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Biomedical subjects

R Bermudez

Publications and source records attributed to R Bermudez.

13 recordsLinked to original sources
PubMed2005

Towards single screening tests for brucellosis.

This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.

Animal Diseases↗
PubMed2004

Enzyme immunoassay for the diagnosis of brucellosis: chimeric Protein A-Protein G as a common enzyme labeled detection reagent for sera for different animal species.

A recombinant protein combining the immunoglobulin binding sites of Proteins A and G, conjugated with horseradish peroxidase was used as a universal detection reagent for the assessment of antibodies against Brucella spp. The reagent was applied in an indirect enzyme immunoassay for detection of antibodies to smooth lipopolysaccharide antigen in sera from Brucella spp. exposed and non-exposed cattle, sheep, goats and pigs and to antibodies to rough lipopolysaccharide in sheep, dogs and cattle. The results were similar to those obtained when murine monoclonal antibody-enzyme conjugates were used. An added advantage was that a universal cut-off for all tests using the proteins A and G detection reagent could be established, simplifying diagnostic interpretation of the data.

Animals↗
PubMed2004

Comparison of serological tests for the detection of ovine and caprine antibody to Brucella melitensis.

The indirect enzyme-linked immunosorbent assay (IELISA), the competitive enzyme-linked immunosorbent assay (CELISA) and the fluorescence polarisation assay (FPA) were evaluated with sera from sheep experimentally infected with Brucella melitensis and negative Canadian sheep. The sensitivity and specificity of the assays were as follows: IELISA: 91.7% and 97.6%, CELISA: 75.0% and 99.8% and FPA: 91.7% and 89.5%. Sera from the same experimental population were divided according to serological reaction in the rose bengal agglutination test (RBT) and the complement fixation test (CFT). Reactivity relative to the RBT positive and CFT positive sera were as follows: IELISA: 99.7%, CELISA: 93.2% and FPA: 99.1%. Since sera from goats with proven B. melitensis infection were not available, 699 sera from goats judged positive in the buffered antigen plate agglutination test (BPAT) and CFT and 982 BPAT/CFT negative Canadian goats were used. The sensitivity and specificity of the assays relative to the BPAT and CFT positive sera were: IELISA: 99.4% and 98.0%, CELISA: 95.4% and 97.1% and FPA: 92.7% and 99.8%.

Agglutination Tests↗
PubMed2002

Field trial of the brucellosis fluorescence polarization assay.

Fluorescence polarization assay (FPA) is a homogeneous technique which was applied to the serological diagnosis of bovine brucellosis. Because of its simplicity and because it may be performed very rapidly, it was an ideal test to adapt to field use. The FPA was used to test cattle on six dairy farms in Baja California, Mexico. Anticoagulated blood, serum, and milk were collected from each animal. The anticoagulated blood was tested immediately on the farm while serum and milk were tested subsequently in the laboratory. Cattle on one farm (n = 140) were thought not to be infected with Brucella abortus and the other farms were thought to have high prevalence of the infection. The whole blood FPA (FPA(bld)) did not detect antibody in any of the cattle on the first premise. This finding was confirmed using a number of other serological tests, including the buffered antigen plate agglutination test, the complement fixation test, the indirect and competitive enzyme immunoassays, and the FPA using serum and milk. Cattle on the other premises (n = 1122) were tested in a similar fashion. The sensitivity of the FPA(bld), relative to the serum FPA (considered the definitive test), was 99.1% and the relative specificity of the FPA(bld) was 99.6%. These results compared favourably with those obtained using the other serological tests.

Animals↗
PubMed2001

Growth hormone plus high protein diet promotes adaptation after massive bowel resection in aged rats.

OBJECTIVE: To determine whether GH improves adaptation following massive bowel resection in the aged rat fed on a high protein-content diet. MATERIAL AND METHODS: Seventy-seven male Wistar rats aged 22+/-1 months underwent 80% bowel resection or laparotomy (sham-operation). They were randomly placed into one of eight groups, treated with either growth hormone (1mg/kg/day) or saline, and fed a liquid diet containing either a high or a normal protein content. Intestinal tissue and blood samples were taken seven days after surgery and analysed to measure intestinal mucosal proliferation and mucosal height, as well as plasma levels of IGF-1 and somatostatin. RESULTS: Resection of the small bowel in aged rats remarkably increased villous height and crypt proliferation. Growth hormone did not potentiate the increase in mucosal height and crypt proliferation observed after intestinal resection in aged rats fed a normal protein content diet, but did in those receiving a high-protein diet. Plasma levels of IGF-1 and somatostatin were not modified by surgery or treatment. CONCLUSION: Growth hormone may increase the adaptation of intestinal mucosa in aged rats undergoing massive intestinal resection, but requires an adequate nutritional support with increased amounts of high quality protein.

Adaptation, Physiological↗
PubMed1996

Trimethaphan versus sodium nitroprusside for the control of proximal hypertension during thoracic aortic cross-clamping: the effects on spinal cord ischemia.

Sodium nitroprusside (SNP) has been used to control the proximal hypertension associated with thoracic aortic cross-clamping (TACC) during thoracic aortic surgery. It worsens neurologic outcome, presumably by further decreasing distal arterial pressure and increasing cerebrospinal fluid (CSF) pressure, thereby worsening the spinal cord perfusion pressure (SCPP). Trimethaphan does not increase CSF pressure. Therefore, the present study investigates the effect of trimethaphan versus SNP to control proximal hypertension during TACC on neurologic outcome. Two groups, each with eight mongrel dogs, were studied. All animals underwent descending TACC for 45 min. The mean proximal aortic blood pressure was maintained at 95-100 mm Hg by the use of SNP or trimethaphan. Distal aortic pressure was allowed to vary. The dogs were neurologically evaluated 24 and 48 h later by a blinded observer. During cross-clamping, there was no difference in mean proximal aortic pressure between groups. After 10 min of cross-clamping, distal aortic pressure was higher (P < 0.01), CSF pressure was lower (P < 0.01), and SCPP was higher (P < 0.005) in the trimethaphan group as compared with the SNP group (group effect). Neurologic outcome as assessed by Tarlov's score was better at 24 and 48 h in the trimethaphan group (P < 0.05). Histopathologic injury trended with worsened neurologic outcome. We conclude that 1) trimethaphan produced higher SCPP than SNP, and 2) neurologic outcome was better in the trimethaphan group.

Animals↗
PubMed1990

Ca2+ transport in isolated mitochondrial vesicles from Leishmania braziliensis promastigotes.

Leishmania braziliensis maintained very low (50 +/- 20 nM) intracellular concentrations of calcium ions under normal conditions, as shown by the fluorimetric indicator QUIN2. Digitonin-permeabilized cells liberated large amounts of calcium ions in the presence of the ionophore A23187, indicating the presence of a large intracellular reservoir for this ion. Given the extraordinary extension of the single giant mitochondrion of Kinetoplastida and the known capacity of mitochondria from other sources to accumulate calcium, we tested the capacity of this organelle to accumulate calcium ions in Leishmania. Coupled mitochondrial vesicles, five-fold enriched in succinate-cytochrome c oxidoreductase, were obtained from promastigotes by gentle grinding (45 s) with glass beads in hypertonic buffer solution, followed by differential centrifugation. These vesicles had a respiratory control ratio of 1.82 +/- 0.15, and two phosphorylation sites (sites II and III) using succinate as electron donor, and were capable of calcium uptake in the presence of several respiratory substrates; this uptake was enhanced in the presence of ADP and Pi and was blocked by classical electron transport inhibitors. Uncouplers such as carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP) and the calcium ionophore A23187 released previously accumulated calcium ions, suggesting that the driving force for the calcium uptake by the vesicles is the respiratory generated electrochemical potential gradient of protons. A study of the affinity of this system for calcium showed that even at 90 microM free calcium, succinate-induced calcium uptake is not saturated while approaching a level of 200 nmol min-1 (mg protein)-1, indicating a low-affinity, large-capacity system.(ABSTRACT TRUNCATED AT 250 WORDS)

Aminoquinolines↗
PubMed1988

A technique for donor lung procurement and preservation for transplantation after completion of cardiac donation.

The scarcity of donor lungs for transplantation has been caused, in part, by the belief that a single donor cannot provide usable lungs if it serves as a heart donor. However, the appropriate division of the left atrial wall provides sufficient cuffs for individual transplantation of the heart and each lung into three separate recipients. With the described technique, the results of the present studies demonstrate the feasibility of donor lung procurement and preservation for transplantation after cardiac donation. Use of this method will allow the most effective and efficient use of the limited supply of donor organs and thereby permit therapeutic single lung transplantation in selected patients. Cardiac donation should no longer preclude lung donation as it has in the past.

Animals↗
PubMed1987

Comparative efficacy and safety of ceftizoxime, cefotaxime and latamoxef in the treatment of bacterial pneumonia in high risk patients.

One hundred and thirty-five patients with bacterial pneumonia who had risk factors (alcoholism, chronic obstructive pulmonary disease, corticosteroid therapy diabetes mellitus, advanced age, solid tumours) were randomly allocated in a double-blind fashion to receive either ceftizoxime (2-4 g every 8 h), cefotaxime (1-2 g every 4 h), or latamoxef (2-4 g every 8 h). Of the 84 patients evaluable for efficacy, clinical cure was achieved in 91%, 85%, and 89% of ceftizoxime- (20/22), cefotaxime-(23/27), and latamoxef-treated (31/35) patients, respectively. Adverse reactions occurred in one of 45 ceftizoxime-treated patients, one of 43 cefotaxime-treated patients, and seven of 47 latamoxef-treated patients. Abnormal laboratory values during therapy were seen in 50% of latamoxef-treated and 43% of cefotaxime-treated patients and in 29% of ceftizoxime-treated patients. Hypoprothrombinaemia occurred in five latamoxef-treated patients and one of these patients experienced an episode of haematemesis. In this study, ceftizoxime, cefotaxime, and latamoxef were similarly effective; however, the incidence of side effects was most frequent with latamoxef.

Aged↗
PubMed1985

Characterization of muscarinic receptors in human submandibular salivary glands.

We studied the binding of the muscarinic antagonist [3H]N-methyl-scopolamine [( 3H]NMS) in order to characterize muscarinic receptors located in human submandibular salivary glands obtained from intrasurgical biopsy. [3H]NMS bound with a Kd value of 1.56 nM to a single class of muscarinic receptors (Bmax 37.3 fmol/mg protein) since pirenzepine exhibited a homogeneous binding profile.

Humans↗
PubMed1978

Dengue fever with hemorrhagic manifestations: a report of three cases from Puerto Rico.

During the 1975 dengue epidemic in Puerto Rico, we studied three patients who had serologically confirmed dengue with hemorrhagic manifestations. None of the patients went into shock and none died. Only one of the patients had disease that resembled dengue hemorrhagic fever (DHF) as observed in Southeast Asia. This patient was a 14-year-old boy who had epistaxis, a positive tourniquet test, moderate thrombocytopenia, and significant hemoconcentration. The other two patients had hemorrhagic disease which was of clinical importance, but was not typical of DHF. These cases of dengue with hemorrhagic manifestations are the only ones known to have been documented in Puerto Rico.

Adolescent↗
PubMedDate not supplied

Levamisole activates several innate immune factors in Scophthalmus maximus (L.) Teleostei.

Levamisole, originally synthesized as an anti-helminthic, has been widely used in human and veterinary medicine as an immunomodulator or adjuvant. However, data on its use in fish are scarce, and no information is available for turbot. To study the effects of levamisole treatment on the innate immune system of juvenile turbot, two different doses (D1=500 mg/kg; D2=250 mg/kg dry food) were orally administered for 2 weeks and samplings were performed at -10, 14, 28, 49 and 77 days post treatment (p.t.). Biometrical, haematological, histological and immunological data were obtained. Specific growth rate was higher in the medicated groups than in the control (C), but the difference was statistically significant only for D1 fish at day 49 p.t. The leucocytes/thrombocytes ratio was significantly higher in D1 than in C fish at 14 days p.t., but decreased subsequently. At most samplings, the percentage of the circulating lymphocytes was lower and that of the progranulocytes was higher in the medicated fish than in the C ones. The percentage of fish with high haemotopoietic activity in the kidney was clearly higher in D1 and D2 fish than in C ones at some sampling points. The respiratory burst activity of blood leucocytes was significantly higher in D1 fish than in C ones in all samplings, except at day 77 p.t. when control fish experienced a rebound effect. In all medicated fish, an initial increase of such activity was observed, followed by a further decrease. Their serum peroxidases followed a contrary pattern, with a decrease in the second sampling and a subsequent and non-significant recovery, a situation also observed for serum lysozyme and complement. Therefore, oral levamisole treatment actually affects some turbot immune factors, although stimulation or depression can occur depending on the considered factor and the administered dose. These results point out the interest of further studies on the mechanisms involved in the levamisole action for its adequate use as immunomodulator.

Adjuvants, Immunologic↗