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Biomedical subjects

M Kondo

Publications and source records attributed to M Kondo.

At least 433 records · Page 24Linked to original sources

Biodegradation of aniline, anthracene, chlornitrophen, fenitrothion and linear alkylbenzene sulphonate in pond water.

Biodegradation of five chemicals (aniline, anthracene, chlornitrophen (CNP), fenitrothion (FNT) and linear alkylbenzene sulphonate (LAS)) by aquatic bacteria in three different types of ponds was determined according to the cultivation method developed by this group. The degradability toward these chemicals was varied among the ponds, except for LAS which was decomposed well in all samples. Higher degradability towards the two agrochemicals, CNT and FNT, was found in the pond surrounded by paddy fields, whereas aniline and anthracene were decomposed more rapidly in the pond located in the industrial area. Water from the pond in the botanical garden, with the least exposure to any chemicals, exhibited the lowest degradation toward all chemicals tested. There was no significant seasonal variation in the biodegradation of chemicals in these ponds. It was deduced that biodegradability toward certain chemicals could be a result of acclimatization of the microbial community by chemical contamination present and past, suggesting the possible use of biodegradation profiles as an indicator for chemical pollution in the aquatic environment.

Biodegradation, Environmental↗

Characteristics of body composition and muscle strength in college Sumo wrestlers.

The purpose of this study was to investigate the characteristics of body composition and force generation capacity in college Sumo wrestlers (N=13, age=19.8+/-0.3 yr, stature= 178.5+/-1.6 cm, body mass = 111.2+/-3.8 kg, X+/-SE) in comparison with untrained males (N=18, 20.3+/-0.2 yr, 170.1+/-1.7 cm, 59.2+/-1.4 kg). The Sumo wrestlers had significantly higher average values in relative fat mass (24.8+/-1.0%) and fat-free mass (83.3+/-2.0 kg), estimated by an underwater weighing method, than the untrained subjects (relative fat mass = 12.9+/-0.1 %, fat-free mass = 51.5+/-1.3 kg). Moreover, the Sumo wrestlers had 1.7 to 1.9 times greater cross-sectional areas (CSAs) of elbow flexors and extensors and knee extensors, determined by a B-mode ultrasound technique, compared to those of the untrained subjects. Force values produced during elbow flexion and extension and knee extensions tasks under isokinetic contraction mode at constant velocities of 1.05, 3.14 and 5.24 rad x s(-1) were significantly higher in the Sumo wrestlers than in the untrained subjects. The force value in all test conditions was significantly correlated to the related-muscle CSA, r=0.611-0.910 (p<0.05). The difference between the two groups in force per unit CSA (F x CSA[-1]) during elbow flexion was not significant at all test velocities. However, the Sumo wrestlers showed significantly lower F x CSA(-1) values in elbow extension at 5.24 rad x s(-1) and in knee extension at all test velocities. Thus, the body composition of the Sumo wrestlers was characterized by a high fat content and a large fat-free mass. Moreover, the Sumo wrestlers had considerably larger muscle CSAs of limbs than the untrained subjects. For the Sumo wrestlers, however, force output of the muscles with a pennate structure were not proportional to their CSAs, particularly in knee extensors.

Adipose Tissue↗

Determination of interferon-alpha2 allele composition in the genomic DNA from healthy volunteers and leukemic patients in Japan.

The three interferon-alpha2 (IFN-alpha2) sequences identified to date differ from each other in just two nucleotide positions, both of which result in changes in amino acids. Thus, the mature IFN-alpha2a protein product is characterized by a lysine residue at position 23 (AAA) and a histidine at position 34 (CAA), IFN-alpha2b has an arginine at position 23 (AGA) and histidine at position 34 (CAT), and IFN-alpha2c has arginine residues at both positions 23 (AGA) and 34 (CGT). These nucleotide variations in the DNA sequence can be distinguished by selective restriction enzyme analysis. We studied the distributions of the three IFN-alpha2 variants by analyzing chromosomal DNA from 103 Japanese volunteers and 33 patients with hematologic disorders. Fragments of 238 bp and 617 bp of the IFN-alpha2 gene containing codons 23 and 34 were amplified by PCR using specific primers, and the PCR products were analyzed with specific restriction nucleases to identify the IFN-alpha2 variant sequences. Only IFN-alpha2b gene was detected in normal volunteers, and no IFN-alpha2a gene was detected in Japanese subjects. However, IFN-alpha2c was detected in 4 of 33 (12.1%) patients with leukemia.

Adult↗

Hydroxypyruvate reductase with a carboxy-terminal targeting signal to microbodies is expressed in Arabidopsis.

Five Arabidopsis EST cDNA clones of hydroxypyruvate reductase (HPR), a photorespiratory enzyme in leaf peroxisomes, were sequenced. Deduced amino acid sequences revealed that HPR in Arabidopsis contained the carboxy-terminal targeting signal to microbodies. Nucleotide sequence analysis showed that the cDNA with the longest insert contained an open reading frame of 1,158 bp which encoded a polypeptide with 386 amino acids with a calculated molecular mass of 42,251 Da. A Southern blot analysis suggested that the Arabidopsis HPR gene, like that of the pumpkin HPR gene, exists as a single copy. Two kinds of pumpkin HPR mRNA might be produced from a single gene by alternative splicing, but the structure of the genomic DNA indicated that the Arabidopsis HPR gene did not undergo alternative splicing. We detected a polypeptide with a molecular mass of 42 kDa in green leaves of Arabidopsis using an HPR-specific antibody. Immunoelectron microscopy revealed that Arabidopsis HPR protein was exclusively localized in leaf peroxisomes in green leaves. These results indicate that HPR is expressed in a form with a carboxy-terminal targeting signal to microbodies and is localized in microbodies in Arabidopsis, suggesting that the differences in the gene structure and the regulation of gene expression of HPR are probably due to species-specific differences in plants.

Alcohol Oxidoreductases↗

Changes in targeting efficiencies of proteins to plant microbodies caused by amino acid substitutions in the carboxy-terminal tripeptide.

It has been demonstrated that the carboxyl terminus of microbody enzymes functions as a targeting signal to microbodies in higher plants. We have examined an ability of 24 carboxy-terminal amino acid sequences to facilitate the transport of a cytosolic passenger protein, beta-glucuronidase, into microbodies in green cotyledonary cells of transgenic Arabidopsis. Immunoelectron microscopic analysis revealed that carboxy-terminal tripeptide sequences of the form [C/A/S/P]-[K/R]-[I/L/M] function as a microbody-targeting signal, although tripeptides with proline at the first amino acid position and isoleucine at the carboxyl terminus show weak targeting efficiencies. All known microbody enzymes that are synthesized in a form similar in size to the mature molecule, except catalase, contain one of these tripeptide sequences at their carboxyl terminus.

Amino Acids↗

beta-Glucosidase in the indigo plant: intracellular localization and tissue specific expression in leaves.

beta-Glucosidase of indigo plant (Polygonum tinctorium) has a high substrate specificity for indican (indoxyl beta-D-glucoside). To examine the localization of this beta-glucosidase, we fractionated the cells of the leaves and analysed them immunocytochemically. Immunoelectron micrographs with specific antibodies against the beta-glucosidase clearly showed that the beta-glucosidase was localized in the stroma of the chloroplasts in mesophyll cells, but not in the thylakoid membrane. Chloroplasts were isolated from the crude homogenate of the fresh leaves by Percoll density gradient centrifugation and then subjected to suborganellar fractionation. beta-Glucosidase activity was specifically detected in the stromal fraction, but not in the thylakoid membrane. This was also supported by the result of an immunoblot of the fraction with anti-beta-glucosidase antibodies. The beta-glucosidase was immunocytochemically localized in the chloroplasts of mesophyll cells, but not in any chloroplasts in marginal cells of the vascular bundle or epidermal cells; ribulose 1,5-bisphosphate carboxylase (Rubisco), a typical stromal protein, was observed in all chloroplasts in these cells. These results suggest that beta-glucosidase is tissue specific in its expression in the leaves of the indigo plant.

Plant Leaves↗

Role of superoxide and nitric oxide in platelet-activating factor-induced acute lung injury, hypotension, and mortality in rats.

OBJECTIVE: To investigate the role of superoxide and nitric oxide in platelet-activating factor-induced acute lung injury, hypotension, and mortality. DESIGN: Prospective, randomized, controlled, experimental study. SETTING: University research laboratory. SUBJECTS: Anesthetized male Wistar rats (180 to 220 g) were studied. INTERVENTIONS: In the first set of experiments, animals were divided into three groups. Group 1 received platelet-activating factor (2 microg/kg i.v.). Group 2 received recombinant human superoxide dismutase (50,000 U/kg i.v.) 30 mins before platelet-activating factor injection. Group 3 received vehicle agents. In the second set of experiments, animals were divided into six groups that received N(G)-nitro-L-arginine (L-NNA), a selective inhibitor of nitric oxide synthesis, or L-arginine, the physiologic precursor of nitric oxide synthesis: a) vehicles (i.v.); b) vehicle plus L-arginine (100 mg/kg i.v.); c) vehicle plus L-NNA (10 mg/kg i.v.); d) vehicle plus platelet-activating factor (2 microg/kg i.v.); e) L-arginine plus platelet-activating factor; and f) L-NNA plus platelet-activating factor. The first intravenous administration was given 5 mins before the second intravenous injection for each group. MEASUREMENTS AND MAIN RESULTS: In the first set of experiments, vascular labeling with Monastral blue B demonstrated diffuse microvascular injury in the alveolar capillary beds 2 hrs after platelet-activating factor challenge. Thiobarbituric acid-reactive substances in the lung significantly increased at 2 hrs after platelet-activating factor injection. Platelet-activating factor treatment also resulted in an increased concentration of total protein, albumin, and Evans blue dye in bronchoalveolar lavage fluid at 2 hrs after administration, suggesting platelet-activating factor induction of increased alveolar permeability. The platelet-activating factor-induced alveolar microvascular injury, lipid peroxidation, and increased alveolar permeability were inhibited by pretreatment with recombinant human superoxide dismutase. Although L-NNA alone did not affect alveolar permeability in the second set of experiments, L-NNA treatment before platelet-activating factor challenge significantly aggravated platelet-activating factor-induced increased alveolar permeability 2 hrs after platelet-activating factor challenge. Platelet-activating factor also produced a rapid decrease in blood pressure that was not ameliorated by treatment with L-NNA. However, L-NNA pretreatment was associated with a significant increase in platelet-activating factor-caused mortality within 6 hrs. All rats survived with L-arginine treatment before platelet-activating factor challenge. L-NNA treatment decreased nitrate/nitrite concentration, an index of total nitric oxide production, in plasma. CONCLUSIONS: These results indicate that superoxide, the derived active oxygen species, and lipid peroxidation are implicated in the pathogenesis of platelet-activating factor-induced acute lung injury. Nitric oxide does not play a major role in platelet-activating factor-induced hypotension. Nitric oxide appears to play a protective role in the acute lung injury and mortality induced by platelet-activating factor.

Animals↗

Monochloramine-induced cell growth inhibition and apoptosis in a rat gastric mucosal cell line.

Recent studies have indicated that monochloramine (NH2Cl), a reaction product of NH3 and hypochlorous acid, is involved in the pathogenesis of Helicobacter pylori-associated gastric mucosal damage, but how NH2Cl contributes to lesions is unclear. In the present study, the effects of NH2Cl on mucosal cell growth and the cell cycle were evaluated in vitro using a normal rat gastric mucosal cell line RGM-1. Cell viability was assessed by the Trypan Blue dye exclusion test and cell cycle patterns were determined by DNA labeling with propidium iodide and flow cytometric quantification. NH2Cl inhibited the growth of RGM-1 cells in a concentration-dependent manner. Exposure of cells to NH2Cl caused a time- and dose-dependent loss of G1-phase cells with accumulation of G2/M-phase cells, and produced a fraction of subdiploid cells with oligonucleosomal DNA degradation characteristic of apoptosis. NH2Cl-induced apoptosis was confirmed by fluorescent microscopy with Hoechst 33342 and propidium iodide. These results suggest that NH2Cl inhibits gastric mucosal cell growth and induces apoptosis in RGM-1 cells, events that may be important in gastric mucosal damage or atrophy induced by H. pylori infection.

Animals↗

Interactions of neutrophils and endothelial cells under low flow conditions in vitro.

The interactions of polymorphonuclear leukocytes (PMN) and endothelial cells are modulated by adhesion molecules, inflammatory cytokines, and shear stress. We investigated the changes in PMN-endothelial cell interactions induced by interleukin (IL)-1 beta under low flow conditions. PMN were isolated from the venous blood of healthy adults, and endothelial cells were obtained from human umbilical veins. The number of PMN that adhered to the endothelial cells monolayer that was treated with IL-1 increased significantly at shear stresses from .5 to 4.0 dyn/cm2 as compared with untreated endothelial cells. Anti-intercellular adhesion molecule (ICAM)-1 monoclonal antibody (mAb), anti-E-selectin mAb, and anti-CD18 mAb each significantly inhibited the increase in PMN adherence induced by IL-1 at a low shear stress (1.0 dyn/cm2). Anti-CD18 mAb significantly reduced the number of PMN that migrated through the endothelial monolayer by blocking the adherence of PMN to the luminal surface of the endothelial cells, as well as their transendothelial migration. In contrast, anti-ICAM-1 and anti-E-selectin mAb each reduced the number of PMN that migrated by reducing the number of PMN that adhered to the luminal surface without significantly influencing the percent of the adherent PMN that had migrated. Although anti-L-selectin mAb reduced the adherence and migration of PMN, these effects were not statistically significant. These results indicated that under low flow conditions, as well as in the nonflow state, PMN-endothelial cell interactions were elicited via CD11/CD18 and ICAM-1 without the involvement of selectins.

Adult↗

Expression of a vacuolar protein (VP24) in anthocyanin-producing cells of sweet potato in suspension culture.

VP24, an abundant protein of 24 kD, was found to accumulate in the anthocyanin-containing vacuoles of cells of sweet potato (Ipomoea batatas) in suspension culture. Light-induced expression of VP24 was analyzed by immunoblotting in three different cell lines that produced anthocyanins at different rates. The expression of VP24 was closely correlated with the accumulation of anthocyanin in these cell lines. Immunocytochemical detection of VP24 with specific antibodies on thin sections showed that VP24 was localized in the intravacuolar pigmented globules (cyanoplasts) in the anthocyanin-containing vacuoles and not in the tonoplast. No VP24 immunogold labeling was detected in the vacuoles of the cell line that does not produce anthocyanin. We suggest that VP24 may be involved in the formation of the cyanoplast via an interaction with anthocyanin, and that it may play an important role in the trapping in vacuoles of large amounts of anthocyanins that have been transported into these vacuoles.

Anthocyanins↗

The role of nitric oxide in hyperaemic response to flicker in the retina and optic nerve in cats.

PURPOSE: To elucidate the role of nitric oxide (NO) in the eye and the flicker-induced vascular response. METHODS: The blood flow in the retina and different parts of the optic nerve was compared in cats treated with the NO-synthase blocker, N(G)-nitro-L-arginine methyl ester and in control animals. In both groups, one of the eyes was dark-adapted, the other was subjected to 8 Hz flickering light. The regional blood flow was measured with the microsphere method. RESULTS: In control animals, flickering light increased blood flow in the retina and optic nerve head by 39% and 256%, respectively. Pretreatment with N(G)-nitro-L-arginine methyl ester prevented this increase in retinal blood flow and markedly reduced the blood flow in the optic nerve heads. CONCLUSION: NO release may mediate much of the vasodilating effect of flicker in cats, and play a role in maintaining normal vascular tone in the optic nerve head.

Animals↗

[Simple method for typing human immunodeficiency virus type I (B, E) by PCR and chronological change of distribution of subtype (B to E) in Japan].

We analyzed the clade distribution of B and E in HIV-1 isolates in Japan by a nested PCR method using 5'-CCCACAAGATTTAAATATG-3' of the gag gene as clade B primer and 5'-CCCACAAGATTTAAACTCC-3' of the gag gene as clade E primer. Seventy-two anti-HIV-1 confirmatory positive serum samples were collected during a period of 1991-1996 in two hospitals in Yokohama City. Peripheral blood mononuclear cells were obtained from the buffy coat of these samples and extracted DNA were used for nested PCR. The 72 cases comprised of 11 Japanese hemophiliacs, 14 Japanese male homosexuals, 19 Japanese male heterosexuals, 5 Japanese female heterosexuals and 23 Thai female heterosexuals. Of these 36 were clade B and 35 were clade E and one case showed positive PCR results for both B and E primers. Almost all male Japanese hemophiliacs and homosexuals in our sample have clade B, while the female Thai heterosexuals have clade E, irrespective of the year of isolation. As for Japanese male heterosexuals, through 1993, clade B was predominant but since 1994, the predominate clade switched to clade E. Although the number of Japanese female heterosexuals in our sample is small, clade B was isolated in 2 cases even after 1994.

Female↗

Lipopolysaccharide-induced goblet cell hypersecretion in the guinea pig trachea: inhibition by macrolides.

We studied the effects of macrolides on lipopolysaccharide (LPS)-induced airway goblet cell secretion in the guinea pig trachea. The goblet cell secretion was assessed in histological sections of the tracheal mucosa stained with alcian blue and periodic acid Schiff by arbitrarily determining mucus score, which is inversely related to the magnitude of mucus discharge. Inhalation of Escherichia coli LPS (5 mg/kg) caused a time-dependent decrease in mucus score, with the maximal response being from 542 +/- 49 to 92 +/- 20 arbitrary units (P < 0.001) after 3 h, which was accompanied by an increase in the number of neutrophils in the tracheal mucosa. The LPS-induced mucus discharge was inhibited by oral clarithromycin and erythromycin in a dose-dependent manner (5 and 10 mg/kg), whereas amoxicillin and cefaclor had no effect. Each dose of clarithromycin and erythromycin, but not amoxicillin or cefaclor, likewise attenuated the LPS-induced recruitment of neutrophils. These results suggest that LPS stimulates goblet cell secretion and neutrophil accumulation in the airways and that macrolides may be of value in protecting against neutrophil-associated airway hypersecretion.

Animals↗

Effect of Z-103 on TNB-induced colitis in rats.

Z-103 is a chelate compound consisting of zinc ion and L-carnosine. In this study, we investigated the protective effect of Z-103 against colonic damage induced by 2,4,6-trinitrobenzene sulfonic acid (TNB) in rats. Colonic inflammation was induced by administering TNB dissolved in 50% ethanol (120 mg/ml) in male Wistar rats (total volume of 0.25 ml per rat) following a 48-hour fast. After the administration of TNB, Z-103 was given at a dose of 30 mg/kg per rat for 1 week. A second group of rats received sulfasalazine (SASP) at 300 mg/kg and a third group of rats received 30 mg/kg of ZnSO4 for 1 week. Colonic inflammation was assessed 1 week following TNB administration. Both macro- and microscopic evaluation showed that the inflammatory responses induced by TNB were reduced by treatment with Z-103, SASP and ZnSO4. The score (graded from 0 to 5 according to the macroscopic lesions) and colonic wet weight (distal 8 cm of the colon) were significantly decreased by treatment with Z-103, SASP and ZnSO4. The increase in thiobarbituric acid-reactive substances in the colonic mucosa following TNB administration were inhibited in the Z-103 and SASP groups. These results suggest that Z-103 is as effective against TNB-induced colitis as SASP.

Animals↗

Characterization of a liver metastatic variant of murine colon 26 carcinoma cells.

Intraportal vein injection of highly metastatic L5 cells consistently resulted in liver metastases (increases in the number of tumor colonies in the liver), whereas inoculation of P cells rarely did. L5 cells invaded the basement membrane Matrigel in greater numbers than did P cells, suggesting that the metastatic potential of L5 cells is partly related to enhanced invasive properties. The enhanced adhesion of L5 cells to fibronectin-, laminin- and Matrigel-coated substrates, as well as their haptotactic migration to fribronectin, may be associated with the preferential expression of VLA-2 and VLA-4 integrins on the surface of these cells detected by flow cytometry. Gelatin zymograms showed that the degradative activity of 72-kD gelatinases was greater in L5 cells than P cells. These results indicate that, in addition to adhesiveness and motility, the invasive ability of L5 cells may also be attributed to enhanced gelatinolytic activity. L5 cells grew more rapidly than P cells in vitro. Thus, an experimental model using highly metastatic colon 26 L5 cells would be useful for analyzing the molecular mechanism of liver metastasis and for evaluating the efficacy of treatment of occult micrometastases which may already have been disseminated at the time of surgery.

Adenocarcinoma↗

Bactericidal activities of povidone-iodine against Mycobacterium.

Three standard strains of Mycobacterium (M. tuberculosis H37Rv, M. avium ATCC15769 and M. kansasii ATCC12478) and 15 clinical isolates of Mycobacterium (7 M. tuberculosis, 2 M. avium, 3 M. kansasii, 1 M. intracellulare, 1 M. chelonae subsp. abscessus and 1 M. gordonae) were selected in order to study the bactericidal activities of povidone-iodine (PVP-I) drug substance and a commercially available PVP-I solution (Isodine solution) against Mycobacterium. After the bacilli had been exposed to the disinfectant solution at concentrations of 0.1 or 0.02% with 2% human serum for various incubation periods from 30 to 120 s, the PVP-I drug substance was inactivated by addition of 0.5% sodium thiosulfate. In the case of the commercially available PVP-I solution, a mixture of 10% Tween 80, 3% soybean lecithin and 0.5% sodium thiosulfate was used as inactivator. It was demonstrated that the 3 standard strains were completely inactivated within 30 s by 0.1% PVP-I drug substance and that the 15 clinical isolates were almost killed by 0.1% commercially available PVP-I solution within 60 s. As a result, the commercially available PVP-I product appeared to be a useful agent as disinfectant against all the tested species of Mycobacterium.

Anti-Infective Agents, Local↗