Synthetic studies on enkephalin analogs. III. A highly potent enkephalin analog, H-Tyr-D-Met(O)-Gly-Phe-NHNH-CO-CH2CH3.
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Biomedical subjects
Publications and source records attributed to M Fujino.
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A highly specific enzyme immunoassay for determining hCG was established by using beta-D-galactosidase as label. In order to increase the specificity of the assay, an antiserum against whole hCG was purified on a column of hCG beta carboxyl-terminal peptide (residues 123-145) covalently linked to Sepharose 4B. The antibody (N101S) thus prepared showed a weak cross-reactivity with human LH in an assay using hCG-enzyme conjugate, but the slight cross-reactivity was virtually avoided when an hCG beta carboxyl-terminal peptide was used as a peptide in the enzyme conjugate. N101S antibody was compared with antiserum (B1B) directed against a carboxyl-terminal peptide (123-145). In hCG measurement N101S gave about 30 times higher sensitivity than B1B, although the former antibody was less sensitive to carboxyl-terminal peptides of hCG beta. The enzyme immunoassay using a combination of N101S antibody and a carboxyl-terminal peptide (130-145)-enzyme conjugate was able to detect as little as 0.25 mIU of hCG without the interference of LH. The performance and validity of this assay were comparable to those of conventional radioimmunoassay.
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Carcinoembryonic antigen (CEA) activity was confirmed in the C-cell of the normal thyroid by immunohistochemical techniques. This suggests that CEA production in medullary carcinoma of the thyroid is not initiated by carcinogenesis, but reflects a function of the normal C-cell. It is not yet clear whether CEA production in the C-cell may be influenced by carcinogenesis. The C-cell is the first APUD cell that was confirmed to have CEA activity.
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To obtain further evidence concerning the intracellular action of ouabain, we observed the effect of this compound on contractures evoked by rapid cooling in the presence of caffeine (rapid cooling caffeine contracture; RCCC), in cardiac muscles under conditions of a Ca deprived medium. The experiments were carried out at 20 degrees C using cat papillary muscles and frog ventricle strips, with the exception of cooling (2 degrees C). The muscles were first immersed in normal solution, electrically driven, then immersed in 1 mM EDTA solution for 6 min. The strips were then immersed for 10 min in a caffeine solution prepared by eluting a no-Ca solution containing 50 mM caffeine through Dowex A-1, Ca-chelating resin. Finally, the caffeine solution was exchanged with fresh caffeine solution of 2 degrees C to evoke to RCCC. Ouabain (1 X 10(-7)--1 X 10(-5) M) markedly potentiated RCCC. An ouabain-induced increase of RCCC did not appear in the resting strips unless the tissues were electrically stimulated. 3H-ouabain occupied a considerable cellular space (0.55) at the appearance of the ouabain potentiation of RCCC (14C-inulin space; 0.20). The appearance of the ouabain potentiation of RCCC was independent of changes in 22Na, Ca2+ and ATP contents in the strips. A possible mechanism of the potentiating effect of ouabain on contraction is discussed.
The antianginal action of a sustained-release preparation of isosorbide dinitrate (SUS-ID), administered twice a day, was examined in open trial in 61 patients with angina pectoris. The results obtained were as follows: 1) The frequency of anginal attacks as well as consumption of nitroglycerin tablets began to decrease significantly in the two weeks following the commencement of treatment with SUS-ID; increasing the dose and prolonging the period of administration resulted in further augmentation of its clinical effect. 2) The subjective symptoms improved in 44 of the 61 patients studied (72.1%), and 17 patients (27.9%) showed an improved electrocardiogram. 3) Both the systolic and diastolic blood pressures were significantly lowered, but little change in heart rate was observed. 4) According to the efficacy assessment by the doctors in charge, this drug was judged to be "effective and strongly recommendable" and to be "usable" in 56 of 61 patients (91.8%). Thus SUS-ID was considered to be of clinical usefulness. 5) In terms of side effects, 14 patients (23.0%) complained of headache and three patients (4.9%) of dizziness. These side effects, however, were not serious enough to cause withdrawal of the drug.
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The present study was performed to determine the effect of ouabain on Ca release from fragmented sarcoplasmic reticulum (FSR) isolated from cat cardiac muscles. The results clearly demonstrate that ouabain potentiates the Ca release from FSR by changing the ionic environment.
Serum carcinoembryonic antigen (CEA) and calcitonin were assayed in 8 patients with medullary carcinoma of the thyroid (MCT) and 14 unaffected family members, from 4 pedigrees of Sipple's syndrome and one pedigree with inherited MCT. Serum CEA ranged from 5.4 to 44.5 ng/ml in the patients, but less than 2.5 ng/ml in the unaffected. This is in contrast with retinoblastoma and cancer family syndrome, where not only the patients but also unaffected family members show high serum CEA, and cytoplasmic or selective maternal inheritance of serum CEA level is presumed. In inheritable MCT, serum CEA increases in association with the development of MCT, and serum CEA level per se is not inherited.
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Earliest endoscopic appearance and its subsequent changes were studied in 41 cases of early gastric carcinoma where gastrocamera photographs were available for detailed analysis over the period from three months up to 10 years. The earliest appearance of polypoid carcinoma was a small area of nodularity, and that of carcinoma with mucosal depression was patchy reddening. Both types of carcinoma were thought to occur initially as a lesion without mucosal elevation or depression (IIb). In carcinoma with shallow mucosal depression (IIc and IIc-+III), there were cases believed to have remained intramucosal for an extremely long period of several years, whereas, in polypoid carcinoma (IIa and I), the growth seemed to be more rapid. In the cases with ulcer within the carcinomatous lesion, healing of the ulcer was confirmed in 37%, and "malignant cycles" were observed in 21%. Malignant transformation of gastric ulcer was suggested in one case with linear ulcer.
Seven rabbits were immunized with a synthetic C-terminal glucagon fragment [15--29] conjugated with bovine serum albumin by means of glutaraldehyde. Antisera for glucagon were produced in all the animals after six injections of the conjugate. One of them revealed a higher titer antiserum (G42), which did not cross react with gut glucagon-like immunoreactive material, secretin, insulin, gastric inhibitory polypeptide or vasoactive intestinal peptide. From the results of inhibition of 125 I-glucagon in binding with the antiserum by various glucagon-related fragments the immunogenic determinant of the antiserum was proved to be in the C-terminal residue of the glucagon molecule, although peptide [17--29] or [21--29] reacted weakly with the antiserum. The plasma glucagon levels measured by antiserum G 42 during an arginine test in five normal subjects were superposed on those obtained by other antiserum (G21), specific for pancreatic glucagon. Furthermore, a comparable standard curve for glucagon was obtained using antiserum G42, when a labelled p-hydroxyphenylacetylated glucagon fragment [15--29] was employed as a tracer. The present study clearly demonstrated that the C-terminal glucagon fragment could yield a specific antiserum for pancreatic glucagon, supporting the proposal that the C-terminal fragment of glucagon is responsible for such specific antisera. Furthermore, it is concluded that immunoassay for glucagon could be performed using the labelled glucagon fragment as a tracer.