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Biomedical subjects

R Kobayashi

Publications and source records attributed to R Kobayashi.

At least 379 records · Page 21Linked to original sources
PubMed1989

[Biochemical and immunological properties of 34kDa EDTA-extractable protein (EEP34k)].

It has been reported that the 34kDa protein of EDTA-extractable protein (EEP34k) cross-reacted with lipocortin antisera. The lipocortins are a family of proteins that have been reported to inhibit phospholipase A2. Previously, we purified two proteins from EEP34k by anion-exchange chromatography, and referred to them as 34k (A) and 34k (B). On Western-blot analysis, two proteins showed cross-reactivity to both lipocortin I and lipocortin II. In this study, four proteins (34k (A)1, 34k (A)2, 34k (B)1, 34k (B)2) were isolated from 34k (A) and 34k (B), on a cation-exchanged chromatography. By peptide mapping, immunological analysis and SDS/PAGE, we confirmed that 34k (A)1 and 34k (B)2 were identified proteins and these were called lipocortin I, and that 34k (A)2 was lipocortin II. It is concluded that at least two lipocortins, I and II are present in EEP34k. We also observed the presence of phospholipase A2 inhibitory activity in both 34k (A) and 34k (B).

Animals↗
PubMed1989

[A carcinoma of the right breast arising after a mastectomy in primary malignant lymphoma of the left breast].

In 1987, a 51-year-old woman visited our hospital, having noticed a painless 1.5 x 1.5 cm tumor of the left breast. On biopsy, it was pathologically diagnosed as being a primary malignant lymphoma of the breast. Thus we performed a modified radical mastectomy (Patey's method), in combination with chemotherapy (Cyclophosphamide, Vincristine, Prednisolone). Two years after the operation, a tumor of the right breast, 1.0 x 1.0 cm in size, was noticed. At this time, since an excised specimen was pathologically determined as being a scirrhous carcinoma, we performed a modified radical mastectomy (Auchincloss' method), followed by chemotherapy (Tegafur).

Adenocarcinoma, Scirrhous↗
PubMed1989

[Cutaneous malignant lymphoma in childhood].

Two cases of malignant lymphoma in childhood were studied. The first case was a Japanese girl aged 8, in whom the primary site was skin of the right temple. The second case was a 4-year-old Japanese boy, who had metastases to the abdominal skin. Histochemical findings indicated B-cell lineage in both cases. Primary cutaneous lymphoma is extremely rare in childhood. Fourteen such cases that have been reported in Japan and our case added to them, were reviewed. The relationship between their morphologic, immunohistochemical and clinical findings were summarized and discussed. Although the prognosis of lymphoma confined to skin in childhood has been reported not to be bad as compared with other types of lymphoma, our first such case was fatal. This suggests that appropriate initial treatment is very important. Recent advances in science may clarify the clinical and biologic characteristics of this tumor in the near future.

Abdominal Neoplasms↗
PubMed1989

[Application of a nonisotopic southern hybridization method for sex determination from bloodstains found in criminal spots].

Bloodstains collected from criminal spots were submitted to the nonisotopic Southern hybridization (NISH) method for sex identification (Kobayashi et al., J. Forens. Sci., 33, 613-620, 1988). A RNA probe, transcripted from a Y-chromosome specific DNA sequence (Nakahori et al., Nucl. Aci. Res., 14, 7569-7580), was used for this series of investigations. Among the 95 bloodstain specimens studied, in which the sex of each was previously known, 54 (88.5%) of 61 male stains were appropriately determined to be male by the NISH method, and 28 (82.4%) of 34 female bloodstain specimens were estimated to be female. The sex of the 13 remaining stain specimens were unable to be determined by this method, but no specimen was erroneously misidentified. With regard to 12 specimens, in which the sex of each was unknown at the time of the NISH test, 5 were determined as being male and 3 were estimated as being female, though the sex of the remaining 4 specimens could not be determined. The causes in the stains that made determination impossible were analyzed using patterns of ethidium bromide fluorescence of electrophoresed gel and/or NISH, and it was concluded that this could be due to an insufficient quantity of extracted DNA (12 cases), the resistance against a restriction enzyme (EcoRI) (3 cases), and the depolymerization of the Y-chromosome specific fragment (2 cases).

Blood Stains↗
PubMed1989

Sex determination of bloodstains by the use of a ribonucleic acid (RNA) probe.

An asymmetric ribonucleic acid (RNA) probe, which represents a Y-chromosome specific nucleotide sequence, was applied to the sex origin of bloodstains by using a nonisotopic Southern hybridization method. The RNA probe was in vitro transcribed from the recombinant of the Y-chromosome specific deoxyribonucleic acid (DNA) sequence and transcription-vector Bluescript SK M13+. The use of this RNA probe can lower the detection limit of the nonisotopic DNA analyzing method to 1/8 of that reached by our previously reported method in which a DNA probe was used.

Blood Stains↗
PubMed1988

Isolation of endopeptidase-24.11 (EC 3.4.24.11, "enkephalinase") from the pig stomach. Hydrolysis of substance P, gastrin-releasing peptide 10, [Leu5] enkephalin, and [Met5] enkephalin.

The purpose of this investigation was to isolate the cell-surface enzyme endopeptidase-24.11 from the stomach wall of the pig and to examine the hydrolysis of the gastric neuropeptides. Endopeptidase-24.11 was isolated from gastric membranes by immunoadsorbent chromatography using a monoclonal antibody to porcine kidney endopeptidase-24.11. The enzyme was purified with a yield of 1.2 micrograms/g wet wt of fundic muscle. A single polypeptide chain of apparent subunit molecular weight of 90,000 was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gastric endopeptidase-24.11 hydrolyzed substance P, gastrin-releasing peptide 10, [Leu5] enkephalin, and [Met5] enkephalin by cleavage of peptide bonds on the N-terminal side of hydrophobic amino acids. The enzymatic activity was inhibited completely by phosphoramidon (10(-6) M) and strongly by 1,10-phenanthroline (10(-3) M), but was unaffected by captopril (10(-5) M).

Animals↗
PubMed1988

The perimenarche syndrome (a proposal).

We present here two cases with depression and obsessional symptoms, in which the patients were 11-year-old girls just prior to the advent of menarche. Their clinical picture was fairly different from the one which Deutsch, H., Jacobson, E. and Blos, P. described in their classical cases so far. In our cases the regression to preoedipal level was very prominent, that is, the rapprochement crisis-like state (Mahler, M.S.) with transitional objects such as stuffed animals and dolls (Winnicott, D.W.) was observed. It was believed that with such a background there were changes in the parent-child relationship in preadolescence which the premature physical growth among recent children had brought about, and changes in the concept of the mother and father which cultural vicissitudes had induced. It was also indicated that this kind of direct observation would contribute not only to the encouragement of the sound development in this period of life, but also the understanding and treatment of adolescent or adult cases.

Anorexia Nervosa↗
PubMed1988

Metabolism of gastrin and cholecystokinin by endopeptidase 24.11 from the pig stomach.

The purpose of this investigation is to examine the metabolism and inactivation of human and porcine gastrin 17 (nonsulfated) (G-17) and cholecystokinin octapeptide (sulfated) (CCK-8) by gastric endopeptidase 24.11. Endopeptidase 24.11 was isolated by immunoaffinity chromatography using a monoclonal antibody to the kidney enzyme. Peptides were incubated with endopeptidase 24.11. The digests were either fractionated by reverse-phase high-pressure liquid chromatography and the products identified by amino acid analysis or they were used for bioassays. Digests of human gastrin were assayed for stimulation of acid secretion in the anesthetized rat, and cholecystokinin digests were assayed for the stimulation of amylase secretion from isolated rat pancreatic acini. Human G-17 was degraded by cleavage of the Trp4-Leu5,Ala11-Tyr12,Gly13-Trp14,Trp14 -Met15, and Asp16-Phe17-NH2 bonds, and the fragments (1-16), (1-13), (1-11), (1-4), (5-11), (5-13), (12-13), (12-14), (14-16), and (17-NH2) were identified. Porcine G-17 was degraded by hydrolysis of the Ala11-Tyr12,Gly13-Trp14, and Asp16-Phe17-NH2 bonds producing (1-16), (1-13), (1-11), (12-13), (14-16), and (17-NH2) fragments. CCK-8 was degraded by hydrolysis of the Gly4-Trp5 and Asp7-Phe8-NH2 bonds, and the fragments (1-7), (1-4), (5-7), (5-8), and (8-NH2) were identified. There was a progressive decline in the biological activity with incubation time.

Amino Acid Sequence↗
PubMed1988

Sex identification in fresh blood and dried bloodstains by a nonisotopic deoxyribonucleic acid (DNA) analyzing technique.

Deoxyribonucleic acid (DNA) specimens were prepared from blood or bloodstain extracts, and the content of a Y-chromosome specific DNA fragment was investigated by the Southern hybridization method using a nonisotopic staining technique. Thus obtained patterns of male DNA showed a clear band, whereas broad stains with some faint bands appeared on the patterns of DNA from both sexes. This method is expected to be a new powerful mean of forensic medical examination.

Autoradiography↗
PubMed1988

Autoantibody against erythrocyte protein 4.1 in a patient with autoimmune hemolytic anemia.

We observed the presence of a new autoantibody, anti-erythrocyte protein 4.1, in a patient with autoimmune hemolytic anemia (AIHA). Western blotting analysis revealed that IgG from the patient's plasma reacted with erythrocyte protein 4.1. However, among other patients with hemolytic diseases (six having AIHA and three each having either hereditary spherocytosis, elliptocytosis, or lead poisoning) as well as among control subjects, no antibody activity to protein 4.1 was observed. In addition to the anti-protein 4.1 antibody, two different kinds of anti-erythrocyte antibodies were detected by conventional serological studies in this patient. One of them was an anti-Ena-like antibody in the eluate from the patient's erythrocytes, while another was the anti-S-specific antibody in the plasma. An elution study and an absorption study using S antigen-positive erythrocytes demonstrated that the anti-protein 4.1 antibody differed from both the anti-Ena-like antibody and the anti-S antibody. Familial analysis of the patient revealed the same antibody in her brother, who did not have hemolytic anemia. These results demonstrate that anti-protein 4.1 antibody is considered to be included in the spectrum of anti-cytoskeleton autoantibodies, which have been observed in patients having increased cell lysis as well as in healthy subjects.

Anemia, Hemolytic, Autoimmune↗
PubMed1987

Neurotensin-like immunoreactivity generated by pepsin from human plasma and gastric tissue.

The present study demonstrates precursors of neurotensin-like immunoreactivity (NTLI) endogenous to human gastric tissue and plasma, and the existence of a gastric NTLI-generating enzyme system. The molecular size of the NTLI-precursors in plasma and gastric tissue were estimated by gel permeation chromatography to be ca 50,000-60,000 and 60,000-70,000 Da, respectively. The neurotensin-like peptide generated from the precursor was detected with a carboxyl-terminally directed antiserum but did not cross-react with an amino-terminally directed antiserum. A neurotensin-like peptide isolated from pepsin-treated human plasma was characterized by mass spectrometry and its amino acid sequence determined. This novel nonapeptide, referred to as kinetensin, failed to affect pentagastrin-stimulated acid secretion or blood pressure in the rat. Sequence homologies between neurotensin, kinetensin and proteins of the serum albumin family suggest a common evolutionary origin and raise questions regarding albumin-like proteins as precursors of regulatory peptides.

Amino Acid Sequence↗
PubMed1987

Immunohistochemical studies on GABAergic neurons in the rat locus coeruleus, with special reference to their relationship to astrocytes.

Purified antisera against GABA were prepared. A few small GABAergic neurons in the rat locus coeruleus were immunohistochemically demonstrated by both the unlabeled peroxidase-antiperoxidase method and the avidin-biotin peroxidase complex method using affinity-purified GABA antibodies. The glial fibrillary acidic protein immunoreactivity in this nucleus was localized by the latter method in the astrocytal framework encircling medium-sized and small neurons as well as in straight processes. Astrocytes may play a role as energy donors to these neurons.

Animals↗