Biomedical subjects

H Yajima

Publications and source records attributed to H Yajima.

At least 163 records · Page 9Linked to original sources

PubMed1989

Plasma pancreastatin-like immunoreactivity in various diseases.

Plasma pancreastatin (PST)-like immunoreactivity in normal subjects and patients with various diseases was estimated by a RIA, using antiserum raised against a synthetic C-terminal peptide of human PST deduced from the sequence of human chromogranin-A. The mean level +/- SEM was 13.2 +/- 0.6 pmol/L in normal subjects, but was significantly higher in patients with chronic renal failure (526.7 +/- 48.5). An immunoreactive form corresponding to a human PST-like sequence [human chromogranin-A-(250-301)] and a larger form were detected by gel filtration of plasma from these patients, suggesting accumulation of the larger molecular form in these patients. A significant increase in PST-like immunoreactivity was also found in patients with liver cirrhosis (20.8 +/- 3.0 pmol/L), but not in patients with noninsulin-dependent diabetes mellitus, chronic pancreatitis, or pancreatic cancer. Elevated levels were found in 16 of the 21 patients with small cell lung carcinoma examined. High levels were also found in 3 of 11 patients with islet cell tumor.

Adenoma, Islet Cell↗

PubMed1989

Studies on peptides. CLXVII. Solid-phase syntheses and immunological properties of fragment peptides related to human malaria circumsporozoite protein.

A glycine-linked tetramer of Asn-Ala-Asn-Pro, a tandem repeated sequence of malaria circumsporozoite (CS) protein, was synthesized by the Boc-based solid phase method, followed by deprotection with 1 M trimethylsilyl trifluoromethanesulfonate-thioanisole in trifluoroacetic acid. In addition, three tetramer-related peptides were similarly synthesized, i.e., a 34-residue peptide [linked with TH, a proposed T-cell epitope of CS, at the C-terminus of the tetramer], a 46-residue peptide and a 59-residue peptide [linked with HA or HA', two proposed T-cell epitopes of influenza hemagglutinin protein, at the N-terminus of the above 34-residue peptide]. Their immunological properties were examined by enzyme-linked immunosorbent assay, for which three different congenic strains of mouse were used to raise the specific antibodies. Despite conjugation of T-cell epitopes to the tetramer, the mice of low-responder strains to the tetramer failed to produce any antibody specific to the tetramer. However, with the aid of recombinant interleukin 2 as an adjuvant, the low-responder mice produced antibody with relatively high titers.

Antigens, Surface↗

PubMed1989

Protease-catalyzed semisynthesis of human neuropeptide Y.

Human neuropeptide Y was semisynthesized by enzymatic condensation of des-Tyr36-NH2 human neuropeptide Y and H-Tyr-NH2 using Pseudomonas aeruginosa elastase, a metalloenzyme possessing a hydrolytic specificity for the imino side of hydrophobic amino acids. The optimum pH for this enzymatic synthesis was judged to be around 7 in a high concentration of an organic solvent.

Amino Acid Sequence↗

PubMed1989

Immuno-affinity purification of specific antibodies against vasoactive intestinal polypeptide (VIP) on VIP (1-10)-linked polydimethylacrylamide resin.

A novel method for immuno-affinity purification of specific antibodies against amino-terminal (N-terminal) porcine vasoactive intestinal polypeptide (p-VIP) was developed. The antiserum VP (No. 6203) elicited by p-VIP-immunoglobulin G (IgG) conjugate was heterogenous and reacted not only with the N-terminal fragments of p-VIP but also with carboxy-terminal (C-terminal) fragments. To obtain specific antibodies against the N-terminal fragment of p-VIP, antiserum VP was purified by column chromatography on N-terminal decapeptide (p-VIP 1-10)-linked polydimethylacrylamide resin. The antibody thus obtained was highly specific to the N-terminal sequence of p-VIP and hardly reacted with the C-terminal fragments of VIP in enzyme-linked immunosorbent assay.

Antibodies↗

PubMed1989

Detection of human papillomavirus type 52b in cervical dysplastic tissue.

We examined infection in the tissue of cervical dysplasia of human papillomavirus (HPV) 52b, which we recently isolated from cervical cancer. To screen a lot of samples, we used an assay system with an HPV DNA typing kit containing an HPV 33 probe that cross-hybridizes with HPV 52b DNA. DNA samples from 123 patients with cervical dysplasia were first screened with this kit. Sixteen of these appeared to be positive, and they were subjected to Southern blot hybridization with the HPV 52b probe under highly stringent conditions. Of the 16 samples, two were found to have HPV 52b genome DNA. Our results, together with the fact that HPV 52b was isolated from cervical cancer, seem to support the notion that HPV 52b infection is closely associated with the onset of cervical cancer.

Blotting, Southern↗

PubMed1989

[Human papillomavirus DNA in vulvar lesions].

In order to study the role of human papillomavirus (HPV) in genital cancer, we attempt to determine the presence of HPV in tumors of the genital tract. In the current study, we have examined 35 histologically diagnosed cases of vulvar lesions in attempts to compare the sensitivities of various methods of detecting HPV. All but 7 vulvar lesions contained HPV DNA by Southern blot DNA hybridization. Detectability of HPV by immunocytochemistry and koilocytosis significantly declined as compared to DNA hybridization. The detection of HPV DNA by in situ hybridization is discussed.

Blotting, Southern↗

PubMed1989

[Experimental study on secondary living bone graft--method of creating new donor by vascular bundle implantation to isolated bone].

The purpose of this study was to demonstrate the possibility of creating a new donor for a vascularized bone graft by a vascular bundle implantation to an isolated bone. Preliminary experiments on rats were performed preliminarily in order to investigate the revascularization of bone by a vascular bundle implantation and/or by implanting in muscular tissues (experiment-1). When a vascular bundle was implanted to the intramedullary cavity of an isolated bone and then the bone was implanted in muscular tissues, the isolated bone was more readily revascularized and revitalized than under other conditions. In experiment-2, the revascularized bone was transferred to the abdominal subcutaneous space on the vascular pedicle that was an implanted vascular bundle. Histologic and fluorochrome bone labeling studies showed that the revascularized bone was kept revitalized after the transfer. These results demonstrate that muscle and vascular bundle co-operated effectively in the revascularization of bone. Vascular bundle can also act as a pedicle for transferring the revascularized bone.

Animals↗

PubMed1988

Isolation of a new type of human papillomavirus (HPV52b) with a transforming activity from cervical cancer tissue.

Substantial evidence has implicated human papillomaviruses (HPVs) as etiological agents of human cervical cancer. We detected and cloned an unidentified HPV genome in cellular DNA extracted from a surgical specimen of a Japanese patient with cervical cancer. Hybridization studies suggested that this HPV was the most homologous to HPV33 among more than 51 types of HPV ever identified. A recently identified new HPV, isolated by W. Lancaster and designated HPV52, is also most homologous to HPV33 (W. Lancaster et al., unpublished data). Our HPV was found to be homologous but not identical to HPV52 and hence was designated HPV52b. By introducing it into NIH3T3 cells, we found that HPV52b DNA had growth-stimulating activity as HPV16 DNA. This newly identified HPV52b DNA was present as episomes in cervical cancer cells of two out of two specimens so far examined and the integrated copies were not detected. HPV52b was present in three out of 15 (20%) specimens in Japan. In addition, the presence of three additional unidentified HPV sequences homologous to HPV52b was detected in three other specimens. The results suggest that this group of HPV may be prevalent and involved in carcinogenesis of cervical cancer in Japan.

Blotting, Southern↗

PubMed1988

Interaction of human parathyroid hormone-related peptide with parathyroid hormone receptors in clonal rat osteosarcoma cells.

Synthetic peptides corresponding to the amino-terminal region of the human parathyroid hormone-related peptide (hPTHrp) were used to characterize the interaction of hPTHrp with parathyroid hormone (PTH) receptors in clonal rat osteosarcoma cells (ROS 17/2.8). Both hPTHrp-(1-34) and [Tyr40]hPTHrp-(1-40) showed full agonist activity in stimulating cyclic AMP accumulation in ROS cells; human PTHrp-(1-34) was approximately 2.5-fold as potent as hPTH-(1-34). Both [Tyr-40]hPTHrp-(3-40) and hPTH-(3-34) inhibited the cyclic AMP increase induced by either hPTHrp or PTH with parallel dose-inhibition curves. Binding to intact ROS cells of a 125I-labeled [Tyr40]hPTHrp-(1-40) (125I-[Tyr40]hPTHrp-(1-40)) which retains full biological activity was time- and temperature-dependent and reversible. Binding of 125I-[Tyr40]hPTHrp-(1-40) and 125I-labeled [Nle8, Nle18, Tyr34]bovine PTH-(1-34)NH2 to ROS cells was competed for, to the same extent and with the comparable potency, by either unlabeled hPTHrp or PTH peptides. The binding capacity and affinity of receptors in ROS cells were strikingly similar for hPTHrp and PTH. Affinity cross-linking with either radioligand resulted in high affinity, specific labeling of an apparently identical macromolecule centering at Mr = 80,000, which was detected in sodium dodecyl sulfate-polyacrylamide gel electrophoresis in both reducing and nonreducing conditions. The data indicate that hPTHrp and PTH, their amino-terminal fragments at least, interact with the identical receptors with regard to affinity, capacity, specificity, and physicochemical characteristics in osteoblastic ROS 17/2.8 cells.

Animals↗

PubMed1988

Bioactivity of synthetic human pancreastatin on exocrine pancreas.

A biological activities of synthetic human pancreastatin (1-52) and its C-terminal fragment (24-52) were evaluated for the first time in the conscious rats. Both pancreastatins inhibited CCK-stimulated pancreatic secretion in a range of 20-200 pmol/kg/h with the same potency, indicating that the C-terminal portion of this peptide has a full biological activity. The relative molar potency of this substance compared to that of porcine pancreastatin was equivalent. This study suggests that human pancreastatin has the same biological activity as that of porcine, and plays a biological action in the exocrine pancreas.

Animals↗

PubMed1988

Role of sulphated tyrosine residue in influencing the biologic activity of human cholecystokinin-33.

To evaluate the role of the sulphated tyrosine residue in position 27 in human cholecystokinin-33, parallel bioassay of the sulphated form of human cholecystokinin-33 and the unsulphated form of human cholecystokinin-33 was performed on the pancreatic protein secretion. Both peptides increased the protein output in a dose-related manner. However, the sulphated form possessed a considerably higher activity than the sulphated form. The relative potency of the unsulphated human cholecystokinin-33 compared to that of the sulphated human cholecystokinin-33 (taken as 1.0) was 0.08. From the results, it was suggested that the sulphated tyrosine may play an important role in controlling the activity of the longer molecular forms as well as that of the smaller forms of cholecystokinin.

Animals↗

PubMed1988

Effect of synthetic human cholecystokinin-33 on exocrine pancreas.

Synthetic human cholecystokinin-33 was first evaluated with respect to the biological activity on the pancreatic protein secretion. Human cholecystokinin-33 increased pancreatic protein secretion in a dose-related manner. The relative molar potency of this substance compared to that of synthetic cholecystokinin-8 (taken as 1.0) was 0.92. This study supports the concept that longer molecular forms of cholecystokinin are quantitatively important mediators of biological action of cholecystokinin on the pancreas.

Animals↗

PubMed1988

Studies on peptides CLVIII. Model experiments for the synthesis of open-chain unsymmetrical cystine-peptides.

Treatment of a mixture of Cys(R)(O) and Cys(R') with an acid was found to generate cystine in fairly good yields, when suitable R, R', and an acid were selected. An unsymmetrical cystine peptide was prepared by treatment of a mixture of Z(OMe)-Cys(R) (0)-Ala-NH2 (R = Acm or MBzl) and Z(OMe)-Cys(MBzl)-Gly-OBzl with TFA or 1 M TFMSA/TFA3. Oxytocin was obtained in an excellent yield by TFA treatment of the protected peptide containing Cys(Acm)(0) and Cys(MBzl). Thus, formation of the disulfide bond was found feasible at the position of Cys(R) (0).

Cystine↗

PubMed1988

Effect of natural peptide YY on blood flow and exocrine secretion of pancreas in dogs.

Little is known regarding the mechanism by which peptide YY exerts an inhibitory effect on exocrine pancreatic secretion. The purpose of this study is to determine if peptide YY affects pancreatic blood flow with simultaneous measurement of exocrine pancreatic secretion in dogs. Pancreatic blood flow was measured by a laser Doppler flowmeter which allows continuous measurement of tissue blood flow. Natural peptide YY (0.1, 0.5, 1 microgram/kg) was infused intravenously as a bolus under background infusion of secretin (1 unit/kg/hr) in combination with cholecystokinin-octapeptide (0.1 microgram/kg/hr). Peptide YY caused a reduction of pancreatic blood flow in a dose-dependent manner as well as inhibition of pancreatic protein output, attaining the maximal reduction (28 +/- 4%) and inhibition (45 +/- 9%) at a dose of 1 microgram/kg, respectively. Simultaneous and continuous observation on tissue blood flow and exocrine secretion of the pancreas revealed that there was a highly significant correlation between the percent reduction of pancreatic blood flow and that of volume of pancreatic juice in response to peptide YY (r = 0.849, P less than 0.001). This study provides evidence that the mechanism of peptide YY-induced inhibition of exocrine pancreatic secretion is mediated, at least partly, through the decreased pancreatic blood flow.

Animals↗

PubMed1988

Assessment of neuromedin B polyclonal antibodies as molecular probes in neural tissue.

Five unique, high affinity rabbit polyclonal antibodies against neuromedin B were characterized in a radioimmunoassay in terms of the following parameters: pH and type of buffer, ionic strength, and non-ionic detergents in order to optimize immunoglobulin-peptide interaction; specificity using peptides of the bombesin family, in addition to the tachykinin substance P; and affinity to neuromedin B. Optimum conditions included acidic pH (5.25), high ionic strength (greater than 0.1 M) and absence of non-ionic detergents, which inhibited the assay. Affinities for the 5 antibodies ranged from 10 to 48 fmol neuromedin B with titers from 1:1,000 to 1:10,000 and the sequence-specificity covered the entire peptide; cross-reactivity towards substance P was negligible. As a model tissue, rat spinal cord was homogenized with 5 different extraction solvents, including acetone, methanol, acid and alkaline conditions, and assayed by each polyclonal antiserum; neuromedin B immunoreactivity levels were highest in acid and alkaline extracts and reflected the specificity of the antibody used. Applying these antisera to rat brain extracts, the posterior pituitary gland contained the highest concentration of immunoreactive equivalents of neuromedin B followed by the anterior pituitary, hypothalamus, and hippocampus. The immunoreactive content in the pituitary and hypothalamus, however, depended on the particular antisera used with significant (P less than 0.01) differences existing between them. Further application of these polyclonal antibodies to a spinal cord extract analyzed by isocratic reverse-phase HPLC conditions also revealed differences in their cross-reactivity with the immunoreactive peptides. These antisera may now be used as molecular probes for the determination of extractable immunoreactive neuromedin B from neural tissue and in situ localization by immunohistochemical techniques.

Amino Acid Sequence↗

PubMed1988

Gastrin-releasing peptide stimulates cholecystokinin secretion in perfused rat duodenum.

We examined the effect of porcine gastrin-releasing peptide (GRP-27) and other analogous neuropeptides on cholecystokinin (CCK) secretion from the isolated perfused rat duodenum. GRP-27 stimulated CCK secretion in a monophasic pattern and in a dose-dependent manner ranging from 10(-9) M to 10(-6) M, and 10(-7) M of GRP-27 led to an increment of 442 +/- 120.8 fmol/3 min. The stimulatory effect of GRP-27 on CCK was not inhibited by 10(-5) M of atropine. 10(-7) M of neuromedin C and B, analogs of GRP, stimulated CCK secretion to increments of 382 +/- 64.1 and 289 +/- 47.2 fmol/3 min, respectively. Carbachol (10(-9) to 10(-6) M), VIP (10(-9)M), secretin (10(-9)M) and glucose (11 mM) did not stimulate CCK secretion, and the addition of atropine (10(-5)M) to them led to no significant changes. These results suggest that GRP may directly stimulate CCK secretion from the duodenum and work as a non-cholinergic, peptidergic neurotransmitter.

Animals↗

PubMed1988

Effects of synthetic human gastric inhibitory polypeptide on splanchnic circulation in dogs.

Changes in blood flow in the celiac artery, superior mesenteric artery, and pancreas in response to an intravenous injection of synthetic human gastric inhibitory polypeptide (GIP) were determined simultaneously and continuously in anesthetized dogs, using a transit-time ultrasonic flowmeter and a laser-Doppler flowmeter. Injection of GIP significantly increased superior mesenteric arterial flow in a dose-related manner (by 9%, 43%, and 139% at 30 s after an injection at the doses of 3, 50, and 800 pmol/kg, respectively). In contrast, celiac arterial flow was not significantly altered by GIP at any of the three doses. Calculated vascular resistance in the superior mesenteric artery decreased after GIP infusion, whereas that in the celiac artery was not changed by GIP. Pancreatic blood flow decreased significantly after GIP injection at the doses of 50 and 800 pmol/kg (by 11% and 17%, respectively). Our data indicate that there is a substantial difference in the hemodynamic responses to GIP among splanchnic organs, and suggest that GIP acts specifically on the mesenteric vasculature.

Animals↗

PubMed1988

Skin graft including subcutaneous vein: experimental study and clinical applications.

Skin grafting, using full and split thickness techniques, may yield poor results when satisfactory circulation is not restored in the recipient bed. Using a rabbit ear model with a vein located in the center of the graft, donor skin and subcutaneous vein were raised simultaneously, and the vein was preserved, resulting in a viable passage for inflow and outflow. Grafts were shown to survive in this experimental group when circulation was satisfactory after venous preservation. In the control group with no vein preservation, all grafts became necrotic. These results were clinically applied for cases in which skin defects, caused by finger trauma, resulted in poor circulation in the recipient bed. Skin and subcutaneous vein were simultaneously taken from the forearm and grafted successfully, resulting in excellent prognoses.

Adult↗