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Biomedical subjects

H Yajima

Publications and source records attributed to H Yajima.

At least 145 records · Page 8Linked to original sources

Detection of human papillomavirus (HPV) type 16 and 52b in cervical cancer tissues by Southern blot hybridization and polymerase chain reaction (PCR).

DNA samples from cancer tissues diagnosed histologically as squamous cell carcinoma of the uterine cervix were examined for the presence of human papillomavirus (HPV) genome DNA by Southern blot hybridization. Of 63 specimens, 32 were found to hybridize with HPV DNA probes; 23 specimens (35%) with HPV type 16 (HPV16), one (2%) with HPV type 18 (HPV18), four (7%) with HPV type 52b (HPV52b), and four others (7%) weakly with HPV52b. Specimens negative for HPV DNA with Southern blot hybridization were subjected to polymerase chain reaction (PCR) to determine the presence of HPV DNA under more sensitive conditions. After PCR using one set of primers specific for HPV16 and HPV52b, 7 out of 31 specimens were found to have HPV16 DNA. None was positive for HPV52b DNA. Our results indicate that HPV52b, as well as HPV16 and HPV18, is associated with squamous cell carcinoma of uterine cervix, and more sensitive determination of HPV infection can be made by amplification of the viral genome by PCR.

Base Sequence↗

Effect of synthetic human cholecystokinin-33 on pancreatic blood flow in dogs.

We have examined the effect of synthetic human cholecystokinin (CCK-33 and CCK-8) on pancreatic blood flow and protein output in anesthetized dogs. Human CCK-33 and CCK-8 increased pancreatic blood flow and protein output in a dose-related manner. There were no significant differences in increasing pancreatic blood flow between human CCK-33 and CCK-8, and increases in blood flow were closely related to the increase of the pancreatic enzyme secretion. L-364,718 (20 nmol/kg) caused a potent inhibition of CCK-stimulated pancreatic blood flow as well as protein output. The degree of inhibition by L-364,718 was dependent on the amount of CCK infused. This study demonstrates that increasing effect on pancreatic blood flow may be one of the biological actions of CCK mediated via CCK receptor. The CCK-33, one of longer molecular forms of CCK, is an important biological stimulator of pancreatic blood flow as well as of exocrine pancreatic secretion.

Animals↗

Enzyme immunoassay of somatostatin (SS)-like immunoreactive substance in bovine milk.

A sensitive and specific enzyme immunoassay (EIA) for somatostatin (SS)-like immunoreactivity (SS-LI) was developed with the use of beta-D-galactosidase labeled antigen. The minimum amount of SS-like immunoreactive substance (SS-IS) detectable by this method was 1.0 fmol/well (25 pmol/l). The level of SS-IS in bovine foremilk was about 20 pmol/l, and the level was unchanged after delivery. On the other hand, the levels of gastrin releasing peptide (GRP)-IS and vasoactive intestinal polypeptide (VIP)-IS in bovine foremilk were very high, but fell during 1 week after delivery to about 10% of those in foremilk.

Animals↗

Micro-enzyme immunoassay of vasoactive intestinal polypeptide (VIP)-like immunoreactive substance in bovine milk.

A sensitive and specific enzyme immunoassay for vasoactive intestinal polypeptide (VIP)-like immunoreactivity was developed with the use of synthetic carboxy-terminal (C-terminal) fragment (residue 11-28) of porcine VIP conjugated with beta-D-galactosidase and a second antibody-coated immunoplate. Using 4-methylumbelliferyl beta-D-galactopyranoside as a fluorogenic substrate, the minimum amount of VIP-like immunoreactive substance (VIP-IS) detectable by this method was 0.1 fmol/well (2.5 pmol/l). The level of VIP-IS in bovine foremilk was above 100 pmol/l, which was more than eightfold higher than that in normal milk.

Animals↗

Detection of human papillomavirus DNA in genital condylomata in women and their male partners by using in situ hybridization with digoxygenin labeled probes.

Twelve couples (12 women and their male partners) presenting genital warts were investigated in order to evaluate the sexual transmission of human papillomavirus (HPV) in mutual partners and the localization of HPV DNA. Formalin-fixed, paraffin-embedded biopsy samples of 12 vulvar condylomata, and 12 penile condylomata from male partners were analyzed for the presence of HPV DNA-6, -11, and 16/18 by using in situ hybridization with digoxygenin labeled DNA probes. HPV DNA was identified in 9 women (75%) and in 9 men (75%). HPV-6 was frequently identified, being revealed in 42% of the vulvar specimens, in 67% of the cervical specimens and 58% of the penile specimens. Seven of 9 (77%) positive couples shared the same HPV DNA, and 2 couples harbored different HPV DNA types between the partners. The signal intensity of the HPV DNA was generally strong in superficial cell layers, weak in parabasal or basal cell layers. No malignant lesions resulted from the condyloma acuminatum caused by HPV-6 or -11. There were only mild dysplasia in the both sexes.

Condylomata Acuminata↗

Isolation and characterization of a tumor-derived human pancreastatin-related protein.

A protein with pancreastatin-like immunoreactivity has been isolated and purified from liver metastasis of a patient with insulinoma. NH2-terminal sequence analysis in conjunction with the use of antibodies specific for the C-terminal structure of pancreastatin identified this protein as a 186-amino acid residue protein corresponding to human chromogranin A-116-301. Using a sensitive radioimmunoassay it was found that serum from the patient with insulinoma contains two peptide species; one comigrates with the 186-amino acid residue pancreastatin and the other the 48-residue pancreastatin.

Amino Acid Sequence↗

[Combination therapy of renal cell carcinoma with interferon-alpha and UFT (or FT-207)].

Human lymphoblastoid interferon (IFN-alpha MOR-22; OIF) was administered to forty-two patients suffering from renal cell carcinoma. Twenty-six patients with metastatic lesions or primary tumor were treated in a clinical trial with either MOR-22 alone or combination of MOR-22 and UFT (or FT-207). The efficacy was assessed in ten of twenty patients who had received MOR-22 alone for more than eight weeks, but objective response was not observed. The efficacy was done in eight of fourteen patients with MOR-22 and UFT (or FT-207) in combination. Complete response was achieved in one, and partial response in two, with an objective response rate of 37.5%. But the difference was no statistically significant in the survival rate of each therapy. In evaluable twenty patients with MOR-22 prophylactically, there was only one case who had occurred lung metastases with a refractory rate of 8.3%. Forty-one patients were examined for side effects to drugs. The main side effects were fever, anorexia, general fatigue, hematologic toxicities, and hepatic dysfunction in both therapy. These side effects occurred with more increased frequency in combination therapy of UFT (or FT-207) compared to MOR-22 alone.

Adult↗

Magainin 1-induced leakage of entrapped calcein out of negatively-charged lipid vesicles.

Effects of magainin 1, a novel antimicrobial peptide, on the permeability of lipid vesicles were investigated by using calcein as a trapped fluorescent marker. Magainin 1 induces the leakage of calcein specifically out of negatively-charged vesicles. The peptide binds to bovine brain phosphatidylserine sonicated vesicles according to the Langmuir isotherm with a binding constant of 3.8.10(5) M-1 and a binding-site number of 0.10 per lipid molecule. The leakage seems to occur at a critical binding number of approx. 0.03 per lipid molecule. A circular dichroism study revealed that magainin 1 conforms mainly to an unordered structure both in an aqueous solution and in the presence of egg yolk phosphatidylcholine vesicles, whereas to an amphiphilic helix with the phosphatidylserine vesicles. In conclusion, magainin 1 interacts with acidic lipids through electrostatic interactions followed by hydrophobic interactions to form an amphiphilic helix, inducing the leakage.

Animals↗

Bioactivity of human pancreastatin and its localization in pancreas.

Synthetic human pancreastatin and its C-terminal fragment were first evaluated with respect to the biological activity on the insulin secretion in the isolated rat islets. Both these pancreastatins inhibited glucose-stimulated insulin secretion at a concentration of 100 nM. The relative molar potency of human pancreastatin compared to that of porcine pancreastatin was equivalent. The pancreastatin-reactive cells were widely located in the islets of Langerhans, and not observed in exocrine acinar cells by immunocytochemistry using human pancreastatin C-terminal specific antibody. These results suggest that human pancreastatin may modulate endocrine function of the pancreas, especially insulin secretion.

Amino Acid Sequence↗

Bioactivity of synthetic C-terminal fragment of rat pancreastatin on endocrine pancreas.

A C-terminal fragment of rat pancreastatin, 26-residue peptide amide was synthesized by the Fmoc-based solid phase method and its biological activity was evaluated for the first time in the conscious rat. Rat pancreastatin inhibited glucose-stimulated insulin secretion and elevated blood glucose levels in a concentration of 10 nmol/kg/h. The relative molar potency of that of porcine is equivalent. This study suggests that the synthetic rat pancreastatin has a biological activity, and may play a physiological role in the endocrine pancreas.

Animals↗

Effects of synthetic human gastrin-releasing peptide on pancreatic exocrine secretion and release of pancreatic polypeptide in conscious rats.

The effects of a newly synthesized peptide, human gastrin-releasing peptide (hGRP), on the pancreatic exocrine secretion and the release of pancreatic polypeptide (PP) were examined in the conscious rat. Plasma PP concentrations were determined by a recently established specific radioimmunoassay for rat PP. Amounts of 0.18, 0.35, and 3.5 nmol/kg/h hGRP significantly stimulated both pancreatic exocrine secretion and 0.35 nmol/kg/h of hGRP increased PP release. Simultaneously infused proglumide (300 mg/kg/h) did not affect either pancreatic exocrine secretion or PP release. However, simultaneous infusion of atropine (100 micrograms/kg/h) slightly inhibited PP release, but did not restrict the incremental response of pancreatic protein secretion to hGRP. These results suggest that hGRP directly stimulates pancreatic exocrine secretion and PP release.

Animals↗

High plasma pancreastatinlike immunoreactivity in a patient with malignant insulinoma.

High levels of pancreastatinlike immunoreactivity were detected in the plasma (2.9 pmol/ml, greater than 200-fold the normal level), pancreas (2.9 nmol/g wet wt, greater than 450-fold the normal level), and liver (1.6 nmol/g wet wt) of a patient with pancreatic insulinoma with metastasis to the liver by a sensitive and specific radioimmunoassay for human pancreastatin. Antiserum was produced against the C-terminal fragment of human pancreastatin-(24-52), which was synthesized according to the sequence of human chromogranin A corresponding to that of pancreastatin. With the antiserum, intense immunocytochemical staining was detected in the tumors. Sephadex G-50 gel filtration showed that the tumors and plasma contained two molecular forms of pancreastatinlike immunoreactivity--a molecular form coeluted with synthetic human pancreastatin-52 and a larger molecular form (Mr approximately 12,000-15,000). The smaller form eluted in the same position as synthetic human pancreastatin-52 on reverse-phase high-performance liquid chromatography.

Adenoma, Islet Cell↗

Effect of synthetic physalaemin on splanchnic circulation in dogs.

Physalaemin has been reported as one of the most potent vasodilator and hypotensive peptides (1-4). In spite of these studies, however, the effect of the peptide on splanchnic circulation is not known precisely. In the present study, the effect of synthetic physalaemin on superior mesenteric arterial blood flow, portal venous blood flow and pancreatic capillary blood flow was investigated in dogs. Dose dependent increases of superior mesenteric arterial blood flow and portal venous blood flow were induced in response to physalaemin (0.1-10.0 ng/kg). Superior mesenteric arterial blood flow and portal venous blood flow attained maximal increases of 77 +/- 8.9% and 70 +/- 8.6%, respectively, at a dose of 5 ng/kg. Physalaemin caused a dose-related decrease in systemic arterial blood pressure. Pancreatic capillary blood flow did not show significant change with the administration of physalaemin. These data suggest that physalaemin may play some physiological roles in the regulation of splanchnic circulation.

Amino Acid Sequence↗

Pedicled and "flow-through" venous flaps: clinical applications.

Recently, the pedicled venous flap and "flow-through" venous flap have been the focus of increasing attention for skin defects of the fingers and hands. For successful venous flap use, the following approaches have been suggested: (1) a pedicled venous flap with preservation of the draining veins alone; (2) a "flow-through" flap with preservation of a flow-through vein in the flap; and (3) an arterialized "flow-through" venous flap which ensures arterial blood flow into the flap. Based on findings that venous blood is helpful in flap survival, the authors made use of the first two flap types, the pedicled venous flap and the "flow-through" venous flap and attempted to establish and clarify reasonable conditions for flap survival. Venous pressure of the finger and elbow was measured and venographies of the finger and hand were also carried out. The following conditions are regarded as essential in successful venous flap procedures: (a) use of a venous flap with a rich venous network; (b) preservation of many "flow-through" veins; (c) harvesting a pedicled venous flap where the veins have afferent (reversed) venous pressure; and (d) anastomosing veins of the "flow-through" flap with recipient veins where high efferent venous pressure exists and differential pressure is observed. Clinical cases are presented and the authors attempt to explain flap failure from previously unknown causes. Conditions for flap harvesting are also discussed.

Adult↗

Effect of L-364,718 on GRP-stimulated pancreatic and gastric secretions and GI peptides in conscious dogs.

The effect of L-364,718, a cholecystokinin (CCK) receptor antagonist, on exocrine pancreatic secretion, gastric secretion, and plasma levels of gastrointestinal (GI) peptides stimulated by gastrin-releasing peptide (GRP) was examined in five conscious dogs. Intravenous infusion of graded doses of synthetic porcine GRP (18, 36, and 178 pmol/kg/h) caused significant and dose-dependent increases in pancreatic and gastric juice secretion and in plasma levels of pancreatic polypeptide (PP), CCK, and gastrin. Intravenous injection of L-364,718 (20 nmol/kg) significantly inhibited GRP-stimulated pancreatic outputs of juice volume, protein, and amylase and plasma PP release. L-364,718, however, did not affect gastric juice volume and plasma levels of CCK and gastrin. The results suggest that endogenously released CCK is, at least in part, responsible for GRP-stimulated pancreatic protein and enzyme secretions and PP release in dogs. The results further suggest that GRP-stimulated pancreatic secretion might be, in part, a direct response of GRP to exocrine pancreas.

Animals↗

Immuno-affinity purification of specific antibodies against human gastrin releasing peptide (h-GRP) by the h-GRP(1-8)-linked polydimethylacrylamide resin.

A new method for immuno-affinity purification of specific antibodies against human gastrin releasing peptide(h-GRP) was developed. The antiserum GP(No. 6201) elicited by h-GRP-BSA conjugate was heterogeneous and reacted not only with h-GRP and its fragments but also partially with other structurally related peptides, such as other GRPs (porcine, canine, and chicken), bombesin, and neuromedin-C. To obtain specific antibodies against human GRP, antiserum GP was purified by column chromatography on the amino-terminal octapeptide h-GRP(1-8)-linked polydimethylacrylamide resin. The antibody thus obtained was highly specific to amino-terminal sequence of h-GRP and hardly reacted with other GRPs (porcine, canine and chicken), bombesin, and even carboxy-terminal h-GRP fragments in ELISA.

Acrylamides↗

Enzyme immunoassay of gastrin releasing peptide (GRP)-like immunoreactivity in milk.

A sensitive and specific enzyme immunoassay (EIA) for gastrin releasing peptide (GRP)-like immunoreactivity was developed using enzyme-labeled antigen. The synthetic carboxy-terminal fragment of human GRP(12-27) was conjugated with beta-D-galactosidase for EIA. The minimum amount of GRP-like immunoreactivity detectable by this method was 0.24 femtomol/well (6 picomol/liter). The level of GRP-like immunoreactive substance in bovine foremilk was about 150 nanomol/liter, the level of which was more than hundredfold higher than that in normal milk or calf serum.

Amino Acid Sequence↗

Identification of a transforming gene of human papillomavirus type 16.

Previously, we observed sequential two-step alteration, growth stimulation, and progression to a more malignant state in NIH 3T3 cells transfected by human papillomavirus type 16 (HPV-16) DNA. In this study, we prepared a cDNA library from RNA extracted from cells transfected with the HPV-16 DNA and isolated cDNA clones which had growth-stimulating activity. Analysis of these cDNA clones indicated that the E7 open reading frame alone is responsible for inducing both steps of this cell transformation.

Animals↗