Cryopreserved femoral arterial allografts for vascular access in hemodialysis.
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Biomedical subjects
Publications and source records attributed to T Tsuji.
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Collagen, as a component of the extracellular matrix, have a role in the healing process after myocardial infarction (MI). For type IV collagen, a major structural protein present in the basal membrane of myocytes, six alpha chains [alpha 1 (IV)-alpha 6(IV)] have been identified. We examined the sequential changes in the appearance and localization of the alpha 1 (IV)-alpha 5(IV) after experimental MI in rats. Hearts were excised from 1 day to 8 weeks after permanent left coronary artery ligation. Immunohistochemical staining with monoclonal antibodies was performed. On day 3, staining for both alpha 1(IV) and alpha 2(IV) first appeared, forming a wavy pattern in the infarct peripheral zone, and the staining was not restricted to the cell membrane. The staining intensity and distribution for both alpha 1(IV) and alpha 2(IV) in the peripheral zone then gradually increased, reaching a maximum around day 7. The distribution progressed from the peripheral to the central zone of the infarct for 1-2 days, reaching the center point after 2 weeks. The staining distribution gradually decreased after reaching the maximum, but the staining had not completely disappeared at 8 weeks. In contrast, no positive staining for alpha 3(IV), alpha 4(IV) or alpha 5(IV) was observed at any time during the 8-week observation period. Thus, the present results demonstrated that in rats, type IV collagen consisting of alpha 1 and alpha 2 chains appears in the infarct zone at a relatively early phase after MI, indicating that type IV collagen composed of alpha 1 and alpha 2 chains contributes to infarct healing.
Enhancement of DNA vaccine immunogenicity is a current topic of high priority in the field of applied immunology, especially as a means of controlling HIV infection. The adjuvant effect of Ubenimex (UBX), an anti-cancer immunomodulator, on a DNA AIDS vaccine which we developed was examined in a murine model. UBX was formulated into a preparation containing DNA plasmids encoding env and rev genes of HIV-1 strain III(B), and was inoculated intramuscularly into BALB/c mice. The sera obtained with this mixture had 2(3)-2(5) times higher specific IgG titres than those obtained without the use of the adjuvant. UBX also elicited both a stronger HIV-1-specific DTH reaction, as measured by the footpad swelling test, and stronger cytotoxic T lymphocyte activity, as assayed by the 51Cr-release method, compared with responses using DNA alone. The cytokine secretion profile of restimulated immune lymphoid cells showed that UBX raised IL-2 and interferon-gamma levels and decreased IL-4 production. HIV-1-specific immunoglobulin subtype analysis demonstrated that UBX stimulated IgG2a production but suppressed synthesis of IgG1 and IgE. These results indicate that activation of the T-helper type 1 subset was induced by UBX, suggesting a mechanism of immunomodulation mediated by this agent. We conclude that UBX acts as an immunologic adjuvant for DNA vaccination against HIV-1. UBX may be a suitable adjuvant for clinical use because of its lack of antigenicity and low toxicity.
Colonic epithelial cells of patients with UC express DAF in relation to the severity of mucosal inflammation. The aim of this study was to determine whether this factor in stool could be used as a marker of disease activity in UC patients. Stool DAF was measured by use of an immunoassay in 181 stool specimens obtained from 55 patients with UC of various levels of disease activity. Stool DAF concentrations in patients whose UC was active (0.0-785.6 ng/g stool; median 47.1 ng/g; n = 115) were significantly higher than concentrations in patients whose disease was inactive (0.0-48.6 ng/g; median 0.0 ng/g; n = 66) (P < 0.0001). Values in active UC patients also were higher than those in control patients with diarrhoea (0.0-30.0 ng/g; median 0.0 ng/g; n = 26) (P < 0.0001) and in control subjects without apparent colorectal disease (0-20.4 ng/g; median 0.0 ng/g; n = 44) (P < 0.0001). The elevated levels of stool DAF obtained from UC patients in relapse declined markedly in specimens collected after the disease went into remission following medical therapy. Stool DAF levels correlated with the severity of endoscopic and histological findings and the degree of DAF expression on the colonic epithelia. Our results suggest that the measurement of stool DAF is useful as a non-invasive means of monitoring intestinal disease activity in patients with UC.
Expression of DAF (CD55) is enhanced on colonic epithelial cells of patients with ulcerative colitis (UC), and stool DAF concentrations are increased in patients with active disease. Cytokines are known to modulate DAF expression in various human cells, and lesions of UC reveal altered profiles of cytokine production. In this study, we evaluate the effects of various cytokines, IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, and interferon-gamma (IFN-gamma), on the synthesis and kinetics of DAF protein in HT-29 human intestinal epithelial cells. Using flow cytometry and an ELISA, we found that HT-29 cells constitutively express DAF on the cell surface and spontaneously release DAF into the culture supernatant under standard culture conditions. When the culture supernatant was centrifuged at 100000g, nearly a half of DAF was precipitated, indicating that one half of the released DAF was present as a membrane-bound form and the other half as a soluble form. Analysis of the culture supernatant of biotin surface-labelled HT-29 cells suggested that the soluble form DAF was derived by secretion from within the cell or by cleavage from the cell surface. Among the cytokines, IL-4 markedly, and IL-1beta moderately, enhanced the expression and the release of DAF. Actinomycin D, cycloheximide, and brefeldin A inhibited the increase in DAF release induced by IL-4 and IL-1beta stimulation. These results suggest that DAF is released from intestinal epithelial cells in response to cytokine stimulation and that IL-4 and IL-1beta are possible cytokines involved in DAF release into the colonic lumen of patients with UC.
DNA vaccine against human immunodeficiency virus type-1 (HIV-1) can induce substantial levels of HIV-1-specific humoral and cell-mediated immunity. To develop more potent HIV-1 DNA vaccine formulations, we used a murine model to explore the immunomodulatory effects of an interleukin-2 (IL-2) expression plasmid on an HIV-1 DNA vaccine following intranasal administration of the combination. When the vaccine and expression plasmid were incorporated into cationic liposomes and administered to mice, the HIV-1-specific delayed-type hypersensitivity response and cytotoxic T lymphocyte activity were significantly increased. Restimulated immune lymphoid cells showed enhanced production of both IL-2 and interferon-gamma and reduced secretion of IL-4. The level of total antibody to HIV-1 antigen was not greatly changed by coadministration of the DNA vaccine and IL-2 expression plasmid. An analysis of serum HIV-1-specific IgG subclasses showed a significant drop in the IgG1/IgG2a ratio in the group that received the plasmid-vaccine combination. These results demonstrate that the IL-2 expression plasmid strongly enhances the HIV-1-specific immune response via activation of T helper type-1 cells.
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Gc globulin, also called vitamin D-binding protein, is a human 56 kDa plasma protein. Antigenic epitopes of the protein recognized by 12 different monoclonal antibodies (MoAbs), including the previously prepared MoAb E12 of which binding to Gc is inhibited by actin, were analyzed in relation to the functional domains of the protein. To map the epitopes recognized by the respective antibody, the reactivities of the MoAbs were tested by immunoblotting against the recombinant Gc fragments expressed in E. coli cells, and then a competitive ELISA was performed. The results showed that the antibodies were classified into at least six groups. Furthermore, in addition to E12, actin inhibited the reactivity of MoAb 21, of which the epitope was located within residues Asp320 to Glu379, in an ELISA in the presence of the ligand. This result must indicate that this antibody-binding site is near the site that interacts with actin. In contrast, no inhibitory effect by 25-hydroxyvitamin D to any antibodies was observed. Furthermore, all antibodies, including E12 and 21, reacted with membrane-bound Gc of lymphocytes by an immunofluorescence study, suggesting that Gc is unlikely to bind to the plasma membrane in its interaction with actin.
The administration of ABO blood group active glycoproteins via the rear footpads of BALB/c mice with subsequent fusion of popliteal lymph node lymphocytes produced hybridomas differentially secreting antibodies to the peptide moiety of antigens. On the other hand, conventional intraperitoneal immunization of the same antigens and intraperitoneal boost followed by splenic lymphocyte fusion produced hybridomas differentially secreting antibodies to the carbohydrate moiety of antigens. Similar results were obtained in the production of monoclonal antibodies to erythrocyte membranes. Almost all hybridomas obtained by rear foodpad immunization secreted antibodies to the peptide moiety of erythrocyte membrane proteins, and those obtained by intraperitoneal immunization secreted antibodies to both peptide and carbohydrate moieties of erythrocyte membrane components. These results suggest the possibility of differential production of monoclonal antibodies to the carbohydrate moiety and the peptide moiety of some glycoproteins by different immunization routes.
We semiquantitatively analyzed expression of PCNA and LeY in seborrheic keratosis (SK), actinic keratosis (AK), Bowen's disease (BD), and squamous cell carcinoma (SCC), using immunocytochemically stained tissue sections. PCNA expression increased in a stepwise fashion from low levels in normal skin to higher expressions within SK, AK, BD, and SCC. The levels of LeY protein also increased in this order. The PCNA expression pattern shifted from expression limited to the basal and suprabasal cell layers (in normal skin and SK) to expression extending to the upper squamous and granular layers (in AK, BD, and SCC). On the other hand, the pattern of LeY expression shifted from the granular (in normal skin) to the upper squamous (in SK and AK) and suprabasal layers (in BD and SCC). These findings suggest that PCNA expression is related to the degree of cell proliferation and that LeY expression is related to the degree of differentiation or keratinization of tumor cells. In addition, PCNA and LeY show a reciprocal relationship in their expression.
Sclerosing stromal tumor (SST) is a rare ovarian neoplasm occurring predominantly in young women and is histologically characterized by cellular heterogeneity, prominent vasculature, and a pseudolobular appearance composed of cellular and hypocellular areas. In the current study, three cases of SST were ultrastructurally examined and the tumors were found to be composed of several kinds of cells, i.e., luteinized thecalike cells, spindle-shaped fibroblastic cells, and primitive mesenchymal cells. These findings thus supported the ovarian stromal origin of SST. Twelve cases of SST also were analyzed immunohistochemically and demonstrated an expression of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) in the luteinized thecalike cells and its receptor, fms-like tyrosine kinase 1 (flt-1), in capillary to medium-sized blood vessels. Reverse transcription-polymerase chain reaction (RT-PCR) also showed an expression of VPF/VEGF messenger RNA in SSTs. Accordingly, the characteristic vasculature and edema of SSTs were considered to be associated with the expression of VPF/VEGF. In addition, a fluorescence in situ hybridization (FISH) analysis also showed cells with three copy number of chromosome 12 in 13-21% of all examined SST cells, which suggested the presence of chromosome 12 trisomy in SSTs as well as in other ovarian stromal tumors.
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OBJECTIVE: Oxyhemoglobin (OxyHb) is thought to be a critical trigger in the pathogenesis of cerebral vasospasm after aneurysmal subarachnoid hemorrhage. We investigated whether extraluminally applied OxyHb influenced vascular responses to intraluminally applied vasoactive agents in isolated, perfused, canine basilar arteries. METHODS: The steel cannula insertion method was used to examine vascular responses to intraluminally applied 5-hydroxytryptamine (5-HT) receptor agonists, i.e., 5-HT, 5-carboxamidotryptamine (selective for 5-HT1 receptors), and alpha-methyl-5-hydroxytryptamine (selective for 5-HT2 receptors), potassium chloride, and acetylcholine, before and after extraluminal treatment with OxyHb. RESULTS: Extraluminal application of 2.5 x 10(-5) mol/L OxyHb immediately induced a transient elevation of the basal perfusion pressure, which gradually decreased and then stabilized at a level slightly higher than the control level. Each 5-HT agonist induced dose-dependent vasoconstriction. The potencies of the agonists were not very different, but the efficacies varied, i.e., alpha-methyl-5-hydroxytryptamine > 5-HT > 5-carboxamidotryptamine. Each response was strongly inhibited by ketanserin (a selective 5-HT2 receptor antagonist), indicating that each agonist induces vasoconstriction mediated by 5-HT2 receptors. The vasoconstriction in response to each 5-HT receptor agonist was consistently potentiated by treatment with OxyHb (2.5 x 10(-5) mol/L). 5-HT receptor agonist-induced constrictions after OxyHb treatment were much more markedly inhibited by ketanserin, compared with those before OxyHb treatment. Acetylcholine-induced constrictions were enhanced by OxyHb, but KCl-induced constrictions were significantly decreased by OxyHb. CONCLUSION: It is suggested that OxyHb enhancement of constrictions in response to 5-HT receptor agonists may be mediated by increased sensitivity of 5-HT2 receptors, in addition to actions in the endothelium, in canine basilar arteries. This potentiated vasoconstrictor mechanism may be partially implicated in cerebral vasospasm after subarachnoid hemorrhage.
We report a case of pustular drug eruption due to bacampicilin hydrochloride which developed in a patient with pustular psoriasis. The patient was a 45-year-old Japanese woman with psoriasis which started as pustular psoriasis twenty years previously. In 1994, she developed generalized erythema with pustules accompanied by high fever and liver injury. Clinical and histological findings of this pustular eruption were different from her previous episodes of pustular psoriasis. Erythemas and pustules disappeared and her abnormal transaminase returned to normal rapidly when she discontinued bacampicilin hydrochloride. Her positive reaction to a patch test and a lymphocyte stimulation test also suggested that our case had a pustular drug eruption rather than pustular psoriasis induced by a drug.
BACKGROUND: Familial prostate cancer has been studied in Europe and the United States. This study was conducted to clarify the clinical features and incidence of the human leukocyte antigen (HLA) in familial prostate cancer in patients seen at hospitals in Japan. METHODS: The age at diagnosis, clinical stage, histologic differentiation, prognosis, cause of death and family history were compiled from 43 patients from 20 families with familial prostate cancer (group F) and compared in patients with sporadic prostate cancer (group S). In addition, HLA class II gene typing was performed in 17 patients from 11 families with familial prostate cancer (group F') and their healthy family members (15 males and 25 females). RESULTS: The age at diagnosis was significantly lower in group F patients (P< 0.001), but neither clinical stage nor prognosis differed between group F and group S, yet the incidence of moderately differentiated adenocarcinoma was significantly higher in group F (P< 0.01 ). There were no historical factors that were specific for group F patients. The frequency of HLA-DRB1 *0901 and *1201 alleles was significantly higher in group F' patients than in group J (the frequency of alleles in 1216 Japanese people attending the 11th Japan HLA Workshop), but not between group F' and their healthy male family members. CONCLUSION: There were no significant differences in clinical parameters between group F and group S, except that the age at diagnosis was lower in group F patients, and that the frequency of several HLA class 11 alleles was significantly higher in patients with prostate cancer. In men with a family history of prostate cancer, examinations for early detection of prostate cancer should begin at a young age.
We identified a family of 4 brothers with prostate cancer. The proband was the second son who was seen at the Department of Urology, Gunma University, Gunma, Japan, in May 1995, with a chief complaint of dysuria. The serum level of PSA was high, and a prostate needle biopsy revealed a moderately-differentiated adenocarcinoma. Subsequently, a total prostatectomy was performed. The family history revealed that the fourth son had been under treatment at the Saitama Cancer Center Hospital for prostate cancer since 1994. Therefore, the other 2 brothers became healthy male family members of the familial prostate cancer line, and their serum PSA levels were examined after obtaining their informed consent. Both had a high serum PSA level and biopsies revealed moderately-differentiated adenocarcinomas in both. Human leukocyte antigen class II gene typing was conducted on 3 of 4 brothers and 4 healthy family members who gave informed consent, and revealed the DRB1*0901-DQB1*0303 haplotype in the first, second and third sons. Familial prostate cancer is well documented in Europe and the US where the incidence of prostate cancer is high, but this is the first report of prostate cancer in 4 brothers in Japan.
We tried to compare the efficacy of plasma exchange (PE) with that of intravenous immunoglobulin (IVIG) in patients with postinfectious polyneuritis (Guillain-Barré syndrome [GBS] and cranial neuritis). Fifteen patients with postinfectious polyneuritis were divided into 2 groups. The IVIG group included 5 cases of GBS and 2 cases of postinfectious cranial neuritis (ophthalmoplegic type). The PE group included 5 cases of GBS and 3 cases of postinfectious cranial neuritis (ophthalmoplegic type). The changes and incidences of improvement of muscle strength scores (MSSs) and ocular movement scores (OMSs) were evaluated before treatment and 1, 2, 3, and 4 weeks after treatment. No significant differences between the IVIG and PE groups were found in the MSSs or OMSs at any time after treatment. These data suggested that PE and IVIG had equivalent efficacy. In the IVIG group, the proportion of suppressor-inducer T cells significantly increased (p < 0.01) (before versus after treatment), and the proportion of suppressor-effector cells also increased but not significantly (before versus after treatment). In the PE group, the percentage of suppressor-inducer T cells significantly decreased (p < 0.05) (before versus after treatment) while the proportion of suppressor/cytotoxic T cells significantly increased (p < 0.05) (before versus after treatment). The percentage of suppressor-effector T cells also increased (before versus after treatment) but not significantly.
In East Asian countries, the prevalence of viral hepatitis has been reported to be high, but precise data for each country remained to be investigated. Here we report the prevalence of viral markers of hepatitis B and hepatitis C in outpatient volunteers visiting two general hospitals in Ulaanbaatar, Mongolia. One hundred fifty sera were tested for HBs antigen (HBsAg), anti-HBs, and anti-HCV by Counting Immunoassay. The backgrounds of groups of patients positive for HBsAg and negative for anti-HCV (group 1; n = 18), negative for HBsAg and positive for anti-HCV (group 2; n = 47), positive for both HBsAg and anti-HCV (group 3; n = 25), and negative for both HBsAg and anti-HCV (group 4; n = 60) were compared. The prevalence of HBsAg, anti-HBs, and anti-HCV in this study group was 28.7%, 39.3% and 48.0%, respectively. Subjects of group 1 (mean +/- SD; 31.3 +/- 12.4 years old) were younger than those of group 4 (39.2 +/- 14.3; p < 0.05), while patients of group 2 (48.7 +/- 15.5) were older than those of group 4 (p < 0.01). More group 2 subjects had histories of jaundice (23/47) than those of group 4 (15/60; p < 0.05). Transaminase levels were higher in group 1 (median (range) IU/l of AST, ALT; 29 (13-95), 32 (9-144) and group 3 (25 (15-187), 22 (8-185)) than in group 4 (18 (9-13), 15 (6-133); p < 0.05, p < 0.005 vs. group 1, and p < 0.005, p < 0.001 vs. group 3, respectively). In HBsAg-negative subjects, those with higher titers of anti-HCV (cut-off index > 15) were older, and had more histories of jaundice and higher levels of AST and ALT than anti-HCV negative subjects (50.3 +/- 14.8 vs. 39.1 +/- 14.3, p < 0.01; 15/28 vs. 15/60, p < 0.01; 22.5 (12-127) vs. 18 (9-93), p < 0.05; 20.5 (7-362) vs. 15 (6-133), p < 0.05; respectively). In conclusion, this preliminary surveillance for hepatitis B and C viral markers showed that both hepatitis viruses are prevalent and may cause liver diseases in Mongolia. A nation-wide survey for these viruses should be urged and preventive measures should be taken to suppress the spread and development of liver diseases in this country.