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T Kuroki

Publications and source records attributed to T Kuroki.

At least 163 records · Page 9Linked to original sources

Isolation and characterization of a putative keratin-associated protein gene expressed in embryonic skin of mice.

Embryonic mouse skin undergoes substantial morphologic changes from 13 days post-coitus (dpc) to 16 dpc, i.e., from simple layers of epithelial cells and periderm at 13.5 dpc to almost fully differentiated stratified epithelium with the rudiments of hair follicles at 16.5 dpc. Using RNA differential display, we isolated a gene involved in the development of mouse epidermis. This gene, tentatively designated as 4C32, encodes 197 amino acids containing six direct repeats of 10 amino acids with the CQ motif. The repetitive structure with the CQ motif is seen in most keratin-associated protein families, which are known to be specifically expressed in hair follicles. 4C32 is expressed in the outermost layer of the embryonic epidermis at 15.5 and 16.5 dpc, and abruptly disappears at 17.5 dpc, suggesting that 4C32 is expressed in the periderm. The periderm is a superficial layer of embryonic epidermis, and is known to disappear at 17 dpc in mouse embryos. The 4C32 transcripts were also detected in the developing and matured tongue tissues and in the tail scale, but not at any stage in hair follicles.

Amino Acid Sequence↗

Cholesterol sulfate activates transcription of transglutaminase 1 gene in normal human keratinocytes.

Cholesterol sulfate and transglutaminase 1 are essential for the process of keratinization. Cholesterol sulfate is formed during keratinization and activates the eta isoform of protein kinase C. Transglutaminase 1 is a key enzyme for formation of the cornified envelope in terminally differentiated keratinocytes. In this study, we demonstrated that cholesterol sulfate acts as a transcriptional activator of the transglutaminase 1 gene in normal human keratinocytes. Growth of normal human keratinocytes was inhibited by cholesterol sulfate, but not by its parental cholesterol. Treatment of normal human keratinocytes with cholesterol sulfate induced activity of transglutaminase 1 in a dose- and time-dependent manner. Activation of transcription of transglutaminase 1 by cholesterol sulfate was demonstrated by northern blotting analysis, whereas that by cholesterol was not. In order to identify a cholesterol sulfate responsive region in the transglutaminase 1 gene, plasmids were constructed containing a luciferase reporter gene ligated to deletion fragments of the 5' upstream region of the tranglutaminase 1 gene and were transfected into normal human keratinocytes. Transfected cells were treated with cholesterol sulfate, the phorbol ester 12-O-tetradecanoylphorbol-13-acetate and a high concentration of Ca2+. Our results indicate that the responsive element(s) for cholesterol sulfate and phorbol ester is located upstream of the human transglutaminase 1 gene at a position(s) between -819 and -549, whereas the responsive element for Ca2+ is located at a position between -79 and -49.

5' Untranslated Regions↗

External-internal oblique reverse blood supply musculocutaneous flap for chest wall reconstruction.

A defect resulting from resection of advanced breast tumor can be quite large, posing a difficult reconstructive challenge. A significant number of such patients are found to have local recurrences after receiving beam radiation therapy and chemotherapy. The external oblique musculocutaneous flap is now considered a reliable option, with segmental blood supply from the posterior intercostal arteries. We report a 57-year-old man with local recurrence of breast tumor after external beam therapy. We faced a difficult challenge in performing reconstruction because of (1) several previous operations that used most of the usual options and (2) the absence of recipient vessels for microsurgery. We performed an external-internal oblique musculocutaneous flap with a reverse blood supply from the lateral branches of the inferior epigastric artery. Our use of a reverse-flow external-internal oblique flap supports the concept of rotating the muscles as a unit to encourage viability of the denervated and relatively devascularized component.

Arteries↗

Usefulness of scintigraphy with Tc-99m phytate for the diagnosis of alcoholic foamy degeneration.

PURPOSE: Alcoholic foamy degeneration (AFD) is a liver disease causing temporary hepatocyte dysfunction. The prognosis is usually good, but liver biopsy is needed for diagnosis. We report the usefulness of liver-spleen scintigraphy with the radiocolloid Tc-99m phytate for the diagnosis of AFD. PATIENTS AND METHODS: We used liver scintigraphy with Tc-99m phytate to study three patients with AFD diagnosed on the basis of findings from a liver biopsy. RESULTS: Liver-spleen scintigraphy showed hepatomegaly and splenomegaly, and bone marrow was visible, but radioisotope uptake by the liver was uniform. CONCLUSIONS: This pattern of scintigraphic findings is different from that reported for patients with alcoholic fatty livers or severe alcoholic hepatitis, and seems to be specific for AFD.

Adult↗

A case of cap polyposis investigated by scintigraphy with human serum albumin labeled with Tc-99m DTPA.

Cap polyposis is a rare intestinal disease that can be difficult to differentiate from inflammatory bowel disease. When cap polyposis is suspected, it is important to confirm protein loss. A 54-year-old woman who had been treated for ulcerative colitis for 7 years had severe hypoproteinemia. Scintigraphy with Tc-99m-labeled DTPA complexed with human serum albumin showed protein loss from the descending colon. Left hemicolectomy and sigmoid colectomy were performed. Cap polyposis was diagnosed on the basis of histologic findings from an operative specimen. The patient's diarrhea resolved after surgery and her hypoproteinemia improved. Scintigraphy with this label gave information helpful in the diagnosis of cap polyposis.

Colitis, Ulcerative↗

Prostaglandins in the stomach: an update.

Prostaglandins (PGs) are responsible for regulation of various physiologic activities in many tissues and organs, including the stomach. Recent studies have shown new crucial roles of PGs in the stomach. Activation of inflammatory cytokines and neutrophils may cause acute gastric mucosal lesions and recurrence of ulcers, which are induced by noxious stimuli such as nonsteroidal antiinflammatory drugs (NSAIDs), stress and Helicobacter pylori (H. pylori). These phenomena are PG-dependent because exogenous PGs reverse them. PG deficiency and H. pylori may worsen the quality of ulcer healing in terms of inflammatory responses, which are related to future ulcer recurrence. Neutrophils, monocytes/macrophages, and adhesion molecules are involved in the mechanism of recurrence caused by inflammatory cytokines. PGs accelerate ulcer healing, possibly via angiogenesis, epithelial cell proliferation, production of growth factors such as hepatocyte growth factor and transforming growth factor beta, reconstruction of extracellular matrices, and suppression of inflammatory cell infiltration, in addition to gastroprotective mechanisms. The PG synthase cyclooxygenase (COX) has two forms, COX-1 and COX-2. COX-2, but not COX-1, contributes to ulcer healing. Moreover, recent studies suggest the involvement of COX-2 in development of gastric and colon carcinoma. This may be linked to the chemopreventive effect of NSAIDs.

Humans↗

Induction of cyclooxygenase-2 in a rat gastric epithelial cell line by epiregulin and basic fibroblast growth factor.

Prostaglandins play an important role in maintaining gastric mucosal integrity. Cyclooxygenases (COX-1 and -2) are the key enzymes involved in prostaglandin synthesis. COX-2 expression in gastric epithelial cells remains a subject of controversy, and a possible regulation of gastric COX-2 by growth factors has not been explored. Therefore, we studied the effect of growth factors including epiregulin, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) on expression of COX-2 in a gastric epithelial cell line (RGM1) derived from normal rat gastric mucosa. Cells were incubated with 10 or 100 ng/ml of EGF. epiregulin, bFGF, or VEGF for 1, 2, 3, 6, and 24 h. COX-2 mRNA expression was determined by RT-PCR using specific COX-2 primers and COX-2 protein expression was determined by Western blotting. This study showed that COX-2 mRNA and protein are expressed in the gastric epithelial RGM1 cell line and that epiregulin and bFGF (but not VEGF) significantly increase expression of COX-2 mRNA and protein. Because PGs play an important role in mucosal defense, this study suggests that some growth factors contribute to maintaining mucosal integrity via activation of the COX-2 gene.

Animals↗

Relationship between glutathione and polyamine metabolism after partial hepatectomy in alcohol-treated rats.

In a liver regeneration inhibiting model after partial hepatectomy in rats, we determined hepatic glutathione concentration and studied its relation with polyamine, an indicator of liver regeneration. Experiment 1: Male ODS rats and Wistar rats as control were maintained with vitamin C-deficient diets for 2 weeks. In the alcohol group, 3 g/kg of ethanol was administered orally 1 hr before partial hepatectomy. Experiment 2: Wistar rats were divided into four groups, according to the presence or absence of alcohol and vitamin E (VE), and a 6-week pair-feeding was done. As for partial hepatectomy, about 70% of the total liver was excised. As a result, in experiment 1, hepatic glutathione levels were significantly decreased by acute alcohol administration in both ODS and Wistar rats, and its level of alcohol administration group of ODS rats was the lowest. Hepatic ornithine decarboxylase activity and putrescine level 4 hr after partial hepatectomy showed almost a similar behavior, being the lowest in the alcohol administration group of ODS rats. In experiment 2, hepatic glutathione level showed no influence by chronic alcohol administration, but was the lowest in the VE-deficient alcohol administration group. Hepatic ODC activity and putrescine level 4 hr after partial hepatectomy showed no influence by chronic alcohol administration, but was the lowest in the VE-deficient alcohol administration group. These results clarified that intrahepatic glutathione levels before partial hepatectomy were decreased in the models in which hepatic ODC activities after surgery were suppressed.

Animals↗

Rapid serodiagnosis of Mycobacterium avium-intracellulare complex infection by ELISA with cord factor (trehalose 6, 6'-dimycolate), and serotyping using the glycopeptidolipid antigen.

Mycobacterium avium-intracellulare complex (MAC) is one of the most important opportunistic pathogens, particularly in patients with acquired immunodeficiency syndrome (AIDS). The aim of this study was to determine whether an enzyme-linked immunosorbent assay (ELISA) using trehalose 6,6'-dimycolate (TDM) as an antigen can be used for the rapid serodiagnosis of MAC infection. We also identified MAC serotypes by ELISA using serotype-specific glycopeptidolipid (GPL) antigen. To confirm our findings, the thin-layer chromatographic (TLC) behavior of serotype-specific GPL of the strains isolated from MAC-infected patients was also tested. Forty patients infected with MAC and 30 healthy controls were tested. Thirty-two of the 40 MAC-infected patients had higher titers of serum antibodies against MAC TDM than against MTB TDM, while all 30 healthy control sera were unreactive to MAC TDM and MTB TDM. Results of the GPL ELISA indicated that 20 of the 40 MAC-infected patients' sera were reactive against serotype 4 GPL, 3 against serotype 8 GPL, and 1 against serotype 16 GPL. A TLC analysis of the GPL of the 40 MAC isolates showed that 16 strains were of serotype 4, 5 of serotype 8, and 2 of serotype 16. Results of the GPL ELISA were in good accord with those of the TLC analysis for most patients. Our findings suggest that ELISA using TDM is useful for rapid serodiagnosis of MAC infection, and that complementary ELISA testing using serotype-specific GPL gives additional detailed information concerning MAC serotypes.

Adult↗

Phorbol ester-induced G1 arrest in BALB/MK-2 mouse keratinocytes is mediated by delta and eta isoforms of protein kinase C.

We investigated the possible negative regulation of the cell cycle by protein kinase C (PKC) isoforms in synchronously grown BALB/MK-2 mouse keratinocytes, in which PKC isoforms were overexpressed by using the adenovirus vector Ax. Cells at the G1/S boundary of the cell cycle were the most sensitive to the inhibitory effect of 12-O-tetradecanoylphorbol-13-acetate (TPA), a PKC agonist, resulting in G1 arrest. TPA-induced inhibition of DNA synthesis was augmented by overexpression of the eta and delta isoforms, but rescued by the dominant-negative and antisense eta isoforms. In contrast, the alpha and zeta isoforms showed no effect on DNA synthesis with or without TPA treatment. Immunoblotting indicated cell cycle-dependent expression of the eta isoform, being highest in cells at the G1/S boundary. The present study provides evidence that the eta and delta isoforms of PKC are involved in negative regulation of cell cycle at the G1/S boundary in mouse keratinocytes.

Adenoviridae↗

Clinicopathological criteria for multicentricity of hepatocellular carcinoma and risk factors for such carcinogenesis.

Multicentric occurrence is an important characteristic of hepatocellular carcinoma. We evaluated clinicopathological criteria for multicentric hepatocellular carcinoma and identified risk factors for such carcinogenesis. Subjects were 251 consecutive patients undergoing liver resection for hepatocellular carcinoma. One kind of multicentric hepatocellular carcinoma had at least one tumor consisting of well-differentiated hepatocellular carcinoma, together with moderately or poorly differentiated hepatocellular carcinoma located in a separate region. The other kind had an area of well-differentiated component around hepatocellular carcinoma with less differentiation in all occurrences. The outcome of patients with tumors classified in this way was studied. Univariate and multivariate analyses were done to identify risk factors for multicentric hepatocellular carcinoma. The cumulative survival rate was significantly higher in patients with multicentric hepatocellular carcinoma than in patients with hepatocellular carcinoma associated with intrahepatic metastasis. Analysis by Cox's proportional hazard model showed that multicentricity was not a factor in the outcome. The risk of multicentric occurrence increases with progression of chronic liver disease. Univariate analysis showed hepatitis C virus marker and hepatitis B core antibody to be risk factors. By multivariate analysis, the odds ratio for multicentric occurrence in patients infected with hepatitis C virus and with serum hepatitis B virus core antibody compared with patients without either hepatitis C virus or hepatitis B virus was 10.86. This ratio in patients with hepatitis C virus alone was 4.30. These criteria for multicentric hepatocellular carcinoma seem to be clinically useful. Hepatitis C virus infection with or without former infection by hepatitis B virus is a strong risk factor for multicentric hepatocarcinogenesis.

Aged↗

Ulcerative colitis with skip lesions at the mouth of the appendix: a clinical study.

OBJECTIVE: We examined the clinical characteristics of ulcerative colitis patients who demonstrated endoscopically discontinuous lesions at the mouth of the appendix. METHODS: Of patients with initial or recurrent active ulcerative colitis who underwent total colonoscopy during the past 3 yr at Osaka City General Hospital, we selected those who had skip lesions in the mouth of the appendix before treatment, and examined their gender, age, disease type, sites of lesions, inflammatory reaction, severity of disease, effects of treatment, and posttreatment course. RESULTS: Discontinuous lesions at the mouth of the appendix were found in 10 patients, who had the following common clinical features: the major lesion was usually present in the lower part of the large bowel including the rectum, many of the patients had suffered an initial attack only, all patients had mild disease, and many of the patients responded quite satisfactorily to treatment with salicylazosulfapyridine. CONCLUSION: Numerous patients with ulcerative colitis with discontinuous lesions at the mouth of the appendix were observed and their clinical characteristics were examined. Determination of the clinical significance of skip lesions in the appendix will contribute to elucidation of the pathogenesis of ulcerative colitis.

Adult↗

[Isolation of Legionella and free-living amoebae at hot spring spas in Kanagawa, Japan].

Microbiological contamination of hot spring bath water is a public health concern. A province-wide survey was carried out to determine the extent and distribution of both Legionella and free-living amoebae contamination. Among 30 samples of hot spring bath from 12 sites in Kanagawa, Japan, L. pneumophila was detected in 21 water samples from 11 sites, ranging from 10(1)-10(3) CFU/100 ml. Serogroups 3, 5 and 6 of L. pneumophila were predominantly isolated from the samples. Naegleria (46.7%), Platyamoeba (33.3%), Acanthamoeba (10.0%) and 2 other genera of free-living amoebae were detected in 22 samples from 11 sites. One or more genera of host amoebae of Legionella occurred in 17 samples (56.7%) from 9 sites. Another thing to be noted is that 13 water samples contained N. lovaniensis. Although N. lovaniensis is nonpathogenic, it is considered an indicator organism for places that are suitable for the growth of N. fowleri, a causative agent of primary amoebic meningoencephalitis in man.

Amoeba↗

[Occurrence of free-living amoebae and Legionella in whirlpool bathes].

Occurrence of both Legionella species and free-living amoebae were surveyed in whirlpool bathes installed in 11 private houses, 8 public bathes and 13 spas. Free-living amoebae that are known to be the hosts of Legionella were isolated from 24 out of 32 water samples (75%). Single Legionella species, L. pneumophila, with different serogroups (SG) predominantly SG3 (18.3%), SG5 (23.7%) and SG6 (15.8%), were isolated from 21 damples, ranging from 10(1) to 10(4) CFU/100 ml. Further studies were conducted for 10 consecutive weeks to monitor the occurrence of both free-living amoebae and Legionella in the whirlpool bathes of 4 private houses. Free-living amoebae, such as Hartmannella and Vexillifera, and L. pneumophila SG1, SG3, SG4, SG5 and SG6 were consistently isolated from all the water samples throughout the monitoring periods. Bath basins in which Hartmennella and Vannella were isolated tended to harbor large number of Legionella. Management practices such as frequent washing filter elements and/or frequent addition of tap water to bath basins is highly recommended to reduce microbial contaminants.

Amoeba↗

[Experimental infection of Naegleria fowleri in mice].

Ten SPF mice (ddY, 4w-old, female) were infected by nasal instillation with an isolate of Naegleria fowleri that was first isolated from a patient with primary amoebic meningoencephalitis (PAM) in Japan. Of these mice, 2 showed clinical signs typical for PAM on the 4th day. On the next day, 5 mice became very ill and remained immobile; their movement and response to painful stimuli diminished progressively. All the infected mice were then examined histopathologically on the same day regardless of their clinical signs. Pathological changes due to invasion and/or proliferation of amoebae were observed in 5 mice with clinical signs. Swelling of the nasal mucosa and ulcerated nasal epithelium with inflammatory cells were observed. Proliferation of amoebae was detected to a lesser extent in nasal cavity including mucous membrane and nasal epithelium. Olfactory lobes and arteriolar hemisphere were necrotic with haemorrhage and filled with amoebae. From these findings the pathogenicity of the isolate was confirmed to develop PAM in experimental animals. It was also observed that the olfactory neuroepithelium was the route of invasion in PAM due to N. fowleri and consequently migration occurred through olfactory lobes into the cerebrum.

Amebiasis↗

Increased production of interleukin-10 by human blood monocytes stimulated with Mycobacterium avium-intracellulare complex.

Macrophages produce various cytokines in response to mycobacteria, including interleukin 10 (IL-10) and tumor necrosis factor alpha (TNF-alpha). IL-10 has been shown to down-regulate numerous macrophage functions, including microbicidal activity against intracellular bacteria and parasites. IL-10 also inhibits interferon-gamma (IFN-gamma) production and antigen-specific proliferation of Th1 cells mediating immunologic resistance to mycobacterial infection. In contrast, TNF-alpha activates macrophages and may augment their mycobacterial activity. In this study, peripheral blood mononuclear cells (PBMC) or blood monocytes obtained from healthy tuberculin reactors were stimulated in vitro with heat-killed Mycobacterium tuberculosis or heat-killed M. avium-intracellulare complex (MAC) to produce IL-10 and TNF-alpha. We studied a total of 26 clinical isolates of M. tuberculosis and 28 isolates of MAC. MAC-stimulated PBMC and monocytes released significantly larger amounts of IL-10 than those cells stimulated with M. tuberculosis. However, there was no difference in induction of TNF-alpha production between MAC and M. tuberculosis. When TNF-activity was neutralized by the addition of anti-TNF-alpha mAb in culture, MAC still induced more IL-10 secretion than did M. tuberculosis. These findings suggest that increased production of IL-10 by MAC-stimulated monocytes may play a role in the intractable disease caused by these organisms.

Cells, Cultured↗

Induction of differentiation in normal human keratinocytes by adenovirus-mediated introduction of the eta and delta isoforms of protein kinase C.

Protein kinase C (PKC) plays a crucial role(s) in regulation of growth and differentiation of cells. In the present study, we examined possible roles of the alpha, delta, eta, and zeta isoforms of PKC in squamous differentiation by overexpressing these genes in normal human keratinocytes. Because of the difficulty of introducing foreign genes into keratinocytes, we used an adenovirus vector system, Ax, which allows expression of these genes at a high level in almost all the cells infected for at least 72 h. Increased kinase activity was demonstrated in the cells overexpressing the alpha, delta, and eta isoforms. Overexpression of the eta isoform inhibited the growth of keratinocytes of humans and mice in a dose (multiplicity of infection [MOI])-dependent manner, leading to G1 arrest. The eta-overexpressing cells became enlarged and flattened, showing squamous cell phenotypes. Expression and activity of transglutaminase 1, a key enzyme of squamous cell differentiation, were induced in the eta-overexpressing cells in dose (MOI)- and time-dependent manners. The inhibition of growth and the induction of transglutaminase 1 activity were found only in the cells that express the eta isoform endogenously, i.e., in human and mouse keratinocytes but not in human and mouse fibroblasts or COS1 cells. A dominant-negative eta isoform counteracted the induction of transglutaminase 1 by differentiation inducers such as a phorbol ester, 1alpha,25-dihydroxyvitamin D3, and a high concentration of Ca2+. Among the isoforms examined, the delta isoform also inhibited the growth of keratinocytes and induced transglutaminase 1, but the alpha and zeta isoforms did not. These findings indicate that the eta and delta isoforms of PKC are involved crucially in squamous cell differentiation.

Adenoviruses, Human↗

Nitric oxide regulates energy metabolism and Bcl-2 expression in intestinal epithelial cells.

Nitric oxide (NO) inhibits the respiration of mitochondria and enteric bacteria, particularly under low O2 concentration, and induces apoptosis of various types of cells. To gain insight into the molecular role of NO in the intestine, we examined its effects on the respiration, Ca2+ status, and expression of Bcl-2 in cultured intestinal epithelial cells (IEC-6). NO reversibly inhibited the respiration of IEC-6 cells, especially under physiologically low O2 concentration. Although NO elevated cytosolic Ca2+ as determined by the fura 2 method, the cells were fairly resistant to NO. Kinetic analysis revealed that prolonged exposure to NO elevated the levels of Bcl-2 and suppressed the NO-induced changes in Ca2+ status of the cells. Because Bcl-2 possesses antiapoptotic function, toxic NO effects might appear minimally in enterocytes enriched with Bcl-2. Thus NO might effectively exhibit its antibacterial action in anaerobic intestinal lumen without inducing apoptosis of Bcl-2-enriched mucosal cells.

Animals↗