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Biomedical subjects

K Kameda

Publications and source records attributed to K Kameda.

At least 73 records · Page 4Linked to original sources

Isolation and partial characterization of the gene for goose fatty acid synthase.

Fatty acid synthase is regulated by diet and hormones, with regulation being primarily transcriptional. In chick embryo hepatocytes in culture, triiodothyronine stimulates accumulation of enzyme and transcription of the gene. Since the 5'-flanking region of this gene is likely involved in hormonal regulation of its expression, we have isolated and partially characterized an avian fatty acid synthase gene. A genomic DNA library was constructed in a cosmid vector and screened with cDNA clones that contained sequence complementary to the 3' end of goose fatty acid synthase mRNA. A genomic clone (approximately 35 kilobase pairs (kb] was isolated, and a 6.5-kb EcoRI fragment thereof contained DNA complementary to the 3' noncoding region of fatty acid synthase mRNA. Additional cosmid libraries were screened with 5' fragments of previously isolated genomic clones, resulting in the isolation of five overlapping cosmid DNAs. The entire region of cloned DNA spans approximately 105 kb. Exon-containing fragments were identified by hybridization with end-labeled poly(A)+ RNA and by hybridization of labeled exon-containing genomic DNA fragments to fatty acid synthase mRNA. A new set of cDNA clones spanning approximately 3.2 kb was isolated from a lambda-ZAP goose liver cDNA library using the 5'-most exon-containing fragment of the 5'-most genomic DNA clone. This region of mRNA contains a 5'-untranslated sequence and a continuous open reading frame which includes a region that codes for the essential cysteine of the beta-ketoacyl synthase domain. The entire fatty acid synthase gene spans about 50 kb. The 5' 15 kb of the gene contain 7 exons. S1 nuclease and primer extension analyses were used to identify a single site for initiation of transcription, 174 nucleotides upstream from the putative translation initiation codon. Putative "TATA" and "CCAAT" boxes are located 28 and 60 base pairs (bp), respectively, upstream of the site of initiation of transcription. The 5'-flanking 597 bp of DNA contains G/C-rich sequences including several "GC" boxes corresponding to binding sites for the nuclear transcription factor Sp1. Putative sites for AP-2, C/EBP, and the triiodothyronine and glucocorticoid receptors also were found in this region. A chimeric DNA, containing approximately 1.6 kb of 5'-flanking sequence and 139 bp of untranslated sequence of the goose fatty acid synthase gene ligated to the bacterial chloramphenicol acetyl-transferase (CAT) gene, was transfected into chick embryo hepatocytes in culture. Cells treated with triiodothyronine contained increased chloramphenicol acetyltransferase and fatty acid synthase activities.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Sperm immobilizing and fertilization-blocking monoclonal antibody 2C6 to human seminal plasma antigen and characterization of the antigen epitope corresponding to the monoclonal antibody.

A monoclonal antibody (Mab 2C6) with strong sperm immobilizing and agglutinating activities was generated by cell fusion between spleen cells from a mouse immunized with human seminal plasma (HSP) and mouse myeloma cells. It also showed a strong inhibitory effect on human sperm-egg interaction. The corresponding antigen was present on the whole surface of ejaculated spermatozoa. In male genital organs, immunostaining with Mab 2C6 was observed in epididymis and seminal vesicle but not in testis. By Western blotting, immunostaining with Mab 2C6 was detected around the 15-25 kDa region under both reducing and non-reducing conditions. The antigen corresponding to Mab 2C6 was susceptible to treatment with periodate or trifluoromethanesulfonic acid. The antigenic activities were slightly increased by treatment with neuraminidase but reduced by further treatment with glycosidases. Enzymatic digestions with pronase and papain also reduced the antigenic activities. The antigen molecules exhibited a strong binding affinity to RCA lectin. These results indicated that Mab 2C6 recognized one of the components which might be secreted from epididymis or seminal vesicle and bind to ejaculated spermatozoa as a sperm coating antigen. The corresponding antigen seems to be a glycoprotein and its carbohydrate moiety has an important role in the conformation of the antigen epitope.

Animals↗

Recognition of carbohydrate antigen epitopes by sperm-immobilizing antibodies in sera of infertile women.

STUDY OBJECTIVE: To study carbohydrate natures in the antigen epitopes corresponding to sperm-immobilizing antibodies in infertile women. DESIGN: Antibody absorption with human sperm and seminal plasma before and after treatments with trifluoromethanesulfonic acid or sodium metaperiodate. PATIENTS: Thirty-three patients who showed a positive sperm immobilization test provided their sera for the experiment. RESULTS: In 25 patients' sera whose sperm-immobilizing antibodies were absorbed with human seminal plasma, the antibody absorbing capabilities were completely abolished by deglycosylation treatment with trifluoromethanesulfonic acid. The sperm-immobilizing antibodies in 4 patients' sera were absorbed out with sperm membrane fraction before the treatment but not after the treatment with trifluoromethanesulfonic acid. In some patients' sera, the antibody-absorbing capabilities of ejaculated sperm were markedly reduced by sodium metaperiodate treatment. CONCLUSION: The majority of sperm-immobilizing antibodies in infertile patients might be generated to carbohydrate structures of the sperm-coating antigens or sperm membrane antigens.

Antibodies↗

Triglyceride and glucose intolerance as a risk factor for coronary heart disease.

The electrophoresis of plasma lipoproteins frequently showed midbands between beta- and pre-beta-lipoproteins in survivors of myocardial infarction. There were increases in intermediate-density-lipoprotein (IDL) cholesterol and triglycerides with an increase in IDL cholesterol/triglycerides in the very-low-density-lipoprotein fraction, even if the increase in cholesterol was not so significant. Impaired glucose tolerance (IGT) was also frequently found in these patients. Among the patients with an apparently normal glucose tolerance, the coronary atherosclerosis scores judged by the American Heart Association reporting system on coronary angiography increased as the total insulin area by 75 g oral glucose tolerance test increased. The correlation between the atherosclerosis score and the insulin area was still significant even after adjustment for body mass index and plasma triglyceride level. Our data suggest that hypertriglyceridemia, IGT, and hyperinsulinemia may become independent risk factors for atherosclerosis in addition to cholesterol. Another study showed that the morbidity of coronary heart disease increased with an elevation of plasma triglyceride levels among patients with familial hypercholesterolemia.

Aged↗

[A clinical study on renal cell carcinoma].

A retrospective study was carried out on 95 patients affected with renal cell carcinoma (RCC) between January 1973 and December 1988. Male to female ratio was 3 to 1 and the average age was 61 years. Hematuria was the most common initial symptom (44 patients, 46.3%), followed by incidental findings (18 patients, 18.9%). Five- and 10-year survival rates for all patients with RCC were 51.9 and 37.1%, respectively. Five-year survival rates for stage I, II, III, and IV were 82.7 54.7, 20.0, and 4.7%, respectively. Statistical analyses identified variables that significantly influenced survival, including lymph node metastasis (p less than 0.01) and distant metastasis (p less than 0.01). Cell type and renal vein involvement did not influence survival significantly. The three different forms of therapy (mitomycin therapy vincristine therapy and immunological therapy) did not influence survival significantly, but the beneficial effect of VBL therapy was obtained.

Carcinoma, Renal Cell↗

Auditory evoked potentials in the Japanese monkey.

Auditory sensitivity based on auditory brain stem response (ABR), whole nerve action potential (AP), and cochlear microphonics (CM) to tone bursts of 0.5-8 kHz were compared with behavioral audiometry in the Japanese monkeys. Although sensitivity loss at 4-6 kHz was observed in these potentials, an increase in sensitivity at 8 kHz was obtained only in the ABR. Thus the sensitivity loss at 4-6 kHz originates at the peripheral system and the increased sensitivity at 8 kHz originates at the central.

Action Potentials↗

[A clinical investigation on renal pelvic and ureteral tumors].

The 60 cases of primary renal pelvic and ureteral tumors treated at Mie University hospitals between January 1977 and December 1987 were reviewed and factors predicting the prognosis were investigated. The patients consisted of 47 men and 13 women (3.6: 1.0). Their ages ranged from 38 to 82 years with a mean of 65.2 years. According to Akaza's category classification of the ureteropelvic tumor, 42 cases were classified to category A, 15 cases category B and 1 case was classified to category C. Histologically, 59 transitional cell carcinomas and 1 squamous cell carcinoma were found. As to grading, 5 was G1, 31 G2, 21 G3 and 2 GX. As to staging, 20 were pT1, 10 pT2, 21 pT3, 3 pT4 and 6 pTX. Staging was correlated well with grading. Total nephroureterectomy with bladder cuff was performed on 39 patients and the other surgical treatments were done on 15 patients. Recurrence of the bladder tumor was found in 22.4%. The 5-year survival rate (Kaplan-Meier's method) was 47.8% for all of the patients. Among the patients with transitional cell carcinoma, the 5-year survival rate was 100% for G1, 57.6% for G2 and 28.6% for G3. As to staging the 5-year survival rate was 90.0% for below pT1, 20.0% for pT2 and 41.1% for pT3. The results from the present study suggest the prognosis is decided by grade and stage in pelvic and ureteral tumors, and it is wanted to develop a system of postoperative adjuvant therapy.

Adult↗

[A retrospective survey of newly registered pulmonary tuberculosis patients--379 cases at 4 health centers in 1985].

Using the registration card of tuberculosis patient, a retrospective analysis was made on newly registered 379 patients with pulmonary tuberculosis in 1985 to examine the treatment duration, the outcome, the duration of registration, and bacteriological relapse. These patients were divided into three groups according to the results of bacteriological examination at the time of registration: (1) patients with positive-bacilli (Group A); (2) patients with negative-bacilli (Group B); and (3) patients without results of bacteriological examination (Group C). 1) Number of cases were 164 (43.3%) in the group A, 130 (34.3%) in the group B, and 85 (22.4%) in the group C. Sputum specimens from 68 patients (80.0%) of the group C were negative on smear, however, the results of culture were not obtained, and remaining 17 cases (20.0%) were not examined bacteriologically. 2) 54 patients (32.9%) in the group A, 25 (19.2%) in the group B, and 17 (20.0%) had a previous history of tuberculosis, respectively. 3) Number of patients with cavitary lesions at the time of registration were 105 (64.0%) in the group A, 46 (35.4%) in the group B, and 14 (16.5%) in the group C. Patients in the group A showed the highest rate of cavitary lesions. 4) Number of patients receiving treatment less than 12 months were 73 (53.3%) in the group A, 92 (85.2%) in the group B, and 47 (73.5%) in the group C, respectively. 5) 317 patients (83.6%) were omitted during 5 years of period from registration although 62 (16.4%) were still under registration even after 5 years.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[A case of type I hyperprolinemia associated with photogenic epilepsy].

A 9-year-old girl with type I hyperprolinemia, who also had photogenic epilepsy, was reported. She showed epileptic discharges and the regression in speech and motor activities, since 7 years of age. Her plasma proline levels were 3 to 4 times higher than control levels. In urine, iminoglycinuria appeared, when plasma proline value exceeded 0.80 mM. The proline oxidase activity of the liver tissues obtained by biopsy in the patient was about 23.5%, compared to that of controls. In spite of the restriction of proline and protein intake, she showed progressive speech and motor retardation.

Amino Acid Metabolism, Inborn Errors↗

Application of the avidin-biotin-peroxidase complex technique for ultraimmunocytochemical characterization of leukemic cells.

For simultaneous demonstration of cellular ultrastructure, myeloperoxidase activity, and presence of a membrane-bound antigen in a given blood cell, we examined three different fixatives: periodate-lysine-paraformaldehyde (PLP) and paraformaldehyde and glutaraldehyde for their applicability to preembedding electron microscopic immunocytochemistry using monoclonal antibodies and the avidin-biotin-peroxidase complex (ABC) technique. This procedure was examined in samples from 3 normal volunteers and 29 patients with acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), lymphosarcoma cell leukemia (LSCL), blastic phase of chronic myelogenous leukemia (CML-BC), or other unclassified leukemias. PLP fixation preserved the immunoreactivity of surface glycoproteins as well as immunoglobulins to the most satisfactory extent. Leukemic cells fixed with PLP maintained their fine structural details, so that we could identify their cytoplasmic organelles, although glutaraldehyde produced the best preservation of cellular ultrastructure. In three patients with ALL, our method revealed that a significant portion of blasts possessed both lymphoid surface antigens and peroxidase-positive cytoplasmic granules. Our method was also useful in identifying the lineage of peroxidase-negative leukemic cells, including monoblastic leukemia and megakaryoblastic leukemia cells. Ultraimmunocytochemistry using PLP fixation and the ABC technique may be a promising strategy for determining the nature of blastic cells that remain unclear after a conventional work-up, for characterizing leukemic cells in patients with a relatively low blast cell count in the bone marrow or peripheral blood, and for estimating the presence and frequency of leukemia with multilineage expression.

Antigens, Neoplasm↗

Detection of interleukin-6 (IL-6) in human bone marrow myeloma cells by light and electron microscopy.

We used an Avidin-Biotin peroxidase complex (ABC) immunoperoxidase technique to evaluate the localization of IL-6 of human bone marrow cells in multiple myeloma (MM). The cellular distribution of IL-6 was determined at light and electron microscopic levels. The author's study indicated that cytoplasmic IL-6 was detected only in the myeloma cells of bone marrow cells. Immunoelectron microscopic (immuno-EM) study showed positive reactivity mainly in the perinuclear space (PNS), well-developed rough endoplasmic reticulum (RER), and Golgi area.

Bone Marrow↗

In vitro endocytosis of benign and malignant human bone marrow plasma cells.

We report ultrastructural evidence of the phagocytic potential of plasma cells and myeloma cells. The incubation of plasma cells and myeloma cells in vitro with horseradish peroxidase (HRP) and cationized ferritin (CF) allows the tracing of fluid-phase and receptor-mediated pathways. Surface-bound ligands (CF) and solutes (HRP) taken up in primary pinocytic vesicles are internalized to the endosomal compartment. After 1 hr of incubation, CF was found not only in plasma cells but also in myeloma cells. Reaction products of HRP were observed only in myeloma cells. In myeloma cells, however, HRP was located only in the lysosomal system, whereas CF was present within membrane cisternae as well as within lysosomes. These myeloma cells morphologically produced interleukin-6 (IL-6).

Bone Marrow↗

Monoclonal antibody recognizing an apparent peptide epitope of human seminal plasma glycoprotein and exhibiting sperm immobilizing activity.

A hybridoma (3B2-F7) has been established which secretes a monoclonal antibody (Mab) directed against a peptide determinant of human seminal plasma glycoprotein (HSP-gP). The deglycosylation of HSP-gP was performed chemically with TFMS hydrolysis and enzymatically in the presence of detergent and further treated with periodic acid after fixing deglycosylated HSP on plastic wells. The Mab 3B2-F7 (IgM, kappa) exhibited sperm immobilization activity (256 units of SI50) and inhibited sperm binding to human zona pellucida. Human epididymis, pancreatic islets of Langerhan's and distal tubulus of kidney were strongly labelled whilst other tissues were essentially negative by avidin-biotin complex tissue staining with this Mab. The antigen epitope to the Mab was in the 36 kDa molecule of human HSP-gP. The antigenic determinant recognized by Mab 3B2-F7 was destroyed by six different proteases, but was resistant to N-glycanase and other carbohydrate splitting enzymes. This epitope is therefore likely to be composed of a polypeptide chain. Peptide fragments after proteolysis of the HSP molecule with Staph. aureus V8 protease and trypsin retained antigenicity, hence the epitope corresponding to the Mab may be a peptide chain and not dependent on the conformational structure of the polypeptide.

Animals↗

Studies on insulin-like substances in Korean red ginseng.

Korean red ginseng powder was found to contain adenosine and an acidic substance which inhibited epinephrine-induced lipolysis and stimulated insulin-mediated lipogenesis from glucose (3, 4). In the present experiment, the chemical structure of this acidic substance is determined to be pyro-glutamic acid. Pyroglutamic acid exhibits selective modulations toward the opposite metabolic pathways in rat adipocytes; it inhibits the lipolysis but rather stimulates the lipogenesis. Based on these results, we suggest to call these substances (adenosine and pyro-glutamic acid) "selective modulators".

Adenosine↗

Proposed distal margin for resection of rectal cancer.

To determine the adequate distal margin, particularly from the point of extent of lymph node metastasis, 2,333 lymph nodes from 44 patients with rectal carcinoma were evaluated, using a clearing method. The tumors were divided into two growth patterns; infiltrative and localized. Lymph node metastasis was histo-pathologically examined with special attention focused on nodes on the distal side of the tumor. The intramural spread was also given attention. The proximal pararectal lymph nodes often contained malignant cells, whereas the distal ones were rarely involved, and if involved, they were present within 1 cm from the tumor. Pararectal lymph node metastasis and intramural spread were seen with a range of 1 cm and 0.5 cm, respectively, in the localized type and 1 cm and 2.1 cm, respectively in the infiltrative type. Based on these findings, the distal margin for surgical resection of rectal carcinoma is considered to be 2 cm for the localized type and 3 cm for the infiltrative type.

Adult↗

[Follow-up study of short course chemotherapy of pulmonary tuberculosis complicated with diabetes mellitus].

A retrospective analysis was made for 644 patients with pulmonary tuberculosis newly diagnosed during the time of 1977 to 1985 to evaluate the influence of diabetes mellitus (DM) on the drug response rate and the long-term relapse rate in the treatment of tuberculosis. These patients were divided into four groups: (1) 123 patients with DM on 9- to 12-month short course regimens; (2) 79 with DM on 13- to 36-month long-term regimens; (3) 379 nondiabetic patients on short-term regimens; and (4) 63 nondiabetics on long-term regimens. Bacteriological relapse after chemotherapy was defined as positive cultures growing at least 20 or more colonies. 1) Bacteriological negative conversion rates were similar in both diabetic and nondiabetic patients who had received combination regimens including INH and RFP. The degree of control of DM did not affect the conversion rate. 2) Of 297 patients who had received short-course chemotherapy and the information for analysis were available as of August 1989, 8 (10.3%) had relapses in 78 diabetic, and patients 23 (10.5%) in 219 non-diabetic patients; the difference was not statistically significant. There were also no discernible differences in the relapse rates between patients on the short-course regimens and those on the long-term regimens. 3) Most of the relapses occurred around 6 months and 30 months after completing the short-course chemotherapy. Similar pattern of relapses was observed also in the long-term therapy group. 4) Pretreatment radiographic findings and quantity of the acid-fast bacilli in the sputum, and the presence of cavitary lesions at the completion of therapy bore no significant relation to the development of relapse. 5) In the diabetic patients the degree of control of DM contributed little to the development of recurrence. 6) Although most of the patients without DM relapsed with sensitive strains and achieved a good response to retreatment, diabetic patients frequently relapsed with resistant strains and had a grave prognosis. 7) Of 41 patients who died after having been on the short-course regimens, two were attributed to tuberculosis. Only one was attributed to tuberculosis of 25 patients who died after receiving the long-term regimens. 8) The data obtained here confirmed that the 12-month regimen for diabetic patients could achieve favorable results in the response rate and the long-term relapse rate, as that for nondiabetic patients. However the cases of tuberculosis complicated with DM frequently showed a poor prognosis once the relapse took place. These results suggest that pyrazinamide-containing 4-drug combination regimens in an initial intensive phase is the preferred treatment for the patients with DM.

Adult↗

[Two cases of X-linked ichthyosis associated with myopathies].

X-linked ichthyosis is an inborn error of metabolism due to the deficiency of steroid sulfatase. We reported two cases of the patients associated with myopathies, which are Duchenne muscular dystrophy (DMD) and myotonic dystrophy (MyD), respectively. In addition to DMD and MyD, they showed corneal opacities, lack of steroid-sulfatase activities in peripheral leukocytes and massive accumulation of cholesterol sulfate in plasma. Such cases were not reported, previously. Assay of steroid sulfatase and cholesterol sulfate in the patients having ichthyosis is important to elucidate the wide clinical spectrum of steroid sulfatase deficiency.

Adult↗