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Biomedical subjects

C Castilla

Publications and source records attributed to C Castilla.

At least 19 recordsLinked to original sources

Immunological properties of a DNA plasmid encoding a chimeric protein of herpes simplex virus type 2 glycoprotein B and glycoprotein D.

A DNA plasmid containing a chimeric sequence encoding both herpes simplex virus type 2 (HSV-2) glycoprotein B (gB) and glycoprotein D (gD) external domains (pcgDB) was used to immunize BALB/c mice against genital HSV-2 infection. To determine the efficacy of this vaccine, groups of mice immunized with the pcgDB plasmid were compared with animals immunized with plasmids corresponding to the individual proteins (pcgBt or pcgDt), administered separately or in combination (pcgBt + pcgDt). We studied the response of the different mouse groups to viral challenge by analyzing clinical disease (vaginitis), serum antibody levels, as well as lymphoproliferative responses and cytokine production by spleen cells. Increased IFN-gamma levels correlated with prolonged survival in mice immunized with the plasmid pcgDB, relative to mice immunized with plasmids coding for the individual proteins alone or in combination. Our results show that immunization with the plasmid encoding the chimeric protein is advantageous over separate proteins. These findings may have important implications for the development of multivalent DNA vaccines against HSV and other complex pathogens.

Animals↗

Smoke impacts from agricultural burning in a rural Brazilian town.

Agricultural and silvicultural biomass burning is practiced in many undeveloped portions of the Amazon basin. In Rond nia, Brazil, such burning is restricted to a brief period in the dry season of August and September to minimize the duration of air quality impacts and to attempt to control escaped fires. During this period, much of the region and the communities within it experience significant exposure to smoke from agricultural and forest fires. In cooperation with Brazilian scientists of the University of Brasilia, the Brazilian Organization for Agricultural Research (EMBRAPA), and the Alternative to Slash and Burn Program coordinated by the International Center for Research in Agroforestry (ICRAF), ambient air quality was measured in Theobroma, a small town in Rond nia, during one week of the open burning period of 1995 to supplement available air quality data and to foster public awareness of the impacts of widespread fires. Personal sampling equipment was used to measure ambient levels of formaldehyde (HCHO), acrolein, CO, benzene, and respirable PM in outdoor air. The data obtained were compared with established Brazilian and U.S. ambient air quality guidelines. Ambient levels of respirable PM averaged 191 microg/m3, HCHO averaged 12.8 ppb, CO averaged 4.2 ppm, and benzene averaged 3.2 ppb. Almost all acrolein samples were less than the detection limit of 1 ppb. The results showed that the public can be exposed to relatively high levels of pollutants under the prescribed burning smoke management strategy of a two- to three-week prescription burning period, although this is an improvement over past years when burning was unregulated and continued through most of the dry season. The results also demonstrate the feasibility of using personal exposure monitoring equipment for low-cost surveys of ambient air quality in polluted regions.

Agriculture↗

[Effect of losartan on human platelet activation by thromboxane A2].

INTRODUCTION AND OBJECTIVES: Previous studies have demonstrated that losartan, an AT-1 receptor antagonist of angiotensin II (Ang II) could block the receptor of thromboxane A2 (TXA2) in the vascular wall. The aim of the present study was to assess the effect of losartan on human platelet activation. MATERIALS AND METHODS: Platelets were obtained from 15 healthy men between the age 26 and 40. Platelet activation was measured by changes in the light transmission of platelet-rich plasma stimulated by a synthetic TXA2 analogue, U46619 (5 x 10(-6) mol/l). RESULTS: The U46619-stimulated platelet aggregation was significantly inhibited by losartan in a dose-response manner. Only a high dose of EXP 3174 (5 10-5 mol/l), the in vivo active metabolite of losartan, was able to attenuate U46619-induced platelet activation. Captopril, an angiotensin I-converting inhibitor failed to modify U46619-induced platelet aggregation. Despite the platelets expressing AT-1 type receptors, of Ang II exogenous Ang II did not modify platelet aggregation induced by U46619. The binding of U46619 to platelets was competitively inhibited by losartan in dose-dependent manner. However, only a high dose of EXP 3174 reduced the binding of U46619. Captopril failed to modify the binding of U46619 to platelets. CONCLUSIONS: Losartan decreased platelet aggregation by a TXA2-dependent mechanism. EXP 3174 showed a lesser potency than losartan to reduce TXA2-platelet activation. Captopril and exogenous angiotensin II had no effect on human platelet activation. These results suggest that losartan reduced TXA2-dependent platelet activation independently of the blockade of AT-1 receptors.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

Doxazosin modifies Bcl-2 and Bax protein expression in the left ventricle of spontaneously hypertensive rats.

BACKGROUND: Increased apoptosis has recently been reported in the heart of spontaneously hypertensive rats (SHRs). OBJECTIVE: To investigate the molecular basis of apoptosis in the left ventricle of SHRs in terms of the expression of Bcl-2 protein (which protects from apoptosis) and Bax protein (which acts as an apoptotic promoter). In addition, we analysed the involvement of alpha -adrenergic receptors in the left ventricular apoptosis of SHRs. METHODS: The study was performed in untreated SHRs (n=16) and SHRs that were orally treated with doxazosin (10 mg/kg body weight per day, for 15 days), a selective alpha1-receptor blocker (n=16). A group of Wistar-Kyoto (WKY) rats (n=16) was used as the control. RESULTS: The left ventricles of untreated SHRs showed a significant increase in Bcl-2 protein expression and a reduced presence of Bax protein. The ratio of Bcl-2:Bax in SHRs was higher than in WKY rats, suggesting an anti-apoptotic state. Paradoxically, both the number of apoptotic cardiac cells and the cleavage of an 85-kDa fragment of the poly (ADP-ribose) polymerase (PARP), a marker of caspase-3 activity, were higher in the left ventricle of SHRs than in WKY rats, suggesting an apoptotic situation. Bax promotes cell apoptosis when it is bound to Bcl-2. We then determined the abundance of Bax-Bcl-2 complexes in the left ventricle of the two groups of animals. Bax-Bcl-2 complexes were more abundant in SHRs than WKY rats. In a second set of experiments, we analysed the role of alpha1-adrenergic blockade by doxazosin in the above-described mechanisms. Doxazosin treatment reduced the formation of Bax-Bcl-2 complexes in the left ventricle of SHRs, and this was accompanied by a decrease in the levels of 85kDa PARP and a reduction in apoptotic left ventricular cells. CONCLUSIONS: The present work suggests that the presence of Bax-Bcl-2 complexes in the left ventricle could be a more reliable marker of the apoptotic state than the determination of the absolute expression of Bcl-2 and Bax proteins. Moreover, the inhibition of alpha1 -adrenergic receptors by doxazosin decreased the abundance of BaxBcl-2 complexes and promoted a reduction of apoptosis in the left ventricle of SHRs.

Animals↗

Detection of Candida albicans in blood by PCR in a rabbit animal model of disseminated candidiasis.

A model of acute disseminated Candida albicans infection in New Zealand rabbits was developed to determine the sensitivity and accuracy of polymerase chain reaction (PCR) assay compared with the lysis-centrifugation blood culture method. Primers used amplify a DNA fragment from the multicopy gene coding for the small subunit rRNA, highly conserved in fungi. The sensitivity of PCR achieved in rabbit blood samples spiked with Candida albicans was 10-50 CFU/100 microL. A nested-PCR increased the limit of detection 10-fold. The sensitivity achieved exclusively with the lysis-centrifugation method (37.5%) was higher than that obtained with PCR (25%), but lower than nested PCR (52.5%). The combination of both techniques, lysis-centrifugation and nested PCR, increased the overall sensitivity rate to 62.5%. These results have demonstrated that, globally, the nested PCR was more sensitive than both single PCR and lysis-centrifugation culture in detecting C. albicans in blood from immunecompetent rabbits with acute disseminated candidosis. PCR could be a useful complementary technique to traditional methods in the early diagnosis of candidemia.

Animals↗

Renal catabolism of truncated glucagon-like peptide 1.

We have searched for the contribution of the kidney to the catabolism of glucagon-like peptide-1 (7-36)amide or tGLP-1 by analyzing the disappearance of the [125I]tGLP-1 both in vivo, from the plasma of bilaterally nephrectomized (BNX), ureteral-ligated (BUL) and normal rats and in vitro from the perfusate of an isolated rat kidney system. Also, we have measured the degradation of the peptide by the isolated renal tubules. Results from in vivo studies demonstrated that the disappearance half-time (t1/2) of [125I]tGLP-1 was significantly lower in the control than in BUL or BNX rats with the metabolic clearance rate (MCR) being higher in the control than in BUL and BNX group; no difference was found for both parameters between BUL and BNX rats. The urinary excretion of the peptide was negligible. The data from the isolated kidney experiments showed a disappearance of the peptide, which was not due to its spontaneous degradation nor to enzymes released from the kidney to the perfusate. Degradation of the peptide also occurred in the presence of isolated tubules. It was dependent upon the concentration of tubules. This could possibly be due to the action of the brush border-associated peptidases. In conclusion, our results demonstrate that, in the rat, the kidney removes the exogenous tGLP-1 from the peripheral circulation, by a mechanism that involves glomerular filtration and tubular catabolism.

Animals↗

Improving medical students' attitudes toward and skills with the elderly.

This study examined the effect of an experimental program in gerontology and geriatrics on the knowledge, attitudes, and interview skill behaviors of third-year medical students. Experimental group members (n = 45; controls n = 51) participated in four 90-minute group sessions that emphasized psychological, sociodemographic, and physiological aspects of aging and interpersonal communication skills. Program participants developed more positive attitudes and demonstrated more socially skilled behavior during an interview with an older adult.

Adult↗

Treatment of encrusted cystitis caused by Corynebacterium group D2 with norfloxacin, ciprofloxacin, and teicoplanin in an experimental model in rats.

Oral doses of norfloxacin (80 mg/kg of body weight per day) and ciprofloxacin (25 and 80 mg/kg/day) and intramuscular doses of teicoplanin (5 mg/kg/day), all administered once a day for 10 days, were evaluated as a means of preventing encrusted cystitis caused by Corynebacterium group D2. Zinc disks dipped into a 24-h broth culture of these microorganisms were inserted into the bladders of female Wistar rats, and treatment was started 14 days after bacterial challenge. The appearance of encrusted cystitis was directly related to a documented urinary tract infection by these coryneforms (71.7 and 0% for rats with positive and negative urine cultures, respectively). All rats that died between days 18 to 43 after bacterial challenge presented very severe encrusted cystitis, which was prevented by teicoplanin and high doses of ciprofloxacin. Rats surviving up to day 44 after bacterial challenge were sacrificed; they presented a lower incidence of encrusted cystitis which was also less severe, with teicoplanin and a high dose of ciprofloxacin being more active in reducing the rate of positive cultures (78.8 and 65.7% reduction, respectively). All antibiotics and doses used were active in vivo at preventing encrusted cystitis by Corynebacterium group D2, but the best therapeutic effect was obtained with teicoplanin.

Animals↗

Renal catabolism of human glucagon-like peptides 1 and 2.

The renal catabolism of [125I]glucagon-like peptide 1 (GLP-1) and [125I]glucagon-like peptide 2 (GLP-2) has been studied both in vivo, by the disappearance of these peptides from the plasma of bilaterally nephrectomized (BNX), ureteral-ligated (BUL) or normal rats, and in vitro, analyzing their catabolism by the isolated, perfused rat kidney. Results from in vivo studies demonstrated that half-disappearance time for both peptides was lower in controls than in BUL rats, and this value in BUL rats was not significantly different from that in BNX rats. In addition, metabolic clearance rate of GLP-1 was higher in control rats than in the other two groups of animals. Urinary clearance rate of both peptides was negligible. In isolated kidney experiments, values for organ clearance of both [125I]GLP-1 and [125I]GLP-2 were similar to those of inulin clearance, which represents the glomerular filtration rate. Urinary clearance of trichloroacetic acid precipitable radioactivity represented less than 1% of total clearance. In conclusion, these results demonstrate a significant role for the kidney in the plasma removal of [125I]GLP-1 and [125I]GLP-2 by a mechanism that involves glomerular filtration and tubular catabolism.

Animals↗

Effect of Bacteroides fragilis on mortality induced by Escherichia coli in an experimental infection treated with cefotaxime, aztreonam or gentamicin.

The possibility that beta-lactamase-producing strains of Bacteroides fragilis can protect Escherichia coli from cefotaxime was studied in an in-vivo model of peritoneal infection in rats. The protective effect of cefotaxime, aztreonam and gentamicin in peritonitis induced by E. coli alone or combined with B. fragilis was evaluated by analysing mortality at 24 and 48 h after bacterial inoculation and treating the animals with two doses of each antibiotic. Comparisons, by drugs, at 24 and 48 h revealed that a statistically significant high mortality rate was obtained at 48 h when mixed infections were treated with cefotaxime, a drug very active in the infection caused by E. coli alone. Infections by mixed flora or E. coli alone treated with aztreonam or gentamicin did not show any significant difference in mortality rate analysed at 24 or 48 h. These in-vivo results confirm previous in-vitro studies and suggest that cefotaxime could be inactivated in mixed infections if a beta-lactamase-producing strain, such as B. fragilis, is involved in a clinical infection.

Animals↗

In vivo significance of the inoculum effect of antibiotics on Escherichia coli.

The minimum dosage of antibiotics which reduced mortality in rats intraperitoneally inoculated with an Escherichia coli isolate was determined. Low mortality rates (0-10%) were obtained when antibiotics with minimal or no inoculum effect (cefoxitin, cefmetazole and gentamicin) were administered to yield serum levels 3 to 20 times the MIC, while antibiotics with a pronounced inoculum effect (cefotaxime and aztreonam) had to be administered to yield serum levels 200 to 1,000 times the MIC determined with a standard (low) inoculum. Thus, it seems that the inoculum effect observed in vitro with some antibiotics for Escherichia coli may have clinical significance.

Animals↗

Tolerance to bilateral subcutaneous access nephrostomy in dogs.

Subcutaneous nephrostomy is a technique which allows access to the renal pelvis for physiological study in the experimental animal over a prolonged period of time without the complications attendant upon an externally draining appliance. One end of an 8 Fr. polyvinyl tube was placed in the renal pelvis at open surgery and the other end was capped and buried in the subcutaneous space for subsequent access by percutaneous needle puncture. This procedure is preferable to surgically created vesical exstrophy, exteriorization of the renal pelvis or conventional nephrostomy, techniques which have been proposed for such purposes. Tolerance to subcutaneous access nephrostomy was investigated in a group of 10 dogs followed for periods ranging from six months to two years. It was found that bilateral subcutaneous access nephrostomy did not alter kidney function but was associated with altered peristalsis. It was also associated frequently with bacteriuria, but not pyelonephritis. Neither calcific deposits nor calculi were seen as a consequence of the tube, and the procedure was well tolerated by the animal although, in some, progressive weight loss was observed. In general, this technique proved to be an excellent procedure for prolonged physiological studies of the urinary tract in dogs, and might similarly find some clinical application in humans.

Animals↗

[Value of the endoscopic sphinctero-papillotomy by surgical radiocholangiodebitometry].

First results of a prospective study about behavior evaluation of endoscopic sphincterotomy of the ampulla of Vater (EPE) through a serial surgical debitometric (RCDM) assessment are analyzed. Five patients submitted to an EPE looking for a biliary tract decompression because of clinical reasons, prior their surgical cholecystectomy, are comparatively evaluated by serial RCDM with regard to 24 patients receiving a broad transduodenal sphincteroplasty (EPTD). Normal debitometric values (N greater than or equal to 13 cc/min.) were obtained from a sample of 673 surgically explored patients. Debitometric behavior of EPE were comparable in all cases and over our standard (mean = 18.6 +/- 2.3 cc/min.); they showed no differences to surgical EPTD patients (x = 18.3 +/- 2.0 cc/min.). Despite uniform results in the EPE group, a more extensive sample is necessary for definitive conclusions; serial surgical RCDM in patients who received EPE, in spite of limited indications, seems to be a valuable method to analyze early functional events after the procedure.

Adult↗

[Perfusion of the dog tibia isolated for the study of mineral metabolism].

A method for perfusion of isolated canine tibia is described. With this procedure the influence of different albumins on the uptake of tritiated 25-hydroxyvitamin D3 (3H-25(OH)D3) by the tibia has been studied. Bovine serum albumin at 1% concentration seems to be optimum for this system. This experimental approach seems to be useful for the study of mineral metabolism.

Animals↗

In vitro and in vivo study of stone formation by Corynebacterium group D2 (Corynebacterium urealyticum).

Corynebacterium group D2 inoculated into normal human urine formed struvite crystals and an increase in pH and ammonium concentration after 24 h of incubation. Zinc disks dipped into a broth culture of this microorganism and inserted into the bladders of rats produced stones with a mean weight of 12.5 mg (ranging from 1 to 57.7 mg) after 12 days. Analysis of the infrared spectrum determined the stones to be composed of struvite. From these results its seems that stone formation by Corynebacterium group D2 may be possible both in vitro and in vivo, which may confirm a previous report involving these bacteria in human clinical encrusted cystitis.

Animals↗

Alpha- and beta-adrenoceptors in the female dog urethra.

The existence and subtypes of alpha- and beta-adrenoceptors in the female dog urethra were studied in vivo and in vitro by means of agonist and antagonist drugs. Noradrenaline, phenylephrine and B-HT 920, stimulants of alpha, alpha-1 and alpha-2 receptors respectively, caused an increase in the urethra tonicity. Thus indicating that the contractile activity is mediated by alpha-1 and alpha-2 adrenoceptors subtypes. On the other hand, the inhibitory urethral activity is under control of beta-adrenoceptors of beta-2 subtype, since the isoprenaline relaxing action is inhibited when beta-2 receptors are blocked, whilst this effect was not observed when beta-1 receptors were blocked. This fact was proved when beta-2 receptors were stimulated with salbutamol.

Adrenergic alpha-Agonists↗

Adrenergic receptors in the urethra and prostate of the horse.

The presence and types of alpha and beta adrenergic receptors in the urethra and prostate of the horse were studied in vitro using adrenergic agonist and antagonist drugs. The existence of these receptors was shown. This finding was based on the observation that the contractile action was mediated by adrenergic receptors of alpha-1 type, although in the prostate alpha-2 type receptors also participated. Relaxation in both tissues was controlled by receptors of the beta-2 type.

Adrenergic alpha-Agonists↗