[Transmission of HTLV-I].
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Biomedical subjects
Publications and source records attributed to Y Ando.
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We measured appearance rates of RNA-rich reticulocytes in 8 patients undergoing allogeneic bone marrow transplantation, using a Sysmex R-1000 reticulocyte counter which utilizes laser flow cytometry. The changes in proportion of RNA-rich reticulocytes (high fluorescence ratio: HFR) and maturation index (MI: HFR + middle fluorescence ratio/low fluorescence ratio) were compared with those of WBC, neutrophil or reticulocyte counts. Engraftment was defined as an HFR of greater than or equal to 5% or a MI of greater than or equal to 15%. Engraftment was confirmed significantly earlier by HFR (13.5 +/- 2.4 days) and MI (13.1 +/- 2.9 days) than by neutrophils (17.1 +/- 3.2 days) or reticulocytes (20.4 +/- 6.2 days). The maturation of reticulocytes would be a useful indicator for engraftment or recovery from marrow aplasia in cases of bone marrow transplantation.
HHV-6 which was detected in peripheral blood lymphocytes of lymphocytic patients is now said to be an agent causing exanthema subitum. We investigated the titer of HHV-6 antibody in 100 primigravida and 30 patients with spontaneous abortion by means of the indirect immunofluorescence method (IF method). The positive incidence in which the titer of HHV-6 antibody was more than 10 times in 100 primigravida was 82%. That of HHV-6 IgM antibody was 3% in those who had no complication during pregnancy. This supposes that there may be some women who were infected or were reactivated while they were pregnant. The positive incidence in 30 spontaneous abortion patients was 90%. 3 of them had IgM antibody to HHV-6, and HHV-6 antigen was detected on the epitherium of the chorionic villi by the IF method with HHV-6 monoclonal antibody in two of them. This suggests that the infection or reactivation of HHV-6 may be one of the causes of spontaneous abortions.
We analyzed the RNA in platelets by fluorescence flow cytometry after staining with thiazole orange(TO) in whole blood samples from hematologically normal subjects and patients with thrombocytopenia. The percentage of TO-positive platelets and their mean fluorescence channel number in 32 control subjects were 6.2 +/- 2.5% (mean +/- SD) and 6.9 +/- 0.7, respectively. In 11 patients with idiopathic thrombocytopenic purpura, the percentage of fluorescently labeled platelets was significantly elevated (p less than 0.05) to 21.5 +/- 14.3%. By contrast, the proportion of positively stained platelets in 14 patients with thrombocytopenia due to impaired platelet production did not significantly differ from that of the controls, whereas the absolute counts of TO-positive platelets were significantly lowered (p less than 0.05). In both patient groups, the mean fluorescence channel numbers of TO-positive platelets were significantly elevated to 16.1 +/- 16.8 and 6.9 +/- 0.7, respectively (p less than 0.05, 0.005). We conclude that flow cytometric analysis of platelets after staining with TO provides information on the thrombopoietic activity in thrombocytopenic disorders. The main advantages of this method for clinical use are its simplicity and the rapidity.
Using a methylcellulose culture system, we studied the effects of recombinant human interleukin-3 (IL-3), recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF), and recombinant human granulocyte colony-stimulating factor (G-CSF) on the growth of myeloid progenitor cells (CFU-C) from an adult patient with congenital neutropenia. The moderate clinical course and the maturation arrest at blast-promyelocyte stage in the marrow differentiated this patient from those described as having Kostmann-type congenital neutropenia. CFU-C growth in bone marrow cells from the patient responded to IL-3 normally in a dose-dependent manner. GM-CSF stimulated only macrophage colony formation in a dose-dependent manner comparable to that in normal subjects. Neither GM-CSF nor G-CSF stimulated any significant granulocyte colony formation. This evidence suggests that the hematopoietic progenitor cells in this patient had the potential for developing CFU-C with IL-3, and that the neutropenia in this patient could be a result of an intrinsic defect in myelopoiesis along a granulocytic pathway responsive to GM-CSF or G-CSF.
In patients with cervical lymph node swelling, prompt differential diagnosis of tuberculosis leads to an appropriate treatment. A method based on DNA amplification and hybridization for the rapid detection of Mycobacterium tuberculosis was used for demonstrating its specific DNA in cervical lymph nodes biopsied from seven patients. The DNA specific for Mycobacterium tuberculosis was detected in 5 patients. Four of them were histologically diagnosed as tuberculous lymphadenitis. In another case with necrotizing lesions but without granulomatous reactions, culture method subsequently revealed a positive result. In other 2 cases, the diagnosis of tuberculosis was promptly excluded by both this method and conventional methods (histologic diagnosis/direct microscopy). The rapid detection of Mycobacterium tuberculosis by amplification of mycobacterial DNA is helpful particularly in differential diagnosis of cervical lymphadenopathy.
From 1975 to 1990, we treated 118 patients with urinary epithelial cancer, including 100 with primary bladder cancer, 13 with primary upper urinary tract cancer, and 5 with both diseases. Thirty-five patients with primary bladder cancer underwent total cystectomy. Upper urinary tract urothelial cancer developed in 4 patients (4.0%) and was detected only after cystectomy. Three patients had multiple bladder tumors before cystectomy and recurrent tumors under long-term bladder-preserving treatment. The other patient had had cystectomy for the primary bladder lesion. Our present policy is to perform urinary cytology once a month and intravenous urography once a year in patients with bladder cancer for early detection of secondary upper urinary tract cancer.
Using Cox's proportional hazard model, we performed a multivariate analysis of survival data from 126 patients who underwent curative resection for colorectal cancer from 1971 to 1985. Flow cytometric DNA measurements were carried out using paraffin wax-embedded tissue blocks. Fifty-four per cent of the cases were found to be aneuploid and 46% were diploid. The depth of invasion of the tumor, nodal status, liver metastasis, peritoneal dissemination, DNA ploidy pattern, histology, macroscopic classification of the tumor, age, sex and site were tested in a survival analysis. The initial seven factors were found to be the significant prognostic variables in a univariate analysis. A multivariate analysis shows liver metastasis, DNA ploidy pattern and peritoneal dissemination as being the significant discriminants of survival (P = 0.0001, 0.0022, 0.0119, in this order). Therefore, nuclear DNA ploidy pattern in colorectal cancer is considered to be an independent prognostic factor.
Laboratory and clinical studies on cefprozil (CFPZ, BMY-28100), a new cephem antibiotic, were carried out in the field of pediatrics. The results obtained are summarized as follows: 1. Serum concentrations, urinary concentrations and urinary recovery rates of CFPZ were determined upon oral administration of CFPZ after meal at doses of 4 mg/kg granules in a case, 7.5 mg/kg granules in 2 cases and 15 mg/kg granules in one. Peak serum levels of CFPZ were obtained at an hour in 3 cases and at 2 hours in 1 case after administration of the drug with a range of 2.7-8.6 micrograms/ml with half-lives of 0.69-0.95 hours. Urinary recovery rates in the first 6 hours after administration ranged from 59.4-71.3%. 2. MICs of CFPZ against 36 clinical isolates (Staphylococcus aureus 4 strains, Streptococcus pneumoniae 5, Streptococcus pyogenes 5, Escherichia coli 5, Haemophilus influenzae 12, Haemophilus parainfluenzae 4, and Branhamella catarrhalis 1) were compared with those of cefaclor (CCL) and ampicillin (ABPC). The antibacterial activity of CFPZ was superior to those of CCL against Gram-positive cocci, and to those of ABPC against E. coli, and was equal to those of CCL and inferior to those of ABPC against H. influenzae. 3. Thirty-seven pediatric patients with acute infectious diseases (pharyngitis/tonsillitis 17, bronchitis 7, pneumonia 3, skin and soft tissue infection 2, and urinary tract infection 8) were treated with CFPZ at daily doses of 10-47 mg/kg t.i.d. as a rule. The efficacy rates were 100% clinically and 56% bacteriologically. 4. Side effects or abnormal laboratory test values were not observed except for an increased platelet count in 1 case and elevated GOT, GPT values in 2 cases.
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In patients with malignant blood disease, infection is the most serious complication. The prompt differential diagnosis of tuberculosis leads to an appropriate treatment. A method based on DNA amplification and hybridization for the rapid detection of Mycobacterium tuberculosis was used to test 10 clinical specimens (sputum, gastric aspirate and pleural effusion) from blood disease patients in whom tuberculosis was suspected. Mycobacterium tuberculosis DNA was detected in 3 specimens, including one which was negative on standard microbiological criteria (microscopy and/or culture). The other 7 specimens with fever or abnormal shadow on a chest X-ray were negative by both our method and the Standard microbiological criteria. Rapid diagnosis of tuberculosis by amplification of mycobacterial DNA in cases of blood disease in clinically useful.
The autonomic nervous system innervates every visceral and peripheral organs in the body. It has as complex a neural organization in the brain, spinal cord, and periphery as the somatic nervous system, but remains largely involuntary or automatic. Accurate diagnosis is essential for proper management of autonomic failure but in attempting to classify autonomic disease there is a philosophical point to be borne in mind. Since it is very difficult to comprehend and evaluate systematically the aspect of autonomic dysfunction in each organ in various diseases, little works have been performed to classify autonomic diseases. We introduce several comprehensible works of classification of autonomic failure. We also presented some typical autonomic diseases according to those classifications in this paper.
We purified a new EF-hand type calcium binding protein from chicken gizzard smooth muscle, tentatively named calgizzarin (Todoroki, H., et al. J. Biol. Chem. (1991) in press. Based on the internal peptide sequence of calgizzarin, we isolated and sequenced a cDNA clone coding for calgizzarin from a rabbit lung cDNA library. This clone (pCALG) has 309 nucleotides of open reading frame including termination codon TGA, 621 nucleotides of the 5' leader and 186 nucleotides of the 3' noncoding region. The polypeptides deduced from the open reading frame were consisted of 102 amino acid residues with a molecular weight of 11,429. Computer aided homology analysis revealed that calgizzarin exhibits a 43.2% homology to S-100 alpha, 38.6% to S-100 beta and 40.0% to annexin II light chain, p10. By Northern blot analysis, with pCALG, a band of 1.1 kbp was detected in rabbit lung, suggesting pCALG contains nearly full length of mRNA.
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Although the possible involvement of superoxide radical and its metabolite(s) in the pathogenesis of various types of edema have been suggested, direct evidence supporting this concept is lacking. Since intravenously administered Cu2+Zn2(+)-type superoxide dismutase (SOD) rapidly disappeared from the circulation with a half-life of 4 min, the enzyme could not be used to test whether superoxide radicals played a critical role in the modulation of vascular permeability. We previously synthesized a SOD derivative (SM-SOD) by linking poly(styrene co-maleic acid butyl ester) (SM) to the enzyme (Ogino, T., Inoue, M., Ando, Y., Awai, M., Maeda, H. and Morino Y. (1988) Int. J. Pept. Protein Res. 32, 1583-1588); SM-SOD circulates bound to albumin with a half-life of 6 h. To test whether superoxide radicals play an important role in the regulation of vascular permeability, the effect of SM-SOD on experimental paw edema was studied in the rat. Subcutaneous injections of carrageenin to the paw rapidly induced local edema by increasing vascular permeability. Intravenous administration of SM-SOD markedly inhibited the carrageenin-induced increase in vascular permeability and suppressed the development of paw edema. In contrast, the same dose of SOD showed no such inhibitory effect. These results suggest that superoxide radical and/or its metabolite(s) might play a critical role in the pathogenesis of carrageenin-induced vasogenic edema.
Covalently cross-linked multimers of lipocortin I are shown to be present in human epidermoid carcinoma A431 cells treated with epidermal growth factor or the calcium ionophore A23187. This intracellular cross-linking of lipocortin I is suggested to be mediated by the action of tissue transglutaminase, a Ca2(+)-dependent protein cross-linking enzyme. Cross-linking of lipocortin I competes with proteolytic digestion of the protein, and pretreatment of the cells with inhibitors for calpain (Ca2(+)-dependent intracellular protease) markedly enhanced the cross-linking of lipocortin I. Cross-linked lipocortin I is shown to be present in the soluble fraction of A431 cells as well as in the particulate fraction; a 34-kDa fragment of lipocortin I was solubilized successfully by plasmin digestion of the latter fraction. Immunofluorescence microscopy using specific antilipocortin-I antibody showed that cross-linked lipocortin I forms an envelope-like structure, which is not extracted with [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) or Triton X-100. In vitro incubation of purified lipocortin I with tissue transglutaminase resulted in the formation of covalently cross-linked lipocortin I dimer, tetramer, and so on. Amine incorporation and cross-linking studies using lipocortin I and its N-terminal truncated derivatives indicated that the cross-linking site is localized within the plasmin-susceptible N-terminal 29 amino acids of lipocortin I. The cross-linking of lipocortin I is shown to be accelerated more than 10 times by the addition of phosphatidylserine vesicles, on which lipocortin I molecules are most likely aligned in a conformation suitable for cross-linking. Collectively, these findings suggest that an increase of intracellular calcium concentration results in the attachment of lipocortin I onto the plasma membrane phospholipids through the C-terminal domain of the molecule where the membrane-bound lipocortin I is cross-linked by the action of tissue transglutaminase through the N-terminal domain.