development and use of new health-promoting rice varieties.

Animals↗

Role of NADP+-dependent isocitrate dehydrogenase (NADP+-ICDH) on cellular defence against oxidative injury by gamma-rays.

PURPOSE: To investigate the regulation of NADPH-producing isocitrate dehydrogenase (ICDH) in cytosol (IDPc) and mitochondria (IDPm) upon gamma-ray irradiation, and the roles of IDPc and IDPm in the protection against cellular damage induced by gamma-ray irradiation. MATERIALS AND METHODS: Changes of IDPc and IDPm proteins upon gamma-ray irradiation to NIH3T3 cells were analysed by immunoblotting. To increase or decrease the expression of IDPc or IDPm, NIH3T3 cells were stably transfected with mouse IDPc or IDPm cDNA in either the sense or the antisense direction. The transfected cells with either increased or decreased IDPc or IDPm were exposed to gamma-rays, and the levels of reactive oxygen species generation, protein oxidation and lipid peroxidation were measured. RESULTS: Both IDPc and IDPm activities were induced by gamma-ray in NIH3T3 cells. Cells with decreased expression of IDPc or IDPm had elevated reactive oxygen species generation, lipid peroxidation and protein oxidation. Conversely, overproduction of IDPc or IDPm protein partially protected the cells from oxidative damage induced by gamma-ray irradiation. CONCLUSIONS: The protective role of IDPc and IDPm against gamma-ray-induced cellular damage can be attributed to elevated NADPH, reducing equivalents needed for recycling reduced glutathione in the cytosol and mitochondria. Thus, a primary biological function of the ICDHs may be production of NADPH, which is a prerequisite for some cellular defence systems against oxidative damage.

Animals↗

Effects of risperidone on information processing and attention in first-episode schizophrenia.

Risperidone appears to have a beneficial effect in several areas of cognitive function in schizophrenic patients. In previous studies, however, the clinical characteristics of the subjects differed between studies, and were heterogeneous even in single study. Most of the subjects were previously exposed to different kinds of neuroleptics and some of them were treatment-resistant. The aim of this study is to evaluate the effect of risperidone on attention and information processing in a homogeneous subgroup of schizophrenic patients, i.e. drug-naïve first-episode inpatients. In the patient group (n=17), cognitive tests and clinical assessments were performed before and after 8 weeks of risperidone treatment. The same cognitive tests were administered to the control group (n=24). The delay between test and retest was 8 weeks. Before treatment, the patient group performed significantly worse than control group on the tests measuring continuous attention, vigilance, and the speed of information processing. After treatment (the average dose of risperidone was 7.3mg/day), in spite of significant improvement of the clinical symptoms, the patients did not show any significant improvement or worsening in most of the items of these tests. The control group did not show any practice effect. These results suggest that first-episode schizophrenic patients have deficits in sustained attention and vigilance to visual stimuli, as well as the speed of information processing to visual and auditory stimuli, and these deficits are unrelated to clinical symptoms and remain stable during the early phase of treatment. This study did not receive pharmaceutical company financial support.

Adolescent↗

Enhanced expression of transglutaminase 2 in anterior polar cataracts and its induction by TGF-beta in vitro.

BACKGROUND/AIMS: Transglutaminase activity has long been implicated in the cataract formation. However, the precise mechanism of how it is produced and involved in this process remains unclear. Here the authors sought to examine whether transglutaminase 2 (TGase 2) is expressed in lens epithelial cells from patients with anterior polar cataracts, to determine whether TGase 2 expression is induced by transforming growth factor (TGF-beta) in cultured lens epithelial cells, and to determine whether TGase 2 participates in the crosslinking of fibronectin in lens epithelial cells in vitro. METHODS: Lens epithelial cells from anterior polar cataracts, nuclear cataracts, and non-cataractous clear lenses were examined for the expression of TGase 2 using reverse transcription-polymerase chain reaction, western blot analysis, and immunohistochemical analysis. The modulation of extracellular TGase 2 activity by TGF-beta was measured by the formation of fibronectin polymers and the incorporation of fluorescein cadaverine into extracellular matrix proteins. The effect of TGase 2 overexpression was analysed by immunofluorescence staining and western blot analysis of human lens epithelial (HLE) B-3 cells transiently transfected with TGase 2 gene. RESULTS: The expression of TGase 2 mRNA and its protein was markedly enhanced in lens epithelial cells from patients with anterior polar cataracts. Treatment of HLE B-3 cells with TGF-beta caused an increase in TGase 2 protein, its extracellular activity, and the crosslinking of fibronectin. Transient transfection of HLE B-3 cells with the TGase 2 gene led to the increased production of fibronectin monomers and polymers. CONCLUSIONS: This study shows that TGase 2 is overexpressed in lens epithelial cells from anterior polar cataracts and that TGF-beta may be a causative factor in the induction of TGase 2. The enhanced expression of TGase 2 might cause the accumulation and crosslinking of the extracellular matrix proteins and might play a part in anterior polar cataract development.

Aged↗

Structure-reactivity relationships in the pyridinolysis of N-methyl-N-arylcarbamoyl chlorides in dimethyl sulfoxide.

Nucleophilic substitution reactions of N-methyl-N-arylcarbamoyl chlorides (YC(6)H(4)N(CH(3))COCl) with pyridines (XC(5)H(4)N) have been investigated in dimethyl sulfoxide at 45.0 degrees C. A striking trend in the selectivity parameters is that they are constant within experimental errors, rho(X) = -2.25 +/- 0.03, beta(X) = 0.42 +/- 0.01, and rho(Y) = 1.10 +/- 0.06, with changing reactivities of the electrophiles (deltasigma(Y)) and nucleophiles (deltasigma(X)), respectively, and this leads to a vanishingly small cross-interaction constant, rho(XY) approximately equals beta(XY) approximately equals 0. The rate data can be expressed in the Ritchie N(+) type equation. Based on this and other results, the mechanism of nucleophile (pyridine) addition to the resonance- stabilized carbocation is proposed. It has been shown from the definition of beta(XY) (and rho(XY)) together with the Marcus equation that the high intrinsic barrier, DeltaG(0), in the intrinsic-barrier controlled reaction series is a prerequisite for such reactions in which the cross-interaction vanishes and the N(+) relationship holds.

Journal Article↗

Control of mitochondrial redox balance and cellular defense against oxidative damage by mitochondrial NADP+-dependent isocitrate dehydrogenase.

Mitochondria are the major organelles that produce reactive oxygen species (ROS) and the main target of ROS-induced damage as observed in various pathological states including aging. Production of NADPH required for the regeneration of glutathione in the mitochondria is critical for scavenging mitochondrial ROS through glutathione reductase and peroxidase systems. We investigated the role of mitochondrial NADP(+)-dependent isocitrate dehydrogenase (IDPm) in controlling the mitochondrial redox balance and subsequent cellular defense against oxidative damage. We demonstrate in this report that IDPm is induced by ROS and that decreased expression of IDPm markedly elevates the ROS generation, DNA fragmentation, lipid peroxidation, and concurrent mitochondrial damage with a significant reduction in ATP level. Conversely, overproduction of IDPm protein efficiently protected the cells from ROS-induced damage. The protective role of IDPm against oxidative damage may be attributed to increased levels of a reducing equivalent, NADPH, needed for regeneration of glutathione in the mitochondria. Our results strongly indicate that IDPm is a major NADPH producer in the mitochondria and thus plays a key role in cellular defense against oxidative stress-induced damage.

3T3 Cells↗

Translimbal approach for intravitreal injection in endophthalmitis after phacoemulsification.

We describe a corneal limbal technique of intravitreal injection for use in cases in which it is difficult to confirm the position of the needle. Using this translimbal approach, the needle's position is easily identified and the intravitreal injection can be given using topical anesthesia. This technique is an option in eyes with a cloudy cornea or a large iridectomy.

Adult↗

TGF-betas synthesized by RPE cells have autocrine activity on mesenchymal transformation and cell proliferation.

The present study investigated the effects of transforming growth factor (TGF)-beta on retinal pigment epithelial (RPE) transformation in a simplified model and also whether or not TGF-beta exhibits similar proliferation effects on transformed RPE cells that it has on primary RPE cells. Furthermore, we examined the cell proliferation effects of RPE-conditioned medium (CM). A vertical wound measuring 2 mm in diameter was made on primary RPE monolayers. The expression of alpha-smooth muscle actin (SMA) by the cells located at the wound edges was observed using a confocal microscope under immunofluorescent staining. Cell proliferation was measured by incorporating 3H-thymidine into DNA. The presence of alpha-SMA was observed in the cells within the wound after treatment with TGF-beta2, while negative expression was observed in control cells. TGF-betas inhibited the proliferation of the primary cultures of RPE cells in a dose-dependent manner, but the spindle-shaped late-passaged RPE cells were not inhibited by these growth factors. The medium conditioned by RPE cells stimulated the proliferation of subconjunctival fibroblasts and inhibited the proliferation of primary RPE cells, in a manner similar to TGF-beta. These findings demonstrate that TGF-beta-stimulated RPE cells may evoke proliferative vitreoretinopathy through mesenchymal transformation and cell proliferation.

Actins↗

Scleral fixation technique using 2 corneal tunnels for a dislocated intraocular lens.

Several techniques are used to reposition dislocated intraocular lenses (IOLs). Most place a suture loop around the end of the haptic. However, in cases of a dislocated capsular bag containing the IOL, a dislocated IOL with a large haptic, or a miotic pupil, it is not easy to see the haptic ends to place the suture loop. We describe a scleral fixation technique that uses 2 corneal tunnels. A double-armed 10-0 polypropylene suture loop can be introduced through 1 corneal tunnel and placed around any accessible part of the haptic with the help of a bent 26 gauge needle. This modified technique is an easy and effective way to reposition the IOL.

Biocompatible Materials↗

Mechanism of regulation of Na+ transport by angiotensin II in primary renal cells.

BACKGROUND: Angiotensin II (Ang II) has a dose-dependent, biphasic effect on the activity of the Na+/H+ antiport system in the renal proximal tubule (RPT). The aim of the present study was to further delineate the signaling pathways involved in Ang II action. METHODS: To examine Ang II signaling, 22Na+ uptake studies were conducted with a primary rabbit RPT cell culture system. The activation of phospholipase A2 (PLA2) was assessed by measuring the release of [3H]-arachidonic acid (AA), and changes in intracellular calcium levels were determined by means of confocal microscopy. RESULTS: Low dosages of Ang II (<10-10 mol/L) stimulated Na+ uptake, whereas high dosages of Ang II (>10-10 mol/L) inhibited Na+ uptake. Ang II (>10-10 mol/L) also caused an increase in AA release associated with an increase in intracellular calcium. Not only did exogenous AA inhibit Na+ uptake, but two PLA2 inhibitors (mepacrine and AACOCF3) blocked the Ang II-mediated inhibition of Na+ uptake. However, the cytochrome P450-dependent epoxygenase inhibitor econazole also blocked the Ang II-induced inhibition of Na+ uptake. Inhibition of Na+ uptake was obtained by the metabolic product of the epoxygenase 5,6-EET. In turn, the inhibitory effect of 5,6-EET was blocked by indomethacin. CONCLUSIONS: The results indicate the involvement of a calcium-dependent PLA2 in mediating the inhibitory effect of Ang II on Na+ uptake. The AA, which is released following PLA2 activation, acts indirectly, through its own metabolism, via a cytochrome P450 epoxygenase pathway and ultimately cyclooxygenase itself.

Angiotensin II↗

Effect of IOP elevation on matrix metalloproteinase-2 in rabbit anterior chamber.

To investigate changes in the level of matrix metalloproteinase-2 (MMP-2) in the anterior chamber of rabbit with intraocular pressure (IOP) elevation. The IOP was elevated with scleral encircling in 12 rabbits. In the control group (4 rabbits), IOP was not changed after scleral encircling, and in group 1 (4 rabbits) and 2 (4 rabbits), IOP was elevated about 10 and 20 mmHg respectively after scleral encircling. At 2 days after scleral encircling, aqueous sampling was performed and levels of MMP-2 were checked by Western blots and gelatin zymograms. The greater the IOP elevation, the more MMP-2 expression in the anterior chamber by Western blots and gelatin zymograms. The increase in MMP-2 expression in response to IOP elevation may have important implications for the IOP feedback control mechanism.

Animals↗

Inadvertent topical exposure to isocyanates caused damage to the entire eyeball.

Isocyanates are part of a group of important chemicals necessary in the production of adhesives, synthetic rubbers, and a variety of plastics. They are known to have minimal toxic effects when administered locally. However, we experienced a case of damage involving the entire eyeball in a person who accidentally exposed his eye to isocyanates. The patient presented with inflamed conjunctiva, and shrunken cornea and sclera, with focal atrophic changes in the iris. The lens and vitreous were opacified. After removal of the lens and vitreous, there were large areas of atrophic retina and areas of retinal necrosis with holes. We found that locally absorbed isocyanates can cause damage to the entire eyeball. Therefore, we recommend that if there is any evidence of isocyanate penetration, early vitrectomy should be performed to help in determining the extent of retina damage and to decrease the amount of toxic substance in the vitreous.

Accidents, Occupational↗

Massive suprachoroidal hemorrhage with retinal and vitreous incarceration; a vitreoretinal surgical approach.

Suprachoroidal hemorrhage(SH) may cause the expulsion of the intraocular contents. Vitreous incarceration in the wound and retinal detachment with SH are extremely poor prognostic signs. Treatment modalities depend on the severity of eye damage. This particular patient had "kissing" hemorrhagic choroidal detachment which completely filled the vitreous cavity after cataract surgery. It seemed to be inoperable. Secondary surgery was delayed 3 days to lower IOP to normal levels. The eye underwent anterior drainage sclerotomy under constantly-maintained limbal or pars plana infusion fluid line pressure. The authors performed a pars plana vitrectomy, followed by perfluorocarbon liquid injection and a silicone oil tamponade. After this surgical approach, the patient attained an attached retina and a visual acuity of 5/200 at the 3 month follow-up.

Aged↗

Identification and functional characterization of a novel, tissue-specific NAD(+)-dependent isocitrate dehydrogenase beta subunit isoform.

To understand the interactions and functional role of each of the three mitochondrial NAD(+)-dependent isocitrate dehydrogenase (IDH) subunits (alpha, beta, and gamma), we have characterized human cDNAs encoding two beta isoforms (beta(1) and beta(2)) and the gamma subunit. Analysis of deduced amino acid sequences revealed that beta(1) and beta(2) encode 349 and 354 amino acids, respectively, and the two isoforms only differ in the most carboxyl 28 amino acids. The gamma cDNA encodes 354 amino acids and is almost identical to monkey IDHgamma. Northern analyses revealed that the smaller beta(2) transcript (1.3 kilobases) is primarily expressed in heart and skeletal muscle, whereas the larger beta(1) mRNA (1.6 kilobases) is prevalent in nonmuscle tissues. Sequence analysis of the IDHbeta gene indicates that the difference in the C-terminal 28 amino acids between beta(1) and beta(2) proteins results from alternative splicing of a single transcript. Among the various combinations of human IDH subunits co-expressed in bacteria, alphabetagamma, alphabeta, and alphagamma combinations exhibited significant amounts of IDH activity, whereas subunits produced alone and betagamma showed no detectable activity. These data suggest that the alpha is the catalytic subunit and that at least one of the other two subunits plays an essential supporting role for activity. Substitution of beta(1) with beta(2) in the co-expression system lowered the pH optimum for IDH activity from 8.0 to 7.6. This difference in optimal pH was analogous to what was observed in mouse kidney and brain (beta(1) prevalent; optimal pH 8.0) versus heart (beta(2) prevalent; pH 7.6) mitochondria. Experiments with a specially designed splicing reporter construct stably transfected into HT1080 cells indicate that acidic conditions favor a splicing pattern responsible for the muscle- and heart-specific beta(2) isoform. Taken together, these data indicate a regulatory role of IDHbeta isoforms in determining the pH optimum for IDH activity through the tissue-specific alternative splicing.

Alternative Splicing↗

Radiation sensitivity of an Escherichia coli mutant lacking NADP+-dependent isocitrate dehydrogenase.

Ionizing radiation induces the production of reactive oxygen species, which play an important causative role in radiation damage. NADP+-dependent isocitrate dehydrogenase (ICDH) in Escherichia coli produces NADPH, an essential reducing equivalent for the antioxidant system. The protective role of ICDH against ionizing radiation in E. coli was investigated in wild-type and ICDH-deficient strains. Upon exposure to ionizing radiation, the viability was lower and the lipid peroxidation was higher in mutant cells compared to wild-type cells. Activities of key antioxidant enzymes such as superoxide dismutase, catalase, glutathione reductase, and glucose-6-phosphate dehydrogenase were decreased by irradiation in both cells. Results suggest that ICDH plays an important role as an antioxidant enzyme in cellular defense against ionizing radiation.

Antioxidants↗