Biomedical subjects

Takashi Hara

Publications and source records attributed to Takashi Hara.

At least 19 recordsLinked to original sources

PubMed2006

Structure and expression of two genes encoding secreted acid phosphatases under phosphate-deficient conditions in Pholiota nameko strain N2.

Twenty-three polypeptides secreted in response to a deficiency of inorganic phosphate (Pi) were previously found by two-dimensional polyacrylamide gel electrophoresis analysis in mycelia of Pholiota nameko strain N2. In this study, N-terminal sequencing revealed three of them to be identical to known acid phosphatases of P. nameko strain N114. Two cDNAs and the corresponding genomic DNAs of genes PNAP1 and PNAP2 which encode two of the three acid phosphatases were cloned. The deduced amino acid sequences of PNAP1 and PNAP2 showed high similarity to other fungal acid phosphatases and contained a putative catalytic active site of acid phosphatase. PNAP1 and PNAP2 are comprised of five and seven exons interrupted by four and six introns, respectively. Their promoter regions include two cis-acting elements found in Pi deficiency-inducible genes of Saccharomyces cerevisiae, together with several known functional elements such as a TATA box. Northern blot analysis showed that PNAP1 and PNAP2 are expressed in response to a deficiency of Pi.

Acid Phosphatase↗

PubMed2005

A modified stapling technique for performing Billroth I anastomosis after distal gastrectomy.

Autosuture staplers have provided various operative and postoperative benefits in gastrointestinal surgery. We describe herein our technique of performing distal gastrectomy followed by side-to-end gastroduodenostomy. Eighteen consecutive patients with gastric cancer underwent distal gastrectomy by the staple technique. We propose a modification of the stapling technique to reduce the number of devices used. In our technique, side-to-end anastomosis is performed first, followed by gastric resection just distal to the anastomosis. By introducing the stapler through the area to be resected, we can eliminate the need to close the gastrostomy. Anastomotic stricture occurred in only 1 (5.5%) of 18 cases. There have been no other postoperative complications related to the anastomosis. We believe that our technique is superior in simplicity and security to the conventional techniques.

Gastrectomy↗

PubMed2004

Determination of HIV type 1 CRF01_AE gag p17 and env-V3 consensus sequences for HIV/AIDS vaccine design.

A molecular epidemiological study of the gag p17 and env-V3 regions on HIV-infected drug users and blood donors was carried out in northern Thailand from 1998 through 2002 to determine the predominant subtype and consensus sequence (CS) for circulating HIV-1 strains. CRF01_AE was concluded to be a predominant strain and the nucleotide CSs in gag p17 and env-V3 showed only 1.26% and no difference from CS in the Los Alamos database, respectively. Our env-V3 CS was identical to the previously published CSs, suggesting that the CS was very conserved from 1990 through 2002 in Thailand. Gag p17 and env-V3 nucleotide sequences of seroconvertors in our subjects were quite similar to the CS and conserved for at least 9 and 6 years postinfection, respectively. These results suggest that the CS approach to the HIV-1 antigen design could overcome HIV diversity and help us develop an effective HIV/AIDS vaccine.

AIDS Vaccines↗

PubMed2004

Clinical studies of strangulating small bowel obstruction.

Reliable preoperative diagnosis of intestinal necrosis in strangulating small bowel obstruction (SSBO) is difficult, and, as yet, no reliable marker has been described. We, therefore, retrospectively examined clinical symptoms and hematobiochemical data of patients with SSBO in our surgical wards. Thirty-seven patients with SSBO were analyzed in this study. They were divided into two groups: group A (13 patients), the presence of gangrenous intestine; and group B (24 patients), the absence of it. By means of chi2 test, Student t test, or Welch t test, peritoneal signs, white blood cell count (leukocytosis or leukopenia), systemic inflammatory response syndrome (SIRS), shock, and base deficit were significantly associated with whether gangrenous intestine existed or not. Next, in simple regression analysis, base deficit was significantly correlated with the length of gangrenous intestine. In stepwise logistic regression analysis, SIRS was independently correlated with the presence of gangrenous intestine. If SIRS or metabolic acidosis is seen in patients with SSBO, the intestine is certainly gangrenous.

Acidosis↗

PubMed2004

[Preliminary reports on psychiatric practice in correctional facilities].

In October 2001, Nanashakon, a council composed of seven psychiatry-related organizations in Japan, decided to launch an investigation into forensic psychiatry in Japan, and established a working team (WT) for this purpose. From its establishment to March 2004, the WT performed surveys and analyses of the current situation of preliminary reports by psychiatric experts (preliminary reports) and of psychiatric practice in correctional facilities. Based on the results, the WT has presented proposals including guidelines for preliminary reports. In January 2002, the WT conducted an awareness survey on the status quo of testimony by psychiatric experts and forensic psychiatry, targeting the members of the Japanese Society of Psychiatry and Neurology, and obtained 666 replies. The survey revealed various critical opinions such as skepticism over the current punishment imposed on criminal patients with mental disorders. In February 2002, the WT obtained data on preliminary reports (2,042 cases) compiled prior to prosecution in FY2000 from the Japanese Ministry of Justice. Reviewing the details and differences between the evaluation by psychiatrists and the decision by public prosecutors, the WT pointed out the ambiguity of criteria used for the evaluation of competency of weak-minded persons and the criteria for criminal punishment. Around the same time, the WT was also asked by a news agency to analyze the preliminary reports of 50 district public prosecutor offices. The results revealed marked regional differences in the operation of the preliminary evaluation system for competency. In January 2003, the WT collected 146 preliminary reports from around the country for comparison and review, and again found conspicuous individual and regional discrepancies in the format and content. Based on these results, the WT conducted a hearing of 41 expert opinions on preliminary reports, and in January 2004, proposed guidelines outlining a format model of preliminary reports, and a training and authorizing system for forensic psychiatrists, to standardize preliminary reports and enhance their reliability. In February 2004, the WT conducted a questionnaire survey on the current situation of psychiatry in correctional facilities, targeting doctors with experience working under these circumstances. Fifty-one replies were obtained. Most of the respondents approved of the current system. However, to incite arguments in this area, attempts were made to draw critical responses and discussion by presenting data on the current situation of psychiatric practice in correctional facilities.

Criminal Law↗

PubMed2003

Structure and expression of a phosphate deficiency-inducible ribonuclease gene in Pholiota nameko.

Thirty-one cDNAs corresponding to pdi genes [inorganic phosphate (Pi) deficiency-inducible genes] were previously isolated through the differential screening of a cDNA library constructed from the mycelium of Pholiota nameko. Among the cDNAs, pdi370 was analyzed here. The deduced amino acid sequence showed high similarity to fungal ribonucleases (RNases) and contained two signature sequences conserved in T2 family RNases: CAS1 and CAS2. Genomic DNA harboring the pdi370 gene was isolated from a genomic library of P. nameko. Sequence analysis showed that the pdi370 gene is interrupted by 16 introns and that the promoter region contains two cis-acting sequences found in Pi deficiency-induced genes from Saccharomyces cerevisiae, together with several known functional elements, such as a TATA box. RNase activity in the mycelium and culture filtrate increased 5.6-fold and 5.2-fold, respectively, under Pi-deficient conditions. Staining for RNase activity showed that at least four RNases are induced and secreted under the conditions. The N-terminal sequence of one of them agreed with that of the pdi370 gene product.

Agaricales↗

PubMed2003

Three genes specifically expressed during phosphate deficiency in Pholiota nameko strain N2 encode hydrophobins.

We previously isolated 31 cDNAs corresponding to pdi [inorganic phosphate (Pi) deficiency- inducible] genes through the differential screening of a cDNA library constructed from the mycelium of Pholiota nameko strain N2 cultured in Pi-depleted medium. Among the cDNAs, pdi251, pdi263 and pdi315 were analyzed here. The deduced amino acid sequences of pdi251, pdi263 and pdi315 showed high similarity to fungal hydrophobins and genes corresponding to these cDNAs encoding P. nameko hydrophobins were designated pnh1, pnh2 and pnh3, respectively. PNH1, PNH2 and PNH3 had a conserved spacing of eight cysteine residues in hydrophobins and a hydropathy pattern characteristic of class I hydrophobins. Phylogenetic analysis showed that PNH1, PNH2 and PNH3 were phylogenetically similar and significantly related to the hydrophobin POH1 specifically expressed in fruiting bodies of Pleurotus ostreatus. Northern blot analysis indicated that, under conditions of Pi deficiency, pnh2 and pnh3 were also induced in strains N4 and N301.

Agaricales↗

PubMed2003

Mechanism underlying the inner membrane retention of Escherichia coli lipoproteins caused by Lol avoidance signals.

Escherichia coli lipoproteins are localized to either the inner or outer membrane depending on the residue at position 2. The inner membrane retention signal, Asp at position 2 in combination with certain residues at position 3, functions as a Lol avoidance signal, i.e. the signal inhibits the recognition of lipoproteins by LolCDE that releases lipoproteins from the inner membrane. To understand the role of the residue at position 2, outer membrane-specific lipoproteins with Cys at position 2 were subjected to chemical modification followed by the release reaction in reconstituted proteoliposomes. Sulfhydryl-specific introduction of nonprotein molecules or a negative charge to Cys did not inhibit the LolCDE-dependent release. In contrast, oxidation of Cys to cysteic acid resulted in generation of the Lol avoidance signal, indicating that the Lol avoidance signal requires a critical length of negative charge at the second residue. Furthermore, not only modification of the carboxylic acid of Asp at position 2 but also that of the amine of phosphatidylethanolamine abolished the Lol avoidance function. Based on these results, the Lol avoidance mechanism is discussed.

Amino Acid Sequence↗

PubMed2003

Malignant mixed Mullerian tumor of the ovary growing into an inguinal hernia sac: report of a case.

Malignant tumors presenting as an inguinal hernia are rare. We present the case of a malignant mixed Mullerian tumor (MMMT) of the ovary growing into an inguinal hernia sac. In this case, magnetic resonance imaging was useful in making a diagnosis of an ovarian neoplasm growing into the inguinal canal, and to the best of our knowledge, this is only the tenth case of a malignant ovarian tumor and the first case reported in the English-language literature of MMMT of an ovary which grew into an inguinal hernia sac.

Aged↗

PubMed2003

Usefulness of flexible colonoscopic microwave coagulation therapy for a colorectal anastomotic stricture.

PURPOSE: Several methods of treatment for benign anastomotic strictures after anterior resection have been described. We describe a simple, safe, effective, and inexpensive method for treating benign colorectal anastomotic stricture by means of microwave coagulation under flexible colonoscopic visualization. METHODS: Eighteen patients with rectal or rectosigmoidal cancer underwent low anterior resection or anterior resection without colonic pouch and colorectal anastomosis by a double-stapling technique with PCEEA. Two of 18 patients (11.1 percent) developed an anastomotic stenosis. A microwave electrode was passed through the biopsy channel of the flexible colonoscope. Under flexible colonoscopic visualization, microwave irradiation was performed at four points (3, 6, 9, and 12 o'clock) in the stricture site because of granulation scar to obtain an adequately coagulated area. No bougies were performed thereafter. RESULTS: The anastomotic strictures could be dilated adequately, the patients could defecate satisfactorily, and their abdominal distentions and bowel symptoms were resolved. No complications occurred. No occurrence of restricture has been observed. CONCLUSION: Flexible endoscopic, microwave coagulation therapy is a useful, simple, effective, and safe method for the treatment of benign colorectal anastomotic strictures.

Anastomosis, Surgical↗

PubMed2003

Assessment of oral transmission using cell-free human immunodeficiency virus-1 in mice reconstituted with human peripheral blood leucocyte.

Oral-genital contact is one of the risk factors for the transmission of human immunodeficiency virus (HIV) in adults. In recent reports, oral exposure to simian immunodeficiency virus (SIV) was found to have important implications for the achievement of mucosal transmission of HIV in a rhesus monkey animal model. In the present study, we aimed first to establish a small animal model which did not develop tonsils suitable for HIV oral mucosa transmission, using non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice and NOD/SCID B2m(null) mice grafted with human peripheral blood leucocytes (hu-PBL) and stimulated with interleukin (IL)-4, and second to investigate whether oral exposure to cell-free R5 and X4 HIV-1 could lead to oral transmission of HIV through intact or traumatized mucosal tissues in humanized mice. Both low and high concentrations of cell-free R5 and X4 viruses failed to cause oral transmission with or without trauma in hu-PBL-NOD/SCID and NOD/SCID Beta2m(null) mice, which presented a number of CD4+ cells in gingival tissues and oral cavities with or without tissue injury. The present results show that IL-4-administrated NOD/SCID B2m(null) mice are useful as a small-humanized model for the study of HIV oral infection, which may help to define the window of opportunity for oral transmission by the HIV virus in animal model experiments.

Animals↗

PubMed2003

Distal pancreatectomy: is staple closure beneficial?

BACKGROUND: Appropriate closure of the pancreatic remnant after distal pancreatectomy remains controversial. Data on distal pancreatectomy were reviewed to evaluate the effectiveness of staple closure compared with suture closure. METHODS: Thirty-three patients underwent distal pancreatectomy between January 1994 and December 2001. They were subdivided according to the method of closure of the distal pancreatic stump: the staple group comprised 10 patients and the suture group comprised 23 patients. Charts were reviewed for the method of closure of the distal stump as well as for the serum and urinary amylase level, mortality and morbidity. RESULTS: In the staple group, no patient developed a pancreatic fistula, whereas in the suture group, eight patients (33.3%) developed a pancreatic fistula (P = 0.0353). The serum amylase level (mean +/- SD) on the first postoperative day was 185 +/- 71 IU/L in the staple group and 499 +/- 461 IU/L in the suture group (P = 0.0413). CONCLUSION: Staple closure by means of a Powered Multifire Endo GIA 60 is a simple, quick and safe alternative to the standard suture closure technique, as it reduces the incidence of pancreatic fistula.

Adolescent↗

PubMed2003

The diagnosis and treatment of adult intussusception.

BACKGROUND: Intussusception in adults is often diagnosed on computed tomography (CT), and the optimal treatment of this entity is not universally agreed upon. We report our experience in an attempt to clarify the usefulness of CT scan and the optimal treatment of this entity. STUDY: Seven cases of adult intussusception were encountered at our institute between 1991 and 2001. Data related to presentation, diagnosis, treatment, and pathology were analyzed. RESULTS: Preoperative diagnosis was made in four patients by CT scan and/or ultrasonography. Two patients had colonic cancer and one had jejunal cancer. Three of four patients with small bowel intussusception underwent reduction before resection and the other one underwent resection without reduction because of severe ischemic bowel. CONCLUSIONS: The CT scan is most useful in making the diagnosis of intussusception. Colonic lesions should be resected without reduction. Small bowel lesions should be reduced only in patients in whom a benign diagnosis has been strongly suggested preoperatively or in patients in whom resection may result in short gut syndrome.

Adenocarcinoma↗

PubMed2003

Helicobacter pylori infection and early gastric cancer.

BACKGROUND: Helicobacter pylori has recently been associated with an increased risk of gastric cancer. This study aimed to examine the association between H. pylori, histologic chronic gastritis, and intestinal metaplasia in early gastric cancers of different histologic types. STUDY: Seventy-four patients who were surgically diagnosed as having early gastric cancer were included in this study. All tissue specimens were obtained from patients by endoscopic biopsy and were classified histopathologically as intestinal-type early gastric cancer in 55 patients and diffuse-type early gastric cancer in 19 patients. RESULTS: H. pylori infection was found in 67 patients (90.5%) but not found in seven (9.5%). And the prevalence of H. pylori infection with nongastric cancer patients was also high (68.5%). There was no significant difference between the intestinal-type and the diffuse-type early gastric cancer in chronic active gastritis and atrophic chronic gastritis. Intestinal metaplasia was observed more frequently in patients with the intestinal-type than with the diffuse-type early gastric cancer (P = 0.0102). CONCLUSIONS: Infection with H. pylori has an important relationship to both histopathologic types of early gastric cancer.

Adult↗

PubMed2003

[Flashback].

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Amphetamine-Related Disorders↗

PubMed2002

First report of CRF03_AB recombinant HIV type 1 in injecting drug users in Ukraine.

It has been reported that subtypes A throughout H of HIV-1 are circulating in the former Soviet Union. In this sequence note, we analyzed the genetic prevalence of HIV-1 among injecting drug users (IDUs) in Ukraine. The subjects studied included two individuals from Kiev and six individuals from Simferopol', the latter located in the Crimean Peninsula. We found that one of eight IDUs was infected with a CRF03 gagA/envB recombinant HIV-1 and was from Simferopol', whereas the others were infected with HIV-1 subtype A. There combinant was closely related to other A/B recombinants reported previously, and had silent mutations Inthe V3 region, the same as other envB strains of HIV-1 circulating among mDUs in the former Soviet Union. The data supported reports that the Russian AIB recombinant HIV-1 was probably from Ukraine. This is the first report of a CRF03 gagA/envB recombinant HIV-1 found in Ukraine.

Amino Acid Sequence↗

PubMed2002

Aminoacylation of the N-terminal cysteine is essential for Lol-dependent release of lipoproteins from membranes but does not depend on lipoprotein sorting signals.

Lipoproteins are present in a wide variety of bacteria and are anchored to membranes through lipids attached to the N-terminal cysteine. The Lol system of Escherichia coli mediates the membrane-specific localization of lipoproteins. Aspartate at position 2 functions as a Lol avoidance signal and causes the retention of lipoproteins in the inner membrane, whereas lipoproteins having residues other than aspartate at position 2 are released from the inner membrane and localized to the outer membrane by the Lol system. Phospholipid:apolipoprotein transacylase, Lnt, catalyzes the last step of lipoprotein modification, converting apolipoprotein into mature lipoprotein. To reveal the importance of this aminoacylation for the Lol-dependent membrane localization, apolipoproteins were prepared by inhibiting lipoprotein maturation. Lnt was also purified and used to convert apolipoprotein into mature lipoprotein in vitro. The release of these lipoproteins was examined in proteoliposomes. We show here that the aminoacylation is essential for the Lol-dependent release of lipoproteins from membranes. Furthermore, lipoproteins with aspartate at position 2 were found to be aminoacylated both in vivo and in vitro, indicating that the lipoprotein-sorting signal does not affect lipid modification.

ATP-Binding Cassette Transporters↗

PubMed2002

Purification and characterization of lipoxygenase from Pleurotus ostreatus.

Lipoxygenase was purified homogeneously from cups of Pleurotus ostreatus by Sephacryl S-400 HR gel filtration, Dyematrex Green A affinity, and DEAE-Toyopearl 650M ion-exchange chromatographies. The molecular weight of the enzyme was estimated to be 67,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 66,000 by gel filtration; the isoelectric point was pH 5.1. The optimum pH and temperature of the enzymatic activity were 8.0 and 25 degrees C, respectively. The enzyme contained non-heme iron, and a thiol group seemed to be involved in its activity. The K(m), V(max), and k(cat) values of the enzyme for linoleic acid were 0.13 mM, 23.4 micromol.min(-1).mg(-1), and 25.7 s(-1), respectively. The enzyme showed high specificity toward linoleic acid. When linoleic acid was incubated with the enzyme, 13-hydroperoxy-9Z,11E-octadecadienoic acid was found to be the main oxidative product.

Chromatography, Affinity↗