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Biomedical subjects

C Vaquero

Publications and source records attributed to C Vaquero.

At least 37 records · Page 2Linked to original sources
PubMed1999

Venographic comparison of subcutaneous low-molecular weight heparin with oral anticoagulant therapy in the long-term treatment of deep venous thrombosis.

PURPOSE: The primary objective of this study was to evaluate with venography the rate of thrombus regression after a fixed dose of low-molecular weight heparin (LMWH) per day for 3 months compared with oral anticoagulant therapy for deep venous thrombosis (DVT). Secondary endpoints were the comparisons of the efficacy and safety of both treatments. METHODS: This study was designed as an open randomized clinical study in a university hospital setting. Of the 165 patients finally enrolled in the study, 85 were assigned LMWH therapy and 80 were assigned oral anticoagulant therapy. In the group randomized to oral anticoagulant therapy, the patients first underwent treatment in the hospital with standard unfractionated heparin and then coumarin for 3 months. Doses were adjusted with laboratory monitoring to maintain the international normalized ratio between 2.0 and 3.0. Patients in the LMWH group were administered subcutaneous injections of fixed doses of 40 mg enoxaparin (4000 anti-Xa units) every 12 hours for 7 days, and after discharge from the hospital, they were administered 40 mg enoxaparin once daily at fixed doses for 3 months without a laboratory control assay. A quantitative venographic score (Marder score) was used to assess the extent of the venous thrombosis, with 0 points indicating no DVT and 40 points indicating total occlusion of all deep veins. The rate of thrombus reduction was defined as the difference in quantitative venographic scores after termination of LMWH or coumarin therapy as compared with the scores obtained on the initial venographic results. The efficacy was defined as the ability to prevent symptomatic extension or recurrence of venous thromboembolism (documented with venograms or serial lung scans). The safety was defined as the occurrence of hemorrhages. RESULTS: After 3 months of treatment, the mean Marder score was significantly decreased in both groups in comparison with the baseline score, although the effect of therapy was significantly better after LMWH therapy (49.4% reduction) than after coumarin therapy (24.5% reduction; P <.001). LMWH therapy and male gender were independently associated with an enhanced resolution of the thrombus. A lower frequency of symptomatic recurrent venous thromboembolism was also shown in patients who underwent treatment with LMWH therapy (9.5%) than with oral anticoagulant therapy (23.7%; P <.05), although this difference was entirely a result of recurrence of DVT. Bleeding complications were significantly fewer in the LMWH group than in the coumarin group (1. 1% vs 10%; P <.05). This difference was caused by minor hemorrhages. Coumarin therapy and cancer were independently associated with an enhanced risk of complications. Subcutaneous heparin therapy was well tolerated by all patients. CONCLUSION: The patients who were allocated to undergo enoxaparin therapy had a significantly greater improvement in their quantitative venographic score, a significantly lower recurrence rate of symptomatic venous thromboembolism, and a significantly lower incidence of bleeding than patients who underwent treatment with coumarin. LMWH can be used on an outpatient basis as a safer and more effective alternative to classical oral anticoagulant therapy for the secondary prophylaxis of selected patients with DVT.

Adult↗
PubMed1999

Rice cell culture as an alternative production system for functional diagnostic and therapeutic antibodies.

We investigated the suitability of transformed rice cell lines as a system for the production of therapeutic recombinant antibodies. Expression constructs encoding a single-chain Fv fragment (scFvT84.66, specific for CEA, the carcinoembryonic antigen present on many human tumours) were introduced into rice tissue by particle bombardment. We compared antibody production levels when antibodies were either secreted to the apoplast or retained in the endoplasmic reticulum (ER) using a KDEL retention signal. Production levels were up to 14 times higher when antibodies were retained in the ER. Additionally, we compared construct sencoding different leader peptides (plant codon optimised murine immunoglobulin heavy and light chain leader peptides from mAb24) and carrying alternative 5' untranslated regions (the petunia chalcone synthase gene 5' UTR and the tobacco mosaic virus omega sequence). We observed no significant differences in antibody production levels among cell lines transformed with these constructs. The highest level of antibody production we measured was 3.8 micrograms g-1 callus (fresh weight). Immunological analysis of transgenic rice callus confirmed the presence of functional scFvT84.66. We discuss the potential merits of cell culture for the production of recombinant antibodies and other valuable macromolecules.

5' Untranslated Regions↗
PubMed1999

Superficial femoral artery as a substitute for common carotid artery affected by malignant cervical tumors.

Tumoral disease of the arteries is uncommon. It is possible that, in tumoral reoperations affecting cervical structures, periarterial fibrosis due to radiotherapy and previous surgery and metastatic adherences to the arterial wall makes radical surgery difficult. We present a case where the surgical treatment of a regional relapse from laryngeal neoplasm needed the removal of the common carotid artery and its reconstruction with autologous patent superficial femoral artery. Surgical technique included a PTFE graft interposition in the leg.

Carcinoma, Squamous Cell↗
PubMed1998

Do natural antisense transcripts make sense in eukaryotes?

The existence of naturally occurring antisense RNAs has been illustrated, in eukaryotes, by an increasing number of reports. The following review presents the major findings in this field, with a special focus on the regulation of gene expression exerted by endogenous complementary transcripts. A large variety of eukaryotic organisms, contains antisense transcripts. Moreover, the great diversity of genetic loci encoding overlapping sense and antisense RNAs suggests that such transcripts may be involved in numerous biological functions, such as control of development, adaptative response. viral infection. The regulation of gene expression by endogenous antisense RNAs seems of general importance in eukaryotes as already established in prokaryotes: it is likely to be involved in the control of various biological functions and to play a role in the development of pathological situations. Several experimental evidences for coupled, balanced or unbalanced expression of sense and antisense RNAs suggest that antisense transcripts may govern the expression of their sense counterparts. Furthermore, documented examples indicate that this control may be exerted at many levels of gene expression (transcription, maturation, transport, stability and translation). This review also addresses the underlying molecular mechanisms of antisense regulation and presents the current mechanistic hypotheses.

Animals↗
PubMed1998

Monitoring of evoked potentials during spinal cord ischaemia: experimental evaluation in a rabbit model.

OBJECTIVES: Somatosensory evoked potentials (SEPs), spinal evoked potentials (Spinal-EPs), and motor-evoked potentials (MEPs) were monitored in a rabbit model of spinal cord ischaemia to evaluate their accuracy and relationship to clinical status. METHODS: A modified rabbit spinal cord ischaemia model of infrarenal aortic occlusion for 21 min was employed (30 rabbits). After baseline SEPs, Spinal-EPs, and MEPs were obtained, evoked potentials were recorded continuously during and after clamping of the aorta (30 min). Neurological outcome at 24 h was correlated with evoked potentials, and histopathological findings. RESULTS: Fifteen animals became paraplegic. MEPs were always abolished after clamping of the aorta while Spinal-EPs and SEPs remained. The sensory evoked potentials (SEPs and Spinal-EPs) were the least sensitive to spinal cord ischaemia, and their presence had no correlation with the final clinical status (50% of false negatives). This was consistent with histopathological examination that showed damage almost entirely confined to the anterior horn, while the dorsal columns were generally well preserved. High spine MEPs evoked by twitch stimulation was the best predictor of clinical outcome (0% of false negatives, 0% of false positives). CONCLUSIONS: SEPs and Spinal-EPs cannot be used as safe monitors of ischaemia of the spinal cord. High spine MEPs evoked by twitch stimulation was the most useful for real-time evaluation of spinal cord ischaemia, and the best predictor of neurologic outcome during reperfusion.

Animals↗
PubMed1998

[Microsurgical approach and graft anastomosis to the trunk of the facial nerve in the Wistar rat].

An experimental model was developed for the purpose of investigating neural regeneration of the facial nerve in Wistar rats. The anatomic references used to locate the facial nerve at its exit from the skull base are described. A detailed description of the microsurgical anastomosis of cryopreserved nerve allografts is given. The major advantages and disadvantages of rat models compared with other animal models for experimental studies of the facial nerve are discussed.

Anastomosis, Surgical↗
PubMed1997

Analysis of interactions between huGATA-3 transcription factor and three GATA regulatory elements of HIV-1 long terminal repeat, by surface plasmon resonance.

Relative affinities of transcriptional regulatory elements for their respective factor have been essentially studied by bandshift analysis. Here we report a real-time study of factor/DNA interactions using a surface plasmon resonance approach and further characterization of recovered proteins involved in this interaction. For this purpose, human GATA-3, either recombinant or in nuclear extracts, and three natural GATA elements of the HIV-1 long terminal repeat (sites 1, 2, and 3) were chosen, in which only site 2 is a noncanonical GATA site. Direct analysis of sensorgrams, with recombinant huGATA-3, allowed the comparison of association and dissociation profiles of the three DNA regions and their ranking according to their relative affinities. This result, confirmed by competitions with each GATA site, demonstrated the higher relative affinity (at least sevenfold) of site 3. Interactions between the canonical and unique GATA site 3 and nuclear extracts were also studied in real time and provided information on its association and dissociation rates for native huGATA-3. Finally, recovered protein was identified as genuine huGATA-3 by SDS-PAGE, Western blotting, and bandshift assays.

Base Sequence↗
PubMed1997

Regulation of p56(lck) messenger turnover upon T cell activation: involvement of the 3' untranslated region in stability as determined in cell-free extracts.

Full activation of T lymphocytes transiently downregulates the steady state level of the tyrosine kinase p56(lck) mRNA. Here, we show that a decrease in messenger stability is involved in this downmodulation followed thereafter by a rapid and marked increase in mRNA half-life. In order to facilitate the study of p56(lck) messenger stability, an in vitro mRNA decay assay was developed and used to determine whether the 451 nucleotide long 3' untranslated region (3'UTR) of the messenger is implicated in the regulation of mRNA stability. Indeed, deletion of most of the 3'UTR led to a substantial increase in transcript half-life whereas deletion of a limited 3' portion did not, thus showing that the 146 nucleotides located in 5' of the 3'UTR contain destabilizing elements. Furthermore, the stability of both truncated transcripts was still modulated upon activation, thereby suggesting that the activation-responsive elements are located in a region distinct from the 3'UTR.

Cell-Free System↗
PubMed1997

The coat protein is required for the elicitation of the Capsicum L2 gene-mediated resistance against the tobamoviruses.

In Capsicum, the resistance against tobamoviruses conferred by the L2 gene is effective against all but one of the known tobamoviruses. Pepper mild mottle virus (PMMoV) is the only virus which escapes its action. To identify the viral factors affecting induction of the hypersensitive reaction (HR) mediated by the Capsicum spp. L2 resistance gene, we have constructed chimeric viral genomes between paprika mild mottle virus (PaMMV) (a virus able to induce the HR) and PMMoV. A hybrid virus with the PaMMV coat protein gene substituted in the PMMoV-S sequences was able to elicit the HR in Capsicum frutescens (L2L2) plants. These data indicate that the sequences that affect induction of the HR mediated by the L2 resistance gene reside in the coat protein gene. Furthermore, a mutant that codes for a truncated coat protein was able to systemically spread in these plants. Thus, the elicitation of the host response requires the coat protein and not the RNA.

Amino Acid Sequence↗
PubMed1997

Expression of naturally occurring antisense RNA inhibits human immunodeficiency virus type 1 heterologous strain replication.

Recently, the presence of human immunodeficiency virus type 1 (HIV-1) RNA transcripts with negative-strand polarity has been shown in tissue culture models of acute and persistently infected cells. One of these transcripts encodes a 189 amino acid open reading frame. This highly conserved antisense sequence is complementary to the structured Rev-responsive element and extends through the cleavage site of the Env protein. We tested the ability of this antisense RNA to modulate HIV-1 replication and the mRNA profile when expressed stably or transiently in several cell types. Different cell lines and PBLs were transduced by retroviral vectors producing antisense RNA and were then challenged by HIV infection. We have shown that the endogenously expressed antisense RNA containing the natural open reading frame inhibits HIV-1(IIIB) and HIV-1(NDK) replication in these cells. The level of inhibition varied according to the cells, but was significant in all cases. The production of HIV-1 (BRU, IIIB, NDK) mRNAs was also significantly decreased. HIV-2 replication was not inhibited by expression of the antisense RNA. Our results also suggest that this inhibitory effect is due to the antisense RNA and not to the protein which is encoded by this sequence.

Cells, Cultured↗
PubMed1997

Mapping of the RNA-binding domain of the cucumber mosaic virus movement protein.

A series of in-frame deletion mutants was used to identify a domain within the 3a protein of cucumber mosaic virus (CMV) that is required for RNA-binding activity. Deletions in the 3a gene were generated by PCR and restriction digestion, and the resulting mutated 3a sequences were cloned either in pT7-7 or in pGEX-5X3 expression vectors. The mutated 3a proteins or fusions with glutathione S-transferase (GST) were expressed in E. coli, purified, and their nucleic acid-binding activities analysed by photochemical UV cross-linking assays using digoxigenin-UTP-labelled RNA probes. Comparative analyses of seven mutated 3a proteins obtained from inclusion bodies and eight GST fusion proteins revealed that there is an RNA-binding domain located between amino acids 174 and 233. This RNA-binding domain is able to bind single-stranded RNA out of the context of the complete 3a movement protein and is highly conserved within both subgroups of CMV.

Amino Acid Sequence↗
PubMed1996

Accumulation kinetics of CMV RNA 3-encoded proteins and subcellular localization of the 3a protein in infected and transgenic tobacco plants.

The complete nucleotide sequence of RNA 3 of a Spanish isolate of cucumber mosaic virus (CMV-24) has been determined. The encoded putative cell-to-cell movement protein (3a protein) and the coat protein are 279 and 218 amino acids long, respectively. The 3a protein was expressed in Escherichia coli using the vector pT7-7 and was used to raise an immunoserum. We have followed the time course of accumulation of the 3a protein, in parallel to that of the coat protein, and its subcellular localization as a function of time after CMV-24 infection on tobacco plants. The maximum accumulation level of the 3a protein was reached at early stages of infection, being detected in the cytosolic and the cell wall fractions. At later stages of infection, a decline in accumulation levels of the 3a protein was observed, and the protein was essentially associated with the cell wall fractions. These data were corroborated by immunocytochemistry performed in both infected and 3a-expressing transgenic tobacco plants.

Base Sequence↗
PubMed1996

Protective effect of a platelet-activating factor antagonist (WEB-2086) in postischemic renal failure.

The purpose of this study was to evaluate the renoprotective effect of a specific platelet-activating factor antagonist (WEB-2086) in an experimental model of normothermic renal ischemia. Twenty New Zealand white rabbits were studied for 2 days before and 24 hours after a 60-minute period of renal ischemia induced by bilateral clamping of the renal arteries. The animals were divided into two groups: a control group (group A; n = 10) and a treated group (group B; n = 10). In group A the urinary flow rate decreased significantly (from 0.098 +/- 0.008 ml/min to 0.029 +/- 0.005 ml/min) (p < 0.001) and there was a significant reduction in creatinine clearance (from 11.4 +/- 1.2 ml/min to 3.4 +/- 1.1 ml/min) (p < 0.001). In group B no significant changes were observed, although the urinary flow rate increased even in the postischemic period (from 0.09 +/- 0.008 ml/min to 0.11 +/- 0.02 ml/min). Microcirculatory cortical flow showed a postischemic reduction in both groups, although it was most significant in the control group (group A = 43.7%, group B = 71.5%; p < 0.001). Histologic study showed mild damage with patchy tubular necrosis in both groups, although this injury was less severe in the treated group. The results suggest that the preoperative administration of WEB-2086 produces a potent diuretic effect with significant attenuation of postischemic acute renal failure.

Acute Kidney Injury↗
PubMed1996

Epidural regional hypothermia for prevention of paraplegia after aortic occlusion: experimental evaluation in a rabbit model.

PURPOSE: The efficacy of epidural regional hypothermia in the prevention of acute and delayed-onset paraplegia, as well as possible complications and limitations of this technique to a clinically acceptable form, were evaluated in 49 New Zealand white rabbits. METHODS: A modified rabbit spinal cord ischemia model of infrarenal aortic occlusion for 30 minutes was employed. The study was performed in two phases. In phase I (n=20), regional hypothermia induced by epidural perfusion of iced normal saline solution (4 degrees C) was tested versus control in 10 rabbits each (groups A and B). In phase II (n=29) the animals were subdivided into three groups to study the kinetics of absorbtion and distribution of methylene blue (group C; n=10), radiographic contrast material (group D; n=9), and measurement of cerebrospinal pressure while an epidural iced solution was or was not infused (group E; n=10). RESULTS: At 24 and 48 hours, all of the normothermic animals showed irreversible paraplegia (Tarlov score 0). In contrast, at 24 hours none of the rabbits undergoing epidural cold infusion were paraplegic, although at 48 hours one animal had weakness of a hindlimb (Tarlov score 3). Plasma concentration-time profiles of a continuous epidural perfusion with methylene blue showed that the spinal canal is a highly compliant space. Epidurographs showed that epidural perfusion tends to spread more in a cephalic than caudal direction and the main uptake is by the vascular compartment. Despite the large volumes infused (78.75 ml/hr; range, 50 to 100 ml), we observed only a modest transient increase in cerebrospinal fluid pressure (from 2.5 +/- 0.3 mm Hg to 5.4 +/- 0.1 mm Hg), although some animals had intracranial hypertension. CONCLUSIONS: Regional hypothermia induced by epidural cold perfusion has a highly protective effect against the ischemic spinal cord damage. However, this method probably does not avoid the risk of delayed-onset paraplegia. An important limitation of this technique is the difficulty of controlling the intrathecal pressures.

Animals↗
PubMed1996

[Effects of neurotensin on the development of suckling rats with intestinal resection].

Massive intestinal resection produces malabsorption which, in the suckling rat, reduces growth. Our aim was to determine whether the proliferative action of neurotensin, can reduce the negative effects on growth induced by bowel resection. Fifteen days old suckling Wistar rats were used. Twenty rats underwent 90% midgut resection and twelve were used as controls. Half the animals were treated with neurotensin (600 micrograms/kg-day) until sacrifice 30 days later. Body and bone weight were measured and mucosal samples obtained. All resected animals lost body weight and bone weight. Neurotensin treatment reduced femur weight loss. After bowel resection, significant trophic effects were observed at mucosal level (crypt and villous size) but only in the jejunum of resected animals neurotensin treatment had a trophic effect. In conclusion, neurotensin favors intestinal adaptation after resection without improving mid-term growth in the suckling rat.

Adaptation, Physiological↗
PubMed1995

The Capsicum L3 gene-mediated resistance against the tobamoviruses is elicited by the coat protein.

The L3 gene is responsible for the hypersensitive response in Capsicum plants against infection by tobamoviruses. The resistance conferred by this gene is one of the most effective so far described against tobamoviruses. Certain isolates of pepper mild mottle virus (PMMV) are the only tobamoviruses able to overcome the L3 resistance. Chimeric viral genomes between PMMV-S (to which L3 plants are hypersensitive) and PMMV-I (an L3 resistance-breaking isolate) led us to conclude that sequence variation within the coat protein gene of both isolates determines their different virulence in L3L3 plants. Furthermore, the results indicate that a single amino acid substitution, Asn to Met, at position 138 of the PMMV-I coat protein is sufficient to induce the hypersensitive response and localization of viral infection in C. chinense plants. Finally, the use of a mutant coding for a truncated coat protein (maintaining the Met138 coding sequence at the RNA level) demonstrates that a functional coat protein is required for elicitation of the L3 gene-mediated resistance.

Base Sequence↗