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Biomedical subjects

M Iizuka

Publications and source records attributed to M Iizuka.

At least 55 records · Page 3Linked to original sources

Detection and serological relationships of cymbidium mosaic potexvirus isolates.

Twenty-two isolates of Cymbidium mosaic virus (CyMV) were isolated from 35 orchid plants suspected of being infected with CyMV. Among the three methods used for detecting CyMV, immunoelectron microscopy (IEM-1) was shown to be the most sensitive method, being able to detect the virus in 71.43% of suspected CyMV-infected plants while the electron microscopic method and the indexing plant method could detect 51.43 and 42.86%, respectively. Out of 12 symptomless plants investigated, 25% were found by IEM-1 method to be infected with the virus. Purified CyMV were flexuous rods having lengths between 470-490 nm. A few end-to-end aggregates were also observed and the 280 260 absorbance ratios were from 0.884 to 0.929. The yield of CyMV was 31.07 to 44.09 mg per kg of Datura leaves. Antibodies against purified CyMV D2 were produced in rabbits and hens. The antibody titers in the yolk and sera of hens indicated that 0.5 mg of virus per immunization efficiently generated an abundant supply of IgY in the yolk, however 1 mg of virus per immunization gave a stronger immune response in both sera and yolk. The average yields of IgY were 6.5 +/- 0.6 and 9.4 +/- 0.9 mg/ml of yolk in the group that received 0.5 mg and the group that received 1.0 mg of the virus, respectively. Positive ELISA reactions were observed in 18 and 20 of 22 CyMV isolates when detected with rabbit IgG and IgY, respectively, demonstrating that those isolates were serologically related and the ELISA reactions were shown to be stronger with IgY than those with rabbit IgG in most isolates. The degree of reaction between the CyMV isolates, O(2) and O(4), and IgY was less than that of the other isolates. The two isolates, D(6) and Cat(6), gave negative reactions to rabbit IgG. The results of ELISA assays showed that the homologous serological reaction was not consistently stronger than the heterologous one. Twelve isolates out of twenty-two gave stronger reactions than the homologous antigen (CyMV D(2)) when IgY was used as the detecting antibody while nine isolates gave stronger reactions when using rabbit IgG. No reactions were observed with other plant viruses and plant proteins from healthy Datura.

Journal Article↗

HLA-A33 and -B44 and susceptibility to postherpetic neuralgia (PHN).

HLA class I and class II alleles of 32 Japanese patients with postherpetic neuralgia (PHN) and 136 healthy controls were analyzed by serological (class I) and DNA (class II) typing for any significance in the susceptibility to varicella-zoster virus (VZV). We recognized positive associations of the development of PHN with the HLA class I antigens HLA-A33 and -B44, and the HLA-A33-B44 haplotype. This haplotype is tightly linked to DRB1*1302 in a Japanese healthy population. However, no significant association between PHN and HLA class II alleles was observed with no linkage of the HLA haplotype HLA-A33-B44 to HLA-DRB1*1302 in the patients with PHN. These findings suggest that HLA class I gene may genetically control the immune response against VZV in the pathogenesis of PHN.

Aged↗

Association analysis using refined microsatellite markers localizes a susceptibility locus for psoriasis vulgaris within a 111 kb segment telomeric to the HLA-C gene.

The HLA-Cw6 antigen has been associated with psoriasis vulgaris despite racial and ethnic differences. However, it remains unclear whether it is the HLA-Cw6 antigen itself or a closely linked, hitherto unidentified, locus that predisposes to the disease. Here, in order to map the susceptibility locus for psoriasis vulgaris precisely within the HLA class I region, 11 polymorphic microsatellite markers distributed throughout a 1060 kb segment surrounding the HLA-C locus were subjected to association analysis in Japanese psoriasis vulgaris patients. Statistical analyses of the distribution and deviation from Hardy-Weinberg equilibrium of the allelic frequency at each micro-satellite locus revealed that the pathogenic gene for psoriasis vulgaris is located within a reduced interval of 111 kb spanning 89-200 kb telomeric of the HLA-C gene. In addition to three known genes, POU5F1, TCF19 and S, this 111 kb fragment contains four new, expressed genes identified in the course of our genomic sequencing of the entire HLA class I region. Therefore, these seven genes are the potential candidates for susceptibility to psoriasis vulgaris.

Adult↗

Follow-up of clinical efficacy of iontophoresis therapy for postherpetic neuralgia (PHN).

A great variety of therapies have been attempted for PHN, including pharmacotherapy and physical therapy. However, there has been no decisive treatment, and reports of the clinical efficacy of all available therapies have been rather controversial. Almost all studies conducted so far have looked only at short-term therapeutic efficacy, and only a few investigators have conducted long-term observations or studies on long-term outcome. We followed up the clinical efficacy of iontophoresis therapy using lidocaine and methylprednisolone in 197 PHN patients. Monitoring conducted for an average of 4 years after completion of the treatment showed that pain remained unchanged or improved compared to pain observed upon completion of the treatment in 90.4% of patients. Although 42.6% of patients were still continuing some treatment, 90.9% were found to be able to take care of themselves. Findings obtained were reviewed and discussed from various viewpoints. Our findings showed that iontophoresis therapy is not only effective at the end of the treatment, but its efficacy is maintained over a long period of time, indicating that it is clinically very useful for the treatment of PHN.

Adult↗

Rostrocaudal progression in the development of periodic spontaneous activity in fetal rat spinal motor circuits in vitro.

Rostrocaudal progression in the development of periodic spontaneous activity in fetal rat spinal motor circuits in vitro. Developmental changes in the periodic spontaneous bursts in cervical and lumbar ventral roots (VRs) were investigated using isolated spinal cord preparations obtained from rat fetuses at embryonic days (E) 13.5-18. 5. Spontaneous bursts were observed in the cervical VR at E13.5-17.5, and in the lumbar VR at E14.5-17.5. Bursts occurrence in the cervical and lumbar VRs was correlated in a 1:1 fashion at E14.5-16. 5. The bursts in the cervical VR preceded those in the lumbar VR at E14.5, but the latter came to precede the former by E16.5. The interval between spontaneous bursts in the lumbar VR was greatly prolonged after spinal cord transection at the midthoracic level at E14.5, whereas that in the cervical VR became significantly longer at E14.5-16.5. These results suggest that the dominant neuronal circuit initiating the spontaneous bursts shifts from cervical to lumbar region during this period. Bath application of a glutamate receptor antagonist, kynurenate (4 mM), had little effect on the spontaneous bursts in either cervical or lumbar VRs at E14.5-15.5. At E16.5, kynurenate abolished the spontaneous bursts in the cervical VR. Concomitant application of kynurenate and strychnine (5 microM), a glycine receptor antagonist, abolished all spontaneous bursts, suggesting that the major transmitter mediating the spontaneous bursts changes from glycine to glutamate in the cervical region by E16.5, but not in the lumbar region during this period.

Animals↗

[Fundamental study of the detection of metastatic bone tumors with whole-body MR imaging].

Preliminary studies were performed to evaluate the usefulness of whole-body magnetic resonance imaging (WB-MRI) in the screening of bone metastases. The fast short TI inversion recovery sequence was the most suitable method because of its minimal image distortion and uniformity of fat suppression in a large field of view. In 4 cases of malignant tumor, the localization of all 8 bone metastases detected with bone scintigraphy (BS) could be clarified with WB-MRI. Furthermore, WB-MRI with fast STIR alone could depict 4 lesions undetected by BS, and WB-MRI combined with contrast enhancement could depict 5 such lesions.

Bone Neoplasms↗

[Granulomatous slack skin].

A 48-year-old woman developed granulomatous slack skin (GSS), one of the special forms of cutaneous T-cell lymphoma. The lesional skin slack with an atrophic, poikilodermic surface and granulomatous induration. Histopathological findings included epidermotropism, diffuse lymphoid cell infiltration and foreign body giant cells as well as granulomatous reactions from superficial to deep dermis, including part the subcutis. The diagnosis was established by positive results for rearrangement of the T-cell receptor gene. The therapeutic possibilities, especially with corticosteroids and monitoring the disease course by following serum angiotensin converting enzyme activity are discussed.

Breast↗

No Mycobacterium paratuberculosis detected in intestinal tissue, including Peyer's patches and lymph follicles, of Crohn's disease.

To clarify the etiologic significance of Mycobacterium paratuberculosis in Crohn's disease, we investigated whether M. paratuberculosis was detected in intestinal tissues, including Peyer's patches, where M. paratuberculosis invades, and colonic lymph follicles, where early lesions appear. Fifty-one samples of intestinal tissues, either therapeutically resected or biopsied, including 34 specimens from 30 patients with Crohn's disease, were studied. Four Peyer's patches and eight lymph follicles were included in the intestinal tissue samples of Crohn's disease. They were visualized by acetic acid fixation. DNA extracted from intestinal tissues by proteinase K treatment was used for nested polymerase chain reaction (PCR) for detection of IS900, which is specific for M. paratuberculosis. PCR products were analyzed by agarose gel electrophoresis and subsequent Southern blot analysis. Our amplification system could detect 7.5 fg of M. paratuberculosis DNA. None of the tissue samples showed positive IS900 amplification, whereas they all showed amplification of the positive control human leukocyte antigen (HLA)-DQA DNA. Spiked experiments of tissue samples with M. paratuberculosis demonstrated that inhibitors of IS900 amplification were not present in the samples. Our study does not support the etiologic significance of M. paratuberculosis in Crohn's disease.

Adolescent↗

Development of the spatial pattern of 5-HT-induced locomotor rhythm in the lumbar spinal cord of rat fetuses in vitro.

Developmental changes in the 5-hydroxytryptamine (5-HT)-induced locomotor rhythm were examined in isolated spinal cord preparations taken from fetal rats at embryonic day (E) 16.5, E18.5 and E20.5. Motor activity was recorded from L2/L3 and L5 ventral roots. Bath application of 5-HT evoked rhythmic bursts that were synchronized in all ventral roots studied at E16.5. At E18.5, 5-HT evoked rhythmic bursts that alternated between the two sides, while the bursts in the L2/L3 ventral root were synchronous with those in the ipsilateral L5 ventral root. At E20.5, 5-HT evoked alternate rhythmic bursts in L2/L3 and L5 ventral roots, representing activity in flexors and extensors, respectively. In the presence of strychnine, 5-HT induced rhythmic bursts that were synchronized in all ventral roots studied at E18.5 and E20.5, suggesting that the change in the pattern of rhythmic motor activity that occurs with age is due to the development of glycine-mediated inhibition.

Animals↗

Microbiology of the intestinal lymph follicle: a clue to elucidate causative microbial agent(s) in Crohn's disease.

It has been suggested that microbial agent(s) are involved in the onset of Crohn's disease. None of the candidates, however, has been unequivocally demonstrated to be a causative agent. The macroscopically earliest lesion takes place in the lymph follicle, irrespective of the initial attack or relapse in Crohn's disease. Human leucocyte antigen-DR (HLA-DR) antigens are expressed on the epithelium around the lymph follicle even in areas endoscopically uninvolved in Crohn's disease. These observations make the lymph follicle critical in the onset of Crohn's disease. The lymph follicle is a port of entry of a variety of microbial agent(s), leading to the speculation that microbial agent(s) exist in the lymph follicle. Polymerase chain reaction (PCR) using universal primers designed from conserved regions of bacterial ribosomal RNA or techniques such as representational difference analysis, may well identify microbial agent(s) in the lymph follicle that are specific to Crohn's disease. The existence of bacteria in the lymph follicle is here indicated by preliminary studies.

Crohn Disease↗

New method for determining prognosis of patients with psoriasis (E-PAP).

We developed a new method for evaluating clinical symptoms of psoriasis during the observation period, by adding a parameter of time (the number of days) to the psoriasis area and severity index (PASI). This method was named the evaluation for prognosis with averaged PASI (E-PAP). Using this method, we assessed 14 cases to determine the following items: (1) clinical symptoms during the observation period; (2) clinical effects of treatments; (3) patient's satisfaction with treatments; and (4) necessity for additional and/or alternate treatments. We evaluated the usefulness of E-PAP in determining the prognosis of patients with psoriasis by comparing the uninterrupted monotherapy and intermittent therapy with cyclosporin A (Cys A). Although there were no differences in Cys A dosages after remission as well as adverse reactions between the two groups, the E-PAP value was significantly different between them. These results suggest that this method may be useful for determining the prognosis of patients with psoriasis.

Antirheumatic Agents↗

Histological and immunocytochemical studies of human psoriatic lesions transplanted onto SCID mice.

To investigate the pathology of psoriasis, we developed an animal model for this disease using severe combined immunodeficiency (SCID) mice. These mice possess neither B nor T Lymphocytes so that both cellular and humoral immunities are impaired. For the in vivo study of psoriasis, human psoriatic skin was grafted on SCID mice. Long-term morphological and immunohistochemical changes in the grafted skin ware examined for up to 22 weeks after transplantation. The human skin graft were generally well maintained during this period, but the histological and immunohistochemical findings characteristic of psoriasis, except for acanthosis and hyperkeratosis, gradually disappeared as lymphocytic infiltration of the psoriatic lesions declined.

Adult↗

Interaction of cultured keratinocytes and fibroblasts from human psoriatic and normal skin in immunodeficient mice.

Clarification of the pathogenesis of psoriasis requires separate studies of the epidermis, dermis, and inflammatory cells. We previously subcutaneously transplanted a mixture of cultured human keratinocytes and fibroblasts into mice to develop cysts with human skin structures. Using this method, we separately cultured psoriatic and normal keratinocytes and fibroblasts. Four mixtures were prepared: normal keratinocytes and normal fibroblasts (NK/NF); psoriatic keratinocytes and normal fibroblasts (PK/NF); normal keratinocytes and psoriatic fibroblasts (NK/PF); and psoriatic keratinocytes and psoriatic fibroblasts (PK/PF). Each mixture was transplanted into immunodeficient mice to observe formation of cysts and histological changes. The cysts varied in structure depending on the mixture, which suggests that psoriatic keratinocytes and fibroblasts had some abnormalities. Psoriatic fibroblasts may be partially responsible for thickening of the epidermis. Cell differentiation might have been accelerated in psoriatic keratinocytes after transplantation, resulting in the loss of epidermis structures.

Animals↗

Listeria monocytogenes in Crohn's disease.

BACKGROUND: In an immunohistochemical study a higher rate of reactivity of intestinal tissues to the antibody against Listeria monocytogenes was reported in Crohn's disease as compared with controls. METHODS: Seventy-six intestinal tissues, either therapeutically resected or biopsied, from 31 patients with Crohn's disease, 20 with ulcerative colitis, and 21 with non-inflammatory bowel disease were studied. DNA extracted from intestinal tissues by proteinase K treatment was used for nested polymerase chain reaction (PCR), using two sets of primers. PCR products were analyzed with agarose gel electrophoresis and subsequent Southern blot analysis. RESULTS: Our amplification system could detect 9 pg of L. monocytogenes DNA. L. monocytogenes was detected in only one sample, that from a patient with ulcerative colitis. CONCLUSIONS: Our study does not support the etiologic significance of L. monocytogenes in Crohn's disease.

Adult↗

Cloning and sequencing of cDNA encoding ribonuclease F1 from Fusarium moniliforme.

Two cDNA clones encoding ribonuclease F1 (EC 3.1.27.3) have been isolated using a probe prepared by polymerase chain reaction with primers designed on the basis of amino acid sequence of the enzyme. They derived probably from the same gene and contained 393-base pair open reading frame encoding 131 amino acid residues (Mr 13,606) including a putative 25-residue signal peptide. The sequences of 43-base pair 5'-untranslated region and 125-base pair 3'-untranslated region including a poly(A) tail of 25 nucleotides were also elucidated. Homology analyses showed that cDNA for ribonuclease F1 has 65% homology to that for ribonuclease T1 in the coding region. At the preprotein level, they share 53% homology.

Amino Acid Sequence↗

Novel vaginal danazol ring therapy for pelvic endometriosis, in particular deeply infiltrating endometriosis.

Danazol is routinely administered orally to inhibit ovulation and to treat pelvic endometriosis. However, recent evidence suggests that danazol can act directly on endometriotic tissue in vitro to inhibit DNA synthesis and induce apoptosis. Danazol was administered via the vagina in this study, using a vaginal ring drug delivery system containing 1500 mg of danazol. This therapy was effective for treatment of pelvic endometriosis, especially for deeply infiltrating endometriosis, resulting in a cure of dysmenorrhoea and tenderness in the cul-de-sac within 3 months, and of induration or nodularity in the cul-de-sac within 7 months. Moreover, conception was possible during insertion of the vaginal ring in 17 out of 31 infertile women with deeply infiltrating endometriosis, and in two out of eight infertile women with ovarian endometriotic cysts not adhering to the cul-de-sac and without deeply infiltrating endometriosis. Serum danazol concentrations, high during oral daily 400 mg danazol therapy, but undetectable during vaginal danazol ring therapy, explain why ovulation and conception could occur during insertion of the vaginal danazol ring, and why general side-effects, which are often observed during oral danazol therapy, were not observed during vaginal danazol ring therapy. Danazol seems to be absorbed through the vaginal mucosa and reaches the deeply infiltrating endometriosis via diffusion.

Absorption↗

HLA class I and II alleles and susceptibility to generalized pustular psoriasis: significant associations with HLA-Cw1 and HLA-DQB1*0303.

HLA alleles in generalized pustular psoriasis (GPP) were investigated to clarify the etiology and/or pathogenesis of this disease. Not only serological typing of HLA class I and II antigens but also genotyping of HLA class II alleles were carried out in twenty-six unrelated Japanese patients with GPP. These patients were classified according to their history of psoriasis vulgaris (PV). Serological typing revealed a significantly high incidence of HLA-Cw1 (Pc = 0.04) in the patients as compared with Japanese healthy controls. The frequency of HLA-B46 was particularly high in the patients with GPP and a previous history of PV. Genotyping of HLA class II alleles showed a highly significant increase in HLA-DQB1*0303 (Pc = 0.01) in the patients vs. the healthy controls. In particular, HLA-DQB1*0303 was significantly more frequent in the patients with no prior history of PV than in those with a history of PV. Analysis on linkage disequilibrium showed remarkably different patterns for HLA class II haplotypes between the patients and the healthy controls. Based on the comparative analysis among the amino acid sequences of the beta 1-domain of the HLA-DQB1*03 alleles, proline at residue 55 was suggested to be important as a common amino acid for determination of the susceptibility to GPP. These results revealed not only an association between the etiology and/or pathogenesis of GPP and HLA, but also different mechanisms of the immune response between the patients with GPP and PV.

Amino Acid Sequence↗