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L Castillo

Publications and source records attributed to L Castillo.

At least 91 records · Page 5Linked to original sources
PubMed1995

Development of highly specific monoclonal antibodies for the diagnosis of Vibrio cholerae 01.

We report here the development of two monoclonal antibodies, termed 5G8 and 5C12, belonging to the IgM and IgG1 class, respectively, suitable for the identification of Vibrio cholerae 01 in clinical and environmental samples. The specificities of the monoclonals were evaluated by ELISA and indirect immunofluorescent microscopy of microorganisms normally present in stool samples and with two bacterial panels. One panel included 72 potentially antigenically related bacterial strains and the second panel included 20 pathogenic bacterial strains involved in diarrhea cases. The results of these extensive analyses indicate that monoclonal antibodies 5G8 and 5C12 are highly specific and suitable for the clinical diagnosis of Vibrio cholerae 01 in human stool samples by indirect immunofluorescent microscopy. Although the antigenic sites recognized by these antibodies were not identified in this study, the observation of Western blot patterns suggested that 5G8 and 5C12 monoclonal antibodies bind to LPS epitopes, a good structural marker for the detection of V. cholerae 01 because it is present in all bacterial cell walls.

Animals↗
PubMed1995

Phenylalanine and tyrosine kinetics in young men throughout a continuous 24-h period, at a low phenylalanine intake.

We determined the daily rates of whole-body phenylalanine oxidation (phe-Ox) and hydroxylation (phe-OH) in young men receiving [1-13C]phenylalanine and [2H2]tyrosine via primed, constant intravenous (n = 5) or oral (n = 7) infusions for a consecutive 24 h (12-h fast followed by 12-h fed period), and given a low-phenylalanine (21.9 mg.kg-1.d-1), no-tyrosine, but otherwise adequate L-amino acid-based diet for 6 d before the tracer study. Estimates of the daily rates of phe-Ox and phe-OH were significantly higher (P < 0.001) for the subjects receiving the oral tracer, with estimates of phe-Ox obtained with the oral tracer during the 12-h fast period being close to those predicted from similar 24-h leucine kinetic studies (Am J Clin Nutr 1994;59:1000-11). There was generally poor agreement between the measured 24-h rates of phe-Ox and phe-OH compared with the daily rates as predicted from the last hour of the 12-h fast and 5th hour of feeding. From the 24-h data, daily phenylalanine balances were estimated to be positive with the intravenous-tracer protocol and negative with the oral-tracer group. Our results question the adequacy of current international recommendations for aromatic amino acid requirements in healthy adults.

Administration, Oral↗
PubMed1995

Plasma arginine, citrulline, and ornithine kinetics in adults, with observations on nitric oxide synthesis.

The plasma fluxes of ornithine (Orn), arginine (Arg), and citrulline (Cit) and rate of conversion of labeled ornithine-to-citrulline (QOrn-->Cit) were estimated in six healthy adult men receiving an arginine-rich or arginine-free L-amino acid-based diet, each for 6 days. On day 7 an 8-h (3-h fast, 5-h fed) primed continuous intravenous infusion of L-[guanido-15N,15N] arginine, L-[ureido-13C]citrulline, L-[5,5,2H2]ornithine, and L-[5,5,5-2H3]leucine was conducted. Mean citrulline fluxes (mumol.kg-1.h-1) were 10.4-13.6 for the various conditions and/or diets and remained unchanged (P > 0.05). Arginine flux was lowered (P < 0.01) by 38% for fed state during arginine-free period. Ornithine fluxes for arginine-rich were (P < 0.01) reduced with the arginine-free diet. Rates of QOrn-->Cit declined by 30% (P < .05) during the fed arginine-free period. Short-term restriction in the dietary supply of arginine did not alter the rate of whole body nitric oxide synthesis. One subject showed a very high output of nitrate on arginine-free diet (6 times average for remaining subjects).

Adult↗
PubMed1995

Urea cycle intermediate kinetics and nitrate excretion at normal and "therapeutic" intakes of arginine in humans.

We investigated the effects of a high dietary supplement of arginine on plasma arginine, ornithine, and leucine kinetics and on urea production and excretion in five healthy young adult men. Subjects received either 56 or 561 mg arginine.kg-1.day-1 for 6 days via a complete L-amino acid diet, and on day 7 a tracer protocol (first 3 h fasted; next 5 h fed) was conducted, involving primed constant intragastric infusions of L-[15N2-guanidino,5,5-2H2]arginine, L-[5-13C]ornithine, L-[5,5,5-2H3]leucine, and [15N2]urea, with a prime of H13CO3. Plasma arginine and ornithine fluxes increased significantly (P < 0.05) with arginine supplementation, as did the rate of conversion of plasma labeled arginine to ornithine (P < 0.05) and rate of ornithine oxidation (P < 0.001). However, absolute changes in ornithine kinetics were less than those for arginine or those based on changes expected from the change in arginine intake, implying a complex compartmentation in both whole body arginine and ornithine metabolism. The plasma NO3 concentration, daily output of total NO3, and conversion of [15N]arginine to NO3 did not differ between the diets. Urea production and excretion were reduced significantly with arginine supplementation, suggesting an anabolic effect on the whole body nitrogen economy, possibly via the raised plasma insulin levels (P = 0.013) during the prandial phase.

Adult↗
PubMed1995

Inhaled nitric oxide does not alter endotoxin-induced nitric oxide synthase activity during rat lung perfusion.

Nitric oxide (NO) has been demonstrated to decrease its own synthesis in tissue preparations. We tested the hypothesis that endogenous NO synthesis induced by lipopolysaccharides (LPS) would be decreased by exogenous NO during isolated lung perfusion. Rats were pretreated with either saline or LPS 48 h before lung harvest. Endogenous NO synthase activity was measured as conversion of L-[14C]-arginine to L-[14C]citrulline during 90 min of perfusion. NO (100 ppm) was added to the ventilating gas during perfusion of lungs from one group of control or LPS-treated rats. A second group of control or LPS-treated rats was exposed chronically to 100 ppm NO for the 48 h before lung harvest, in addition to receiving 100 ppm NO added to the ventilating gas during lung perfusion. We conclude that conversion of L-[14C]arginine to L-[14C]citrulline was minimal in control lungs and increased in response to LPS pretreatment. NO added to the ventilating gas for the 90 min of ex vivo perfusion did not alter the rate of L-[14C]citrulline production. In vivo exposure to 100 ppm NO for 48 h did not alter the induction of inducible NO synthase activity as measured during ex vivo lung perfusion. This indicates that inhaled NO does not exert negative-feedback inhibition on inducible NO synthase in the ex vivo rat lung.

Administration, Inhalation↗
PubMed1995

Whole body arginine metabolism and nitric oxide synthesis in newborns with persistent pulmonary hypertension.

Despite the potential relevance of the L-arginine-nitric oxide (NO) pathway in the pathophysiology of pulmonary hypertension, no in vivo studies of the kinetics of arginine and NO have been conducted previously in this population. The terminal guanidino N-atom of L-arginine is the precursor for NO, which is oxidized to the stable inorganic nitrogen oxides, nitrite (NO2-) and nitrate (NO3-). Thus, synthesized NO is detected in serum or urine as NO2- and NO3-. The purpose of this investigation was to compare studies of whole body arginine metabolism twice in nine patients with persistent pulmonary hypertension of the newborn (PPHN), using a primed constant i.v. infusion of L-[guanidino-15N2,5,5(2)H2]arginine and L-[5,5,5(2)H3]leucine, first during acute pulmonary vasoconstriction and again during convalescence, and thereby to characterize quantitative aspects of whole body arginine kinetics and NO production, as estimated from the rate of transfer of the 15N-guanidino-label of arginine to urinary nitrate (15NO3-). Arginine flux rates were 84.1 +/- 8.6 mumol.kg-1 h-1 (mean +/- SEM) during acute pulmonary hypertension and increased to 125 +/- 13.2 (p < 0.05) during convalescence, whereas leucine fluxes were unchanged (168.5 +/- 15 versus 178.8 +/- 10.2 mumol.kg-1 h-1), and comparable to those reported in healthy newborns. During convalescence total urinary nitrate excreted increased by 66% (p < 0.05), urinary 15NO3- increased from 0.29 +/- 0.07 to 0.74 +/- 0.15 mumol.d-1 (p < 0.05), and the rate of plasma arginine conversion to NO increased from 10.3 +/- 2.2 to 45.6 +/- 13 mumol.d-1 (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Arginine↗
PubMed1995

[Bacterial translocation in a model of small intestine autotransplant in dogs].

BACKGROUND: Sepsis is the commonest complication of small bowel transplantation. These infections are presumably caused by bacterial translocation, due to splanchnic ischemia. AIM: To study bacterial translocation in the immediate postoperative period after a small bowel transplantation in dogs and to relate it to splanchnic ischemia. METHODS: Three groups of dogs were studied. In group A (n = 6) spontaneous episodes of splanchnic ischemia were monitored in the first 18 h of the postoperative period. In group B (n = 5), a 60 min ischemia was induced by superior mesenteric artery occlusion, two hours after small bowel transplantation. In group C (n = 5) a 60 min ischemia was induced by occlusion of mesenteric vein, two hours after transplantation. Bacterial translocation was assessed through bacterial cultures from the mesenteric vein and splanchnic ischemia with intramucosal pH measurement (a pH < 7.2 was considered indicative of ischemia). RESULTS: Twenty eight of 83 cultures were positive, specially for Gram negative bacilli. The incidence of positive cultures was 14% for group A, 17% for group B and 79% for group C (p < 0.01 compared to groups A and B). The higher incidence of bacterial translocation occurred during the first two hours after transplantation, when the lower intramucosal pH recordings were obtained. The percentage of positive cultures was 39% during periods of ischemia, compared to 24% during periods without ischemia (p = NS). CONCLUSIONS: Bacterial translocation occurs during the first two hours after intestinal transplantation, in concomitance with the lower intramucosal pH readings.

Animals↗
PubMed1994

The plasma flux and oxidation rate of ornithine adaptively decline with restricted arginine intake.

We hypothesized recently that arginine homeostasis is achieved in humans largely by modulating the rate of arginine degradation. We have tested this hypothesis further by measuring in vivo the whole body rate of conversion of arginine to ornithine and ornithine oxidation in six healthy young adults. Subjects received for 6 days an L-amino acid-based diet supplying an arginine-rich or arginine-free intake and on day 7, following an overnight fast, an 8-h tracer protocol (first 3 h, fast state; next 5 h, fed state) was conducted; L-[guanidino-15N2; 5,5-2H]arginine and L-[5-13C]ornithine were given as primed, constant intravenous tracers; measurements of the abundances of various isotopologs of arginine, ornithine, and citrulline in plasma were made, and from these the various kinetic parameters of the metabolism of these amino acids were derived. Arginine and ornithine fluxes were significantly (P < 0.001) reduced in the fed state with arginine-free feeding. The rates of conversion (mumol.kg-1.h-1; mean +/- SD) of plasma arginine to ornithine for arginine-rich were 12.9 +/- 2.6 and 24.7 +/- 4.8 for fast and fed states. These values were 11.1 +/- 3.5 and 9.6 +/- 1.2 (P > 0.05 and P < 0.001), respectively, with an arginine-free diet. [13C]Ornithine oxidation was reduced (P < 0.001) by 46% during the fed state when the arginine-free diet was given. The findings strengthen our hypothesis that homeostasis of arginine metabolism in the human host depends importantly upon a regulation in the rate of arginine degradation with, perhaps, little involvement in the de novo net rate of arginine synthesis.

Adult↗
PubMed1994

Plasma arginine kinetics in adult man: response to an arginine-free diet.

To explore the response of whole-body arginine metabolism to a change in arginine intake, plasma arginine kinetics were investigated in eight healthy adult men who received an L-amino acid diet supplying an Arg-rich or Arg-free intake for 6 days before undergoing a tracer study on day 7. The tracer protocol lasted for 8 hours. For the first 3 hours subjects remained in the postabsorptive (fasted) state, and during the following 5 hours they consumed small meals at 30-minute intervals. Primed continuous intravenous infusions of L-[guanidino-13C]arginine, L-[5,5,5-2H3]leucine, and [15N2]urea were administered to estimate plasma amino acid fluxes and the rate of urea production. For the fasted and fed states, plasma arginine fluxes (mumol.kg-1.h-1, mean +/- SD) were 69 +/- 8 and 87 +/- 12 (P < .01), respectively, for the Arg-rich diet and 63 +/- 14 and 51 +/- 7 (P < .01, from Arg-rich) for the Arg-free diet. Compared with the Arg-rich results, fed-state plasma arginine and ornithine concentrations were decreased (P < .01) and citrulline concentration was increased (P < .01) during the Arg-free diet period. Leucine fluxes and rates of urea production did not differ between the diet groups. The lower fed-state arginine flux in subjects receiving the Arg-free compared with the Arg-rich diet appears to be entirely due to the decreased rate of entry of arginine from the intestine in the former group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Phenylalanine and tyrosine kinetics for different patterns and indispensable amino acid intakes in adult humans.

In a previous paper (Am J Clin Nutr 1993;58:670-83) we described results for plasma amino acid changes, leucine kinetics, and body leucine and nitrogen balance in 20 young men receiving diets for 3 wk. The diets were based on the 1985 FAO/WHO/UNU amino acid requirement pattern (modified FAO diet; n = 7), the Massachusetts Institute of Technology (MIT) requirement pattern (MIT diet; n = 7), or the egg-protein pattern (Egg diet; n = 6). It was concluded, in comparison with the MIT and Egg diets, that the modified FAO diet was not capable of maintaining body amino acid homeostasis or balance. Here we report results from tracer studies with L-[O-2H5]phenylalanine and [2H2]tyrosine that were carried out within the same experiment. The modified FAO diet failed to maintain a mean body phenylalanine balance as determined from rates of phenylalanine hydroxylation (corrected for deuterium-isotopekinetic effects); balance was achieved with the MIT and Egg diets. These results further underscore the inadequacy of the internationally proposed amino acid requirement pattern for healthy adults. We recommend interim use of the MIT, tentative amino acid requirement values in all considerations of adult human amino acid requirements and nutrition.

Adult↗
PubMed1994

[Characterization of a multiresistant strain of Vibrio cholerae O1, isolated from a case of cholera in Chile].

This report characterizes a multiresistant Vibrio Cholerae O1 strain, isolated from a patient with cholera, and investigates the mechanism of resistance. The analyzed strain was resistant to tetracycline, chloramphenicol and trimethoprim-sulfamethoxazole. The resistance was mediated by a 101 megadalton plasmid that was transferred to the resultant of a conjugation assay between the multiresistant V. Cholerae strain and E. coli C-600 used as receptor strain, that acquired the triple resistance of the parental strain. The resistant V. cholerae strain had a Ogawa serotype, El Tor biotype and toxigenic capacity, demonstrated by ELISA and latex agglutination techniques. The biochemical features of the strain were identical to those of susceptible strains, except for the resistance to 10 and 150 ug o 129 vibriostatic factor. The emergence of plasmid mediated resistance to drugs of choice in the treatment of cholera must alert Chilean and Latin American health authorities, considering the cholera will continue affecting the region.

Anti-Bacterial Agents↗
PubMed1994

Estimation of the time required by the malaria parasites to cross the digestive tract to reach blood in mice inoculated by the oral route.

In a previous report we described the transmission of the malaria parasites by the oral route in a murine model. Later, we performed some experiments to demonstrate the transmission of malaria infection by cannibalism. Now we commence to look for the site, mechanism and stages of the parasite involved in crossing the alimentary canal to reach blood and start the infection. To know the invasive stage of the parasite and the way it penetrates, we wanted first to find the level of the digestive tract through which the parasites cross, to restrict the area to be studied. We proposed that the crossing place would be known, if the crossing time of the parasite could be established. Mice were orally inoculated with Plasmodium yoelii yoelii infected blood and their blood was transferred at different times into clean recipient mice intraperitoneally. Malaria infection detected in recipient mice proved that infective forms of the parasite were circulating in the donor mice at the time the blood samples were taken. In this way, we observed that: i) although most parasites required between 2 to 10 min for crossing the alimentary canal, in some case the process can last for 22 hrs; ii) the parasites circulate in blood for variable periods of time (only two minutes in the shortest, and from 10 min on in the longest) being infective to blood recipients. Most orally-inoculated mice whose blood infects other mice, became transient carriers of parasites unable to establish in them.

Administration, Oral↗
PubMed1994

[Characterization of Neisseria meningitidis isolated fron systemic infections. Chile, 1992-1993].

BACKGROUND: in Chile, all systemic infections caused by Neisseria meningitidis must be reported and the bacterial strain must be sent to a Reference Laboratory at the Instituto de Salud Pública de Chile (ISP). AIM: to report the characterization of strains of N. meningitidis isolated during systemic infections in Chile during the years 1992 and 1993. METHODS: the serogroup, serotype, subtype and antimicrobial susceptibility of every strain of N. meningitidis received at the ISP during 1992 and 1993 was studied. RESULTS: six hundred twenty eight strains of N. meningitidis were confirmed during 1992 and 1993. B serogroup was responsible of 91.1% and 94.7% of confirmed cases during 1992 and 1993 respectively. Serotypes and subtypes most frequently associated to B serogroup were B: 15: P1.3 (63.2%) in 1992 and 51.8% in 1993) and B:NT:P1.3 (11.7% in 1992 and 21.3% in 1993). In 1992, all strains were susceptible to penicillin, chloramphenicol, ceftriaxone and rifampicin. During 1993, 7 (2%) strains were found, for the first time in Chile, moderately susceptible to penicillin and rifampicin MIC90 increased fourfold in respect of 1992, although all strains continued to be susceptible to this antimicrobial. CONCLUSIONS: the increasing frequency of NT (non typified strains) isolation will demand the use of molecular biology techniques for their identification. The appearance of penicillin resistant strains in our country is worrisome.

Chile↗
PubMed1994

Phenylalanine and tyrosine kinetics in critically ill children with sepsis.

To better understand the impact of severe illness on the amino acid economy and nutritional needs of pediatric patients, we studied plasma phenylalanine and tyrosine kinetics in eleven critically ill patients (six full-term newborns and five young infants). Within 48 h of the diagnosis of sepsis they were given primed constant i.v. infusions of L-[1-13C]phenylalanine and L-[3,3,2H2]tyrosine for 4 h. Routine nutritional support continued during this period by parenteral administration of dextrose, lipid emulsion, and an amino acid mixture low in tyrosine. Phenylalanine and tyrosine fluxes and rate of phenylalanine hydroxylation did not differ significantly between the two age groups, and so the data were combined for evaluation. For the entire group, values (mumol.kg-1.h-1; mean +/- SD) for phenylalanine and tyrosine fluxes and rate of phenylalanine hydroxylation were 132 +/- 24, 66 +/- 16, and 29 +/- 12, respectively. Plasma phenylalanine to tyrosine concentration ratio was 1.67 +/- 0.6. From a comparison of the rate of phenylalanine hydroxylation with measured phenylalanine intakes, it was concluded that their routine, clinical nutritional support was inadequate to achieve body phenylalanine balance. In comparison with published data, the relative rate of phenylalanine hydroxylation appears to be high. We speculate that tyrosine is a conditionally indispensable amino acid under these conditions; it would be desirable to establish the intake levels and ratio of phenylalanine to tyrosine that effectively support aromatic amino acid balance in these critically ill patients.

Bacterial Infections↗
PubMed1993

Plasma arginine and citrulline kinetics in adults given adequate and arginine-free diets.

The fluxes of arginine and citrulline through plasma and the rate of conversion of labeled citrulline to arginine were estimated in two pilot studies (with a total of six adult subjects) and in a dietary study with five healthy young men. These latter subjects received an L-amino acid-based diet that was arginine-rich or arginine-free each for 6 days prior to conduct, on day 7, of an 8-hr (first 3 hr, fast; final 5 hr, fed) primed continuous intravenous infusion protocol using L-[guanidino-13C]arginine, L-[5,5-2H2]citrulline, and L-[5,5,5-2H3]leucine, as tracers. A pilot study indicated that citrulline flux was about 20% higher (P < 0.05) when determined with [ureido-13C]citrulline compared with [2H2]citrulline, indicating recycling of the latter tracer. Mean citrulline fluxes were about 8-11 mumol.kg-1.hr-1 for the various metabolic/diet groups and did not differ significantly between fast and fed states or arginine-rich and arginine-free periods. Arginine fluxes (mean +/- SD) were 60.2 +/- 5.4 and 73.3 +/- 13.9 mumol.kg-1.hr-1 for fast and fed states during the arginine-rich period, respectively, and were significantly lowered (P < 0.05), by 20-40%, during the arginine-free period, especially for the fed state, where this was due largely to reduced entry of dietary arginine into plasma. The conversion of plasma citrulline to arginine approximated 5.5 mumol.kg-1.hr-1 for the various groups and also was unaffected by arginine intake. Thus, endogenous arginine synthesis is not markedly responsive to acute alterations in arginine intake in healthy adults. We propose that arginine homeostasis is achieved largely via modulating arginine intake and/or the net rate of arginine degradation.

Adult↗