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Biomedical subjects

C Vela

Publications and source records attributed to C Vela.

At least 19 recordsLinked to original sources

[Catastrophic antiphospholipid syndrome after ovulation induction].

INTRODUCTION: The catastrophic antiphospholipid syndrome (CAPS) is a rare expression of the antiphospholipid syndrome (APLS). The all predicting factors of happening of CAPS are not yet discovered. Some women presenting an APLS with infertility have recourse to ovulation induction (OI). EXEGESIS: After an OI, a woman is admitted for acute renal failure, hypertension, seizures, respiratory failure. History and immunologic tests are in favour of APLS. Renal biopsy highlights a thrombotic microangiopathy. Diagnosis of CAPS is made. Patient improves after respiratory assistance, parenteral treatment for hypertension and anticoagulant therapy. CONCLUSION: IO seems to be on of the factors which leads to CAPS. To avoid such consequences, it's essential to know the history and the immunological status of the patients having recourse to this treatment to take care of these risked pregnancies.

Acute Disease↗

Role of galanin and galanin(1-15) on central cardiovascular control.

Galanin and the N-terminal fragment Galanin(1-15) are involved in central cardiovascular regulation. The present paper reviews the recent cardiovascular results obtained by intracisternal injections of Galanin and Galanin(1-15) showing that: (A) the Galanin antagonist M40 blocks the central cardiovascular responses induced by Galanin(1-15) but not those elicited by Galanin; (B) both Galanin and Galanin(1-15) induce the expression of c-Fos in cardiovascular nuclei of the medulla oblongata with different temporal and spatial profiles; (C) the cardiovascular action of Galanin(1-15), but not Galanin, is mediated by peripheral beta-receptor stimulation; (D) and it is demonstrated an antagonistic Galanin/alpha2-adrenoceptors interaction as well as a differential modulation of central cardiovascular responses of Angiotensin II by Galanin or Galanin(1-15). All these data strengthen the involvement of both Galanin molecules as neuromodulators on central cardiovascular regulation.

Animals↗

Large scale production and downstream processing of a recombinant porcine parvovirus vaccine.

Porcine parvovirus (PPV) virus-like particles (VLPs) constitute a potential vaccine for prevention of parvovirus-induced reproductive failure in gilts. Here we report the development of a large scale (25 l) production process for PPV-VLPs with baculovirus-infected insect cells. A low multiplicity of infection (MOI) strategy was efficiently applied avoiding the use of an extra baculovirus expansion step. The optimal harvest time was defined at 120 h post-infection at the MOI used, with the cell concentration at infection being 1.5x10(6) cells/ml. An efficient purification scheme using centrifugation, precipitation and ultrafiltration/diafiltration as stepwise unit operations was developed. The global yield of the downstream process was 68%. Baculovirus inactivation with Triton X-100 was successfully integrated into the purification scheme without an increase in the number of process stages. Immunogenicity of the PPV-VLPs tested in guinea pigs was similar to highly purified reference material produced from cells cultured in the presence of serum-containing medium. These results indicate the feasibility of industrial scale production of PPV-VLPs in the baculovirus system, safety of the product, and the potency of the product for vaccine application.

Animals↗

Inactivated recombinant plant virus protects dogs from a lethal challenge with canine parvovirus.

A vaccine based upon a recombinant plant virus (CPMV-PARVO1), displaying a peptide derived from the VP2 capsid protein of canine parvovirus (CPV), has previously been described. To date, studies with the vaccine have utilized viable plant chimaeric particles (CVPs). In this study, CPMV-PARVO1 was inactivated by UV treatment to remove the possibility of replication of the recombinant plant virus in a plant host after manufacture of the vaccine. We show that the inactivated CVP is able to protect dogs from a lethal challenge with CPV following parenteral immunization with the vaccine. Dogs immunized with the inactivated CPMV-PARVO1 in adjuvant displayed no clinical signs of disease and shedding of CPV in faeces was limited following CPV challenge. All immunized dogs elicited high titres of peptide-specific antibody, which neutralized CPV in vitro. Levels of protection, virus shedding and VP2-specific antibody were comparable to those seen in dogs immunized with the same VP2- peptide coupled to keyhole limpet hemocyanin (KLH). Since plant virus-derived vaccines have the potential for cost-effective manufacture and are not known to replicate in mammalian cells, they represent a viable alternative to current replicating vaccine vectors for development of both human and veterinary vaccines.

Amino Acid Sequence↗

Cyclospora cayetanensis in three populations at risk in Guatemala.

In 1996 and 1997, outbreaks of Cyclospora cayetanensis in North America were linked to Guatemalan raspberries. From April 1999 to April 2000, we undertook a survey for C. cayetanensis in raspberry farm workers, malnourished children, and human immunodeficiency virus and AIDS patients in Guatemala. Stool samples were analyzed using ethylacetate-formalin concentration, wet preparation, modified acid-fast staining method, and epifluorescence. Oocysts were found in 1.5% of the subjects, none of whom were raspberry farm workers.

AIDS-Related Opportunistic Infections↗

Effect of different baculovirus inactivation procedures on the integrity and immunogenicity of porcine parvovirus-like particles.

We have demonstrated earlier the usefulness of recombinant porcine parvovirus (PPV) virus-like particles (VLPs) as an efficient recombinant vaccine for PPV. Here, we have demonstrated that preparations of PPV VLPs could be contaminated by recombinant baculoviruses. Since these baculoviruses can be a problem for the registration and safety requirements of the recombinant vaccine, we have tested different baculovirus inactivation strategies, studying simultaneously the integrity and immunogenicity of the VLPs. These methods were pasteurization, treatment with detergents and alkylation with binary ethylenimine (BEI). The structural and functional integrity of the PPV VLPs after the inactivation treatments were analyzed by electron microscopy, hemagglutination, double antibody sandwich (DAS)-ELISA and immunogenicity studies. Binary ethylenimine and Triton X-100 inactivated particles maintained all the original structural and antigenic properties. In addition, PPV VLPs were subjected to size-exclusion chromatography to analyze the presence of VP2 monomers or any other contaminant. The resulting highly purified material was used as the standard of reference to quantify PPV VLPs in order to determine the dose of vaccine by DAS-ELISA. After immunization experiments in guinea pigs, the antibody titers obtained with all the inactivation procedures were very similar. Triton X-100 treatment was selected for further testing in animals because of the speed, simplicity and safety of the overall procedure.

Animals↗

Oxidative stress and lipid abnormalities in renal transplant recipients with or without chronic rejection.

BACKGROUND: The histological picture of chronic rejection with endothelial lesions and vascular hyperplasia resembles the early arteriosclerotic lesions. As increasing evidence suggests a role for oxidative stress in arteriosclerosis, we examined whether chronic rejection in renal transplant recipients was associated with increased oxidative stress markers. METHODS: We investigated lipid metabolism and oxidative stress in 77 renal transplant recipients. Group I patients (n=34; 48+/-2 years old, 12 women, 22 men) had no clinical or histological signs of chronic rejection, whereas group II patients (n=43; 47+/-3 years old, 15 women, 28 men) had histologically proven chronic rejection. All patients were treated with cyclosporine and steroids. Lipid metabolism was evaluated by determining total cholesterol, triglycerides, high-density lipoprotein cholesterol, apolipoproteins AI and B, and lipoprotein (a). Oxidative stress was evaluated by determining: (i) the end product of lipid peroxidation, malonyldialdehyde (MDA), and erythrocyte polyunsaturated fatty acids; (ii) the nonenzymatic antioxidant system: erythrocyte alpha-tocopherol and glutathione; and (iii) the enzymatic antioxidant system: erythrocyte superoxide dismutase and plasma glutathione peroxidase. Results were compared with those of a control group (38 healthy volunteers). RESULTS: Compared with controls, renal transplant recipients had significantly increased total cholesterol, triglyceride, and apolipoprotein B levels; they also had, in association with these lipid abnormalities, a significant increase in MDA and a significant decrease in erythrocyte polyunsaturated fatty acids, as well as a significant decrease in enzymatic and nonenzymatic antioxidant defense mechanisms. In contrast to lipid disturbances, where no difference was observed between groups I and II, markers of oxidative stress were significantly higher in group II compared with group I (MDA: 1.87+/-0.43 and 1.62+/-0.31 nmol/ml, respectively, P<0.05). The red blood cell antioxidative defense mechanisms were significantly decreased in group II compared with controls (erythrocyte alpha-tocopherol: 0.61+/-0.38 and 1.08+/-0.31 mg/L, respectively, P<0.01; superoxide dismutase: 1.08+/-0.2 and 1.32+/-0.31 U/mg Hb, respectively, P<0.01). CONCLUSION: Our data show that oxidative stress with a decrease in antioxidant defenses is associated with kidney transplantation. In addition, oxidative stress markers are particularly increased in transplant recipients with chronic rejection, which suggests that oxidative stress may participate in the development and/or progression of vascular lesions observed in these patients.

Adult↗

Long-term improvement in renal function after cyclosporine reduction in renal transplant recipients with histologically proven chronic cyclosporine nephropathy.

BACKGROUND: Chronic cyclosporine (CsA) nephropathy, which has been unequivocally documented in recipients of heart, heart-lung, liver, or bone marrow transplants, as well as in nontransplant situations, usually results in a progressive deterioration of renal function. In this study, we assessed the potential reversibility of chronic CsA nephropathy in renal transplant recipients. PATIENTS AND METHODS: Twenty-three renal transplant patients with biopsy-proven CsA nephropathy associated with long-term CsA administration (27+/-4 months) were followed up for more than 2 years after CsA reduction (18/23 patients) or withdrawal (5/23 patients) and addition of azathioprine. Changes in effective renal plasma flow and glomerular filtration rate were assessed before and 2 years after CsA reduction, whereas serum creatinine, proteinuria, blood pressure, and CsA concentrations were monitored up to 5 years. RESULTS: At 2-year follow-up, glomerular filtration rate increased from 40+/-3 to 47+/-4 (P<0.05) and effective renal plasma flow from 217+/-23 to 244+/-24 ml/min/1.73 m2 (NS). Mean arterial pressure significantly decreased from 98.7+/-2.9 to 93.1+/-2.7 mmHg (P<0.05). There was no significant change in renal vascular resistance, filtration fraction, or albumin excretion. A significant decrease in serum creatinine was also observed during the whole follow-up (73+/-6.5 months). CsA reduction was followed by only one episode of acute reversible rejection; chronic rejection developed in three patients 2 years or later after CsA reduction. CONCLUSIONS: These data suggest that CsA nephropathy participates in graft dysfunction in a small group of renal transplant recipients. In addition, graft dysfunction may be reversible when CsA dosage is reduced early after diagnosis of chronic CsA nephropathy.

Adult↗

Mapping the antigenic structure of porcine parvovirus at the level of peptides.

The antigenic structure of the capsid proteins of porcine parvovirus (PPV) was investigated. A total of nine linear epitopes were identified by Pepscan using porcine or rabbit anti-PPV antisera. No sites were identified with a panel of neutralising monoclonal antibodies (MAbs). All epitopes were located in the region corresponding to the major capsid protein VP2. Based on this information, and on analogy to other autonomous parvoviruses, 24 different peptides were synthesised, coupled to keyhole limpet haemocyanin (KLH) and used to immunise rabbits. Most antisera were able to bind viral protein. Only peptides from the N-terminal part of VP2 were able to induce virus-neutralising antibodies, although at low levels. A similar neutralising activity could be obtained in pigs. The exposure of the N-terminus was shown in full virions, both by immunoelectron microscopy and absorption experiments. It is concluded that in PPV, the VP2 N-terminus is involved in virus neutralisation (VN) and peptides from this region are therefore primary targets for developing peptide-based vaccines against this virus.

Amino Acid Sequence↗

Antigenic structure of the capsid protein of rabbit haemorrhagic disease virus.

Rabbit haemorrhagic disease virus (RHDV) causes an important disease in rabbits. The virus capsid is composed of a single 60 kDa protein. The capsid protein gene was cloned in Escherichia coli using the pET3 system, and the antigenic structure of RHDV VP60 was dissected using 11 monoclonal antibodies (MAbs) and 12 overlapping fragments of the protein expressed in E. coli. Two antigenic regions were found. Ten out of the 11 MAbs recognized different discontinuous epitopes in the most immunodominant region of the viral capsid. This domain was located between residues 31 and 250 of the VP60 N terminus. The other MAb revealed the presence of an antigenic site within 102 aa of the C terminus. This MAb did not recognize the major cleavage product of the full-length 60 kDa protein. These results indicate that, in contrast to other caliciviruses such as Norwalk virus (NV), the 36 kDa cleavage product probably forms the N-terminal region of VP60. However, as in NV, the cleavage region appears to be the most immunodominant region.

Animals↗

Temporary vascular access for extracorporeal renal replacement therapies in acute renal failure patients.

Temporary vascular access is an essential component to perform any extracorporeal renal replacement therapy (RRT) in the acute renal failure patient. RRT used in the acute setting may be categorized in two groups: intermittent (IRRT) and continuous (CRRT). Therapeutic indications are based on clinical and technical considerations. Continuous modalities are mainly utilized in intensive care units for hemodynamically compromised patient. Initially performed spontaneously via an arteriovenous circuit, CRRT modalities have progressively become venovenous with the circulatory assistance of a blood pump. Since both intermittent and continuous RRT modalities are now performed almost exclusively by venovenous modalities, this article deals exclusively with temporary venous catheters. At present, double-lumen catheters represent the most common vascular access for RRT modalities. Semi-rigid polyurethane catheters currently used in case of emergency are limited to short term use. Hemocompatible, flexible silicone catheters, less aggressive for the vessels, seem better suited for the medium and long term run. The tunneled silicone catheters (DualCath type) meet the short and long term needs, and allow for blood flow rates up to 400 ml/min. The internal jugular vein, particularly the right one, seems to warrant the proper functioning of catheters while reducing the risk of stenotic complications. Subclavian access should be limited in time and reserved for silicone catheters in order to limit the risk of stenosis and/or thrombosis. Femoral access, very useful in cases of emergency and respiratory problems, greatly impairs the patient's mobility and should be limited by time to prevent thrombosis and/or infection. Late and/or delayed dysfunctioning of catheters are indicative of a thrombosis. Performance standards of catheters are less of a limiting factor in continuous low flow RRT modalities than in the intermittent ones. Finally, careful handling of the catheter essential to prevent infectious complications.

Acute Kidney Injury↗

[Acute renal insufficiency: nutrition disorders and therapeutic consequences].

Catabolism is usually enhanced in acute renal failure (ARF). Its magnitude varies from one patient to another and can change significantly in the same patient from day to day, reflecting its clinical course. It depends on the severity of the ARF, the underlying process, the associated co-morbidity, and therapeutic approach. The detection of patients at high risk for malnutrition is extremely important; nutritional markers and indexes of caloric and protein requirements are useful to adapt renal replacement and nutritional support to ARF patients. Various biochemical parameters (namely, serum albumin and prealbumin), anthropometic measures, indirect calorimetry, urea and creatinine kinetics are all useful tools to evaluate metabolic status and requirements nutritional. Commonly, the caloric requirements are nearly 35 kcal/kg/24 h with correction factors applied for certain clinical situations: carbohydrates account for 50 to 60% of those needs whereas lipids account for the rest. The total amount of fluid administered has to be adapted to the possible ultrafiltration achieved by dialysis. Daily dialysis sessions and continuous renal replacement therapy allow larger volumes and thus facilitate nutritional support. Protein needs frequently exceed 1.2 g/kg/24 h to maintain the nitrogen balance, with a calorie to protein ration close to 150 kcal per g of nitrogen. Sufficient amounts of vitamins and oligo-elements are necessary. Stimulating anabolism by exogenous mediators, such as androgenic hormones or growth factors (rh-IGF1, rh-GH) is an avenue that deserves better definition in critically ill ARF patients.

Acute Kidney Injury↗

Biochemical and genetic characterization of the acetaldehyde dehydrogenase complex from Acetobacter europaeus.

The aldehyde dehydrogenase complex, which catalyzes the oxidation of acetaldehyde to acetic acid, was purified to apparent homogeneity from the membrane fraction of the industrial vinegar-producing strain Acetobacter europaeus. The determined Km for acetaldehyde was 2.1 mM. SDS-PAGE of the enzyme complex showed the presence of three different subunits with molecular masses of 79, 46, and 17 kDa, respectively. The two larger subunits contained heme. The difference spectrum indicated a cytochrome c, a heme B, and a [2Fe-2S] cluster. The nucleotide sequence of several cloned fragments of a 6-kb chromosomal DNA segment from A. europaeus was determined. It contains three consecutive open reading frames that correspond to proteins with calculated molecular masses of 84.1, 49.0, and 16.7 kDa; these were assigned to the purified proteins and named aldH, aldF, and aldG, respectively. The N-terminal sequence of the 79-kDa subunit was detected within the predicted amino acid sequence of AldH, which indicated the presence of a leader peptide. Cotranscription of the three genes was shown by Northern hybridization. Sequence analysis and experimental evidence allowed the assignment of the following cofactors to the respective subunits of the aldehyde dehydrogenase complex: heme C to AldF, [2Fe-2S] cluster to AldG, and heme B and a molybdopterin cofactor to AldH. Part of an open reading frame, gdhA, was detected upstream of the operon that showed high similarities to the C-terminal part of several pyrroloquinoline-chinone-dependent glucose dehydrogenases.

Acetobacter↗

Baculovirus expression of proteins of porcine reproductive and respiratory syndrome virus strain Olot/91. Involvement of ORF3 and ORF5 proteins in protection.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a new arterivirus that has spread rapidly all around the world in the last few years. The genomic region containing open reading frames (ORFs) 2 to 7 of PRRSV Spanish isolate Olot/91 was cloned and sequenced. The genomic sequence shared 95% identity with Lelystad and Tübingen isolates and between 61-64% with the ORF7 region of the American isolates. ORFs 2 to 7 were inserted into recombinant baculoviruses downstream of the polyhedrin promoter. Only ORFs 2, 3 5 and 7 were expressed in insect cells as detected by PRRS-specific pig antisera. To analyze the immunogenicity of these proteins and their ability to confer protection, Sf9 cells infected with recombinant baculoviruses expressing ORFs 3, 5 and 7 gene products were used to immunize pregnant sows, either individually or in combination. The results obtained indicate that ORFs 3 and 5 gene products could be major candidates for the development of a vaccine against PRRS since they conferred 68.4 and 50% protection, respectively, as evaluated by the number of piglets born alive and healthy at the time of weaning. In addition, piglets born to sows immunized with ORFs 3 and 5 proteins were seronegative to PRRSV after weaning, indicating absence of viral replication. ORF7 is the most immunogenic protein of PRRSV, but the antibodies induced in sows are non-protective and may even interfere with protection.

Animals↗