Biomedical subjects

M Fukayama

Publications and source records attributed to M Fukayama.

At least 109 records · Page 6Linked to original sources

PubMed1993

Systematic analysis of glycosphingolipids in the human gastrointestinal tract: enrichment of sulfatides with hydroxylated longer-chain fatty acids in the gastric and duodenal mucosa.

The composition of the glycosphingolipids of the human gastrointestinal tract was studied. The major neutral glycosphingolipids were ceramide monohexosides (e.g., GalCer, GlcCer), LacCer, Gb3Cer, Gb4Cer and more polar ones with more than four sugars, whereas neither Gg3Cer nor Gg4Cer were present. The acidic glycosphingolipids consisted of sulfatides and gangliosides such as GM3, GM1, GD3 and GD1a. Also a large amount of sulfatides was found in the gastric mucosa and duodenum. The concentrations of sulfatides in the fundic mucosa, antral mucosa and duodenum amounted to 416.0, 933.8 and 682.9 nmol/g of dry weight, respectively, exceeding those in the gastric mucosa and kidney of other mammals. The major molecular species of the sulfatides were identified as I3SO3-GalCer with hydroxylated longer-chain fatty acids based on the analyses by gas-liquid chromatography and negative ion fast-atom bombardment mass spectrometry. In contrast, gangliosides in these regions showed a tendency to be lower than sulfatides, and the molar ratios of sulfatides to gangliosides were about 2.0, whereas those in other parts were less than 0.5. A high content of sulfatides in the gastric and duodenal mucosa, where mucosa is easily insulted by acid, pepsin and bile salts, may be closely related to their roles in mucosal protection.

Carbohydrates↗

PubMed1993

In vivo stimulation of thyroid hormone release by rabbit antibodies with thyroid stimulating activity raised against the mutated receptor peptide TSH-R-HIS17.

The present study was designed to examine whether thyroid stimulating antibodies (TSAb) against the mutated or non-mutated peptide of human TSH receptor stimulates release of thyroid hormones in mice in vivo. We generated antibodies against mutated TSH receptor peptide TSH-R-HIS17 and non-mutated TSH receptor peptide TSH-R-ASP17 immunizing the peptides to rabbits. We isolated immunoglobulin G (IgG) from these antibodies and injected iv into male ddy mice. Control mice were injected iv with bTSH or TSAb-negative normal rabbit IgG. Serum T3 and T4 concentrations were elevated after the administration of bTSH in a dose-dependent manner. Serum T3 and T4 concentrations were significantly elevated after the injection of IgG from the rabbit antibodies with TSAb activity produced against TSH-R-HIS17. However serum T3 and T4 remained within normal levels after the injection of IgG from the rabbit antibodies without TSAb activity toward TSH-R-ASP17. The present data offer in vivo support of the IgG with TSAb activity interaction with the TSH receptor of the thyroid and raise the possibility that TSAb against mutated human TSH receptor peptide might be contributed to induce thyroid hyperfunction.

Amino Acid Sequence↗

PubMed1993

Cell-cycle analysis detecting endogenous nuclear antigens: comparison with BrdU-in vivo labeling and an application to lung tumors.

The versatility of non-radioactive cell-cycle analysis in detecting endogenous nuclear antigens of the proliferating cells was evaluated. Optimal conditions for immunostaining varied in fixation and pretreatment procedures among antigens, bromodeoxyuridine (BrdU), Ki-67 epitope, DNA polymerase alpha and PCNA. A significant correlation between BrdU labeling index (LI) was observed in each positive ratio (PR, positive/total neoplastic cells) for nuclear antigens in tumor-sections which had been labeled in vivo with BrdU. The best correlation was observed in Ki-67 PR (y = 1.26x + 2.5; y = Ki-67 PR; x = BrdU LI; r = 0.97). To determine its prognostic value, Ki-67 analysis was applied to the surgically resected lung tumors. Ki-67 PR were different according to the histologic types of the tumors: 47.8 +/- 3.4% in small cell carcinoma; 29.5 +/- 3.5% in squamous cell carcinoma; 28.3 +/- 4.7% in large cell carcinoma; 15.2 +/- 1.8% in adenocarcinoma and 0.1 +/- 0.1% in mature carcinoid tumor. When the mean value was used to divide each type to a higher or lower proliferative activity (15% Ki-67 PR for adenocarcinoma and 30% for squamous cell carcinoma), the group with the lower Ki-67 PR showed a significantly more favorable prognosis than that of a higher ratio. Ki-67 PR was not correlated with other pathologic factors such as size, lymph node metastasis or pleural involvement. Non-radioactive cell-cycle analysis was feasible and useful for detecting endogenous nuclear antigens even in the lung tumors, particularly when the analysis was coupled with histologic typing.

Antibodies, Monoclonal↗

PubMed1993

Production of bioactive rabbit antibodies against the mutated receptor peptide TSH-R-HIS17 and detection of hypertriiodothyroninemia in the rabbit.

We produced rabbit antibodies against mutated thyrotropin (TSH) receptor peptide TSH-R-HIS17 in comparison with non-mutated TSH receptor peptide TSH-R-ASP17. All the antibodies raised against TSH-R-HIS17 showed thyroid stimulating antibody (TSAb), thyroid stimulation blocking antibody (TSBAb) and/or TSH binding inhibiting immunoglobulin (TBII) activities and also those serum T3 levels were all in thyrotoxic levels associated with TSAb activities. However, all the antibodies raised against TSH-R-ASP17 had no activities of TSAb, TSBAb and/or TBII and those serum T3 levels were all within normal range. The data suggest that the mutation in the TSH receptor gene may have a possible relevance for the pathogenesis of Graves' disease.

Amino Acid Sequence↗

PubMed1993

Histological studies on the effects of tooth brushing on repair of alveolar bone after periodontal osseous surgery in the rat incisor.

The purpose of this study was to elucidate the effects of tooth brushing on repair of alveolar bone after periodontal osseous surgery in the labial alveolar bone of rat incisor. The surgery was performed on 24 Wistar rats divided into 2 groups: the experimental group, which was subjected to tooth brushing, and the control group, which was not. In the experimental group, daily tooth brushing was initiated at 4 weeks after surgery. The rats were sacrificed after 1 or 2 weeks of tooth brushing. Microradiographic, light and fluorescence microscopic examinations were made of sections of the alveolar bone and its surrounding tissue. After 1 week of tooth brushing, callus with a low degree of mineralization and with large, irregularly arranged, young osteocytes appeared in the superficial layer and crest portion of alveolar bone in the brushing region. Numerous blood vessels had invaded the callus. In this region, the height of osteoblasts on the callus surface increased. At the alveolar crest region, the callus was approximately 3 times thicker than in the superficial region. After 2 weeks of tooth brushing, modification had occurred in the callus; this region had evolved into developed bone with a compact matrix. These findings suggest that the intermittent mechanical stress of tooth brushing is useful in activating the cells of the alveolar periosteum and in stimulating bone formation.

Alveolar Process↗

PubMed1993

Epstein-Barr virus in pyothorax-associated pleural lymphoma.

Pleural B-cell lymphoma was found in five patients with a history of pyothorax that was the sequelae of tuberculosis 35 to 47 years previously. Epstein-Barr virus (EBV) DNA was detected in all five pleural tumors by polymerase chain reaction and Southern blot hybridization. The lymphoma cells were shown to express the latent membrane protein-1 and the EBV-encoded nuclear antigen-2 by immunocytochemistry and EBV-encoded small RNA by in situ hybridization. Three cases were shown to be EBV subtype A, whereas the remaining two were subtype B, as determined by differences in the EBV-encoded nuclear antigen-2 nucleotide sequence. The patients also had high titers of antibodies against EBV. These findings suggest that EBV is causally associated with the pleural lymphomas that originate at the site of chronic inflammation and fibrosis with a latent period of more than 40 years.

Adult↗

PubMed1992

Genetic changes of both p53 alleles associated with the conversion from colorectal adenoma to early carcinoma in familial adenomatous polyposis and non-familial adenomatous polyposis patients.

Mutation and loss of heterozygosity (LOH) in the p53 gene were analyzed in 274 colorectal tumors of 4 histopathological grades. Among 160 tumors from 40 familial adenomatous polyposis patients, none of 58 adenomas with moderate dysplasia had p53 mutations, whereas 8% (3 of 37) of severe adenomas, 15% (6 of 40) of intramucosal carcinomas, and 40% (10 of 25) of invasive carcinomas had p53 mutations. Only 3% (1 of 33) of severe adenomas showed both mutation and LOH, while 25% (6 of 24) of intramucosal carcinomas and 40% (10 of 25) of invasive carcinomas had both mutation and LOH. All intramucosal and invasive carcinomas that had mutations lost the other allele of the p53 gene. In 114 tumors from 86 non-familial adenomatous polyposis patients, similar results were obtained; no adenoma showed both mutation and LOH, but both alterations occurred in intramucosal and invasive carcinoma. As regards specificity in 56 mutations detected in the present study, the frequently affected codons were codons 175, 238, 245, 248, 273, and 282, 4 of these amino acids being arginine, and 72% (39 of 54) of all mutations were GC to AT transition. Although expression into p53 polyadenylated RNA was high in every invasive carcinoma irrespective of the presence of mutation or LOH, there was a correlation between mutation and protein level; immunostaining of p53 protein was negative in almost all adenomas, but it was positive in 86% of invasive carcinomas exhibiting p53 mutation. These data suggest that genetic changes on both alleles of the p53 gene through mutation and LOH, which result in abnormal protein accumulation, are involved in the conversion of adenoma to early carcinoma. Also, carcinoma cells with p53 mutations existing within adenoma tissues are detectable by immunostaining, even in formalin-fixed, paraffin-embedded specimens.

Adenoma↗

PubMed1992

Differential age-change in the numbers of CD4+CD45RA+ and CD4+CD29+ T cell subsets in human peripheral blood.

Peripheral blood mononuclear cells were obtained from people ranging in age from newborn to 102 years old and analyzed by dual color flow cytometer in terms of number and percentage of various subsets of T cells, B cells and natural killer cells (CD3, 4, 5, 8, 11b, 19, 20, 21, 25, 29, 45RA and 56). Numbers of T cells (CD3+ or CD5+ cells) significantly declined at the 3rd decade as compared with those of younger people, stayed at a relatively constant level between the 3rd and the 7th decade and gradually declined thereafter. In T cell subsets, both CD4 and CD8 positive positive cells decreased with age, but a decrease was more pronounced in the latter, showing an age-related increase of CD4/CD8 ratio. The most interesting finding was a contrasting age-change in two subsets of CD4+ T cells; i.e. a subset of suppressor inducer T cells (CD4+CD45RA+ naive cells) decreased with age, while a subset of helper inducer T cells (CD4+CD29+ memory cells) increased with age. CD20+ B cells also decreased with age in a manner similar to that observed in T cells. Natural killer cells (CD56) showed an increase in numbers with age. The relationship between these changes in various subsets of peripheral blood leukocytes and the age-related decline in immune functions has been discussed.

Adolescent↗

PubMed1992

In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium.

Direct transfer of the normal cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene to airway epithelium was evaluated using a replication-deficient recombinant adenovirus (Ad) vector containing normal human CFTR cDNA (Ad-CFTR). In vitro Ad-CFTR-infected CFPAC-1 CF epithelial cells expressed human CFTR mRNA and protein and demonstrated correction of defective cAMP-mediated Cl- permeability. Two days after in vivo intratracheal introduction of Ad-CFTR in cotton rats, in situ analysis demonstrated human CFTR gene expression in lung epithelium. PCR amplification of reverse transcribed lung RNA demonstrated human CFTR transcripts derived from Ad-CFTR, and Northern analysis of lung RNA revealed human CFTR transcripts for up to 6 weeks. Human CFTR protein was detected in epithelial cells using anti-human CFTR antibody 11-14 days after infection. While the safety and effectiveness remain to be demonstrated, these observations suggest the feasibility of in vivo CFTR gene transfer as therapy for the pulmonary manifestations of CF.

Adenoviruses, Human↗

PubMed1992

Expression of the C3d-binding protein (CR2) from Candida albicans during experimental candidiasis as measured by lymphoblastogenesis.

The complement C3d-binding protein (CR2) of Candida albicans has been purified by immunoaffinity chromatography, and its specificity has been characterized by immunoblotting with monoclonal antibodies to the C. albicans CR2 and the mammalian CR2. Recent studies with immunoelectron microscopy indicated that the CR2 was expressed during a systemic infection in a murine model of candidiasis. As a continuation of these observations, the immunogenicity of the C. albicans CR2 was investigated in a lymphoblastogenesis assay. Lymph node cells as well as splenic lymphocytes from mice infected subcutaneously with viable blastoconidia of C. albicans reacted to the C. albicans CR2 to a significantly greater extent than did lymphocytes from uninfected mice (P less than 0.01). The maximum stimulation of splenic lymphocytes by the purified receptor occurred at a concentration of 0.54 micrograms of protein per ml after 72 h of incubation of lymphocytes and receptor. Also, splenocytes from infected or CR2-immunized mice exhibited significantly reduced responses to the T-cell-dependent mitogen phytohemagglutinin (P less than 0.01). These data indicate that lymphocytes from infected mice respond to the C. albicans CR2 in a lymphoproliferation assay to a greater extent than do lymphocytes from uninfected mice, indicating that the CR2 is expressed in vivo.

Animals↗

PubMed1992

[A case report of severe urinary retention and meteorism during flunarizine administration].

A case of flunarizine hydrochloride (FZ)-induced severe urinary retention and meteorism which resulted from sphincter spasm of the urinary bladder and the anus is presented. An 81-year-old female had received 10 mg/day FZ orally for 12 months before hypokinesia and general fatigue developed. Physical examination revealed slight rigidity of the extremities, abdominal distention and spasm of the anal sphincter muscle. Laboratory examinations showed uremia (BUN 88 mg/dl, Creatinine 16.8 mg/dl) and abdominal X-ray demonstrated marked distention of the small and large bowels. Renal failure improved within 2 days after massive urination using a urethral catheter. Abdominal distention was improved by the ileus and anal tubes. The difficulties of urination and defecation and decreased mobility of the extremities were resolved one month after the cessation of FZ. No organic changes were detected in urinary, intestinal and neurological systems by cystoscopy, CT, MRI and gastrointestinal fiberscopy. Serum concentration of FZ was 42.5 ng/ml on admission but decreased slowly to 17.9 ng/ml 80 days later. Serum half life was calculated to be 55 days which was 3 times longer than that healthy younger volunteers.

Acute Kidney Injury↗

PubMed1992

[Clinical spectrum of antibiotic associated enterocolitis due to methicillin resistant Staphylococcus aureus].

Historically, various types of antibiotic-associated diarrhea have been recognized. In the 1950-1960s, multi-resistant Staphylococcus aureus was implicated to be major cause of antibiotic-associated diarrhea, especially pseudomembranous enterocolitis. In the late 1970s, a new type of antibiotic-associated diarrhea, which is characterized by pseudomembranous colitis and lack of fecal multi-resistant S. aureus, usually following lincosamide administration, was recognized. Many studies have revealed the pathogenic roles of Clostridium difficile in these patients. Recent emergence of methicillin resistant S. aureus (MRSA) provokes another type of antibiotic-associated diarrhea. We have tried to isolate S. aureus and C. difficile from 150 faecal samples of patients with antibiotic-associated diarrhea. S. aureus alone was isolated from 32 (21.3%) samples. C. difficile alone was isolated from 51 (34.0%) samples, and, both were detected in 23 (15.3%) samples. 90.9% of S. aureus were MRSA. Comparing the clinical features of these cases, patients with both C. difficile and MRSA showed more serious clinical features. The patients who had undergone laparotomy were accompanied by shock and meteorism, more often than patients without laparotomy. The management of antibiotic-associated diarrhea due to MRSA is discussed.

Aged↗

PubMed1992

[Methicillin resistant Staphylococcus aureus (MRSA) infection in geriatric hospital--management for infected decubital ulcer].

Wide spread of MRSA infection in geriatric hospitals indicated two important problems which must be resolved. One is how we can eradicate MRSA in the elderly, and the other is how we can control cross infections of MRSA within the hospital. The presence of infected decubital ulcers in the elderly make it difficult to answer these problems. Usually, MRSA and Pseudomonas aeruginosa which is isolated from decubital ulcers act as colonizers on the surface of granulated wounds but do not contribute to clinical symptoms. However, these colonizers can be the source of cross infection toward other debilitated patients. The actual management for decubital ulcer infected by MRSA are discussed from the above viewpoints.

Aged↗

PubMed1992

[A case of coexistent bronchogenic carcinoma with pulmonary tuberculosis].

A 79-year-old housewife complaining of shortness of breath presented with a LUL mass lesion on chest roentgenogram. Bronchofiberscopic biopsy and curettage revealed adenocarcinoma as well as acid-fast bacilli of Gaffky-4. Further investigations showed increased ESR, CRP levels and positive RF. PPD skin test was positive measuring 55 x 43 mm. A left upper lobectomy with mediastinal lymph node dissection was performed. The mass was in S1+2 and measured 2.7 x 2.5 x 1.5 cm. Histologically caseaous necroses and spotted granulomas of tuberculosis were surrounded by bronchioloalveolar cell carcinoma. Recent discussion concerning the simultaneous occurrence of pulmonary tuberculosis and bronchogenic carcinoma suggests a higher coexistence of both diseases. However, the coexistence of active tuberculosis with carcinoma in the same region, as in our case, is quite rare and suggests an etiological closer relationship between both diseases.

Adenocarcinoma↗

PubMed1992

[Pulmonary lesions of acquired immunodeficiency syndrome--analysis of 24 Japanese autopsy cases with AIDS].

The pulmonary lesions were studied in 24 autopsy cases of Japanese patients with AIDS. The major pathological findings were opportunistic infections, which were the major clinical symptoms in some patients. The pathogens identified were as follows; Pneumocystis carinii (PC) in 10, cytomegalovirus (CMV) in 14, atypical mycobacterium in 5, cryptococcus in 2, candida in 2, and nocardia in 1. PC pneumonia was prominent in 8 cases and was the cause of death. In such patients, the lung were heavy and appeared parenchymatous. Histological examination revealed numerous protozoa in the foamy material in the alveolar spaces, associated with swelling of the alveolar lining cells and edematous thickening of the alveolar septa. In some cases, only hyaline membrane formation was prominent without foamy material in the alveolar spaces. Immunostaining with anti-PC monoclonal antibody or in-situ hybridization with oligopeptide demonstrated pathogens in the hyaline membranes. Many cases with PC pneumonia had concomitant opportunistic infections such as CMV, Herpes simplex virus, and atypical mycobacterium. Extrapulmonary infection of PC was seen in only one case. CMV infection was found in 14 cases; 7 had innumerable inclusion bodies, and in some cases the lesions were most prominent around the bronchioles. Of the 5 cases of atypical mycobacterial infection, 2 were caused by M. kansaii (MK) and 3 by M. avium intracellulare (MAI). Both lesions of MK infection showed necrosis and cavitation. One of three cases of MAI infection showed cavitation. Around the cavitary lesions, numerous cytomegalic inclusion bodies were identified in the mesenchymal cells, which may have been the cause of necrosis and cavitation of the lesions. MAI infection was systemic and pronounced in the lymph nodes, spleen, and intestinal mucosa. Neoplastic lesions comprised 2 cases of Kaposi's sarcoma and 4 of extranodal non-Hodgkin lymphoma in other organs. Lung involvement was seen in only one case of Kaposi's sarcoma although very small in size. The lesion was situated along the pulmonary vein and appeared hemorrhagic macroscopically. Pulmonary lesions in AIDS are complicated, and many of opportunistic pathogens were identified in single patients.

Acquired Immunodeficiency Syndrome↗

PubMed1991

Respiratory tract gene transfer. Transplantation of genetically modified T-lymphocytes directly to the respiratory epithelial surface.

To evaluate the strategy for potentially treating respiratory disorders with genetically modified T-lymphocytes, the interleukin-2 (IL-2)-dependent murine T-cell line, CTLL2, was genetically altered with the Escherichia coli beta-galactosidase (beta-gal) gene (lacZ) in vitro with a retroviral vector and the modified T-cells were transplanted directly to the respiratory epithelial surface of syngeneic C57Bl/6 mice. Southern and Northern analyses confirmed that the neomycin-selected modified T-cells contained and expressed the lacZ gene. The fate of the modified T-cells (CTLL2/lacZ) was followed by flow cytometry with T-cell surface marker Thy1.2 and fluorescent beta-gal analysis. One day after transplantation (7.5 x 10(5) CTLL2/lacZ T-cells/g of body weight), 95 +/- 3% of the Thy1.2+ T-cells recovered from respiratory epithelial lining fluid (ELF) were beta-gal+. Importantly, the modified T-cells remained in the lung for some time; at 3 days, Thy1.2+ beta-gal+ T-cells represented 63 +/- 12% of ELF Thy1.2+ T-cells and 59 +/- 6% of Thy1.2+ T-cells recovered from the whole lung. At 7 days, 33 +/- 8% of the Thy 1.2+ cells in ELF and 75 +/- 6% of the Thy1.2+ cells in whole lung were Thy1.2+ beta-gal+. In contrast, the proportion of the Thy1.2+ beta-gal+ T-cells in the spleen, the major extrapulmonary lymphatic organ, never rose above 3 +/- 1% of the total Thy1.2+ cells. The number of Thy1.2+ beta-gal+ T-cells in the lung could be modified by the systemic administration of IL-2, with whole lung Thy1.2+ beta-gal+ T-cells increasing 4.6-fold 3 days after transplantation, compared with non-IL-2-treated animals. These studies suggest that direct transplantation of genetically modified T-cells into the lung is feasible and represents a viable strategy for lung-specific gene transfer.

Animals↗

PubMed1991

Granulocyte-macrophage colony-stimulating factor promotes the proliferation of human alveolar macrophages in vitro.

The effects of granulocyte-macrophage (GM-CSF) or macrophage-CSF on in vitro proliferation of human alveolar macrophages (AM) were evaluated. AM of healthy volunteers incubated with recombinant human GM-CSF revealed incorporation of [3H]thymidine in vitro. The maximum incorporation was observed at 20 U/ml of GM-CSF on day 3. The proportion of proliferating cells incubated with 20 U/ml of GM-CSF from day 3 to day 4 was 8 to 11% of the total, whereas 3 to 5% of cells proliferated without GM-CSF. The number of cell nuclei increased 1.30- to 1.68-fold in the initial 7 days during incubation with 20 U/ml of GM-CSF, whereas there was a 1.07- to 1.13-fold increase without GM-CSF. Conditioned media obtained by the incubation with human lung tissue exhibited similar effects as recombinant human GM-CSF on macrophages. The effects were completely abrogated by antibody against human GM-CSF. Immunohistochemically, GM-CSF was detected in lung cells including AM, alveolar epithelium, alveolar interstitial cells, and endothelial cells. In contrast, recombinant macrophage-CSF did not induce the proliferation of human AM, although it has been known to promote the proliferation of murine AM. These observations suggest that GM-CSF plays an important role among the regulatory factors that locally support the population of AM in human lungs.

Adult↗

PubMed1991

Adenovirus-mediated transfer of a recombinant alpha 1-antitrypsin gene to the lung epithelium in vivo.

The respiratory epithelium is a potential site for somatic gene therapy for the common hereditary disorders alpha 1-antitrypsin (alpha 1AT) deficiency and cystic fibrosis. A replication-deficient adenoviral vector (Ad-alpha 1AT) containing an adenovirus major late promoter and a recombinant human alpha 1AT gene was used to infect epithelial cells of the cotton rat respiratory tract in vitro and in vivo. Freshly isolated tracheobronchial epithelial cells infected with Ad-alpha 1AT contained human alpha 1AT messenger RNA transcripts and synthesized and secreted human alpha 1AT. After in vivo intratracheal administration of Ad-alpha 1AT to these rats, human alpha 1AT messenger RNA was observed in the respiratory epithelium, human alpha 1AT was synthesized and secreted by lung tissue, and human alpha 1AT was detected in the epithelial lining fluid for at least 1 week.

Adenoviridae↗