Evidence for a role of VLA integrins in lymphocyte-human gingival fibroblast adherence.
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Cell Adhesion Molecules↗
Biomedical subjects
H Okada
Publications and source records attributed to H Okada.
A solid-phase assay for the detection of anti-sperm antibodies.
PROBLEM: ELISA is an ideal assay method for a large-scale screening of anti-sperm antibodies among a large number of infertile males. However, conventional ELISA with whole spermatozoa needs time-consuming steps of centrifugation. METHOD: A solid-phase assay used for detecting anti-sperm antibodies was established. This assay is suitable not only for detecting circulating anti-sperm antibodies of IgG, IgM, and IgA subclass simultaneously but also for screening hybridomas secreting anti-sperm monoclonal antibodies (mAbs). The microtiter plates, on which solubilized sperm antigens are fixed, can be stored at -80 degrees C for up to six months without losing reactivity with anti-sperm antibodies. RESULTS: Using this assay, 53 sera (13 were proven positive and 40 were proven negative for sperm agglutination antibody) were tested. Although the false-negative rate was 0%, the false-positive rate was 32%. One thousand one hundred sixty-five supernatants from hybridomas constructed with splenocytes of mice who were hyperimmunized with human sperm and nonsecreting myeloma cells were tested by this solid-phase assay and two anti-sperm mAb secreting clones were selected and established. CONCLUSIONS: It is recommended that for research work this assay could be used for the first screening of the hybridoma secreting anti-sperm mAb, and for clinical use this assay might be suitable for the first screening of sera of infertile patients. However, conventional bioassays should follow to confirm the biological meaning of the positivity.
Animals↗
[Clinical study of patients with deep-seated fungal infection associated with hematological diseases].
An assessment has been made regarding usefulness of measuring beta-D-glucan (beta-glucan) as fungal serodiagnosis in 50 cases of fungal infection with hematological diseases. Further, an assessment has been made regarding relation between hematological findings and therapeutic effect by administering miconazole, an antifungal agent (MCZ: Florid, clinically to the subjects. Positivity of beta-glucan (beta-glucan > or = 10 pg/ml) was observed in 54.5% (24/44), and the effective rate of MCZ in the positive cases was 75.0% (18/24). In the cases in whom fungus was detected, beta-glucan-positive rate was 50.0% (8/16), and MCZ-effective rate in beta-glucan-positive cases was 62.5% (5/8). The total effective rate of MCZ was 80% (40/50). Side effects were observed in 3 cases, but continual administration of MCZ was possible in all of the 3 cases. By the assessment regarding the relation between hematological findings and therapeutic effect of MCZ, it was found that the effective rates in the cases who underwent a transition with neutrophil and lymphocyte counts less than 500/microliters during the period of MCZ administration were 64.7% (11/17) and 50% (5/10), respectively, and large effects were observed in the cases who underwent a transition with the neutrophil and lymphocyte counts more than 500/microliters was 86.7% (19/22) and 91.7% (22/24), respectively. These results suggested that lymphocytes rather than neutrophils had an important role in the morbidity of fungal infection. It was noteworthy that MCZ was effective for the treatment of deep seated mycosis and significant effective rate was obtained in the group of patients who had neutrophils and lymphocytes less than 500/microliters.
Adolescent↗
Nasal carriers of Staphylococcus aureus among healthy individuals.
Staphylococcus aureus and Staphylococcus spp. were isolated from healthy students and their drug resistance was investigated. S. aureus was isolated from 17 of 70 persons (24.3%). Fourteen strains of 22 isolates of S. aureus were ampicillin-resistant. Two strains each were cefmetazole-resistant and gentamicin-resistant. None of the strains was found to be methicillin-resistant. Compared with the strains isolated from the hospital ward environment, S. aureus in healthy individuals was relatively sensitive to antibiotics.
Adult↗
Nylon oligomer degradation gene, nylC, on plasmid pOAD2 from a Flavobacterium strain encodes endo-type 6-aminohexanoate oligomer hydrolase: purification and characterization of the nylC gene product.
A new type of nylon oligomer degradation enzyme (EIII) was purified from an Escherichia coli clone harboring the EIII gene (nylC). This enzyme hydrolyzed the linear trimer, tetramer, and pentamer of 6-aminohexanoate by an endo-type reaction, and this specificity is different from that of the EI (nylA gene product) and EII (nylB gene product). Amino acid sequencing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified EIII demonstrated that the enzyme is made of two polypeptide chains arising from an internal cleavage between amino acid residues 266 and 267.
Amidohydrolases↗
Steroid-responsive renal insufficiency due to idiopathic granulomatous tubulointerstitial nephritis.
A 26-year-old male was admitted to our hospital having suffered from subfever, sterile pyuria and renal insufficiency for 1 year. There had been no apparent hematuria/proteinuria, and evidence of infection, allergy or intoxication was not demonstrated. Open renal biopsy revealed severe tubulointerstitial changes with granuloma accompanied by neither caseous necrosis nor giant cells. Infiltrating cells mostly consisted of CD4 and CD8 T cells, and some proximal tubular cells presented MHC class II antigen. Following the negative culture of biopsied specimen for acid-fast bacilli, diagnosis of idiopathic granulomatous tubulointerstitial nephritis was made, and steroid therapy was started. Two months later, pyuria disappeared and renal function improved significantly, proving the effectiveness of steroid on idiopathic granulomatous tubulointerstitial nephritis.
Adult↗
Phosphatidylinositol metabolism in hypertrophic rat heart.
The accumulation of inositol 1,4,5-trisphosphate (IP3) after hormonal stimulation has a physiological role, possibly by alteration of Ca2+ levels in cardiac myocyte. However, this accumulation has not been studied under pathophysiological conditions. In this report, we examine phosphatidylinositol metabolism during cellular response to norepinephrine in pressure-overloaded hypertrophic rat heart. After stimulation with norepinephrine, the accumulations of IP3 and diacylglyceride significantly increased in isolated myocytes from stroke-prone spontaneously hypertensive rat (SHRSP) heart, indicating phosphatidylinositol-specific phospholipase C activity increased in SHRSP heart cells. Protein kinase C activity was also enhanced in SHRSP, with a marked increase in particulate activity. We determined the intracellular calcium concentration and found it to be higher in SHRSP than in Wistar-Kyoto (WKY) rats at 30-40 weeks of age. Ca2+ influx was also elevated in SHRSP stimulated by norepinephrine. In SHRSP heart, cytosolic Ca2+ concentration may rise quickly in response to some stimuli, such as alpha 1-adrenergic stimulation, which is shown to be one of the pathways that increases cytosolic Ca2+ levels in hypertrophied rat heart. These data suggest that a part of the phosphatidylinositol-turnover pathway, such as the phosphatidylinositol 4,5-bisphosphate-IP3-Ca2+ pathway or the diacylglyceride-protein kinase C pathway, may play an important role in the development of hypertrophy in SHRSP heart.
Animals↗
Expression of 20 KD homologous restriction factor of complement on myocardial cells: an immunohistochemical study using acetone-fixed and paraffin-embedded tissues.
Expression of 20 KD homologous restriction factor of complement (HRF20) (CD59) in normal human heart was immunohistochemically examined with monoclonal antibody (MAb) 1F5. HRF20 was clearly demonstrated on the cell surface membrane and intercalated discs of myocardial cells throughout the ventricular walls. Therefore, this factor may protect normal cardiomyocytes from complement deposition, which has been demonstrated on infarcted cardiomyocytes. Expression of HRF20 was also observed on endothelial cells of the endocardium and on blood vessels, including arteries, capillaries, and veins, and on the Schwann-cell sheath of peripheral nerve fibers. In the present study, we found that the epitopes of HRF20 were well preserved for immunohistochemistry even in acetone-fixed and paraffin-embedded tissues, as well as in frozen tissues that have been conventionally used for HRF20. This method using paraffin sections allows for easier staining and greater accuracy in histological examination.
Acetone↗
[The relationship between cardiac systolic and diastolic function and the changes in cellular membrane signal transduction system and extracellular matrix].
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Animals↗
Expression of the cellulase (FI-CMCase) gene of Aspergillus aculeatus in Escherichia coli.
FI-CMCase cDNA of Aspergillus aculeatus was expressed in Escherichia coli by using the tac promoter of E. coli. Transformants of E. coli harboring a plasmid pHEM06 containing mature form FI-CMCase cDNA produced FI-CMCase in the cytoplasm of the cells. The enzyme from E. coli cells was purified to yield 56% and it was immunological identical to that of FI-CMCase purified from A. aculeatus.
Amino Acid Sequence↗
Pharmacokinetics and tolerability of chloramphenicol in budgerigars (Melopsittacus undulatus).
Pharmacokinetics and tolerability of chloramphenicol (CP) were evaluated in budgerigars. Following intramuscular administration of CP at 100 and 200 mg/kg of body weight, serum peak concentrations of 35.3 and 90.7 micrograms/ml, respectively, were obtained 0.25 hr following injection, and these values declined with a terminal half-life of 2.5 and 2.7 hr, respectively. Based on these results, an intramuscular dosage regimen of 100 mg/kg of CP 3 times a day or of 200 mg/kg twice daily was recommended as effective against many of the common bacterial infections in budgerigars. Red blood cell count, packed cell volume, hemoglobin, total plasma protein, serum AST, ALT, LDH and CK values were determined after intramuscular injection of either CP or physiological saline 2 or 3 times a day for 5 days. The birds were weighed before and after treatment. Tissue samples from various organs were examined histologically. The most prominent adverse effect was muscular damage at the injection site. A dosage regimen of 200 mg/kg twice daily for 5 days was considered to be safe.
Alanine Transaminase↗
First incidence of multilocular echinococcosis in a race horse in Japan.
Nodular hepatic lesions caused by larval Echinococcus multilocularis were found in a four-year-old female thoroughbred race horse born and raised in Hokkaido. Before detection of the infection, the infected animal was transported around Honshu for racing. The present disclosure of hydatidosis in a race horse raises concern over the potential danger of spreading the infection throughout Japan by horses possibly infected in Hokkaido.
Adrenal Glands↗
Immunohistochemical localization of 17 alpha-hydroxylase/C17-20 lyase and aromatase cytochrome P-450 in polycystic human ovaries.
Immunohistochemical localization of 17 alpha-hydroxylase/C17-20 lyase (P-450(17 alpha,lyase)) and aromatase cytochrome P-450 (P-450arom) in polycystic ovary (PCO) syndrome was studied using specific polyclonal antibodies which had been raised against the corresponding enzymes. In the majority of follicles that were atretic and smaller than 7 mm in diameter, theca interna cells showed high P-450(17 alpha,lyase) immunoreaction, while small numbers of granulosa cells showed little P-450arom immunoreaction. In some atretic follicles that were larger than 11 mm in diameter, the hyperplastic theca interna cell layer showed high immunoreaction to P-450(17 alpha,lyase), while the poorly proliferated granulosa cell layer showed a mixture of weak and negative immunoreaction to P-450arom. No immunoreaction to P-450(17 alpha,lyase) or P-450arom was recognized in PCO stroma. These findings suggest that the theca interna cells and the granulosa cells from PCOs show abnormal steroidogenic function, while the localization of P-450(17 alpha,lyase) and P-450arom in PCOs was essentially identical to that in the normal ovary. Theca interna cells in PCO atretic follicles are the main site of excess androgen production.
Adult↗
[Ultrastructure of immotile spermatozoa obtained from infertile male patients].
We sometimes experienced infertile patients whose sperms had no motility but were not stained by Eosin Y. In this paper we report five cases of so-called "immotile spermatozoa". The ultrastructure of sperm tails was examined by transmission electrone microscope (TEM). These cases were selected from the out-patient population who attended infertility clinic of our department. The semen analyses showed that all the cases had sperm motility below 1% and more than 90% of the spermatozoa were proven alive. Family history revealed that one case had an infertile sibling. None of them had situs inversus, bronchiectasis and chronic sinusitis which are classic trias of Kartagener's syndrome. They had no symptoms of upper respiratory tract infection which was caused by the abnormality in the flagella of the respiratory tract. The TEM pictures of sperm tails showed partial deletion of inner dynein arms in two cases, lack of central microtubular doublets (so-called 9 + 0) in two cases and disarrangement of microtubular doublets in one case. For the treatment of these cases there is no effective means but AID. However, the rapid progress of IVF-ET techniques and a report that the spermatozoa from Kartagener's syndrome had showed penetration into eggs encouraged us to think the micromanipulation of spermatozoa with IVF-ET as a hopeful option of the treatment in the near future.
Adult↗
[Subpopulation of leucocytes in the semen from fertile and infertile men--immunohistochemical and flow-cytometric analysis].
It is hard to differentiate leucocytes from immature germ cells among many round cells in ejaculated semen. Recently monoclonal antibodies (MoAbs) which recognize surface antigen of various subtypes of leucocytes are available. In this study we applied a panel of MoAbs for leucocyte subpopulations (granulocytes, T-lymphocytes, B-lymphocytes, Monocytes/Macrophages and pan-leucocytes). Semen smears from 64 mild oligozoospermic patients and 5 fertile volunteers were stained by immunohistological technique (avidin-biotin-immunoperoxidase method). We found highly varying total leucocyte numbers ranging from 1200 to 2.2 x 10(8) per ejaculate. When comparing semen from infertile patients with those from fertile donors, statistically significant higher medians of total leucocytes, total granulocytes and monocytes/macrophages were seen in infertile group. We compared a total number of leucocytes obtained from conventional Papanicolaou staining with that from immunohistochemical staining. Both were almost identical but the former gave a little bit higher value, because of the difficulty in differentiating leucocytes from immature germ cell by Papanicolaou staining. Moreover we performed an experiment to evaluate the applicability of flow-cytometry (FCM) to count leucocytes in semen. Cells in the semen smears were stained with MoAbs recognizing the surface antigen shared by every leucocyte, and percentage of the leucocytes which were thus stained with MoAbs was compared with that determined by FCM. Results from FCM constantly gave lower value than these from immunostaining. This is probably because FCM could not differentiate leucocytes from contaminated non cell substances (e.g. cell debris). From the present study it is not adequate to use FCM for the detection of leucocytes in semen as applied in haematology. Further techniques to remove non-cellular substances should be established for its application in semen analysis.
Antibodies, Monoclonal↗
Antitumor effects of angiogenesis inhibitor TNP-470 in rabbits bearing VX-2 carcinoma by arterial administration of microspheres and oil solution.
The antitumor activities of an angiogenesis inhibitor, (3R,4S,5S,6R)-5- methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-2-butenyl)-oxiranyl]-1- oxaspiro[2,5]oct-6-yl(chloroacetyl) carbamate (TNP-470), administered s.c., i.v. and intra-arterially using a chemoembolization technique, were examined in rabbits bearing VX-2 carcinoma. Repeated s.c. injection of TNP-470 aqueous solution clearly inhibited the tumor growth in proportion to the dose, and improved efficacy was obtained with once a week s.c. administration of microspheres, which provide sustained release of TNP-470. Moreover, the injection of the TNP-470 microspheres into the predominant artery of the tumor via a catheter, chemoembolization, resulted in dramatic regression of the tumor. This antitumor effect was enhanced by coadministration of doxorubicin hydrochloride aqueous solution. Arterial administration of TNP-470 oil solution provided more persistent tumor growth inhibition due to the prolonged release and targeting of the angiogenesis inhibitor to the tumor.
Animals↗
[An investigation of the collagen in cardiomyopathic hamsters].
Extensive fibrosis is remarkable in cardiomyopathy. The purpose of this study is to characterize the collagen, one of the chief elements of the extracellular matrix of hereditary cardiomyopathic hamsters, and to examine the participation of the collagen in the occurrence and the progression of the cardiomyopathy. BIO53.58 (5, 11, 22 weeks) was used as the model of the dilated cardiomyopathy and BIO14.6 (20, 30 weeks) was used as the model of the hypertrophic cardiomyopathy F1b was used as the control. The collagen content was almost constant at any age in F1b, but increased with aging in BIO53.58 and BIO14.6. Type III collagen increased significantly in BIO53.58 at 11 weeks and BIO14.6 at 20 weeks. Type V collagen decreased significantly in BIO14.6 at 30 weeks. Acetic acid solubility of collagen decreased in BIO53.58 and BIO14.6 with the progression of the fibrosis, but not in F1b. Reducible crosslinks showed the tendency to decrease in BIO53.58 progressively. Histologically thick collagen fiber increased in BIO53.58 and BIO14.6. These findings indicate that in the early phase of the cardiomyopathy the extracellular matrix of the myocardium has characteristics that of the immature tissues which are rich in type III collagen. In later phase, the matrix resembles that of hard tissues whose collagen is mainly of type I collagen and have low solubility. It is considered that the increase of the thick collagen fiber combined firmly in heart may affect the diastolic and the systolic function in addition to the loss of the cardiac muscle fiber by the degeneration and the necrosis.
Animals↗
Thrombin generates monocyte chemotactic activity from complement factor H.
We have recently found that the complement factor H (H) was the precursor of the major macrophage chemotactic factor in the delayed-type hypersensitivity (DTH) reaction site in the skin and was converted to the factor by an unidentified trypsin-like protease in plasma. Thrombin and plasmin are also present in the site, and we, therefore, examined the possibility that these proteases converted H to be monocyte chemotactic. Intact H caused no monocyte migration, although it was able to do so after incubation with thrombin, but not with plasmin. The activity was chemotactic rather than chemokinetic and was absorbed by an anti-H IgG-conjugated column. The generation of monocyte chemotactic activity from H was dependent on incubation time with thrombin and also the protease activity of thrombin, and the activity was seen at concentrations of H lower than 10(-8) M. The inhibitory activity of H for C3b-Bb was not affected by incubation with thrombin or plasmin. Incubation of H with thrombin, but not with plasmin, generated a hydrophobic molecule, in a time-dependent manner, which had monocyte chemotactic activity. These results show that H becomes a monocyte chemotactic factor due to cleavage by thrombin, which converts H to a more hydrophobic molecule and also suggest that thrombin-treated H induces monocyte migration in the DTH reaction site.
Chemotaxis, Leukocyte↗