Human islet isolation in fresh pancreas or after preservation in UW solution.
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Biomedical subjects
Publications and source records attributed to C Ramos.
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In the Valencian Community there are many areas in which the nitrate content of groundwater is higher than the limit of 50 mg l(-1) established by the European Union. A recent survey of drinking water quality showed that around 8% of the Valencian Community population had water supplies with nitrate content above that limit. The Valencian Community has an intensive agricultural base that includes citrus trees, fruit trees, and vegetables. A summary of the available nitrate leaching results in vegetable fields and citrus orchards is presented. It is demonstrated in the main vegetables grown, that N inputs were much higher than the values recommended by some researchers, and that nitrate leaching values were in most cases within the range of 150-300 kg N ha(-1). Artichoke, early potato, and onion were the three crops with higher leaching rates than other crops, based on simulation studies made for several of the main vegetable growing areas of the Valencian Community. The mineral N content in the 0-60 cm layer at planting time in the vegetable fields is, in many cases, greater than 200 kg ha(-1); fertilizer application could be greatly reduced if some recommendation system similar to the N(min) system used in other European countries was adopted. A simplified N balance in onion and early potato crops showed that the lower limit for mineralization in the 0-60 cm soil layer during the crop period varied from 65 to 130 kg N ha(-1). In these two crops nitrate leaching varied from 240 to 340 kg N ha(-1), depending on the N fertilizer treatment, and representing about 66-70% of total N input in the onion crop, and from 38 to 65% in the potato crop. Nitrate leaching losses in citrus orchards were, in general, lower than 100 kg N ha(-1) year(-1), representing about 33% of the total N input. A comparison of these results with those obtained in other studies is made. Finally, prospects for improvements in the fertilizer practice of vegetable crops and citrus orchards in the Valencian Community are discussed.
Aniridia can arise as part of the WAGR syndrome (Wilms tumour. aniridia, genitourinary anomalies, and mental retardation), due to a deletion or chromosomal region 11p13. We report a girl with a complete WAGR syndrome, whose brother presented hypospadias. Cytogenetic, FISH and molecular studies showed a deletion in one chromosome 11 of the patient. No cytogenetic rearrangement or deletion affecting the genes included in this region (PAX6 and WT1) were observed in her brother and parents. This excludes a higher risk than that of the general population for developing Wilms tumour in the brother and supports that the presence of WAGR syndrome in the patient and hypospadias in her brother is a chance association. We conclude that the identification and definition of the deletions in the WAGR region, which include the WT1 locus are important in order to identify a high tumour risk in infant patients with aniridia including those without other WAGR anomalies.
Objectives- Several attempts have been made to determine the gestational period in which the maximum number of fetal cells can be found in maternal blood and consequently which is the best week in which to perform a reliable non-invasive prenatal diagnosis. Most studies conclude that the number of nucleated red blood cells (NRBC) increases in line with gestation, but the number of cells that are fetal in origin (FNRBC) decreases in the third trimester. The aim of the present study was to make a practical comparative evaluation of the first and second trimesters to ascertain the period in which a greater number of FNRBC can be found of the total number of NRBC identified. Methods- Double density gradient and a posterior positive selection (CD71) by magnetic activated cell sorting (MACS) were employed. In the final fraction, erythroblasts were identified using Kleihauer staining and were studied using the fluorescence in situ hybridization (FISH) interphasic technique. Results- There was a significant difference (p<0.05) between the mean number of FNRBC found in the first and second trimesters. Conclusions- The number of FNRBC increases from the first to the second trimester. It appears that the optimum week in which to perform a reliable non-invasive prenatal diagnosis is around the 15th week.
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The objective of this paper is to describe the clinical and diagnostic characteristics of patients with pyogenic hepatic abscesses evaluated in the emergency room, and to know whether it is feasible to make an early diagnosis based on any of these characteristics. The setting was an urban, tertiary-care teaching hospital. This was a retrospective study of 63 adult patients admitted to our institution because of pyogenic liver abscesses from January 1991 to December 1998.
Several moderately halophilic gram-positive, spore-forming bacteria have been isolated by conventional enrichment cultures from damaged medieval wall paintings and building materials. Enrichment and isolation were monitored by denaturing gradient gel electrophoresis and fluorescent in situ hybridization. 16S ribosomal DNA analysis showed that the bacteria are most closely related to Halobacillus litoralis. DNA-DNA reassociation experiments identified the isolates as a population of hitherto unknown Halobacillus species.
Idiopathic pulmonary fibrosis (IPF) is a chronic lung disorder characterized by fibroblast proliferation and extracellular matrix accumulation. However, studies on fibroblast growth rate and collagen synthesis have given contradictory results. Here we analyzed fibroblast growth rate by a formazan-based chromogenic assay; fibroblast apoptosis by in situ end labeling (ISEL) and propidium iodide staining; percent of alpha-smooth muscle actin (alpha-SMA) positive cells by fluorescence-activated cell sorter; and alpha1-(I) collagen, transforming growth factor (TGF)-beta1, collagenase-1, gelatinases A and B, and tissue inhibitor of metalloproteinase (TIMP)-1, -2, -3, and -4 expression by reverse transcriptase/polymerase chain reaction in fibroblasts derived from IPF and control lungs. Growth rate was significantly lower in IPF fibroblasts compared with controls (13.3 +/- 38.5% versus 294.6 +/- 57%, P < 0.0001 at 13 d). Conversely, a significantly higher percentage of apoptotic cells was observed in IPF-derived fibroblasts (ISEL: 31.9 +/- 7.0% versus 15.5 +/- 7.6% from controls; P < 0.008). alpha-SMA analysis revealed a significantly higher percentage of myofibroblasts in IPF samples (62.8 +/- 25.2% versus 14.8 +/- 11.7% from controls; P < 0.01). IPF fibroblasts were characterized by an increase in pro-alpha1-(I) collagen, TGF-beta1, gelatinase B, and all TIMPs' gene expression, whereas collagenase-1 and gelatinase A expression showed no differences. These results suggest that fibroblasts from IPF exhibit a profibrotic secretory phenotype, with lower growth rate and increased spontaneous apoptosis.
Critical illness neuropathy frequently accompanies the septic syndrome in intensive care units. Another entity that may occur independently or concurrently in critically ill patients is a myopathy, giving rise to difficulties in distinguishing between them. The two patients described had sensorimotor axonal peripheral neuropathy. Axonopathy as a rule has a poor prognosis for recovery, which is slow and often incomplete. There are few reports of the functional outcome after rehabilitation therapy. The two patients described had 5-6 weeks of active rehabilitation therapy with improvement from moderate/severe disability at the onset to slight debility at the end of therapy.
Trypanosoma cruzi infection in central Mexico has not been fully documented, yet some data suggest its presence. In this work, sera from 211 subjects living in the state of Morelos and at risk of T. cruzi infection due to their living in contact with the vector were analyzed for the presence of antibodies to a total antigen extract of a Mexican isolate of T. cruzi. A seropositivity of 20% was demonstrated by both an enzyme-linked immunosorbent assay and Western blotting. Furthermore, parasites were isolated from five seropositive individuals, and these were genetically characterized as T. cruzi by multilocus enzyme electrophoresis. A case-control electrocardiographic study was conducted that included the seropositive individuals and twice as many seronegative controls living in the same area. A significant correlation was found between seropositivity and electrocardiographic alterations. These findings have important implications for perception of the prevalence of Chagas' disease in Mexico. Moreover, the presence of this disease in rural communities rapidly transforming into urban ones might have important epidemiologic consequences.
We report two infertile brothers presenting with azoospermia and oligozoospermia. Cytogenetic studies using G-banding and FISH analysis on lymphocyte cultures revealed an autosomal balanced reciprocal translocation t(1;22)(q11;p11) in both males. The same translocation was found in their mother, but not in a third fertile brother and maternal uncle suggesting that this translocation might compromise the male but not the female gametogenesis in this family.
We report on a girl with minor anomalies and developmental delay carrying an apparently balanced paracentric inversion of chromosome 6q (q22qter). Fluorescent in situ hybridization analysis demonstrated a deletion of the subtelomeric region of 6q. This illustrates the use of specific subtelomeric fluorescent in situ hybridization probes to detect cryptic deletions as an important cause of mental retardation in seemingly balanced chromosome rearrangements.
Recombinant derivatives of Pseudomonas putida strain KT2440 are of potential interest as microbial inoculants to be deliberately released for agricultural applications. To facilitate tracking of this strain and its derivatives after introduction into the environment, a mini-Tn5-'luxAB transposon was introduced into the chromosome of P. putida KT2440, yielding strain P. putida S1B1. Sequencing of the DNA region located upstream of the 'luxAB genes and similarity search with the P. putida KT2440 genome sequence, localized the transposon within a 3021-bp open reading frame (ORF), whose translated sequence showed significant similarity with the hypothetical YdiJ proteins from Escherichia coli and Haemophilus influenzae. A second ORF adjacent to and divergent from the ydiJ sequence was also found and showed significant homology with various LysR-type transcriptional activator proteins from several bacteria. Disruption of the ydiJ locus in P. putida S1B1 did not affect the survival of the strain in unvegetated or vegetated soils. Bioluminescent detection of P. putida S1B1 cells enriched in selective media directly from soil allowed detection of culturable cells in soil samples over a period of at least 8 months. The addition of the luxAB biomarker facilitates tracking in the root system of several plant species grown under sterile and non-sterile conditions. The correlation of the bioluminescent phenotype with the growth activity of P. putida S1B1 cells colonizing the root system of barley and corn plants was estimated by monitoring ribosomal contents using quantitative hybridization with fluorescence-labeled ribosomal RNA probes. A correlation between inoculum density, light output, and ribosomal contents was found for P. putida cells colonizing the root system of barley seedlings grown under sterile conditions. Although ribosomal contents, and therefore growth activity, of P. putida S1B1 cells extracted from the rhizosphere of corn plants grown in non-sterile soil were similar to those found in starved cells, the luminescent system permitted non-destructive in situ detection of the strain in the upper root system.
With the exception of a limited number of highly persistent lipophilic pollutants, waterborne exposure is considered the most relevant exposure route for aquatic organisms, and even the only relevant exposure route in lower tier assessment, while the potential for bioaccumulation is only evaluated as secondary poisoning for birds and mammals. In addition, some evaluations consider that only the dissolved fraction is bioavailable for pelagic organisms. The lack of easy and cost-effective assessment methods is probably the main reason to disregard the contribution of other routes. This paper presents a system dynamics model that estimates the time-dependent accumulation of toxic chemicals through the food chain, and has also the potential to include the exposure from sediment. The generic ecosystem includes a primary producer and three levels of consumers, represented by unicellular algae, cladoceran invertebrates, cladoceran-eating fish and fish-eating fish, respectively. Nevertheless, the model, built using Microsoft Excel software, allows any number of levels and ecological-exposure relationships. The results obtained for four illustrative pollutants demonstrate that factors such as the binding to algae and other edible particles or food chain exposure for lipophilic non-persistent chemicals may constitute significant exposure routes. The current low tier European ecological risk assessment guidelines do not cover these aspects, and therefore may under estimate the real risk for both aquatic organisms and their predators. The model includes a simplified worst-case alternative that normally can be calculated using the existing information. Under a tiered approach, the simplified alternative is proposed to trigger the need of the toxicokinetics assays required to perform the complete time-dependent calculation.
Three biochemical parameters, DNA quantification in soil samples and two enzymatic activities, beta-galactosidase and dehydrogenase have been assessed as potential end-points for the use in cost-effective toxicity tests on soil microorganisms. The assessment included the development of a classical dose-response 24-h assay and the incorporation of measurements of the effects on microbial activities in soil column leaching studies and multispecies miniaturised terrestrial systems (MTS). Four different chemicals, copper, a new herbicide, thiabendazole and fenthion were studied. A rapid fluorescence DNA quantification technique did not produce adequate responses. The efforts to quantify DNA after extraction and clean-up procedures failed due to the presence of humic acids. From the protocol of the technique one could see that the technical procedure is time-consuming and expensive and, for this reason, not suitable for use as a parameter in rapid and cost-effective tests. However, the enzymatic activities showed their potential as toxicity end-points. Copper produced a concentration/response inhibition of beta-galactosidase and dehydrogenase with EC50 values of 78.39 and 24.77 mg Cu/kg soil, respectively. In the soil column study, these endpoints allowed the measurement of the microbial activities through the column. The effects of the new herbicide on beta-galactosidase and dehydrogenase activities were statistically significant for the highest application dose (40 g/ha). Thiabendazole affected the microbial activity when mixed within the soil, but no effects were observed when this fungicide was applied on the soil surface. Fenthion produced effects when applied either in the soil or on the soil surface. These results can be explained by the low mobility of thiabendazole. The results show the capabilities of these biochemical parameters to be included as endpoints in cost-effective bioassays.
Hazard assessment constitutes an essential tool in order to evaluate the potential effects of chemical substances on organisms and ecosystems. It includes as a first step, hazard identification, which must detect the potential dangers of the substance (i.e. the kind of effects that the substance may produce), and a second step to quantify each danger and to set the expected dose/response relationships. Hazard assessment plays a key role in the regulation of chemical substances, including pollution control and sustainable development. However, the aquatic environment has largely received more attention than terrestrial ecosystems. This paper presents the extrapolation of several basic concepts from the aquatic to the terrestrial compartment, and suggests possibilities for their regulatory use. Two specific proposals are discussed. The first focuses on the scientific basis of the hazard identification-classification criteria included in the EU regulations and their extrapolation to the terrestrial environment. The second focuses on the OECD programme for environmental indicators and the development of a soil pollution pressure indicator to quantify the potential hazards for the soil compartment and its associated terrestrial ecosystem related to the toxic chemicals applied deliberately (i.e. pesticides) or not (i.e. heavy metals in sludge-based fertilisers; industrial spills) to the soil.
The risk assessment procedures of pesticides in the European Union are mostly based on worst-case scenarios developed for central and northern European conditions. From the point of view of Mediterranean conditions, these assessment procedures are not always appropriate, and therefore the development of specific scenarios is required. There are differences not only in meteorological data (temperature, solar irradiation, or rainfall volume and annual distribution) between these countries, but also in farm distribution, crop characteristics, soil or sediment properties and surface or groundwater characteristics. These differences are more evident for the typical crops of the southern countries, such as olive groves, vineyards, citrus or rice fields, and in these cases more realistic estimates of exposure are required. Our proposal includes the development of new criteria to improve the ecological risk assessment process for the conditions of the Mediterranean region, and the justification of this need.