Search PubMed⌕ Search

Biomedical subjects

H Yoshida

Publications and source records attributed to H Yoshida.

At least 775 records · Page 43Linked to original sources

Helicobacter pylori specific nested PCR assay for the detection of 23S rRNA mutation associated with clarithromycin resistance.

BACKGROUND: Clarithromycin is one of the most important antibiotics for Helicobacter pylori eradication. However, 5-10% of strains are reported to be resistant. It has been shown that one point mutation in the 23S rRNA gene is associated with resistance to clarithromycin. AIMS: To establish a polymerase chain reaction (PCR) system which amplifies a segment of the 23S rRNA gene containing the mutation points with primers specific for H pylori, so that H pylori infection and the mutation associated with clarithromycin resistance can be examined simultaneously. METHODS: To detect H pylori infection and the mutation simultaneously, primers specific for the H pylori 23S rRNA gene were designed based on sequence conservation among H pylori strains and sequence specificity as compared with other bacteria. DNA from 57 cultured strains and from 39 gastric juice samples was amplified in the seminested 23S rRNA PCR. Clinical applicability was evaluated in 85 patients. RESULTS: DNA samples from 57 cultured strains were all amplified. The novel assay and the urease A PCR agreed in 37/39 gastric juice samples with no false positives. The assay did not amplify the DNA of bacteria other than H pylori. Eight of 85 samples had the mutation before treatment. In clarithromycin based treatment, eradication was achieved in 2/5 (40%) with the mutation and 29/34 (85%) without the mutation. CONCLUSION: The assay using gastric juice is quick (within 12 hours) and non-invasive (endoscopy not required), enabling rapid initiation of appropriate antibiotic treatment.

Anti-Bacterial Agents↗

Detection of Chlamydia trachomatis in vaginal specimens from female commercial sex workers using a new improved enzyme immunoassay.

OBJECTIVE: To evaluate the performance of a new improved enzyme immunoassay (EIA) kit for the detection of Chlamydia trachomatis in vaginal swab and endocervical swab specimens from female commercial sex workers, in comparison with a conventional EIA test and a polymerase chain reaction (PCR) assay. METHODS: A high risk group of 163 female commercial sex workers who visited a sexually transmitted disease (STD) clinic in order to undergo screening for major STDs, including chlamydial infection, were enrolled. A total of four swab specimens, including two vaginal and two endocervical specimens, were collected from each woman by a clinician. To identify C trachomatis, a new improved EIA kit (IDEIA PCE), a conventional EIA kit (IDEIA), and PCR assay (Amplicor) were used. Discrepancies in the results were resolved using supplementary PCR assay. A female patient was considered to be infected with C trachomatis if the IDEIA PCE test and PCR test for both sample sites (endocervical and vaginal) gave positive results. Following resolution of these discrepancies, relative sensitivity and specificity, confidence intervals, and predictive values for each type of specimen by each assay were calculated. RESULTS: Of the 163 women tested, 35 (21.5%) were shown to be infected with C trachomatis. The relative sensitivities in vaginal swab specimens were 88.8%, 68.6%, and 91.4% using IDEIA PCE, IDEIA, and PCR, respectively. The relative specificities in vaginal swab specimens were 99.2%, 99.2%, and 100%, respectively. The relative sensitivities in endocervical swab specimens were 85.7%, 77.1%, and 91.4% with IDEIA PCE, IDEIA, and PCR, respectively. The relative specificities in endocervical swab specimens were all 100%. CONCLUSIONS: The results obtained in this study suggest that the sensitivity and specificity of IDEIA PCE test on vaginal swab and endocervical swab specimens were similar to those of PCR assay on the two types of specimen. It is concluded that IDEIA PCE test on vaginal swab specimens is an acceptable, sensitive, and less invasive approach for the detection of C trachomatis in commercial sex workers with a high prevalence of C trachomatis infection.

Adult↗

Minimally oxidized low-density lipoprotein increases expression of scavenger receptor A, CD36, and macrosialin in resident mouse peritoneal macrophages.

Fully oxidized LDL (OxLDL) is believed to contribute to atherogenesis in part by virtue of uptake into macrophages via specific scavenger receptors. This phenomenon results in the formation of cholesterol-loaded foam cells, a major component of atherosclerotic lesions. The present study is directed at examining the effects of OxLDL and minimally oxidized LDL (MM-LDL) on scavenger receptor expression and activity in mouse peritoneal resident macrophages. Macrophages were preincubated with MM-LDL or OxLDL at concentrations of 25 or 50 microg/mL for 24 to 48 hours, after which their ability to bind and take up 125I-OxLDL or 125I-acetylated LDL (AcLDL) was determined. MM-LDL pretreatment induced a clear increase of cell association and degradation of 125I-OxLDL and 125I-AcLDL. Pretreatment with OxLDL also enhanced scavenger receptor activity, but to a lesser degree. Neither native LDL nor AcLDL had any effect. Scatchard analysis showed that preincubation with 50 microg/mL MM-LDL for 48 hours increased the Bmax of 125I-OxLDL and 125I-AcLDL by 139% and 154%, respectively, without significantly changing their affinity. Lipids extracted from MM-LDL also significantly induced scavenger receptor activity, but to a lesser extent than did intact MM-LDL. MM-LDL pretreatment increased both mRNA levels and protein levels of scavenger receptor A, CD36, and macrosialin. On the other hand, OxLDL pretreatment increased expression of macrosialin only. These results, showing that MM-LDL can upregulate scavenger receptor expression in mouse resident peritoneal macrophages, suggest that clearance of OxLDL by macrophages in lesions is more effective, in part because the OxLDL precursor, MM-LDL, primes the macrophage for foam cell generation.

Animals↗

Relationship between endothelial function and fibrinolysis in early hypertension.

Abnormalities in fibrinolysis, endothelial function, and glucose and lipid metabolism have been reported in hypertension. This study was conducted to examine the interrelationships between fibrinolytic factors, glucose and lipid metabolism, and endothelial function in hypertension. The effects of administering an angiotensin converting enzyme inhibitor, benazepril, were also examined. Blood levels of the following substances were measured in patients with borderline and mild hypertension (n=50, 51+/-19 years) and in age-matched controls (n=10): total cholesterol, triglycerides, tissue plasminogen activator activity and antigen, and plasminogen activator inhibitor type 1 activity and antigen. Insulin sensitivity was assessed by oral glucose tolerance test, and endothelial function was assessed by evaluating changes in diameter of the brachial artery during reactive hyperemia as observed by ultrasonography. Activities of tissue plasminogen activator and plasminogen activator inhibitor type 1 were both elevated in the hypertensive patients. Stepwise multiple regression analysis showed that plasminogen activator inhibitor type 1 antigen correlated with insulin sensitivity, total cholesterol levels, and triglycerides levels (P<.01). Endothelial function was negatively correlated with tissue plasminogen activator activity and antigen (P<.01). The chronic administration of benazepril (5-10 mg/d) for 20 weeks improved insulin sensitivity, endothelial function (6.6+/-3.4-->9.0+/-2.5%, P<.01), and tissue plasminogen activator activity and antigen. These results indicate that abnormalities in fibrinolysis are associated with endothelial dysfunction as well as disorders of glucose and lipid metabolism in patients with borderline and mild hypertension. The treatment of such patients with benazepril appeared to improve the impairment in fibrinolysis and endothelial dysfunction.

Adult↗

Endothelin-1 production by human synoviocytes.

Immunoreactive (ir)-endothelin (ET)-1 concentrations in serum samples and synovial fluids from patients with rheumatoid arthritis were higher than concentrations in sera obtained from healthy volunteers. No significant difference in ir-ET-1 concentrations in synovial fluid was observed between rheumatoid arthritis patients and osteoarthritis patients. Cultured fluids of synovial cells collected from synovial tissues and leucocytes from synovial fluids of rheumatoid arthritis patients were studied to determine the origin of ir-ET-1 in synovial fluids. Ir-ET-1 was detected in the cultured fluids of synovial macrophage-like type A cells, but not in those of fibroblast-like type B cells from the synovial tissues or leucocytes from the synovial fluids. Longitudinal studies showed that the ir-ET-1 concentration in the cultured fluid reached a peak around 24 h after starting the culture. ET-1 secreted from macrophage-like synoviocytes may be involved in the pathogenesis of inflammatory arthritis.

Arthritis, Rheumatoid↗

Isolation and properties of a novel phospholipase A from rat brain that hydrolyses fatty acids at sn-1 and sn-2 positions.

A Ca(2+)-independent phospholipase A that releases various fatty acids from sn-1 and sn-2 positions was partially purified from rat brain soluble fraction. The enzyme showed an approximate molecular mass of 300 kDa on gel filtration column chromatography. Its enzymatic properties are distinct from those of well characterized phospholipase A2 enzymes; by using a series of synthetic phosphatidylcholines, the enzyme cleaved oleic, linoleic, and arachidonic acids like phospholipase A2, and released palmitic and stearic acids like phospholipase A1. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidic acid were hydrolysed with almost equal efficiencies by this enzyme. These results indicate that the enzyme isolated is a novel Ca(2+)-independent intracellular phospholipase A that might be responsible for production of various fatty acids from membrane phospholipids.

Animals↗

Transgene expression of steel factor in the basal layer of epidermis promotes survival, proliferation, differentiation and migration of melanocyte precursors.

Mutations at the murine dominant white spotting (KitW) and steel (MgfSl) loci, encoding c-Kit receptor kinase and its ligand respectively, exert developmental defects on hematopoietic cells, melanocytes, germ cells and interstitial cells of Cajal. The expression patterns of steel factor (SLF) observed in the skin and gonads suggest that SLF mediates a migratory or a chemotactic signal for c-Kit-expressing stem cells (melanocyte precursors and primordial germ cells). By targeting expression of SLF to epidermal keratinocytes in mice, we observed extended distribution of melanocytes in a number of sites including oral epithelium and footpads where neither melanocytes nor their precursors are normally detected. In addition, enlarged pigmented spots of KitW and other spotting mutant mice were observed in the presence of the SLF transgene. These results provide direct evidence that SLF stimulates migration of melanocytes in vivo. We also present data suggesting that SLF does not simply support survival and proliferation of melanocytes but also promotes differentiation of these cells. Unexpectedly, melanocyte stem cells independent of the c-Kit signal were maintained in the skin of the SLF transgenic mice. After the elimination of c-Kit-dependent melanoblasts by function-blocking anti-c-Kit antibody, these stem cells continued to proliferate and differentiate into mature melanocytes. These melanoblasts are able to migrate to cover most of the epidermis after several months. The SLF transgenic mice described in this report will be useful in the study of melanocyte biology.

Animals↗

A quicker preparation method for autologous fibrin glue.

To establish a quicker preparation procedure for cryoprecipitate (Cryo) from a patient's autologous plasma, to be used as fibrin glue, we examined the effects of various conditions on the concentrations and yields of coagulation factors in Cryo. Human plasma from healthy volunteers was divided and treated under various freezing, shaking and defrosting conditions. The concentrations of fibrinogen, plasminogen, fibronectin, and factor XIII in Cryo were then measured. Results were as follows: (1) concentrations and yields of plasma components in Cryo obtained from plasma stored at -20 degrees C were significantly higher than those in Cryo from plasma stored at -80 degrees C; (2) shaking at 70 cycles/min during the freezing process had a favorable effect on the concentrations and yields of coagulation factors in the Cryo; (3) a shaking thaw process in a cold water bath was a rapid method for obtaining adequate yields of coagulation factors; (4) shaking in the defrosting process did not affect the yields of coagulation factors. These results indicated that Cryo containing high concentrations of coagulation factors could be prepared easily and rapidly from a patient's autologous plasma (within 4-5 h).

Blood Coagulation Factors↗

Urinary metabolites of valproic acid in epileptic patients.

The urinary excretion of valproic acid (VPA) and its metabolites (3-keto VPA, 3-OH VPA, and VPA-glucuronide) in 6 epileptic patients was studied using gas chromatography-mass spectrometry. The amount of VPA and 3-OH VPA excreted in the urine was low (0.1-0.5% of the dose of VPA and 0.6-1.5% of the dose of 3-OH administered). The amount of 3-keto VPA and glucuronide (VPA-Glu) excreted was marked (5.8-26.2% and 13.1-88.7% of the dose of VPA administered, respectively). The urinary excretion of VPA and its metabolites by patients who have taken a normal amount of a VPA preparation was almost the same as that of healthy volunteers. Two epileptic patients who took a large amount of the VPA preparation showed a high excretion of VPA-Glu without an increase in their plasma VPA-Glu.

Adolescent↗

Competitive inhibition of delta8-tetrahydrocannabinol and its active metabolites for cannabinoid receptor binding.

In vitro binding characteristics of delta8-tetrahydrocannabinol (delta8-THC) and its metabolites, 11-hydroxy-delta8-THC (11-OH-delta8-THC) and 11-oxo-delta8-THC, as well as an inactive metabolite, delta8-THC-11-oic acid, as a cannabinoid receptor site from bovine cortex were examined using the specific agonist [3H]CP-55940. 11-OH-delta8-THC and 11-oxo-delta8-THC strongly inhibited the specific binding of [3H]CP-55940. The Ki values of 11-OH-delta8-THC and 11-oxo-delta8-THC for the specific binding of [3H]CP-55940 were 52 and 143 nM, respectively, whereas that of delta8-THC-11-oic acid was 917 nM. Scatchard plot analyses indicated that 11-OH-delta8-THC and 11-oxo-delta8-THC caused a significant increase in the apparent KD value without changing the apparent Bmax. These results reveal that active metabolites of delta8-THC also competitively bind to the cannabinoid receptor as agonists.

Animals↗

Synthesis and pharmacological activities in mice of halogenated delta 9-tetrahydrocannabinol derivatives.

Seven halogenated derivatives of delta 9-tetrahydrocannabinol (delta 9-THC, 1) substituted on the aromatic ring at the 2 and/or 4 position, 2 (4)-fluoro- (2), 2,4-difluoro- (3), 2-chloro- (4), 2-bromo- (5), 2,4-dibromo- (6), 2-iodo- (7) and 2,4-diiodo-delta 9-THC (8) were synthesized and pharmacological effects such as catalepsy, anticonvulsant effects, hypothermia, pentobarbital-induced sleep prolongation and locomotor activity evaluated by intracerebroventricular (i.c.v., 25 micrograms/head) and intravenous (i.v., 5 or 10 mg/kg) injections in mice. The cataleptogenic effects of 2 and 5 were about three-quarters and two-thirds, respectively, compared to those of 1 (i.v.), though other derivatives were much less active (i.c.v. and i.v.). 2 (for clonic seizures) exhibited a significant prolongation of seizure latency induced by pentylenetetrazol (i.v.). Hypothermic effects of monohalogenated derivatives were comparable to 1 when administered by i.v. injection, whereas the effects of dihalogenated derivatives of 1 were attenuated. In contrast, 3 and 8 exhibited a significant hyperthermic effect in mice. In synergy with pentobarbital, 4 and 5 exhibited a significant prolongation of sleeping time by 1.6- and 1.8-fold, respectively, compared with control (32.4 +/- 2.5 min), although other derivatives did not affect significantly the sleeping time (i.c.v.). However, by i.v. injection, 2, 4, 5 and 7 significantly prolonged pentobarbital-induced sleeping time and reduced locomotor activity. The sleep prolonging effects of 2, 4 and 7 (10 mg/kg, i.v.) were as potent as that of 1 (5 mg/kg, i.v.). 5 and 7 were the most potent derivatives among the synthetic cannabinoids examined in the present study. These results indicate that halogenation of 1 leads to modification of the pharmacological profile of THC.

Animals↗

Comparison of filled-in myocardial segments after early and late reinjection of thallium-201--influence of the timing of reinjection on fill-in.

Thallium-201 (201Tl) late reinjection after stress-redistribution imaging improves the detection of viable myocardium. Recently, early reinjection of 201Tl immediately after stress imaging was proposed as a new method for distinguishing ischemic myocardium, hibernating myocardium, and myocardial scar. However, there are no data on the influence of the timing of reinjection on "fill-in." This study was designed to assess whether the reinjection time influences "fill-in" in chronic coronary artery disease. Thirty-three patients with chronic coronary artery disease were studied. All patients underwent exercise 201Tl tomography. Immediately after stress imaging, 37 MBq of thallium was reinjected earlier than usual and early reinjection delayed image (ERDI) was acquired 3 h later. With the same protocol, all patients also underwent a second study involving late reinjection of 201Tl within 1 week. An additional 37 MBq of thallium was reinjected 3 h after stress imaging, and late reinjection delayed image (LRDI) was obtained 10 min later. All images were analyzed qualitatively using a 4-point grading uptake score. Of the 72 hypoperfused segments on stress images, 66 segments showed fill-in and 6 showed persistent defects on ERDI, and of the same 72 segments 55 segments displayed fill-in and the remaining 17 showed persistent defects on LRDI (p<0.05). The delta uptake score (the uptake score of the delayed image minus that of stress image) in early reinjection was 1.60+/-0.80, which was significantly higher than that in late reinjection (1.24+/-0.94, p<0.01). A small dose of thallium reinjected immediately after stress imaging with delayed images obtained 3 h later is convenient and might provide another technique for determining myocardial viability.

Aged↗

Relationship between calcium channel antagonists and nocturnal hypotension and autonomic imbalance in patients with a previous myocardial infarction.

The present study was conducted to evaluate the effect of calcium channel antagonists on diurnal changes in blood pressure and on autonomic function in 71 patients who were receiving a calcium channel antagonist because they had previously suffered a myocardial infarction. Ambulatory blood pressures and Holter ECGs were recorded simultaneously for 24 h. Autonomic function was assessed by heart rate variability. Nocturnal systolic pressure was > or = 90 mmHg in 63 patients (group I) and < 90 mmHg in 8 patients (group II). Significant day to night changes in high-frequency power (from 4.3 +/- 1.2 to 4.5 +/- 1.0/m2) as well as in the ratio of low-frequency power to high-frequency power (from 1.3 +/- 0.1 to 1.1 +/- 0.2) were observed in group I. whereas such changes were blunted in group II. When the calcium antagonist was discontinued or the dose was reduced in group II, the autonomic imbalance improved along with elevation of nocturnal systolic blood pressure. Thus, nocturnal blood pressure should be monitored when such drugs are administered for the treatment of ischemic heart disease to a patient with a previous myocardial infarction. If nocturnal hypotension occurs, the dose should be reduced or the drug should be discontinued.

Adult↗

Hormonal and cardiorenal responses to acute saline loading in mild congestive heart failure--the effect of angiotensin converting enzyme inhibition.

We investigated cardiorenal responses to acute saline loading in patients with mild congestive heart failure (CHF) with and without angiotensin-converting enzyme (ACE) inhibition. Saline loading (infusion of 250 ml of normal saline) was performed on 10 patients with mild CHF and 10 control subjects. Although saline loading reduced plasma renin activity and plasma angiotensin II to a similar extent in both groups, it increased cardiac output, renal blood flow, and the ratio of renal blood flow to cardiac output in the mild CHF group but not in the control group. After saline loading, urinary sodium excretion was higher in the control than in the mild CHF group. In the mild CHF group, saline loading was performed again after an ACE inhibitor (delapril) had been administered for 5 days. Although delapril increased cardiac output and renal blood flow under basal conditions, saline loading did not affect these variables. Delapril improved urinary sodium excretion after saline loading. Thus, the renin-angiotensin-aldosterone axis may contribute to cardiorenal hemodynamics and renal sodium handling in patients with mild CHF. This effect may attenuate the natriuretic response to acute saline loading, and administration of an ACE inhibitor improves this attenuation.

Adult↗

Cardioprotective effects of an angiotensin-converting-enzyme inhibitor, imidapril, and Ca2+ channel antagonist, amlodipine, in spontaneously hypertensive rats at established stage of hypertension.

The present study was performed to compare cardioprotective effects of an angiotensin-converting-enzyme inhibitor, imidapril, and of a Ca2+ channel antagonist, amlodipine, against the cardiac hypertrophy in male spontaneously hypertensive rats (SHRs) at the established stage of hypertension. Fifteen-week-old SHRs were given imidapril (2 and 5 mg/kg/day) or amlodipine (10 mg/kg/day) by gavage for 8 weeks. Three hours after the 1st treatment, imidapril moderately reduced blood pressure without changing heart rate, while amlodipine caused a marked reduction in blood pressure accompanied by transient tachycardia. At the end of the treatments, ventricular weight in the imidapril-treated groups was markedly lower, but that in the amlodipine-treated group was only slightly lower than that in the vehicle-treated group. Myocardial collagen content in the imidapril-treated group tended to be decreased, and significant reduction was observed in the low-dose group. In another experiment, the isolated heart of the imidapril-treated animals demonstrated better cardiac compliance than that in the vehicle-treated animals. In contrast, amlodipine failed to improve cardiac function. The present results suggest that imidapril possesses advantageous effects to prevent cardiac hypertrophy and deteriorated cardiac function in SHRs of established stage of hypertension as compared with amlodipine.

Amlodipine↗

Clinical study of the effectiveness of a dual amplified immunoassay (IDEIA PCE Chlamydia) for the diagnosis of male urethritis.

A clinical study of patients with male urethritis (n=316) was undertaken to determine the sensitivity potential for a new dual amplified immunoassay (IDEIA PCE Chlamydia). Increased sensitivity (98.8%, 84/85) was obtained for IDEIA PCE Chlamydia compared to a conventional antigen detection test (IDEIA Chlamydia, 81.2%, 69/85) when testing urine samples. In a smaller patient population (n=104) the positivity rate for the first-void urine tested with IDEIA PCE Chlamydia of 30.8% (32/104) was similar to the 27.9% (29/104) obtained from urethral swabs tested with a DNA probe assay (PACE 2). The increased sensitivity of the test was confirmed with a commercial PCR kit (Amplicor) and nested PCR. The IDEIA PCE Chlamydia kit has the sensitivity potential to be a clinically reliable alternative for detecting Chlamydia trachomatis.

Chlamydia Infections↗

Antimicrobial activity of a compound isolated from an oil-macerated garlic extract.

A compound showing antimicrobial activity was isolated from an oil-macerated garlic extract by silica gel column chromatography and preparative TLC. On basis of the results of NMR and MS analyses, it was identified as Z-4,5,9-trithiadeca-1,6-diene-9-oxide (Z-10-devinylajoene; Z-10-DA). Z-10-DA exhibited a broad spectrum of antimicrobial activity against such microorganisms as gram-positive and gram-negative bacteria and yeasts. The antimicrobial activity of Z-10-DA was comparable to that of Z-ajoene, but was superior to that of E-ajoene. Z-10-DA and Z-ajoene are different in respect of substitution of the allyl group by the methyl group flanking a sulfinyl group. This result suggests that substitution by the methyl group would also be effective for the inhibition of microbial growth.

Anti-Bacterial Agents↗

Endogenous immunoreactive ouabain-like and digoxin-like factors in reduced renal mass hypertensive rats.

We evaluated the urinary excretion of immunoreactive endogenous ouabain-like factor (OLF) and digoxin-like factor (DLF) to investigate their pathophysiological roles in sodium metabolism and blood pressure in 5/6-reduced renal mass rats, a model of volume-expanded hypertension. About five-sixths of the kidney mass (5/6 RRM, n = 9) was removed from male Sprague-Dawley rats, or the rats were sham operated (control, n = 10). Both groups were fed regular diets with tap water for 1 wk as a control period, followed by 1% saline solution for 4 wk. Systolic blood pressure (SBP), urine volume (UV), urinary sodium excretion (UNaV), DLF, and OLF were measured on the last 2 d of every week throughout the experimental period. SBP and UNaV were significantly higher in 5/6 RRM rats than in control rats. Urinary DLF significantly increased, reaching peak value in the first week, while OLF increased continuously, reaching peak value in the fourth week. In the first week, there were a significant positive correlations between the change in DLF and the changes in UNaV and SBP. However, the change in OLF was not correlated with changes in either UNaV or SBP. Both SBP and UNaV showed a significant positive correlation with OLF (p<0.001, r=0.547, p<0.001, r=0.658, respectively), whereas DLF significantly correlated with UNaV (p< 0.001, r= 0.584) but not with SBP in 5/6 RRM. These findings suggest that endogenous OLF and DLF coexist in rat urine and that an increased level of OLF, but not DLF, may contribute to the development and maintenance of hypertension. DLF may contribute to renal sodium excretion in this volume-expanded hypertensive rat model.

Animals↗