Serologic tests for Chlamydia pneumoniae.
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Adult↗
Biomedical subjects
T Hagiwara
Publications and source records attributed to T Hagiwara.
[Effect of storage conditions on the infectivity of Chlamydia pneumonia].
In order to facilitate the isolation of C. pneumoniae, strain similar TWAR, with high frequency, we investigated the effect of various factors on the infectivity of Chlamydia using two laboratory strains, C. pneumoniae TWAR and C. trachomatis serovar D. The factors tested were the effects of different temperatures for storage conditions, saliva from healthy person, storage media for Chlamydia, and the frequency of freezing and thawing. Chlamydial suspension was prepared in the two media, SPG (sucrose-phosphate-glutamate buffer pH 7.5), and CT-GM (culture medium for Chlamydia which contains 1 micrograms/ml cycloheximide and 0.04% glucose). Chlamydial suspension was allowed to stand in each of four different thermal conditions: 37 degrees C, room temperature (25 degrees C), 4 degrees C, 0 degrees C and -75 degrees C for 1, 3, 6, 24, 48, 72, 96, 120 and 144 hours. For storage at -75 degrees C, one of three groups of glass vial tubes containing Chlamydia was covered with an "airmat" to prevent the rapid freezing of Chlamydia. The effect of various factors on the infectivity was assayed by inoculation of the suspension on HeLa 229 cell monolayers. Results showed that the infectivity rapidly decreased at 37 degrees C and room temperature, while at 4 degrees C, 0 degrees C and -75 degrees C, relatively high infectivity was maintained and contained until days 4 to 6. This decreasing pattern was similar to among the media used. We were not able to find any differences in the infectivity among the samples with or without the "airmat".(ABSTRACT TRUNCATED AT 250 WORDS)
Chlamydia↗
Interstitial pneumonitis possibly due to mitoxantrone.
A 41-year-old patient with chronic myelogenous leukemia in the accelerated phase was treated with mitoxantrone. She developed pyrexia 7 days after receiving the third administration of mitoxantrone. After 3 more days, she experienced dry cough and dyspnea. Bilateral fine crackles were audible, but no signs of heart failure were found. A chest X-ray film revealed diffuse reticulogranular infiltrates bilaterally. An increase in the prednisolone dosage led to an improvement. Specimens of the bronchoalveolar lavage revealed an increase in CD4-/CD8- lymphocytes. The peripheral lymphocytes also expressed neither CD4 nor CD8. Specimens of a transbronchial lung biopsy disclosed thickening of the alveolar wall with infiltration of lymphoid cells.
Adult↗
[Separation of glycoproteins from cyst fluids of human ovarian cystomas in benign and their some chemical and serological properties].
Perchloric acid-soluble fractions (PASFs 1-3) which were obtained from cyst fluids of two patients [(Nos. 1 (blood group B) and 2 (blood group A))] with ovarian mucinous cystadenoma and one patient (No. 3 (blood group A)) with ovarian dermoid cyst in benign, were identified as glycoproteins by their chemical composition analyses, respectively. Reactivities of PASFs 1-3 against blood group anti-sera and lectins suggest that these PASFs did not contain MN blood group substances, Vgu glycoproteins and T-active glycoproteins. PASF 1 was separated into 11 fractions by Sephacryl S-500 gel filtration. From chemical composition, SDS-PAGE and serological property of the main fraction (Fr. 5), other one fraction (Fr. 7) and mild acid hydrolysates of these two fractions, it is possible to presume that PASF 1 consisted of a large number of AB-active and fucose-rich glycoproteins with high molecular weights and some of these glycoproteins bear either NeuNAc alpha 2-->6GalNAc (sialyl Tn) group or both sialyl Tn and sialylated T groups.
Adult↗
Immunohistochemical analysis of tissue distribution of B and T cells in germfree and conventional chickens.
The distribution of lymphocytes in lymphoid organs of germfree (GF) and conventional (CV) chickens was compared immunohistochemically with several monoclonal antibodies. The lymphoid follicles in the cecal tonsils of GF chickens were more poorly developed than in those of CV chickens, and IgG and IgA positive cells were also not found in GF chickens. The number of both B and T cells in the villous region of the cecal tonsils in GF chickens was less than that in CV chickens. In the spleen of GF chickens, the distribution of B cells was similar, but T cells were distributed more widely in CV chickens than in GF chickens. There was no clear difference in the distribution of lymphoid cells in the bursa of Fabricius and the thymus. These results suggest that the development of gut-associated lymphoid tissues is affected by intestinal flora.
Animals↗
Historical control data of organ weight and gross findings in F344/DuCrj rats and B6C3F1 mice.
Organ weight and gross postmortem findings of control Fischer 344/DuCrj rats and B6C3F1 mice are presented from subchronic, chronic toxicity and carcinogenicity studies that were conducted over a 9 year period at our center (An-Pyo Center). The mean organ weight of the liver, kidney, spleen and the lung were increased associated with age in both species. The most common findings observed at 109 weeks in both species were thymic atrophy, enlargement of the spleen, dilation of the lumen of the uterus and ovarian cysts. Male and female Fischer 344/DuCrj rats commonly exhibited a granular surface of the kidneys, hypertrophy of and/or nodular pituitary glands and subcutaneous tissue masses. Multiple white patches and hypertrophy or atrophy of the testes, atrophy of the seminal vesicles and prostate and nodules in the pancreas were seen frequently in male rats. Malformative nodule in the liver was a common finding in female rats. The most common lesions seen in both male and female B6C3F1 mice were nodules in the liver and lungs and enlargement of the lymph nodes. Nodules on the preputial gland were commonly seen in the males. Sex differences were evident in the incidence of some of the above findings. These historical data will contribute to analyze the organ weight and gross findings at necropsy in long-term toxicity and carcinogenicity studies.
Animals↗
[Serum and urinary neopterin levels in sarcoidosis].
Serum and urinary neopterin levels were determined by high pressure liquid chromatography in 26 patients with pulmonary sarcoidosis and 12 healthy controls. Neopterin levels were significantly higher in sarcoidosis patients than in the controls. Neopterin levels differed from serum angiotensin converting enzyme (ACE) activities and were significantly higher in radiologic stage 1 and 2 than in radiologic stage 0 in patients not receiving prednisolone. No significant correlation was found between neopterin level and serum ACE activity. On the other hand, a significant correlation was found between neopterin level and serum adenosine deaminase activity. We believe that serum and urinary neopterin levels may be a more clinically valuable method of assessing pulmonary sarcoidosis than serum ACE activity.
Adult↗
Specific phosphorylation of the acidic central region of the N-myc protein by casein kinase II.
The central region of the N-myc protein has a characteristic amino acid sequence EDTLSDSDDEDD, which is very similar to those of particular domains of adenovirus E1A, human papilloma virus E7, Simian virus 40 large T, c-myc and L-myc proteins. Domains of these three viral oncoproteins have recently been shown to be specific binding sites for the tumor-suppressor gene retinoblastoma protein. We have noted that the sequence of serine followed by a cluster of acidic amino acids is exactly the same as that of a typical substrate of casein kinase II (CKII). Therefore, we investigated whether these nuclear oncoproteins are phosphorylated by CKII. For this purpose, we fused the beta-galactosidase and N-myc genes including this domain and expressed it in Escherichia coli cells. Several mutant N-myc genes, containing single amino acid substitutions in this domain, were also used to produce fused proteins. Strong phosphorylation by CKII was detected with the fused protein of wild-type N-myc. However, no phosphorylation of beta-galactosidase itself was observed and the phosphorylations of fused mutant proteins were low. Another fused N-myc protein containing most of the C-terminal region downstream of this acidic region was not phosphorylated by CKII. Analysis of phosphorylation sites in synthetic peptides of this acidic region identified the major sites phosphorylated by CKII as Ser261 and Ser263. On two-dimensional tryptic mapping of phosphorylated N-myc proteins, major spots of in vitro-labeled and in-vivo-labeled N-myc proteins were detected in the same positions. These results suggest that two serine residues of the acidic central region of the N-myc protein are phosphorylated by CKII in vivo as well as in vitro. The functional significance of this acidic domain is discussed.
Amino Acid Sequence↗
Logarithmic temperature dependence of resistivity in heavily doped conducting polymers at low temperature.
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Journal Article↗
Expression and function of adhesion molecules on human hematopoietic stem cells: CD34+ LFA-1- cells are more primitive than CD34+ LFA-1+ cells.
Advances in fluorescence-activated cell sorter technology have brought about multicolor analysis of cell phenotypes. To clarify the phenotypes of human hematopoietic stem cells (HSCs), we initially prepared novel antibodies against CD34 and labeled one of them (4A1) with allophycocyanin (APC). With this, we analyzed the phenotypes of CD34+ HSCs and showed that primitive HSCs or CD34+CD33- cells expressed adhesion molecules such as CD43, CD44, CD11a, CD11c, CD18, and leukocyte adhesion molecule (LAM-1). The more primitive hematopoietic cells or CD34+CD38- cells also expressed CD11a and CD18 with an incidence of 20% to 30%. To clarify the role of adhesion molecules in HSCs, we examined the colony forming capacity after long-term culture with allogeneic irradiated stromal layers. Among CD34+CD33- cells, CD18+ cells gave rise to colony-forming cells (CFCs) on stromal layers, but reached a maximum at week 2, after which the number of generated CFCs decreased. On the other hand, CD18- cells generated less CFCs than CD18+ cells at 2 to 3 weeks, but increased after 4 weeks of culture. When CD18 or CD11a antibody was added to a coculture system of CD34+CD33- cells with stromal layers, the number of generated CFCs decreased significantly compared with the no antibody control. Leukocyte function-associated antigen-1 (LFA-1) (CD11a/CD18) was expressed on some populations of hematopoietic cells and contributed to the proliferation by interacting with stromal cells. However, more primitive cells capable of reconstituting hematopoiesis did not express LFA-1. These data provide a rationale for the administration of anti-LFA-1 antibody after bone marrow transplantation for reducing the graft failure.
Antibodies, Monoclonal↗
[Role of neutrophil and T cell functions in host defense mechanisms of the elderly].
We studied neutrophil functions (phagocytosis, intracellular killing and chemotaxis with or without recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and T cell functions (lymphocyte proliferation and production of GM-CSF in response to phytohemagglutin (PHA)) to clarify host defense mechanisms in the elderly. There was no significant difference in phagocytic activity of neutrophils between the elderly and control young adults. rhGM-CSF enhanced phagocytosis by neutrophils, and a similar degree of enhancement was obtained in both groups. Killing activity of neutrophils evaluated by the new Nitroblue tetrazolium reduction test in the elderly was significantly lower than that in young adults (p < 0.001), however, pretreatment of neutrophils with rhGM-CSF resulted in an increase of killing activity in the elderly, raising their response to a level comparable to that of young adults pretreated with rhGM-CSF. There was no significant difference between the elderly and young adults in chemotaxis of neutrophils. rhGM-CSF alone did not prime chemotaxis, but primed chemotaxis in response to chemoattractant (N-formyl-methionyl-leucyl-phenylalanin) in both individuals. Lymphocyte proliferation and production of GM-CSF in response to PHA in the elderly were significantly lower than those in the young adults (p < 0.001, p < 0.05, respectively). These results indicated that impaired T cell functions may contribute, at least in part, to susceptibility to bacterial infection in the elderly.
Aged↗
[Molecular epidemiology of Chlamydia trachomatis by DNA restriction endonuclease analysis].
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Chlamydia Infections↗
[A case of sarcoidosis presenting with Heerfordt's syndrome, associated with hepatosplenomegaly, pleural effusion, and ascites].
A 30-year-old female developed symptoms consistent with Heerfordt's syndrome (complete type) and was effectively treated with steroid. However, she developed hepatosplenomegaly, bloody pleural effusion, and ascites one year after discontinuation of steroid therapy. The symptoms were considered to be due to sarcoidosis since serum ACE level, and ACE level and OKT4/8 ratio in pleural and peritoneal effusions were significantly elevated, and readministration of steroid normalized these changes. Bloody pleural and peritoneal effusions are very rare complications of sarcoidosis.
Adult↗
Studies on the regulatory domain of Ca2+/calmodulin-dependent protein kinase II by expression of mutated cDNAs in Escherichia coli.
The cDNAs encoding the alpha and beta subunits of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) were ligated into the bacterial expression vector pET and expressed in Escherichia coli. The bacterially expressed alpha and beta subunits exhibited Ca2+/calmodulin-dependent activity and were easily purified to apparent homogeneity from cell extracts. To determine the minimum size required for catalytic activity and the properties of the calmodulin-binding domain, mutated CaM kinase II cDNAs were expressed in E. coli and the enzymatic property of expressed proteins was examined. The replacement of Thr-286 of the alpha subunit with the negatively charged amino acid Asp or that of Arg-283 with the neutral amino acid Gly induced the partially Ca2+ independent activity. The mutant enzymes alpha-I(delta 283-478) and alpha-II(delta 359-478), which truncated the C-terminal region of the alpha subunit, exhibited CaM kinase II activity and the activities of alpha-I(delta 283-478) and alpha-II(delta 359-478) were completely independent of and partially dependent on Ca2+ and calmodulin, respectively. However, the truncated protein alpha(delta 250-478), which was only 33 amino acids shorter than the alpha-I(delta 283-478) protein had no enzymatic activity, indicating that alpha-I(delta 283-478) was close to the minimum size of the active form. The mutant enzyme alpha(delta 291-315), which lacked the calmodulin-binding domain exhibited Ca2+ independent activity. The molecular mass was, however, smaller than that expected from the amino acid sequence. The mutant enzyme alpha(delta 304-315), which lacked the C-terminal half of the calmodulin-binding domain of the alpha subunit, however, exhibited Ca(2+)-independent activity without a reduction in molecular size, indicating that residues 304-315 of the alpha subunit constituted the core calmodulin-binding domain.
Amino Acid Sequence↗
Elevation of nerve growth factor synthesis by constitutive expression of v-src oncogene in cultured rat fibroblasts.
Rat fibroblast 3Y1 cells transformed by Rous sarcoma virus (RSV) or transfected with the v-src gene showed a highly constitutive v-src gene expression. Simultaneously, marked increases in the cellular level of nerve growth factor (NGF) mRNA and NGF content in the culture medium were observed. The levels of NGF mRNA and NGF secreted into the medium were correlated with the expression level of v-src mRNA gene in both transformants and control 3Y1 cells. These results suggest that v-src gene expression is relevant to regulation of NGF synthesis in rat 3Y1 fibroblasts.
Animals↗
Crystallization and characterization of a lectin obtained from a mushroom, Aleuria aurantia.
A fucose-specific lectin with a unique sugar recognizing property was purified from an orange peel mushroom, Aleuria aurantia, by using a specific affinity adsorbent prepared from L-fucose and starch. From 100 g of fruiting bodies, 145 mg of pure lectin was obtained. The lectin was crystallized and the crystals showed hexagonal bipyramid in shape. Distribution of hydrophobic and hydrophilic regions in the molecule of this lectin was predicted from the amino acid sequence deduced from the previously reported nucleotide sequence of the lectin cDNA. Circular dichroism spectra revealed a very low content of alpha-helical and beta-sheet structures and a relatively high content of turns in this lectin. From the spectrum observed in the presence of L-fucose, a hapten sugar of this lectin, certain conformational change was assumed to occur.
Basidiomycota↗
[Studies on uptake of macrolide antibiotics by Chlamydia host cell].
Minimum inhibitory concentration (MIC) of Macrolide antibiotic (ML) against C. trachomatis was found to greatly vary with the cell culture system used for the assay. We then investigated the ability of various cell cultures to uptake MLs in the relation to MIC determination. Penetration of the 14C-labeled MLs, Erythromycin, Jasamycin and Rokitamycin into cells was quantitatively studied by measuring the radioactivity incorporated into McCoy, HeLa229W and HeLa229F cells. It was found that HeLa229W cell showed the lowest MIC for the drugs, followed by HeLa229F cell and then by McCoy cell. Reversely, McCoy cell showed the lowest intracellular concentration of an ML, followed by HeLa229F cell and then by HeLa229W cell. These results indicate that MIC of an ML significantly varies depending on the ability of the test cell to uptake the drug.
Anti-Bacterial Agents↗
[Isolation and identification of Chlamydia spp. from effusion of otitis media].
Three chlamydial strains isolated from patients of otitis media with effusion were studied by comparing reactivity to monoclonal antibody (MAb) and polyclonal antibody (PAb) produced against one clinical isolate (named Mk), which was first isolated by Dr Mukai (Mukai Microbiological Research Laboratory, Yamato-shi, Kanagawa prefecture). Commercially supplied antibody (Microtrak (Syva), Culture-set (Ortho diagnostic system)) was also used. To isolate the Chlamydia spp, the yolk sacs of eggs were immediately inoculated with sample effusions (0.2 to 0.4 ml per sac) as soon as the samples were received. The eggs were observed every day for a period of 12 days thereafter for signs of life or death. One to two blind passages were first done in the eggs and then in HeLa 229 cells. The reactivity was examined by both micro-IF tests, among various strains of Chlamydia (C. trachomatis: L2. C. pneumoniae, C. psittaci: Budgerigar, Izawa, Meningopneumonitis (MP)) and by immunoblot analysis. Chlamydia spp were isolated in two of the twenty-nine sample effusions (6.9%). These isolates were then tested for reactivity to MAb and PAb. It was found that MAb reacted with MP and Mk, but not with Budgerigar, Izawa and C. pneumoniae. The antibody of Culture-set reacted with C. trachomatis C. pneumoniae and C. psittaci. No reactivity was observed in Mk by MicroTrak. Immunoblot analysis revealed that MAb reacted with about 95 KDa protein of Mk, the two clinically isolated Chlamydia spp and MP. By using PAb from rabbits, similar blotting patterns were observed in Mk, the clinical isolates and MP.(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗