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Biomedical subjects

S Haraguchi

Publications and source records attributed to S Haraguchi.

At least 55 records · Page 3Linked to original sources

[Aneurysmal bone cyst of the left 5th rib].

A case of aneurysmal bone cyst of the left 5th rib is reported. A 57-year-old male complained of palpable lump and pain on his back. Chest X ray and CT showed expansive tumor arid pathological fracture of the left 5th rib. The tumor and the adjacent segments of the 4th and 5th ribs were resected en bloc. The pathological examination showed multiple blood filled spaces and fibrous trabeculae, confirming the diagnosis of aneurysmal bone cyst. He has been followed up for 3 years and 6 months with no signs of recurrence.

Bone Cysts, Aneurysmal↗

Transcriptional down-regulation of tumor necrosis factor-alpha gene expression by a synthetic peptide homologous to retroviral envelope protein.

We have previously shown that a synthetic peptide (CKS-17) homologous to retroviral envelope protein suppresses the accumulation of superantigen staphylococcal enterotoxin-induced TNF-alpha mRNA in human PBMC and in highly purified human monocytes. The present study was designed to examine the underlying mechanism(s) by which CKS-17 down-regulates the TNF-alpha mRNA expression using a human acute monocytic leukemia cell line THP-1 stimulated with the superantigen staphylococcal enterotoxin E. A cyclooxygenase inhibitor indomethacin does not reverse the inhibition of TNF-alpha mRNA expression by CKS-17, suggesting that prostaglandins are not responsible for the suppressive action of CKS-17. The inhibitory effect of CKS-17 is, however, significantly blocked by a protein synthesis inhibitor cycloheximide, indicating that CKS-17 requires de novo protein synthesis to induce the suppressive activity. The mRNA stability assays using actinomycin D show that CKS-17 does not decrease the TNF-alpha mRNA stability. Nuclear run-on transcription assays further reveal that CKS-17 suppresses the TNF-alpha mRNA transcription rate. Taken together, these results suggest that the synthetic retroviral peptide CKS-17 down-regulates TNF-alpha mRNA expression through inhibition of transcriptional activation of the TNF-alpha gene, which requires de novo synthesis of a transcriptional repressor protein(s).

Amino Acid Sequence↗

Prolactin acts on the extreme 5' portion of MMTV LTR involving a mammary cell-specific enhancer.

We have previously shown that a human mammotropic polypeptide hormone, prolactin (PRL) can act synergistically with steroid hormones to regulate gene expression directed by the long terminal repeat of mouse mammary tumor virus (MMTV LTR) in a human ductal carcinoma cell line T47D cells using a chloramphenicol acetyltransferase reporter gene system and gene transfection methods. In the present study, using various recombinant plasmids we analyzed functional elements in the MMTV LTR that is essential for the PRL responses. We show that the PRL-responsive elements are located in the extreme 5' end of the MMTV LTR, a region previously described by others to be a mammary cell-specific enhancer.

Breast↗

Histogenesis of abnormal elastic fibers in blebs and bullae of patients with spontaneous pneumothorax: ultrastructural and immunohistochemical studies.

Elastic fibers in 15 blebs and 17 bullae with spontaneous pneumothorax were studied by means of electron microscopy and light and electron microscopic immunohistochemistry for elastin and alpha 1-antitrypsin. Blebs were formed in association with focal organized alveoli, and bullae were formed in association with pulmonary emphysema. Both blebs and bullae had abnormal elastic fibers. Ultrastructurally, abnormal elastic fibers of blebs and bullae consisted of accumulated thick and fine fibers. Accumulated thick elastic fibers showed vacuolar changes and electron-dense granular deposits, and they were associated with spiraling collagen fibrils. These thick elastic fibers reacted evenly with antielastin antibody and also reacted with anti-alpha 1-antitrypsin antibody. They are thought to be degraded elastic fibers. Accumulated fine elastic fibers consisted of bundles of microfibrils and granular amorphous components, and they reacted with anti-elastin and anti-alpha 1-antitrypsin antibody. These fine elastic fibers are thought to be not only newly formed in the process of organization but also degraded. It is suggested that elastic fibers of blebs and bullae are degraded due to an imbalance between elastase and alpha 1-antitrypsin.

Adult↗

Purification and characterization of a novel growth factor (FF-GF) synthesized by a rat hepatoma cell line, FF101.

A rat hepatoma cell line, FF101, established in serum-free, protein-free medium, synthesizes a growth factor(FF-GF). FF-GF was purified by gel filtration chromatography, cation-exchange chromatography and isoelectric focusing. Purified FF-GF was revealed as a single band on SDS-gel electrophoresis and its molecular weight was estimated to be 70 KDa. FF-GF stimulated DNA synthesis of various cells from different origins. The growth-promoting activities of FF-GF were abolished by treating with protease, dithiothreitol, acid and heating, whereas its activity was not inhibited by antibodies against acidic and basic fibroblast growth factors. These results indicate that FF-GF is a novel growth factor.

Animals↗

Human prolactin regulates transfected MMTV LTR-directed gene expression in a human breast-carcinoma cell line through synergistic interaction with steroid hormones.

Prolactin plays a key role in the regulation and growth of mammary cells, and influences tumor promotion. We have shown that chronic energy restriction intake depresses prolactin levels, inhibits production of MMTV proviral DNA and proto-oncogene expression in mammary glands and prevents development of mammary tumors. Since the expression and proto-oncogene activation of MMTV are regulated by promoter/enhancer elements within its long terminal repeat (LTR), in the present study we used a chloramphenicol acetyl transferase (CAT) reporter gene system and gene transfection methods to study the effect of prolactin on MMTV LTR using a human ductal carcinoma cell line T47D stably or transiently transfected with a plasmid consisting of the LTR upstream of CAT gene. Human prolactin or dexamethasone induced, respectively, a 2-fold or 6-fold increase in CAT activity compared with background CAT activity in the absence of hormones. However, the combination of human prolactin and dexamethasone strongly enhanced (20-fold) induction of the LTR compared with the control. Human prolactin also showed a synergistic effect with progesterone on LTR induction. Both LTR and CAT genes needed to be linked for induction of CAT activity by prolactin and dexamethasone. Our results indicate that human prolactin can act synergistically with steroid hormones to regulate MMTV LTR-directed gene expression in transfected T47D cells.

Breast Neoplasms↗

A synthetic peptide homologous to retroviral envelope protein down-regulates TNF-alpha and IFN-gamma mRNA expression.

We investigated the influence of CKS-17, a synthetic heptadecapeptide that corresponds to a highly conserved domain of the immunosuppressive retroviral envelope protein p15E, on staphylococcal enterotoxin B (SEB)-induced TNF-alpha gene expression in human peripheral blood mononuclear cells and highly purified human monocyte preparations, as well as the production of TNF-alpha protein, using human peripheral blood mononuclear cells. RNA hybridization studies show that CKS-17 inhibits SEB-induced TNF-alpha mRNA accumulation in human peripheral blood mononuclear cells and human monocytes. CKS-17 is also shown to be highly suppressive for SEB-induced production of TNF-alpha proteins. Similarly, CKS-17 inhibits expression of SEB-induced IFN-gamma mRNA in human peripheral blood mononuclear cells. These results suggest that CKS-17 down-regulates both TNF-alpha and IFN-gamma production at mRNA level.

Amino Acid Sequence↗

Suppression of human interferon-gamma production by a 17 amino acid peptide homologous to the transmembrane envelope protein of retroviruses: evidence for a primary role played by monocytes.

CKS-17, a synthetic amino acid peptide homologous to a highly conserved region of retroviral transmembrane protein exerts a suppressive action on staphylococcal enterotoxin A (SEA)-induced the production of IFN-gamma by human peripheral blood mononuclear cells (PBMC) (Ogasawara et al., J. Immunol. 141, 615, 1988). This action has been shown in the present study to be preceded by dramatic clustering of PBMC. Clusters appear within 3 hr of exposure of PBMC to CKS-17; they are dose dependent, inhibited by cycloheximide, and require a temperature of 37 degrees C. The cells in the clusters are predominantly monocytes. Although it has been previously shown that CKS-17 inhibits monocyte-mediated killing by inactivating IL-1 (Kleinerman et al., J. Immunol. 139, 2329, 1987) and production of IL-2 by murine thymoma cells treated with IL-1 (Gottlieb et al., J. Immunol. 142, 4321, 1989), in the present study we show that IL-1 does not prevent clustering of PBMC by CKS-17. Using CKS-17 and highly purified monocytes or lymphocytes, profound alterations occur only with monocytes, as revealed by light or electron microscopy. SEA- or staphylococcal enterotoxin B-induced production of IFN-gamma is inhibited when highly purified monocytes pretreated with CKS-17 are cocultured with highly purified T lymphocytes. Thus, CKS-17 induces dramatic clustering of cells apparently by inducing alterations of monocytes but not lymphocytes, suggesting that CKS-17 may interfere with the capacity of monocytes to facilitate production of IFN-gamma by T lymphocytes.

Amino Acid Sequence↗

Enrichment of murine bone marrow natural suppressor activity in the fraction of hematopoietic progenitors with interleukin 3 receptor-associated antigen.

Natural suppressor (NS) activity has been identified in several sites of active hematopoiesis. In this study we characterized NS activity in murine bone marrow (BM) using monoclonal antibodies (mAbs) to interleukin 3 (IL-3) receptor-associated antigen (IL-3RAA) and various cytokines that exert a strong influence on hematopoiesis or lymphocyte interaction. NS activity of BM cells of relatively low density was enhanced by IL-3 or granulocyte-macrophage colony-stimulating factor (GM-CSF). When the BM cells were separated into IL-3RAA+ cells and IL-3RAA- cells, the IL-3RAA+ cells demonstrated potent NS activity, whereas IL-3RAA- cells had either no or weak NS activity. The IL-3RAA+ cells showed non-T- and non-B-cell phenotype and had high affinity to wheat germ agglutinin (WGA), a marker for hematopoietic progenitors. In assays for hematopoietic activity, it appeared that the early differentiating progenitors (day 8 spleen colony-forming units [CFU-S], granulocyte-macrophage colony-forming units [CFU-GM]) were enriched in the IL-3RAA+ cell population, whereas more immature multipotent progenitors (day 12 CFU-S, granulocyte erythrocyte macrophage megakaryocyte colony-forming units [CFU-GEMM]) were contained in the IL-3RAA- cell population. Both suppressor cells and IL-3RAA+ cells spontaneously developed from the IL-3RAA- cell population. These findings suggest that NS cells in murine BM are early hematopoietic progenitors and are probably committed to the myeloid lineage. Hybridoma cells established between the IL-3RAA+ cells and BW5147 cells produced suppressor factor(s). This finding suggests that the NS cells produce soluble mediator(s) that may be responsible for their suppressive action.

Animals↗

In vitro immunomodulation and in vivo immunotherapy of retrovirus-induced immunosuppression.

Early studies with the Gross passage A leukemia virus demonstrated that retroviral infection suppresses cellular and humoral immune responses. In extensive studies of the feline leukemia (FeLV) virus, which can induce profound immunodeficiency disease, are generative anemia and lymphoid, myeloid and erythroid neoplasia, the immunosuppressive effects of this retrovirus could be attributed to the actions of the retroviral envelope protein p15E. We found that a highly conserved, synthetic 17 amino acid peptide synthesized by Cianciolo and co-workers that is homologous to the hydrophilic portion of the otherwise hydrophobic transmembrane envelope protein can suppress polyclonal activation of B-cells, impair production of gamma- and alpha-interferon, inhibit production of interleukin-2, inhibit expression of IL-2 receptors, and suppress responses of cytotoxic lymphocytes. In analyses with inactivated preparations of the human immunodeficiency virus, with Pahwa et al. we demonstrated that purified non-infectious retrovirus and also retroviral proteins, in particular gp120, appeared to produce some of the immunosuppressive properties of HIV, particularly suppression of B-cell activation in response to known B-cell stimulants irrespective of T-cell influence, suppression of T-helper cell functions essential to B-lymphocyte responsiveness, and impaired function of immunoglobulin-secreting cells. Other investigators have also reported strong immunosuppressive or immunostimulatory influences for components of the HIV retrovirus and also gp120 through yet poorly elucidated but certainly complex actions on both T- and B-lymphocyte-mediated immune functions.

Animals↗

[Antibody-producing ability of mononuclear cells from human tonsils: a consideration for immunological pathogenesis and operative indication of recurrent tonsillitis].

Antibody-production of mononuclear cells from human tonsils was examined by enzyme-linked immunosorbent assays in the culture system following polyclonal stimulations with pokeweed mitogen. The results were summarized and discussed as follows; 1) Age-related changes were not observed. 2) Antibody-producing ability in the intermittent phase of recurrent tonsillitis was significantly impaired. 3) Antibody-production in the recrudescent phase of recurrent tonsillitis was increased and might be reactivated probably by stimuli of the recent infection in vivo. 4) It was suggested that the recurrent tonsillitis was due to the low antibody-production ability of tonsillar mononuclear cells. 5) These results gave an immunological basis on the operative indication of recurrent tonsillitis.

Adolescent↗

[A glomus tumor of the trachea--a case report].

We present a 61-year-old man with a glomus tumor of the trachea. To our knowledge, this is the 6th reported case in the literature. The patient was asymptomatic, and bronchoscopic examination revealed a highly vascular polypoid mass about 6.0 cm above the bifurcation of the trachea. We undertook a sleeve resection of the trachea with end to end anastomosis through a right posterolateral thoracotomy. The patient's postoperative course was uneventful. The tumor was 1.2 cm in diameter and was composed of glomus cells, which resembled to modified smooth muscle cells, ultrastructurally.

Glomus Tumor↗

[Studies on right ventricular performance in surgical treatment for primary lung cancer--especially in changes of right ventricular ejection fraction in acute phase].

In surgical treatment for lung cancer, increase of right ventricular afterload associated with decrease of pulmonary vascular bed after lung resection is serious problem in acute phase. Intensive care is necessary for the patients having major surgery, especially in aged. Until now, we have studied on pre and post operative extravascular lung water (EVLW) in addition to measurement of conventional hemodynamic changes on right ventricular afterload. And then, we have discussed when we should start to care intensively for degradation of compensatory hyperdynamic change. According to percent exchanges (delta %) between pre and post operative measurement, we established following criterion 1) delta %HR greater than 150% 2) delta SVI less than 60% 3) delta % PARI greater than 200% 4) observed postoperative EVLW greater than predicted postoperative EVLW. Furthermore, we investigated the correlation with these criterion and RVEF in 14 patients using a rapid computerized thermodilution method on right cardiac function in acute phase. We found that 1) changes of RVEF seem to depend on endsystolic volume 2) RVEF decreased just after operation and then slowly increased until 48 hours 3) PARI elevated to 194 +/- 117% in average at 6 to 24 hours showing significant inversed linear correlation to changes of RVEF (R = -0.54). The patients having prolonged elevation of PARI showed degradation of cardiac output and RVEF. RVSWI showed compensative acceleration gradually with increased right ventricular afterload.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

[Simplified prediction of postoperative lung function by plain chest roentgenogram in patients with primary lung cancer--in correlation to postoperative respiratory complications].

For the purpose of simplification of prediction of postoperative lung function, we studied to predict lung function by analizing the frontal and lateral view of chest plain roentgenogram and investigate the correlation to respiratory complication on 111 patients with lung cancer. According to TNM classification of lung cancer, prediction was performed as follows. Predicted postoperative lung function = [(42-number of resected subsegments)/(42-number of occupied subsegments)] x preoperative VC or FEV1.0. In this formula, 42 was the number of functioning subsegments of whole lung (right: 22, left: 20), and then preoperative occupied subsegments was ordered by T factor, where T1 lesion in lung field was prescribed as 1 subsegment and T2 was more than 2 subsegments respectively in plain chest roentgenogram. And also, on the patients having hilar lesions, it was required to calculate the number of subsegments in atelectasis, peripheral obstructive pneumonia and/or partial emphysematous change due to intrabronchial lesions. There was uniformly positive correlations in VC (R = 0.7949) and FEV1.0 (R = 0.8235) of the patients studied respectively. The patients having pneumonectomy showed tendency of over estimation, on the other hand, the patients having resection of a few segments showed under estimation. To predict the postoperative respiratory condition, we calculated the predicted post-operative %VC and %FEV1.0 for predicted preoperative normal VC and FEV1.0. Above the al, we tried to investigate the correlation with predicted postoperative %VC, %FEV1.0 and postoperative respiratory complications.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Studies on increased sputum after bronchoplastic surgery for lung cancer in correlation with hemodynamics and extravascular lung water].

In surgical treatment for lung cancer, large amount of intrabronchial sputum and secretion was occasionally encountered in patients, especially underwent bronchoplastic surgery. It has been discussed as causes which was affected from transection of pulmonary branches of vagal nerve and bronchial arteries. So we have measured pre- and postoperative hemodynamics and extravascular lung water (EVLW) at the bedside, as to investigate the correlation with formyl sputum and increase of right ventricular afterload which was presumed by lung resection and bronchoplasty. Respiratory complication was classified by the incidence of forced bronchial toilet in acute phase. Seven patients were investigated. Three of seven showed white sputum which was difficult to excrete. EVLW was 9.91 +/- 2.12 ml/kg in average before operation and then showed a tendency to increase within 102 +/- 29% of percent exchange (delta %) between pre- and postoperation. As to comparison of delta % of EVLW in acute period, the patients underwent bronchoplastic surgery showed a tendency to increase rather than the patients underwent lobectomy. According to former studies, we found the uniform correlation R = 0.7075 between observed EVLW and predicted postoperative EVLW estimated by prediction of residual functioning lung volume before operation. And also, the patients showing postoperative EVLW more than predicted EVLW before operation had a tendency to increase amount of sputum. Furthermore, concerning hemodynamic changes, pulmonary arterial resistance index (PARI) elevated to 196 +/- 165% (83-515%) in average associated with increased right ventricular stroke work index (RVSWI) as a result of compensatory changes for increased right cardiac afterload.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Inhibition of murine cytotoxic T lymphocyte activity by a synthetic retroviral peptide and abrogation of this activity by IL.

A synthetic 17 amino acid peptide (CKS-17) homologous to a highly conserved region of human and animal retroviral transmembrane proteins was investigated for its influence on the generation of murine alloantigen-specific CTL activity in vitro. CKS-17 coupled to a carrier protein, BSA or human serum albumin, inhibited the generation of anti-allo CTL in a dose-dependent manner. Controls consisting of BSA and human serum albumin, which had undergone the coupling procedure or neurotensin, an unrelated peptide, coupled to BSA in an identical manner as CKS-17 showed no such inhibitory action. The suppression was not restricted to the Ag specificity of the CTL activity. CKS-17 exerted inhibitory effects on the early afferent phase of CTL induction. Kinetic studies showed that the suppressive activity occurred when CKS-17 was introduced to the immunologically stimulating culture concomitant with or up to 48 h after initiation of culture. Analysis of the frequency of CTL precursor cells using limiting-dilution assays revealed that CKS-17 did act to reduce the number of precursor cells. Abrogation of the inhibition of CTL activity was observed when IL-2 was introduced to the culture together with the stimulator cells. Other lymphokines, such as IL-4, exerted a similar influence to counteract this suppression.

Animals↗

Partial purification of a growth factor synthesized by a rat hepatoma cell line established in serum-free medium.

A rat hepatoma cell line was established from primary culture using RPMI 1640 without supplements. Hepatomas were induced in rats by 0.06% 3'-methyl-4-dimethylaminoazobenzene. An established cell line, FF101, has been maintained as a monolayer for longer than 34 months and subcultured for 42 passages. The population-doubling time was 78 h. The modal chromosome number was 66. FF101 was transplantable, and morphological examination of the transplanted tumors revealed a mixed type of hepatocellular and cholangiocellular carcinoma. FF101 retained the ability to express tyrosine aminotransferase and glucose-6-phosphatase. Also, FF101 synthesized alpha-fetoprotein. FF101-conditioned medium stimulated DNA synthesis and proliferation of several cell lines such as AH66, K562, and BALB/c3T3. The growth-promoting activity of FF101-conditioned medium was abolished by protease, dithiothreitol, acidic treatment, and heating. Gel filtration of conditioned medium on Sephacryl S-200 disclosed the growth-promoting activity at the molecular size of approximately 60,000 Da, and the isoelectric point (pI) was between 5.5 and 6.5. These results suggest that FF101 synthesizes a novel growth factor which has little specificity in both species and organs.

Animals↗

Mechanism of target cell lysis by cytotoxic T lymphocytes (CTL) employing RSV-induced H-2 congenic and recombinant mouse tumor cells: demonstration of soluble mediators released from H-2-restricted CTL clones.

The secretion and the specificity of cytotoxic mediators from H-2-restricted cytotoxic T lymphocytes (CTL) were examined using non-virus-producing target tumor cells induced by the Schmidt-Ruppin strain of Rous sarcoma virus (SR-RSV) in B10 congenic and recombinant mice. By using rat concanavalin A supernatant, two H-2-restricted CTL clones were established from cytotoxic effector cells of B10.A(5R) mice primed with SR-RSV-induced syngeneic tumor Cell-free supernatants from the H-2-restricted CTL clones cocultured with syngeneic tumor cells had selectively high cytotoxic activity for syngeneic and H-2-compatible tumor cells, but not for H-2-incompatible tumor cells. YAC-1 cells, and B10.A(5R) blasts as defined in the 5-hr 51Cr-release assay. The cytotoxic activity was detected in the cell-free supernatants from the CTL clones cocultured with the CTL-sensitive syngeneic and H-2-compatible tumor cells, but not with the CTL-insensitive tumor cells and YAC-1 cells. The cytotoxic activity of the cell-free supernatant could be adsorbed by the syngeneic tumor cells, but not by YAC-1 and L(s) cells. Thus, the H-2-restricted CTL clones against SR-RSV-induced tumor cells were capable of releasing cytotoxic mediators by coculturing with syngeneic or H-2-compatible tumor cells, and the cytotoxic mediators showed a certain H-2-restricted manner in killing the target cells. These results suggest that the lysis of RSV-induced tumor cells by H-2-restricted CTL can at least in part be mediated by cytotoxic factors.

Adsorption↗