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Biomedical subjects

C Kimura

Publications and source records attributed to C Kimura.

At least 19 recordsLinked to original sources

Expression of human pituitary adenylate cyclase activating polypeptide (PACAP) cDNA in CHO cells and characterization of the products.

cDNA encoding human PACAP precursor was expressed in non-neuroendocrine Chinese hamster ovary cells, CHO-K1, The cells were transfected with expression vector (pTS705) containing the human PACAP cDNA by electroporation. A cell line which produced more than 80 ng/ml of immunoreactive PACAP (ir-PACAP) into the conditioned medium was established. RP-HPLC analysis of culture medium of this established cell line exhibited the presence of two types of PACAP, i.e. PACAP38 and PACAP27. At the same time, it was also revealed that immunoreactive PACAP-related peptide (ir-PRP) was secreted into the cultured medium. The ir-PACAPs were confirmed to ahve biological activities such as induction of cAMP and neurite outgrowth in rat pheochromocytoma PC12h cells.

Adenylyl Cyclases

Structure of the human pituitary adenylate cyclase activating polypeptide (PACAP) gene.

The human gene encoding pituitary adenylate cyclase activating polypeptide (PACAP) was isolated and its nucleotide sequence was determined. By comparison with a human PACAP cDNA, the exon/intron organization of PACAP gene was determined. The last exon encoded the longer form of PACAP, PACAP38 and 3'-untranslated sequences, suggesting that the shorter form of PACAP, PACAP27 is not generated by alternative splicing mechanisms. The 5'-flanking region of the PACAP gene contains several sequence motifs homologous to CRE, TRE, and GHF-1. On the basis of DNA isolated from mouse A9 microcell hybrid clone containing a single human chromosome, the PACAP gene was assigned to human chromosome 18. Furthermore, we determined the locus of the gene to be 18p11 by the chromosomal in situ hybridization technique.

Amino Acid Sequence

Type I allergy to foods in atopic dermatitis. Comparison between RAST-positive and RAST-negative cases.

Radioallergosorbent tests (RASTs) with five common foods were performed in 183 patients with atopic dermatitis. The results showed that about half of the patients had type I allergy to at least one of the five foodstuffs. The RAST results correlated roughly to the severity of dermatitis. In each group of patients with mild, moderate and severe atopic dermatitis, positive RAST reactions to common foods occurred predominantly in patients who had a personal or a family history of respiratory atopy. Positive RASTs were rare in patients with pure atopic dermatitis who had neither personal nor family history of respiratory atopy. It seems likely that type I food allergy occurs predominantly in those patients with atopic dermatitis who have a predisposition to respiratory atopy.

Adolescent

[Clinical study of panipenem/betamipron for burn infections in the domain of plastic and reconstructive surgery].

The effectiveness and the safety of panipenem/betamipron, new antibiotics of the carbapenems for burn infections, were studied and the following results were obtained: 1. The preparation, 0.5 g/0.5 g, was administered by intravenous drip infusion twice a day to 11 cases of patients with burn infections. In 10 cases for which clinical effects were evaluable, results were rated as "excellent" in 2 cases, "good" in 2 cases and "fair" in 6 cases, with an efficacy rate of 40%. 2. Penetration to the affected tissue was studied in 2 cases. The tissue level of panipenem was 0.20 micrograms/g immediately after the end of drip infusion and 6.86 micrograms/g 60 minutes thereafter. 3. As for the safety, a slight increase in GOT, GPT and Al-P was noted in 1 case; a slight increase in GPT, NAG and beta 2MG was found in 1 case; and a slight increase in GOT, GPT, Al-P and LAP was noted in 1 case, as abnormal variations in laboratory test results.

Adolescent

Artery anatomy and tortuosity in the distal finger.

The arterial architecture in the finger distal to the proximal interphalangeal joint was studied in sixty-seven cadaver fingers with the aid of an operating microscope. The course, frequency, location, and diameter of the dorsal nail fold artery and its anastomosis was recorded. Similar measurements were performed for the palmar anastomosis. The frequency of arterial tortuosity in the digital artery was compiled and a classification of its morphology devised. The characteristic of arterial tortuosity provides a degree of protection to these essential structures. Failure to recognize this facet of arterial anatomy may be one more factor contributing to our inability to successfully revascularize the distal finger in certain patients.

Arteries

The distal venous anatomy of the finger.

An exhaustive anatomic dissection of all of the veins of the finger distal to the proximal interphalangeal joint was done. More than 3200 segments of veins were individually dissected out, measured, and recorded. From this survey new schematic diagrams have been drawn emphasizing the pertinent venous anatomy at the proximal, distal interphalangeal joints and eponychial levels. Suggestions are made for regions that are apt to have the largest vessels available for anastomosis.

Cadaver

Prevention of the incidence of diabetes by dietary sorbose in nonobese diabetic mice.

The effect of dietary sorbose on the prevention of the incidence of diabetes in the nonobese diabetic mouse was investigated in animals from 5 to 11 wk of age. When sucrose (200 g/kg diet) in the control diet was replaced by sorbose, body weight was significantly reduced. The blood glucose level also was lowered in mice fed sorbose, but the serum insulin level was unchanged. Glucose was not detected in the urine of mice fed the sorbose diet during the experiment, but some mice in the control group excreted glucose in urine. Relative weights of the heart, liver and left kidney were significantly higher in mice fed the sorbose diet vs. those fed the control diet. The results suggest that dietary sorbose would benefit patients with diabetes by lowering blood glucose and inhibiting urinary glucose excretion.

Administration, Oral

Maternal and fetal atrial natriuretic peptide levels, maternal plasma renin activity, angiotensin II, prostacyclin and thromboxane A2 levels in normal and preeclamptic pregnancies.

To clarify the possible role of elevated atrial natriuretic peptide (ANP) in the pathophysiology of preeclampsia, we measured ANP, renin activity (PRA), angiotensin II (Ang II), TXB2 (a stable metabolite of TXA2) and 6-keto-PGF1 alpha (a stable end product of PGI2) concentrations in the plasma of 19 normal pregnant women and 35 severe preeclamptic patients at term. Plasma ANP levels in the preeclamptic patients (n = 35, 71.5 +/- 3.8 pg/ml, mean +/- S.E.) and also umbilical plasma ANP (n = 35, 83.0 +/- 4.2 pg/ml) were significantly (p less than 0.01) higher than those of normal pregnant women plasma (n = 19, 58.7 +/- 3.7 pg/ml) and umbilical plasma (n = 19, 47.6 +/- 4.7 pg/ml). There was a significant (p less than 0.01) positive correlation between maternal ANP levels and fetal ANP levels (n = 54, r = 0.44). Plasma PRA and 6-keto-PGF1 alpha levels in preeclampsia were significantly (p less than 0.05) lower than those of normal pregnancy. The ratio of 6-keto-PGF1 alpha/TXB2 in preeclampsia was significantly (p less than 0.01) lower than that of normal pregnancy as we reported previously. There was no significant correlation between plasma ANP level and plasma PRA, Ang II, plasma TXB2 and 6-keto-PGF1 alpha concentrations. Moreover there was no significant correlation between plasma ANP level and the severity of preeclampsia. These data suggest the possibility of a transplacental crossing of ANP secreted by feto-placental unit, which might be, at least in part, responsible for the high ANP levels observed in preeclampsia. The ANP in preeclampsia is not related directly to hypertension, but it may play a substantial role in the regulation or normalization of blood volume and vascular reactivity.

6-Ketoprostaglandin F1 alpha

[Studies on the nontuberculous lung mycobacteriosis in Japan. (Report of the study in the year 1987 and 1988 of the Mycobacteriosis Research Group of the Japanese National Chest Hospitals)--pulmonary infection caused by Mycobacterium kansasii has begun to appear in all over Japan including north Japan Hokkaido].

UNLABELLED: In this study, the Mycobacteriosis Research Group of the Japanese National Chest Hospitals (MRG) presents the reports of study years 1987 and 1988. As reported previously**, pulmonary infection caused by Mycobacterium kansasii occurred principally in South-West Japan (prefectures South-West of Tokyo) and did not appear in North Japan. However, this disease appeared in 1987 and 1988 in Hokkaido (Sapporo Hospital). Accordingly, we may say the disease occurs all over Japan. This is a noteworthy finding newly recognized in the study years. The prevalence rate of nontuberculous lung mycobacteriosis was determined as 2.92 or 2.78 in 1987 and as 2.02 or 1.91 in 1988 per 100,000 population per year. The estimated rates based on the ratio of nontuberculous lung mycobacteriosis against active lung tuberculosis and based on the ratio of nontuberculous lung mycobacteriosis against culture-positive lung tuberculosis well agreed with each other. COMMENT: In this country, chest physicians customarily report their cases of nontuberculous mycobacteriosis including lung tuberculosis, because the payment of treatment for patients with tuberculosis is free. Because of this custom, tuberculosis statistics surely contain cases of nontuberculous mycobacteriosis. Caution about this has been paid in calculating the prevalence rate. From the study year 1987, the MRG chairman moved from Michio Tsukamura, The National Chubu Hospital, to Nobuhiko Kita, The National Kinki Chuo Hospital.

Humans

Molecular cloning and characterization of cDNA for the precursor of rat pituitary adenylate cyclase activating polypeptide (PACAP).

We have isolated and cloned a cDNA for rat pituitary adenylate cyclase activating polypeptide (PACAP) precursor protein from the brain. The nucleotide and the deduced amino acid sequences revealed that the rat PACAP cDNA encoded a 175 amino acid protein highly homologous to the ovine and human PACAP precursors. The rat PACAP precursor protein included a putative signal peptide at the amino-terminus, a PACAP-related peptide (PRP) of 29 amino acid residues and the rat PACAP of 38 amino acid residues flanked by basic processing sites, and a carboxy-teminus amidation signal for PACAP. Furthermore, the amino acid sequence of the rat PACAP was completely identical to that of the ovine and human PACAP. Northern blot analysis of rat brain RNA revealed an approximately 3.0 kb transcript for the PACAP precursor in rat brain.

Amino Acid Sequence

Specific expression of human endothelin-2 (ET-2) gene in a renal adenocarcinoma cell line. Molecular cloning of cDNA encoding the precursor of ET-2 and its characterization.

Analysis of culture medium of human renal adenocarcinoma cells ACHN by RP-HPLC suggested that the cells specifically secreted human endothelin-2 (ET-2). cDNAs encoding human ET-2 precursor were cloned from a cDNA library constructed with mRNA derived from the ACHN cells, and the nucleotide and deduced amino acid sequences were determined. The ET-2 cDNA was revealed to contain 1.3 kb and encode prepro-ET-2 protein consisting of 178 amino acid residues. The ET-like sequences found in the prepro-ET-1 and -ET-3 was conserved in this prepro-ET-2. The Northern blot analysis of mRNA suggested that the transcript of ET-2 gene was 1.4 kb. This is the first direct evidence that human ET-2 gene was expressed specifically in tumor cells.

Adenocarcinoma

One of the endothelin gene family, endothelin 3 gene, is expressed in the placenta.

A cDNA encoding human endothelin 3 (ET-3) precursor was cloned from a cDNA library from the placenta, and its nucleotide and deduced amino acid sequences were determined. This ET-3 cDNA was found to contain 2.3 kb pairs and encode prepro-ET-3 protein consisting of 224 amino acid residues. The putative big-ET-3 seems to consist of 42 amino acid residues. Two of the intron insertion sites were determined with information from nucleotide sequences of the cloned genomic ET-3 gene. This is the first direct evidence that the ET-3 gene is transcribed and expressed in the placenta.

Amino Acid Sequence

A novel peptide which stimulates adenylate cyclase: molecular cloning and characterization of the ovine and human cDNAs.

A novel neuropeptide which remarkably stimulates adenylate cyclase in rat anterior pituitary cell cultures has been recently isolated from ovine hypothalami by A. Arimura and his collaborators (Biochem.Biophys.Res.Commun.164, 567-574(1989)). This peptide was designated as PACAP38(Pituitary Adenylate Cyclase Activating Polypeptide with 38 residues). In an attempt to investigate physiological implications of PACAP38, we have succeeded in cloning the cDNAs encoding the precursor of PACAP38 from ovine hypothalamus and human testis. An ovine cDNA encodes a protein of 176 amino acids in which PACAP38 is proceeded by a putative signal peptide and a "pro"-region (107 amino acids), and followed by a Gly-Arg-Arg sequence for proteolytic processing and amidation. Deduced amino-acid sequence of human PACAP38 was completely identical to that of the ovine isolated peptide. Cloning of PACAP38 cDNAs confirms the expression of the corresponding mRNAs and the presence of this neuropeptide in ovine hypothalamus and also in human testis.

Adenylyl Cyclases

Secretion and clinical significance of atrial natriuretic peptide in patients with muscular dystrophy.

The plasma concentration of atrial natriuretic peptide was measured in patients with muscular dystrophies to study its relationship with congestive heart failure. In patients with Duchenne muscular dystrophy, the plasma atrial natriuretic peptide concentration was 35.5 +/- 3.3 pg/mL (mean +/- SE), which was higher than that in age-matched normal subjects (9.8 +/- 0.6 pg/mL). It increased with progression of disability and showed significant correlations with the cardiothoracic ratio and the ratio of the preejection period to the left ventricular ejection time. In patients with other types of muscular dystrophy, the plasma atrial natriuretic peptide concentration showed no significant change. Immunohistochemical examination demonstrated many atrial natriuretic peptide-positive cells in atrial muscle of an autopsied patient, indicating preservation of the peptide until the end stage. These findings suggest that measurement of the plasma atrial natriuretic peptide concentration is useful for evaluating heart failure in patients with Duchenne muscular dystrophy.

Adolescent

Plasma renin activity, angiotensin II, prostacyclin and thromboxane A2 concentrations in 139 preeclamptic patients.

Plasma renin activity, plasma concentrations of angiotensin II (AngII), stable metabolites (6-keto-prostaglandin F1 alpha: 6-keto-PGF1 alpha) of prostacyclin (PGI2) and a metabolite (thromboxane B2: TXB2) of thromboxane A2 (TXA2) were measured with radioimmunoassay(RIA) in 107 normal pregnancy (control) and 139 preeclamptic patients in 28-41 gestational weeks. PRA and 6-keto-PGF1 alpha were significantly higher and AngII was slightly higher in preeclampsia than in control, and TXB2 was significantly lower in preeclampsia in control. The ratio of 6-keto-PGF1 alpha/TXB2 was significantly lower in preeclampsia than in control. These data suggest that the changes in the renin-angiotensin system may not be primary alterations in preeclampsia. It can be speculated that in preeclampsia the changes in absolute concentrations of 6-keto-PGF1 alpha and TXB2 are less important than the decrease in the ratio of the 6-keto-PGF1 alpha/TXB2.

Adult

Synthesis of human endothelin-1 precursors in Escherichia coli.

Endothelin, the most potent vasoconstrictor found in nature, is thought to be important in the regulation of blood pressure and/or local blood distribution. Human placenta cDNA fragment encoding preproendothelin-1 (preproET-1) and its carboxyl terminal mature precursor (C-matured precursor) was expressed in E. coli. These products were characterized by both enzyme immunoassay and Western blot analysis.

Blotting, Western