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I Casas

Publications and source records attributed to I Casas.

At least 19 recordsLinked to original sources

Structural and ultrastructural features of boar seminal vesicles.

The morphological features of boar seminal vesicles were examined by light and transmission microscopy. Boar seminal vesicles consist of glandular tissue arranged in multiple lobules containing a system of ramified secretory tubules. The secretory tubules are composed of a mucosa formed by an epithelium and an underlying lamina propria and, are surrounded by a muscular layer. The epithelium is made up of columnar cells and occasional basal cells. Mast cells are frequently found among epithelial cells. Three types of columnar cells, considered different stages of the secretory cell cycle, are present: principal cells, clear cells and dense cells. Principal cells are functionally differentiated cells characterised by abundant mitochondria, great development of the rough endoplasmic reticulum and presence of secretory granules in their cytoplasm. The apical surface of many principal cells shows apical blebs filled with PAS-positive material. No acid mucosubstances are detected. Microvilli cover the apical surface except in the apical blebs. Dense cells, arranged between principal cells, are also functional differentiated cells but with signs of cellular degeneration. Clear cells are an initial differentiated stage of columnar cells and are characterised by the presence of a poorly developed rough endoplasmic reticulum and by the absence of secretory granules. Proliferating cells are present among columnar cells. Basal cells contain scarce cytoplasm, few organelles and no secretory granules. The lack of mitotic activity in these cells suggests that they do not act as precursors of columnar cells.

Animals↗

Human metapneumovirus infections in hospitalised infants in Spain.

BACKGROUND: Human metapneumovirus (hMPV) causes lower respiratory tract infections, particularly in young children and the elderly. METHODS: A prospective study was conducted on the clinical characteristics of infants <2 years of age admitted to hospital for respiratory infection and the characteristics of hMPV infections were compared with those of infections caused by respiratory syncytial virus (RSV). Influenza A, B and C viruses, RSV, parainfluenza viruses, and adenoviruses were simultaneously detected in clinical samples by multiple reverse transcription nested-PCR assay. The presence of hMPV was tested in all samples using two separate RT-PCR tests. RESULTS: A respiratory virus was detected in 65.9% of the 749 children included in the study. hMPV, found in 69 of the positive nasopharyngeal aspirates (14%), was the most common virus after RSV. Peak incidence was in March and over 80% of children were <12 months of age. The most common diagnoses were recurrent wheezing (49.3%) and bronchiolitis (46.4%). Oxygen therapy was required by 58% of patients, and assisted ventilation by one. Clinical characteristics in the 18 co-infections were indistinguishable from those of single infections. Fifty one hMPV single infections were compared with 88 hRSV single infections. Recurrent wheezing was diagnosed more frequently in hMPV patients. All other variables tested were similar in both groups. CONCLUSIONS: hMPV was the second most frequent virus after RSV in infants <2 years of age hospitalised for respiratory infection and was associated with lower respiratory tract infections. hMPV occurred predominantly in springtime. Co-infections were frequent and clinically similar to single infections and RSV infections.

Acute Disease↗

Prevalence and clinical characteristics of human metapneumovirus infections in hospitalized infants in Spain.

Human metapneumovirus (hMPV), a condition recently described in the Netherlands, causes lower respiratory infections, particularly in young children and among the elderly. The objective of this study was to describe the characteristics of hMPV infections in hospitalized infants <2 years of age and to compare them to those of infections caused by respiratory syncytial virus (RSV). A prospective study was conducted on the clinical characteristics of infants admitted to hospital for respiratory infection through 5 years. Simultaneous detection of influenza A, B, and C viruses, RSV, and adenoviruses was performed in clinical samples by multiple reverse transcription nested-PCR assay. The presence of hMPV was tested in all samples using two separate RT-PCR tests. Some respiratory virus was detected in 70.5% of the 1,322 children included in the study. hMPV was found in 101 of the positive nasopharyngeal aspirates (10.8%), and was the most common virus after RSV and rhinovirus. Peak incidence was found in March. Over 80% of children were <12 months. The more common diagnoses were bronchiolitis (49.5%) and recurrent wheezing (45.5%). Fifty-four percent of cases required oxygen therapy and, one percent, assisted ventilation. Thirty percent were co-infections, with clinical characteristics indistinguishable from single infections. Seventy-one hMPV single infections were compared to 88 RSV single infections. hMPV infections were significantly more frequent than RSV in infants older than 6 months (P = 0.04). Recurrent wheezing was diagnosed more frequently in hMPV patients (P = 0.001). All other variables tested were similar, in both groups. hMPV was the third most frequent virus after RSV and rhinovirus in infants <2 years of age, hospitalized for respiratory infection, and was associated with bronchiolitis and recurrent wheezing. hMPV predominantly occurred in spring. Co-infections were frequent and clinically similar to single infections and RSV infections.

Female↗

The use of a high-FeO olivine rock as a redox buffer in a nuclear waste repository.

Due to the higher stability of the spent nuclear fuel (mainly composed of UO2) under reducing conditions, and in order to enhance the retention/retardation of some key radionuclides, the olivine rock from the Lovasjärvi intrusion has been proposed as a potential redox-active backfill-additive in deep high-level nuclear waste (HLNW) repositories. In this work, two different approaches have been undertaken in order to establish the redox buffer capacity of olivine rock: (1) The capacity of the rock to respond to changes in pH or pe has been demonstrated and the final (pH, pe) coordinates agree with the control exerted by the system Fe(II)/Fe(III). (2) The rate of consumption of oxygen has been determined at different pH values. These rates are higher than the ones reported in the literature for other solids, what would point to the possibility of using this rock as an additive to the backfill material in a HLNW.

Buffers↗

Effect of culture conditions on the obtention of boar epididymal epithelial cell monolayers.

The goal of this study was to investigate the effect of the collagenase digestion time, the initial density of fragments and the culture temperature on the obtention of a boar epididymal epithelial cell culture, which is a useful methodology for the study of epididymal functions. A confluent monolayer of caput, corpus and cauda epididymal epithelial cells was only obtained when an adequate enzymatic digestion of the connective tissue surrounding the epididymal tubule was performed. For the correct digestion of caput and corpus fragments two collagenase digestions of 2 and 1h, respectively, were enough. Cauda fragments, however, needed two collagenase digestions of 3h each. A confluent monolayer of caput, corpus and cauda epididymal epithelial cells was obtained regardless of the initial density tested (15, 30, 60 and 90fragments/well). However, cultures originated from 15 and 30fragments/well showed higher cell concentration during the first 2 weeks of culture than cultures originated from 60 and 90fragments/well. A confluent monolayer of caput, corpus and cauda epididymal epithelial cells was obtained at both 32 and 37 degrees Celsius, but at 32 degrees Celsius cells grew very slowly and confluence was not reached until a week later than it was with cells growing at 37 degrees Celsius. In conclusion, we have observed that the time of digestion with collagenase is an important factor for the successful establishment of boar epididymal cell monolayers, and that the initial density of fragments and the culture temperature should be taken into account.

Animals↗

Structural and ultrastructural features of boar bulbourethral glands.

The morphological features of boar bulbourethral glands were examined by light and transmission microscopy. Bulbourethral glands are compound tubuloalveolar glands surrounded by a capsule of dense connective tissue and arranged in multiple lobules formed by endpieces and excretory ducts. Endpieces and excretory ducts are both lined by a single epithelium of mucous cells with a basal nucleus. Epithelial cells accumulate secretory granules containing neutral and carboxylated acid mucosubstances and a small amount of sulphated acid mucosubstances. The ultrastructure of epithelial cells varies according to the secretory cycle. In initial stages, the cells show a columnar shape and secretory granules unevenly distributed in the cytoplasm. As the synthesis of mucosubstances progresses, the amount of the secretory granules increases and the cellular shape becomes pyramidal. Secretory granules can contain inclusions and present differences among them according to their different phases of formation. In pyramidal cells, secretory products are released into the lumen by a merocrine mechanism.

Animals↗

Effects of exposing boars to different artificial light regimens on semen plasma markers and "in vivo" fertilizing capacity.

This study was designed to assess the effects of exposing boars to an artificial photoperiod on semen quality in terms of sperm concentration, sperm vitality, sperm motility and acrosome integrity. We also determined biochemical semen plasma variables, such as total protein concentration, phosphorylated tyrosine residues and fructose, glucose and sorbitol contents, along with their effects on the fertility, prolificacy and libido of the boars. Three groups of 10 males were kept for 3 months under experimental conditions of 24, 12 and 0 h of artificial light, and a constant temperature of 21 +/- 1 degrees C and 60-75% humidity. The animals were fed a nutritious diet and subjected to semen collection twice per week. Semen samples were analyzed throughout the entire experimental period. Our results indicate that, while the extreme photoperiods (0 and 24 h of light) affected semen quality in terms of sperm concentration, acrosome integrity and semen volume, its fertilizing capacity was only significantly reduced under conditions of absolute darkness. Sperm motility was found to be a poor indicator of fertilizing ability, while other sperm factors, such as acrosome integrity or other functional variables seemed to behave better. The photoperiod was found to affect the production of accessory sex gland secretions more than their composition. In addition, light effects on fertility, prolificacy and libido seemed to be achieved through independent mechanisms.

Acrosome↗

Evaluation of boar sperm maturation after co-incubation with caput, corpus and cauda epididymal cultures [corrected].

Boar sperm from the proximal caput epididymis were co-incubated with 1, 4, 7, 10 and 14-day old caput, corpus and cauda epididymal cultures for 24, 48 and 72 h. Boar kidney epithelial cells (LLC-PK1) and ECM alone were used as negative controls. Sperm motility, morphology and membrane integrity were studied to evaluate boar sperm maturation in vitro. Our results showed that epithelial cell monolayers (10, 14-day old) create a suitable microenvironment for the survival of proximal caput sperm and the maintenance of sperm motility over a 72 h period. Moreover, corpus epididymal tubule fragments in culture (1, 4-day old) are capable of promoting the migration of the cytoplasmic droplet along the sperm tail after 24h of co-incubation.

Animals↗

Molecular identification of adenoviruses in clinical samples by analyzing a partial hexon genomic region.

Here we present a system for adenovirus detection and genotyping based on PCR amplification and phylogenetic analysis of a conserved hexon gene fragment. The system was validated using 157 sequences (86 previously typed and 71 clinical samples) and correctly identified species and serotype in 100% and 84% of sequences, respectively. Known associations between specific serotypes and clinical syndromes are verified. Possible new associations are described to allow further independent testing.

Adenovirus Infections, Human↗

Formation of studtite during the oxidative dissolution of UO2 by hydrogen peroxide: a SFM study.

Understanding the formation of alteration phases on the surface of spent nuclear fuel, such as those observed during leaching experiments, is necessary in order to predict the concentration of radionuclides in the near-field of a final repository. Hydrogen peroxide has been identified as one of the oxidants formed by the radiolysis of water in the presence of spent nuclear fuel; especially due to alpha activity. The presence of this species in solution can contribute to the formation of uranium peroxide secondary phases. In this work, we have studied the oxidative dissolution of synthetic UO2 disks in hydrogen peroxide solutions of two different concentrations (5 x 10(-4) and 5 x 10(-6) mol dm(-3)), both at pH 5.8 +/- 0.1. The solid surface evolution of the disks has been followed by means of ex-situ scanning force microscope (SFM) measurements, and uranium concentration in solution has been determined by inductively coupled plasma mass spectrometry. During the first stage of the experiment, SFM images indicate that only UO2 dissolution is occurring. After 142 h, a secondary phase is observed on the surface of the solid at 5 x 10(-4) mol dm(-3) hydrogen peroxide concentration. This secondary phase has been identified by X-ray diffraction as studtite (UO4 x 4H2O). From the analysis of SFM topographic profiles at different elapsed times, a precipitation rate for the studtite has been estimated to be in the range of (8-32) x 10(-10) mol m(-2) s(-1).

Forecasting↗

Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays.

There is a need for rapid, sensitive, and accurate diagnosis of lower respiratory tract infections in children, elderly, and immunocompromised patients, who are susceptible to serious complications. The multiplex RT-nested PCR assay has been used widely for simultaneous detection of non-related viruses involved in infectious diseases because of its high specificity and sensitivity. A new multiplex RT-PCR assay is described in this report. This approach includes nested primer sets targeted to conserve regions of human parainfluenza virus haemagglutinin, human coronavirus spike protein, and human enterovirus and rhinovirus polyprotein genes. It permits rapid, sensitive, and simultaneous detection and typing of the four types of parainfluenza viruses (1, 2, 3, 4AB), human coronavirus 229E and OC43, and the generic detection of enteroviruses and rhinoviruses. The testing of 201 clinical specimens with this multiplex assay along with other one formerly described by our group to simultaneously detect and type the influenza viruses, respiratory syncytial viruses, and a generic detection of all serotypes of adenovirus, covers the detection of most viruses causing respiratory infectious disease in humans. The results obtained were compared with conventional viral culture, immunofluorescence assay, and a third multiplex RT-PCR assay for all human parainfluenza viruses types described previously. In conclusion, both multiplex RT-PCR assays provide a system capable of detecting and identifying simultaneously 14 different respiratory viruses in clinical specimens with high sensitivity and specificity, being useful for routine diagnosis and survey of these viruses within the population.

Coronavirus↗

An outbreak of adenovirus type 8 keratoconjunctivitis in a nursing home in Madrid.

This work describes and analyses an outbreak of epidemic keratoconjunctivitis which occurred in 2001 and 2002 in a nursing home for the elderly in Leganes (an area of Madrid). This is the first such published case in Spain with these characteristics and this serotype identification. Sociodemographic characteristics, epidemic curve and attack rates are described. Comparisons of the data were carried out using a chi2 test for qualitative variable and t-test for quantitative. Factors associated with the illness are explored by means of contingency tables and logistic regression models. One hundred and two cases were detected, with an attack rate of 36.4% for residents, and 12.9% for workers, not considering spatial or professional differences. The epidemic curve showed an interpersonal transmission pattern. Multivariate analysis identified the following risk factors in the residents: able to wander freely through the building, urinary incontinence and use of shared bathroom. In 34.6% of the conjunctival samples, adenovirus serotype 8 was detected with identical genomic sequence. Establishment of hygienic sanitary guidance adapted for the cleaning of such establishments and contact with residents as well as early diagnosis and good coordination of human and material resources are key factors in the prevention and control of these outbreaks in closed communities.

Adenovirus Infections, Human↗

Simultaneous detection of influenza A, B, and C viruses, respiratory syncytial virus, and adenoviruses in clinical samples by multiplex reverse transcription nested-PCR assay.

The clinical presentation of infections caused by the heterogeneous group of the respiratory viruses can be very similar. Thus, the implementation of virological assays that rapidly identify the most important viruses involved is of great interest. A new multiplex reverse transcription nested-polymerase chain reaction (RT-PCR) assay that is able to detect and type different respiratory viruses simultaneously is described. Primer sets were targeted to conserved regions of nucleoprotein genes of the influenza viruses, fusion protein genes of respiratory syncytial viruses (RSV), and hexon protein genes of adenoviruses. Individual influenza A, B, and C viruses, RSV (A and B), and a generic detection of the 48 serotypes of adenoviruses were identified and differentiated by the size of the PCR products. An internal amplification control was included in the reaction mixture to exclude false-negative results due to sample inhibitors and/or extraction failure. Detection levels of 0.1 and 0.01 TCID50 of influenza A and B viruses and 1-10 molecules of cloned amplified products of influenza C virus, RSV A and B, and adenovirus serotype 1 were achieved. The specificity was checked using specimens containing other respiratory viruses and no amplified products were detected in any case. A panel of 290 respiratory specimens from the 1999-2000 and 2000-2001 seasons was used to validate the assay. Accurately amplifying RNA from influenza and RSV prototype strains and DNA from all adenovirus serotypes demonstrates the use of this method for both laboratory routine diagnosis and surveillance of all these viruses.

Adenoviruses, Human↗

First epidemic of aseptic meningitis due to echovirus type 13 among Spanish children.

Echoviruses are the commonest cause of aseptic meningitis (AM). Echovirus type 13 (EV-13) was the second enterovirus serotype associated with different local outbreaks of AM in Spain between February and October 2000. It was the first time that an epidemic AM caused by this virus was recognized in Spain. The index case appeared in the Canary Islands (Canarias). The EV-13 virus was isolated from 135 patients, predominantly from cerebrospinal fluid (CSF). All isolates were from children under 13 years. The age specific peak incidence was in infants under 1 year. Most patients had fever, headache and other meningeal signs. This enterovirus serotype, not previously detected in Spain, caused severe illness with a high attack rate.

Adolescent↗

Molecular identification of enterovirus by analyzing a partial VP1 genomic region with different methods.

VP1 is the most suitable region for use in the identification of enterovirus. Although VP1 sequencing methods may vary, it is necessary to agree on a common strategy of sequence analysis. Identification of a strain type may be achieved by three different approaches: pairwise sequence alignment, multiple-sequence alignment, and phylogenetic inference. Other methods are also available, but they are not simple enough to be performed at a virology laboratory. The performances of these methods were evaluated with nucleotide and protein sequences obtained from 32 original samples, 8 enterovirus isolates, and 64 GenBank sequences. Pairwise sequence alignment methods had very different results. The DNASTAR package identified only 28.8% of enterovirus strains, while the Genetics Computer Group package identified 50.0 or 72.1% of enterovirus strains when nucleotide or amino acid sequences were analyzed, respectively. Multiple-sequence alignment methods identified 94.2% (Clustal W program) or 92.3% (Pileup program) of the enterovirus strains, while the phylogenetic method increased this rate to 99.0%. Comparative evaluation of these analysis methods showed that the Clustal W program (version 1.81), a freely available multiple-sequence alignment program, presented one of the best performances when used with the correct criteria. Other commercial and expensive programs did not achieve the same performances, making them less suitable for molecular typing of enteroviruses. Finally, although phylogenetic inference is the most demanding method in terms of knowledge of the user, it remained the best option analyzed.

Algorithms↗

Molecular epidemiology of echovirus 30: temporal circulation and prevalence of single lineages.

Echovirus 30 (EV30) is one of the most frequently isolated EVs, causing extensive outbreaks of EV30 aseptic meningitis in temperate climates. EV30 is antigenically heterogeneous, and three major antigenic groups have been defined, although the basis for the antigenic differences is unknown. A reverse transcription-nested PCR which amplifies the 3'-terminal region of the VP1 gene directly from clinical samples was selected for studying EV30 molecular epidemiology, since the major antigenic sites in this region reflect the serotypic pattern of this virus. The different previous approaches to the genetic classification of EV30 were analyzed. A complete study of the EV30 strains was performed by analyzing the sequences from the 112 EV30 strains amplified in this work and the complete set of EV30 strains previously published. A total of 318 strains of EV30 were divided into two broad genotypes (I and II). This classification was supported by the phylogenetic trees obtained from amino acid sequences, and it correlated with the antigenic heterogeneity of the reference strains described in earlier studies. The genotypes could be further divided into subgroups, and these subgroups could be divided into lineages based on their nucleotide distances and levels of bootstrapping. On the other hand, the subgroups and lineages did not result in the same correlation between amino acid and nucleotide differentiation. The molecular epidemiology of EV30 can be compared to influenza virus epidemiology, where prevailing lineages displace the less established lineages on the basis of immune escape. This pattern of evolution is clearly different from that of other enteroviruses. A single lineage at a time appears to be circulating worldwide. This behavior may be related to the epidemic activity of EV30.

Argentina↗

Molecular characterization of human enteroviruses in clinical samples: comparison between VP2, VP1, and RNA polymerase regions using RT nested PCR assays and direct sequencing of products.

Three nested RT-PCR assays were developed to permit sensitive typing of enteroviruses directly from clinical samples. These assays amplified short fragments from different genomic regions codifying for three proteins: VP2, VP1, and RNA polymerase. Given that enteroviruses have a high rate of degeneration within target codons among serotypes, the primers used consisted of mixed base and deoxyinosine residues. These techniques detected at 0.03-0.003 TCID50 of prototype Poliovirus 1 and Echovirus 30. They were used to characterize the enteroviral RNA detected in 18 CSF, stool, and throw swab samples and in 8 enterovirus isolates from patients with several syndromes. Phylogenetic analysis in each independent sequenced region grouped the enterovirus into four clusters, enabling genetic classification. A comparative study was performed among the 26 sequences obtained after direct sequencing of products with those available in the nucleotide databases. The efficiency of each assay for enterovirus identification was evaluated by both distance (Clustal) and similarity (M-NW) indices. Comparative results obtained independently in the three regions showed the highest yield of correlation between nucleotide sequences of all prototype serotypes and the analyzed genotypes in the VP1 region (26/26, 100% Clustal; 22/26, 85% M-NW). Conversely, the VP2 region failed to identify some of the circulating enteroviruses (17/26, 65% Clustal; 16/26, 62% M-NW). Using the RNA polymerase region, sequences from samples and isolates were associated with prototype strains whenever these were available (20/21, 95% Clustal; 12/21, 57% M-NW). These assays were useful for molecular identification of enterovirus directly from samples even when isolation was not possible.

Capsid↗