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Biomedical subjects

M Takiguchi

Publications and source records attributed to M Takiguchi.

At least 109 records · Page 6Linked to original sources

The effect of epitope variation on the profile of cytotoxic T lymphocyte responses to the HIV envelope glycoprotein.

To address the relationship between viral and host factors during HIV infection, we analyzed the effect of viral mutations on T cell responses in seropositive, asymptomatic HLA-A2+ individuals using four envelope (env)-specific peptides with the HLA-A*0201 binding motif. We showed that the natural sequence variation was frequent within epitopes located in the C-terminal region of the env glycoprotein and was largely responsible for a lower env-specific cytotoxic T lymphocyte (CTL) activity in the peptide-stimulated cultures. The highest CTL responses in vitro were induced with conserved epitopes D1 and 4.3 that mapped to the N-terminal region of the env glycoprotein. These peptides exhibited high binding affinity for HLA-A*0201 molecules and stimulated CD8+ T cells of relatively limited TCR Vbeta chain repertoire. Decreased CTL activities to the D1 epitope were observed in the absence of any detectable viral mutation, and were associated with lower proliferative responses and expression of the CD28 antigen. Results of this study demonstrate that the degree of sequence variation within a stimulatory epitope of the viral quasispecies, as well as proliferative potential of the effector cells, are among the factors underlying decreased CTL activity in HIV-infected patients. These experiments also provide evidence that the D1 peptide might be useful for the development of vaccines and immune-based therapy.

Adult↗

Identification and characterization of a cytotoxic T cell epitope of hepatitis C virus presented by HLA-B*3501 in acute hepatitis.

In order to clarify the role of cytotoxic T lymphocytes (CTL) in hepatitis C virus (HCV) infection, an HLA-B35-restricted cytotoxic T cell epitope of HCV was identified using a strategy called reverse immunogenetics. Twenty-eight of 53 HCV peptides carrying two anchor residues were selected as HLA-B*3501 binding peptides. These peptides were used to induce the specific cytotoxic T cells in peripheral blood lymphocytes from a patient with acute hepatitis C. Six HLA-B*3501 binding peptides induced the peptide-specific CTL. One (HPNIEEVAL) of five peptides was confirmed as the epitope by the specific T cell clones. A sequence identical to the epitope was detected in isolates of the virus from the patient and a strong CTL response to this epitope was observed in the acute phase of hepatitis C but not in the recovery phase. The specific CTL for this epitope were not detected in peripheral blood lymphocytes from patients with chronic hepatitis C. Together these results suggest that the CTL specific for this epitope have an important role in the elimination of the virus in the patient.

Acute Disease↗

The role of anchor residues in the binding of peptides to HLA-A*1101 molecules.

The binding of 136 8- to 12-mer peptides carrying anchor residues at position 2 (P2) and the C-terminus to HLA-A*1101 molecules was analyzed by a stabilization assay using RMA-S transfectants expressing HLA-A*1101 and human beta2-microglobulin. 72.1% of these peptides bound to HLA-A*1101 molecules. Two known HLA-All-restricted cytotoxic T-lymphocyte epitope peptides showed high affinity to HLA-A*1101. The results confirmed a previous pool sequencing study of HLA-A*1101 binding self-peptides, which showed that Lys at the C-terminus and Val, Ile, Phe, Tyr, and Thr at P2 are anchor residues for HLA-A*1101. Thr and aliphatic hydrophobic residues Val, Ile, and Leu at P2 are stronger anchor residues than the aromatic hydrophobic residues Phe and Tyr. In addition, hydrophobic residues Leu, Phe, Tyr, Ile, and Ala at position 3 (P3) are secondary anchors but are weaker than those at P2. The affinities of the 8- and 12-mer peptides were significantly lower than those of 9- to 11-mer peptides. There was however no difference in affinity between 9-, 10- and 11-mer peptides. Furthermore, the analysis using peptides mutated at the C-terminus showed that HLA-A*1101 molecules can bind peptides carrying another positively charged residue, Arg. The present study clarified the role of the anchor residues at P2, P3 and the C-terminus in the binding of HLA-A*1101 molecules.

Binding Sites↗

The advantage of magnetic resonance imaging in diagnosis of cauda equina syndrome in dogs.

Three dogs were evaluated in our study using magnetic resonance imaging (MRI) to reveal the anatomical deformity and the degree of the lesion of cauda equina. In all dogs, MRI revealed soft tissue, such as cauda equina, epidural fat, and intervertebral disc, at the lumbosacral region clearly without contrast medium. Our results suggest that MRI has some advantages in evaluating cauda equina syndrome in dogs.

Animals↗

[Two cases of complete atrioventricular canal defect whose pulmonary vascular resistance were over 10 wood unit.m2 before 6 months of age].

We report two cases with complete form of atrioventricular canal defect (CAVCD) accompanied by Down's syndrome whose pulmonary vascular resistance (Rp) were more than 10 Wood unit.m2 at the age of less than 6 months. One child was 3-month-old boy whose Rp was 12 Wood unit.m2. The open lung biopsy at 3 months old showed histopathological change of Heath-Edwards grade I. He underwent intracardiac repair at the age of 4 months. He is doing well at 30 months of postoperative period. Another child was 5-month-old girl whose Rp was 15.5 Wood unit.m2. Histopathological change of lung at 5 months old demonstrated Heath-Edwards grade III. She underwent intracardiac repair at the age of 7 months, 2 months after lung biopsy. However, she died of oversystemic pulmonary hypertension and low output syndrome 7 days after surgery. The postmortem examination revealed that pulmonary vascular obstructive disease progressed during 2 months interval between the lung biopsy and the operation. In conclusion, cardiac catheterization with estimation of Rp should be performed in the cases of CAVCD, especially in those with Down's syndrome, in early infancy. If Rp is more than 10 Wood unit.m2 and lung biopsy indicates the surgical indication, surgical intervention should be done as soon as possible since the pulmonary vascular obstructive disease may progress in a short period.

Endocardial Cushion Defects↗

The canine alkaline phosphatases: a review of the isoenzymes in serum, analytical methods and their diagnostic application.

This paper reviews the alkaline phosphatases in canine serum, the analytical methods used for qualitative and/or quantitative detection of these isoenzymes, and the diagnostic significancy of each of these isoenzymes. The paper further describes some of the latest advances in our knowledge of the canine alkaline phosphatases and possible areas of future research.

Alkaline Phosphatase↗

[A surgical case of angina pectoris with a severe stenosis of Lt. mid-cerebral artery: the usefulness of the monitoring of cerebral blood flow].

A 64-year-old male patient had two episodes of transient ischemic attack and a cerebral infarction. Cerebral angiography showed 50% stenosis at the junction of left internal carotid artery and 90% stenosis at left mid-cerebral artery (MCA). Coronary angiography showed two vessel disease with arteriosclerotic change and underwent coronary artery bypass grafting. To prevent intraoperative cerebral infarction, we used brain protect solution just before starting ECC, set perfusion flow around 3 l/min/m2, monitored the flow of left MCA using Transcranial Doppler (TCD) and the saturation of left internal jugular vein (SjO2) continuously. PaCO2 was controlled around 45 mmHg. TCD showed good pulsatile flow, and SjO2 was kept over 60%. The patient recovered consciousness 2 hours after operation in the intensive care unit without paresthesia. We thought the number of open-heart cases with cerebrovascular disease increased, and pulsatile low of ECC by intraaortic balloon pumping and the monitoring of SjO2 are useful for the cases.

Angina Pectoris↗

[Cases of multiple ventricular septal defects].

We had 4 cases with multiple ventricular septal defects (VSDs) in complexed congenital heart disease. One of four had two separate VSDs detected by two dimensional echocardiography before operation. Second of four had additional infundibular muscular VSD which was detected by echocardiography in the intensive care unit (ICU) after patch closure of a perimembranous VSD. The third case had two additional VSDs of inlet muscular and subaortic septum detected by transesophageal and direct echocardiography during reoperation, beside a subpulmonary VSD which was originally diagnosed before Jatene operation for double outlet right ventricle. The fourth case had multiple trabecular muscular VSDs diagnosed by postoperative angiography soon after Rastelli operation. Since these additional multiple VSDs compromise the postoperative hemodynamics if those are unrecognized, it is indispensable to detect all VSDs before operation, using transthoracic and transesophageal echocardiography.

Echocardiography↗

Massive aortic regurgitation by spontaneous rupture of a fibrous strand in a fenestrated aortic valve.

A very rare case of Marfan syndrome associated with acute aortic regurgitation due to rupture of a fibrous strand of a fenestrated aortic valve is presented. Although an aortic valve fenestration usually has no pathologic significance, its association with aortic annular dilatation plays an important role. Rupture of a myxomatously degenerated fibrous strand loaded with high tension from annular dilatation reducing the coaptation area of each cusp during diastole was the suspected cause of acute massive regurgitation by complete retroversion of the right coronary cusp. Rupture of a fibrous strand of a fenestrated aortic valve with a dilated annulus could result in acute massive aortic regurgitation, for which prompt surgical repair is mandatory.

Adult↗

[Role of the home health care/visiting nurse in industry: possibilities for visiting nurse station and industry cooperation].

As societal awareness of home health care increases, patient demands for home health care are becoming more diversified. In the future, the construction of a system able to address this diversification will be essential for further establishing the role of home health care in society. A demand exists for hospital primary care physicians, medical practitioners, visiting nurse stations, helper services, and firms to provide their services in flexible combination with those of organizations and occupations having various functions, in forms that suit the needs of the relevant patient group. Particularly in the case of visiting nursing, the cultivation of internal flexibility and cooperative relationships with outside organizations able to serve a variety of needs may prove to be the essential lesson remaining after the discontinuation of care insurance, two years after its introduction. Through cases involving physicians, visiting nurse stations, and firms we consider favorable ways for organizations to cooperate.

Aged↗

Functional association between CBP and HNF4 in trans-activation.

CBP functions as a key transcriptional coactivator for a variety of transcription factors. We show here that the hepatocyte nuclear factor 4 (HNF4), a transcription factor in the nuclear receptor superfamily with no defined ligand, is cloned by yeast two-hybrid system using CBP as a bait. GST-pull down assay with nuclear extracts or in vitro translation products revealed that CBP and HNF4 interact with each other at the middle portion (aa 119-375) of HNF4 and two distinct regions (aa 271-451 and 1626-2259) of CBP, respectively, in the ligand-independent manner. Co-transfection experiments indicated that CBP is capable of activating HNF4 site-mediated transcription. These results suggested a functional association between CBP and HNF4 in trans-activation.

Animals↗

Identification and characterization of multiple HLA-A24-restricted HIV-1 CTL epitopes: strong epitopes are derived from V regions of HIV-1.

HIV-1-specific CTL has a crucial role in the elimination of the virus. However, a restricted number of common HLA class I alleles has been studied for their presentation of HIV-1 CTL epitopes. We have attempted to identify HIV-1 CTL epitopes presented by HLA-A*2402 using reverse immunogenetics. Fifty-three HLA-A*2402-binding HIV-1 peptides were used to induce specific CTL in PBL of four HIV-1-infected individuals carrying HLA-A24. Twelve peptides were strongly suggested to be HLA-A*2402-restricted HIV-1 CTL epitopes because these peptides induced the specific CTL that killed both the target cells pulsed with the specific peptides and those infected with the vaccinia HIV-1 recombinant virus in at least one HIV-1-infected individual. Of these epitopes, 11 were confirmed by the generation of the specific CTL clones. Six were the Env epitopes while three, one, and one were derived from Gag, Pol, and Nef proteins, respectively. Analysis of 12 HIV-1-infected individuals using these peptides showed that 5 derived from the Env protein and one from the Nef protein were strong epitopes. These strong epitopes were derived from the diverse region of HIV-1 while weak epitopes were conserved in the HIV-1 clade B strain. Analysis of CTL recognition of mutations in these strong epitopes suggested that the mutations in the Env epitopes may critically influence CTL recognition in vivo.

Alleles↗

Evidence of presentation of multiple HIV-1 cytotoxic T lymphocyte epitopes by HLA-B*3501 molecules that are associated with the accelerated progression of AIDS.

We recently showed HLA-B35-restricted CTL activity for 10 HIV-1 epitopes in PBL from two HIV-1-infected individuals. In the present study, we established CTL clones specific for nine of these HIV-1 epitopes to confirm these HLA-B35-restricted epitopes. The specific CTL clones effectively killed the HLA-B*3501-positive target cells infected with the HIV-1 vaccinia recombinant virus. These results confirmed that nine HIV-1 CTL epitopes are presented by HLA-B*3501 molecules. The CTL activity specific for four Pol and two Nef epitopes was induced in the peptide-stimulated PBL from three or more of seven HIV-1-infected individuals, indicating that these six are common epitopes. Eight were considered strong epitopes because the specific CTL activity was detected in the cultured PBL that was once stimulated with peptides. Thus, the present study excluded the possibility that the disability of the presentation of HIV-1 epitopes by HLA-B35 molecules is associated with the accelerated progression of AIDS in HLA-B35-positive individuals. Analysis of mutated epitopes found in an HIV-1 type B strain using the CTL clones revealed that most mutated epitopes partially or markedly affect the recognition of CTL clones. Of 19 mutations that affected recognition of the CTL clones, 7 reduced peptide-HLA-B*3501 binding, while 12 affected TCR recognition. These results indicate that natural mutations of HLA-B35-restricted HIV-1 CTL epitopes affect the recognition of CTL by mechanisms that reduce both peptide binding and TCR recognition.

Acquired Immunodeficiency Syndrome↗

Coinduction of nitric-oxide synthase and arginase I in cultured rat peritoneal macrophages and rat tissues in vivo by lipopolysaccharide.

Nitric oxide is synthesized by nitric-oxide synthase from arginine, a common substrate of arginase. Rat peritoneal macrophages were cultured in the presence of bacterial lipopolysaccharide (LPS), and expression of the inducible isoform of nitric-oxide synthase (iNOS) and liver-type arginase (arginase I) was analyzed. mRNAs for iNOS and arginase I were induced by LPS in a dose-dependent manner. iNOS mRNA appeared 2 h after LPS treatment and increased to a near maximum at 8-12 h. On the other hand, arginase I mRNA that was undetectable prior to the treatment began to increase after 4 h with a lag time and reached a maximum at 12 h. Immunoblot analysis showed that iNOS and arginase I proteins were also induced. mRNA for arginase II, an arginase isozyme, was not detected in the LPS-activated peritoneal cells. mRNA for CCAAT/enhancer-binding protein beta (C/EBPbeta), a transactivator of the arginase I gene, was also induced, and the induction was more rapid than that of arginase I mRNA. Changes in iNOS and arginase I mRNAs were also examined in LPS-injected rats in vivo. iNOS mRNA increased rapidly in the lung and spleen, reached a maximum 2-6 h after the LPS treatment, and decreased thereafter. Arginase I mRNA was induced markedly and more slowly in both tissues, reaching a maximum in 12 h. Thus, arginase I appears to have an important role in down-regulating nitric oxide synthesis in murine macrophages by decreasing the availability of arginine, and the induction of arginase I is mediated by C/EBPbeta.

Animals↗

The glucocorticoid-responsive gene cascade. Activation of the rat arginase gene through induction of C/EBPbeta.

The gene for liver-type arginase, an ornithine cycle enzyme, is induced by glucocorticoids in a delayed secondary manner. An enhancer element located around intron 7 of the rat arginase gene shows delayed glucocorticoid responsiveness, and it harbors two sites binding with members of the CCAAT/enhancer binding protein (C/EBP) family. Here, we investigate the role of these C/EBP binding sites in glucocorticoid response of the arginase gene. When inserted in front of the herpes simplex virus thymidine kinase promoter, these C/EBP sites exhibited glucocorticoid responsiveness in reporter transfection assay using rat hepatoma H4IIE cells. In footprint analysis using nuclear extracts of H4IIE cells, profiles of the protected areas of the two C/EBP sites changed when cells were treated with dexamethasone. In gel shift analysis, the complex formation for the two C/EBP sites was augmented in response to dexamethasone. Antibody supershift/inhibition analysis demonstrated that a major portion of the binding proteins induced by dexamethasone is C/EBPbeta. Induction of arginase mRNA by dexamethasone was preceded by augmentation of the C/EBP site-binding activities, which followed increase in C/EBPbeta mRNA. These results were consistent with the notion that the glucocorticoid response of the arginase gene is mediated by C/EBPbeta.

Animals↗

The release of nitroglycerin absorbed into the central venous catheter.

This study was conducted to evaluate the release of nitroglycerin (NG) that has been absorbed into the central venous catheter. A 0.05% NG solution was infused through a central venous catheter and the flow rates were set at 1, 5, or 10 ml/h, given over 12, 24, or 48 h. The catheter was flushed with lactate Ringer solution after completion of the NG infusion. The elution of the lactate Ringer solution from the tip of the catheter was then collected and assayed for its NG concentration by high performance liquid chromatography (HPLC). A higher concentration of NG was released with a faster flow rate and a longer infusion. The high level of NG release continued during the first 20 min, and ranged from a minimum of 0.07 mg/ml to a maximum that exceeded 0.15 mg/ml. Subsequently, the NG concentration gradually declined, but low concentrations of 0.006-0.02 mg/ml were still maintained 360 min later. Thus, it is suggested that if a catheter such as the Swan-Ganz continues to be used after the completion of a NG infusion, certain pharmacological effects due to the absorption of NG into the catheter body should be expected for at least 60 min.

Absorption↗

Binding of 8-mer to 11-mer peptides carrying the anchor residues to slow assembling HLA class I molecules (HLA-B*5101).

The binding of 303 8-mer to 11-mer peptides carrying the anchor residues at P2 and the C-terminus to HLA-B*5101 molecules was examined by a stabilization assay in which peptides were incubated with RMA-S-B*5101 cells at 26 degrees C for 3 h. Analysis of the binding of these peptides to HLA-B*5101 molecules showed that Pro and Ala at P2, and Ile, Val, and Leu at the C-terminus functioned as anchor residues, while Gly at P2 and Met at the C-terminus were weak anchors. Pro was a stronger anchor residue than Ala at P2, while Ile was the strongest anchor at the C-terminus. Among 8-mer to 11-mer peptides, the 9-mer peptides showed the strongest binding to HLA-B*5101 molecules. This is in contrast to our recent findings that 10-mer and 11-mer peptides bind to HLA-B*3501 molecules as effectively as 9-mer peptides. Since both HLA class I molecules have the same B-pocket and the binding peptides carry the same anchor residues, it is assumed that the structure of the F-pocket may restrict the length of binding peptides. The ability of HLA-B*5101 binding peptides to stabilize the HLA-B*5101 molecules was markedly lower than that of HLA-B*3501 binding peptides to stabilize the HLA-B*3501 molecules. It is known that HLA-B*5101 is a slow assembling molecule, while HLA-B*3501 assembles rapidly. The results imply that the slow assembling of HLA-B*5101 molecules results from the low affinity of peptides to HLA-B*5101 molecules.

Cell Line↗