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Biomedical subjects

M Kobayashi

Publications and source records attributed to M Kobayashi.

At least 199 records · Page 11Linked to original sources
PubMed2001

Mycobacterium bovis Bacillus Calmette-Guerin and its cell wall complex induce a novel lysosomal membrane protein, SIMPLE, that bridges the missing link between lipopolysaccharide and p53-inducible gene, LITAF(PIG7), and estrogen-inducible gene, EET-1.

LITAF and PIG7 encode an identical protein, and they have recently been reported as lipopolysaccharide and p53-inducible genes, respectively. By using the differential display approach, we identified a Mycobacterium bovis BCG cell wall skeleton (BCG-CWS)-inducible gene fragment from human monocytes, showing no homology to any reported gene. Full-length cloning of this fragment reveals the following. 1) The differential display product represents the incomplete 3'-untranslated region of LITAF/PIG7. 2) The coding region of the transcript differs from LITAF/PIG7 due to an absence of a single guanine residue, resulting in a potential translational frameshift. 3) The newly coded protein turns out to be 86% identical and 90% similar to an estrogen-inducible rat gene, EET-1. Repeated analysis, expressed sequence tag search, comparison with homologues, and genome sequence analysis confirmed the absence of the single guanine residue. One interesting feature of this protein is that it possesses the RING domain signature and is predicted to be localized in the nucleus. However, detailed analysis together with experimental evidence suggests it is neither a RING family member nor a nuclear protein. Comparison of a total collection of 18 proteins from various species indicates that proteins of this family are small in size and mainly conserved at the C-terminal domain with a unique motif. We characterize this novel protein as an unglycosylated small integral membrane protein of the lysosome/late endosome (SIMPLE) whose expression is elicited in monocytes by live and heat-killed BCG, BCG cell wall complex, lipopolysaccharide, and tumor necrosis factor-alpha. To our knowledge this is the first report of pathogen-associated molecular pattern (PAMP)-induced differential expression of a lysosomal membrane protein presumably involved in apoptosis.

Amino Acid Sequence↗
PubMed2001

Correlation between loose density and compactibility of granules prepared by various granulation methods.

The objectives of this study were to prepare the lactose granules by various granulation methods using polyethylene glycol 6000 (PEG 6000) as a binder and to evaluate the effects of granulation methods on the compressibility and compactibility of granules in tabletting. Lactose was granulated by seven granulation methods -- four wet granulations including wet massing granulation, wet high-speed mixer granulation, wet fluidized bed granulation and wet tumbling fluidized bed granulation; and three melt granulations including melt high-speed mixer granulation, melt fluidized bed granulation and melt tumbling fluidized bed granulation. The loose density, angle of repose, granule size distribution, mean diameter of granules, and the tensile strength and porosity of tablets were evaluated. The compactibilities of granules were varied by the granulation methods. However, the difference in compactibility of granules could not be explained due to the difference in compressibility, since there was no difference in Heckel plots due to granulation methods. Among their granule properties, the loose density of granules seemed to have a correlation with the tablet strength regardless of the granulation methods.

Chemistry, Pharmaceutical↗
PubMed2001

Effect of magnesium stearate or calcium stearate as additives on dissolution profiles of diltiazem hydrochloride from press-coated tablets with hydroxypropylmethylcellulose acetate succinate in the outer shell.

Effect of magnesium stearate (MgSt) or calcium stearate (CaSt) on the dissolution profiles of diltiazem hydrochloride in the core of press-coated (PC) tablets with an outer shell composed of hydroxypropylmethylcellulose acetate succinate (HPMCAS) was evaluated by porosity and changes in IR spectra of tablets. In JP first fluid (pH 1.2), the lag time increased with decreasing porosity and was greatest by the addition of MgSt to HPMCAS. While, in JP second fluid (pH 6.8), it increased with decreasing porosity by the addition of CaSt, but hardly changed by the addition of MgSt. Thus, using tablets prepared with the same composition as the outer shell, the changes in IR spectra and uptake amount of the dissolution media after immersion in first fluid and second fluid were determined. The results suggested that some physicochemical interaction occur between MgSt and HPMCAS in tablets with HPMCAS and MgSt and the uptake increased markedly in each dissolution medium. These phenomena seem to cause a prolongation of lag time in first fluid but a shortening of it in second fluid in PC tablets with HPMCAS and MgSt. In contrast, CaSt and HPMCAS did not show such interactions and increased the hydrophobic properties of the outer shell. Consequently, the lag time was only slightly prolonged in first fluid, however, markedly prolonged in second fluid due to suppression of second fluid penetration into micro pores in the outer shell and HPMCAS gel formation on the surface in PC tablets with HPMCAS and CaSt.

Antihypertensive Agents↗
PubMed2001

[Occupationally induced nitric acid and sulfuric acid burns: an analysis of 2 patients from the aspect of occupational health].

We report two patients who suffered from acid burns while working in chemical factories. Case 1: a 44-year-old man who received burn induced by nitric acid on the face and extremities. Despite his protecting facial mask, he was exposed to nitric acid on his face through a gap between the mask and skin surface. Nitric acid was also sprinkled on his scalp which was not covered by a helmet or a protecting device. In addition, he suffered from acid burn on the right scapular region, the right upper arm, and the lower extremities through the work clothes. Case 2: a 26-year-old man who suffered from sulfuric acid burn on the forearms. Both patients were accidentally exposed to acids while they filled tanks with the acids through a hose. Following the manual of the factories, they washed the exposed skin with water for more than 15 minutes after the exposure. Although they recovered without any serious sequel, there remained partial deep tissue destruction of the skin. We reviewed these two cases from the aspect of industrial medicine, and proposed the following three points for improvement in the workplace to prevent accidental acid burns. 1) re-education or enlightenment activities for the well-experienced workers to avoid negligence to the danger of strong acid. 2) recommendation to take a complete shower to avoid overlooking of unaware acid injury. 3) improvement in the protecting facial mask. In addition, clinicians who examine acid-burn patients should not pass over the presence of deep ulcers lying behind the thick crust on the injured area.

Adult↗
PubMed2001

Identification of CRE1 as a cytokinin receptor from Arabidopsis.

Cytokinins are a class of plant hormones that are central to the regulation of cell division and differentiation in plants. It has been proposed that they are detected by a two-component system, because overexpression of the histidine kinase gene CKI1 induces typical cytokinin responses and genes for a set of response regulators of two-component systems can be induced by cytokinins. Two-component systems use a histidine kinase as an environmental sensor and rely on a phosphorelay for signal transduction. They are common in microorganisms, and are also emerging as important signal detection routes in plants. Here we report the identification of a cytokinin receptor. We identified Arabidopsis cre1 (cytokinin response 1) mutants, which exhibited reduced responses to cytokinins. The mutated gene CRE1 encodes a histidine kinase. CRE1 expression conferred a cytokinin-dependent growth phenotype on a yeast mutant that lacked the endogenous histidine kinase SLN1 (ref. 10), providing direct evidence that CRE1 is a cytokinin receptor. We also provide evidence that cytokinins can activate CRE1 to initiate phosphorelay signalling.

Arabidopsis↗
PubMed2001

An in vitro investigation of the suitability of press-coated tablets with hydroxypropylmethylcellulose acetate succinate (HPMCAS) and hydrophobic additives in the outer shell for colon targeting.

To develop a new colon targeting formulation, which can suppress drug release completely during 12 h in the stomach and release the drug rapidly after a lag time of 3+/-1 h in the small intestine, the use of press-coated tablets with hydroxypropylmethylcellulose acetate succinate (HPMCAS) in the outer shell was investigated. The release of diltiazem hydrochloride (DIL) as a model drug contained in the core tablets in the 1st fluid (pH 1.2) was suppressed by preparing with higher compression force, but the lag time in the 2nd fluid (pH 6.8) could not exceed 1.5 h. Therefore, to improve the dissolution characteristics, the effects of addition of various hydrophobic additives to HPMCAS were examined. All of the additives examined suppressed the release rate in the 1st fluid, and prolonged the lag time in the 2nd fluid compared to HPMCAS alone. However, although none of the additives examined fulfilled all of the desired criteria, magnesium stearate (MgSt) and calcium stearate (CaSt) showed interesting effects; the former suppressed drug release completely in 1st fluid, while the latter markedly prolonged the lag time in 2nd fluid. To integrate the merits of each additive, press-coated tablets with a powder mixture of HPMCAS, MgSt and CaSt in the outer shell (HMC tablets) were prepared and in vitro tests were performed. The results indicated that HMC tablets with a mixing ratio of 80% HPMCAS, 5-15% MgSt and 15-5% CaSt in the outer shell met the desired criteria and the lag time in 2nd fluid could also be controlled from 2 to 9 h. At a mixing ratio of 80% HPMCAS, 10% MgSt and 10% CaSt, the dissolution profiles of DIL in 1st fluid and 2nd fluid were not remarkably affected by agitation intensity, and addition of bile salts, pretreatment time or anticipated higher pH except for pH 6.0, respectively. These results indicated the usefulness of HMC tablets with the desirable functions for colon-targeting formulations.

Colon↗
PubMed2001

Oxidation of thymine to 5-formyluracil in DNA promotes misincorporation of dGMP and subsequent elongation of a mismatched primer terminus by DNA polymerase.

5-Formyluracil (fU) is a major oxidative thymine lesion generated by ionizing radiation and reactive oxygen species. In the present study, we have assessed the influence of fU on DNA replication to elucidate its genotoxic potential. Oligonucleotide templates containing fU at defined sites were replicated in vitro by Escherichia coli DNA polymerase I Klenow fragment deficient in 3'-5'-exonuclease. Gel electrophoretic analysis of the reaction products showed that fU constituted very weak replication blocks to DNA synthesis, suggesting a weak to negligible cytotoxic effect of this lesion. However, primer extension assays with a single dNTP revealed that fU directed incorporation of not only correct dAMP but also incorrect dGMP, although much less efficiently. No incorporation of dCMP and dTMP was observed. When fU was substituted for T in templates, the incorporation efficiency of dAMP (f(A) = V(max)/K(m)) decreased to (1/4) to (1/2), depending on the nearest neighbor base pair, and that of dGMP (f(G)) increased 1.1-5.6-fold. Thus, the increase in the replication error frequency (f(G)/f(A) for fU versus T) was 3.1-14.3-fold. The misincorporation rate of dGMP opposite fU (pK(a) = 8.6) but not T (pK(a) = 10.0) increased with pH (7.2-8.6) of the reaction mixture, indicating the participation of the ionized (or enolate) form of fU in the mispairing with G. The resulting mismatched fU:G primer terminus was more efficiently extended than the T:G terminus (8.2-11.3-fold). These results show that when T is oxidized to fU in DNA, fU promotes both misincorporation of dGMP at this site and subsequent elongation of the mismatched primer, hence potentially mutagenic.

Base Pair Mismatch↗
PubMed2001

Efficacy of IFN therapy based on duration period of negative HCV-RNA during IFN administration.

We assessed the relationship between the duration period of negative hepatitis C virus (HCV)-RNA during interferon (IFN) therapy and the efficacy after prolonged IFN therapy in patients with HCV-genotype 1b and high virus load of more than 1 mega equivalents/ml (Meq/ml) retrospectively. A total of 100 patients who had HCV-genotype 1b and a high virus load of more than 1 Meq/ml and were treated with natural IFN-alpha for more than 12 months were enrolled in this trial. These patients were given 6 MU of IFN daily for 8 weeks, followed by three times weekly for another more than 44 weeks. The HCV-RNA pattern during IFN therapy according to negative or positive of the serum HCV-RNA by reverse transcription nested polymerase chain reaction (RT-nested PCR) from 2 months after the initiation of IFN to the termination of IFN were classified as follows: group 1: constant negative HCV-RNA (n=41 cases), group 2: constant positive HCV-RNA (n=35 cases), group 3: HCV-RNA pattern except for group 1 or group 2 (n=24 cases). A complete response (CR) was defined as negative HCV-RNA by RT-nested PCR at two points, 3 and 6 months after the completion of IFN therapy. CR rate was 58.5% (24 cases) in group 1, but CR rate in group 2 or group 3 was 0%. In group 1, the CR rate was 100% (10/10) in patients with negative HCV-RNA constantly for period of more than 24 months during IFN therapy. On the other hand, all patients who had positive HCV-RNA 2 months after the initiation of IFN did not get CR. In conclusion, it seems to us that the attainment of constantly negative HCV-RNA for the period of more than 24 months during IFN therapy is highly related to CR.

Journal Article↗
PubMed2001

Ultrastructure of the interface between bioactive composite and bone: comparison of apatite and wollastonite containing glass-ceramic filler with hydroxyapatite and beta-tricalcium phosphate fillers.

We have developed a bioactive bone cement that consists of apatite and wollastonite containing glass-ceramic (AW-GC) powder and bisphenol-a-glycidyl dimethacrylate (Bis-GMA)-based resin. In this study, we made three types of composite (designated AWC, HAC, and TCPC) consisting of AW-GC, hydroxyapatite (HA,) or beta-tricalcium phosphate (beta-TCP) powder as the inorganic filler and Bis-GMA-based resin as the organic matrix. The proportion by weight of the filler mixed into the cement was 70%. Rectangular plates (10 x 15 x 2 mm) of each composite were made and abraded with 2000 alumina powder. These composites were implanted into tibial metaphyses of rabbits. Specimens were prepared 10 and 25 weeks after implantation and examined using transmission electron microscopy (TEM). AWC was in direct contact with bone 10 weeks after implantation, and AW-GC particles were partially absorbed at the surface. HAC was in contact with partially mineralized extracellular matrix 10 weeks after implantation. In TCPC-implanted specimens, randomly oriented mineral was observed 10 weeks after implantation; however, collagenous extracellular matrix rarely was observed. In 25-week specimens, AW-GC particles were completely absorbed and replaced by new bone, and there was no intervening soft tissue. Both HAC and TCPC were in contact with bone at 25 weeks. These results indicate that AWC has higher bioactivity than either HAC or TCPC.

Animals↗
PubMed2001

Serum cystatin C may predict the prognostic stages of patients with IgA nephropathy prior to renal biopsy.

The relationship between the levels of serum cystatin C and the prognostic stages of IgA nephropathy was determined in a multicenter trial in Japan. The levels of serum cystatin C in patients with IgA nephropathy were measured using the Dade Behring N Latex Cystain C assay. In 1995, the Joint Committee of the Special Study Group on Progressive Glomerular Diseases, Ministry of Health and Welfare of Japan, and the Japanese Society of Nephropathy reported four prognostic stages. These are: good prognosis group (Group I), relatively good prognosis group (Group II), relatively poor prognosis group (Group III), and poor prognosis group (Group IV), for this disease. Three-hundred and six patients with IgA nephropathy and other glomerular diseases were examined. There were no significant changes in the levels of serum creatinine (Cr) or creatinine clearance (CCr) between Group I and Group II. The mean levels of serum cystatin C in Group II were significantly higher than those in Group I (P < 0.05). The mean levels of serum cystatin C in Group III or IV were significantly higher than those in Group I (P < 0.001, P < 0.005, respectively). These suggest that the measurement of serum cystatin C may predict the prognostic stages of patients with IgA nephropathy prior to renal biopsy.

Adult↗
PubMed2001

Hereditary angioedema with a de novo mutation of exon 8 in the C1 inhibitor gene showing recurrent edema of the hands around the peripheral joints: importance for the differential diagnosis of joint swelling.

We describe a patient with hereditary angioedema (HAE), showing recurrent edema around the peripheral joints. Her symptoms began at the age of 18 with hand swelling distal to the wrist joints. Until she was referred to our hospital 3 years after her initial symptoms, she was still undiagnosed, although she was suspected of having rheumatoid arthritis. Laboratory examination showed reduced levels of CH50 and C4 with normal C3 levels. The C1 inhibitor (C1-INH) was decreased to 5 mg/ml, with remarkably reduced activity. Although these findings were compatible with a diagnosis of HAE, there were no episodes of skin edema in her family. To establish the diagnosis, we carried out DNA analysis of the C1-INH gene, which revealed a newly identified de novo mutation of G to A at nucleotide 16869 in exon 8. As described in this patient, localized edema around the peripheral joints may be the only manifestation of HAE. HAE should therefore be taken into consideration for the differential diagnosis of joint swelling.

Adolescent↗
PubMed2001

Development of the shields for tendon injury repair using polyvinyl alcohol--hydrogel ( PVA-H).

In recent years, marked advances have been made in repair techniques for tendon injury, but the treatment of finger flexor tendon injury is still one of the most difficult and important problems in the orthopedic field. The main problem in tendon repair is adhesion between the tendon and surrounding tissue. To prevent this adhesion and achieve tendon union, we developed adhesion preventive shields for tendon repair using polyvinyl alcohol hydrogel ( PVA-H) with 90% water content, and carried out an implant experiment using the deep flexor tendon of the third toe of domestic fowl. Injured tendons shielded with PVA-H showed union at about 3 weeks after the operation without adhesion to the surrounding tissue and good function such as gliding and range of motion. Neither breakage of the PVA-H shield itself nor infection, nor degeneration in the surrounding tissue were observed. These results confirmed that the tendon itself has repair ability, and the tendon is regenerated by synovial nutrition through PVA-H. High water content PVA-H may have clinically potential and be applicable to adhesion-preventive shields for tendon repair. However, re-rupture was observed, probably due to accidental tendon injury at an early period after the operation. In some cases, tendon immobilization methods to prevent re-rupture might be necessary.

Animals↗
PubMed2001

Risk factors of hepatitis C virus-related liver cirrhosis in young adults: positive family history of liver disease and transporter associated with antigen processing 2(TAP2)*0201 Allele.

The aim of this study was to clinically characterize young patients with hepatitis-C-related cirrhosis. We compared 27 patients with liver cirrhosis (Group LC) who were anti-HCV positive, aged 40 years or less at the time of diagnosis, with 323 consecutive patients with HCV-related chronic hepatitis (Group CH) matched for age and gender. Furthermore, Group LC was divided into two arbitrary groups (29-35 years, n = 8 /36-40 years, n = 19), based on the age of patients at the time of diagnosis of liver cirrhosis. Patients' characteristics and family history were investigated, and the frequency of transporter associated with antigen processing 2 (TAP2) was determined. A family history of liver disease was present in 40.7% of Group LC but in 18.0% of Group CH (P < 0.05). The younger the age of diagnosis of cirrhosis in Group LC, the higher the frequency of a positive family history (29-35 years, 87.5%; 36-40 years, 21.1%, P < 0.05). The frequency of TAP2*0201 was significantly higher in young adult patients with HCV-related liver cirrhosis than in HCV carriers with normal ALT (P < 0.05), and tended to be higher than in uninfected normal subjects (P = 0.05). The cumulative survival rate of cirrhosis patients with family history of liver diseases was significantly lower than that of cirrhosis patients without such history (P < 0.05). Our findings suggest that a positive family history of liver disease and TAP2*0201 polymorphism may be risk factors for HCV-related liver cirrhosis in young adults.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2001

Comparison of argon-laser welded and sutured repair of inferior vena cava in a canine model.

BACKGROUND AND OBJECTIVE: Advantages of laser-welded microarterial anastomoses have been reported. However, whether laser repair of large veins is advantageous is not yet known. Argon-laser welding of inferior vena cava was therefore compared with conventional-sutured repair. STUDY DESIGN/MATERIALS AND METHODS: Twenty-four adult mongrel dogs were used. Twelve inferior vena cavas were repaired using an argon laser and the rest using continuous sutures. Specimens were removed at 2, 8, 16, and 24 weeks after repair. Technical characteristics, cavographical findings, and results of histopathological study using light and transmission electron microscopy were compared. RESULTS: Procedure time did not significantly differ between the two groups. The sutured repair sites were narrower than those repaired by laser immediately after and at 2 and 24 weeks. Histologically, laser welding resulted from denaturation of collagen and mild healing occurred simultaneously with the disappearance of denatured collagen. By contrast, there was strong fibrotic reaction at sutured repair sites. CONCLUSIONS: These findings suggest that argon-laser repair of large veins is superior to continuous suturing.

Anastomosis, Surgical↗
PubMed2001

Studies on the association of 2-thiazolidinecarboxylic acid and antimony potassium tartrate: chiral recognition and prediction of absolute configuration by electrospray ionization mass spectrometry.

Optically active 2-thiazolidinecarboxylic acid (2-THC), a substrate for D-amino acid oxidase in animal kidney, is known to undergo racemization quickly in solution. The association of (+)- and (-)-2-THC with antimony potassium tartrate K(2)[Sb(2)(L or D-tart)(2)] was studied by electrospray ionization mass spectrometry (ESI-MS). We observed that relative intensities of associated ions in acetonitrile/water solution were changing as the racemization progressed. For [Sb(2)(L-tart)(2)](2-), the intensities of the associated ions increased as (+)-2-THC underwent racemization to a (-)-isomer; on the other hand, the intensity of the associated ion decreased as (-)-2-THC underwent racemization to a (+)-isomer. In the case of [Sb(2)(D-tart)(2)](2-), an opposite effect on the intensities of the associated ions was observed. The change in the intensities of associated ions can be used for chiral recognition of (+)-2-THC and (-)-2THC. Stereochemical models of the association of the optical isomers with [Sb(2)(L- or D-tart)(2)](2-) were constructed from the consideration of both hydrogen bonding of NH-O functions and HSAB (hard and soft acids and bases) interaction of S and Sb atoms. Comparison of the stereochemical models with the ESI-MS results enabled us to predict the absolute configurations of the 2-THC isomers.

Journal Article↗