Biomedical subjects

T Shin

Publications and source records attributed to T Shin.

At least 55 records · Page 3Linked to original sources

PubMed2000

Protein kinase C independent activation of inducible nitric oxide synthase by tumor necrosis factor-alpha in TM4 Sertoli cells.

To investigate the nitric oxide (NO) production and its signalling mechanism in TM4 Sertoli cells, the cells were treated with recombinant tumor necrosis factor-alpha (rTNF-alpha), recombinant interleukin-1 alpha (rIL-1alpha), or lipopolysaccharide (LPS), either alone or in combination with recombinant interferon-gamma (rIFN-gamma), and NO production was measured by using the Griess method. TM4 Sertoli cells produced a small amount of NO upon treatment with rIFN-gamma. The effect of rIFN-gamma was drastically increased by cotreatment with rTNF-alpha in a dose-dependent manner. However, combination of rIL-1alpha or LPS with rIFN-gamma did not synergize to activate cells. RIFN-gamma in combination with rTNF-alpha showed marked increase of the expression of iNOS protein. Protein kinase C inhibitors did not inhibit the production of NO induced by rIFN-gamma plus rTNF-alpha. These results suggest that the role of TNF-alpha is to provide TM4 Sertoli cells with the active cofactor for NO production and TNF-alpha-induced signaling for induction of NO synthesis is not dependent on protein kinase C activation.

Animals↗

PubMed2000

Correlative assessment of hemodynamic parameters obtained with T2*-weighted perfusion MR imaging and SPECT in symptomatic carotid artery occlusion.

BACKGROUND AND PURPOSE: Perfusion MR imaging and single-photon emission CT (SPECT) are commonly used to evaluate hemodynamic status in patients with symptomatic occlusive cerebrovascular disease. These techniques rely on different underlying physiological mechanisms, and the data may not correspond. We studied the relationship between hemodynamic parameters obtained with these two methods. METHODS: We performed perfusion MR imaging and SPECT in 10 patients with symptomatic unilateral internal carotid artery occlusion. Relative cerebral blood volume (rCBV) and uncorrected mean transit time (uMTT) were obtained with dynamic contrast-enhanced T2*-weighted MR imaging. Relative cerebral blood flow (rCBF) and vascular reserve capacity were measured with 99mTc-HMPAO SPECT; vascular reserve capacity was calculated by the difference in CBF before and after acetazolamide challenge. Ratios of these hemodynamic parameters between the affected and contralateral vascular territories were calculated and compared. RESULTS: Normal-to-increased CBV, prolonged uMTT, decreased CBF, and normal-to-diminished vascular reserve capacity were observed in the affected vascular territories. Reduction of vascular reserve capacity corresponded well with uMTT but not with CBF and CBV. CBF, CBV, and uMTT did not correspond to one another. CONCLUSION: uMTT is more sensitive than the other parameters in estimating vascular reserve capacity. The relationship between parameters obtained with perfusion MR imaging and SPECT should be considered in assessing the hemodynamic status of patients with symptomatic occlusive cerebrovascular disease.

Aged↗

PubMed2000

Enhanced expression of constitutive and inducible forms of nitric oxide synthase in autoimmune encephalomyelitis.

To elucidate the role of nitric oxide synthase (NOS) in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), we analyzed the expression of constitutive neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS) in the spinal cords of rats with EAE. We further examined the structural interaction between apoptotic cells and spinal cord cells including neurons and astrocytes, which are potent cell types of nitric oxide (NO) production in the brain. Western blot analysis showed that three forms of NOS significantly increased in the spinal cords of rats at the peak stage of EAE, while small amounts of these enzymes were identified in the spinal cords of rats without EAE. Immunohistochemical study showed that the expression of either nNOS or eNOS increased in the brain cells including neurons and astrocytes during the peak and recovery stages of EAE, while the expression of iNOS was found mainly in the inflammatory macrophages in the perivascular EAE lesions. Double labeling showed that apoptotic cells had intimate contacts with either neurons or astrocytes, which are major cell types to express nNOS and eNOS constitutively. Our results suggest that the three NOS may play an important role in the recovery of EAE.

Animals↗

PubMed2000

Immunohistochemical study of constitutive neuronal and inducible nitric oxide synthase in the central nervous system of goat with natural listeriosis.

The expression of both constitutive and inducible forms of nitric oxide synthase (NOS) was investigated by immunohistochemical staining of formalin-fixed paraffin-embedded sections in normal and Listeria monocytogenes-infected brains of goats. In normal control goats, a small number of neurons showed immunoreactivity of both iNOS and nNOS, and the number of iNOS-positive neurons was higher than the number of nNOS-positive neurons. In natural listeriosis, listeria antigens were easily immunostained in the inflammatory cells of microabscesses. In this lesion, the immunoreactivity of iNOS in neurons was more intense than the control, but nNOS was not. In microabscesses, nNOS was weakly visualized in macrophages and neutrophils, while iNOS was expressed in macrophages, but not in neutrophils. These findings suggest that normal caprine brain cells, including neurons, constitutively express iNOS and nNOS, and the expressions of these molecules is increased in Listeria monocytogenes infections. Furthermore, inflammatory cells, including macrophages, expressing both nNOS and iNOS may play important roles in the pathogenesis of bacterial meningoencephalitis in goat.

Animals↗

PubMed1999

Differential role of TNF-alpha and IFN-gamma in the brain of rats with chronic relapsing autoimmune encephalomyelitis.

To elucidate the mechanisms of relapses of the clinical signs in experimental autoimmune encephalomyelitis (EAE), the cytokine profile of chronic relapsing EAE (CR-EAE) in rats was determined by competitive polymerase chain reaction (PCR). By immunization with guinea pig spinal cord homogenate and treatment with low-dose cyclosporin A (CsA), rats developed two attacks of EAE with remission in between. Cytokine analysis revealed that the level of TNF-alpha mRNA increased at the first and second attacks with transient disappearance at the remission phase. In contrast, the level of IFN-gamma mRNA was suppressed at the first attack by CsA and peaked at the second attack. Intraventricular administration of IFN-gamma prior to onset of disease signs induced more relapses, or a severe lethal form. In addition, the intraventricular injection of TNF-alpha caused the persistence of the clinical signs. These findings suggest that TNF-alpha contributes to the first and second attacks of CR-EAE, while IFN-gamma is not required for the first attack but is closely related to the relapse of the disease. With regard to anti-inflammatory cytokines, the levels of both TGF-beta1 and IL-10 mRNA at the second attack were higher than those at the first attack. Taken together, differential involvement of TNF-alpha and IFN-gamma is closely associated with the clinical features of CR-EAE.

Animals↗

PubMed1999

Occipital screw pullout strength. A biomechanical investigation of occipital morphology.

STUDY DESIGN: A three-group design with consistent pullout strength measures. OBJECTIVES: To determine pullout strength of three fixation types (unicortical screws, bicortical screws, wires) and to investigate their correlation with respect to occipital morphology. SUMMARY OF BACKGROUND DATA: A secured, multidirectional occipitocervical fusion requires internal fixation. Devices secured at occipital protuberance were suggested to offer the greatest pullout strength because of this region's thickness. METHODS: Twelve fresh human cadaveric occiputs were sketched with a grid delineating 21 fixation sites. Each site was drilled and hand-tapped. Four specimens were instrumented with unicortical screws on one side of the midline and bicortical screws on the other. Another four were instrumented with bicortical screws and wires, and the remaining four were instrumented with unicortical screws and wires. Two points on each specimen were secured with identical fixation to examine side-to-side symmetry. An MTS materials testing apparatus (MTS Systems Corporation, Eden Prairie, MN) was used to displace the fixators. Pullout strengths at different anatomic locations were recorded. RESULTS: The greatest pullout strength was at the occipital protuberance for all fixation types. The bicortical pullout strength was 50% greater than unicortical. The wire pullout strength was not significantly different from that of the unicortical screw (P > 0.05). Seventy-eight percent of wires broke at 1100 N. Unicortical pullout strength at occipital protuberance was comparable with that of the bicortical screw at other locations. CONCLUSIONS: Unicortical screw fixation at occipital protuberance offers acceptable pullout strength without the potential complications of bicortical screws or wire fixation.

Biomechanical Phenomena↗

PubMed1999

Effects of stromal fibroblasts and fat cells and an environmental factor air exposure on invasion of laryngeal carcinoma (HEp-2) cells in a collagen gel invasion assay system.

OBJECTIVE: To clarify the invasion mechanism of laryngeal carcinoma (HEp-2) cells. DESIGN: Human HEp-2 cells were cultured on a collagen gel containing fibroblasts and/or fat cells. The HEp-2 cells were also treated with air exposure as the local environment of the laryngeal epithelial mucosa. A collagen gel invasion assay was conducted under these culture conditions. RESULTS: No invasion of HEp-2 cells was found in the stromal cell-free collagen gel, but a slight invasion was observed in the fibroblast-embedded gel. A deeper invasion of HEp-2 cells occurred in the fibroblast- and fat cell-coembedded gel and in the fibroblast-embedded gel with air exposure. The most extensive invasion of HEp-2 cells was observed under the fibroblast- and fat cell-coembedded gel in combination with air exposure. CONCLUSIONS: Fat cells and air exposure clearly increase the invasive effect of fibroblasts in squamous cell carcinoma of the larynx. The combined effect of these 3 factors (ie, fat cells, fibroblasts, and air exposure) plays a very important role in the invasive growth of the carcinoma cells. This observation suggests that both tumor cell-stromal cell interaction and tumor cell-local environmental factor interaction should be taken into account in an investigation of the invasive and proliferative mechanisms of laryngeal carcinoma.

Adipose Tissue↗

PubMed1999

Can secondary degeneration accelerate the formation of neurofibrillary tangles? A case of hemispheric infarction showing asymmetric degeneration of the substantia nigra, red nuclei, inferior olivary nuclei and dentate nuclei with concomitant changes of progressive supranuclear palsy.

A case of hemispheric infarction involving the territory of the right middle cerebral artery and the thalamus showed conspicuous asymmetric degeneration in the substantia nigra, red nuclei, inferior olivary nuclei and dentate nuclei with concomitant changes of progressive supranuclear palsy (PSP). The right substantia nigra and red nucleus showed loss of neurons and proliferation of astrocytes. The right olivary nucleus was hypertrophic, while the neuronal loss and astrocytosis in the dentate nucleus were predominant on the contralateral side. Modified Gallyas-Braak staining revealed the extensive distribution of neurofibrillary tangles (NFTs), threads and intraglial argyrophilic structures in the globus pallidus, subthalamic nuclei, cerebral cortex and dentate nuclei, as well as in the affected brain stem nuclei, with a distinct predominance on the affected side. In this case, the one-sided predominance of the extended degeneration in these brain stem and cerebellar areas is considered, in addition to the PSP changes, to be due to secondary retrograde degeneration via the nigrostriatal and dentato-rubro-thalamic pathways following the hemispheric infarction, and to also be the result of disruption of the dentato-olivary fiber connections. In addition, because of the predominant distribution of NFTs on the more degenerated side, it is surmised that the formation of NFTs may be accelerated by secondary degeneration.

Aged↗

PubMed1999

Expression and localization of protein kinase C theta isoform in mouse testis.

Protein kinase C (PKC) is encoded by a complex of a gene family, and its multiple isoforms are expressed in various mammalian tissues. The objective of this study was to investigate the expression and localization of a PKC theta isoform in mouse testis. PKC theta displays the highest homology to PKC delta, lacks the Ca2+-binding C2 domain and, thus, belongs to the subfamily of Ca2+-independent PKC enzymes which also includes the delta, epsilon, zeta and eta isoforms. We analyzed the PKC theta mRNA and protein by Northern blotting, in situ hybridization, and immunohistochemistry. In testes of normal mice, signals of PKC theta isoform expression were detected specifically in the interstitial cells of testes. The expression of PKC theta isoform was also detected in testes of germ cell-deficient W/W(v) mice. These results suggest that PKC theta isoform has the specific biological functions in the interstitial cells of testis.

Animals↗

PubMed1999

Pharyngeal branch of the vagus nerve carries intraepithelial afferent fibers in the cat pharynx: an elucidation of the origin and central and peripheral distribution of these components.

The presence of a sensory component in the pharyngeal branch of the vagus nerve (PhB), including its peripheral distribution and central projection, was studied by denervation and tracer experiments in the cat. The distribution of nerve fibers immunoreactive to protein gene product 9.5, a sensitive neuronal marker; calcitonin gene-related peptide; and substance P in the pharyngeal epithelium was analyzed in both intact animals and animals subjected to partial denervation by means of sectioning two of the three nerve trunks, the glossopharyngeal nerve, the superior laryngeal nerve, and the PhB, while leaving one intact. The results of this study show that the glossopharyngeal nerve and superior laryngeal nerve carry nerve fibers to the pharyngeal epithelium rostral and caudal to the middle level of the epiglottis, respectively, whereas the PhB carries nerve fibers to the mesopharyngeal epithelium. Tracer experiments, by applying wheat germ agglutinin-horseradish peroxidase to the PhB, demonstrated retrogradely labeled primary sensory neurons in the jugular ganglion and transganglionic labeling of terminals in the interstitial subnucleus of the nucleus of the tractus solitarius. These results indicate that the PhB contains a sensory component that originates from the jugular ganglion, innervates the mesopharyngeal epithelium, and projects to the nucleus of the tractus solitarius.

Afferent Pathways↗

PubMed1999

Histopathological analysis of apoptosis, and expression of p53, bcl-2, bax, and Ki-67 in laryngeal squamous cell carcinomas and dysplasia.

The growth of neoplasia is determined by the proliferation and loss of cells. The purpose of this study is to determine the frequency of apoptosis in laryngeal carcinomas and to examine its relationship to the pathological parameters, including ki-67 expression, and to expression of p53, bcl-2, and bax protein. The materials are 67 cases of laryngeal squamous cell carcinomas (SCCs) and 22 cases of squamous dysplasia using biopsy and surgery specimens. Apoptotic cells were determined by the modified TUNEL method. Expressions of p53, bcl-2, and bax, i.e. apoptosis-related genes, and ki-67, a proliferation marker, were immunohistochemically examined. The relationships between apoptosis and the clinicopathological findings were studied. The stage of the carcinoma was not related to the apoptotic index. The expression of p53 was frequently detectable in the advanced carcinomas with T3, T4 and N-positive. The apoptotic index was not significantly related to recurrence, metastasis or histological differentiation. Apoptosis occurred frequently in the cornified areas of well differentiated SCCs. The expressions of ki-67 observed in the poorly differentiated SCCs was significantly higher than that observed in the well differentiated SCCs (P< 0.01). The apoptotic index increased after irradiation in the carcinoma. No relationship was found between apoptotic index, ki-67 index, and expression of p53, bcl-2 and bax. The apoptotic index obtained form the SCCs was significantly higher than that obtained form squamous dysplasias (P < 0.05). Various apoptosis-related findings including p53 expression were observed in the advanced type of laryngeal SCCs, and apoptosis of the carcinoma was suggested to be controlled by complicated factors including bcl-2.

Adult↗

PubMed1999

An encyclopedia of mouse genes.

The laboratory mouse is the premier model system for studies of mammalian development due to the powerful classical genetic analysis possible (see also the Jackson Laboratory web site, http://www.jax.org/) and the ever-expanding collection of molecular tools. To enhance the utility of the mouse system, we initiated a program to generate a large database of expressed sequence tags (ESTs) that can provide rapid access to genes. Of particular significance was the possibility that cDNA libraries could be prepared from very early stages of development, a situation unrealized in human EST projects. We report here the development of a comprehensive database of ESTs for the mouse. The project, initiated in March 1996, has focused on 5' end sequences from directionally cloned, oligo-dT primed cDNA libraries. As of 23 October 1998, 352,040 sequences had been generated, annotated and deposited in dbEST, where they comprised 93% of the total ESTs available for mouse. EST data are versatile and have been applied to gene identification, comparative sequence analysis, comparative gene mapping and candidate disease gene identification, genome sequence annotation, microarray development and the development of gene-based map resources.

Animals↗

PubMed1999

Purification and characterization of a novel 5-oxoprolinase (without ATP-hydrolyzing activity) from Alcaligenes faecalis N-38A.

A novel type of 5-oxoprolinase was found in a cell extract of strain N-38A, which was later identified as Alcaligenes faecalis. The enzyme in the cell extract was purified to a homogeneous state with a yield of 16.6%. The molecular weight of the purified enzyme was estimated to be 47,000 by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, suggesting that the enzyme is a monomeric protein. The enzyme specifically catalyzed a decyclization of L-pyroglutamate without hydrolyzing ATP and also without any requirements for metal ions such as Mg2+ and K+. The optimal pH for the decyclization was 7.4. The reaction was reversible. The equilibrium constant of the reaction, Keq = [L-glutamate]/[L-pyroglutamate], was evaluated to be approximately 0. 035, which indicates that the reaction tends to form L-pyroglutamate. The amino-terminal amino acid sequence of the enzyme was H-Glu-Pro-Arg-Leu-Asp-Thr-Ser-Gln-Leu-Tyr-Ala-Asp-Val-His-Phe-. No protein with a similar sequence was found in the DNASIS database. Based on these data, it was strongly suggested that the enzyme described here is a novel type of 5-oxoprolinase.

Journal Article↗

PubMed1999

A case of ALK-positive large T-cell lymphoma expressing epithelial membrane antigen with favorable prognosis.

We describe the case of a 59-year-old woman with Ki-1-negative, T cell-type, anaplastic lymphoma kinase (ALK)-positive large cell lymphoma that was positive for epithelial membrane antigen. She was histopathologically diagnosed as having a metastatic undifferentiated carcinoma from a cervical lymph node biopsy. Clinical and radiographic studies revealed no other primary tumor. The patient underwent a left radical neck dissection. Surgically resected lymph nodes revealed an ALK1-positive large cell lymphoma. Thereafter the patient has had an unusually favorable prognosis.

Anaplastic Lymphoma Kinase↗

PubMed1999

Various patterns of perfusion-weighted MR imaging and MR angiographic findings in hyperacute ischemic stroke.

BACKGROUND AND PURPOSE: Various clinical subtypes of patients presenting with sudden-onset ischemic stroke have been recognized, but classification of those types is not simple. We identified various patterns of perfusion-weighted MR imaging and MR angiographic findings in hyperacute ischemic stroke with relation to clinical outcomes. METHODS: Twelve patients with symptoms of acute ischemic stroke due to middle cerebral artery occlusion underwent perfusion-weighted MR imaging and MR angiography within 6 hours after the onset of symptoms. Perfusion-weighted imaging was performed with a conventional dynamic contrast-enhanced T2*-weighted sequence, and cerebral blood volume (CBV) maps were then created. CBV maps and MR angiographic findings were compared with 99mTc-HMPAO brain SPECT scans, short-term outcomes, and follow-up imaging findings. RESULTS: The combined CBV and MR angiographic findings were classified into three patterns: arterial occlusion and decreased CBV (n = 8), arterial occlusion and increased CBV (n = 2), and no arterial occlusion and normal CBV (n = 2). These three patterns were strongly related to SPECT findings, short-term outcomes, and follow-up imaging findings. Perfusion on SPECT decreased markedly in the affected regions in all patients with the first pattern, decreased slightly in the second pattern, and was normal in the third pattern. Symptoms were not significantly changed at 24 hours after onset in any of the patients with the first pattern, but resolved completely in all patients with the latter two patterns. Follow-up imaging showed large infarctions in all patients with the first pattern. Initially, no infarction was seen in the second pattern, but watershed infarction developed later in one of these patients. CONCLUSION: Hyperacute ischemic stroke may be differentiated into three imaging patterns with different clinical outcomes. The combined use of perfusion-weighted MR imaging and MR angiography may play a substantial role in guiding the choice of treatment of this disease.

Acute Disease↗

PubMed1999

Loss of retinoblastoma protein expression in laryngeal squamous cell carcinoma.

Laryngeal carcinomas are among the most curable malignancies, but some of them show poor clinical outcomes with local recurrence or regional neck metastasis. Multiple genetic alterations of oncogenes and tumor suppressor genes are thought to occur in the development some tumors. The frequency, however, and the prognostic significance of these genes or gene products still remain unknown in laryngeal carcinoma. Epidemiologic data suggest that cigarette smoking is closely related to the development of these neoplasms; and human papillomavirus (HPV) infections have also been postulated to play a role in development of laryngeal tumor. The objective of this study was to investigate the presence of retinoblastoma protein (pRb), p53 protein, and HPV infection in laryngeal squamous cell carcinoma, as well as the correlation of these factors with clinicopathologic and carcinogenic factors. Tumors from 79 patients with primary laryngeal squamous cell carcinoma were studied. The expression of pRb was immunohistochemically assessed in 79 such tumors, and the expression of p53 was assessed in 76 tumors. HPV Type 16 expression was estimated by nonisotopic in situ hybridization in 78 cases. Follow-up periods ranged from 15 to 90 months. pRb was detected in 82% of the tumors and p53 in 43%. Lack of staining for pRb was significantly associated with a high-histologic grade (P<.05), high T classification (P<.05), recurrence (P<.05), and a relatively short disease-free interval (P<.01). p53 overexpression was observed frequently in heavy smokers with an average Brinkman index of more than 800, but it was not associated with any of the clinicopathologic factors studied. These findings suggest that tumors exhibiting loss of pRb expression have a more aggressive biologic behavior than do those that express pRb and that loss of pRb expression might predict clinical outcome in patients with primary laryngeal squamous cell carcinoma.

Aged↗

PubMed1998

An inhibitor of inducible nitric oxide synthase ameliorates experimental autoimmune myocarditis in Lewis rats.

We studied the effect of nitric oxide (NO) on experimental autoimmune myocarditis (EAC) in rats. We examined the role of inducible nitric oxide synthase (iNOS), an enzyme that produces NO, on hearts affected with EAC, by testing the effects of aminoguanidine (AG), a selective iNOS inhibitor, on the course of EAC. Western blotting detected iNOS in the affected cardiac tissues, but not in CFA immunized cases. Immunohistochemically, the majority of ED1+ macrophages in the EAC lesions were positive for iNOS and nitrotyrosine. A high dose of AG (200 mg/kg/day) significantly reduced the incidence of EAC (p < 0.05) and ameliorated the histological score for the cardiac inflammation (p < 0.01) compared with the low dose AG (100 mg/kg/day) and vehicle treated groups. The immunoblot analysis showed that a high dose of AG effectively suppressed iNOS in hearts affected with EAC. An iNOS band was barely detected in the high dose AG (200 mg/kg) treated group, while it was distinctively visualized in the vehicle and low dose AG (100 mg/kg) treated groups. These results suggest that iNOS is upregulated in EAC lesions and increased NO production plays an important role in the development of EAC. In addition, selective iNOS inhibitors may have a therapeutic role in treating certain autoimmune diseases including EAC.

Animals↗

PubMed1998

Cloning of a thermostable ascorbate oxidase gene from Acremonium sp. HI-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis.

A gene encoding a thermostable ascorbate oxidase (ASOM) was cloned from Acremonium sp. HI-25 and sequenced. The gene comprised 1709 bp and was interrupted by a single intron of 57 bp. ASOM consisted of 551 amino acids including a signal peptide with a molecular mass of 61200, and contained four histidine-rich regions with high sequence homology to the corresponding regions of other multicopper oxidases. The ASOM gene was expressed in Aspergillus nidulans under the Aspergillus oryzae Taka-amylase A gene promoter. The recombinant enzyme (An-ASOM) exhibited almost the same enzymatic properties as ASOM. The ASOM gene was mutated by site-directed mutagenesis with reference to the amino acid sequences of plant enzymes to generate enzymes with altered azide sensitivity. Site-directed mutagenesis at the trinuclear active copper site resulted in an increase in azide resistance; the Ala465Leu and Phe463Trp/Ala465Leu mutants exhibited approximately 10 and 20% increases in azide resistance, respectively.

Acremonium↗