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Biomedical subjects

M Alvarez

Publications and source records attributed to M Alvarez.

At least 37 records · Page 2Linked to original sources
PubMed2004

Dengue virus type 2 in Cuba, 1997: conservation of E gene sequence in isolates obtained at different times during the epidemic.

It was recently reported that disease severity increased during the 1997 Cuban dengue 2 virus epidemic and it was suggested that this might be explained by the appearance of neutralization resistant escape mutants. We investigated these observations and ideas by sequencing 20 dengue 2 virus isolates obtained during the early (low case fatality rate) and the late (high case fatality rate) phases of the outbreak. Our results showed total conservation of the E gene sequence for these isolates suggesting that the selection of envelope gene escape mutants was not the determinant of increased disease severity. Alignment of these sequences with those available in GenBank, followed by Maximum likelihood phylogenetic analysis generated a tree, which indicated that our isolates are closely related to the virus that circulated in Venezuela in 1997/98 and subsequently in Martinique in 1998. This "American/Asian" genotype has therefore gradually dispersed across the Caribbean region during the past 5 years.

Cuba↗
PubMed2004

Nanomechanics of the formation of DNA self-assembled monolayers and hybridization on microcantilevers.

Biomolecular interactions over the surface of a microcantilever can produce its bending motion via changes of the surface stress, which is referred to nanomechanical response. Here, we have studied the interaction forces responsible for the bending motion during the formation of a self-assembled monolayer of thiolated 27-mer single-stranded DNA on the gold-coated side of a microcantilever and during the subsequent hybridization with the complementary nucleic acid. The immobilization of the single-stranded DNA probe gives a mean surface stress of 25 mN/m and a mean bending of 23 nm for microcantilevers with a length and thickness of about 200 microm and 0.8 microm, respectively. The hybridization with the complementary sequence could not be inferred from the nanomechanical response. The nanomechanical response was compared with data from well-established techniques such as surface plasmon resonance and radiolabeling, to determine the surface coverage and study the intermolecular forces between neighboring DNA molecules anchored to the microcantilever surface. From both techniques, an immobilization surface density of 3 x 10(12) molecules/cm(2) and a hybridization efficiency of 40% were determined. More importantly, label-free hybridization was clearly detected in the same conditions with a conventional sensor based on surface plasmon resonance. The results imply that the nanomechanical signal during the immobilization process arises mainly from the covalent attachment to the gold surface, and the interchain interactions between neighboring DNA molecules are weak, producing an undetectable surface stress. We conclude that detection of nucleic acid hybridization with nanomechanical sensors requires reference cantilevers to remove nonspecific signals, more sensitive microcantilever geometries, and immobilization chemistries specially addressed to enhance the surface stress variations.

Base Sequence↗
PubMed2004

Effect of the interval between estrus onset and artificial insemination on sex ratio and fertility in cattle: a field study.

We have carried out a field trial in cattle to study the effect of the interval between the onset of estrus and AI on sex ratio and fertility. Data were obtained from 716 cows that had been inseminated at different times between 8 and 44 h from the visual detection of estrus. Before analyzing the data, it was grouped in three intervals considering the time between estrus onset and AI (8-18, 18-30, and > or = 30 h). Our results show that the percentage of calved females (73.05%) is significantly superior for early inseminations (8-18 h), and it decreases 1.85% per hour from the onset of estrus. Delayed AIs (> or = 30 h) produce a significant deviation of the sex ratio towards the males (72.06%); nevertheless, fertility (percentage of successful pregnancies) diminishes significantly, from 66.19% (8-18 h) to 45.35% (> or = 30 h). In conclusion, variations in the interval between the onset of estrus and AI modify sex ratio. However, we must consider its effect on fertility.

Animals↗
PubMed2004

Fluid shear stress induces beta-catenin signaling in osteoblasts.

beta-Catenin plays a dual role in cells: one at cell-cell junctions and one regulating gene transcription together with TCF (T-cell Factor) in the nucleus. Recently, a role for beta-catenin in osteoblast differentiation and gene expression has begun to be elucidated. Herein we investigated the effects of fluid shear stress (FSS) on beta-catenin signaling. FSS is a well-characterized anabolic stimulus for osteoblasts; however, the molecular mechanisms for the effects of this stimulation remain largely unknown. We found that 1 hour of laminar FSS (10 dynes/cm(2)) induced translocation of beta-catenin to the nucleus and activated a TCF-reporter gene. Analysis of upstream signals that may regulate beta-catenin signaling activity revealed two potential mechanisms for increased beta-catenin signaling. First, FSS induced a transient, but significant, increase in the phosphorylation of both glycogen synthase kinase 3beta (GSK-3beta) and Akt. Second, FSS reduced the levels of beta-catenin associated with N-cadherin, suggesting that less sequestration of beta-catenin by cadherins occurs in osteoblasts subjected to FSS. Functional analysts of potential genes regulated by beta-catenin signaling in osteoblasts revealed two novel observations. First, endogenous, nuclear beta-catenin purified from osteoblasts formed a complex with a TCF -binding element in the cyclooxygenase-2 promoter, and, second, overexpression of either a constitutively active beta-catenin molecule or inhibition of GSK-3beta activity increased basal cyclooxygenase-2 levels. Together, these data demonstrate for the first time that FSS modulates the activity of both GSK-3beta and beta-catenin and that these signaling molecules regulate cyclooxygenase-2 expression in osteoblasts.

3T3 Cells↗
PubMed2004

Retrospective analysis of antiretroviral HIV treatment success based on medical history or guided by the reverse hybridisation LiPA HIV genotyping system.

The changes in viral load and CD4(+) count at 3 and 6 months in a group of 166 HIV-infected patients was evaluated. The new therapy was chosen based on the medical history procedures for 70 patients, and in 96 patients it was guided by the partial or complete result of the line probe assay (LiPA) HIV RT and Protease resistance tests. The absolute difference from the baseline of the log viral load at 3 and 6 months was significantly different between the two groups when adjusted for baseline viral load (P < 0.0001) and stayed significant when intention-to-treat analysis was carried out (P < 0.001). The absolute difference of the CD4(+) count was not significantly different when adjusted for baseline CD4(+) (P = 0.854, 3 months; P = 0.06, 6 months). The proportion of patients with a viral load </=200 cp/ml in the medical history group (14.5%, 3 months; 15.2%, 6 months) was significantly different from the proportion of responders in the LiPA group (28.7%, 3 months, P = 0.03; 34.7%, 6 months, P = 0.008). In the intention-to-treat population, the difference between the two groups remained significant (P = 0.01). There was no statistical difference between the two groups in terms of adherence (P = 0.88), number of drug failures (P = 0.12) and for the time since starting treatment (in years) (P = 0.48), but there was a significant difference for the number of new drugs in the new regimen (P < 0.0001) and for the pill burden of the treatment (P = 0.0006). A higher antiretroviral HIV treatment success guided by the LiPA HIV genotyping system than that based on the medical history only is reported. Of note, this study reached the same conclusions as previous studies, which all used sequencing.

Adult↗
PubMed2004

[Management of occupational and sexual exposure to HIV in the Toulouse teaching hospital].

OBJECTIVE: This study had aim to describe the management of occupational and sexual HIV exposure in the Toulouse teaching hospital. DESIGN: A prospective descriptive study was made of patients reporting with potential HIV exposure in Toulouse between 01/01/2000 and 12/31/2002. RESULTS: Six hundred and ninety three cases were reported, 236 after occupational and, 457 after sexual exposure. The frequency of sexual exposures increased with time. 61.2% of patients received post-exposure treatment and no seroconversion was diagnosed during their follow-up. Eighty-four percent of treated patients received three anti-retroviral drugs, with a protease inhibitor in 57%. Treatment was more frequently prescribed in sexual exposures than in occupational ones. For occupational exposures, the median time between exposure and consultation was 4 h and was decreased by spontaneous bleeding but not affected by source patient serostatus or injury deepness. Treatment was more frequent when injury was deep, when there was spontaneously bleeding, and when the source patient serostatus was positive or unknown. For sexual exposures, the median time between exposure and consultation was significantly superior to 4 h. That was diminished by positive source person serostatus but not affected by the partner's gender, nature of intercourse, or rape. Treatment was more frequently prescribed in case of positive or unknown source person serostatus, rape and homosexual intercourse. CONCLUSIONS: Given the delay before consultation for sexual exposures and out of delay treatment in occupational exposures, discussion with health professionals on implementing procedures and means seems mandatory.

Adolescent↗
PubMed2004

Transportation of reclaimed wastewater through a long pipe: inhibition of sulphide production by nitrite from the secondary treatment.

The agricultural reuse of reclaimed wastewater has become a necessity in places with water shortages. Frequently, this involves the operation of long transportation pipelines, like in the South Tenerife reuse system, whose main element is a completely filled 61 km long gravity pipe in cast iron. Sulphide generation, which could contribute to pipe corrosion, is a usual process taking place during transportation if anaerobic conditions prevail. In the Wastewater Treatment Plant of Santa Cruz (Tenerife, Spain) a partial nitrification process was achieved by increasing the mean residence time through the aeration step at low dissolved oxygen concentration. Such conditions, combined with the right temperature and a free ammonia concentration above 1 mg l(-1), inhibited nitratation and favoured nitritation, which led to concentrations of NO2-N above 8 mg l(-1) in the secondary effluent. During the transportation, nitrite inhibited the appearance of anaerobic conditions, and, consequently, no sulphide generation occurred. At the same time, a nitrite reduction process took place with a first order kinetics and a rate coefficient of 0.052 h(-1)' at 25 degrees C. A parallel behaviour between the nitrite depletion and the oxidation-reduction potential evolution along the pipeline was also observed.

Ammonia↗
PubMed2004

In vitro activity of moxifloxacin compared to other antimicrobials against streptococci isolated from iatrogenic oral bacteremia in Spain.

BACKGROUND/AIMS: Systemic dissemination of oral bacteria to distant body sites may be the cause of focal infections. The unsuitable use and overexposure to antimicrobial therapy in clinical dental practice may contribute to the worldwide problem of antimicrobial resistance. The aim of this study was to determine the susceptibilities of streptococci isolated from the bloodstream after dental extractions against penicillin, ampicillin, amoxicillin, erythromycin, clindamycin, and a new fluoroquinolone, moxifloxacin. PATIENTS AND METHODS: Eighty-four patients who required dental extractions were studied. Venous blood samples were collected from each patient at baseline (before dental manipulation) and 30 s after dental extractions. The samples were processed in the Bactec 9240. The isolated bacteria were identified by conventional microbiological techniques. The antimicrobial susceptibility of 81 streptococci was determined by the E-test method. The NCCLS performance standards were followed. RESULTS: 88.9-92.5% of the streptococci were sensitive to beta-lactam agents tested with a minimum inhibitory concentration (MIC)(90s) ranging from 0.094 to 0.19 mg/l. The resistance to erythromycin and clindamycin was 40.8% (MIC(90HR) = 256 mg/l) and 21% (MIC(90HR) = 256 mg/l), respectively. The MIC(90) to moxifloxacin was 0.125 mg/l. CONCLUSION: Most of the streptococci isolated from the bloodstream after dental extractions were susceptible in vitro to penicillin, ampicillin, and amoxicillin. The high percentage of streptococci resistant to erythromycin and clindamycin could restrict their usefulness as prophylactic drugs. All the isolates showed a low MIC of moxifloxacin in vitro, making it a promising antimicrobial alternative for the prevention of streptococcal focal infections associated with certain dental manipulations, when the administration of beta-lactam agents is not indicated.

Adolescent↗
PubMed2004

Epidemiology of conjugative plasmid-mediated AmpC beta-lactamases in the United States.

A sample of 752 resistant Klebsiella pneumoniae, Klebsiella oxytoca, and Escherichia coli strains from 70 sites in 25 U.S. states and the District of Columbia was examined for transmissibility of resistance to ceftazidime and the nature of the plasmid-mediated beta-lactamase involved. Fifty-nine percent of the K. pneumoniae, 24% of the K. oxytoca, and 44% of the E. coli isolates transferred resistance to ceftazidime. Plasmids encoding AmpC-type beta-lactamase were found in 8.5% of the K. pneumoniae samples, 6.9% of the K. oxytoca samples, and 4% of the E. coli samples, at 20 of the 70 sites and in 10 of the 25 states. ACT-1 beta-lactamase was found at eight sites, four of which were near New York City, where the ACT-1 enzyme was first discovered; ACT-1 beta-lactamase was also found in Massachusetts, Pennsylvania, and Virginia. FOX-5 beta-lactamase was also found at eight sites, mainly in southeastern states but also in New York. Two E. coli strains produced CMY-2, and one K. pneumoniae strain produced DHA-1 beta-lactamase. Pulsed-field gel electrophoresis and plasmid analysis suggested that AmpC-mediated resistance spread both by strain and plasmid dissemination. All AmpC beta-lactamase-containing isolates were resistant to cefoxitin, but so were 11% of strains containing transmissible SHV- and TEM-type extended-spectrum beta-lactamases. A beta-lactamase inhibitor test was helpful in distinguishing the two types of resistance but was not definitive since 24% of clinical isolates producing AmpC beta-lactamase had a positive response to clavulanic acid. Coexistence of AmpC and extended-spectrum beta-lactamases was the main reason for these discrepancies. Plasmid-mediated AmpC-type enzymes are thus responsible for an appreciable fraction of resistance in clinical isolates of Klebsiella spp. and E. coli, are disseminated around the United States, and are not so easily distinguished from other enzymes that mediate resistance to oxyimino-beta-lactams.

Bacterial Proteins↗
PubMed2003

Differential effect of fetal, neonatal and treatment variables on neurodevelopment in infants with congenital hypothyroidism.

OBJECTIVES: To identify the influence of fetal, neonatal and treatment variables on neurodevelopment in children with congenital hypothyroidism. METHODS: A multiple regression analysis was performed in the neurodevelopmental scores of 26 children with CH who were assessed at 3, 6, 12 and 18 months for postural control, fine eye-hand coordination and language. The independent variables were: duration of fetal hypothyroidism, initial biochemical severity, days at starting treatment, initial dose, and efficiency of treatment. RESULTS: Postural control was predicted by duration of fetal hypothyroidism and language by the biochemical severity of the disease at birth. Fine eye-hand coordination was not predicted by any variable. CONCLUSION: This pattern of influences suggests that fetal and neonatal hypothyroidism have different effects on brain maturation sites during the first 2 years of life and that some subtle neurocognitive deficits are probably unavoidable.

Child Development↗
PubMed2003

Genetic heterogeneity of bovine viral diarrhoea viruses from Spain.

Forty-four bovine viral diarrhoea virus (BVDV) isolates collected from the north of Spain between 1989 and 2000 were characterised at the molecular level. For 24 of these isolates the entire N(pro) gene sequence was determined. Phylogenetic analysis revealed that 21 isolates belong to subgroup BVDV-1b, while two BVDV-1c isolates and one BVDV-1h isolate were found. Furthermore, 20 additional virus isolates were analysed by differential reverse transcription-polymerase chain reaction (RT-PCR) and classified as BVDV-1. The results from our study demonstrate that BVDV-1b is the most prevalent subgroup present in bovine dairy herds of Spain, while there is no evidence for the presence of BVDV-2.

Amino Acid Sequence↗
PubMed2003

Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem.

Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reserves from genetically valuable animals or endangered species. However, there are many factors that influence the outcome of this technique. We have studied the effect of the interval between animal's death and sperm recovery (0, 24 or 48 h) on the quality and freezability of ram spermatozoa from cauda epididymidis. Storage temperature of epididymis (room temperature or 5 degrees C) was also analysed. Spermatozoa were diluted with Tes-Tris-Fructose solution supplemented with egg yolk (10%) and glycerol (4%), and frozen using a programmable biofreezer (-20 degrees C/min). Pre-freeze and post-thaw sperm samples showed viable spermatozoa up to 48 h after the animal's death, although their quality declined significantly as post-mortem storage time increased. Epididymis sperm stored at 5 degrees C showed better motility and a lower percentage of abnormal forms than epididymis stored at room temperature after 24 and 48 h. The fertilizing ability of cauda epididymis ram spermatozoa obtained at 0 and 24h after the animal's death is similar to that of ejaculated spermatozoa. Therefore, a good protocol for post-mortem semen collection in rams when epididymal spermatozoa cannot be collected immediately, is to preserve the epididymis at 5 degrees C and process the samples in the first 24h after the animal's death.

Acrosome↗
PubMed2003

Field and in vitro assay of three methods for freezing ram semen.

Glycerol has been the most widely used cryopreservation agent for spermatozoa and a wide range of factors affect its action on sperm viability and fertilizing capacity. We tested three methods for freezing ram semen packed in 0.25 ml straws (final cellular concentration: 100 x 10(6) spz/ml). Method M1: Two-thirds of the final volume of diluent was added as solution A (without glycerol) to the pure semen at 35 degrees C. The sample was cooled to 5 degrees C (-0.30 degrees C/min), one-third of final diluent volume was added as solution B (final concentration of glycerol 4%) and the sample was maintained at 5 degrees C for 2h. It was then frozen in a programmable biofreezer (-20 degrees C/min down to -100 degrees C). Method M2: The sample was diluted with a specific solution at 35 degrees C (final concentration of glycerol 3%), cooled to 5 degrees C (-0.20 degrees C/min) and left for 2h. After that, it was frozen in nitrogen vapours. Method M3: Semen was diluted 1:1 in a specific solution (concentration of glycerol 2%) and cooled to 5 degrees C (-0.25 degrees C/min). The sample was then diluted again in the same solution to the final cellular concentration (final concentration of glycerol 4%). It was left for 1h at 5 degrees C and then frozen in a programmable biofreezer (-20 degrees C/min down to -100 degrees C). Best total motility (TM) and progressive motility (PM) (75.8 and 55.18%) were obtained using Method M3. Methods M1 and M3 gave significantly higher values (P<0.05) for kinetic parameters: average path velocity (VAP) (81.3 and 85.2 microm/s), straight-line velocity (VSL) (72.8 and 77.3 microm/s) and linearity (LIN) (66.6 and 68.8%). Method M2 showed the lowest kinetic parameters of motility (VAP 74.4, VSL 67.3 and LIN 62.5) and the highest percentage of cells with damaged plasma membrane (53.8%). Method M1 gave the worst results in viability and acrosome status assessed using fluorescence probes (31.3%-dead cells with damaged acrosomes-versus 25.4% in M2 and 23.3% in M3). A field trial carried out on fertility showed a significantly higher percentage of pregnant or lambing ewes (P<0.05) with Method M3 (67.3% versus 51.1% for M1 and 58.8% for M2). We concluded that the use of a simple dilution medium (test-fructose-glycerol-egg yolk) with the addition of glycerol (to 2% at 35 degrees C and to 4% at 5 degrees C) in two steps together with a programmable biofreezer was a productive method for freezing ram semen.

Acrosome↗
PubMed2003

Diagnostic imaging in sheep hepatic fascioliasis: ultrasound, computer tomography and magnetic resonance findings.

Radiological features and biochemical changes were investigated during the parenchymal and ductal phases of chronic Fasciola hepatica infection in sheep. The activities of plasma aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and gamma-glutamyl transpeptidase (GGT), plasma levels of IgG anti- F. hepatica and serial ultrasound, computed tomography (CT) and magnetic resonance imaging (MRI) findings were studied in eight sheep infected with 150 F. hepatica metacercariae. Experimental fluke infection provoked an increase in plasma level of IgG directed against F. hepatica and in plasma LDH and AST activities from 4 weeks after infection. Enzyme activities did not significantly differ from the baseline after 15 and 12 weeks for LDH and AST, respectively. GGT activity increased from 9 weeks postinfection and still remained significantly elevated at 18 weeks. In the parenchymal phase, both CT and MRI showed nodular lesions in five animals and MRI could also detect early tracks in the subcapsular area in three sheep. Ultrasound findings were nonspecific in this phase. Ductal dilatation was shown by ultrasound, CT and MRI in almost all animals, although MRI was inferior to CT in depicting a mild ductal dilatation. Moving echogenic forms in the dilated bile ducts were observed by ultrasound from 9 weeks postinfection in seven of the eight sheep. Moving worms were only demonstrated in four sheep at CT and in a single sheep at MRI. This study shows that radiological findings may be useful in studies of fluke-induced liver damage.

Animals↗
PubMed2003

An increased expression of nucleolin is associated with a physiological nucleolar segregation.

Nucleolar segregation is the most striking cellular phenotypic feature of cold-acclimatized carp and depicts the cyclical reprogramming that the physiology of the fish undergoes between summer and winter, where a clear differential expression of some nucleolar related genes occurs. We characterized carp nucleolin, a nucleolar protein involved in multiple steps of ribosome biogenesis, and evaluated its expression upon fish acclimatization. We show that the carp cDNA deduced amino acid sequence exhibits the same tripartite structural organization found in other species. Nevertheless, we observed that nucleolin mRNA expression was strongly induced in the cold-adapted carp as was the nuclear protein content, assessed by immunocytochemistry in liver sections. The physiological up-regulation of nucleolin in the cold-acclimatized carp, where rRNA transcription and processing are depressed concomitantly with the nucleolus segregation, is consistent with the notion that nucleolin plays a fundamental role in repressing rRNA synthesis.

Acclimatization↗