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Biomedical subjects

A Alonso

Publications and source records attributed to A Alonso.

At least 19 recordsLinked to original sources

Experimental design to optimise the analysis of organic volatile compounds in cow slurry by headspace solid-phase microextraction-gas chromatography-mass spectrometry.

The headspace solid-phase microextraction (HS-SPME) of some odorous organic compounds [volatile fatty acids (VFAs), phenols and indoles] from animal wastes was optimised using an experimental design approach. The variables affecting the HS-SPME process studied were as follows: type of fibre, pH of the slurry (3-7), addition of sodium chloride (0-1 mol L(-1)), sample volume (10-30 mL), exposure time (10-30 min), exposure temperature (25-50 degrees C), desorption time (3-10 min) and desorption temperature (230-310 degrees C). As for the sorbent, three different types of fibres were studied: 50/30 microm divinylbenzene/Carboxen/polydimethylsiloxane (DVB/CAR/PDMS), 100 microm PDMS and 85 microm CAR/PDMS. Methylation VFAs was observed when fibres containing PDMS were used. The analytes studied in the optimisation procedure were acetic acid, propanoic acid, butanoic acid, 3-methylbutanoic acid, pentanoic acid, phenol, 4-methylphenol, 4-ethylphenol, indole and skatole (3-methylindole). In order to study the influence of the variables on the responses of the compounds, a Plackett-Burman design was built using the Unscrambler program. The optimisation was carried out using real samples. In order to take into account the matrix effect on the calibration curves, standard solutions were prepared using a purged cow slurry free from the analytes of interest. Repeatability within and among days, accuracy and detection limits were calculated from these calibration curves.

Animals↗

Tolerogenic response: allorecognition pathways.

The induction of immune tolerance in transplant recipients has been sought for many years but only a fundamental understanding of the immunological mechanisms underlying graft rejection will allow manipulation of the anti-graft immune response. In general, acute rejection is better understood and treated than chronic rejection, as they occur through partially different mechanisms. It is now generally accepted that recognition of same-species, non-self antigens (allorecognition) occurs through at least two different mechanisms, the direct and indirect pathways. In the direct pathway, donor MHC molecules on the surface of donor antigen-presenting cells (APCs) are recognised directly by the recipient's T cells. This mechanism is so immediate that it seems to be primarily involved in acute graft rejection. Since APCs of donor origin are depleted with time a second mechanism, the indirect pathway, takes over to cause chronic rejection, in which foreign MHC molecules are internalised, partially digested and presented as peptides to recipient T cells. Nonetheless, a number of studies are only fully understood when a third proposed allorecognition mechanism is taken into account. This is the semi-indirect pathway, as discussed in this short report.

Antigen-Presenting Cells↗

Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group.

The mitochondrial DNA (mtDNA) working group of the GEP-ISFG (Spanish and Portuguese Group of the International Society for Forensic Genetics) carried out an inter-laboratory exercise consisting of the analysis of mtDNA sequencing patterns in mixed stains (saliva/semen and blood/semen). Mixtures were prepared with saliva or blood from a female donor and three different semen dilutions (pure, 1:10 and 1:20) in order to simulate forensic casework. All labs extracted the DNA by preferential lysis and amplified and sequenced the first mtDNA hypervariable region (HVS-I). Autosomal and Y-STR markers were also analysed in order to compare nuclear and mitochondrial results from the same DNA extracts. A mixed stain prepared using semen from a vasectomized individual was also analysed. The results were reasonably consistent among labs for the first fractions but not for the second ones, for which some laboratories reported contamination problems. In the first fractions, both the female and male haplotypes were generally detected in those samples prepared with undiluted semen. In contrast, most of the mixtures prepared with diluted semen only yielded the female haplotype, suggesting that the mtDNA copy number per cell is smaller in semen than in saliva or blood. Although the detection level of the male component decreased in accordance with the degree of semen dilution, it was found that the loss of signal was not consistently uniform throughout each electropherogram. Moreover, differences between mixtures prepared from different donors and different body fluids were also observed. We conclude that the particular characteristics of each mixed stain can deeply influence the interpretation of the mtDNA evidence in forensic mixtures (leading in some cases to false exclusions). In this sense, the implementation of preliminary tests with the aim of identifying the fluids involved in the mixture is an essential tool. In addition, in order to prevent incorrect conclusions in the interpretation of electropherograms we strongly recommend: (i) the use of additional sequencing primers to confirm the sequencing results and (ii) interpreting the results to the light of the phylogenetic perspective.

Blood↗

Determination of the DNA-binding characteristics of ethidium bromide, proflavine, and cisplatin by flow injection analysis: usefulness in studies on antitumor drugs.

Flow injection analysis was used to study the reactions occurring between DNA and certain compounds that bind to its double helix, deforming this and even breaking it, such that some of them (e.g., cisplatin) are endowed with antitumoral activity. Use of this technique in the merging zones and stopped-flow modes afforded data on the binding parameters and the kinetic characteristics of the process. The first compound studied was ethidium bromide (EtdBr), used as a fluorescent marker because its fluorescence is enhanced when it binds to DNA. The DNA-EtdBr binding parameters, the apparent intrinsic binding constant (0.31+/-0.02 microM(-1)), and the maximum number of binding sites per nucleotide (0.327+/-0.009) were determined. The modification introduced in these parameters by the presence of proflavine (Prf), a classic competitive inhibitor of the binding of EtdBr to the DNA double helix, was also studied, determining the value of the intrinsic binding constant of Prf (K(Prf) = 0.119+/-9x10(-3) microM(-1)). Finally, we determined the binding parameters between DNA and EtdBr in the presence of the antitumor agent cisplatin, a noncompetitive inhibitor of such binding. This provided information about the binding mechanism as well as the duration and activity of the binding of the compound in its pharmacological use.

Antineoplastic Agents↗

Flow-injection solvent extraction without phase separation. Fluorimetric determination of thiamine by the thiochrome method.

Two modes of liquid-liquid extraction in flow-injection systems were compared and applied to the fluorimetric determination of thiamine (Vitamin B(1)). The first included phase segmentation, but fluorescence was measured without phase separation. In this mode, thiamine was detected at concentrations higher than 8 microg/l with a linear application range of 30-2,000 microg/l, an R.S.D. of 1.9% (150 microg/l, n=10) and a sampling frequency of 60/h. In the second mode, a single segment of organic solution was injected into the aqueous stream and fluorescence was also measured without phase separation. Using this mode, concentrations of thiamine higher than 1 microg/l were detected, with a linear application range between 5 and 280 microg/l, an R.S.D. of 2.4% (150 microg/l, n=10) and a sampling frequency of 60/h. The two forms were applied to the analysis of thiamine in pharmaceuticals.

Flow Injection Analysis↗

Allele frequencies of six miniSTR loci (D10S1248, D14S1434, D22S1045, D4S2364, D2S441 and D1S1677) in a Spanish population.

Allele frequencies and forensic parameters for six miniSTR autosomal loci (D10S1248, D14S1434, D22S1045, D4S2364, D2S441 and D1S1677) were obtained from a sample of 264 unrelated individuals from Spain. No significant deviations from Hardy-Weinberg expectations were found. Due to the small PCR products (<125 bp), the use of these non-CODIS (NC) miniSTRs can increase the probability that a degraded sample can be typed. Additionally, these systems can be used in routine paternity analyses where more markers are needed to increase the power of exclusion or in complex paternity cases (e.g. involving closely related individuals).

DNA Fingerprinting↗

Integrated care prevents hospitalisations for exacerbations in COPD patients.

Hospital admissions due to chronic obstructive pulmonary disease (COPD) exacerbations have a major impact on the disease evolution and costs. The current authors postulated that a simple and well-standardised, low-intensity integrated care intervention can be effective to prevent such hospitalisations. Therefore, 155 exacerbated COPD patients (17% females) were recruited after hospital discharge from centres in Barcelona (Spain) and Leuven (Belgium). They were randomly assigned to either integrated care (IC; n = 65; age mean+/-sd 70+/-9 yrs; forced expiratory volume in one second (FEV(1)) 1.1+/-0.5 L, 43% predicted) or usual care (UC; n = 90; age 72+/-9 yrs; FEV(1) 1.1+/-0.05 L, 41% pred). The IC intervention consisted of an individually tailored care plan upon discharge shared with the primary care team, as well as accessibility to a specialised nurse case manager through a web-based call centre. After 12 months' follow-up, IC showed a lower hospitalisation rate (1.5+/-2.6 versus 2.1+/-3.1) and a higher percentage of patients without re-admissions (49 versus 31%) than UC without differences in mortality (19 versus 16%, respectively). In conclusion, this trial demonstrates that a standardised integrated care intervention, based on shared care arrangements among different levels of the system with support of information technologies, effectively prevents hospitalisations for exacerbations in chronic obstructive pulmonary disease patients.

Aged↗

Evaluation of CD7 and terminal deoxynucleotidyl transferase (TdT) expression in CD34+ myeloblasts from patients with myelodysplastic syndrome.

There is an emerging use of flow cytometry to evaluate patients with myelodysplastic syndrome (MDS). We have studied CD7 and TdT expression in the CD34+ myeloid blast cell population in 55 bone marrow samples of patients with MDS. CD7 and/or TdT were detected in 38 out of 55 patients (69%). CD7 expression was not related to other bad prognosis data but conversely, we found an association between TdT+ CD34 myeloblasts and high-risk MDS patients according to the International Prognostic Scoring System. Therefore, CD7 and TdT may help to establish the diagnosis of MDS and, TdT expression also seems to be a useful marker in distinguishing risk groups.

Adult↗

Differential analysis of donor characteristics for pancreas and islet transplantation.

Suitable selection of donors is key to the success of human islet isolation and transplantation. Although several important donor-related factors have been identified previously, they needed to be confirmed in our setting. The aims of this study were: (1) to compare the characteristics of islet donors with those of pancreas donors (national transplant registry). (2) to compare the characteristics of islet donors resulting in a successful isolation in our facility with the characteristics of pancreas donors, and (3) to compare the characteristics of islet donors at this facility, whether or not isolation was successful, with donors elsewhere whose islets were transplanted and included in the Collaborative Islet Transplant Registry. The 35 islet isolations completed at our facility were analyzed for various characteristics. Significant differences were seen in donor age body mass index (BMI), and body weight between our islet donors and our pancreas donors (P < .001). These differences were maintained in the subgroup analysis corresponding to donors of successful isolations compared to pancreas donors (P < .01). Most successful isolations in our islet isolation facility were associated with donors of BMI >25. The percentage of successful isolation (>300,000 IEq) was higher among donors with a body weight >90 kg. We concluded that there was little overlap between the donor profiles for pancreas transplantation and for islet transplantation. More specific selection criteria relative to both BMI and body weight for islet donors may result in greater success of pancreas islet isolation and transplantation.

Adult↗

Association of increased fibrinogen concentration with impaired activation of anticoagulant protein C.

BACKGROUND: Low levels of activated protein C (APC) are a risk factor for venous thrombosis. The mechanisms leading to interindividual differences in APC are not totally elucidated. Protein C is activated by the thrombin-thrombomodulin complex. As thrombin binds to fibrinogen and thrombomodulin through a common region, it is conceivable that fibrinogen influences the activation of protein C. This would help to explain the association between high levels of fibrinogen and an increased thrombotic risk. METHODS: We analyzed the association between circulating APC levels and fibrinogen concentration in 382 healthy subjects. Subsequently, we studied the effect of increasing fibrinogen concentrations on the APC generation on cultured endothelial cells. RESULTS: An independent inverse association between circulating APC levels and fibrinogen was found [betacoefficient, -0.16; 95% confidence interval (95% CI) -0.26, -0.06; P = 0.001]. For each 100 mg dL(-1) increase in fibrinogen, the independent risk of having low APC levels (<0.7 ng mL(-1)) was almost three times higher (OR 2.8; 95% CI 1.1, 7.2; P = 0.04). Accordingly, a notable association between increasing fibrinogen concentrations and the reduction in the thrombin-thrombomodulin dependent activation of protein C on endothelial cells was found (r = -0.57; P = 0.002). CONCLUSIONS: We present evidence of an inverse association between circulating APC and fibrinogen levels. According to this finding together with the results of our in vitro experiments, we propose that the impairment in the generation of APC on endothelial cells constitutes a new prothrombotic mechanism of fibrinogen.

Adult↗

A comparison of information on motor vehicle crashes as reported by written or telephone interviews.

OBJECTIVE: To compare information about traffic crash injuries and kilometers driven reported in a written questionnaire with information reported in a telephone interview. DESIGN: Telephone and paper surveys. SETTING: The Seguimiento Universidad de Navarra (SUN, University of Navarra Follow-up) study, in Spain. The SUN study is an open enrollment cohort study with 17 000 enrolled graduates followed through biennial mailed questionnaires. SUBJECTS: A sample of 542 individuals from the SUN study participants. MAIN OUTCOME MEASURE: Agreement on information about traffic crash injuries and mileage driven in a mailed questionnaire and a telephone survey. RESULTS: Participation was 90.4%. Considering the phone survey as the gold standard, data on traffic crash injuries in the mailed questionnaire had 83% sensitivity (95% CI 77% to 89%), 77% specificity (95% CI 71% to 82%), 74% positive predictive value (95% CI 67% to 80%), and 89% negative predictive value (95% CI 83% to 93%). Agreement beyond chance, measured by the kappa statistic, was 0.63 (95% CI 0.56 to 0.70). Correlation between questionnaire and telephone surveys and kilometers driven on average during a year assessed by the intraclass correlation coefficient was 0.64 (95% CI 0.57 to 0.70), p<0.001. CONCLUSIONS: Information on sustained traffic crash injuries and traveled mileage over the previous two years as reported through mailed questionnaires in a highly educated population could be used in the study of associations between traffic crash injuries and a variety of risk factors.

Accidents, Traffic↗

[Infection by Epstein-Barr virus and multiple sclerosis].

INTRODUCTION: Multiple sclerosis is the most important disabling neurological disease in young adulthood in our environment. However, its etiology remains unknown. It has been proposed that multiple sclerosis is caused by an interaction of genetic and environmental factors. In particular, Epstein-Barr virus infection could play a fundamental role in the pathophysiology of multiple sclerosis. METHODS: We have reviewed the most recent and important bibliography, both from epidemiologic and basic research studies, dealing with the possible association between Epstein-Barr virus infection and multiple sclerosis. RESULTS: Retrospective and prospective epidemiologic studies suggest a direct association between Epstein-Barr virus infection and the risk of multiple sclerosis. Several physiopathologic mechanisms, including molecular mimicry and bystander activation, could be responsible for this association. CONCLUSIONS: Based on the published evidence, Epstein-Barr virus infection can be considered to have a causal role in the etiology of multiple sclerosis. Further research should be conducted to elucidate the mechanisms underlying this association and to assess its potential for multiple sclerosis prevention and treatment.

Epidemiologic Studies↗

Simultaneous allergy to vine pollen and grape.

We report the case of an 18-year-old female student suffering from seasonal rhinoconjunctivitis with sensitization to pollens from vine and also from grass, olive, and Chenopodiaceae plants who had recently developed episodes of itching, maculopapular rash, and facial angioedema after eating grapes. Testing revealed positive reactions to vine pollen and grapes, and specific IgE were found for both allergens. Immunoblotting and inhibition assays revealed cross-reactivity between the allergenic structures of vine pollen and grape fruit and also among botanically unrelated pollens.

Adolescent↗

Biocompatible hemodialysis membranes for acute renal failure.

BACKGROUND: Acute renal failure (ARF) is associated with substantial morbidity and mortality. Some trials have reported a survival advantage among patients dialyzed with biocompatible membranes (BCM) compared to bioincompatible membranes (BICM). These findings were not consistently observed in subsequent studies. OBJECTIVES: To ascertain whether the use of BCM confers an advantage in either survival or recovery of renal function over the use of BICM in adult patients with ARF requiring intermittent hemodialysis. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL, in The Cochrane Library - Issue 1, 2004), MEDLINE (1966 to January 2004), EMBASE (1980 to January 2004), the Mexican Index of Latin American Biomedical Journals IMBIOMED (1990 to January 2004), the Latin American and Caribbean Health Sciences Literature Database LILACS (1982 to January 2004), and reference lists of articles. SELECTION CRITERIA: Randomized and quasi-randomized controlled trials comparing the use of a BCM with a BICM in patients > 18 years of age with ARF requiring intermittent hemodialysis. DATA COLLECTION AND ANALYSIS: Two authors extracted the data independently. Cellulose-derived dialysis membranes were classified as BICM, and synthetic dialyzers were considered as BCM. The main outcomes were all-cause mortality and recovery of renal function by type of dialyzer. We further explored these outcomes according to the flux properties (high-flux or low-flux) of each of these dialyzers. A meta-analysis was conducted by combining data using a random-effects model. MAIN RESULTS: Nine studies were included in the primary analysis of mortality, with a total of 1062 patients. None of the pooled RR's reached statistical significance. The pooled relative risk (RR) for mortality was 0.93 (95% confidence interval (CI) = 0.81 to 1.07). The overall RR for recovery of renal function, inclusive of 1038 patients from nine studies was 1.09 (95% CI 0.90 to 1.31). The pooled RR for mortality by dialyzer flux property was 1.03 (95% CI 0.82 to 1.30). The RR for recovery of renal function by flux property was 0.85 (95% CI 0.55 to 1.31). A meta-analysis of mortality of kidney transplant recipients was not possible, but the analysis of recovery of renal function in this patient population was 1.09 (95% CI 0.91to 1.31). Results of sensitivity analyses did not differ significantly from the primary analyses. AUTHORS' CONCLUSIONS: There is no demonstrable clinical advantage to the use of BCM versus BICM in patients with ARF who require intermittent hemodialysis.

Acute Kidney Injury↗

Mutation rates at Y chromosome specific microsatellites.

A collaborative work was carried out by the Spanish and Portuguese ISFG Working Group (GEP-ISFG) to estimate Y-STR mutation rates. Seventeen Y chromosome STR loci (DYS19, DYS385, DYS389I and II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS460, DYS461, DYS635 [GATA C4], GATA H4, and GATA A10) were analyzed in a sample of 3,026 father/son pairs. Among 27,029 allele transfers, 54 mutations were observed, with an overall mutation rate across the 17 loci of 1.998 x 10(-3) (95% CI, 1.501 x 10(-3) to 2.606 x 10(-3)). With just one exception, all of the mutations were single-step, and they were observed only once per gametogenesis. Repeat gains were more frequent than losses, longer alleles were found to be more mutable, and the mutation rate seemed to increase with the father's age. Hum Mutat 26(6), 520-528, 2005. (c) 2005 Wiley-Liss, Inc.

Age Factors↗

[Clinical practices of analgesia for invasive procedures in critically ill sedated patients in Ile-de-France: a phone survey].

To assess the practice of analgesia for invasive procedures in critically ill sedated patient in Ile-de-France (French area including Paris). Observational study: phone survey using a standard questionnaire. Only one senior physician in each of 30 intensive care unit (ICU) was questioned. Baseline sedation included systematic analgesia with narcotics in all ICUs. Only 4 physicians declared using a specific pain scale for sedated patients. Only 3 ICUs used written protocols. Procedures, which were thought to be most invasive (catheterization, pleural drainage, fibroscopy) were in most cases preceded by analgesia, but this was seldom the case for less painful events (venous or arterial puncture, tracheal suctioning). Specific pain scales are still underused. In contrast with current guidelines, analgesia for invasive procedures is not systematic but depends on subjective opinions.

Analgesia↗