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Biomedical subjects

M Kikuchi

Publications and source records attributed to M Kikuchi.

At least 847 records · Page 47Linked to original sources
PubMed1989

Luteal hypoplasia and in vitro granulosa cells proliferation in cows.

Two groups of cows were set up. In one group ovulation was induced, leading to luteal hypoplasia (FSH non-injection group or group 1). In the other group ovulation was induced and FSH injected simultaneously to improve luteinization (FSH injection group or group 2). Granulosa cells were collected from the ovarian follicles of the cows of the two groups immediately before ovulation. Their ability to proliferate and secrete progesterone in vitro was compared between the two groups. Granulosa cells proliferated in vitro more rapidly in group 2 than in group 1. Therefore, FSH injection was proved to be very effective for the proliferation of these cells. Cultured granulosa cells showed the same morphology, regardless of the FSH injection. They presented changes from fibroblast-like cells to epithelioid cells after adhering to a coverslip. They were transformed to epithelioid cells, however, earlier in group 2 than in group 1. Their ability to secrete progesterone in vitro was essentially the same between the two groups. Progesterone secretion decreased gradually in amount in both groups with the advance in culture. Therefore, it was considered that luteal hypoplasia might have been caused by a low ability of granulosa cells, which would be transformed to lutein cell, to proliferate.

Animals↗
PubMed1989

Morphometrical and biochemical differences of endocrine pancreata between spontaneously hypertensive and normotensive rats with or without neonatal streptozotocin-induced diabetes.

We studied the morphometrical and biochemical changes of endocrine pancreata in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) with or without noninsulin-dependent diabetes mellitus induced by neonatal streptozotocin (STZ) treatment at 4 months of age. Female (2-day-old) neonates were intraperitoneally injected with 62.5 or 75.0 mg/kg of STZ for SHR, 87.5 or 100.0 mg/kg of STZ for WKY, and vehicle for control. In STZ-treated groups, overt hyperglycemia developed in SHR with significantly decreased serum immunoreactive insulin (IRI), whereas in WKY, hyperglycemia was very mild and serum IRI was not lowered. The number and mean size of pancreatic islets did not differ between SHR and WKY, although mean islet size was reduced by half in both compared with that in the corresponding control, respectively. Percentage distribution of insulin-positive B cells in the islet was significantly reduced more in SHR than in WKY (34% of control versus 64% of control, p less than 0.05). Furthermore, pancreatic IRI content was far more reduced in SHR than in WKY (3% of control versus 43% of control, p less than 0.001). In vehicle-treated groups, the glycemic levels and the morphometrical islets did not differ between SHR and WKY. However, serum IRI was significantly lower but pancreatic IRI content was higher in SHR than in WKY. The mechanisms of strain differences between SHR and WKY seen in the present study were discussed.

Animals↗
PubMed1989

Analysis of T-cell receptor delta chain gene in hematological malignancies.

We analyzed the rearrangement of TcR delta chain gene in 179 cases of hematological malignancies. In 17 T-cell lines, RPMI 8402, DND41, Peer, and Molt 13 had delta rearranged band (s). Except for RPMI 8402, these cell lines expressed functional delta gene. All of those gamma delta-T-cell lines had short message (1 kb) of TcR beta gene. These findings suggest differences between alpha beta-T-cells and gamma delta-T-cells. All 9 cases of T-ALL/LBL, of which 4 had neither gamma nor beta gene rearrangement, had a new rearranged band of TcR delta locus. This rearrangement was observed in 63% of B-lineage ALL/LBL. In the other T-lymphoproliferative disorders, only 2 cases of AILD and 1 of T-cell lymphoma had the rearranged band (s), showing derived T-cell neoplasm from gamma delta-T-cell as minority. In B-leukemia/lymphoma and myelocytic leukemia, 15% of the cases had the delta rearrangement. Heterogenous findings of TcR delta locus analysis were observed in ATLL without proviral HTLV-I DNA, T-cell lymphoma, AILD and HD. The J delta 1 region was frequently used and the J delta 2 region was rearranged in one AILD. It is suspected that J delta 3 was used in one T-ALL/LBL. There was no correlation between the phenotypic pattern of CD3, CD4, and CD8 in T cell disorders and the rearrangement of the TcR delta gene. These findings suggest that the newly identified TcR delta chain gene rearranges at a very early stage of T cell ontogeny; prior to the other TcR genes and perhaps at almost the same differentiation level as that of CD7 expression. The TcR delta gene is useful in evaluating clonality for the most immature T cell neoplasms not showing rearrangement of the other TcR genes. This gene is not lineage specific, however, when used in conjunction with IgHC gene, it may be a useful tool for the study of ALL/LBL.

Gene Rearrangement, T-Lymphocyte↗
PubMed1989

[Secondary leukemia (AML) with TcR beta rearrangement].

A 60-year-old female with fever and general lymphadenopathy was admitted to our hospital in 1979. By histological examination of cervical lymph node, she was diagnosed as B-cell malignant lymphoma, diffuse, large, stage IV B (Ann Arbor classification). In 1984, the complete remission of malignant lymphoma was observed after MOPP, COPP and VAPP chemotherapies. After 3 years, she was readmitted because of leukocytosis in which 64% immature abnormal cells were detected. By conventional cytological and cytochemical evaluations, the immature abnormal cells were identified as secondary AML cells with M 2 type (FAB), peroxidase (+), My 7 (+) and chromosomal abnormality; 48, X, -X, +4, -9, +21, del (1) (p 34), i (17 q), + M 1, + M 2. Analysis of Ig and TcR genes showed TcR-beta rearrangement. It has been reported that secondary leukemia frequently causes chromosomal abnormalities and therapy resistance. Ig gene and TcR gene analysis of secondary leukemia combined with chromosome examination, which has reported little, may be useful in predicting prognosis and therapy resistance.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1989

[A study of rheumatoid arthritis patients associated with biopsy-proven secondary amyloidosis].

Reactive systemic amyloidosis associated with rheumatoid arthritis (RA) was studied clinically in 28 patients (2 men and 26 women). The diagnosis of amyloidosis was established by histological examination of biopsy materials. Upper gastrointestinal tract biopsy was performed in 14 patients, and renal and rectal biopsy in 8 and 4 respectively. The mean age and duration of RA at diagnosis of amyloidosis were 58.6 (range 35-72) years and 15.5 (range 4-44) years respectively. Almost all patients had intractable and progressive courses of RA. Serological activities determined by C-reactive protein (CRP) and erythrocyte sedimentation rates were moderate to high in over 80% of the cases. Renal abnormalities were noticed in 19 cases, and gastrointestinal disorders in 10. Eight patients died from 1 to 54 (mean 15.3) months after the diagnosis of amyloidosis; 5 died of renal failure and 2 of gastrointestinal involvements. Renal impairments progressed frequently and serum creatinine elevated over 1.5 mg/dl in another 8 cases. Five patients progressing to renal failure were treated with hemodialysis. Three died within several weeks after the induction of hemodialysis, although 2 were treated for more than 2 years. Intractable hypotension and pulmonary congestion were frequently observed in these cases. A close relationship was found between serum amyloid A protein (SAA) and CRP concentration, so that the measurement of SAA seemed to be valuable in assessing disease activity. Concerning the treatment of amyloidosis, cyclophosphamide and corticosteroids seemed to be effective in several cases, although it had been unsatisfactory in most cases.

Adult↗
PubMed1989

[Successful treatment with a small dose of N4-behenoyl-1-beta-D-arabinofuranosylcytosine in a case of chronic myelomonocytic leukemia in blastic crisis].

A 56-year-old female was admitted for the examination of leukocytosis in May, 1982. The hematological examination showed hemoglobin 12.0g/dl, platelets 14.5 X 10(4)/microliters and leukocytes 18,000/microliters with 28% of monocytes. A bone marrow aspiration revealed granulocytic hyperplasia. Granulocytes showed nuclear abnormalities, such as folding or lobulation. From these data, a diagnosis of chronic myelomonocytic leukemia (CMMoL) was made and followed with no treatment for 2 years. However, fever, bone pain, anemia, thrombocytopenia and the increase of monoblasts in the peripheral blood and bone marrow were observed in May, 1984. These findings indicated that she was in the blastic phase. She was treated by intensive combination chemotherapy (BHAC-AMP), but did not attain any remission. Therefore, a small dose of N4-behenoyl-1-beta-D-arabinofuranosylcytosine (BH-AC; 1 mg/kg/day) was administered for 70 days. As a result, complete remission was obtained and continued for 37 months with the same therapy. The experience of this case suggests that small dose of BH-AC could be applied to a case of CMMoL in blastic crisis.

Antineoplastic Agents↗
PubMed1989

Very well differentiated adenocarcinoma of the colon: a case report.

A 47-year-old woman with chronic colitis was found to have a sessile tumor at the hepatic flexure of the colon. An endoscopic biopsy revealed no aytpia, but surgery was performed since, in view of the tumor's size, malignancy could not be ruled out. A tumor measuring 3 x 2 x 1.5 cm was found in the resected specimen. The tumor exhibited quite unusual histologic features, showing invasive, moderately atypical glands forming mucin-filled cysts, very similar to those seen in colitis cystica profunda, under pseudopolypoid mucosa covered by non-atypical glandular epithelial cells. The extension of glandular elements into the pericolonic fatty tissue, and the intermingling of anaplastic microglands led to a final diagnosis of adenocarcinoma. In a case such as this, it is very difficult to evaluate cancerous tissue by endoscopic biopsy, even with an amount of tissue adequate for histologic examination.

Adenocarcinoma↗
PubMed1988

Soluble dipeptidyl peptidase IV from terminal differentiated rat epidermal cells: purification and its activity on synthetic and natural peptides.

In terminally differentiated epidermal cells dipeptidyl peptidase IV (EC 3.4.14.5) (DPP IV) is present mainly in a soluble form. We purified the enzyme from 2-day-old rat cornified cells to homogeneity by Sephadex G-200 and Mono-Q column chromatography and finally HPLC gel filtration on G3000SW. The enzyme was estimated to be Mr 190,000 by HPLC gel filtration and Mr 90,000 by sodium dodecyl sulfate-electrophoresis. The enzyme showed general properties reported for detergent-solubilized DPP IV from other tissues. It was Con A binding and almost completely inhibited by 1 mM diisopropyl fluorophosphate and Diprotin A. The pI was 5.6 and the pH optimum was 7.5. The specific activity for Gly-Pro-p-nitroanilide was 31.9 units/mg. HPLC analysis demonstrated the release of dipeptides of the N-terminal of substance P, beta-casomorphin, and their related peptides. A stoichiometric reaction of the enzyme on substance P was observed. The epidermal DPP IV had a Km of 0.3 mM and a kcat of 50.3 s-1 for substance P and the Km value decreased by shortening the peptide from the carboxyl-terminal amino acids. The enzyme hydrolyzed human and bovine beta-casomorphin with Km values of 0.025 and 0.05 mM, respectively. Shortening the bovine beta-casomorphin peptide chain did not affect enzyme affinity.

Animals↗
PubMed1988

[The effect of purified mouse GM-CSF on CFU-C colony growth in serum-free cultures].

We investigated the effect of the ingredients in serum-free cultures, in which serum was completely replaced by albumin, cholesterol and transferrin, on the growth of murine granulocyte/macrophage progenitor cells (colony forming unit in culture: CFU-C) stimulated by serum-free PWM-SCM and the influences of purified granulocyte-macrophage colony stimulating factor (GM-CSF) in serum-free cultures. The results were as follows: 1) The number of CFU-C colonies reached a peak after 4 days of incubation. 2) A linear relationship was observed between the number of CFU-C colonies and the number of inoculated bone marrow cells. 3) Serum-free cultures could support CFU-C colony growth to the same degree as that in serum-containing cultures. Also, no significant differences were found in the types of colonies grown in both cultures. 4) Bovine serum albumin (BSA) and cholesterol were considered to play the most important roles among the ingredients in serum-free cultures. 5) Purified GM-CSF supported CFU-C colony growth in serum-free cultures and more than a half of the colonies formed were GM-colonies. These results showed the usefulness of our serum-free cultures for studying the granulopoiesis in vitro without the influences of various substances in the serum.

Animals↗
PubMed1988

Primary structure of non-histone protein HMG1 revealed by the nucleotide sequence.

The isolation and sequencing of a cDNA clone coding for the entire sequence of pig thymus non-histone protein HMG1 are described. The sequence analysis reveals a complete 2192-nucleotide sequence with a 5'-terminal untranslated region of 11 nucleotides, 642 nucleotides of an open reading frame that encoded 214 amino acids, and a 3'-terminal untranslated region of 1539 nucleotides. The HMG1 protein, deduced from the nucleotide sequence, has a molecular weight of 24,785 and a C-terminal of a continuous run of 30 acidic amino acids, encoded by a simple repeating sequence of (GAN)30. The predicted amino acid sequence is homologous to HMG1, HMG2, and HMG-T sequences from several sources, suggesting that the protein conformation is under evolutionary constraints. Northern blot analysis reveals that another hybridizable RNA species of smaller size is present. Southern blot analyses suggest that pig genome contains several HMG1 gene equivalents.

Amino Acid Sequence↗
PubMed1988

Role of disulfide bonds in folding and secretion of human lysozyme in Saccharomyces cerevisiae.

We examined folding and secretion of human lysozyme using four mutants each lacking two cysteines expressed in a yeast secretion system. Our results have revealed that the formation of the disulfide bond Cys6/Cys128 in human lysozyme is a prerequisite for correct folding in vivo in yeast. Substitution of Ala for Cys77 and Cys95 gave eight-fold greater secretion of a molecule with almost the same specific activity as that of the native enzyme. Substitutions of the other cysteines gave molecules that were secreted at a lower rate and had lower specific activities than the native enzyme. These are the first findings that the individual disulfide bonds of human lysozyme have different functions in folding and secretion in vivo.

Chromatography, High Pressure Liquid↗
PubMed1988

Purification and characterization of bradykinin-hydrolyzing enzyme from 2-day-old rat epidermis.

Bradykinin-hydrolyzing enzyme was purified 200-fold from a soluble fraction of cornified cells from 2-day-old rat epidermis. The enzyme has an Mr of 80,000 as identified by SDS polyacrylamide gel electrophoresis and HPLC gel filtration. The isoelectric point of the enzyme is 5.05. The enzyme hydrolyzed Phe5-Ser6 of bradykinin and seven bradykinin-related peptides, and Tyr5-Ser6 of Tyr5-bradykinin. Production of bradykinin fragments, Arg-Pro-Pro-Gly-Phe and Ser-Pro-Phe-Arg, proceeded in a stoichiometric fashion. Km and Vmax values for bradykinin were 33 microM and 22.2 mumol/min per mg, respectively. The enzyme did not hydrolyze azocasein, denatured hemoglobin or synthetic substrates for other epidermal proteinases. The enzyme activity was enhanced by reducing agents and inhibited by sulfhydryl-blocking agents and divalent cations. Diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride had no effects. The enzyme has a pH optimum of 7.0-7.5 and is stable at 4 degrees C for 1 month, but loses activity completely at 60 degrees C for 10 min. The epidermal endopeptidase differs in several properties from endooligopeptidase A purified from brain which hydrolyzes Phe5-Ser6 of bradykinin.

Amino Acid Sequence↗
PubMed1988

Early development of nephropathy in a new model of spontaneously hypertensive rat with non-insulin-dependent diabetes mellitus.

We designed the present study to clarify whether the development of nephropathy was accelerated by a combination of hypertension and non-insulin-dependent diabetes. Spontaneously hypertensive rats with non-insulin-dependent diabetes induced by neonatal streptozotocin treatment (25.0-75.0 mg/kg) were separated into severely or mildly diabetic groups according to their non-fasting plasma glucose levels at 12 weeks of age and the findings were compared with the data on a control group treated with citrate buffer alone. The natural courses of urinary excretion rate of total protein, the molecular composition by sodium dodecyl sulfate polyacrylamide gel electrophoresis with laser densitometer and N-acetyl-beta-D-glucosaminidase were measured in the three groups from 12 weeks until 36 weeks of age. Total urinary protein in the control group decreased with age (p less than 0.05), while in the mildly diabetic group changes were nil; in the severely diabetic group, however, the excretion rates of total urinary protein and high molecular weight protein consistently and progressively increased with age (p less than 0.05). The low molecular weight protein continuously decreased with age in the mildly diabetic and control groups (p less than 0.05), while in the severely diabetic group there was no decrease after 28 weeks of age. The urinary N-acetyl-beta-D-glucosaminidase markedly increased (p less than 0.05) in the severely diabetic group throughout the period compared with findings in the control group, but drastically decreased (p less than 0.05) in the mildly diabetic group with age.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗