Expression of intercellular adhesion molecule-1 on mouse mesangial cells and cytokines from mononuclear cells of IgA nephropathy.
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Animals↗
Biomedical subjects
M J Kim
Publications and source records attributed to M J Kim.
Bone marrow findings before and after treatment with recombinant human erythropoietin in chronic hemodialyzed patients.
The stimulating effect of rHuEPO on erythropoiesis has been shown in several studies, using bone marrow cell culture or animal models. To investigate the effect of rHuEPO on bone marrow findings in vivo, we studied the bone marrow cellularity, the myeloid: erythroid (M:E) ratio, an estimate of the number of megakaryocytes, any cytomorphologic or maturation abnormalities, and an estimate of the storage iron before and after 3 months of treatment with rHuEPO in 10 chronic hemodialyzed patients. Nine patients showed a slight or moderate decrease of erythropoiesis in bone marrow in comparison to normal bone marrow before rHuEPO treatment. The bone marrow cellularity was a mean of 28.5% and decreased in 8 out of 10 patients compared to normal values. However, megakaryopoiesis and granulopoiesis were normal. Three months of treatment with rHuEPO had increased erythropoiesis in all 10 patients, including one patient whose bone marrow proved to be normal erythropoiesis on baseline examination. The M:E ratio was significantly decreased from 4.0 +/- 1.2:1 to 2.4 +/- 1.1:1 (p < 0.005). The bone marrow cellularity was also increased in 9 patients, except in 1 patient whose specimen was inadequate for diagnosis, after 3 months of treatment with rHuEPO. On baseline examination of bone marrow, iron staining was undetectable in one, low in one, normal in 2 and high in 5 sections. According to grading, iron staining had decreased from 3.1 +/- 1.7 to 2.1 +/- 0.9 (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗
Evidences for possible involvement of Rhp51 protein in mitotic events including chromosome segregation.
To understand the role of the Rhp51 protein in Schizosaccharomyces pombe, we examined the phenotypes of the null mutant for the rhp51+ gene. Unlike Saccharomyces cerevisiae rad51 mutants, S. pombe rhp51 mutants (rhp51delta cells) displayed slow growth and heterogeneity in cell size, indicating perturbation of the cell cycle. Furthermore, many aberrant nuclear structures found in 4',6'-diamidino-2-phenylindole (DAPI)-stained rhp51delta cells and the caffeine hypersensitivity of the mutant cells suggested an involvement of the Rhp51 protein in normal chromosome segregation. These data suggested that the Rhp51 proteins were required for normal cell growth as well as a DNA repair pathway. Moreover, rhp51delta mutants showed a considerable sensitivity to ultraviolet (UV) light-irradiation as well as methyl methanesulfonate (MMS) treatment, indicating that the Rhp51 proteins are involved in both the active excision mechanism of UV-induced DNA damage and recombinational repair in S. pombe. Taken together, we suggest that the role(s) of the Rhp51 protein in S. pombe may be different from those of Rad51 in S. cerevisiae.
DNA Damage↗
A simple and efficient method for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe.
A simple and efficient procedure was described for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe. The present study demonstrated that the quality and the quantity of S. pombe RNA were increased by substituting phenol/chloroform mixture for phenol as a deproteinizing agent in the first vortexing step and using an ice bath instead of a dry ice-ethanol bath in the freezing step. Additionally, this protocol had the advantage of extracting total RNA without any degradation of S. pombe cells. Furthermore, the high amounts and quality of RNA extracted by this modified procedure enabled us to perform some experiments such as Northern blot, S1 mapping, primer extension, and reverse transcriptase reaction-polymerase chain reaction (RT-PCR) without further RNA purification. We suggest that this procedure is very useful to analyse primary structures and steady-state levels of RNA from S. pombe.
Base Sequence↗
Phosphatase-negative mutants of Legionella pneumophila and their behavior in mammalian cell infection.
Microbial phosphatases are known or suspected to play a role in the pathogenesis of several intracellular pathogens, including Legionella micdadei. Legionella pneumophila also possess phosphatase activities, but their possible roles in cellular infection are unknown. We generated mutants of a serogroup 1 isolate of L. pneumophila that lack the major phosphatase. Isolation of a Pho- mutant after random mutagenesis with transposon MudII4041 allowed us to dissociate the major alkaline phosphatase (pH optimum approximately 8) from a minor acid phosphatase activity. Both activities were concentrated in the bacterial periplasm. The gene encoding the major alkaline phosphatase (pho) was cloned by expression in E. coli and used to generate a site directed mutation in two L. pneumophila strains. Each parent-mutant pair was compared in a U937 cell tissue culture assay for capacity to infect, lyse, and grow within mammalian cells. Although the parental stains differed in their U937 cell cytopathicity, neither was significantly more infective than its Pho- derivative, suggesting that the alkaline phosphatase activity is not essential for cellular infection. Because they are not attenuated, Pho- mutants can be used to generate gene fusions with E. coli alkaline phosphatase to study and secretion and cellular infectivity in L. pneumophila.
Alkaline Phosphatase↗
Transendothelial transport of modified low-density lipoproteins.
Human umbilical vein endothelial cell monolayers were grown as monolayers on porous filters and their transcellular transport and degradation of 125I-labelled native and modified forms of LDL, supplied to either the intimal or the luminal face, were measured. Intact native, acetylated and oxidized LDL were all transported in both directions across the cell monolayers by receptor-independent mechanisms, and all forms of LDL were transported at similar rates. However, the mass of intact LDL transported from the intimal to the luminal face of the monolayer was always fourfold more than that transported in the opposite direction under similar conditions. In addition to LDL transport, endothelial cell monolayers also degraded native and modified forms of LDL by predominantly receptor-dependent routes, in that these could be inhibited (> 70%) by the addition of a 20-fold excess of the same form of (but unlabelled) LDL. The measured amounts of lipoprotein degraded were the same whether supplied to the intimal or the luminal face. Incubation of endothelial cells with oxidized LDL led to intracellular accumulation of a pool of macromolecular apo B which was apparently resistant to lysosomal proteolysis.
Biological Transport↗
Occupational asthma caused by two herb materials, Dioscorea batatas and Pinellia ternata.
There have been very few cases of occupational allergies caused by herb materials. In this paper, we report a case of occupational asthma and rhinitis caused by two herb materials, Dioscorea batatas and Pinellia ternata, which are known as Sanyak and Banha respectively. The patient had strong positive responses to grass pollens and cat fur, as well as to Sanyak and Banha extracts on skin-prick tests. Bronchoprovocation tests showed a dual asthmatic response to Sanyak whereas only an early asthmatic response to Banha. Serum specific IgE and IgG4 antibodies to Sanyak were detected by enzyme-linked immunosorbent assay (ELISA) but there was no specific IgE antibody binding to Banha. In order to further characterize the allergenic component of these extracts, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotting studies were also performed. Three IgE binding components (29,000, 57,000, 63,000 Da) were detected within the Sanyak extracts, but no IgE-binding component was noted within the Banha extracts. It is suggested that Sanyak can induce IgE-mediated bronchoconstriction in an exposed worker. Further studies are needed to investigate the pathogenetic mechanism triggered by Banha.
Adult↗
Peer review: when enough is enough.
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Faculty, Nursing↗
Antigenic relationship between mugwort and ragweed pollens by crossed immunoelectrophoresis.
Mugwort and ragweed pollens have been considered as important respiratory allergens in Korea. These two pollens are abundant in the air of Seoul from August through October. Many ragweed-sensitive patients have shown concurrent sensitivities to mugwort pollen. However the antigenic relationship between these two pollens has not been clarified. To observe the cross-reactivity between them, we developed polyclonal anti-mugwort and anti-ragweed antibodies by immunization on New Zealand white rabbits, and performed crossed immunoelectrophoresis(CIE) with two pollen extracts. Five precipitation lines were formed by mugwort and anti-mugwort antibody. One precipitation line was formed by ragweed and anti-ragweed antibody. There was no reaction from mugwort and anti-ragweed antibody, and from ragweed and anti-mugwort antibody. These results indicate that there is no cross-antigenicity between mugwort and ragweed pollens.
Animals↗
Ipsilateral synchronous renal cell carcinoma and transitional cell carcinoma.
The simultaneous occurrence of renal cell carcinoma(RCC) and transitional cell carcinoma(TCC) in the same kidney is unusual. We report a 53-year-old man with ipsilateral synchronous renal adenocarcinoma and renal pelvic transitional cell carcinoma with severe hypercalcemia and a huge staghorn calculus in the opposite kidney. The patient was admitted to the hospital because of left flank pain and intermittent fever which he had had for 2 months. Computerized tomography revealed a huge stone in the right kidney and a mass in the upper pole with an irregular calcified pelvis in the left enlarged kidney. Left radical nephrectomy was done. A section of the specimen revealed a renal cell carcinoma located at the upper pole and a papillary transitional cell carcinoma arising from the renal pelvis. This is a rare case of combined renal malignancies.
Carcinoma, Renal Cell↗
Tumor necrosis factor alpha from peripheral blood mononuclear cells of IgA nephropathy and mesangial cell proliferation.
BACKGROUND: To investigate the role of mononuclear cells and their products on rat mesangial cell proliferation, we evaluated the effect of peripheral blood mononuclear cells (PBMC) culture supernatant of patients with IgA nephropathy on rat mesangial cells 3[H]-thymidine uptake, and measured the concentration of cytokines in the supernatant to find out which cytokine in PBMC culture supernatant has a major influence on mesangial cell 3[H]-thymidine uptake. METHODS: In ten patients with primary IgA nephropathy and 10 normal controls, PBMC were cultured with PHA (20 micrograms/ml) and con-A (10 micrograms/ml) for 24 hours, and supernatants were collected and stored at -70 degrees C until tested. Mesangial cell was cultured for 48 hours at a concentration of 10(4) cells/well with 1:2 or 1:10 diluted PBMC culture supernatant. 3[H]-thymidine uptake into rat mesangial cell was assayed after 18 hour culture. The cytokine concentrations of PBMC culture supernatant were measured by radioimmunoassay. RESULTS: When 1:2 diluted PBMC culture supernatant was added, the supernatant of IgA nephropathy induced higher 3[H]-thymidine uptake into mesangial cell compared with that of normal controls. 3[H]-thymidine uptake induced by 1:2 diluted PBMC culture supernatant was significantly higher than that induced by 1:10 diluted supernatant of IgA nephropathy. The concentration of TNF-alpha in PBMC culture supernatant of the patients with IgA nephropathy was higher than that of normal controls and showed a good correlation with the mesangial cell 3[H]-thymidine uptake. CONCLUSION: It is suggested that the mononuclear cell of IgA nephropathy may have the intrinsic property to produce more TNF-alpha which may be mitogenic to mesangial cells when stimulated with mitogens, and it may be related to the mesangial cell proliferation in IgA nephropathy.
Adolescent↗
Effect of angiotensin converting enzyme inhibitor on collagen production by cultured mesangial cells.
Cultured mesangial cells (MC) express renin mRNA and generate angiotensin I, supporting the action of local renin-angiotensin system. Also angiotensin II may act like a growth factor and was reported to increase collagen production (CP) in cultured MC. Angiotensin converting enzyme inhibitor is suggested to attenuate development and advancement of glomerulosclerosis, mainly with its hemodynamic effects. Therefore, we investigated the direct effects of enalapril (E) on CP by cultured MC. Rat MC were cultured in DMEM media alone, or containing high glucose (HG: 25 mM) or soluble immune complex (IC) prepared with bovine gamma globulin (BGG) and anti-BGG, with or without E (0.2 microgram/ml). CP was determined after 24 h by [3H] proline incorporation method. E significantly reduced CP by 43% in medium as compared with control (C) (C: 37,210 +/- 4,200 vs C + E: 21,350 +/- 5,080 cpm/well, p < 0.01). CP in medium increased in the presence of HG (123% of C) or IC (147% of C), which was, however, prevented with E (HG + E: 105% of C, IC + E: 116% of C). There were no differences of CP in cell layer between C (3,490 +/- 220 cpm/well) and C + E (3,340 +/- 190 cpm/well), and also no changes after addition of E in HG or IC groups. In conclusion, E directly attenuates CP by MC, even in the presence of HG or IC, independently of its hemodynamic effects.
Angiotensin-Converting Enzyme Inhibitors↗
The expression of beta 1 integrin and intercellular adhesion molecule-1 on mesangial cells and its modulation in glomerulonephritis.
OBJECTIVES: This study was performed to observe the effects of conditioned media (CM) of cultured mononuclear cells (MNC) on the expression of beta 1 integrin and intercellular adhesion molecule(ICAM)-1 on mesangial cells and the modulation by TGF-beta, TNF alpha, or hydroxyl radical Also these were examined in anti-Thy mesangial proliferative GN (MsPGN) or puromycin aminonucleoside nephrosis(PAN). METHODS: The expression of beta 1 integrin and ICAM-1 on mesangial cells was examined by ELISA and the modulation by TGF-beta, TNF alpha, or hydroxyl radical was investigated by using neutralizing antibody and dimethylthiourea. [.OH] scavenger. RESULTS: The CM of MNC from Control suppressed the beta 1 integrin expression and thymidine incorporation and increased ICAM-1 expression on mesangial cells, compared to media alone. Especially the significant increase in ICAM-1 expression was reversed by anti-TNF alpha antibody. beta 1 integrin expression on mesangial cells was higher by the addition of CM of MNC in MsPGN than in Control, which was prevented by anti-TGF beta antibody or dimethylthiourea. ICAM-1 expression was not different among groups. Thymidine incorporation by adding CM of MNC was lower in MsPGN and PAN, which was mitigated by anti-TGF beta and anti-TNF alpha. Thus the ratio of the expression of ICAM-1 to thymidine incorporation was higher in MsPGN and PAN. CONCLUSION: The expression of beta 1 integrin and ICAM-1, and the thymidine incorporation on mesangial cells are directly regulated by MNC, maybe through factors such as TGF beta or TNF alpha. In MsPGN, the overexpression of beta 1 integrin induced by MNC through TGF beta or hydroxyl radical on day 3 may be related to the pathogenesis.
Animals↗
Degradation products of streptozotocin do not induce hyperglycemia in rats.
The present study was conducted to determine whether degradation products of streptozotocin formed under various conditions induce hyperglycemia in rats. Streptozotocin is completely degraded in pH 7.4 buffer in 4 hr and even more rapidly in plasma. Streptozotocin degradation products resulting from incubation in pH 7.4 phosphate buffer or in plasma were not diabetogenic in rats.
Animals↗
Cloning of cDNA encoding rat phospholipase C-beta 4, a new member of the phospholipase C.
Phospholipase C-beta 4(PLC-beta 4), a new member of phospholipase C isozyme, was purified from bovine cerebellum. The cDNA encoding rat PLC-beta 4 has been cloned from a cDNA library prepared from rat brain. The predicted open reading frame encodes a protein of 1,176 amino acids with a calculated molecular weight of 134,552. The deduced amino acid sequence exhibits 39, 36, and 36% identity with the sequences of rat PLC-beta 1, human PLC-beta 2, and rat PLC-beta 3, respectively. The amino acid sequence of PLC-beta 4, especially, shows higher identity (50%) with norpA PLC sequence from Drosophila melanogaster than those of other PLC-beta subtypes, suggesting that the PLC-beta 4 might be a mammalian PLC equivalent of norpA PLC implicated in photosignal transduction in Drosophila.
Amino Acid Sequence↗
Maximal inspiratory pressure. Learning effect and test-retest reliability in patients with chronic obstructive pulmonary disease.
Reliability of the maximal inspiratory pressure (Pimax) was examined by measuring Pimax once a week for 4 weeks in 91 patients with chronic obstructive pulmonary disease using an aneroid pressure gauge. Five Pimax trials were conducted at each test. From the first to the fourth test, the Pimax increased by a mean of 9 cm H2O (SD = 10). From the third to the fourth test, Pimax increased by a mean of 2 cm H2O and performance appeared to be plateauing. The test-retest reliability coefficient was r = 0.97 for Pimax measured at the third and fourth test session. The 95 percent confidence interval for the absolute difference in Pimax at the third and fourth test was 3 to 5 cm H2O. We conclude that performance of Pimax improves with practice in naive COPD patients and Pimax is reliable when measured with an aneroid gauge by experienced data collectors if patients are given sufficient practice.
Aged↗
Effects of Na+ and Ca2+ concentration in cardioplegic and reperfusion solutions on the intracellular Ca2+ of cardiac muscle cells.
The removal of Ca2+ from the cardioplegic solutions could cause the danger of inducing a "calcium paradox" during reperfusion. Since intracellular Ca2+ activities are coupled to Na+ activities via Na(+)-Ca2+ exchange, an increase in intracellular Na+ activities during the cardioplegia could cause an abrupt Ca2+ influx when reperfused. To study the effects of Na+ and Ca2+ concentrations in cardioplegic solutions on intracellular Ca2+ activities during the cardioplegia and subsequent recovery period, the membrane potential and intracellular Na+ and Ca2+ activities of guinea pig ventricular papillary were measured. 1) A cardioplegia with low Ca2+ cardioplegic solution significantly decreased the overshoot and duration of the first action potential after cardioplegia, but the changes in action potential configuration were minimized after a cardioplegia with Ca2+ concentration adjusted according to the Na(+)-Ca2+ exchange mechanism. 2) Intracellular Na+ activity was continuously decreased during the cardioplegia, and the intracellular Na+ activity 20 minutes after cardioplegia was the highest with low Ca2+ cardioplegic solution. 3) Intracellular Na+ and Ca2+ activities were continuously decreased during the cardioplegia with Ca2+ concentration adjusted according to the Na(+)-Ca2+ exchange mechanism. 4) During a reperfusion of Tyrode solution after cardioplegia intracellular Na+ and Ca2+ activities were increased. Intracellular Ca2+ activity was increased more rapidly than intracellular Na+ activity. 5) The rate of increase in intracellular Ca2+ activity with reperfusion of Tyrode solution was dependent upon intracellular Na+ activity during cardioplegia, in such a way that the higher the intracellular Na+ activity was, the faster the intracellular Ca2+ activity increased. These data suggest that Na(+)-Ca2+ exchange mechanism may play an important role in the regulation of intracellular Ca2+ activity during recovery after cardioplegia.
Animals↗
Crystal structure of papain-E64-c complex. Binding diversity of E64-c to papain S2 and S3 subsites.
In order to investigate the binding mode of E64-c (a synthetic cysteine proteinase inhibitor) to papain at the atomic level, the crystal structure of the complex was analysed by X-ray diffraction at 1.9 A (1 A is expressed in SI units as 0.1 nm) resolution. The crystal has a space group P2(1)2(1)2(1) with a = 43.37, b = 102.34 and c = 49.95 A. A total of 21,135 observed reflections were collected from the same crystal, and 14811 unique reflections of up to 1.9 A resolution [Fo > 3 sigma(Fo)] were used for the structure solution and refinement. The papain structure was determined by means of the molecular replacement method, and then the inhibitor was observed on a (2 magnitude of Fo-magnitude of Fc) difference Fourier map. The complex structure was finally refined to R = 19.4% including 207 solvent molecules. Although this complex crystal (Form II) was polymorphous as compared with the previously analysed one (Form I), the binding modes of leucine and isoamylamide moieties of E64-c were significantly different from each other. By the calculation of accessible surface area for each complex atom, these two different binding modes were both shown to be tight enough to prevent the access of solvent molecules to the papain active site. With respect to the E64-c-papain binding mode, molecular-dynamics simulations proposed two kinds of stationary states which were derived from the crystal structures of Forms I and II. One of these, which corresponds to the binding mode simulated from Form I, was essentially the same as that observed in the crystal structure, and the other was somewhat different from the crystal structure of Form II, especially with respect to the binding of the isoamylamide moiety with the papain S subsites. The substrate specificity for the papain active site is discussed on the basis of the present results.
Amino Acid Sequence↗