Biomedical subjects

T Horikawa

Publications and source records attributed to T Horikawa.

At least 37 records · Page 2Linked to original sources

PubMed2000

Association of latent membrane protein 1 and matrix metalloproteinase 9 with metastasis in nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a highly metastatic carcinoma whose consistent association with Epstein-Barr virus (EBV) has been established. Latent membrane protein 1 (LMP1), an EBV membrane protein expressed in latent infection, is considered to be the EBV oncoprotein. Matrix metalloproteinase 9 (MMP9), one of the MMP families, degrades Type IV collagen, a major component of extracellular matrix and is believed to be crucial for cancer invasion and metastasis. Although MMP9 is reported to be expressed in a variety of cancers, no reports concerning NPC have been published to date to the authors' knowledge. Recently, the authors have shown that LMP1 induces MMP9 in vitro cell line, which suggests the possibility of a mechanism in which LMP1 of EBV contributes to the metastasis and tumorigenesis of NPC by the induction of MMP9. METHODS: The expressions of LMP1 and MMP9 were immunohistochemically examined in 38 NPC sections, and the relation of these proteins were statistically analyzed. The authors also analyzed the associations of these proteins with clinical features. RESULTS: Both LMP1 and MMP9 proteins were predominantly immunolocalized in cancer nests. The expression of MMP9 showed a significant positive correlation with the expression of LMP1 (r = 0.75; P < 0.0001). Also, the expression of MMP9 correlated with lymph node metastasis (P = 0. 0004). CONCLUSIONS: The results suggest that the induction of MMP9 by LMP1 contributes to the metastatic potential of NPC.

Adaptor Proteins, Signal Transducing↗

PubMed2000

Transformation of Madin-Darby canine kidney (MDCK) epithelial cells by Epstein-Barr virus latent membrane protein 1 (LMP1) induces expression of Ets1 and invasive growth.

The Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP1) has a significant role in initiating EBV-associated lymphoproliferative disease and EBV-related malignancies. In view of clinical features related to the type of EBV latency, LMP1 may influence invasiveness of EBV associated tumors categorized as types II and III as represented on nasopharyngeal carcinoma (NPC). To screen for genes associated with invasion of epithelial cells transformed by LMP1, Madin-Darby canine kidney (MDCK) epithelial cells were transformed by LMP1. Stable transfection of a LMP1 gene into MDCK cells induced morphological change from cobblestone to a long spindle-shape, reduced cell-cell adhesion and caused high cell motility. Parental MDCK cells, which form spherical cysts in three-dimensional collagen gel matrix, form branching tubules following exposure to hepatocyte growth factor (HGF). MDCK cells transformed by LMP1 showed invasive growth to form branching tubules into collagen gel without HGF-treatment. mRNA differential display and Northern hybridization identified plasminogen activator inhibitor-1 (PAI-1), urokinase type plasminogen activator (uPA) and ets1 as genes upregulated during transformation by LMP1. Expression of a dominant negative type of Etsl in LMP1-transformed cells downregulated uPA expression and cell motility. Deletion of LMP1 cytoplasmic carboxy-terminal activating region 1 (CTAR1) domain abolished transformation, but a deletion mutant lacking CTAR2 domain still retained transforming and uPA-inducing ability. Expression of Ets1 was immunolocalized in tumor cells of NPC tissue which frequently express LMP1. Taken together, it is suggested that LMP1 induces expression of Ets1 which may contribute to invasion of NPC by stimulating cell motility and uPA expression.

Animals↗

PubMed2000

Roles for the protein tyrosine phosphatase SHP-2 in cytoskeletal organization, cell adhesion and cell migration revealed by overexpression of a dominant negative mutant.

SHP-2, a SRC homology 2 domain-containing protein tyrosine phosphatase, mediates activation of Ras and mitogen-activated protein kinase by various mitogens and cell adhesion. Inhibition of endogenous SHP-2 by overexpression of a catalytically inactive (dominant negative) mutant in Chinese hamster ovary cells or Rat-1 fibroblasts has now been shown to induce a marked change in cell morphology (from elongated to less polarized) that is accompanied by substantial increases in the numbers of actin stress fibers and focal adhesion contacts. Overexpression of the SHP-2 mutant also increased the strength of cell-substratum adhesion and resulted in hyperphosphorylation of SHPS-1, a substrate of SHP-2 that contributes to cell adhesion-induced signaling. Inhibition of SHP-2 also markedly increased the rate of cell attachment to and cell spreading on extracellular matrix proteins such as fibronectin and vitronectin, effects that were accompanied by enhancement of adhesion-induced tyrosine phosphorylation of paxillin and p130Cas. In addition, cell migration mediated by fibronectin or vitronectin, but not that induced by insulin, was impaired by overexpression of the SHP-2 mutant. These results suggest that SHP-2 plays an important role in the control of cell shape by contributing to cytoskeletal organization, and that it is an important regulator of integrin-mediated cell adhesion, spreading, and migration as well as of tyrosine phosphorylation of focal adhesion contact-associated proteins.

Animals↗

PubMed2000

Deficient Th1-type immune responses via impaired CD28 signaling in ultraviolet B-induced systemic immunosuppression and the restorative effect of IL-12.

A single large dose (15 kJ/m(2)) of UVB-irradiation induces systemic immunosuppression and tolerance. We previously reported that IL-12 promotes the accessory cell function of epidermal Langerhans cells. In this study we have examined whether IL-12-pretreated antigen-presenting cells (APC) could restore the diminished T-cell response in mice irradiated with a single large dose of UVB. Spleen cells from UVB-irradiated mice showed reduced IFN-gamma production in a hapten-specific response but the function of APC in non-exposed skin of UVB-irradiated mice was not impaired. The pretreatment of APC with IL-12 did not restore the impaired IFN-gamma production by T cells from UVB-irradiated mice. Neither IL-10 nor TGF-beta was found to be involved in the suppression. Instead, we observed that anti-CD3 mAb-induced IFN-gamma production by T cells from UVB-irradiated mice was not augmented in the presence of anti-CD28 mAb, whereas IL-4 production was enhanced by the addition of anti-CD28 mAb. Furthermore, the reduced IFN-gamma production by T cells from UVB-irradiated mice in response to antigen plus APC could be restored by adding IL-12 to the culture. Our results thus indicate that UVB-induced systemic immunosuppression involves impaired Th1-type responses of T lymphocytes through CD28 stimulation, and that IL-12 compensates for the impaired CD28 costimulatory signaling in T cells resulting in the restoration of Th1-type responses.

Animals↗

PubMed2000

Small Gtpase rab3A is associated with melanosomes in melanoma cells.

Rab3A is a small guanosine triphosphate (GTP)-binding protein that has been recently implicated in intracellular vesicle transport and the secretion of neurotransmitters in neuronal cells. We demonstrate here that Rab3A is associated with melanosomes in pigment cells. Rab3A as well as Rabphilin3A, a putative target protein of Rab3A, were detected in the melanosome fraction, purified from B16 murine melanoma cells by sucrose density gradient ultracentrifugation. In contrast, Rab GDP dissociation inhibitor (GDI), a GDP/GTP exchange protein for Rab3A, was found in the cytosol fraction. Further studies using confocal laser scanning microscopy and immunoelectron microscopy revealed that immunoreactive Rab3A is localized in conjunction with the melanosomal membrane. These results suggest the possibility of involvement of Rab3A-Rabphilin3A complex, regulated by Rab GDI, in the intracellular transport of melanosomes in pigment cells.

Adaptor Proteins, Signal Transducing↗

PubMed2000

Elimination of CD4(+) T cells enhances anti-tumor effect of locally secreted interleukin-12 on B16 mouse melanoma and induces vitiligo-like coat color alteration.

CD4(+) T cells have been reported to suppress immunity against cancer in certain animal models. In this study, we investigated the role of CD4(+) T cells in the anti-tumor immune response when interleukin-12-producing melanoma cells are inoculated in mice. We found that interleukin-12-transfected B16 melanoma showed retarded tumor growth in syngeneic mice; however, all the mice developed tumors eventually. In vivo depletion of CD4(+) T cells led to complete regression of B16/interleukin-12 tumors in 12 of 20 mice (60%). Immunohistochemical analyses revealed that a number of CD8(+) T cells accumulated in close proximity to the B16/interleukin-12 tumors in the CD4(+) T cell-depleted mice, whereas CD8(+) T cells were only scarcely observed at the periphery of the tumors in control immunocompetent mice. Furthermore, 10 of 20 mice treated with both B16/interleukin-12 inoculation and CD4(+) T cell depletion exhibited vitiligo-like coat color alteration. B16/interleukin-12 tumors completely regressed in all the mice with vitiligo. Histologic examination showed that CD8(+) lymphocytes accumulated around the hair bulbs of mice with vitiligo, but not in those without vitiligo. These results suggest that CD4(+) T cells have an inhibitory effect on tumor rejection by suppressing cytotoxic CD8(+) T cells in this melanoma loading model with local interleukin-12 secretion. To investigate the mechanism of enhanced anti-tumor effects by CD4(+) T cell depletion, we examined the T helper type 1/2 cytokine profile in the tumor draining lymph nodes of B16/interleukin-12-bearing mice with or without CD4(+) T cell depletion using the reverse transcription-polymerase chain reaction method. We found that CD4(+) T cell depletion eliminated T helper type 2 cells and resulted in a T helper type 1-dominant cytokine profile in tumor draining lymph nodes. We emphasize that this T helper type 1-dominant cytokine profile may generate further activated CD8(+) T cells against B16 melanoma cells, lead B16/interleukin-12 to regress, and result in the destruction of the melanocytes in hair bulbs due to cross-antigenicity between both cell types. This mouse model not only demonstrates the depletion of CD4(+) T cells as a useful strategy for cancer gene therapy with interleukin-12 but also provides a model for human melanoma-associated vitiligo.J Invest Dermatol 115:1059-1064 2000

Animals↗

PubMed2000

Mercury-induced nummular dermatitis.

We report 2 cases of relapsing nummular dermatitis according to mercury sensitivity, which was confirmed by patch testing. Removal of the amalgam from dental metal alloys markedly improved their skin eruptions. One of the patients, a dentist, experienced exacerbation of the eruptions on his lower legs after handling dental amalgam. Hypersensitivity to haptens such as metals is possibly involved in, at least in some patients, the pathogenesis of nummular dermatitis.

Adult↗

PubMed2000

Improvement of solubility and oral bioavailability of rutin by complexation with 2-hydroxypropyl-beta-cyclodextrin.

The object of this study was to enhance the solubility, dissolution rate, and oral bioavailability of rutin by complexation with 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CyD). The interaction of rutin with cyclodextrins (CyDs) was evaluated by the solubility, and ultraviolet (UV) and circular dichroism (CD) spectrophotometries. The chemical and enzymatic stability of rutin was examined in an alkaline buffer solution and in rat small intestinal homogenates, respectively. Dissolution rates of rutin and its CyD complexes were measured by the dispersed amount method. In vivo absorption studies of rutin after oral administration via conventional tablet containing rutin alone or its beta-CyD complexes was performed on beagle dogs. The stability constants calculated from the phase solubility method increased in the order of HP-gamma-CyD < G2-beta-CyD < beta-CyD < HP-beta-CyD. Spectroscopic studies also revealed that HP-beta-CyD and beta-CyD formed a relatively more stable inclusion complex with rutin. The dissolution rates of rutin increased by the complexation with CyDs in the order of rutin alone < HP-beta-CyD < or = beta-CyD. HP-beta-CyD inhibited the hydrolysis of rutin in the alkaline buffer solution and the small intestinal homogenates of rats, suggesting that HP-beta-CyD may stabilize rutin in a gastrointestinal tract after oral administration. When the tablet containing rutin or its beta-CyD complexes was administered to beagle dogs, the plasma levels of homovanillic acid (HVA) (a major stable metabolite of rutin) after oral administration of HP-beta-CyD complex were much higher than in either that of rutin alone or in its beta-CyD complex. The in vivo absorption study suggests that HP-beta-CyD increased the oral bioavailability of rutin from the gastrointestinal tracts of beagle dogs because of the increase in solubility, faster dissolution rate, and gastrointestinal stability. HP-beta-CyD has a significant advantage with respect to providing high aqueous solubility while maintaining a lack of toxicity in oral pharmaceutical preparations of rutin.

2-Hydroxypropyl-beta-cyclodextrin↗

PubMed2000

Heterozygous HPS1 mutations in a case of Hermansky-Pudlak syndrome with giant melanosomes.

We report a Japanese man with Hermansky-Pudlak syndrome, having oculocutaneous albinism with a bleeding diathesis. Gene analysis of the patient's peripheral blood cells revealed that he was a compound heterozygote for HPS1 gene mutations. One of the mutations was a novel frameshift mutation at codon 321 (a G insertion) in exon 11 ( approximately 962-963insG), and the other was a 5' splice-junction mutation of IVS5 (IVS5 + 5G-->A). The content of eumelanin in the patient's hairs was significantly reduced. Histological analysis using light and electron microscopy revealed that melanocytes in the patient's epidermis contained an appreciable number of giant melanosomes. Cultured melanocytes from the patient's skin also contained giant melanosomes. Our finding of mutations in the HPS1 gene in relation to abnormalities in melanosome morphology and melanin production shed light on the role and function of the HPS1 gene product in the synthesis of melanosomes and melanin pigment.

Adult↗

PubMed2000

An efficient synthesis of 5-bromo-2-methoxy-6-methylaminopyridine-3carboxylic acid.

An efficient synthesis of 5-bromo-2-methoxy-6-methylaminopyridine-3-carboxylic acid (1), a carboxylic acid moiety of a potent dopamine D2 and D3 and serotonin-3 (5-HT3) receptors antagonist, (R)-5-bromo-N-(1-ethyl-4-methylhexahydro-1 ,4-diazepin-6-yl)-2-methoxy-6-methylaminopyridine-3-carboxamide, is described. Reaction of methyl 2,6-difluoropyridine-3-carboxylate (12) with methylamine in EtOH at -25 degrees C gave a mixture of methyl 2-fluoro-6-methylaminopyridine-3-carboxylate (13) and the regioisomer 14 in a ratio of 57 : 43. On the other hand, reaction of 12 and methyl 2,6-dichloropyridine-3-carboxylate (16) with sodium methoxide in tetrahydrofuran (THF) and CH2Cl2 provided the 2-methoxypyridine-3-carboxylic esters 20 and 23, respectively, as main products. Similar reaction of 16 in N,N-dimethylformamide (DMF) and MeOH proved to be highly regioselective for the 6-position. A much greater regioselectivity for substitution at the 6-position (>97%) was observed when 16 was treated with 4-methylbenzenethiolate anion in DMF (quantitative yield). After methoxylation of methyl 2-chloro-6-(4-methylbenzenethio)pyridine-3-carboxylate (25b) and successive oxidation of the 6-benzenethio moiety, nucleophilic substitution of the sulfoxide derivative 28 with methylamine gave the 6-methylamino derivative 8. Finally, bromination of 8 and alkaline hydrolysis produced the desired product 1 in an overall yield of 67%.

Dopamine Antagonists↗

PubMed2000

Demonstration of phase-3 and phase-4 retrograde block in a second concealed accessory pathway after an initial successful radiofrequency ablation of a 'normal' concealed accessory pathway.

We report a patient with concealed Wolff-Parkinson-White syndrome who, following catheter ablation, demonstrated phase-3 and phase-4 retrograde block in a concealed accessory pathway. After an initial 'apparently successful' ablation, retrograde conduction was through the atrioventricular node during constant ventricular pacing. Ventricular extrastimulus testing was performed at a basic drive cycle length of 600 ms. Unexpectedly, ventricular extrastimuli at coupling intervals of 440-380 ms were conducted retrogradely over an accessory pathway, consistent with a phase-3 and phase-4 retrograde block in the accessory pathway. Residual accessory pathway conduction was eliminated in a single ablation session.

Catheter Ablation↗

PubMed2000

Diagnostic significance of the morphological change in the atrial electrogram during Para-Hisian pacing.

Para-Hisian pacing (PHP), a pacing method to differentiate between conduction occurring over an accessory pathway (AP) from that over the atrioventricular node (AVN), is assessed essentially by comparing the timing in the atrial electrogams. Morphological change in the atrial electrograms is often observed during PHP, but its significance has not been investigated. Prior to the catheter ablation procedure, PHP was performed in 52 patients with an AP and in 36 patients with AV nodal reentrant tachycardia (AVNRT). The morphological change in the atrial electrograms, which was retrospectively assessed between the His bundle and proximal right bundle branch (HB-RB) captured and non-captured beats, was identified in 15 of 52 patients with an AP and in 26 of 36 patients with AVNRT. The atrial electrogram in the 6 of these 15 AP patients changed its morphology without overlapping the ventricular electrogram. All 6 AP patients exhibited a PHP pattern with the presence of 2 retrograde conduction routes, an AP and the AVN. In the patients demonstrating no morphological change in the atrial electrogram, 33 of 37 AP patients and all 10 AVNRT patients had only one retrograde conduction route. Morphological change in the atrial electrogram without overlapping the ventricular electrogram seems to have diagnostic significance indicating the presence of both AP and AVN conduction.

Adolescent↗

PubMed2000

Antitumor effects on mouse melanoma elicited by local secretion of interleukin-12 and their enhancement by treatment with interleukin-18.

To investigate the mechanism of the antitumor effect of locally secreted interleukin-12 (IL-12), we introduced the IL-12 p35 and p40 cDNAs into mouse B16 melanoma cells. IL-12 gene-transfected B16 melanoma (B16/IL12) showed marked retardation of tumor growth when implanted subcutaneously into syngeneic mice. In these mice, depletion of not only Natural Killer (NK) cells but also CD8+ T cells diminished the antitumor effect of locally secreted IL-12. Immunohistochemical analysis showed that NK cells and macrophages accumulated more densely at the center and periphery of B16/IL12 tumors than that of parental B16 tumors, whereas CD4+ T cells and CD8+ T cells accumulated sparsely only at the periphery of both transfected and untransfected tumors. Systemic treatment with interleukin-18 (IL-18) markedly inhibited the growth of B16/IL12 but did not influence the tumor growth of parental B16 cells in vivo. These results suggest that local IL-12 secretion can retard the growth of B16 melanoma mediated primarily by NK cells and indirectly by CD8+ T cells and that its antitumor effect is augmented by systemic treatment with the novel cytokine IL-18.

Animals↗

PubMed2000

[A study of food-dependent exercise-induced anaphylaxis by analyzing the Japanese cases reported in the literature].

We surveyed and analyzed cases of food-dependent exercise-induced anaphylaxis (FDEIA) in the Japanese literature. We found 167 cases which were reported as FDEIA since 1983. Analyzing these case, following characteristic features were revealed: 1) Recent upward trend in the number of reports of FDEIA was noted. 2) Male cases were more frequent than female cases, while about half of the cases were teenager. 3) More than half of the cases were proved to be induced by wheat followed by shrimp. In those under 20, shrimp was the most popular cause rather than wheat. 4) Skin test and/or IgE RAST showed positive results in most cases, suggesting that the response itself is linked to type I allergic reaction. 5) About 40% of the patients had history of atopic disease, indicating that atopic condition may play some role in FDEIA. 6) All cases who performed provocation test with aspirin responded to food challenge and/or exercise in combination with aspirin. This indicates that aspirin plays a key provoking factor in FDEIA. 7) In 17 cases FDEIA attacks were observed during noon recess or physical education class after lunch at school. We stress here the importance of a nation-wide education to school teachers the potential danger of this disorder.

Adolescent↗

PubMed1999

Intermittent bundle branch blocks in a patient with uncommon-type atrioventricular nodal reentrant tachycardia and enhanced atrioventricular nodal conduction.

We report on a patient with uncommon-type atrioventricular (AV) nodal reentrant tachycardia with a short tachycardia cycle length (235-270 ms), in whom transient wide QRS tachycardia with both left bundle branch block and right bundle branch block aberrancy were followed by narrow QRS complexes. In addition, His-ventricular (H-V) block and a sudden prolongation of the H-V interval occurred during the tachycardia. As the determinant of these unusual findings, the possibility that the anterograde limb of the reentry circuit has an enhanced AV nodal conduction property is discussed, as is the clinical significance of this type of tachycardia.

Adult↗

PubMed1999

Effect of stress on atopic dermatitis: investigation in patients after the great hanshin earthquake.

BACKGROUND: Although a variety of factors are responsible for atopic dermatitis (AD), very little is known about the effect of stress caused by a natural disaster on AD symptoms. OBJECTIVE: This study was conducted to assess the effect of stress or trauma on the symptoms of AD. METHODS: One thousand four hundred fifty-seven patients with AD (745 males and 712 females), diagnosed by using the criteria of the Japanese Dermatology Association, were enrolled in the study. The data were obtained from a self-administered questionnaire given to patients with AD after experiencing the Great Hanshin Earthquake. The data were analyzed by multivariate logistic regression analysis. Patients were divided into 3 groups: area A, severe damage to buildings and houses; area B, mild damage; and control area, no damage. RESULTS: Exacerbation of skin symptoms was found in 38% and 34% of patients in areas A and B, respectively, whereas similar exacerbation was seen in 7% of control patients. Nine percent and 5% of the patients in areas A and B, respectively, showed improvement in their symptoms compared with 1% in the control group. The earthquake caused stress in 63% and 48% of the patients in areas A and B, respectively, but fewer patients felt stress in undamaged areas (19%). Multiple logistic regression analysis revealed that of the factors examined here, subjective distress is the factor that is most responsible for the exacerbation of skin symptoms (odds ratio, 2.98; 95% confidence interval, 2.25-3.96). CONCLUSION: The results presented here strongly suggest that stress caused by a natural disaster influences AD symptoms.

Adolescent↗

PubMed1999

Leiomyosarcoma of the larynx.

Tumours of smooth muscle origin are extremely rare in the upper aerodigestive tract. Immunohistochemical studies are helpful for accurate diagnosis of leiomyosarcoma. Here, we report a case of well-differentiated laryngeal leiomyosarcoma that showed rapid local growth but no metastasis. Diagnosis and treatment of leiomyosarcoma are discussed.

Actins↗

PubMed1999

Horizontal and vertical pigment spread into surrounding piebald epidermis and hair follicles after suction blister epidermal grafting.

Following the earlier description of Carnot and Deflandre in 1896, pigment spread phenomenon in mammals was investigated using immunogenetically marked melanocytes (Billingham and Silver, Quart. Rev. Biol. 1960 35: 1-40; Billingham and Silver, Ann. N.Y. Acad. Sci. 1963 100: 348-363). In spite of a number of similar studies on vitiligo lesions, detailed evaluation of pigment spread in piebald lesions has not been reported. To gain further insight into the pigment spread phenomenon in human skin, five piebald patients were studied, on whom suction blister epidermal grafting therapy onto piebald patches was performed. In the present study, pigmentation of all epidermal grafts from normally pigmented areas spread horizontally. It was also found that pre-existing white hairs in recipient sites became pigmented within 1 year after epidermal grafting. Immunofluorescence studies using melanocyte-specific antibody NKI/beteb revealed the newly induced presence of melanocytes in the newly pigmented hair follicles. Further, to study the possible mechanisms inhibiting melanocyte migration from normal skin into piebald lesions, epidermis was grafted from border zones (containing both normal and piebald skin) into the center of hypopigmented lesions. Melanocytes clearly migrated through the border zone of grafted epidermis into surrounding recipient hypopigmented sites.

Epidermis↗