High-resolution separation of a PCR product from a mixture of DNA restriction fragments was achieved using capillary gel electrophoresis. The capillary gel electrophoretic separation gives an excellent resolution of two fragments of the 500-bp PCR product and the 506-bp DNA fragment, which differs by only 6 bp, and the complete separation of a broader chain length range of DNA afragments up to 12 kbp within 20 min. The plate number of gel-filled capillary was achieved to be 2 million plates per meter. Capillary gel electrophoresis is applied to the gene diagnosis for heart disease through apolipoprotein E genotyping. The advantages and limitations of capillary gel electrophoresis in the application to PCR analysis and DNA diagnosis are discussed.

Apolipoproteins E↗

PubMedDate not supplied

Effect of ouabain on hemodynamics in acute volume expanded hypertensive dogs.

To evaluate possible roles of endogenous Na+-K+-ATPase inhibitors in vasoconstricted blood pressure elevation produced by acute volume expansion, we administered ouabain (Na+-K+-ATPase inhibitor) intravenously (30 micrograms/kg) for 10 min to dogs, 3 h after volume expansion with dextran in lactated Ringer's solution (20 ml/kg, for 1 h). Acute volume expansion resulted in the elevation of blood pressure associated with an increase in cardiac output. In some dogs the blood pressure remained elevated with gradual increase in total peripheral resistance (Group I) or with sustained high cardiac output (Group II), and in other dogs (Group III) it returned to the control level. Ouabain administration elevated the blood pressure and total peripheral resistance in these groups and sham dogs which did not have volume expansion. And these effects of ouabain were not correlated with the degree of blood pressure or vasoconstriction produced by volume expansion. Thus, it is not likely that endogenous Na+-K+-ATPase inhibitors increased to produce vasoconstricted hypertension after acute volume expansion.

Animals↗

PubMedDate not supplied

Effect of cromakalim (BRL 34915) on hemodynamic and electrocardiographic changes induced by endothelin in dogs.

We evaluated whether cromakalim (BRL 34915), a vasorelaxant agent which acts by opening potassium channels, could affect the systemic effects of endothelin, a newly discovered vasoconstrictive peptide. Intravenous administration of endothelin alone (400 pmol/kg) to anesthetized dogs produced blood pressure elevation, which was associated with an increase in cardiac output in the early phase, and was associated with an increase in total peripheral resistance in the late phase. Electrocardiogram showed significant ST-elevation in II, III, and aVF, and ST-depression in aVR and aVL. The same dose of endothelin given to dogs pretreated with cromakalim did not induce these hemodynamic and electrocardiographic changes. Thus, cromakalim, a potassium activator, inhibited the hemodynamic and electrocardiographic actions of endothelin, suggesting that hyperpolarization due to potassium channel activation inhibited the voltage-dependent calcium channel, which is thought to be a major mechanism for the pressor action of endothelin.

Animals↗

PubMedDate not supplied

Enzyme immunoassay for cortisol in serum using cortisol 21-amine.

Cortisol 21-amine (21-amino-11beta,17-dihydroxy-4-pregnene-3,20-dione) was prepared and an enzyme immunoassay for cortisol in serum was established using cortisol 21-amine conjugated with alkaline phosphatase. The minimal amount of cortisol detected was 1ng/tube and the measurable range was from 1 to 80 microgram/d1, using 10 mu 1 of serum sample. This enzyme immunoassay satisfied the standard criteria of dilution, accuracy and precision. The values correlated well with those obtained by radioimmunoassay. This enzyme immunoassay is applicable to the routine determination of serum cortisol in any clinical laboratory. Cortisol 21-amine was found to be a useful derivative for preparing cortisol-enzyme conjugate in enzyme immunoassay.

Amines↗

PubMedDate not supplied

Relation between low serum cholesteryl-ester transfer activity and abdominal aortic calcification in normolipidemic elderly subjects.

We studied the relation between cholesteryl-ester transfer activity (CETA) and abdominal aortic calcification in elderly subjects. Compared with 10 young healthy subjects (mean +/- S.D. age, 27 +/-2 years) and to 26 elderly subjects without abdominal aortic calcification (79 +/- 7 years), 16 elderly patients with abdominal aortic calcification (82 +/- 6 years) had significantly lower levels of serum CETA. However, there were no differences in the levels of serum lipids and apolipoproteins, including total cholesterol, triglycerides, high density lipoprotein cholesterol, low density lipoprotein cholesterol and apolipoproteins A-I, A-II, B, C-II and E, between the two elderly groups. When the two groups of elderly subjects were considered together, the level of serum CETA did not correlate significantly with any lipids and apolipoproteins. These results provide evidence that CETA may prevent the development of aortic calcification in normolipidemic elderly people.

Journal Article↗

PubMedDate not supplied

Polymorphism of HSP70 gene is not associated with type 1 (insulin-dependent) diabetes mellitus in Japanese.

Heat shock protein (HSP) 70 is one of the stress-induced proteins and a candidate for islet autoantigen of Type 1 (insulin-dependent) diabetes mellitus. Association of PstI-8.5 kb allele of HSP70 gene with Type 1 diabetes was previously reported in the Caucasian population. Since HSP70 gene is located in the class III region of HLA, the association may be due to the linkage disequilibrium between class II HLA and HSP70 genes. To study whether HSP70 gene is associated with Type 1 diabetes independent of class II HLA genes, we analyzed both HSP70 gene and class II HLA genes in 32 Japanese patients with Type 1 diabetes and 31 control subjects. By Southern blot hybridization with restriction enzyme PstI and HSP70 genomic probe, two allelic bands, 9.0 kb and 8.5 kb, were detected as reported in the Caucasian population. The allele frequencies of 8.5 kb in Type 1 diabetic patients and normal controls were 0.39 and 0.44, respectively. There was no significant difference between the two groups. On the other hand, by PCR-RFLP analysis, DQA1*0301, DQB1*0303 and DQB1*0401 alleles were positively associated with Type 1 diabetes and DQA1*01 was negatively associated with Type 1 diabetes. These data suggest that the polymorphism of HSP70 gene was not associated with Type 1 diabetes in the Japanese population, and that association of PstI-8.5 kb allele with Type 1 diabetes observed in Caucasian population appears to be due to the linkage disequilibrium between this allele and HLA-DR3.

Alleles↗

PubMedDate not supplied

Clinical comparison of Alzheimer's disease in pedigrees with the codon 717 Val-->Ile mutation in the amyloid precursor protein gene.

Alzheimer's disease (AD) is the most common cause of dementia (32). Although the majority of cases of AD are sporadic, the most consistent risk factor detected in several epidemiological studies has been a positive family history of the disease (14,21). In addition, many large pedigrees have been described in which AD appears to be inherited as an autosomal dominant disorder. In one such pedigree (F23) a point mutation within the beta-amyloid precursor protein (APP) gene at codon 717 was identified and hypothesized to be pathogenic (10). The mutation results in a valine to isoleucine change in APP (APP717 Val-->Ile). Subsequent screening has revealed four other pedigrees, detailed in this study, in which this mutation co-segregates with AD (13,26,37). In addition, one other pedigree (Tor3) with this mutation has been described (15) and detailed clinical, neuropsychological, and neuropathological data are reported. Tor3 is discussed below in comparison to the findings in the families in this study. The five families we report with the mutation were identified in Britain (1 family), the United States (1 family), and Japan (3 families). The mutation has not been reported in the general population of any of these countries (3,13,26,33). On this basis alone it seems this mutation is pathogenic. Other APP codon 717 mutations have been identified which co-segregate with the disease (4,25). Also, a double mutation in APP at codons 670/671 has been shown to cosegregate with the disease in two large Swedish pedigrees (22). In all cases, there is complete co-segregation of the APP mutation with early onset AD, providing overwhelming statistical evidence that these mutations are pathogenic. We present the clinical features and limited neuropathology of AD in these families with the APP 717 Val-->Ile mutation.

Adult↗

PubMedDate not supplied

Prevention of cardiac allograft arteriopathy by antisense Cdc2 kinase oligonucleotide.

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Animals↗

PubMedDate not supplied

Antisense Cdc2 kinase oligonucleotide inhibits adhesion molecule expression in cardiac allograft arteriopathy.

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Animals↗

PubMedDate not supplied

Diagnosis of gastric adenoma versus early gastric cancer by bromodeoxyuridine immunohistochemistry from gastric biopsy specimen.

Differentiation between gastric adenomas and type IIa early gastric cancer by macroscopic and histologic examinations is often difficult. This study has shown that there are differences in the distribution of S-phase cells in these two types of tumour, and these differences can be exploited to differentiate between them. The distribution of S-phase cells was assessed by Bromodeoxyuridine (BrdU) immunohistochemistry using an in vitro BrdU-labelling method. Biopsy specimens were obtained from 27 lesions (12 adenomas and 15 type IIa early gastric cancers), and the distributions of BrdU-labelled cells in the tumour thickness as well as in the superficial layer of the tumour were determined. Significant difference was seen in the labelling of the two tumour types with regard to both these parameters. Using the criteria of BrdU immunohistochemistry, the accuracy of differentiation between gastric adenomas and cancers was 88.9%. Therefore, it is concluded in this study that in vitro BrdU labelling is a useful method of diagnosing gastric adenomas and cancers.

Adenoma↗

PubMedDate not supplied

Localization of the histamine H(2) receptor, a target for antiulcer drugs, in gastric parietal cells.

BACKGROUND/AIMS: Histamine H(2) receptor antagonists are widely used for the treatment of peptic ulcer disorders. However, whether the H(2) receptor is present in parietal or immune cells in the lamina propria remains controversial. This study is designed to determine the H(2) receptor localization immunohistochemically using an antibody against the newly cloned mouse histamine H(2) receptor. METHODS: We cloned the mouse histamine H(2) receptor gene and generated a specific antipeptide antibody against the C terminus. Immunohistochemical studies were performed with this antibody and with a monoclonal antibody against H(+)/K(+) adenosine triphosphatase (ATPase). RESULTS: Histamine H(2) receptors were localized on the plasma membrane and on the cytoplasm just beneath the plasma membrane on the basolateral sides of gastric cells. Confocal microscopy of double-stained sections using the monoclonal antibody against H(+)/K(+) ATPase, a specific parietal cell marker, showed that histamine H(2) receptors colocalized with H(+)/K(+) ATPase. No specific histamine H(2) receptor immunoreactivities were observed in the submucosal regions. CONCLUSION: The H(2) receptor is localized in the gastric parietal cell.

Amino Acid Sequence↗

PubMedDate not supplied

Plasma norepinephrine and dietary sodium intake in normal subjects and patients with essential hypertension.

To evaluate the relationship between sodium intake and the activity of the sympathetic nervous system in patients with essential hypertension, plasma catecholamine levels were measured in 49 essential hypertensive patients and 38 age-matched normal subjects under regular-, high-, and low-sodium diets (mean 24-hour sodium excretions; 116 +/- 8, 267 +/- 29, 31 +/- 7 mEq/day, respectively). The levels of plasma norepinephrine were significantly (p less than 0.01) higher in hypertensive patients than in normal subjects. However, they were significantly reduced by high-sodium intake and increased by low-sodium intake in both patients and controls. The percent decrease and change in the absolute plasma norepinephrine levels from low- to high-sodium states were greater in normal subjects than in the hypertensive patients. The results are interpreted as indicating that an abnormal relationship exists between sodium intake and the activity of sympathetic nervous system in patients with essential hypertension.

Adult↗

PubMedDate not supplied

Increased plasma norepinephrine in young patients with essential hypertension under three sodium intakes.

Increased sympathetic nerve activity may play an important role in the pathogenesis of essential hypertension. It is well known that both dietary sodium intake and age influence the plasma norepinephrine (NE) concentration. The present study was undertaken to evaluate the effects of age on sympathetic nerve activity in patients with essential hypertension and normal control subjects under low-, regular-, and high-sodium regimens (mean 24-hour sodium excretions: 30 +/- 4, 116 +/- 7,280 +/- 15 mEq, respectively). Plasma NE and epinephrine (E) were analyzed by trihydroxyindole methods after high-performance liquid chromatography separation. Subjects were categorized by age into young (less than or equal to 40 yrs), middle-aged (40-60 years), and old (greater than or equal to 60 years) subgroups. Mean plasma NE in hypertensive patients was significantly higher (p less than 0.01) than in normal subjects on each of the sodium regimens. In normal control subjects, there was a significant positive correlation between age and plasma NE with all three sodium intakes. However, no correlation was seen in hypertensive patients on any of the sodium regimens, because in the young subgroup of hypertensive patients the mean plasma NE was significantly higher than that of normal control subjects. These results suggest that the increased sympathetic nerve activity plays an important role in the pathogenesis of essential hypertension, especially in young patients.

Adult↗

PubMedDate not supplied

Role of acetyl glyceryl ether phosphorylcholine in blood pressure regulation in rats.

The role of an endogenously occurring acetyl glyceryl ether phosphorylcholine (AGEPC) in blood pressure regulation was studied with an AGEPC antagonist in rats with hypertension of various etiologies. The hypotensive activity of an intravenously injected AGEPC was competitively suppressed by the intravenous infusion of 3-(N-n-octadecylcarbamoyloxy)-2-methoxypropyl-2-thiazolioethylphospha te (CV-3988) and was dose-dependent. The CV-3988 was infused intravenously into one- and two-kidney, one clip hypertensive, deoxycorticosterone-salt hypertensive, adrenal regeneration hypertensive, spontaneously hypertensive, and normotensive control rats. The increase in blood pressure caused by CV-3988 infusion in spontaneously hypertensive and normotensive control rats was significant (p less than 0.01 and p less than 0.001, respectively, at 60 min) compared with that caused by vehicle infusion. The increase was not seen in rats with secondary hypertension. In rats with two-kidney, one clip hypertension, the initial rapid decrease in blood pressure seen after unclipping was significantly (p less than 0.05) inhibited by CV-3988 infusion as compared with that by vehicle infusion. These results suggest that endogenous AGEPC may participate in the blood pressure regulation and pathophysiology of some forms of hypertension in rats.

Angiotensin II↗