Biomedical subjects
Publications and source records attributed to T Murase.
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The plasma concentration of arginine vasopressin (AVP) is closely regulated by plasma osmolality. In this study, we used intronic in-situ hybridization to investigate the transcriptional activity of the AVP gene in the supraoptic nucleus (SON) and paraventricular nucleus (PVN) after intraperitoneal (i.p.) injection of hypertonic saline inducing small changes in plasma osmolality in rats. After ip injection of 900 mOsm/kg saline (2% BW), plasma [Na] reached the highest level at 10 min (900 mOsm/kg, 146+/-1 mEq/l; 290 mOsm/kg, 139+/-1 mEq/l, P<0.01) and maintained that level until 30 min. The expression of AVP heteronuclear (hn) RNA in the SON and PVN increased significantly as early as 10 min and peaked 30 min after ip injection of the hypertonic saline. AVP hnRNA expression showed a significant increase even after a 2 mEq/L elevation in plasma [Na] concentration, and increased dose-dependently in response to the concentration of saline injection (290-900 mOsm/kg). These results demonstrate that, similar to AVP secretion, AVP gene transcriptional activity in the SON and PVN is closely regulated by plasma osmolality.
Overexpression and mutation of p53 in 46 primary endometrial carcinomas were determined comparatively with the status for estrogen receptor (ER) and progesterone receptor (PR). Immunohistochemically p53 overexpression was found in 9 of 46 cases (20%), while eight kinds of mutations in that gene were detected in 7 of 46 endometrial carcinomas (15%), using polymerase chain reaction single-strand conformation polymorphism and subsequently direct sequencing method. Six of nine cases with p53 overexpression showed p53 mutation. All eight mutations showed single substitutions, and three cases in exon 4, one in exon 5, two in exon 6, and two in exon 7 were found. The cases with the p53 overexpression were significantly inversely related to that of ER or PR staining. Most endometrial carcinoma with p53 overexpression and/or mutation had a relatively poor prognosis and showed no reactivities of ER or PR.
CD5, first recognized on subsets of lymphocytes, also is detected in thymic carcinoma but not in thymoma or other malignant tumors. We studied CD5 expression in 73 cases of malignant tumors of various organs, 22 cases of thymoma, and 7 cases of thymic carcinoma by immunohistochemistry using the new monoclonal anti-CD5 antibody, NCL-CD5-4C7, with a pressure cooker antigen retrieval method. All cases of thymic carcinoma showed positive staining for CD5, predominantly on the cell membrane. Two of 4 cases of atypical thymoma also showed focal positivity, whereas the other types of thymoma were negative. CD5 was detected in cases of malignant tumors other than squamous cell carcinoma and in the normal epithelium of their counterparts. Squamous cell carcinomas of various organs were negative for CD5. Malignant mesothelioma showed peculiar intracytoplasmic staining in contrast to the other tumors. The NCL-CD5-4C7 positivity in thymic epithelial tumors may support the hypothesis suggesting progression of atypical thymoma to thymic carcinoma. NCL-CD5-4C7 may be useful in the differential diagnosis of mediastinal tumors, especially between thymic carcinoma and metastatic squamous cell carcinoma of various primary sites, and for distinguishing malignant mesothelioma from adenocarcinoma of the lung by the different staining pattern.
Identification of the SYT-SSX fusion transcript can be a useful diagnostic aid for synovial sarcoma. However, there is no report on the detection of the specific transcript in archival cytologic specimens. We performed a rapid reverse transcription-polymerase chain reaction assay to detect the fusion transcript using such specimens as RNA sources and compared its usefulness with the assay using formalin-fixed paraffin-embedded histologic specimens. Eleven cytologic and 12 histologic specimens stored for 0.1 to 20 years were selected from 9 patients. The SYT-SSX transcript was identified in 10 (91%) of 11 of cytologic and 9 (75%) of 12 histologic cases. One of the transcript-positive cytologic cases was obtained by fine-needle aspiration. SYT-SSX1 fusion was found in 4 patients and SYT-SSX2 in 5. Cytologic specimens will be an excellent RNA source for the reverse transcription-polymerase chain reaction diagnosis of synovial sarcoma with a sensitivity of more than 90%, comparable to that in frozen materials. Histologic materials also may be useful when cytologic or frozen materials are not available.
Shiga toxin-producing Escherichia coil (STEC) O157 were isolated from processed salmon roe which had been a suspected food item in sporadic infections which occurred in Japan in 1998. A total of 45 samples of the processed salmon roe were pre-enriched in trypticase soy broth (TSB) at 36 degrees C for 6 h and novobiocin-supplemented modified EC broth (mEC-NB) at 42 degrees C for 18 h. After the pre-enrichments, the cultures were examined for possible occurrence of STEC O157, using an immunomagnetic separation (IMS) method. From the examination, a total of 84 strains of STEC O157:H7 that were positive for both stx 1 and stx 2 genes were isolated. By applying the most-probable-number technique, it was estimated that the number of STEC O157 was in the range of 0.73-1.5 per 10 g of the processed salmon roe. Subsequent analysis of the isolates by a pulsed-field gel electrophoresis (PFGE) revealed a pattern commonly seen in 82 isolates and another pattern in two isolates. Clinical isolates from 7 patients also showed an identical pattern to those of the 82 isolates and one isolate from a patient showed the other pattern identical to those of the two isolates. The isolates were found to belong to the phage type 14.
We report a 67-year-old male with Waldenstrom's macroglobulinemia who developed Campylobacter fetus subspesis fetus (C. fetus) bacteremia and thrombophlebitis. The patient developed a fever and pain in his left lower limb, and could not walk because of the pain. Radioisotopic venography showed thrombophlebitis in his left lower limb. His blood culture grew C. fetus. After starting intravenous PAPM/BP, his symptoms resolved promptly. In contrast to Campylobacter jejuni which is a common cause of infectious diarrhea, C. fetus infection has distinct clinical features showing systemic illness such as bacteremia and thrombophlebitis mainly occurring in immunocompromized patients. This organism should be considered as one of the possible pathogenes in the infectious complications of the immunocompromized patients.
Streptococcus pyogenes isolates obtained in 1981 to 1997 from patients and healthy subjects were characterized by pulsed-field gel electrophoresis (PFGE) patterns, biotyping, and the presence of spe genes encoding streptococcal pyrogenic exotoxins. Changes in the profiles were shown in the serotype M1/T1 isolates from pharyngitis over this period, but not in serotype M3/T3 isolates. The characteristics of isolates from patients with toxic shock-like syndrome (TSLS) were comparable to those of the other isolates, including those from healthy subjects. This finding suggests that further phenotypic and molecular characterization, such as investigating the genomic difference represented by the pathogenicity island, of isolates with apparently the same profiles would be necessary to determine the etiology of diseases caused by S. pyogenes, including TSLS.
Gallates (gallic acid esters) belong to the class of phenolic compounds, which are abundant in red wine. In this study, we show that gallates can inhibit cytokine-induced activation of nuclear factor kappaB (NF-kappaB) and thereby reduce expression of endothelial-leukocyte adhesion molecules in cultured human umbilical vein endothelial cells (HUVECs). Pretreatment of HUVECs with ethyl gallate (3 to 10 micromol/L) significantly suppressed interleukin-1alpha (IL-1alpha)- or tumor necrosis factor-alpha (TNF-alpha)- induced mRNA and cell-surface expression of vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and E-selectin, which was associated with reduced adhesion of leukocytes to HUVECs. Gel shift assays with the NF-kappaB consensus sequence showed the decreased densities of the shifted bands in gallate-treated HUVECs. Furthermore, gallate pretreatment inhibited cytokine-induced transcription of a fusion gene, which consisted of 4 repeats of the NF-kappaB consensus sequence and the luciferase reporter gene. Immunoblot analysis of nuclear extracts and whole-cell lysates demonstrated the decreased amounts of NF-kappaB p65 in nuclei but equal amounts of inhibitor-kappaBalpha (I-kappaBalpha) in whole-cell lysates of ethyl gallate-treated HUVECs. Incubation of the nuclear extracts from cytokine-activated HUVECs with ethyl gallate did not affect the NF-kappaB shifted bands induced by cytokines in gel shift assays. Taken together, these data demonstrate that ethyl gallate can inhibit cytokine-induced nuclear translocation of NF-kappaB p65 by way of a mechanism independent of I-kappaBalpha degradation and thereby suppress expression of VCAM-1, ICAM-1, and E-selectin, which was associated with reduced adhesion of leukocytes. These results in vitro demonstrate that gallates can exhibit anti-inflammatory properties by blocking activation of NF-kappaB and suggest that these natural compounds, abundant in red wine, may play important roles in the prevention of atherosclerosis and inflammatory responses in vivo.
The aim of this study is to identify insulin-dependent diabetes mellitus (IDDM)-susceptible HLA antigens in IDDM patients who do not have established risk allele, HLA-DQA1*0301, and analyze relationship of these HLA antigens and the degree of beta-cell destruction. In 139 Japanese IDDM patients and 158 normal controls, HLA-A, -C, -B, -DR and -DQ antigens were typed. Serum C-peptide immunoreactivity response (deltaCPR) to a 100-g oral glucose load < or = 0.033 nmol/l was regarded as complete beta-cell destruction. All 14 patients without HLA-DQA1*0301 had HLA-A24, whereas only 35 of 58 (60.3%) normal controls without HLA-DQA1*0301 and only 72 of 125 (57.6%) IDDM patients with HLA-DQA1*0301 had this antigen (Pc = 0.0256 and Pc = 0.0080, respectively). DeltaCPR in IDDM patients with both HLA-DQA1*0301 and HLA-A24 (0.097 +/- 0.163 nmol/L, mean +/- SD, n = 65) were lower than in IDDM patients with HLA-DQA1*0301 only (0.219 +/- 0.237 nmol/L, n = 45, P < 0.0001) and in IDDM patients with HLA-A24 only (0.187 +/- 0.198 nmol/L, n = 14, P = 0.0395). These results indicate that both HLA-DQA1*0301 and HLA-A24 contribute susceptibility to IDDM independently and accelerate beta-cell destruction in an additive manner.
Recent results indicate that renal escape from vasopressin-induced antidiuresis is accompanied by a marked downregulation of whole kidney aquaporin-2 (AQP-2) protein and mRNA expression. However, in those studies, the escaped animals were also markedly hypo-osmolar compared to controls as a result of water loading during antidiuresis. The present studies evaluated whether systemic or local osmolality contributes to the downregulation of AQP-2 expression in this model. In the first study, two groups of 1-deamino-[8-D-arginine]-vasopressin (dDAVP)-infused rats were water-loaded; after establishment of escape, one group was then water-restricted for 4 d to reverse the escape, whereas the other group continued daily water loading. Whole kidney AQP-2 protein was measured by Western blotting, and inner medulla AQP-2 mRNA was determined by Northern blotting. Results were compared to dDAVP-infused rats fed solid chow. After 4 d of water restriction, urine volume decreased to the same level as in the rats on solid chow; however, plasma sodium concentrations and plasma osmolality remained low. Despite maintenance of significant hypo-osmolality, rats in which escape was subsequently reversed by water restriction reestablished high dDAVP-stimulated kidney levels of AQP-2 after 4 d of water restriction. In the second study, AQP-2 expression was evaluated in different regions of kidneys from water-loaded rats undergoing escape from antidiuresis. Despite markedly different interstitial osmolalities, significant downregulation of AQP-2 expression compared to dDAVP-infused control rats was seen in the inner medulla, outer medulla, and cortex. Thus, neither systemic nor interstitial osmolality appears to appreciably be correlated with downregulation of kidney AQP-2 expression during escape from antidiuresis. These results therefore suggest that additional vasopressin- and osmolality-independent factors, likely related to the effects of extracellular fluid volume expansion, also regulate kidney AQP-2 expression in rats.
The present study investigated the validity, reliability, objectivity, and practicability of methods with skinfold caliper (SF), ultrasound (US), bioelectrical impedance (BI), and sulfur hexafluoride (SHD) using identical subjects, and evaluated comprehensive usefulness by comparing selected methods or equations. In examining validity, underwater weighing (UW) was employed to obtain the criterion of validity. The subjects were healthy Japanese, 16 males and 15 females, aged 18 to 32 years. The prediction equation developed by Nakadomo et al. (1990b) was considered to be suitable for BI. With respect to the validity, BI and SHD assessing total body when estimating body composition would be more valid than those assessing partial subcutaneous skinfold thickness. A comprehensive evaluation through an examination with respect to the validity, reliability, objectivity, and practicability suggests that the BI could be the best method to assess human body composition in vivo.
Malignant histiocytosis-like B-cell lymphoma (MH-like BCL) is characterized clinically by hemophagocytic syndrome (HPS), bone marrow involvement at presentation, and an aggressive clinical course. Yet, it remains an ill-defined disease entity. We recently described five cases of MH-like BCL and suggested that these may be regarded as a peculiar variant of intravascular lymphomatosis (IVL), based on clinical and pathological observation. Interestingly, this type of lymphoma has been reported exclusively among Asians, with few reports from Western countries, and therefore we propose that this variant be noted as an Asian variant of IVL(AIVL). To evaluate the incidence of this variant, we performed a literature search for English or Japanese studies dealing with MH-like BCL or B-cell IVL associated with HPS (IVL with HPS). We found 18 reports of MH-like BCL and 16 reports of IVL with HPS and re-evaluated the clinicopathologic aspects of each study with respect to AIVL. Although several differences were noted, such as gender, presence of respiratory changes, bone marrow invasion and disseminated intravascular coagulopathy, there were also overlapping profiles and they appeared to be consistent with the AIVL single disease entity. Of particular note was the absence of peripheral lymphadenopathy and tumor formation which posed diagnostic problems in the initial phase of the presentations. Thus, based on analysis of the clinical findings of these 34 cases, the diagnostic criteria for AIVL consists primarily of cytopenia (anemia and/or thrombocytopenia), hepatosplenomegaly, absence of peripheral lymphadenopathy and tumor formation, and erythrocyte-hemophagocytosis along with intravascular proliferation of the neoplastic B cells. The pathologic findings of the 19 autopsied cases which fulfilled the criteria are also summarized, and they reveal morphologic, immunologic, and genotypic features of typical IVL.
In order to obtain the realistic background information on clinical features, and the present status of treatment and outcome in elderly patients with acute lymphoblastic leukemia (ALL), we carried out random survey of patients with ALL aged 60 or over who had been admitted to 13 general hospitals in the Nagoya region from January 1990 through December 1995. Among the 20 cases collected, ages ranged from 60 to 88 (median age 68), and the male to female ratio was 11:9. Nineteen cases were L2 subtype in FAB classification. Among 17 patients, 13 had B cell series surface phenotypes (76%), 2 had T cell series (12%), one had stem cell type (6%) and one had an undetermined phenotype (6%). Ph chromosomes were detected in 4 cases among 15 analyzed (27%), whereas 5 were found to have no chromosomal abnormality. Half of the patients had some concurrent disease at diagnosis, including two with treatment-limiting complications. Common induction regimens were the combination of adriamycin (ADM) + vincristine (VCR) + cyclophosphamide (CPM) + mitoxdn trone + L-asparaginase [4 patients]. ADM + VCR + PSL [4 patients]. VCR + PSL [4 patients] and others [8 patients]. The overall remission rate was 55.0% (11/20) without any significant difference according to age. The median survival time (MST) for all cases was 205 days. (1-year survival rate:17.9%, 2-year survival rate:10.8%). There was no significant difference in survival times among patients with the Ph chromosome, those with other chromosomal abnormalities and those without them. All the patients aged 75 or over were treated with attenuated induction therapy, and they had a shorter survival than those aged less than 75, but with no statistical significance [MST:121 days versus 276 days, p = 0.307 (generalized Wilcoxon test)].
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A case of mucinous tumor of the gallbladder with a separate nodule of anaplastic carcinoma is reported. The patient was an 83-year-old Japanese man who underwent cholecystectomy under the preoperative diagnosis of a mucus-producing gallbladder tumor. A mucinous tumor was found in the neck and distal body of the gallbladder, associated with a separate nodule in the fundus. The latter nodule was initially diagnosed as a benign xanthogranulomatous lesion. However, the immunohistochemical study revealed that the atypical cells in the superficial part of the nodule were positive for cytokeratin and epithelial membrane antigen, confirming the diagnosis of anaplastic carcinoma. Although the occurrence of mural nodules in mucinous cystic tumors of the ovary and pancreas is well reported, to our knowledge, this is the first report on the occurrence of a mucinous tumor with a nodule of anaplastic carcinoma in the gallbladder.
We report a patient with Philadelphia chromosome (Ph1)-positive chronic myelogenous leukemia (CML) which transformed into blast crisis bearing the immunophenotypic features similar to those of the myeloid/natural killer (NK) cell precursor leukemia we proposed previously. Using a CD45 blast gating method, the myeloperoxidase-negative blasts were positive for CD7, CD13, CD33, CD34, CD56, and HLA-DR, but no other lymphoid antigens. Southern blot analysis showed germ line T cell receptor beta and delta genes and immunoglobulin heavy and light chain genes. Although NK cell blastic transformation with Ph1 positive CML has been reported in a single patient, this is, to our knowledge, the first report of CML blast crisis of myeloid/NK cell precursor origin.