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Biomedical subjects

T Kitamura

Publications and source records attributed to T Kitamura.

At least 433 records · Page 24Linked to original sources

[Rearing related stress and depressive severity].

Mothers' difficulties concerning child-rearing were conceptualized as a rearing-related stress (RRS). Eight hundred and seventeen mothers who had six month-olds infants were asked to rate 28 RRS items and 20 items on the depressive severity scale (Zung, 1956). The main results were summarized as follows: (1) Twenty-two items of RRS were analyzed by Hayashi's quantification (type-III) method, and two hypothesized dimensions were extracted. These are named children-related rearing stress (CRRS) and mothers-related rearing stress (MRRS). (2) RRS was related to mothers depressive severity. (3) Linear relationships of "CRRS-MRRS-depression severity" was examined by partial correlation analysis. (4) Primiparae experienced more RRS than multiparae. These results suggested that RRS could be considered as a process, i.e., CRRS influences MRRS and then MRRS influences depressive severity. The RRS model is an accord with the psychological stress model of Lazarus and Folkman (1986). Lastly, possible preventive strategies for mothers' RRS were discussed in the light of RRS model.

Adolescent↗

[A case of pulmonary eosinophilic granuloma with extrapulmonary involvement treated effectively with steroid hormone].

A 21-year-old female was admitted to Tamono City Hospital with chief complaint of dyspnea due to spontaneous pneumothorax. The patient was transferred to our clinic for further examination of abnormal chest X-ray findings together with skin lesions and other symptoms of polyuria, amenorrhea and narrowed visual fields. Histochemical and electron microscopic examination of lung tissue obtained by open lung biopsy led to the diagnosis of eosinophilic granuloma. Furthermore, enhanced computed tomography and magnetic resonance images showed the existence of an intracranial tumor in the supra-hypophyseal region which was interpreted as the cause of hormonal and visual disturbances. Because of dyspnea and enlargement of intracranial tumor, treatment with steroid hormone was commerced followed by tapering of the dose. Chest X-ray findings and skin lesions improved markedly and the intracranial tumor regressed significantly with this steroid therapy, indicating that the intracranial tumor represented involvement with eosinophilic granuloma.

Adult↗

Regulation of hsp70 induction in thermotolerant HeLa cells.

Upon exposure to heat shock, non-thermotolerant (NT) HeLa cells transiently synthesize a large amount of 70-kDa heat-shock protein (hsp70), whereas thermotolerant (TT) cells synthesize a small amount of hsp70. When the hsp70 mRNA of HeLa cells was analyzed, it became apparent that hsp70 mRNA in TT cells did not increase following heat shock, whereas hsp70 mRNA in NT cells did increase dramatically. A further analysis of the activation of the heat-shock transcription factor (HSF) showed that significant activation of HSF was observed immediately after heat shock in both NT and TT cells. However, activated HSF was rapidly repressed in the TT cells, but not in the NT cells. Thus, the decreased induction of hsp70 synthesis observed in the TT HeLa cells may be due to the immediate repression of activated cellular HSF, which probably results in the reduced induction of hsp70 mRNA. The hsp70 content in the TT cells was usually higher than in the NT cells. However, after heat-shock treatment, the hsp70 content of the NT cells increased to nearly the level of the TT cells concomitant with the repression of hsp70 synthesis. The association of activated HSF with hsp70 was observed in both NT and TT cells, and the amount of HSF-hsp70 complex within the cell increased in proportion to the increase in hsp70 in the cells. These findings strongly suggest that the activity of HSF is negatively regulated by the intracellular content of hsp70 in these cells. Furthermore, in vitro experiments on the activation of HSF suggest that HSFs of NT and TT cells may have different properties, or an unknown factor may exist which regulates HSF activation in these cells.

Base Sequence↗

Allocation of mouse cerebellar granule cells derived from embryonic ventricular progenitors--a study using a recombinant retrovirus.

Granule cells of the mammalian cerebellar cortex originate from embryonic progenitors present in the ventricular germinal layer. To investigate the allocation fate of these ventricular progenitors in the mouse, we labeled a few of them on embryonic day 13 with a recombinant retrovirus carrying lacZ which encodes E. coli beta-galactosidase (beta-gal), and the labeled cells in the postnatal cerebellar cortex were detected by beta-gal histochemistry. In the postnatal cerebellar cortex, the virally-labeled beta-gal+ granule cells formed discrete clusters. These clusters were not compactly packed with the beta-gal+ cells, and there was intermingling with beta-gal- granule cells. Neither beta-gal+ Purkinje cells nor glia were found to be included in the clusters. Most of the granule cell clusters were incompatible with the functional areas of the cortex. These results suggest: (1) granule cells derived from individual ventricular progenitors are allocated in clusters and are not extensively dispersed, (2) granule cells descended from one progenitor may mix with their neighbors that are descended from another progenitor, (3) the allocation fate of the ventricular progenitors of granule cells is not restricted to the functional areas of the cerebellar cortex.

Animals↗

Expression and factor-dependent modulation of the interleukin-3 receptor subunits on human hematopoietic cells.

Interleukin-3 (IL-3) regulates growth and differentiation of multipotential as well as lineage-committed progenitor cells. The human IL-3 receptor (IL-3R) consists of the alpha and common beta (beta c) subunits. The alpha subunit (IL-3R alpha) is specific for IL-3 and binds IL-3 with low affinity. In contrast, the beta c subunit does not bind any cytokine by itself, but forms a high-affinity receptor with IL-3R alpha. As the same beta c subunit also forms high-affinity receptors for IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF) with the respective cytokine-specific alpha subunit, the expression of the alpha subunits is responsible for specificity of cytokines. To examine the expression of IL-3R alpha, we have developed a monoclonal antibody (MoAb), N3A. N3A specifically bound to cells expressing IL-3R alpha and immunoprecipitated a 75 Kd glycoprotein, which became 43 Kd on N-glycosidase digestion. N3A and an anti-beta c antibody, CRS1, were used in double color fluorescence-activated cell sorter (FACS) staining with several lineage markers to see the IL-3R expression pattern in peripheral blood (PB), cord blood (CB), and bone marrow (BM) cells. Both IL-3R subunits were expressed on myeloid cell lineages (CD13+, CD14+, CD15Lo, or CD33+). To further study the IL-3R expression on hematopoietic progenitor cells, the CD34+ populations were isolated from both BM and CB cells. Those populations showed positive staining profiles with the N3A MoAb and were weakly stained with the CRS1 MoAb. Furthermore, anti c-kit antibody staining of the CD34+ fraction from CB, but not from BM, showed two intensities and the IL-3R alpha expression seemed to be higher in a fraction of low c-kit expression. Because IL-1, IL-6, G-CSF, stem cell factor (SCF), interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha are known to enhance IL-3-dependent colony formation, we have examined whether this enhancement could be correlated with upregulation of the IL-3R expression. Incubation of CD34+ cells with TNF-alpha for 2 days significantly increased the level of beta c and G-CSF increased the number of cells with high level expression of alpha, while other factors did not affect the IL-3R expression. Thus, different cytokines appear to have different mechanisms for enhancement of IL-3-dependent proliferation.

Antibodies, Monoclonal↗

Ligand-dependent activation of chimeric receptors with the cytoplasmic domain of the interleukin-3 receptor beta subunit (beta IL3).

beta IL3 (formerly known as AIC2A), a beta subunit of the murine interleukin-3 receptor (IL-3R), is not only required for formation of the high affinity receptor but is also important for signal transduction. To examine the function of beta IL3 in signal transduction, we constructed several chimeric receptors consisting of the intracellular portion of beta IL3 and the extracellular portion of other members of the cytokine receptor superfamily, i.e. the human interleukin-2 receptor beta chain (hIL-2R beta), the human interleukin-4 receptor (hIL-4R), and the murine erythropoietin receptor (mEpoR). These chimeric receptors and normal cytokine receptors were expressed in an IL-3-dependent murine pro-B cell line, Ba/F3, and an IL-2-dependent murine T cell line, CTLL2. Regardless of the origin of the extracellular domain, these chimeric receptors were functional in Ba/F3 cells; they stimulated proliferation and induced tyrosine phosphorylation in response to the cytokine corresponding to the extracellular domain. However, the response of transfectants expressing chimeric receptors was similar to, but not identical with, the response of Ba/F3 cells to mIL-3. We present evidence that the IL-4R and EpoR probably have an additional component which is involved in signal transduction.

Animals↗

Risk factors for hepatocellular carcinoma among patients with chronic liver disease.

BACKGROUND AND METHODS: To detect potentially curable cases of hepatocellular carcinoma, outpatients with chronic hepatitis or compensated liver cirrhosis who were seen at the Center for Adult Diseases (Osaka, Japan) were examined periodically by means of ultrasonography and measurement of serum alpha-fetoprotein. Risk factors for hepatocellular carcinoma were identified with a Cox proportional-hazards model. RESULTS: A total of 917 patients, 40 to 69 years old, were registered from May 1987 to March 1991. By the end of September 1991, liver cancer had developed in 54. The three-year cumulative risk of liver cancer was 12.5 percent for 240 patients with liver cirrhosis at enrollment and 3.8 percent for 677 patients with chronic hepatitis. Cox regression analysis showed that the risk of liver cancer was increased almost sevenfold in patients with hepatitis B surface antigen (rate ratio, 6.92; 95 percent confidence interval, 2.92 to 16.39) and fourfold in patients with hepatitis C antibody (rate ratio, 4.09; 95 percent confidence interval, 1.30 to 12.85). A high alpha-fetoprotein value at enrollment was also a risk marker for liver cancer. CONCLUSIONS: Patients with hepatitis C virus infection have a greatly increased risk of liver cancer. Further studies are required to clarify the roles of other risk factors, including drinking and smoking habits.

Adult↗

The structure and dynamics of the granulocyte macrophage colony-stimulating factor receptor defined by the ternary complex model.

Myeloid cell lines and primary leukemic myeloblasts express two classes of granulocyte macrophage colony-stimulating factor (GM-CSF) binding sites of high (Kd 20-50 pM) and low affinity (Kd 5-10 nM). High affinity binding is caused by the association of two chains, p80 alpha and p130 beta, whereas p80 alpha alone confers low affinity binding only. Furthermore interleukin-3 (IL-3) competes for the binding of GM-CSF to its high affinity receptor (for review see Nicola, N. A., and Metcalf, D. (1991) Cell 67, 1-4). In the present study, we took advantage of the perturbation of GM-CSF binding equilibrium by IL-3 to take a quantitative approach to analysis of the structure and dynamics of the GM-CSF receptor complex. First, cross-linking studies were performed at two concentrations of radioligand. At 200 pM, a concentration sufficient for near saturation of the high affinity binding site R1, the association between p80 alpha and p130 beta is stoichiometric, and the addition of IL-3 prevents the binding to both chains. At 5 nM, a concentration sufficient for half-occupancy of the low affinity binding site R2, IL-3 prevents cross-linking to the beta chain only. Second, GM-CSF saturation curves were analyzed both at equilibrium and under conditions of perturbation of the equilibrium by IL-3. In the presence of IL-3, the interaction of GM-CSF with its receptor is converted from high to low affinity binding. Computer modeling of binding data with a ternary complex model involving GM-CSF, p80 alpha, and p130 beta indicates that the model fits the data with accuracy and suggests that ligand binding stabilizes the interaction between p80 alpha and p130 beta by 3 orders of magnitude. Third, membrane solubilization dissociates p80 alpha and p130 beta whereas on ligand-stabilized preformed complexes, solubilization did not dissociate the two chains. Finally, upon addition of GM-CSF, there is an increase with time in the proportion of ligand bound to the high affinity receptor, at the expense of that bound to low affinity receptor, suggesting that stabilization of the ternary complex is a time-dependent process.

Binding Sites↗

Origin of JC polyomavirus variants associated with progressive multifocal leukoencephalopathy.

JC polyomavirus (JCV) DNAs from the urine of nonimmunocompromised individuals (designated archetypal isolates) regularly contain a regulatory sequence that may have generated various regulatory sequences of JCV isolates derived from the brain of patients with progressive multifocal leukoencephalopathy (PML). In this report, we constructed a phylogenetic tree for 14 isolates (7 archetypes and 7 PML types) from DNA sequence data on the VP1 (major capsid protein) gene. According to the phylogenetic tree, the 14 isolates diverged into types A and B, each of which contained archetypal and PML-type isolates. Each type further diverged into several groups containing archetypal and PML-type isolates. We conclude that PML-type isolates are polyphyletic in their origin and do not constitute a unique lineage. This conclusion suggests that PML-type JCV isolates are generated from archetypal strains during persistence in the hosts. Furthermore, the present phylogenetic analysis indicates that an ancestral JCV carried the archetypal regulatory sequence and that this structure has been conserved in the course of JCV evolution.

Base Sequence↗

Reconstitution of the functional receptors for murine and human interleukin 5.

The murine interleukin 5 receptor (mIL-5R) is composed of two distinct subunits, alpha and beta. The alpha subunit (mIL-5R alpha) specifically binds IL-5 with low affinity. The beta subunit (mIL-5R beta) does not bind IL-5 by itself, but forms the high-affinity receptor with mIL-5R alpha. mIL-5R beta has been revealed to be the mIL-3R-like protein, AIC2B which is shared with receptors for IL-3 and granulocyte/macrophage colony-stimulating factor. We demonstrated here the reconstitution of the functional receptors for murine and human IL-5 on the mouse IL-2-dependent cell line, CTLL-2. CTLL-2 was transfected with the cDNAs for mIL-5R alpha and/or AIC2B. Only CTLL-2 transfectant expressing both mIL-5R alpha and AIC2B expressed the high-affinity receptor and proliferated in response to murine IL-5. Then CTLL-2 was transfected with the cDNAs for hIL-5R alpha and/or KH97 (beta c), the human homologue of AIC2B. Though beta c did not contribute much to binding affinity of hIL-5R, only CTLL-2 transfectant expressing both hIL-5R alpha and beta c proliferated in response to human IL-5. These results showed that the beta subunit is indispensable in IL-5 signal transduction. We further investigated the function of IL-5-specific alpha subunit in transmitting IL-5 signals. Mutant mIL-5R alpha, which lacks its whole cytoplasmic domain, was transfected into mouse IL-3-dependent cell line, FDC-P1 expressing AIC2B intrinsically. The resulting transfectant did not respond to IL-5, though the transfectant expressed the high-affinity IL-5R, indicating that the cytoplasmic portion of the alpha subunit also has some important role in IL-5-mediated signal transduction.

Animals↗

Reduced diameter and conduction velocity of myelinated fibers in the sciatic nerve of a neurofilament-deficient mutant quail.

The diameter and conduction velocity of myelinated fibers in the sciatic nerve isolated from neurofilament-deficient quail ('quiver') were compared with those from wild quail. The diameter was significantly reduced in quiver, however, the frequency distribution formed three peaks as did that observed in the wild. The fibers were categorized in three groups possessing different conduction velocity. The conduction velocity appeared to be proportional to the diameter in each strain, and was significantly reduced in quiver. These results indicate that, although the diameter and conduction velocity are reduced in quiver fiber, the correlation between the parameters observed in wild quail is still preserved.

Animals↗

Cells of origin of the spinoparabrachial fibers in the rat: a study with fast blue and WGA-HRP.

The internal lateral (IL) subnucleus of the parabrachial nucleus (PB), which is one of the seven lateral subnuclei of the PB, receives information from the spinal cord. The IL subnucleus perhaps relays nociceptive signals to the intralaminar nuclei of the thalamus, apparently being implicated in the motivational-affective component of pain reaction. However, cells of origin of spinal fibers to the IL subnucleus have not been investigated sufficiently. We intended to clarify these cells by injection of fast blue or wheat germ agglutinin-conjugated horseradish peroxidase into the IL subnucleus and/or other lateral subnuclei in the rat. When the tracer was injected into the IL subnucleus, many cells were labeled bilaterally in laminae I, V, and VII, and in the dorsolateral and dorsomedial parts of the lateral funiculus throughout the entire length of the spinal cord. A small number of labeled cells appeared ipsilaterally in laminae II-IV and VI in the upper cervical segments and contralaterally in laminae VIII and X throughout the spinal cord. Labeled cells in lamina I were more numerous ipsilaterally than contralaterally in the first two cervical segments but were more numerous contralaterally than ipsilaterally in the remaining spinal segments. Labeled cells were seen with a contralateral predominance in lamina VII, but with an ipsilateral predominance in lamina V and in the dorsolateral and dorsomedial parts of the lateral funiculus. With tracer injected into the lateral subnuclei of the PB, excluding the IL subnucleus, labeled cells were found primarily in lamina I throughout the entire length of the spinal cord. These results show that cells giving rise to spinoparabrachial fibers were more numerous and more widely distributed than previously reported.

Afferent Pathways↗

GM-CSF triggers a rapid, glucose dependent extracellular acidification by TF-1 cells: evidence for sodium/proton antiporter and PKC mediated activation of acid production.

The extracellular acidification rate of the human bone marrow cell line, TF-1, increases rapidly in response to a bolus of recombinant granulocyte-macrophage colony stimulating factor (GM-CSF). Extracellular acidification rates were measured using a silicon microphysiometer. This instrument contains micro-flow chambers equipped with potentiometric sensors to monitor pH. The cells are immobilized in a fibrin clot sandwiched between two porous polycarbonate membranes. The membranes are part of a disposable plastic "cell capsule" that fits into the microphysiometer flow chamber. The GM-CSF activated acidification burst is dose dependent and can be neutralized by pretreating the cytokine with anti-GM-CSF antibody. The acidification burst can be resolved kinetically into at least two components. A rapid component of the burst is due to activation of the sodium/proton antiporter as evidenced by its elimination in sodium-free medium and in the presence of amiloride. A slower component of the GM-CSF response is a consequence of increased glycolytic metabolism as demonstrated by its dependence on D-glucose as a medium nutrient. Okadaic acid (a phospho-serine/threonine phosphatase inhibitor), phorbol 12-myristate 13-acetate (PMA, a protein kinase C (PKC) activator), and ionomycin (a calcium ionophore) all produce metabolic bursts in TF-1 cells similar to the GM-CSF response. Pretreatment of TF-1 cells with PMA for 18 h resulted in loss of the GM-CSF acidification response. Although this treatment is reported to destroy protein kinase activity, we demonstrate here that it also down-regulates expression of high-affinity GM-CSF receptors on the surface of TF-1 cells. In addition, GM-CSF driven TF-1 cell proliferation was decreased after the 18 h PMA treatment. Short-term treatment with PMA (1-2 h) again resulted in loss of the GM-CSF acidification response, but without a decrease in expression of high-affinity GM-CSF receptors. Evidence for involvement of PKC in GM-CSF signal transduction was obtained using calphostin C, a specific inhibitor of PKC, which inhibited the GM-CSF metabolic burst at a subtoxic concentration. Genistein and herbimycin A, tyrosine kinase inhibitors, both inhibited the GM-CSF response of TF-1 cells, but only at levels high enough to also inhibit stimulation by PMA. These results indicate that GM-CSF activated extracellular acidification of TF-1 cells is caused by increases in sodium/proton antiporter activity and glycolysis, through protein kinase signalling pathways which can be both activated and down-regulated by PMA.

Benzoquinones↗

Psychological and social correlates of the onset of affective disorders among pregnant women.

One hundred and twenty women recruited from attenders at the antenatal clinic of the Obstetrics Department of a general hospital were asked to complete ad hoc questionnaires during pregnancy; they were then interviewed by psychiatrists using a structured diagnostic interview, the Schedule for Affective Disorders and Schizophrenia (SADS). Nineteen (16%) women were identified as having an onset of an affective disorder during the period of pregnancy, mainly (68%) during the first trimester. As compared with the women without any such onset (controls), the women with pregnancy-related affective disorder (PRAD) were characterized by (1) it being their first pregnancy or first delivery with past termination of pregnancy, (2) early loss of either parent by death, (3) high Eysenck Personality Questionnaire (EPQ) Neuroticism (N) and Psychoticism (P) scores, (4) living in a flat with either a plan to stay there after the forthcoming childbirth or an expectation that their accommodation would be crowded, and (5) negative response to the news of the pregnancy by the husband with low intimacy. The effects of these factors were additive since the probability of developing a PRAD episode was highly correlated with the number of factors reported.

Adolescent↗