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Biomedical subjects

M Rodríguez

Publications and source records attributed to M Rodríguez.

At least 19 recordsLinked to original sources
PubMed1998

The bovine IFN-omega 1 is biologically active and secreted at high levels in the yeast Pichia pastoris.

The gene coding for bIFN-omega 1 was isolated from bovine genomic DNA by polymerase chain reaction (PCR). Recombinant bIFN-omega 1 was expressed in the yeast Pichia pastoris and high levels of the recombinant protein (0.4 mg ml-1) were secreted to the culture media. The obtained bIFN-omega 1 showed a cross-species antiviral activity on four mammalian cell lines of calf, pig, hamster and human origin, but this activity was absent on Madin-Darby canine kidney (MDCK) cells. A delivery carrier was developed to permit a better release of bIFN-omega 1. When compared with a control group, an increase in 6 days in the corpus luteum lifespan was obtained in cyclic ewes following three interferon (IFN) intrauterine administrations on days 9, 10 and 11 post-estrus. In summary, these results demonstrated for the first time that biologically active recombinant bIFN-omega 1 was highly secreted by P. pastoris showing antiviral activity in different cell lines and an antiluteolytic effect in cyclic ewes, with no detrimental effects on the animals.

Amino Acid Sequence↗
PubMed1998

Diagnosis of herpetic keratoconjunctivitis by nested polymerase chain reaction in human tear film.

A study was performed to evaluate nested PCR (nPCR) versus viral cultures as method and tear film versus corneal scrapings as specimen in the diagnosis of viral keratoconjunctivitis. Tear film specimens were taken from both eyes and corneal scrapings from the affected eye only in 17 patients with suspected viral keratoconjunctivitis. In 15 of the 17 patients the viral agent of the infection could be detected: 11 patients had herpes simplex virus type 1, two varicella-zoster virus, one both herpes simplex virus type 1 and varicella-zoster virus, and one adenovirus. Overall there was no significant difference between the detection rate for corneal scrapings (85%) and tear film (75%). In both types of specimens nPCR showed a higher detection rate than viral cultures (corneal scrapings: 87.5% vs 31.25%; tear film: 75% vs 12.5%; P 0.05). For the diagnosis of keratoconjunctivitis nPCR is superior to viral culture and tear film is an adequate sample that is easier to collect, causing the patient less discomfort.

Adenovirus Infections, Human↗
PubMed1998

The manganese superoxide dismutase from the penicillin producer Penicillium chrysogenum.

The antioxidant enzyme superoxide dismutase has been studied in order to define mechanisms for the influence of oxygen on penicillin production. Manganese-containing SOD activity was purified from penicillin-producing cultures of the filamentous fungus Penicillium chrysogenum and reverse genetics was used to identify full-length cDNA and genomic clones. Sequence analysis revealed a 630-bp ORF containing three exons and two introns with fungal consensus splice-site junctions. The deduced amino-acid sequence (210 amino acids; 23.13 kDa) includes conserved residues required for enzymatic activity and metal binding, and shares significant similarity with Mn- and Fe-containing superoxide dismutases. The sod gene is present as a single copy in the genome of different P. chrysogenum strains and its expression level is not correlated with penicillin-G productivity.

Amino Acid Sequence↗
PubMed1998

Field studies and cost-effectiveness analysis of vaccination with Gavac against the cattle tick Boophilus microplus.

The control of tick infestations and the transmission of tick-borne diseases remains a challenge for the cattle industry in tropical and subtropical areas of the world. Traditional control methods have been only partially successful and the parasites continue to result in significant losses for the cattle industry. Recently, vaccines containing the recombinant Boophilus microplus gut antigen Bm86 have been developed. These vaccines have been shown to control tick infestations in the field. However, extensive field studies investigating the efficacy and cost-effectiveness of vaccination have not been reported and are needed to appraise the effect of this new approach for tick control. Here is reported the results of the application of Gavac in a field trial including more than 260,000 animals in Cuba. In this study the correlation between the antibody response to vaccination and the effect on ticks fertility is determined. Physiological status of the animals was found to affect the primary response to vaccination but not the antibody titers after revaccination. A cost-effectiveness analysis showed a 60% reduction in the number acaricide treatments, together with the control of tick infestations and transmission of babesiosis, which resulted in savings of $23.4 animal-1 year-1. These results clearly demonstrate the advantage of vaccination and support the application of Gavac for tick control.

Animals↗
PubMed1998

Effect of particulation on the immunogenic and protective properties of the recombinant Bm86 antigen expressed in Pichia pastoris.

The recombinant Bm86 tick antigen expressed in Pichia pastoris is obtained in a highly particulated form, as a distinguish feature of this expression system. This particulated protein, the active principle of the recombinant vaccine Gavac against the cattle tick, have shown high immunogenic and protective properties, probably associated with its own characteristics. To evaluate the effects of particulation on the properties of Bm86, three groups of calves were immunized with particulated or non-particulated recombinant Bm86 and the anti-Bm86 antibody response determined. Animals were challenged with a controlled tick infestation and the protective capacities of both proteins assessed. Humoral immune response and protection in cattle vaccinated with the particulated antigen were higher. These experiments suggested that particulation of the Bm86 expressed in P. pastoris is an important feature for the protective properties of the antigen in vaccine preparations.

Animals↗
PubMed1998

Effects of ethylcholine mustard azirinium ion (AF64A) on the choline acetyltransferase and nitric oxide synthase activities in mesopontine cholinergic neurons of the rat.

The choline analogue, ethylcholine mustard azirinium ion (AF64A), has been proposed as a selective neurotoxin that produces degeneration of central cholinergic neurons. However, the mechanisms of action and the specificity or non-specificity of this toxin are still undefined. In this study, we have investigated the effects of AF64A, in comparison with kainic acid, on cholinergic neurons of the mesopontine formation (pedunculopontine and laterodorsal tegmental nuclei), a neuronal population also expressing nitric oxide synthase, the enzyme responsible for the synthesis of nitric oxide. We used choline acetyltransferase immunohistochemistry as a marker of acetylcholine activity, and nitric oxide synthase immunohistochemistry and NADPH-diaphorase histochemistry as markers of nitric oxide synthase activity. Our results show that the injection of low doses of AF64A produces: (1) an area of cavitation in the injection site of pedunculopontine tegmental nucleus (local non-specific effect), and (2) a transient decrease in choline acetyltransferase immunoreactivity in choline acetyltransferase-nitric oxide synthase neurons in both the ipsilateral laterodorsal tegmental nucleus and the perilesional area of the pedunculopontine tegmental nucleus, while their morphology and nitric oxide synthase immunoreactivity remain unaltered (post-diffusion specific effect). These findings indicate that the loss of choline-related enzymatic activity is not necessarily associated with degeneration of cholinergic neurons, and that the recovery of choline acetyltransferase immunoreactivity may arise from neurons whose activity is diminished during the first postinjection weeks. Taking into account that AF64A is a suitable tool to develop a reversible model of neurological disorders related to cholinergic deficit, further efforts should be directed toward elimination of its local non-specific effect.

Animals↗
PubMed1998

The effect of a high phosphorus diet on the parathyroid cell cycle.

High extracellular phosphorus directly increases parathyroid hormone (PTH) secretion and gene transcription. The present study evaluates the effect of high phosphorus diet on the parathyroid cell cycle in rats with normal renal function. Rats were divided into two groups, receiving either a high phosphorus diet (HPD, P=1.2%) or a normal phosphorus diet (NPD, P=0.6%). The dietary calcium content was 0.6% in both diets. Rats were pair fed and sacrificed on days 0, 1, 5, 10 and 15 after initiation of the diet. The parathyroid glands were removed and parathyroid cells dispersed for evaluation of cell cycle and apoptosis by flow cytometry. Serum calcium, phosphorus, PTH and calcitriol were measured. As compared with NPD, the ingestion of a HPD resulted in an increased number of cells in the S phase of the cell cycle from day 1 to 10 (1.2+/-0.09% vs 0.6+/-0.04% for day 1, 1.2+/-0.11% vs 0.6+/-0.06% for day 5 and 1.0+/-0.09% vs 0.5+/-0.04% for day 10, P<0.01). By day 15, the percentage of cells in the S phase in NPD and HPD were not different. In the rats fed the HPD, serum PTH increased significantly from day 5 through 15 (P<0.01). Parathyroid cell apoptosis was minimal and unaffected by the diet. At day 15, the parathyroid gland size in HPD was increased by 27% as compared with NPD (P<0.05). This increase should be attributed to cell proliferation since parathyroid cell size remained unchanged. Serum calcitriol and calcium were not significantly different in the two groups. In HPD, an increase in serum phosphorus was observed only on day 1. The results show that an HPD results in the stimulation of the parathyroid cell cycle independently of changes in calcium and calcitriol.

Animals↗
PubMed1998

Nonbiocompatible hemodialysis membranes induce apoptosis in mononuclear cells: the role of G-proteins.

Mononuclear cells are activated during hemodialysis. In this study, we provide evidence that in vitro culture of mononuclear cells with Cuprophan, a nonbiocompatible hemodialysis membrane, increases the levels of protein phosphorylation in these cells as well as the expression of surface activation molecules. By contrast, culturing of mononuclear cells with AN69, a more biocompatible membrane, did not increase protein phosphorylation levels or expression of surface activation molecules in these cells. In addition, Cuprophan, but not the AN69 membrane, increased the percentage of mononuclear cell death by apoptosis. Inhibition of G-protein-mediated signal transduction decreased the apoptosis of cells cultured with the Cuprophan membrane. The GTP-binding protein involved in Cuprophan-induced apoptosis was sensitive to the ADP-ribosylating pertussis toxin (PTX). The inhibition of GTP-binding protein decreased apoptosis in the early stage of the activation-induced apoptosis, suggesting that G-proteins are implicated in the transmission of apoptosis-inducing signal(s) but do not interfere with the effector signals that mediate the late stages of apoptotic catabolism. Finally, PTX was capable of inhibiting apoptosis without affecting expression of activation molecules; thus, the inhibition of apoptosis by Cuprophan was not due to quenching of the stimulation signals, because monocytes were still able to be activated by Cuprophan despite the action of PTX. The results obtained in this study suggest that cell activation and apoptosis may be mediated by separate intracellular signals.

Acrylic Resins↗
PubMed1998

[Arterioportal fistula and hemobilia in a patient with hepatic transplant].

The case of a 36-year old male liver transplant recipient hospitalized for upper digestive hemorrhage, jaundice and pain in the right hypochondrium is herein reported. Two hepatic biopsies had been performed 60 and 7 days prior to admission. Bleeding was observed to be from the biliary tract by endoscopy and an arterioportal fistula in the right hepatic lobe by echo-doppler and arteriography was seen. Treatment with selective embolization by arteriography was satisfactory with biliary tract drainage not being required. Doppler echography was used to control the evolution of the patient.

Adult↗
PubMed1998

Ambulatory liver biopsy: complications and evolution in 264 cases.

AIM: To determine the safety and acceptance of outpatient liver biopsies. PATIENTS AND METHODS: Data from all liver biopsies were collected in a prospective way over a period of 18 months. Information was gathered on complications, evolution of patients outside the hospital and opinion relating to the test. All patients were required to previously present: platelet count > 60.000/mm3 prothrombin time within 4 seconds of control and absence of ascites or encephalopathy. Criteria for outpatient liver biopsy also included cooperative patient, a partner or friend who stayed with the patient during 12-24 hours following the test and easy access to the hospital. Out of a total of 378 biopsies, 264 (70%) were ambulatory. RESULTS: Five of the 264 outpatients were hospitalized (1.9%), due to a subcapsular hematoma in one case, persistent pain in 3 cases and sever hypotension in the other; all of them evolved favorably in the first 24 hours. Among the inpatients, 2 had complications (1.7%): one subcapsular hematoma resolved without treatment and one abdominal hemorrhage requiring transfusion. Of the ambulatory patients, 46 (18%) presented pain whilst at home, being more frequent in females than in males (30% vs 15%, p = 0.004) and in those who needed more than one attempt to obtain histological material compared with those of a single attempt (33% vs 17%; p = 0.008). Twenty four hours after the test, 83% of the patients had returned to their normal activities. Ninety five percent of the patients questioned considered that the test was not traumatic, and 88% stated a preference for liver biopsy as a day case procedure. CONCLUSIONS: Liver biopsy performed on an ambulatory basis is safe, well tolerated and acceptable by the majority of patients.

Adolescent↗
PubMed1998

[Validation of the imagamma interface for connecting gamma cameras to personal computers].

An electronic interface, imagamma, has been designed in order to link analog gamma cameras to IBM personal computers (PCs). The performance of the interface was assessed for a camera-computer system composed by a GE Maxicamera II-400T gamma camera and an IBM PC, which has a DX 80486 processor, 8 Mb RAM and a suitable acquisition and processing software. Quality control tests for gamma camera-computer systems, established by the International Atomic Energy Agency in the IAEA-TECDOC-602 for < >, were performed. The main performance characteristics of the gamma camera: intrinsic flood field uniformity, intrinsic spatial resolution and spatial linearity were tested as well as the system time performance for dynamic and gated studies. The results obtained for the camera-computer system performance parameters show a good stability. Those results concerning the dynamic characteristics of the interface are between the limits of acceptability, established for each test by the IAEA standards. We conclude that the imagamma interface has suitable performance characteristics to link analog gamma cameras to PC based computer systems in order to be used with clinical application purposes.

Evaluation Studies as Topic↗
PubMed1998

[Effect of enalaprilat on postischemic systolic and diastolic dysfunction (stunned myocardium) on the isolated rabbit heart].

The aim was to determine: 1) whether Enalaprilat (0.08 mg/kg/min) administration: a) before ischemia or b) at the beginning of reperfusion improved the postischemic systolic and diastolic dysfunction ("stunned myocardium") and attenuated the "hyperfunction" phase at the beginning of reperfusion; and 2) whether creatine kinase (CPK), and lactate dehydrogenase (LDH) activities, and lactate release are involved in the protective effects of Enalaprilat. An isolated isovolumic rabbit heart preparation was used as experimental model and subjected to 15 minutes of ischemia followed by 30 minutes of reperfusion, without (group 1), and with Enalaprilat before the ischemia (group 2) and at the beginning of reperfusion (group 3). Left ventricular developed pressure, and end diastolic pressure (diastolic stiffness) were measured and the time constant of isovolumic relaxation (t, Tau) and the ratio between +dP/dtmax and -dP/dtmax were calculated. For the determinations of lactate release and CPK and LDH activities in the perfusate, samples were taken from the coronary effluent for spectrophotometrical measurements. In comparison to the stunned group (group 1) both Enalaprilat preischemia (group 2) and postischemia (group 3) exerted a significant protective effect on the postischemic recovery of contractile state and diastolic stiffness, and attenuated the "hypercontractile" phase in both groups. However Enalaprilat failed to improve myocardial relaxation. Lactate release was also attenuated, but the enzyme activities were not modified.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1998

A recombinant protein-based enzyme immunoassay for IgM antibody to human cytomegalovirus.

The reliability of an enzyme-linked immunosorbent assay (ELISA) which used recombinant antigen to detect human cytomegalovirus IgM was studied. Serum samples from each of 283 children aged 5 +/- 2 years were studied. In all samples the anti-IgM antibodies were investigated with the ELISA techniques Enzygnost (Behring) and ETI-Cito (Sorin) which were both based on whole viral particles, and OPUS (Behring) based on recombinant antigen. Of the samples, 254 (89.4%) were negative with all three tests. The 29 remaining samples were positive with one or two of the three techniques. The diagnostic efficacy of Enzygnost, ETI-Cito and OPUS, respectively, was for sensitivity of 50%, 66.7% and 50%, and for specificity of 100%, 95.6% and 96%, respectively. The results with each of the three ELISAs did not differ widely and their diagnostic efficacy was similar. The method based on recombinant antigen was not found to be more effective than tests based on whole viral particles.

Antibodies, Viral↗
PubMed1998

[Testicular granulocytic sarcoma as a form of relapse in a patient with acute megakaryoblastic leukemia].

Granulocytic sarcoma (GS) is a rare extramedullary tumor composed of myeloblasts and other granulocytic precursors. GS is mostly associated with myeloproliferative disorders, myelodysplastic syndromes and acute myeloid leukaemia. These tumors arise in the absence of leukaemia, at its initial diagnosis or at the time of recurrence. The most common sites of involvement are bone, skin, soft tissue and lymph node. Reports of GS in testis are very rare. We report an unusual case of GS in a patient with megakaryoblastic leukaemia arising in the left testis after four months in complete remission attained with low doses of Ara-C and granulomonocytic stimulating factor.

Aged↗