Comparative Expression Profiling of the Chlamydia Trachomatis Pmp Gene Family for Clinical and Reference Strains

PloS One. 2007  |  Pubmed ID: 17849007

Chlamydia trachomatis, an obligate intracellular pathogen, is a leading worldwide cause of ocular and urogenital diseases. Advances have been made in our understanding of the nine-member polymorphic membrane protein (Pmp) gene (pmp) family of C. trachomatis. However, there is only limited information on their biologic role, especially for biological variants (biovar) and clinical strains.

Streptococcus Agalactiae Serotype Ib As an Agent of Meningitis in Two Adult Nonpregnant Women

Journal of Clinical Microbiology. Nov, 2007  |  Pubmed ID: 17881554

Two temporally and geographically clustered cases of meningitis caused by Streptococcus agalactiae expressing the infrequent Ib serotype are reported. Characterization by pulsed-field gel electrophoresis and multilocus sequence typing revealed that the isolates were identical and represented the widely distributed ST10/ST8 lineage associated with serotype Ib.

Association Study of 69 Genes in the Ret Pathway Identifies Low-penetrance Loci in Sporadic Medullary Thyroid Carcinoma

Cancer Research. Oct, 2007  |  Pubmed ID: 17909067

To date, few association studies have been done to better understand the genetic basis for the development of sporadic medullary thyroid carcinoma (sMTC). To identify additional low-penetrance genes, we have done a two-stage case-control study in two European populations using high-throughput genotyping. We selected 417 single nucleotide polymorphisms (SNP) belonging to 69 genes either related to RET signaling pathway/functions or involved in key processes for cancer development. TagSNPs and functional variants were included where possible. These SNPs were initially studied in the largest known series of sMTC cases (n = 266) and controls (n = 422), all of Spanish origin. In stage II, an independent British series of 155 sMTC patients and 531 controls was included to validate the previous results. Associations were assessed by an exhaustive analysis of individual SNPs but also considering gene- and linkage disequilibrium-based haplotypes. This strategy allowed us to identify seven low-penetrance genes, six of them (STAT1, AURKA, BCL2, CDKN2B, CDK6, and COMT) consistently associated with sMTC risk in the two case-control series and a seventh (HRAS) with individual SNPs and haplotypes associated with sMTC in the Spanish data set. The potential role of CDKN2B was confirmed by a functional assay showing a role of a SNP (rs7044859) in the promoter region in altering the binding of the transcription factor HNF1. These results highlight the utility of association studies using homogeneous series of cases for better understanding complex diseases.

[Preliminary Study on Efficacy and Tolerance of a "coupage" of Olive Oil in Patients with Chronic Kidney Disease. Nutritonal Evaluation]

Nefrología : Publicación Oficial De La Sociedad Española Nefrologia. 2007  |  Pubmed ID: 17944585

The discrepancies among data reported by using olive oil (OO) in humans appear to be due to the great differences between the different OO used. Based on structure/function relationships we have chemically optimized an OO through the rational mixture ("coupage") of several Spanish extra virgin olive oils (methodology "oHo"). Patients with chronic kidney disease (CKD) develop a progressive picture of malnutrition and inflammation that lead them to an elevated risk of cardiovascular disease. In a pilot, randomised trial the nutritional efficacy and safety of "oHo" were evaluated in 32 patients (mean age 60,8 +/- 13,2 years old; 16 women) with CKD (KDIGO stages 4-5) at predialysis. After a 7 days wash out for statins and ACE inhibitors 19 patients had "oHo" at doses of 60 mL/day (20 mL t.i.d) for 30 consecutive days, whilst 13 patients remain as a control group without "oHo". At the end of the study only patients having "oHo" showed significant increases of serum albumin (p<0.05) and not significant increases of total proteins, weight, and BMI. Total cholesterol (p<0.05) and HDL-cholesterol (p<0.01) increased with "oHo". The number of cases with pathologic HOMA-IR in the control group increased from 1 to 2 patients whilst in the "oHo" group decreased from 2 to none. No significant changes of minerals, arterial pressure, hemoglobin, and other parameters related to CKD were seen. After a 30 days follow-up in the "oHo" group all parameters came back to basal ones, excepting for blood pressure that significantly decreased (p<0,05). Tolerance was excellent and constipation significantly diminished (p<0,001) in the "oHo" group. Of importance, none of these biological changes were seen in regular consumers of other conventional olive oils (control group). These intriguing results, seen by the first time, appear to partially satisfy the recent claims ("reverse epidemiology") about the need of a more correct nutrition in CKD patients. However, these data need to be proved in more larger trials as well as in CKD patients under dialysis with harder inflammatory/malnutrition conditions.

Pharmacy Students' Knowledge, Attitudes, and Evaluation of Direct-to-consumer Advertising

American Journal of Pharmaceutical Education. Oct, 2007  |  Pubmed ID: 17998983

To assess pharmacy students' knowledge, attitudes, and evaluation of direct-to-consumer advertising (DTCA).

Co-occurrence of Viral and Bacterial Pathogens in Disease Outbreaks Affecting Newly Cultured Sparid Fish

International Microbiology : the Official Journal of the Spanish Society for Microbiology. Sep, 2007  |  Pubmed ID: 18076001

Several microbial disease outbreaks in farm stocks of newly cultured sparid fish species, such as common seabream, redbanded seabream, and white seabream, were recorded from 2004 to 2006. This study describes the isolation and characterization of the potential causative agents, either bacteria or viruses, of these outbreaks. The isolated bacterial strains were characterized according to traditional taxonomical analyses and sequencing of a 16S rDNA fragment. Most bacteria were identified as Vibrio spp. and Photobacterium damselae subsp. damselae. The development of cytopathic effects (CPE) on different fish cell lines, the application of specific nested-PCR tests for infectious pancreatic necrosis virus (IPNV), viral nervous necrosis virus (VNNV) and viral hemorrhagic septicemia virus (VHSV), and subsequent sequence analyses were used for virus detection and identification. VNNV, related to the striped jack neural necrosis virus (SJNNV) genotype, and VHSV, related to the genotype Ia, were the only viruses detected. VNNV was isolated from the three fish species under study in five different outbreaks, whereas VHSV was isolated from common seabream and white seabream during two of these outbreaks. IPNV was not detected in any case.

Congenital Melanocytic Nevi: Treatment Modalities and Management Options

Seminars in Cutaneous Medicine and Surgery. Dec, 2007  |  Pubmed ID: 18395671

Congenital melanocytic nevi can be cosmetically disfiguring, give rise to melanoma, and suggest the presence of neurocutaneous melanocytosis. Management decisions must be tailored for each patient and each nevus, taking into consideration the risk for developing malignancy, risk for developing symptomatic neurocutaneous melanocytosis, cosmetic implications of having the nevus, cosmetic implications of any resultant surgical scars from their removal, adverse effects that the nevus may have on psycho-social development, and the adverse effects and long-term sequelae of any surgical intervention. The advantages and disadvantages of different modalities used in the treatment of congenital melanocytic nevi are discussed. Organizational flow diagrams are presented to help clinicians in managing patients with different sized congenital melanocytic nevi.

The Heterodimeric Assembly of the CD94-NKG2 Receptor Family and Implications for Human Leukocyte Antigen-E Recognition

Immunity. Dec, 2007  |  Pubmed ID: 18083576

The CD94-NKG2 receptor family that regulates NK and T cells is unique among the lectin-like receptors encoded within the natural killer cell complex. The function of the CD94-NKG2 receptors is dictated by the pairing of the invariant CD94 polypeptide with specific NKG2 isoforms to form a family of functionally distinct heterodimeric receptors. However, the structural basis for this selective pairing and how they interact with their ligand, HLA-E, is unknown. We describe the 2.5 A resolution crystal structure of CD94-NKG2A in which the mode of dimerization contrasts with that of other homodimeric NK receptors. Despite structural homology between the CD94 and NKG2A subunits, the dimer interface is asymmetric, thereby providing a structural basis for the preferred heterodimeric assembly. Structure-based sequence comparisons of other CD94-NKG2 family members, combined with extensive mutagenesis studies on HLA-E and CD94-NKG2A, allows a model of the interaction between CD94-NKG2A and HLA-E to be established, in which the invariant CD94 chain plays a more dominant role in interacting with HLA-E in comparison to the variable NKG2 chain.

[Psychosis and Substance Abuse]

Acta Médica Portuguesa. Sep-Oct, 2007  |  Pubmed ID: 18282438

The objective of this retrospective study covering a period of 4 years (2001-2004), is to characterize and chart the clinical profiles of acute patients in the psychiatric inpatients ward of Hospital S. Francisco Xavier diagnosed with psychosis and substance abuse. During this period, 422 patients diagnosed with nonaffective psychosis (292, 295, 297 and 298, according to ICD9) were admitted. An analysis of the clinical profiles of these inpatients shows that a high percentage (42%) have psychotic pathologies and an history of substance abuse. These are predominantly young males. Schizophrenic patients with an history of substance abuse reached 40%. A particularly relevant finding in view of the fact that substance abuse leads to worse prognostics. Cannabinoids are by far the most widely abused substance, followed by alcohol. There are also 7.2% psychosis due to drug use in which the drug of choice was cannabis. In the light of current scientific knowledge, this is very relevant fact since the use of cannabinoids appears to be a risk factor for the onset of schizophrenia. It was observed that, often, due importance is not given to this association since few nodrug assays are carried out and when they are, they don't cover all drugs. Therefore, this study should alert medical staff to the high prevalence of substance abuse amongst psychotic patients, and to the necessity for a proactive stance in the detection of substance abuse in order to facilitate the development of better therapeutic projects which take into consideration this comorbidity and seek to promote psychoeducation of patients.

The Clinical Use of Exhaled Nitric Oxide in Wheezing Children

Revista Portuguesa De Pneumologia. Mar-Apr, 2008  |  Pubmed ID: 18363018

The body of published work on the role of exhaled nitric oxide (FENO) in the study of bronchial inflammation allows it to be classed as a simple, non-invasive measurement that is very useful in evaluating asthmatic patients. During a prospective study into the effects of air pollution on the health of the population of Viseu (Saud'AR Project), children with a clinical history of wheezing were identified through using the International Study of Asthma and Allergy in Childhood (ISAAC) questionnaire. Children later filled in a new standardised questionnaire and underwent skin-prick-tests, spirometry and FENO measurement. Their mean age was 7.8+/-1.1 years. Comparing those who wheezed in the 6 months before evaluation (n=27) with those who didn't, statistical differences for DeltaFEV1 (8% median versus 4.5%, p=0.0399) and for FENO (23 ppb median versus 12 ppb, p=0.0195, respectively) were observed. Concerning children who needed a bronchodilator in the six previous months (n=19) and those who didn't, there was also a statistically significant difference in FENO: 27 ppb median versus 11 ppb median, respectively; p<0.0001. When comparing children who needed an unscheduled medical appointment in the six months previous to the evaluation (n=9) and those who didn't, there was also significant differences for FE NO: 28 ppb median versus 13 ppb median, p=0.0029. In conclusion, the existence of symptoms seems to be better related to FE NO than spirometry.

Uncommon Endocytic and Trafficking Pathway of the Natural Killer Cell CD94/NKG2A Inhibitory Receptor

Traffic (Copenhagen, Denmark). Jun, 2008  |  Pubmed ID: 18363778

The CD94/NKG2A inhibitory receptor, expressed by natural killer and T cells, is constantly exposed to its HLA-E ligand expressed by surrounding cells. Ligand exposure often induces receptor downregulation. For CD94/NKG2A, this could potentiate activation receptor(s) induced responses to normal bystander cells. We investigated CD94/NKG2A endocytosis and found that it occurs by an amiloride-sensitive, Rac1-dependent macropinocytic-like process; however, it does not require clathrin, dynamin, ADP ribosylation factor-6, phosphoinositide-3 kinase or the actin cytoskeleton. Once endocytosed, CD94/NKG2A traffics to early endosomal antigen 1(+), Rab5(+) early endosomes. It does appear in Rab4(+) early/sorting endosome, but, in the time period examined, fails to reach Rab11(+) recycling or Rab7(+) late endosomes or lysosome-associated membrane protein-1(+) lysosomes. These results indicate that CD94/NKG2A utilizes a previously undescribed endocytic mechanism coupled with an abbreviated trafficking pattern, perhaps to insure surface expression.

EYS, Encoding an Ortholog of Drosophila Spacemaker, is Mutated in Autosomal Recessive Retinitis Pigmentosa

Nature Genetics. Nov, 2008  |  Pubmed ID: 18836446

Using a positional cloning approach supported by comparative genomics, we have identified a previously unreported gene, EYS, at the RP25 locus on chromosome 6q12 commonly mutated in autosomal recessive retinitis pigmentosa. Spanning over 2 Mb, this is the largest eye-specific gene identified so far. EYS is independently disrupted in four other mammalian lineages, including that of rodents, but is well conserved from Drosophila to man and is likely to have a role in the modeling of retinal architecture.

Regulation of Human DAP10 Gene Expression in NK and T Cells by Ap-1 Transcription Factors

Journal of Immunology (Baltimore, Md. : 1950). Jan, 2008  |  Pubmed ID: 18097042

Human NKG2D/DAP10 is an activation receptor expressed by NK and subsets of T cells, whose ligands include MHC class I chain-related (MIC) protein A and protein B and UL16-binding proteins that are often up-regulated by stress or pathological conditions. DAP10 is required for NKG2D/DAP10 cell surface expression and signaling capacity. Little is known about the mechanisms that regulate DAP10 gene expression. We describe the existence of multiple transcriptional start sites upstream of DAP10 exon 1 and identify the location of the basic promoter upstream of these starting sites. The promoter is active in NK and CD8+ T cells, but not in CD4+ T cells. We demonstrate TCR-mediated up-regulation of DAP10 transcription and found that a 40 bp region within the DAP10 promoter, containing an Ap-1 binding site, is largely responsible for this increased transcription. Using pull-down and chromatin immunoprecipitation assays, we show that the DAP10 promoter interacts with Ap-1 transcription factors in primary CD8+ T and NK cells in vitro and in vivo. Overexpression of c-Jun or c-Fos in NK and T cells led to enhanced DAP10 promoter activity and DAP10 protein expression. Taken together, our data indicate that Ap-1 is an important transcription factor for regulating DAP10 gene expression in human NK and T cells, and that Ap-1 plays a key role in the transactivation of DAP10 promoter following TCR stimulation.

Poly I:C Induces Mx Transcription and Promotes an Antiviral State Against Sole Aquabirnavirus in the Flatfish Senegalese Sole (Solea Senegalensis Kaup)

Fish & Shellfish Immunology. Mar, 2008  |  Pubmed ID: 18191581

Mx is an interferon-induced protein that protects against viral infections. In this study the absolute number of Mx transcripts after poly I:C injection (a synthetic dsRNA) or sole aquabirnavirus (solevirus) inoculation in Senegalese sole (Solea senegalensis Kaup) has been quantified. Mx expression profiles differed clearly in both experimental conditions; the induction response was faster and more intense after poly I:C injection than after solevirus inoculation. Moreover, pre-injection of soles with poly I:C prior to solevirus infection eliminated the induction of Mx expression associated with this virus. To evaluate the possible interference of poly I:C treatments on solevirus replication, the mRNA levels of the virus capsid protein (VP2) were determined by RT-PCR. VP2 transcripts were hardly detected in poly I:C pre-injected animals from 12 to 72 h after solevirus inoculation. All these data suggest that poly I:C is able to induce an antiviral state that interferes with solevirus replication, and support the suitability of Mx expression analysis as a marker to study the defensive response against solevirus.

The NKG2D Receptor: Immunobiology and Clinical Implications

Immunologic Research. 2008  |  Pubmed ID: 18193361

NK cells are critical components of our immune system functioning, in part, to recognize and then eradicate virally infected or tumorigenic cells without previous sensitization. One of the best-characterized activating receptors expressed on NK cells is the NKG2D receptor, which is capable of transmitting co-stimulatory signals by subsets of T cells. Viruses and tumors have evolved strategies to evade NKG2D-mediated immune recognition thus highlighting the importance of this receptor in immunity. This review will focus on the structure of NKG2D and its interaction with its diverse array of ligands, as well as highlighting current knowledge regarding NKG2D signal transduction and biological mechanisms that govern its cell surface expression. The impact that NKG2D has in disease pathologies is also assessed.

Procedures for Estimation of Modelling Uncertainty in Air Quality Assessment

Environment International. Jul, 2008  |  Pubmed ID: 18234341

The main objectives of this work focus, firstly, on a review of the current existent methodologies to estimate air quality modelling uncertainty, and, secondly, in the preparation of guidelines for modelling uncertainty estimation, which can be used by local and regional authorities responsible for air quality management. From the application exercise, it was concluded that it is possible to define a subset of statistical parameters able to reproduce the general uncertainties estimation. Concerning the quality indicators defined by EU directives, the results show that the legislated uncertainty estimation measures are ambiguous and inadequate in several aspects, mainly in what concerns the error measures for hourly and daily indicators based on the highest observed concentration. A relative error at the percentile correspondent to the allowed number of exceedances of the limit value was suggested and tested, showing that is a more robust and appropriate parameter for model performance evaluation.

Subcellular Distribution of Swine Vesicular Disease Virus Proteins and Alterations Induced in Infected Cells: a Comparative Study with Foot-and-mouth Disease Virus and Vesicular Stomatitis Virus

Virology. May, 2008  |  Pubmed ID: 18279902

The intracellular distribution of swine vesicular disease virus (SVDV) proteins and the induced reorganization of endomembranes in IBRS-2 cells were analyzed. Fluorescence to new SVDV capsids appeared first upon infection, concentrated in perinuclear circular structures and colocalized to dsRNA. As in foot-and-mouth disease virus (FMDV)-infected cells, a vesicular pattern was predominantly found in later stages of SVDV capsid morphogenesis that colocalized with those of non-structural proteins 2C, 2BC and 3A. These results suggest that assembly of capsid proteins is associated to the replication complex. Confocal microscopy showed a decreased fluorescence to ER markers (calreticulin and protein disulfide isomerase), and disorganization of cis-Golgi gp74 and trans-Golgi caveolin-1 markers in SVDV- and FMDV-, but not in vesicular stomatitis virus (VSV)-infected cells. Electron microscopy of SVDV-infected cells at an early stage of infection revealed fragmented ER cisternae with expanded lumen and accumulation of large Golgi vesicles, suggesting alterations of vesicle traffic through Golgi compartments. At this early stage, FMDV induced different patterns of ER fragmentation and Golgi alterations. At later stages of SVDV cytopathology, cells showed a completely vacuolated cytoplasm containing vesicles of different sizes. Cell treatment with brefeldin A, which disrupts the Golgi complex, reduced SVDV (approximately 5 log) and VSV (approximately 4 log) titers, but did not affect FMDV growth. Thus, three viruses, which share target tissues and clinical signs in natural hosts, induce different intracellular effects in cultured cells.

Envelope-specific Antibody Response in HIV-2 Infection: C2V3C3-specific IgG Response is Associated with Disease Progression

AIDS (London, England). Nov, 2008  |  Pubmed ID: 18981765

To examine the unspecific and envelope-specific IgA and IgG responses in acute and chronic HIV-2 infection.

Bilateral Areolar Sebaceous Hyperplasia in a Female

International Journal of Dermatology. Nov, 2008  |  Pubmed ID: 18986471

Chlamydia Trachomatis Diversity Viewed As a Tissue-specific Coevolutionary Arms Race

Genome Biology. 2008  |  Pubmed ID: 18947394

The genomes of pathogens are thought to have evolved under selective pressure provided by the host in a coevolutionary arms race (the 'Red Queen's Hypothesis'). Traditionally, adaptation by pathogens is thought to rely not on whole chromosome dynamics but on gain/loss of specific genes, yielding differential abilities to infect distinct tissues. Thus, it is not known whether distinct host organs differently shape the genome of the same pathogen. We tested this hypothesis using Chlamydia trachomatis as model species, looking at 15 serovars that infect different organs: eyes, genitalia and lymph nodes.

Polymorphisms in the Genes Encoding the 4 RET Ligands, GDNF, NTN, ARTN, PSPN, and Susceptibility to Hirschsprung Disease

Journal of Pediatric Surgery. Nov, 2008  |  Pubmed ID: 18970938

Hirschsprung disease (HSCR) is a developmental disorder caused by a failure of neural crest cells to migrate, proliferate, and/or differentiate during the enteric nervous system development. It presents a multifactorial, nonmendelian pattern of inheritance, with several genes playing some role in its pathogenesis. Its major susceptibility gene is the RET protooncogene, which encodes a receptor tyrosine kinase activating several key signaling pathways in the enteric nervous system development. Given the pivotal role of RET in HSCR, the genes encoding their ligands (GDNF, NRTN, ARTN, and PSPN) are also good candidates for the disease.

Depressive Symptoms in Patients with Type 2 Diabetes in the Ambulatory Care Setting: Opportunities to Improve Outcomes in the Course of Routine Care

Journal of the American Pharmacists Association : JAPhA. Nov-Dec, 2008  |  Pubmed ID: 19019802

To assess the frequency of untreated, self-reported depressive symptoms in a cross section of adult ambulatory patients with type 2 diabetes and to identify demographic and/or clinical characteristics associated with depressive symptoms in study patients.

Transcriptional Expression of Cis-acting and Trans-acting Splicing Mutations Cause Autosomal Dominant Retinitis Pigmentosa

Human Mutation. Jun, 2008  |  Pubmed ID: 18412284

Two types of mutations may lead to deficient pre-mRNA splicing: cis-acting mutations that inactivate a constitutive or alternative splice site within the pre-mRNA, and trans-acting mutations that affect the function of a basal factor of the splicing machinery. Autosomal dominant retinitis pigmentosa (adRP) is caused by mutations in at least 12 genes, with mutations in rhodopsin being the most prevalent. Two cis-acting mutations, g.3811A>G and g.5167G>T at the splice site in the rhodopsin gene (RHO; GenBank U49742.1) are linked to adRP in a Spanish population; while a cis-acting mutation, g.4335G>T, has been linked to recessive RP (arRP). Transcriptional expression analysis showed that the cis-acting splicing mutations linked to adRP promoted alternative splice sites, while the arRP linked mutation results in exclusion of exon 4. Trans-acting splicing mutations associated with adRP have also been found, and mutations in the pre-mRNA splicing factors PRPF3, PRPF8, PRPF31, and RP9 are associated with adRP in several populations. This report describes a new mutation in PRPF3 in a Spanish adRP family. We also investigated the transcriptional patterns in Epstein-Barr virus (EBV)-transformed lymphoblastoid cells from patients carrying a mutation in PRPF8. Despite the role of PRPF8 in the minor U12 splicing processes, microarray analysis revealed that mutations in PRPF8 not only did not result in significant differences in splicing efficiency of rhodopsin, but no apparent changes in expression of U12-type intron genes and splicing processes was observed. Microarray analysis revealed a panel of differentially expressed genes mapped to the RP loci, and future work will determine their role in RP.

Large-scale Molecular Analysis of a 34 Mb Interval on Chromosome 6q: Major Refinement of the RP25 Interval

Annals of Human Genetics. Jul, 2008  |  Pubmed ID: 18510646

A large scale bioinformatics and molecular analysis of a 34 Mb interval on chromosome 6q12 was undertaken as part of our ongoing study to identify the gene responsible for an autosomal recessive retinitis pigmentosa (arRP) locus, RP25. Extensive bioinformatics analysis indicated in excess of 110 genes within the region and we also noted unfinished sequence on chromosome 6q in the Human Genome Database, between 58 and 61.2 Mb. Forty three genes within the RP25 interval were considered as good candidates for mutation screening. Direct sequence analysis of the selected genes in 7 Spanish families with arRP revealed a total of 244 sequence variants, of which 67 were novel but none were pathogenic. This, together with previous reports, excludes 60 genes within the interval ( approximately 55%) as disease causing for RP. To investigate if copy number variation (CNV) exists within RP25, a comparative genomic hybridization (CGH) analysis was performed on a consanguineous family. A clone from the tiling path array, chr6tp-19C7, spanning approximately 100-Kb was found to be deleted in all affected members of the family, leading to a major refinement of the interval. This will eventually have a significant impact on cloning of the RP25 gene.

Linkage Validation of RP25 Using the 10K Genechip Array and Further Refinement of the Locus by New Linked Families

Annals of Human Genetics. Jul, 2008  |  Pubmed ID: 18510647

Retinitis pigmentosa (RP) is a clinically and genetically heterogeneous group of retinal dystrophies, characterised by rod photoreceptor cell degeneration with autosomal recessive RP (arRP) as the commonest form worldwide. To date, a total of 26 loci have been reported for arRP, each having a prevalence of 1-5%, except for the RP25 locus which was identified as the genetic cause of 14% of arRP cases in Spain. In order to validate the original linkage of RP25, we undertook a total genome scan using the 10K GeneChip mapping array on three of the previously linked families. The data obtained supported the initial findings of linkage. Additionally, linkage analysis in 18 newly ascertained arRP families was performed using microsatellite markers spanning the chromosome 6p12.1-q15 interval. Five out of the 18 families showed suggestive evidence of linkage to RP25, hence supporting the high prevalence of this locus in the Spanish population. Furthermore, the finding of a crossover in one of these families is likely to have refined the disease interval from the original 16 cM to only a 2.67 cM region between D6S257 and D6S1557.

Impact of Millennial Mining Activities on Sediments and Microfauna of the Tinto River Estuary (SW Spain)

Marine Pollution Bulletin. Jul, 2008  |  Pubmed ID: 18538354

In this paper, we analyze two short cores collected in the Tinto estuary (SW Spain), and describe the palaeoenvironmental evolution of this area during the last two millennia, along with the influence of historical mining activities and recent industrial pollution on sediments and microfauna (foraminifera and ostracoda). Although there were no significant changes in the distribution of microorganisms, a first pollution period (0-150 AD) was recorded in high sediment pollution by Cu in the shallow palaeochannels of the middle estuary. During this period and the following 1700 years, tolerant pioneer species of both foraminifera and ostracoda were found predominantly in the inner, protected areas of the estuary, while the bottom sediments were subjected to high hydrodynamic gradients, and consequently showed lower density and diversity of organisms. In the last 150 years, acid mine drainage processes, introduction of a new mining period, and the polluted inputs derived from two industrial processes resulted in increased heavy metal contamination of the bottom sediments, and corresponding extirpation of ostracodes and restriction of foraminifers to the inner zones of the estuary.

Fingerprinting the Genetic Diversity of the Biotin Carboxylase Gene (accC) in Aquatic Ecosystems As a Potential Marker for Studies of Carbon Dioxide Assimilation in the Dark

Environmental Microbiology. Oct, 2008  |  Pubmed ID: 18557770

We designed and tested a set of specific primers for specific PCR amplification of the biotin carboxylase subunit gene (accC) of the Acetyl CoA carboxylase (ACCase) enzyme. The primer set yielded a PCR product of c. 460 bp that was suitable for denaturing gradient gel electrophoresis (DGGE) fingerprinting followed by direct sequencing of excised DGGE bands and sequence analysis. Optimization of PCR conditions for selective amplification was carried out with pure cultures of different bacteria and archaea, and laboratory enrichments. Next, fingerprinting comparisons were done in several aerobic and anaerobic freshwater planktonic samples. The DGGE fingerprints showed between 2 and 19 bands in the different samples, and the primer set provided specific amplification in both pure cultures and natural samples. Most of the samples had sequences grouped with bacterial accC, hypothetically related to the anaplerotic fixation of inorganic carbon. Some other samples, however, yielded accC gene sequences that clustered with Crenarchaeota and were related to the 3-hydroxypropionate/4-hydroxybutyrate cycle of autotrophic crenarchaeota. Such samples came from oligotrophic high mountain lakes and the hypolimnia of a sulfide-rich lake, where crenarchaeotal populations had been previously reported by 16S rRNA surveys. This study provided a fast tool to look for presence of accC genes in natural environments as potential marker for studies of carbon dioxide assimilation in the dark. After further refinement for better specificity against archaea, the new and novel primers could be very helpful to establish a target for crenarchaeota with implications for our understanding of archaeal carbon biogeochemistry.

[Videothorascocopic: Approach in Thoracic Inlet of Neurogenic Mediastinal Tumor in Pediatric Age]

Cirugía Pediátrica : Organo Oficial De La Sociedad Española De Cirugía Pediátrica. Apr, 2008  |  Pubmed ID: 18624283

We present a 4 year-old patient with precedent of pneumonia and showing an increase of X-ray density in the superior mediastinal. After a year of follow-up and the presence of irritating cough, it is decided to perform studies of image (CT), being diagnosed of a mass in the region of the thoracic inlet which affects up to D4. Video-assisted thoracic surgery is decided and the mass is entirely removed, being the result of the pathological study to ganglioneuroma. Eight months later from, the patient is asymptomatic and without relapse according to the image methods which were carried out.

NTF-3, a Gene Involved in the Enteric Nervous System Development, As a Candidate Gene for Hirschsprung Disease

Journal of Pediatric Surgery. Jul, 2008  |  Pubmed ID: 18639687

Hirschsprung disease (HSCR) is a congenital disorder caused by a failure of neural crest cells to migrate, proliferate, and/or differentiate during the enteric nervous system (ENS) development. The requirement of the NTF-3/TrkC signaling for the proper development of the ENS, together with the evidences presented by animal models, led us to investigate the involvement of NTF-3 gene in HSCR. We performed both a mutational screening of NTF-3 and a complete evaluation of 3 polymorphisms as genetic susceptibility factors for HSCR. We identified a novel sequence variant, G76R, present in 2 different patients and absent in controls. We postulate that this variation could generate a lack of mature functional NTF-3 proteins in neural crest cell precursors; thus, altering the NTF-3/TrkC signaling pathway and influencing in the adequate ENS development. Although these results do not provide complete assurance of the involvement of this gene in HSCR, given the polygenic nature of the disease and its etiology, investigation of the genes encoding protein members of the signaling pathways governing the ENS development could provide new key findings in the elucidation of this complex disease.

[Disseminated Crusted Papules in a Newborn]

Actas Dermo-sifiliográficas. Sep, 2008  |  Pubmed ID: 18682172

Molecular Typing of Treponema Pallidum Clinical Strains from Lisbon, Portugal

Journal of Clinical Microbiology. Nov, 2008  |  Pubmed ID: 18753355

A molecular system was used to subtype Portuguese Treponema pallidum clinical strains isolated from both skin lesions and blood. The study with this system constitutes the first typing study in a European country. Three T. pallidum subtypes were found: subtypes 14a (50%), 14d (45.2%), and 14f (4.8%). Further studies are needed to better characterize the isolates involved in syphilis outbreaks.

High Archaeal Richness in the Water Column of a Freshwater Sulfurous Karstic Lake Along an Interannual Study

FEMS Microbiology Ecology. Nov, 2008  |  Pubmed ID: 18754782

We surveyed the archaeal assemblage in a stratified sulfurous lake (Lake Vilar, Banyoles, Spain) over 5 consecutive years to detect potential seasonal and interannual trends in the free-living planktonic Archaea composition. The combination of different primer pairs and nested PCR steps revealed an unexpectedly rich archaeal community. Overall, 140 samples were analyzed, yielding 169 different 16S rRNA gene sequences spread over 14 Crenarchaeota (109 sequences) and six Euryarchaeota phylogenetic clusters. Most of the Crenarchaeota (98% of the total crenarchaeotal sequences) affiliated within the Miscellaneous Crenarchaeota Group (MCG) and were related to both marine and freshwater phylotypes. Euryarchaeota mainly grouped within the Deep Hydrothermal Vent Euryarchaeota (DHVE) cluster (80% of the euryarchaeotal sequences) and the remaining 20% distributed into three less abundant taxa, most of them composed of soil and sediment clones. The largest fraction of phylotypes from the two archaeal kingdoms (79% of the Crenarchaeota and 54% of the Euryarchaeota) was retrieved from the anoxic hypolimnion, indicating that these cold and sulfide-rich waters constitute an unexplored source of archaeal richness. The taxon rank-frequency distribution showed two abundant taxa (MCG and DHVE) that persisted in the water column through seasons, plus several rare ones that were only detected occasionally. Differences in richness distribution and seasonality were observed, but no clear correlations were obtained when multivariate statistical analyses were carried out.

The Role of the Humoral Immune Response in the Molecular Evolution of the Envelope C2, V3 and C3 Regions in Chronically HIV-2 Infected Patients

Retrovirology. 2008  |  Pubmed ID: 18778482

This study was designed to investigate, for the first time, the short-term molecular evolution of the HIV-2 C2, V3 and C3 envelope regions and its association with the immune response. Clonal sequences of the env C2V3C3 region were obtained from a cohort of eighteen HIV-2 chronically infected patients followed prospectively during 2-4 years. Genetic diversity, divergence, positive selection and glycosylation in the C2V3C3 region were analysed as a function of the number of CD4+ T cells and the anti-C2V3C3 IgG and IgA antibody reactivity

C-Lysozyme from Senegalese Sole (Solea Senegalensis): CDNA Cloning and Expression Pattern

Fish & Shellfish Immunology. Nov, 2008  |  Pubmed ID: 18786641

Lysozymes are key molecules of innate immunity and proved high bactericidal activity in fish, thus becoming attractive as tools for enhancing fish defences. In this study, a full-length c-type lysozyme cDNA from Senegalese sole (Solea senegalensis) has been cloned and characterized. The cDNA sequence was inferred from two overlapping fragments obtained by RACE-PCR and consisting on 631bp coding for 143 aminoacids. Catalytic and other conserved residues required for lysozyme activity were identified. Pair wise alignments showed the higher identities with c-type lysozyme from other flatfish. Expression patterns under various conditions showed a basal level and a clear upregulation mostly in hematopoietic organs after stimulation with LPS or infection with Photobacterium damselae. This study represents a first step on the genetics and function of the c-lysozyme of Senegalese sole, though disclosing g-DNA structure, allelic variability and antibacterial activity must be requirements prior its immunological properties might have biotechnological applications.

In Vitro Inhibition of Sole Aquabirnavirus by Senegalese Sole Mx

Fish & Shellfish Immunology. Feb, 2008  |  Pubmed ID: 18078763

Senegalese sole, Solea senegalensis, is a flat fish of growing interest in European aquaculture. In its culture viral infections are constant threats, thus understanding antiviral defences is a key factor for a successful industry. Mx proteins are IFN-induced proteins widespread in eukaryotes; however, their antiviral activity is unclear and the results variable among species. Therefore assessment of the putative Mx antiviral activity in each species is of interest. Our group has recently cloned the Senegalese sole Mx (SsMx) cDNA and in this study its antiviral activity was assessed by infecting CHSE-214 cells expressing recombinant SsMx, with sole aquabirnavirus. The antiviral activity against this pathogen was demonstrated by reduction in induced cytopathic effects, reduction in virus yield and decrease in viral transcripts. These findings contribute to our understanding of fish antiviral mechanisms and open the possibility of using this protein as a tool for fighting viral infections in aquaculture.

Genetic Analysis of FAM46A in Spanish Families with Autosomal Recessive Retinitis Pigmentosa: Characterisation of Novel VNTRs

Annals of Human Genetics. Jan, 2008  |  Pubmed ID: 17803723

Retinitis pigmentosa (RP) is a group of retinal dystrophies characterised primarily by rod photoreceptor cell degeneration. Exhibiting great clinical and genetic heterogeneity, RP be inherited as an autosomal dominant (ad) and recessive (ar), X-linked (xl) and digenic disorder. RP25, a locus for arRP, was mapped to chromosome 6p12.1-q14.1 where several retinal dystrophy loci are located. A gene expressed in the retina, FAM46A, mapped within the RP25 locus, and computational data revealed its involvement in retinal signalling pathways. Therefore, we chose to perform molecular evaluation of this gene as a good candidate in arRP families linked to the RP25 interval. A comprehensive bioinformatic and retinal tissue expression characterisation of FAM46A was performed, together with mutation screening of seven RP25 families. Herein we present 4 novel sequence variants, of which one is a novel deletion within a low complexity region close to the initiation codon of FAM46A. Furthermore, we have characterised for the first time a coding tandem variation in the Caucasian population. This study reports on bioinformatic and moleculardata for the FAM46A gene that may give a wider insight into the putative function of this gene and its pathologic relevance to RP25 and other retinal diseases mapping within the 6q chromosomal interval.

Recent Advances in the Development of Recombinant Vaccines Against Classical Swine Fever Virus: Cellular Responses Also Play a Role in Protection

Veterinary Journal (London, England : 1997). Aug, 2008  |  Pubmed ID: 17804267

Classical swine fever virus (CSFV) is the causative agent of one of the most devastating porcine haemorrhagic viral diseases, classical swine fever (CSF). CSFV mainly infects endothelial cells and macrophages and at the same time promotes bystander apoptosis of the surrounding T cells, causing strong immune suppression and high mortality rates. Most animals experience acute infection, during which they either die or survive by producing neutralising antibodies to the virus. However, in a few cases, the impaired immune system cannot control viral progression, leading to chronic infection. Efficient live attenuated vaccines against CSFV exist and are routinely used only in endemic countries. The ability of these vaccines to replicate in the host, even at very low rates, makes it extremely difficult to distinguish vaccinated from infected animals, favouring a restricted policy regarding vaccination against CSFV in non-endemic countries. There is a clear need for efficient and safer marker vaccines to assist in the control of future CSF outbreaks. In this review article, some of the most recent advances in the field of recombinant vaccines against CSFV are presented and the nature of the protective immune responses they induce is discussed.

Hirschsprung Disease, Associated Syndromes and Genetics: a Review

Journal of Medical Genetics. Jan, 2008  |  Pubmed ID: 17965226

Hirschsprung disease (HSCR, aganglionic megacolon) represents the main genetic cause of functional intestinal obstruction with an incidence of 1/5000 live births. This developmental disorder is a neurocristopathy and is characterised by the absence of the enteric ganglia along a variable length of the intestine. In the last decades, the development of surgical approaches has importantly decreased mortality and morbidity which allowed the emergence of familial cases. Isolated HSCR appears to be a non-Mendelian malformation with low, sex-dependent penetrance, and variable expression according to the length of the aganglionic segment. While all Mendelian modes of inheritance have been described in syndromic HSCR, isolated HSCR stands as a model for genetic disorders with complex patterns of inheritance. The tyrosine kinase receptor RET is the major gene with both rare coding sequence mutations and/or a frequent variant located in an enhancer element predisposing to the disease. Hitherto, 10 genes and five loci have been found to be involved in HSCR development.

Expression Analysis of Mx Protein and Evaluation of Its Antiviral Activity Against Sole Aquabirnavirus in SAF-1 and TV-1 Cell Lines

Veterinary Immunology and Immunopathology. Jan, 2008  |  Pubmed ID: 17981342

The transcription of Mx mRNA after poly I:C induction and sole aquabirnavirus infection has been analysed in SAF-1 and TV-1 cells (derived from gilt-head seabream and turbot, respectively). Both cell lines were stimulated with 10 microg ml(-1) poly I:C and Mx mRNA was analysed by a specific RT-PCR at several times post-induction. The results showed a high level of Mx expression from 12 to 120 h after induction in SAF-1 cells, whereas in TV-1 cells Mx mRNA was only detected at 12 and 24h. The treatment with different concentrations of poly I:C showed that TV-1 cells are less sensitive to this inductor than the SAF-1 cell line. The antiviral activity derived from poly I:C induction has been clearly demonstrated against sole aquabirnavirus on both cell lines. The inoculation of sole aquabirnavirus resulted in the Mx mRNA transcription at 48, 72, and 96 h post-infection (p.i.) in SAF-1 cells. On the contrary, inoculated TV-1 cells only showed a faint Mx mRNA band at 24 and 48 h p.i. This study has established different patterns of Mx expression in both cells under study as a consequence of the poly I:C induction and sole aquabirnavirus infection, and it shows that gilt-head seabream and turbot Mx inhibit sole aquabirnavirus replication.

[Use of Transparencies to Read Patch Tests]

Actas Dermo-sifiliográficas. Nov, 2008  |  Pubmed ID: 19087818

Study of the Effect of Sample Preparation and Cooking on the Selenium Speciation of Selenized Potatoes by HPLC with ICP-MS and Electrospray Ionization MS/MS

Journal of Agricultural and Food Chemistry. Jan, 2009  |  Pubmed ID: 19093878

The efficiency of enzymatic hydrolysis and leaching with water using accelerated solvent extraction (ASE) or boiling was investigated for quantitative Se speciation in selenized potatoes using reversed phase HPLC coupled to ICP-MS. Preliminary identification of selenomethionine (SeMet), Se-methylselenocysteine (SeMeCys), and selenate in extracts of potato skin and flesh was achieved using complementary reversed phase and anion-exchange HPLC-ICP-MS and retention time matching with standards. The quantitative speciation data revealed a higher percentage of selenomethionine (73% of the total Se) found in the flesh in comparison with skin (containing 21% of the total Se as SeMet). ASE and boiling in water were found to be similar in terms of Se extraction efficiency and profiles. However, ASE was found to be more efficient than boiling with respect to sample cleanup and reduced sample handling. The presence of SeMet at parts per billion levels in selenized potatoes was confirmed by reversed phase HPLC with online ESI MS/MS.

Lymphogranuloma Venereum in Portugal: Unusual Events and New Variants During 2007

Sexually Transmitted Diseases. Feb, 2009  |  Pubmed ID: 19125144

Several European countries identified an ongoing LGV outbreak, particularly among men who have sex with men (MSM). In Portugal, no particular surveillance measures were launched. Nonetheless, circulating LGV strains could eventually be detected through the routine Chlamydia trachomatis ompA genotyping procedure held in the Portuguese National Institute of Health (NIH).

Attenuated Foot-and-mouth Disease Virus RNA Carrying a Deletion in the 3' Noncoding Region Can Elicit Immunity in Swine

Journal of Virology. Apr, 2009  |  Pubmed ID: 19211755

We constructed foot-and-mouth disease virus (FMDV) mutants bearing independent deletions of the two stem-loop structures predicted in the 3' noncoding region of viral RNA, SL1 and SL2, respectively. Deletion of SL2 was lethal for viral infectivity in cultured cells, while deletion of SL1 resulted in viruses with slower growth kinetics and downregulated replication associated with impaired negative-strand RNA synthesis. With the aim of exploring the potential of an RNA-based vaccine against foot-and-mouth disease using attenuated viral genomes, full-length chimeric O1K/C-S8 RNAs were first inoculated into pigs. Our results show that FMDV viral transcripts could generate infectious virus and induce disease in swine. In contrast, RNAs carrying the DeltaSL1 mutation on an FMDV O1K genome were innocuous for pigs but elicited a specific immune response including both humoral and cellular responses. A single inoculation with 500 microg of RNA was able to induce a neutralizing antibody response. This response could be further boosted by a second RNA injection. The presence of the DeltaSL1 mutation was confirmed in viruses isolated from serum samples of RNA-inoculated pigs or after transfection and five passages in cell culture. These findings suggest that deletion of SL1 might contribute to FMDV attenuation in swine and support the potential of RNA technology for the design of new FMDV vaccines.

[Lichenoid Eruption in a Patient with Chronic Hepatitis C Virus Infection Treated with Interferon and Ribavirin]

Gastroenterología Y Hepatología. Feb, 2009  |  Pubmed ID: 19231693

Cytarabine-associated Lymphoid Dyscrasia with Atypical T CD30+ Infiltrate in a Patient Suffering from Acute Nonlymphoblastic Leukaemia

The British Journal of Dermatology. May, 2009  |  Pubmed ID: 19292713

Interaction Between a Chromosome 10 RET Enhancer and Chromosome 21 in the Down Syndrome-Hirschsprung Disease Association

Human Mutation. May, 2009  |  Pubmed ID: 19306335

Individuals with Down syndrome (DS) display a 40-fold greater risk of Hirschsprung disease (HSCR) than the general population of newborns implicating chromosome 21 in HSCR etiology. Here we demonstrate that the RET enhancer polymorphism RET+9.7 (rs2435357:C>T) at chromosome 10q11.2 is associated with HSCR in DS individuals both by transmission disequilibrium (P=0.0015) and case-control (P=0.0115) analysis of matched cases. Interestingly, the RET+9.7 T allele frequency is significantly different between individuals with DS alone (0.26+/-0.04), HSCR alone (0.61+/-0.04), and those with HSCR and DS (0.41+/-0.04), demonstrating an association and interaction between RET and chromosome 21 gene dosage. This is the first report of a genetic interaction between a common functional variant (rs2435357) and a not infrequent copy number error (chromosome 21 dosage) in two human developmental disorders.

Sleep-disordered Breathing and Acute Stroke

Cerebrovascular Diseases (Basel, Switzerland). 2009  |  Pubmed ID: 19342839

Sleep-disordered breathing (SDB) is a disease of increasing importance and it is frequent in stroke patients. SDB is being recognized as an independent risk factor for several clinical consequences, including cardiovascular and cerebrovascular disease.

A Single Residue, Arginine 65, is Critical for the Functional Interaction of Leukocyte-associated Inhibitory Receptor-1 with Collagens

Journal of Immunology (Baltimore, Md. : 1950). May, 2009  |  Pubmed ID: 19380792

ITIM-containing receptors play an essential role in modulating immune responses. Leukocyte-associated inhibitory receptor (LAIR)-1, also known as CD305, is an ITIM-containing inhibitory receptor, expressed by all leukocytes, that binds collagens. In this article, we investigate the effect of a conservative R65K mutation on LAIR-1 ligand binding and function. Compared with LAIR-1 wild-type (wt)-expressing cells, LAIR-1 R65K cells show markedly reduced binding to collagen, which correlates with a reduced level of LAIR-1 polarization to the site of interaction with collagens. Both LAIR-1 wt and R65K cells can generate intracellular signals when ligated by anti-LAIR-1 mAb, but only LAIR-1 wt cells respond to collagens or matrigel. In agreement, surface plasmon resonance analyses showed that LAIR-1 R65K protein has markedly reduced avidity for collagen type I compared with LAIR-1 wt. Likewise, LAIR-1 R65K protein has decreased avidity for cells expressing transmembrane collagen XVII. Thus, a single residue, Arg65, is critical for the interaction of LAIR-1 with collagens.

Treatment of Feline Mammary Tumours Using Chemotherapy, Surgery and a COX-2 Inhibitor Drug (meloxicam): a Retrospective Study of 23 Cases (2002-2007)*

Veterinary and Comparative Oncology. Dec, 2009  |  Pubmed ID: 19891691

The efficacy of a treatment combination of a COX-2 inhibitor (meloxicam), chemotherapy and surgery in 23 cats with histologically confirmed mammary gland adenocarcinoma was evaluated. All of the cases underwent an aggressive surgery with concurrent doxorubicin-based chemotherapy. Meloxicam was given orally starting the day after surgery and was continued indefinitely. Serum renal parameters were measured every 3-5 months. Three cats developed azotemia, whereas in four other renal parameters increased but remained within normal limit. The Kaplan-Meier median survival time was 460 days. The Kaplan-Meier median disease free interval was 269 days. The survival times are similar to other studies, not supporting the use of this treatment combination. Prospective studies with a higher number of cases are warranted to investigate the utility of this multimodality protocol for the treatment of feline mammary tumours.

Leflunomide in the Treatment of Palmoplantar Pustulosis

Archives of Dermatology. Nov, 2009  |  Pubmed ID: 19917950

A Novel Study of Copy Number Variations in Hirschsprung Disease Using the Multiple Ligation-dependent Probe Amplification (MLPA) Technique

BMC Medical Genetics. 2009  |  Pubmed ID: 19925665

Hirschsprung disease (HSCR) is a congenital malformation of the hindgut produced by a disruption in neural crest cell migration during embryonic development. HSCR has a complex genetic etiology and mutations in several genes, mainly the RET proto-oncogene, have been related to the disease. There is a clear predominance of missense/nonsense mutations in these genes whereas copy number variations (CNVs) have been seldom described, probably due to the limitations of conventional techniques usually employed for mutational analysis.

Lymphogranuloma Venereum in Europe, 2003-2008

Euro Surveillance : Bulletin Européen Sur Les Maladies Transmissibles = European Communicable Disease Bulletin. 2009  |  Pubmed ID: 20003898

Lymphogranuloma venereum, caused by the L serovars of Chlamydia trachomatis, emerged in Europe in 2003 and a series of outbreaks were reported in different countries. The infection presents as a severe proctitis in men who have sex with men, many of whom are co-infected with HIV and other sexually transmitted infections. This paper reviews the number of cases reported over a five year period, from 2003 to 2008, from countries that were part of the European Surveillance of Sexually Transmitted Infections (ESSTI) network. Reports were received from Belgium, Denmark, France, Germany, the Netherlands, Portugal, Spain, Sweden, and the United Kingdom. It appears that after five years the characteristics of the patients infected has overall remained unchanged, although the total number of cases has increased and more countries in Europe have now identified cases of LGV.

Temporary Henna Tattoos with Long-term Consequences

The Medical Journal of Australia. Dec 7-21, 2009  |  Pubmed ID: 20028309

Regulatory Cells, Cytokine Pattern and Clinical Risk Factors for Asthma in Infants and Young Children with Recurrent Wheeze

Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology. Aug, 2009  |  Pubmed ID: 19438590

Several risk factors for asthma have been identified in infants and young children with recurrent wheeze. However, published literature has reported contradictory findings regarding the underlying immunological mechanisms.

Availability of Glucose and Light Modulates the Structure and Function of a Microbial Biofilm

FEMS Microbiology Ecology. Jul, 2009  |  Pubmed ID: 19453745

We have studied the differences in the organic matter processing and biofilm composition and structure between autoheterotrophic and heterotrophic biofilm communities. Microbial communities grown on artificial biofilms were monitored, following incubation under light and dark conditions and with or without the addition of glucose as a labile organic compound. Glucose addition greatly affected the microbial biofilm composition as shown by differences in 16S rRNA gene fingerprints. A significant increase in beta-glucosidase and peptidase enzyme activities were also observed in glucose-amended biofilms incubated in the dark, suggesting an active bacterial community. Light enhanced the algal and bacterial growth, as well as higher extracellular enzyme activity, thereby indicating a tight algal-bacterial coupling in biofilms incubated under illumination. In these biofilms, organic compounds excreted by photosynthetic microorganisms were readily available for bacterial heterotrophs. This algal-bacterial relationship weakened in glucose-amended biofilms grown in the light, probably because heterotrophic bacteria preferentially use external labile compounds. These results suggest that the availability of labile organic matter in the flowing water and the presence of light may alter the biofilm composition and function, therefore affecting the processing capacity of organic matter in the stream ecosystem.

[Update of the Topical Treatment of Psoriasis]

Actas Dermo-sifiliográficas. Apr, 2009  |  Pubmed ID: 19457304

Topical therapy continues to be one of the pillars of psoriasis management. Topical corticosteroids and vitamin D analogs are the drugs of choice during the induction phase, and vitamin D analogs continue to be drugs of choice for maintenance therapy. Tazarotene and dithranol are suitable options in patients with certain, specific characteristics. The calcineurin inhibitors can be considered to be second-line treatment for psoriasis of the face and flexures. The efficacy and safety of the fixed-dose combination of betamethasone and calcipotriol in the induction phase is greater than that of either drug alone. The combination of corticosteroids with salicylic acid achieves better results than corticosteroids in monotherapy. None of the drugs evaluated stands out over the others in all clinical situations, and their use must therefore be individualized in each patient and adjusted according to the course of the disease.

Calcinosis Cutis Associated with Amyopathic Dermatomyositis: Response to Intravenous Immunoglobulin

Journal of the American Academy of Dermatology. Jun, 2009  |  Pubmed ID: 19467386

Histopathologic Characteristics of Neonatal Cutaneous Lupus Erythematosus: Description of Five Cases and Literature Review

Journal of Cutaneous Pathology. Jun, 2009  |  Pubmed ID: 19515045

Neonatal lupus erythematosus (NLE) is a disease associated with the transplacental transfer of maternal anti-Ro/SSA. The histopathologic characteristics of neonatal lupus have been described as compatible with cutaneous lupus based on isolated cases.

Ozone Oxidative Preconditioning Reduces Nitrite Levels in Blood Serum in LPS: Induced Endotoxic Shock in Mice

Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]. Aug, 2009  |  Pubmed ID: 19533019

Reactive oxygen species, and also reactive species of nitrogen such as nitric oxide, play an important role in the pathogenesis of peritonitis and septic shock. Ozone oxidative preconditioning (OOP) has shown protective effects in various experimental models of peritonitis in rats and endotoxic shock in mice. Currently, strong evidence is available that this protective effect of OOP is due to its action on the balance between endogenous antioxidants and pro-oxidants, which is favorable for anti-oxidant defense. The aim of this research was to elucidate whether or not OOP is able to reduce nitrite levels in blood serum of mice treated with lipopolysaccharide (LPS). We used an experimental model of endotoxic shock induced by LPS in mice in which the animals were pre-treated with ozone/oxygen mixture for 5 days (once daily), with injection of LPS 24 h thereafter to induce endotoxic shock.

Trends in Kidney Transplantation Outcome: the Andalusian Kidney Transplant Registry, 1984-2007

Transplantation Proceedings. Jun, 2009  |  Pubmed ID: 19545684

Herein we have presented the first report from the Andalusian Kidney Transplant Registry, a Public Health Service Regional Registry in Andalusia, Spain (general population, 8 million). The current analysis was limited to 5599 kidney-alone transplants from deceased donors, grouped into 4 time periods: 1984-1989 (n = 846); 1990-1995 (n = 1172); 1996-2001 (n = 1801); and 2002-2007 (n = 2060). The age of the transplant patients rose over time to 21.7% of recipients of ages >or=60 years in 2002-2007. In the later years we observed an increased incidence of vascular and diabetic causes of end-stage renal disease (ESRD). Patients who underwent retransplantation increased from 2.7% in 1984-1989 to 8.1% in 2002-2007. Time on previous renal replacement therapy (RRT) increased from 33.1 +/- 29 to 48 +/- 53 months. Patient survivals at 1, 5, 10, and 20 years were 96%, 91%, 83%, and 63%, respectively. Censoring for death, graft survivals were 90%, 80%, 67%, and 45%, respectively. Compared with the 1984-1989 period, patient survival improved by about 10% (P < .001) since 1990, remaining stable to 2007. Censored 5-year graft survivals progressively improved from 72% to 77%, 82%, and 85% (P < .001). Upon multivariate analysis, gender, age >39 years, diabetes, and RRT duration were independent predictors of patient survival. Age <18 years, retransplantation, and positive hepatitis C virus serology were independent predictors of lower graft survival. Considering 1984-1989 as the reference time period, both patient and graft mortality risks continuously decreased over the following 3 periods (relative risk [RR] = 0.5-0.4-0.3 for patient mortality; RR = 0.8-0.6-0.5 for graft mortality). In summary, despite an increased number of adverse risk factors, both patient and graft survivals have improved from 1984 to date.

Contribution of RET, NTRK3 and EDN3 to the Expression of Hirschsprung Disease in a Multiplex Family

Journal of Medical Genetics. Dec, 2009  |  Pubmed ID: 19556619

BACKGROUND: Hirschsprung disease (HSCR) is a developmental disorder caused by a defect in the neural crest neuroblast migration process. It is considered to be a paradigm of complex disorders, with many loci contributing to manifestation of the disease. Although HSCR commonly appears as a sporadic trait, approximately 20% of HSCR cases are familial, with complex patterns of inheritance. METHOD: A multiplex HSCR family with an additive model of inheritance, in which the contribution of three genes (RET, NTRK3, EDN3) leads to the HSCR phenotype is reported. Results and discussion: The findings suggest that both RET and NTRK3 mutations acting together are necessary and sufficient for the appearance of the disease, and that the EDN3 mutation is acting as a phenotype-modifier factor in the context of this family, as two different HSCR phenotypes are seen among the affected members: a short segment form, and a total colonic aganglionosis. The results therefore support the complex additive model of inheritance previously proposed for Hirschsprung disease.

Predation Impact of Ciliated and Flagellated Protozoa During a Summer Bloom of Brown Sulfur Bacteria in a Meromictic Coastal Lake

FEMS Microbiology Ecology. Oct, 2009  |  Pubmed ID: 19622068

Anaerobic phagotrophic protozoa may play an important role in the carbon flux of chemically stratified environments, especially when phototrophic sulfur bacteria account for a high proportion of the primary production. To test this assumption, we investigated the vertical and temporal distribution of microbial heterotrophs and of autotrophic picoplankton throughout the water column of the meromictic coastal lake Faro (Sicily, Italy), in the summer of 2004, coinciding with a bloom of brown-colored green sulfur bacteria. We also assessed the grazing impact of ciliated and flagellated protozoa within the sulfur bacteria plate using a modification of the fluorescently labeled bacteria uptake approach, attempting to minimize the biases intrinsic to the technique and to preserve the in situ anoxic conditions. Significant correlations were observed between ciliate biomass and bacteriochlorophyll e concentration, and between heterotrophic nanoflagellate biomass and chlorophyll a concentration in the water column. The major predators of anaerobic picoplankton were pleuronematine ciliates and cryptomonad flagellates, with clearances of 26.6 and 9.5 nL per cell h(-1), respectively, and a cumulative impact on the picoplankton gross growth rate ranging between 36% and 72%. We concluded that protozoan grazing channels a large proportion of anaerobic picoplankton production to higher trophic levels without restraining photosynthetic bacteria productivity.

Dermatologists in Hospital Wards: an 8-year Study of Dermatology Consultations

Dermatology (Basel, Switzerland). 2009  |  Pubmed ID: 19648729

The value of dermatologists as consultants is increasing.

The RET Functional Variant C 587T>C is Not Associated with Susceptibility to Sporadic Medullary Thyroid Cancer

Thyroid : Official Journal of the American Thyroid Association. Sep, 2009  |  Pubmed ID: 19678735

Molecular Markers for Establishing Distinctness in Vegetatively Propagated Crops: a Case Study in Grapevine

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik. Nov, 2009  |  Pubmed ID: 19680623

Distinctness, uniformity and stability (DUS) testing of varieties is usually required to apply for Plant Breeders' Rights. This exam is currently carried out using morphological traits, where the establishment of distinctness through a minimum distance is the key issue. In this study, the possibility of using microsatellite markers for establishing the minimum distance in a vegetatively propagated crop (grapevine) has been evaluated. A collection of 991 accessions have been studied with nine microsatellite markers and pair-wise compared, and the highest intra-variety distance and the lowest inter-variety distance determined. The collection included 489 different genotypes, and synonyms and sports. Average values for number of alleles per locus (19), Polymorphic Information Content (0.764) and heterozygosities observed (0.773) and expected (0.785) indicated the high level of polymorphism existing in grapevine. The maximum intra-variety variability found was one allele between two accessions of the same variety, of a total of 3,171 pair-wise comparisons. The minimum inter-variety variability found was two alleles between two pairs of varieties, of a total of 119,316 pair-wise comparisons. In base to these results, the minimum distance required to set distinctness in grapevine with the nine microsatellite markers used could be established in two alleles. General rules for the use of the system as a support for establishing distinctness in vegetatively propagated crops are discussed.

Are We Still Making Progress in Patient Survival After Kidney Transplantation? Results of a Regional Registry

Transplantation Proceedings. Jul-Aug, 2009  |  Pubmed ID: 19715838

Many studies have shown a trend to improved long-term survival of renal transplant recipients. We analyzed the survival of recipients in Andalusia, Spain, from 1984-2007. The study included all the deceased donor, non-multiorgan grafts (n = 5599), grouped over successive 6-year periods, compared for corrected recipient survival. Changes were noted in the recipient characteristics: increased age, diabetes, vascular nephropathy, retransplantation, duration of prior replacement therapy, and reduction in positive hepatitis C virus (HCV+) serology. The univariate analysis showed a significantly worse survival associated with increased age (P < .001), diabetes (P < .001), HCV+ serology (P < .01; 1996-2007), and longer times on replacement therapy, but not with sex or retransplantation. The respective survivals at 1, 5, and 10 years in 1984-1989 were 93%, 86%, and 75%; in 1990-1995, 97%, 92%, and 84%; in 1996-2001, 96%, 91%, and 84%; and in 2002-2007, 96% and 92%, respectively. There was a significant improvement between the first and second periods (P < .001), but no change thereafter. The multivariate analysis (Cox) showed, a significant influence of age >40 years, female gender (relative risk [RR] 0.8; 95% confidence interval [CI] 0.7-0.9), diabetes (RR 2.5; 95% CI 1.8-3.4), and duration of prior replacement therapy (RR 1.08; 95% CI 1.05-1.1). The risk varied significantly depending on the period: using 2002-2007 as the reference period, the RR in 1984-1989 was 3.4 (95% CI 2.6-4.5); in 1990-1995, 1.8 (95% CI 1.3-2.3); and in 1996-2001, 1.4 (95% CI 1.1-1.8; all P < .02). The model remained for 1996-2007, though HCV+ serology was not significant. In conclusion, we showed a significant improvement in recipient survival in Andalusia over time. Correction for worse recipient characteristics suggests continued advances.

Endocytosis and Intracellular Trafficking of Human Natural Killer Cell Receptors

Traffic (Copenhagen, Denmark). Dec, 2009  |  Pubmed ID: 19719476

Natural killer (NK) cells play a vital role in the defense against viral infections and tumor development. NK cell function is primarily regulated by the sum of signals from a broad array of activation and inhibitory receptors. Key to generating the input level of either activating or inhibitory signals is the maintenance of receptor expression levels on the cell surface. Although the mechanisms of endocytosis and trafficking for some cell surface receptors, such as transferrin receptor and certain immune receptors, are very well known, that is not the situation for receptors expressed by NK cells. Recent studies have uncovered that endocytosis and trafficking routes characteristic for specific activation and inhibitory receptors can regulate the functional responses of NK cells. In this review, we summarize the current knowledge of receptor endocytosis and trafficking, and integrate this with our current understanding of NK cell receptor trafficking.

Gene Expression Profiles of Bone Marrow Cells from Mice Phenotype-selected for Maximal or Minimal Acute Inflammations: Searching for Genes in Acute Inflammation Modifier Loci

Immunology. Sep, 2009  |  Pubmed ID: 19740317

Two mouse lines were phenotype-selected for maximum (AIRmax) or minimum (AIRmin) acute inflammation responses to polyacrylamide bead (Biogel) injection. These lines differ in terms of bone marrow granulopoiesis, neutrophil resistance to apoptosis, and inflammatory cytokine production during acute inflammation responses. We compared gene expression profiles in bone marrow cells (BMC) of AIRmax and AIRmin mice during acute inflammatory reactions. The BMC from femurs were recovered 24 hr after subcutaneous injections of Biogel. Global gene expression analysis was performed on CodeLink Bioarrays (36K genes) using RNA pools of BMC from both control and treated AIRmax and AIRmin mice. Differentially expressed genes were statistically established and the over-represented gene ontology biological process categories were identified. Upregulations of about 136 and 198 genes were observed in the BMC of Biogel-treated AIRmax and AIRmin mice, respectively, but 740 genes were found to be downregulated in AIRmin mice compared with 94 genes in AIRmax mice. The over-represented biological themes of the differently expressed genes among AIRmax and AIRmin mice represent inflammatory response, signal transduction, cell proliferation and immune cell chemotaxis. We were able to demonstrate a broad downmodulation of gene transcripts in BMC from AIRmin mice during acute inflammation, and significant differentially expressed genes colocalized with previously mapped regions for inflammation-related phenotypes in chromosomes 1, 3, 6 and 11.

Allergy to Extensively Hydrolysed Formulas

Allergologia Et Immunopathologia. Sep-Oct, 2009  |  Pubmed ID: 19769840

Skin Prick Tests and Allergy Diagnosis

Allergologia Et Immunopathologia. May-Jun, 2009  |  Pubmed ID: 19769849

Skin testing remains an essential diagnostic tool in modern allergy practice. A significant variability has been reported regarding technical procedures, interpretation of results and documentation. This review has the aim of consolidating methodological recommendations through a critical analysis on past and recent data. This will allow a better understanding on skin prick test (SPT) history; technique; (contra-) indications; interpretation of results; diagnostic pitfalls; adverse reactions; and variability factors.

[Refractory Subacute Cutaneous Lupus Erythematosus with a Response to Efalizumab]

Actas Dermo-sifiliográficas. Oct, 2009  |  Pubmed ID: 19775560

Evolutionary Dynamics of OmpA, the Gene Encoding the Chlamydia Trachomatis Key Antigen

Journal of Bacteriology. Dec, 2009  |  Pubmed ID: 19783629

Chlamydia trachomatis is the trachoma agent and causes most bacterial sexually transmitted infections worldwide. Its major outer membrane protein (MOMP) is a well-known porin and adhesin and is the dominant antigen. So far, investigation of MOMP variability has been focused mainly on molecular epidemiological surveys. In contrast, we aimed to evaluate the impact of the host pressure on this key antigen by analyzing its evolutionary dynamics in 795 isolates from urogenital infections, taking into account the MOMP secondary structure and the sizes/positions of antigenic regions. One-third of the specimens showed a mutational drift from the corresponding genotype, where approximately 42% of the mutations had never been described. Amino acid alterations were sixfold more frequent within B-cell epitopes than in the remaining protein (P = 0.027), and some mutations were also found within or close to T-cell antigenic clusters. Interestingly, the two most ecologically successful genotypes, E and F, showed a mutation rate 60.3-fold lower than that of the other genotypes (P < 10(-8)), suggesting that their efficacy may be the result of a better fitness in dealing with the host immune system rather than of specific virulence factors. Furthermore, the variability exhibited by some genetic variants involved residues that are known to play a critical role during the membrane mechanical movements, contributing to a more stable and flexible porin conformation, which suggests some plasticity to deal with environmental pressure. Globally, these MOMP mutational trends yielded no mosaic structures or important phylogenetic changes, but instead yielded point mutations on specific protein domains, which may enhance pathogen's infectivity, persistence, and transmission.

Contact Urticaria Due to Phenoxyethanol in an Aftershave

Dermatitis : Contact, Atopic, Occupational, Drug : Official Journal of the American Contact Dermatitis Society, North American Contact Dermatitis Group. Aug, 2009  |  Pubmed ID: 19804693

The occurrence of pruritus immediately after application of an aftershave product is usually due to irritant contact dermatitis. We report a case of contact urticaria in a male patient, produced by an aftershave product containing phenoxyethanol.

Application of in Situ Detection Techniques to Determine the Systemic Condition of Lymphocystis Disease Virus Infection in Cultured Gilt-head Seabream, Sparus Aurata L

Journal of Fish Diseases. Feb, 2009  |  Pubmed ID: 18803582

Immunohistochemistry (IHC) and in situ hybridization (ISH) techniques have been used for the detection of lymphocystis disease virus (LCDV) in formalin-fixed, paraffin-embedded tissues from gilt-head seabream, Sparus aurata L. Diseased and recovered fish from the same population were analysed. IHC was performed with a polyclonal antibody against a 60-kDa viral protein. A specific digoxigenin-labelled probe, obtained by PCR amplification of a 270-bp fragment of the gene coding the LCDV major capsid protein, was used for ISH. LCDV was detected in skin dermis and gill lamellae, as well as in several internal organs such as the intestine, liver, spleen and kidney using both techniques. Fibroblasts, hepatocytes and macrophages seem to be target cells for virus replication. The presence of lymphocystis cells in the dermis of the skin and caudal fin, and necrotic changes in the epithelium of proximal renal tubules were the only histological alterations observed in fish showing signs of the disease.

Quantification of the Effect of Nonphotochemical Quenching on the Determination of in Vivo Chl a from Phytoplankton Along the Water Column of a Freshwater Reservoir

Photochemistry and Photobiology. Jan-Feb, 2009  |  Pubmed ID: 18811624

Nonphotochemical quenching (NPQ) is a well-known collection of different photoprotective mechanisms of plants and algae to avoid photodamage under an excess of light energy. In order to evaluate the overall effect of NPQ processes on the fluorometric determination of in vivo Chl a from a phytoplankton community dominated by diatoms, we compared the results obtained by two different fluorometric field devices with the total concentration of extracted Chl a measured by HPLC (in vitro Chl a). A different set of measurements were made to assess the performance of these fluorometers at high, moderate and low irradiance conditions. The Fbbe fluorometer, which is capable of distinguishing different algal groups according to their pigment content, allowed a better determination of in vivo Chl a under high irradiance conditions, with only a 10% mean difference from the in vitro Chl a concentration. In turn, the FMII fluorometer underestimated by as much as 50% the in vitro Chl a concentration under the same light conditions. As data from both fluorometers were in accordance with the in vitro Chl a values at moderate irradiance levels, the differences observed at high irradiances were attributed to the decrease in the yield of Chl a fluorescence caused by photoprotective NPQ processes. Accordingly, we estimated the effect of NPQ processes on the in vivo Chl a determination and the results allow us to provide an equation to correct this effect when in situ fluorometric measurements are carried out under high irradiance regimes. Our results demonstrate that under certain circumstances NPQ seriously compromises the results obtained by in situ fluorometric probes and highlight the need for a cautious interpretation of field data under such environmental conditions.

A Novel Point Variant in NTRK3, R645C, Suggests a Role of This Gene in the Pathogenesis of Hirschsprung Disease

Annals of Human Genetics. Jan, 2009  |  Pubmed ID: 19040714

Hirschsprung disease (HSCR) is a developmental disorder characterized by the absence of ganglion cells in the myenteric and submucosal plexuses due to a defect in the migration process of neural crest neuroblasts. Manifestation of the disease has been linked to the dysfunction of two principal signalling pathways involved in the enteric nervous system (ENS) formation: the RET-GDNF and the EDN3-EDNRB receptor systems. However, the NTF3/NTRK3 signalling pathway plays an essential role in the development of the ENS suggesting a potential role for those genes in the pathogenesis of HSCR. We have sought to evaluate the candidature of the NTRK3 gene, which encodes the TrkC receptor, as a susceptibility gene for Hirschsprung disease. Using dHPLC technology we have screened the NTRK3 coding region in 143 Spanish HSCR patients. A total of four previously described polymorphisms and 12 novel sequence variants were detected. Of note, the novel R645C mutation was detected in 2 affected siblings of a HSCR family also carrying a RET splicing mutation. Using bioinformatics tools we observed that the presence of an additional cysteine residue might implicate structural alterations in the mutated protein. We propose haploinsufficiency as the most probable mechanism for the NTRK3 R645C mutation. NTRK3 and RET mutations in this family only appear together in the HSCR patients, suggesting that they per se are necessary but not sufficient to produce the phenotype. In addition, it is quite probable that the contribution of other still unidentified modifier genes, may be responsible for the different phenotypes (length of aganglionosis) in the two affected members.

Mutation Update of Spinal Muscular Atrophy in Spain: Molecular Characterization of 745 Unrelated Patients and Identification of Four Novel Mutations in the SMN1 Gene

Human Genetics. Feb, 2009  |  Pubmed ID: 19050931

Spinal muscular atrophy (SMA) is caused by mutations in the SMN1 gene. We have studied the molecular pathology of SMA in 745 unrelated Spanish patients using PCR-RFLP, SMN gene dosage analysis, linkage studies, long-range PCR and direct sequencing. Our systematic approach allowed us to complete genetic testing and risk assessment in 736 SMA patients (98.8%). Females were more frequently affected by the acute form of the disease (type I), whereas chronic forms (type II-III) predominated in males (p<0.008). Absence of the SMN1 gene was detected in 671 patients (90%), and hybrid SMN1-SMN2 genes were observed in 37 cases (5%). Furthermore, we detected 13 small mutations in 28 patients (3.8%), four of which were previously identified in other populations (c.91dupT; c.770_780dup11; p.Tyr272Cys and p.Thr274Ile), while five mutations were found to date only in Spanish patients (c.399_402delAGAG, p.Ile116Phe, p.Gln136Glu, c.740dupC and c.834+2T>G). The c.399_402delAGAG mutation accounted for 1.9% of all Spanish SMA patients. Finally, we discovered four novel mutations: c.312dupA, c.411delT, p.Trp190X and p.Met263Thr. Our results confirm that most SMA cases are due to large genetic rearrangements in the repetitive region of the SMA locus, resulting in absence-dysfunction of the SMN1 gene. By contrast, ancestrally inherited small mutations are responsible for only a small number of cases. Four prevalent changes in exons 3 and 6 (c.399_402delAGAG; c.770_780dup11; p.Tyr272Cys; p.Thr274Ile) accounted for almost 70% of our patients with these subtle mutations. An SMN-SMN dimer model featuring tight hydrophobic-aromatic interactions is proposed to explain the impact of mutations at the C-terminal end of the protein.

Endocytosis As a Mechanism of Regulating Natural Killer Cell Function: Unique Endocytic and Trafficking Pathway for CD94/NKG2A

Immunologic Research. 2009  |  Pubmed ID: 18979076

Natural killer (NK) cells are lymphocytes generally recognized as sentinels of the innate immune system due to their inherent capacity to deal with diseased (stressed) cells, including malignant and infected. This ability to recognize many potentially pathogenic situations is due to the expression of a diverse panel of activation receptors. Because NK cell activation triggers an aggressive inflammatory response, it is important to have a means of throttling this response. Hence, NK cells also express a panel of inhibitory receptors that recognize ligands expressed by "normal" cells. Little or nothing is known about the endocytosis and trafficking of NK cell receptors, which are of great relevance to understanding how NK cells maintain the appropriate balance of activating and inhibitory receptors on their cell surface. In this review, we focus on the ITIM-containing inhibitory receptor CD94/NKG2A showing that it is endocytosed by a previously undescribed macropinocytic-like process that may be related to the maintenance of its surface expression.

Lung Function and Clinical Risk Factors for Asthma in Infants and Young Children with Recurrent Wheeze

Thorax. Mar, 2009  |  Pubmed ID: 19008296

Although several risk factors for asthma have been identified in infants and young children with recurrent wheeze, the relevance of assessing lung function in this group remains unclear. Whether lung function is reduced during the first 2 years in recurrently wheezy children, with and without clinical risk factors for developing subsequent asthma (ie, parental asthma, personal history of allergic rhinitis, wheezing without colds and/or eosinophil level >4%) compared with healthy controls was assessed in this study.

Complexity of Phenotype-genotype Correlations in Spanish Patients with RDH12 Mutations

Investigative Ophthalmology & Visual Science. Mar, 2009  |  Pubmed ID: 19011012

Several mutations have been described in the RDH12 gene that disturb the activity of the encoded protein, suggesting that RDH12 loss of function disrupts the synthetic pathway of the visual chromophore 11-cis-retinal, therefore resulting in early and progressive retinal degeneration (RD). Mutations in this gene have been related to autosomal recessive Leber congenital amaurosis (LCA) and to a form of autosomal recessive childhood-onset severe retinal dystrophy (CSRD). This study was undertaken to attempt to correlate the genotype and phenotype in Spanish CSRD and LCA patients who harbor RDH12 mutations.

Endosomal Trafficking of the Ligated FcvarepsilonRI Receptor

Molecular Immunology. Feb, 2009  |  Pubmed ID: 18945491

In addition to initiating signaling cascades leading to mast cell mediator release, aggregation of the high affinity IgE receptor (FcvarepsilonRI) leads to rapid internalization of the cross-linked receptor. However, little is known about the trafficking of the internalized FcvarepsilonRI. Here we demonstrate that in RBL-2H3 cells, aggregated FcvarepsilonRI appears in the early endosomal antigen 1 (EEA1(+)) domains of the early endosomes within 15min after ligation. Minimal co-localization of FcvarepsilonRI with Rab5 was observed by 30min, followed by its appearance in the Rab7(+) late endosomes and lysosomes at later time points. During endosomal sorting, FcvarepsilonRIalpha and gamma subunits remain associated. In Syk-deficient RBL-2H3 cells, the rate of transport to lysosomes is markedly increased. Taken together, our data demonstrate time-dependent sorting of aggregated FcvarepsilonRI within the endosomal-lysosomal network, and that Syk may play an essential role in regulating the trafficking and retention of FcvarepsilonRI in endosomes.

Survival Probability of a Subdiffusive Particle in a D-dimensional Sea of Mobile Traps

Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics. Dec, 2009  |  Pubmed ID: 20365132

We investigate the long-time behavior of the survival probability P(t) of a mobile particle in d-dimensional continuous Euclidean media doped with noninteracting mobile traps. The particle is strictly subdiffusive, implying that its mean-square displacement grows as tgamma' with 0or=2 we find that at late times the survival probability is that of the pure target problem (the problem where the particle remains immobile) in agreement with previous studies for the d=1 case. These decay laws remain invariant over the whole gamma range as opposed to the dynamical crossover observed for the case of a purely diffusive particle (gamma'=1) where, for gamma<2/(2+d) , the survival probability becomes that of the so-called trapping problem (the problem where the particle moves in a sea of static traps). This behavior implies that for sufficiently low values of gamma(gamma<2/(2+d)) the survival probability becomes singular in the limit gamma'-->1: trappinglike for gamma'=1 and targetlike for any gamma'<1.

Is the RET Proto-oncogene Involved in the Pathogenesis of Intestinal Neuronal Dysplasia Type B?

Molecular Medicine Reports. Mar-Apr, 2009  |  Pubmed ID: 21475823

Hirschsprung disease (HSCR) is defined by the absence of intramural ganglia of Meissner and Auerbach along variable lengths of the gastrointestinal tract. Intestinal neuronal dysplasia (IND) type B is characterized by the malformation of the parasympathetic submucous plexus of the gut. A connection appears to exist between these two enteric nervous system abnormalities. Due to the major role played by the RET proto-oncogene in HSCR, we sought to determine whether this gene was also related to INDB. dHPLC techniques were employed to screen the RET coding region in 23 patients presenting with INDB and 30 patients with a combined HSCR+INDB phenotype. In addition, eight RET single nucleotide polymorphisms (SNPs) were strategically selected and genotyped by TaqMan technology. The distribution of SNPs and haplotypes was compared among the different groups of patients (INDB, HSCR+INDB, HSCR) and the controls. We found several RET mutations in our patients and some differences in the distribution of the RET SNPs among the groups of study. Our results suggest an involvement of RET in the pathogenesis of intestinal INDB, although by different molecular mechanisms than those leading to HSCR. Further investigation is warranted to elucidate these precise mechanisms and to clarify the genetic nature of INDB.

Mutation Spectrum of EYS in Spanish Patients with Autosomal Recessive Retinitis Pigmentosa

Human Mutation. Nov, 2010  |  Pubmed ID: 21069908

Retinitis pigmentosa (RP) is a heterogeneous group of inherited retinal dystrophies characterised ultimately by the loss of photoreceptor cells. We have recently identified a new gene(EYS) encoding an ortholog of Drosophila space maker (spam) as a commonly mutated gene in autosomal recessive RP. In the present study, we report the identification of 73 sequence variations in EYS, of which 28 are novel. Of these, 42.9% (12/28) are very likely pathogenic, 17.9% (5/28)are possibly pathogenic, whereas 39.3% (11/28) are SNPs. In addition, we have detected 3 pathogenic changes previously reported in other populations. We are also presenting the characterisation of EYS homologues in different species, and a detailed analysis of the EYS domains, with the identification of an interesting novel feature: a putative coiled-coil domain.Majority of the mutations in the arRP patients have been found within the domain structures of EYS. The minimum observed prevalence of distinct EYS mutations in our group of patients is of 15.9% (15/94), confirming a major involvement of EYS in the pathogenesis of arRP in the Spanish population. Along with the detection of three recurrent mutations in Caucasian population, our hypothesis of EYS being the first prevalent gene in arRP has been reinforced in the present study.

New Perspectives in Monitoring Drinking Water Microbial Quality

International Journal of Environmental Research and Public Health. Dec, 2010  |  Pubmed ID: 21318002

The safety of drinking water is evaluated by the results obtained from faecal indicators during the stipulated controls fixed by the legislation. However, drinking-water related illness outbreaks are still occurring worldwide. The failures that lead to these outbreaks are relatively common and typically involve preceding heavy rain and inadequate disinfection processes. The role that classical faecal indicators have played in the protection of public health is reviewed and the turning points expected for the future explored. The legislation for protecting the quality of drinking water in Europe is under revision, and the planned modifications include an update of current indicators and methods as well as the introduction of Water Safety Plans (WSPs), in line with WHO recommendations. The principles of the WSP approach and the advances signified by the introduction of these preventive measures in the future improvement of drinking water quality are presented. The expected impact that climate change will have in the quality of drinking water is also critically evaluated.

Divergent Series and Memory of the Initial Condition in the Long-time Solution of Some Anomalous Diffusion Problems

Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics. Feb, 2010  |  Pubmed ID: 20365528

We consider various anomalous d -dimensional diffusion problems in the presence of an absorbing boundary with radial symmetry. The motion of particles is described by a fractional diffusion equation. Their mean-square displacement is given by r(2) proportional, variant t(gamma)(00 , the emergence of such series in the long-time domain is a specific feature of subdiffusion problems. We present a method to regularize such series, and, in some cases, validate the procedure by using alternative techniques (Laplace transform method and numerical simulations). In the normal diffusion case, we find that the signature of the initial condition on the approach to the steady state rapidly fades away and the solution approaches a single (the main) decay mode in the long-time regime. In remarkable contrast, long-time memory of the initial condition is present in the subdiffusive case as the spatial part Psi1(r) describing the long-time decay of the solution to the steady state is determined by a weighted superposition of all spatial modes characteristic of the normal diffusion problem, the weight being dependent on the initial condition. Interestingly, Psi1(r) turns out to be independent of the anomalous diffusion exponent gamma .

Biomarkers in a Peat Deposit in Northern Spain (Huelga De Bayas, Asturias) As Proxy for Climate Variation

Journal of Chromatography. A. May, 2010  |  Pubmed ID: 20399440

Peatlands are peculiar ecosystems in which well-adapted communities grow and develop, recording the variation in climate and hydrological conditions inland. In addition necromass is well preserved and therefore peatlands can be used as palaeo-archives for environmental variation. In this work a peat core of depth 60 cm dated at the bottom of the peat deposit as ca. 250cal AD from Huelga de Bayas (Asturias, Northern Spain) was studied to a resolution of 2-4 cm to investigate the evolution of the environmental conditions in the area. Samples were extracted with a dichloromethane/methanol ratio of 3:1 and studied by means of gas chromatography (GC) and mass spectrometry (GC-MS) in order to identify possible biomarkers of climatic variation during the period of peat formation. Lipid biomarker study allows the identification of periods in which Sphagnum or higher plants preferentially contributed to the peat profile. The absolute dating of the profile combined with the n-alkane record displayed five episodes of wetter conditions around ca. 250 cal AD (Roman Warm Period), 1080 and 1270 cal AD (Medieval Warm Period), 1460 cal AD (Little Ice Age) and 1920 cal AD (Recent warming), which are consistent with climate evolution in the region. Pentacyclic triterpenoids with hopane skeleton derived from microorganisms and with oleanane skeleton derived from higher plants were identified. The presence of their ketone and acetyl-derivatives, along with the presence of unstable hopane configurations indicates a low maturity of the peat profile. A tendency for the functionalised triterpenoids to decrease with depth was observed in the profile.

Development and Characterization of Monoclonal Antibodies Against Rift Valley Fever Virus Nucleocapsid Protein Generated by DNA Immunization

MAbs. May-Jun, 2010  |  Pubmed ID: 20400862

This paper describes the generation of monoclonal antibodies directed to immunogenic nucleoprotein N epitopes of Rift Valley fever virus (RVFV), and their application in diagnostics, both for antibody detection in competitive ELISA and for antigen capture in a sandwich ELISA. Monoclonal antibodies (mAbs) were generated after DNA immunization of Balb/c mice and characterized by western blot, ELISA and cell immunostaining assays. At least three different immunorelevant epitopes were defined by mAb competition assays. Interestingly, two of the mAbs generated were able to distinguish between RVFV strains from Egyptian or South African lineages. These monoclonal antibodies constitute useful tools for diagnosis, especially for the detection of serum anti-RVFV antibodies from a broad range of species by means of competitive ELISA.

Novel MLPA Procedure Using Self-designed Probes Allows Comprehensive Analysis for CNVs of the Genes Involved in Hirschsprung Disease

BMC Medical Genetics. 2010  |  Pubmed ID: 20459765

Hirschsprung disease is characterized by the absence of intramural ganglion cells in the enteric plexuses, due to a fail during enteric nervous system formation. Hirschsprung has a complex genetic aetiology and mutations in several genes have been related to the disease. There is a clear predominance of missense/nonsense mutations in these genes whereas copy number variations (CNVs) have been seldom described, probably due to the limitations of conventional techniques usually employed for mutational analysis. In this study, we have looked for CNVs in some of the genes related to Hirschsprung (EDNRB, GFRA1, NRTN and PHOX2B) using the Multiple Ligation-dependent Probe Amplification (MLPA) approach.

HIV-2 Genetic Evolution in Patients with Advanced Disease is Faster Than That in Matched HIV-1 Patients

Journal of Virology. Jul, 2010  |  Pubmed ID: 20463072

The objective of this study was to estimate and compare the evolutionary rates of HIV-2 and HIV-1. Two HIV-2 data sets from patients with advanced disease were compared to matched HIV-1 data sets. The estimated mean evolutionary rate of HIV-2 was significantly higher than the estimated rate of HIV-1, both in the gp125 and in the V3 region of the env gene. In addition, the rate of synonymous substitutions in gp125 was significantly higher for HIV-2 than for HIV-1, possibly indicating a shorter generation time or higher mutation rate of HIV-2. Thus, the lower virulence of HIV-2 does not appear to translate into a lower rate of evolution.

Ischaemic Stroke in Incident Dialysis Patients

Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association. Oct, 2010  |  Pubmed ID: 20466665

Despite the high frequency of cardiovascular disease among the population on dialysis, there are few studies on ischaemic stroke and associated factors. The objective of the present study is to assess the prevalence of ischaemic stroke at the start of dialysis, its incidence in the course of follow-up and possible factors associated in its presentation.

A New Genotype of Lymphocystivirus Isolated from Cultured Gilthead Seabream, Sparus Aurata L., and Senegalese Sole, Solea Senegalensis (Kaup)

Journal of Fish Diseases. Aug, 2010  |  Pubmed ID: 20487141

Phosphorus Deficiency and Kinetics of Alkaline Phosphatase in Isolates and Natural Populations of Phototrophic Sulphur Bacteria

FEMS Microbiology Ecology. Aug, 2010  |  Pubmed ID: 20491922

Phosphorus deficiency was analysed in the oxic-anoxic gradient of the karstic sulphurous lakes Vilar and Sisó during the stratification period. The distribution of planktonic photosynthetic populations along a vertical gradient coincided with an increase in alkaline phosphatase activity (APA). A multiple stepwise correlation analysis of data yielded a positive correlation of APA with planktonic phototrophic populations. MUF-P hydrolysis saturation curves were used to estimate the enzyme kinetics. High-affinity phosphatases (i.e. low K(M) saturation constant) coincided with the oxic-anoxic gradient and progressively declined through both the epi- and the hypolimnion. Changes in the K(M) values are likely due to phosphate inhibition and the contribution of different planktonic populations in the induction of alkaline phosphatases. Extremely low organic phosphorus turnover times (as short as 0.37 h) were also estimated in the gradient zone, indicating a high dependence of the bacterial populations on organic phosphate esters. Phosphatase saturation kinetics revealed K(M) values from 0.53 to 8.45 microM MUF-P, perfectly matching those found in the isolates Thiocapsa sp. UdG3513, Chlorobium limicola UdG6050 and UdG6055 and Chlorobium phaeobacteroides CL1401. The results obtained indicate that a relevant adaptation of sulphur phototrophic bacteria may occasionally face periods of phosphate limitation despite thriving in nutrient-rich anoxic waters.

[Etanercept in Pregnancy and Breast-feeding]

Actas Dermo-sifiliográficas. May, 2010  |  Pubmed ID: 20492888

Etanercept is a biological drug, inhibitor of the Tumor Necrosis Factor indicated for the treatment of severe or moderate psoriasis resistant to other therapies. Because its use is becoming increasingly extended, we should know its possible teratogenic effects. The data provided by the literature and studies prior to the marketing of the product are very limited. Thus, it is necessary to study the presence of possible risks more through experiments in animal models and to conduct prolonged prospective studies in humans. With the current knowledge, it seems that suspension of the treatment from one month prior to becoming pregnant would provide an adequate safety margin, and that most of the patients who have become pregnant and have suspended etanercept as soon as they knew they were pregnant have not had any complications. However, the data needed to recommend etanercept for the control of psoriasis of a pregnant woman are very limited and controversial. Since the effects of a possible transfer of etanercept to maternal milk in a still-immature immune system are not known, in accordance with the risk/benefit principle, the use of etanercept should not be recommended in breast-feeding women.

Human Th1 Cells That Express CD300a Are Polyfunctional and After Stimulation Up-regulate the T-box Transcription Factor Eomesodermin

PloS One. 2010  |  Pubmed ID: 20498708

Human naïve CD4 T cells express low levels of the immunomodulatory receptor CD300a, whereas effector/memory CD4 cells can be either CD300a(+) or CD300a(-). This suggested that CD300a expression could define a specific subset within the effector/memory CD4 T cell subpopulations. In fact, ex vivo analysis of the IFN-gamma producing CD4 T cells showed that they are enriched in the CD300a(+) subset. Moreover, stimulated CD4 T cells producing TNF-alpha and IL-2 besides IFN-gamma (polyfunctional) are predominantly CD300a(+). In addition to producing markedly higher levels of Th1-associated cytokines, the stimulated CD300a(+) CD4 T cells are distinguished by a striking up-regulation of the T-box transcription factor eomesodermin (Eomes), whereas T-bet is up-regulated in both CD300a(+) and CD300a(-) activated CD4 T cells to similar levels. The pleiotropic cytokine TGF-beta1 has a determinant role in dictating the development of this Th1 subset, as its presence inhibits the expression of CD300a and down-regulates the expression of Eomes and IFN-gamma. We conclude that CD300a(+) human Th1 cells tend to be polyfunctional and after stimulation up-regulate Eomes.

[Safety of Azathioprine Therapy Adjusted to Thiopurine Methyltransferase Activity in the Treatment of Infantile Atopic Dermatitis. Report on 7 Cases]

Actas Dermo-sifiliográficas. Jun, 2010  |  Pubmed ID: 20525484

In a small number of cases of childhood atopic dermatitis, topical therapy is ineffective, necessitating prolonged use of systemic immunosuppressants. Over the last few years, a better understanding of the metabolic pathways involved in azathioprine breakdown has enabled us to use this drug more safely. In this study, we evaluated the toxicity of azathioprine treatment adjusted to thiopurine methyltransferase activity in children with severe atopic dermatitis.

The C.859G>C Variant in the SMN2 Gene is Associated with Types II and III SMA and Originates from a Common Ancestor

Journal of Medical Genetics. Sep, 2010  |  Pubmed ID: 20577007

Homozygous mutations of the telomeric SMN1 gene lead to degeneration of motor neurons causing spinal muscular atrophy (SMA). A highly similar centromeric gene (SMN2) can only partially compensate for SMN1 deficiency. The c.859G>C variant in SMN2 has been recently reported as a positive disease modifier. We identified the variant in 10 unrelated chronic SMA patients with a wide spectrum of phenotypes ranging from type II patients who can only sit to adult walkers. Haplotype analysis strongly suggests that the variant originated from a common ancestor. Our results confirm that the c.859G>C variant is a milder SMN2 allele and predict a direct correlation between SMN activity and phenotypic severity.

Monitoring of Firefighters Exposure to Smoke During Fire Experiments in Portugal

Environment International. Oct, 2010  |  Pubmed ID: 20579737

Forest fires represent a serious threat to public security in Europe due to the large burned area. Moreover, smoke pollution due to forest fire events is an important public health issue for the communities directly affected, and particularly for the personnel involved in firefighting operations. Aiming to contribute to the scientific knowledge concerning firefighters exposure to forest fires smoke, data of individual exposure to carbon monoxide, nitrogen dioxide, volatile organic compounds, and particulate matter were obtained during experimental field fires for a group of 10 firefighters equipped with portable "in continuum" measuring devices. Measured values are very high exceeding the Occupational Exposure Standard limits, in particular for peak limit thresholds. These are the first measurements and analysis of firefighter's individual exposure to toxic gases and particles in fire smoke experiments in Europe. However, they already indicate that urgent measures to avoid these levels of exposure are needed.

[Scarring Alopecia in a Black Patient]

Actas Dermo-sifiliográficas. Jan-Feb, 2010  |  Pubmed ID: 20109397

Physicochemical Characterization of Silylated Functionalized Materials

Journal of Colloid and Interface Science. Apr, 2010  |  Pubmed ID: 20129614

Silylation of several materials where the surface area arises from the internal pores (MCM-41 and FSM-16) or is essentially external (silica gel, and clays) was performed using three organosilanes: (3-aminopropyl)triethoxysilane (APTES), 4-(triethoxysilyl)aniline (TESA) and (3-mercaptopropyl)trimethoxysilane (MPTS). The materials were characterized by nitrogen adsorption-desorption at -196 degrees C, powder XRD, XPS, bulk chemical analysis, FTIR and (29)Si and (13)C MAS NMR. For MCM-41 and FSM-16 the highest amounts of organosilane are obtained for APTES, while for the remaining materials the highest amounts are for MPTS; TESA always anchored with the lowest percentage. In terms of surface chemical analysis, TESA anchored with the highest contents irrespectively of the material, and the opposite is registered for MPTS. Comparison of bulk vs surface contents indicate that TESA is mainly anchored at the material external surface. Moreover, with N or S (surface and bulk) contents expressed per unit of surface area, MCM-41 and FSM-16 (internal porosity) show the lowest amounts of silane; the highest amounts of silane per unit of surface area are obtained for the clays. Grafting of the organosilanes to the surface hydroxyl groups was corroborated by FTIR and (29)Si and (13)C MAS NMR. Furthermore, NMR data suggested that TESA and APTES grafted mostly through a bidentate approach, whereas MPTS grafted by a monodentate mechanism.

Involvement of SOX10 in the Pathogenesis of Hirschsprung Disease: Report of a Truncating Mutation in an Isolated Patient

Journal of Molecular Medicine (Berlin, Germany). May, 2010  |  Pubmed ID: 20130826

SOX10 protein is a key transcription factor during neural crest development. Mutations in SOX10 are associated with several neurocristopathies such as Waardenburg syndrome type IV (WS4), a congenital disorder characterized by the association of hearing loss, pigmentary abnormalities, and absence of ganglion cells in the myenteric and submucosal plexus of the gastrointestinal tract, also known as aganglionic megacolon or Hirschsprung disease (HSCR). Several mutations at this locus are known to cause a high percentage of WS4 cases, but no SOX10 mutations had been ever reported associated to isolated HSCR patient. Therefore, nonsyndromic HSCR was initially thought not to be associated to mutations at this particular locus. In the present study, we describe the evaluation of the SOX10 gene in a series of 196 isolated HSCR cases, the largest patient series evaluated so far, and report a truncating c.153-155del mutation. This is the first time that a SOX10 mutation is detected in an isolated HSCR patient, which completely changes the scenario for the implications of SOX10 mutations in human disease, giving us a new tool for genetic counseling.

New Mutations in BBS Genes in Small Consanguineous Families with Bardet-Biedl Syndrome: Detection of Candidate Regions by Homozygosity Mapping

Molecular Vision. 2010  |  Pubmed ID: 20142850

Bardet-Biedl syndrome (BBS, OMIM 209900) is a rare multi-organ disorder in which BBS patients manifest a variable phenotype that includes retinal dystrophy, polydactyly, mental delay, obesity, and also reproductive tract and renal abnormalities. Mutations in 14 genes (BBS1-BBS14) are found in 70% of the patients, indicating that additional mutations in known and new BBS genes remain to be identified. Therefore, the molecular diagnosis of this complex disorder is a challenging task.

Frequency Analysis of Air Quality Time Series for Traffic Related Pollutants

Journal of Environmental Monitoring : JEM. Feb, 2010  |  Pubmed ID: 20145898

In the present work, annual time series of traffic related pollutants (CO and PM(10)) were considered for frequency analysis (Fourier series) with the aim to understand the underlying physical processes and the influence of emission sources on the variability of the air pollutant concentrations. Several urban traffic and suburban background air quality stations located in Porto metropolitan area (Portugal) were analysed. The results obtained for CO and PM(10) reveal the important contributions of short-term fluctuations (12 h and 24 h periods). However, the spectrum signals at low frequencies are significantly different between these pollutants thus stressing that temporal variations of CO and PM(10) are influenced by different processes. Cross-spectrum analysis of the air quality time series against wind measurements and traffic counts allowed us to identify the contribution of long-range transport over a period of about 21 days to the PM(10) fluctuations. Also, a correlation of over 80% between the pollution levels in the vicinity of traffic sources and suburban background levels are found for these harmonic components in the PM(10) spectrum, while correlations for CO is below a significant level. Thus, the spectrum and cross-spectrum analysis performed in this study reveal the distinct influence of local traffic emissions and long-range transport to CO and PM(10) fluctuations in the polluted urban area. The methodology shows to be a powerful tool for the analysis of the causes of air pollution.

Effects of Cultural Conditions on Fungal Biomass, Blastospore Yields and Toxicity of Fungal Secreted Proteins in Batch Cultures of Metarhizium Anisopliae (Ascomycota: Hypocreales)

Pest Management Science. Jul, 2010  |  Pubmed ID: 20232285

Recently, two fungal proteins with apparent molecular masses of 11 and 15 kDa and insecticidal activity against Ceratitis capitata (Wied.) have been purified from the crude soluble protein extract (CSPE) secreted by the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin (strain EAMa 01/58-Su) in Adamek's liquid medium. The feasibility of culturing this strain in fermentation facilities in order to harvest and formulate the insecticidal proteins for C. capitata control is mainly dependent on the ability to produce high concentrations of the active proteins at a reasonable cost.

Identification of Novel Mutations in the Ortholog of Drosophila Eyes Shut Gene (EYS) Causing Autosomal Recessive Retinitis Pigmentosa

Investigative Ophthalmology & Visual Science. Aug, 2010  |  Pubmed ID: 20237254

Recently, a novel gene was cloned for autosomal recessive retinitis pigmentosa (arRP), EYS, on 6q12. This study was conducted to determine the spectrum and frequency of EYS mutations in 195 unrelated patients with autosomal recessive and autosomal dominant RP (adRP).

The Management of the Allergic Child at School: EAACI/GA2LEN Task Force on the Allergic Child at School

Allergy. Jun, 2010  |  Pubmed ID: 20345502

Allergy affects at least one-quarter of European schoolchildren, it reduces quality of life and may impair school performance; there is a risk of severe reactions and, in rare cases, death. Allergy is a multi-system disorder, and children often have several co-existing diseases, i.e. allergic rhinitis, asthma, eczema and food allergy. Severe food allergy reactions may occur for the first time at school, and overall 20% of food allergy reactions occur in schools. Up to two-thirds of schools have at least one child at risk of anaphylaxis but many are poorly prepared. A cooperative partnership between doctors, community and school nurses, school staff, parents and the child is necessary to ensure allergic children are protected. Schools and doctors should adopt a comprehensive approach to allergy training, ensuring that all staff can prevent, recognize and initiate treatment of allergic reactions.

Cystic Fibrosis, Atopy, Asthma and ABPA

Allergologia Et Immunopathologia. Sep-Oct, 2010  |  Pubmed ID: 20675033

The role of atopy on cystic fibrosis (CF) progression remains unclear but evidence suggests that it may influence the appearance of co-morbid conditions such as CF asthma or allergic bronchopulmonary aspergillosis (ABPA). Recognising asthma in patients with CF is not always easy but the identification of atopic markers favours the diagnosis. Physicians should be aware of this fact in order to achieve a better control of respiratory symptoms in patients with CF. Bronchial mucosa inflammation and abnormal mucus predispose to mould colonisation. These patients are at higher risk of allergic sensitisation, especially when atopic susceptibility is present. In the particular case of A. fumigatus, allergic sensitisation precedes ABPA development, which occurs in up to 10% of CF patients. Progression of lung function deterioration is most strikingly pronounced in patients with ABPA. Therefore, sensitisation with A. fumigatus should be regularly tested in patients with CF, especially those at higher risk. Recombinant allergens constitute an important advance in differentiating Aspergillus sensitisation from ABPA itself.

Potent and Broadly Reactive HIV-2 Neutralizing Antibodies Elicited by a Vaccinia Virus Vector Prime-C2V3C3 Polypeptide Boost Immunization Strategy

Journal of Virology. Dec, 2010  |  Pubmed ID: 20844029

Human immunodeficiency virus type 2 (HIV-2) infection affects about 1 to 2 million individuals, the majority living in West Africa, Europe, and India. As for HIV-1, new strategies for the prevention of HIV-2 infection are needed. Our aim was to produce new vaccine immunogens that elicit the production of broadly reactive HIV-2 neutralizing antibodies (NAbs). Native and truncated envelope proteins from the reference HIV-2ALI isolate were expressed in vaccinia virus or in bacteria. This source isolate was used due to its unique phenotype combining CD4 independence and CCR5 usage. NAbs were not elicited in BALB/c mice by single immunization with a truncated and fully glycosylated envelope gp125 (gp125t) or a recombinant polypeptide comprising the C2, V3, and C3 envelope regions (rpC2-C3). A strong and broad NAb response was, however, elicited in mice primed with gp125t expressed in vaccinia virus and boosted with rpC2-C3. Serum from these animals potently neutralized (median 50% neutralizing titer, 3,200) six of six highly divergent primary HIV-2 isolates. Coreceptor usage and the V3 sequence of NAb-sensitive isolates were similar to that of the vaccinating immunogen (HIV-2ALI). In contrast, NAbs were not reactive on three X4 isolates that displayed major changes in V3 loop sequence and structure. Collectively, our findings demonstrate that broadly reactive HIV-2 NAbs can be elicited by using a vaccinia virus vector-prime/rpC2-C3-boost immunization strategy and suggest a potential relationship between escape to neutralization and cell tropism.

Novel Association of Severe Neonatal Encephalopathy and Hirschsprung Disease in a Male with a Duplication at the Xq28 Region

BMC Medical Genetics. 2010  |  Pubmed ID: 20860806

Hirschsprung disease (HSCR) is a neurocristopathy characterized by the absence of parasympathetic intrinsic ganglion cells in the submucosal and myenteric plexuses along a variable portion of the intestinal tract. In approximately 18% of the cases HSCR also presents with multiple congenital anomalies including recognized syndromes.

Genotypes and Antimicrobial-resistant Phenotypes of Neisseria Gonorrhoeae in Portugal (2004-2009)

Sexually Transmitted Infections. Nov, 2010  |  Pubmed ID: 20940158

To determine the antibiotic phenotype and MAST-genotype distribution of Neisseria gonorrhoeae isolates in Portugal between 2004 and 2009, and to evaluate specific associations between MAST-genotypes and sexual orientation, age and antibiotic resistance.

Adaptive Evolution of the Chlamydia Trachomatis Dominant Antigen Reveals Distinct Evolutionary Scenarios for B- and T-cell Epitopes: Worldwide Survey

PloS One. 2010  |  Pubmed ID: 20957150

Chlamydia trachomatis is one of the most disseminated human pathogens, for which no vaccine is available yet. Understanding the impact of the host pressure on pathogen antigens is crucial, but so far it was only assessed for highly-restricted geographic areas. We aimed to evaluate the evolutionary picture of the chlamydial key antigen (MOMP), which is one of the leading multi-subunit vaccine candidates, in a worldwide basis.

Renal Transplant Register of Andalusia, 2010 Report: Survival in Relation to the Factors Used in Recipient Selection

Transplantation Proceedings. Oct, 2010  |  Pubmed ID: 20970627

Kidney transplantation is the best therapeutic alternative for patients suffering from end-stage renal disease (ESRD). In recent years a significant advance has been made in Andalusia in graft and recipient survival as seen in our 2009 publication. In the current work we analyzed 2989 kidney transplantations performed between January 1, 2000 and December 31, 2009 based on data obtained from the Renal Transplant Registry of Andalusia. We studied the influence on graft and patient survival of factors, such as donor and recipient age, HLA matching, HLA immunization, and duration of previous renal replacement therapy. Patient survival was influenced by age at the time of transplantation and by donor age; the younger the donor, the more it was improved. Graft survival was determined by the donor age group, with no differences at each level according to the recipient age group. No significant differences were observed in patient survival or graft or death-censored graft survival according to HLA matching. Patient and graft survivals were significantly affected by the duration of the previous renal replacement therapy. Despite this being a preliminary study, we have shown the importance of nonmodifiable factors in transplant survival, such as donor and recipient age, with HLA matching having a limited effect. These latter findings should be confirmed in the future by multivariate analyses.

Toxicity of Photobacterium Damselae Subsp. Damselae Strains Isolated from New Cultured Marine Fish

Diseases of Aquatic Organisms. Oct, 2010  |  Pubmed ID: 21166312

The in vivo and in vitro toxicity of bacterial cells and their extracellular products (ECPs) from 16 strains of Photobacterium damselae subsp. damselae isolated from 7 epizootic outbreaks were evaluated. On the basis of their 50% lethal dose (LD50) values (about 1 x 10(50 CFU), these strains may be considered as moderately virulent. However, their ECPs were strongly lethal for redbanded seabream Pagrus auriga causing fish death within 2 h post-inoculation (protein concentration ranged between 2.1 and 6.41 microg g(-1) fish). The bacterial ECPs tested exhibited several enzymatic activities, such as amylase, lipase, phospholipase, alkaline phosphatase, esterase-lipase, acid phosphatase, and beta-glucosaminidase. These ECPs displayed a strong cytotoxic effect on 4 fish and 2 mammalian cell lines, although this activity disappeared when ECPs were heated at 100 degrees C. The virulence of the strains tested could not be related to the hemolytic activity or to the production of the toxin damselysin. Therefore, another unknown type of toxin could play an important role in the virulence mechanisms of this bacterial pathogen.

Are the Defined Substrate-based Methods Adequate to Determine the Microbiological Quality of Natural Recreational Waters?

Journal of Water and Health. Mar, 2010  |  Pubmed ID: 20009243

Monitoring the microbiological quality of water used for recreational activities is very important to human public health. Although the sanitary quality of recreational marine waters could be evaluated by standard methods, they are time-consuming and need confirmation. For these reasons, faster and more sensitive methods, such as the defined substrate-based technology, have been developed. In the present work, we have compared the standard method of membrane filtration using Tergitol-TTC agar for total coliforms and Escherichia coli, and Slanetz and Bartley agar for enterococci, and the IDEXX defined substrate technology for these faecal pollution indicators to determine the microbiological quality of natural recreational waters. ISO 17994:2004 standard was used to compare these methods. The IDEXX for total coliforms and E. coli, Colilert, showed higher values than those obtained by the standard method. Enterolert test, for the enumeration of enterococci, showed lower values when compared with the standard method. It may be concluded that more studies to evaluate the precision and accuracy of the rapid tests are required in order to apply them for routine monitoring of marine and freshwater recreational bathing areas. The main advantages of these methods are that they are more specific, feasible and simpler than the standard methodology.

New Roles of EDNRB and EDN3 in the Pathogenesis of Hirschsprung Disease

Genetics in Medicine : Official Journal of the American College of Medical Genetics. Jan, 2010  |  Pubmed ID: 20009762

Hirschsprung disease is characterized by the absence of intramural ganglion cells in the myenteric and submucosal plexuses within distal intestine, because of a fail in the enteric nervous system formations process. Endothelin-3-endothelin receptor B signaling pathway is known to play an essential role in this process. The aim of this study was to evaluate the implication of the EDN3 and EDNRB genes in a series of patients with Hirschsprung disease from Spain and determinate their mutational spectrum.

A Combined RT-PCR and Dot-blot Hybridization Method Reveals the Coexistence of SJNNV and RGNNV Betanodavirus Genotypes in Wild Meagre (Argyrosomus Regius)

Journal of Applied Microbiology. Oct, 2010  |  Pubmed ID: 20497277

To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study.

Normalization Strategies for Real-time Expression Data in Chlamydia Trachomatis

Journal of Microbiological Methods. Sep, 2010  |  Pubmed ID: 20619305

Chlamydia trachomatis is a widespread obligate intracellular pathogen genetically non-tractable for which transcriptomics is a fundamental tool to better understand its biology. However, the suitability of endogenous controls for normalization of transcriptomic data in this bacterium still needs validation. We aimed to assess the stability of 10 genes for their potential use as endogenous controls in real-time quantitative PCR assays at both normal and stress (D-cycloserine treatment) growth conditions throughout the developmental cycle of three C. trachomatis strains with different tissue tropism. Normalization was performed by real-time absolute quantification of the bacterial genomes. We also tested the applicability of two widely used softwares (geNorm and Normfinder) to our data. For all strains, we found that 16SrRNA was the most stably expressed gene throughout the chlamydial normal developmental cycle, which indicates its potential use as endogenous control in relative expression assays. However, it was highly unstable under D-cycloserine treatment (where oppA_2 was top-ranked), suggesting prudence when using ribosomal genes in expression experiments involving stress conditions. The geNorm and Normfinder algorithms revealed contrasting results and seem inappropriate for the selected pool of genes. Considering the multiplicity of experimental conditions, there should be an in loco validation of endogenous controls, where 16SrRNA appears to be in the front line. Alternatively, normalization of expression data against genomic DNA, which is less influenced by experimental constraints that are especially relevant for intracellular organisms, likely constitutes a good option. Moreover, the number of genomes also seems to be less subject to variation than expression of endogenous controls when working under stress conditions. The present study constitutes the first evaluation of putative endogenous controls for real-time expression assays in C. trachomatis.

Vertical Distribution of Ammonia-oxidizing Crenarchaeota and Methanogens in the Epipelagic Waters of Lake Kivu (Rwanda-Democratic Republic of the Congo)

Applied and Environmental Microbiology. Oct, 2010  |  Pubmed ID: 20802065

Four stratified basins in Lake Kivu (Rwanda-Democratic Republic of the Congo) were sampled in March 2007 to investigate the abundance, distribution, and potential biogeochemical role of planktonic archaea. We used fluorescence in situ hybridization with catalyzed-reported deposition microscopic counts (CARD-FISH), denaturing gradient gel electrophoresis (DGGE) fingerprinting, and quantitative PCR (qPCR) of signature genes for ammonia-oxidizing archaea (16S rRNA for marine Crenarchaeota group 1.1a [MCG1] and ammonia monooxygenase subunit A [amoA]). Abundance of archaea ranged from 1 to 4.5% of total DAPI (4',6-diamidino-2-phenylindole) counts with maximal concentrations at the oxic-anoxic transition zone (∼50-m depth). Phylogenetic analysis of the archaeal planktonic community revealed a higher level of richness of crenarchaeal 16S rRNA gene sequences (21 of the 28 operational taxonomic units [OTUs] identified [75%]) over euryarchaeotal ones (7 OTUs). Sequences affiliated with the kingdom Euryarchaeota were mainly recovered from the anoxic water compartment and mostly grouped into methanogenic lineages (Methanosarcinales and Methanocellales). In turn, crenarchaeal phylotypes were recovered throughout the sampled epipelagic waters (0- to 100-m depth), with clear phylogenetic segregation along the transition from oxic to anoxic water masses. Thus, whereas in the anoxic hypolimnion crenarchaeotal OTUs were mainly assigned to the miscellaneous crenarchaeotic group, the OTUs from the oxic-anoxic transition and above belonged to Crenarchaeota groups 1.1a and 1.1b, two lineages containing most of the ammonia-oxidizing representatives known so far. The concomitant vertical distribution of both nitrite and nitrate maxima and the copy numbers of both MCG1 16S rRNA and amoA genes suggest the potential implication of Crenarchaeota in nitrification processes occurring in the epilimnetic waters of the lake.

Molecular Characterization and Antimicrobial Susceptibility Profiles in Streptococcus Agalactiae Colonizing Strains: Association of Erythromycin Resistance with Subtype III-1 Genetic Clone Family

Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases. Sep, 2010  |  Pubmed ID: 19886900

Knowledge of the epidemiology of Streptococcus agalactiae in Portugal is limited: therefore, the present study aimed to investigate the carriage rate of S. agalactiae among Portuguese women of reproductive age and the prevalence of antibiotic resistance, as well as to perform a molecular characterization of the clinical isolates. S. agalactiae was recovered from 6.2% of 4269 women during the period 2005–2007, with a predominance of capsular genotypes III (35%), V (33%), Ia (16%) and II (10%) in a sample of 100 isolates. To our knowledge, this is the first report of the S. agalactiae colonization rate in Portugal determined according to CDC guidelines. All isolates were susceptible to penicillin and vancomycin, whereas resistance to clindamycin and erythromycin was detected in 10% and 19% of isolates, respectively. Among the 19 erythromycin-resistant isolates, ten (53%) displayed the constitutive MLS(B) phenotype (conferring high-level resistance to macrolides), eight (42%) had the inducible MLS(B), and the M phenotype accounted for one isolate (5%). erm methylase genes were exclusively associated with MLS(B) phenotype isolates, whereas the M phenotype was a result of the presence of mefA. Multilocus sequence typing analysis of the genetic relatedness among isolates presenting resistance to erythromycin demonstrated a novel association between erythromycin resistance and the subtype III-1/ST-19 genetic clone family.

Detection of Infectious Pancreatic Necrosis Virus (IPNV) from Asymptomatic Redbanded Seabream, Pagrus Auriga Valenciennes, and Common Seabream, Pagrus Pagrus (L.), Using a Non-destructive Procedure

Journal of Fish Diseases. Apr, 2010  |  Pubmed ID: 20059634

A non-destructive procedure based on nested RT-PCR and dot-blot hybridization has been developed for the detection of asymptomatic IPNV-carrier fish. The pair of primers designed for RT-PCR amplified a 599-bp fragment of the pVP2 region within the polyprotein gene, resulting in the detection of IPNV genotype III.1. The use of a nested RT-PCR allowed the amplification of IPNV genotypes III.1 and I.2. In addition, a 191-bp probe was designed for hybridization studies used in combination with the nested RT-PCR. The application of the nested RT-PCR to analyse blood samples from asymptomatic redbanded seabream, Pagrus auriga, and common seabream, P. pagrus, specimens showed a 53.1% and 77.8% prevalence of IPNV-carriers, respectively. The combination of nested RT-PCR and dot-blot hybridization increased the detection rates up to 100% for redbanded seabream and 94.4% for common seabream. Therefore, the protocol described in this study is highly sensitive and specific for the detection of IPNV in asymptomatic carrier fish, and, in addition, the results demonstrate the carrier state in two newly cultured sparid species in southern Spain.

RNA Immunization Can Protect Mice Against Foot-and-mouth Disease Virus

Antiviral Research. Mar, 2010  |  Pubmed ID: 20005905

In previous work we have reported the immunization of swine using in vitro-transcribed foot-and-mouth disease virus (FMDV) RNA. With the aim of testing whether RNA-induced immunization can mediate protection against viral infection, a group of Swiss adult mice was inoculated with FMDV infectious transcripts. In most inoculated animals viral RNA was detected in serum at 48-72h postinoculation. A group of the RNA-inoculated mice (11 out of 19) developed significant titers of neutralizing antibodies against FMDV. Among those animals that were successfully challenged with infectious virus (15 out of 19), three out of the eight animals immunized upon RNA inoculation were protected, as infectious virus could not be isolated from sera but specific anti-FMDV antibodies could be readily detected. These results suggest the potential of the inoculation of genetically engineered FMDV RNA for virulence and protection assays in the murine model and allow to explore the suitability of RNA-based FMDV vaccination in natural host animals.

Cellular and Molecular Immune Responses of the Sea Bass (Dicentrarchus Labrax) Experimentally Infected with Betanodavirus

Fish & Shellfish Immunology. Feb, 2010  |  Pubmed ID: 19925869

Naïve sea bass juveniles (38.4 + or - 4.5 g) were intramuscularly infected with a sublethal dose of betanodavirus isolate 378/I03, followed after 43 days by a similar boosting. This infection resulted in an overall mortality of 7.6%. At various intervals, sampling of fish tissues was performed to investigate: i) B and T lymphocyte content in organs and tissues; ii), proliferation of leucocytes re-stimulated in vitro with inactivated virus; iii) presence of serum antibody specific for betanodavirus; iv) expression of genes coding for the following immunoregulatory molecules involved in innate and acquired responses: type I IFN, Mx, IL-1, Cox-2; IL-10, TGF-beta, TCRbeta, CD4, CD8alpha, IgM, by using a quantitative PCR array system developed for sea bass. The obtained results showed a detectable increase of T cells and B cells in PBL during betanodavirus infection. Furthermore, leucocytes obtained from blood, head kidney, and gills showed a detectable "in vitro" increase in viability upon addition of inactivated viral particles, as determined by measuring intracellular ATP concentration. ELISA analysis of sera showed that exposure to nodavirus induced a low, but specific antibody titer measured 43 days after infection, despite the presence of measurable levels of natural antibody. Finally, a strong upregulation of genes coding for type I IFN, Mx, and IgM was identified after both infection and boosting. Interestingly, an upregulation of Cox-2 until boosting, and of TGF-beta and IL-10 after boosting was also observed, while the other tested genes did not show any significant variations with respect to mock-treated fish. Overall, our work represents a first comprehensive analysis of cellular and molecular immune parameters in a fish species exposed to a pathogenic virus.

Molecular Intraspecific Characterization of Photobacterium Damselae Ssp. Damselae Strains Affecting Cultured Marine Fish

Journal of Applied Microbiology. Jun, 2010  |  Pubmed ID: 19922594

The aim of this study was to analyse the intraspecific variability of Photobacterium damselae ssp. damselae strains isolated from different cultured marine fish species using molecular typing methods.

One-step Multiplex Polymerase Chain Reaction for Preimplantation Genetic Diagnosis of Huntington Disease

Fertility and Sterility. May, 2010  |  Pubmed ID: 19423106

To develop a multiplex polymerase chain reaction (PCR) method for Huntington disease (HD) preimplantation genetic diagnosis (PGD) based on the coamplification of CAG repeats and three different polymorphic microsatellites in a single step of PCR.

Differential Contributions of Rare and Common, Coding and Noncoding Ret Mutations to Multifactorial Hirschsprung Disease Liability

American Journal of Human Genetics. Jul, 2010  |  Pubmed ID: 20598273

The major gene for Hirschsprung disease (HSCR) encodes the receptor tyrosine kinase RET. In a study of 690 European- and 192 Chinese-descent probands and their parents or controls, we demonstrate the ubiquity of a >4-fold susceptibility from a C-->T allele (rs2435357: p = 3.9 x 10(-43) in European ancestry; p = 1.1 x 10(-21) in Chinese samples) that probably arose once within the intronic RET enhancer MCS+9.7. With in vitro assays, we now show that the T variant disrupts a SOX10 binding site within MCS+9.7 that compromises RET transactivation. The T allele, with a control frequency of 20%-30%/47% and case frequency of 54%-62%/88% in European/Chinese-ancestry individuals, is involved in all forms of HSCR. It is marginally associated with proband gender (p = 0.13) and significantly so with length of aganglionosis (p = 7.6 x 10(-5)) and familiality (p = 6.2 x 10(-4)). The enhancer variant is more frequent in the common forms of male, short-segment, and simplex families whereas multiple, rare, coding mutations are the norm in the less common and more severe forms of female, long-segment, and multiplex families. The T variant also increases penetrance in patients with rare RET coding mutations. Thus, both rare and common mutations, individually and together, make contributions to the risk of HSCR. The distribution of RET variants in diverse HSCR patients suggests a "cellular-recessive" genetic model where both RET alleles' function is compromised. The RET allelic series, and its genotype-phenotype correlations, shows that success in variant identification in complex disorders may strongly depend on which patients are studied.

Distinct Early Inflammatory Events During Ear Tissue Regeneration in Mice Selected for High Inflammation Bearing Slc11a1 R and S Alleles

Inflammation. Oct, 2011  |  Pubmed ID: 20665098

High inflammatory AIRmax mice homozygous for Slc11a1 R and S alleles were produced. AIRmax(SS) mice showed faster ear tissue regeneration than AIRmax(RR) mice, suggesting that the S allele favored tissue restoration. Here, we investigated the gene expression profiles and the inflammatory reactions of AIRmax(RR) and AIRmax(SS) mice during the initial phase of ear tissue regeneration. We observed superior levels of analysis of wound myeloperoxidase and edema in AIRmax(SS) mice, although similar cell influx was verified in both lines. Of the genes, 794 were up- and 674 down-regulated in AIRmax(RR), while 735 genes were found to be up- and 1616 down-regulated in AIRmax(SS) mice 48 h after punch. Both mouse lines showed significant over-represented genes related to cell proliferation; however AIRmax(SS) displayed up-regulation of inflammatory response genes. Quantitative PCR experiments showed higher expressions of Tgfb1, Dap12 and Trem1 genes in AIRmax(SS) mice. These results indicate that Slc11a1 gene modulated the early inflammatory events of ear tissue regeneration.

Three Mx Genes with Differential Response to VNNV Infection Have Been Identified in Gilthead Seabream (Sparus Aurata)

Molecular Immunology. May, 2011  |  Pubmed ID: 21466896

Type I interferons are secreted by infected cells and promote an antiviral state in neighbouring cells by the induction of numerous genes, some of which present antiviral activity, as the Mx proteins. In this study, three different Mx cDNAs (Mx1, Mx2 and Mx3) from gilthead seabream (Sparus aurata), the most important fish species in Southern European aquaculture, have been cloned and characterized. A Southern blot assay revealed the existence of three Mx loci, thus the three Mx isoforms correspond to three different genes that seem to have a common origin. The genomic sequences of Mx1, Mx2 and Mx3 have been completely obtained, and consist on 11 introns and 12 exons in a full length of 5971 bp, 7391 bp and 6938 bp, respectively. As a first approach to the functional meaning of these three genes, their response to the viral nervous necrosis virus (VNNV) infection was tested. Important differences in terms of tissue, time course and level of induction were found between them, thus suggesting a differential functional role for each isoform, which can represent a key point in the natural resistance of this fish species, that has been repeatedly reported as an asymptomatic carrier of VNNV.

Molecular Epidemiology of Group B Streptococcal Meningitis in Children Beyond the Neonatal Period from Angola

Journal of Medical Microbiology. Sep, 2011  |  Pubmed ID: 21474607

Streptococcus agalactiae is a major pathogen of neonates and immunocompromised adults. Prior studies have demonstrated that, beyond the neonatal period, S. agalactiae rarely causes invasive infections in children. However, during 2004-2005, S. agalactiae was the causative agent of 60 meningitis episodes in children aged 3 months to 12 years from Angola. To identify and study the specific causative genetic lineages of S. agalactiae childhood meningitis, which lack characterization to date, we conducted an extensive molecular analysis of the recovered isolates (n = 21). This constitutes what we believe to be the first molecular study of the population structure of invasive S. agalactiae isolates from Africa. A low genetic diversity was observed among the isolates, where the majority belonged to clonal complex (CC) 17 presenting the capsular subtype III-2 (86 % of cases) and marked by the intron group II GBSi1, which has previously been observed to be associated with neonatal hosts. The predominance of single-locus variants of sequence type (ST) 17 suggested the local diversification of this hypervirulent clone, which displayed novel alleles of the fbsB and sip virulence genes. The absence of the scpB-lmb region in two S. agalactiae isolates with the Ia/ST23 genotype is more typical of cattle than human isolates. Globally, these data provide novel information about the enhanced invasiveness of the CC17 genetic lineage in older children and suggest the local diversification of this clone, which may be related to the future emergence of a novel epidemic clone in Angola.

Skin Dose Mapping for Fluoroscopically Guided Interventions

Medical Physics. Oct, 2011  |  Pubmed ID: 21992367

To introduce a new skin dose mapping software system for interventional fluoroscopy dose assessment and to analyze the benefits and limitations of patient-phantom matching.

Comprehensive Analysis of RET Common and Rare Variants in a Series of Spanish Hirschsprung Patients Confirms a Synergistic Effect of Both Kinds of Events

BMC Medical Genetics. 2011  |  Pubmed ID: 21995290

RET is the major gene associated to Hirschsprung disease (HSCR) with differential contributions of its rare and common, coding and noncoding mutations to the multifactorial nature of this pathology. In the present study, we have performed a comprehensive study of our HSCR series evaluating the involvement of both RET rare variants (RVs) and common variants (CVs) in the context of the disease.

Partner Notification for Chlamydia Trachomatis Urogenital Infections: Eight Years of Patient Referral Experience in the Major Portuguese Sexually Transmitted Infections Clinic, 2000-07

International Journal of STD & AIDS. Oct, 2011  |  Pubmed ID: 21998173

The aim of this study was to perform a retrospective evaluation of the success of patient referral for partner notification of chlamydial infections to the major Portuguese sexually transmitted infection (STI) clinic. A total of 8277 patients were screened for Chlamydia trachomatis during 2000-2007, and 695 (8.4%) tested positive. The sexual partners of 34% of these index cases attended the clinic as contacts following partner referral. In univariate analysis, heterosexual men referred partners more frequently than men who have sex with men (MSM) and HIV-negative index cases referred partners four-fold more frequently than HIV-positives; however, these associations were non-significant after multivariate analysis (adjusted odds ratio [OR] 1.4, 95% confidence interval [CI] 0.7-2.6; adjusted OR 4.2, 95% CI 0.9-18.7, respectively). Index patients who reported lower numbers of partners referred more frequently, and this association remained significant after multivariate analysis. One-third of the referred individuals tested C. trachomatis-positive; the risk for infection was three-fold higher for referred partners from symptomatic index cases (P < 0.001, adjusted OR, 95% CI 1.8-6.3). In conclusion, the results of the present study concerning sexual behaviour, HIV status and clinical signs highlight the need for further evaluations that may shape future partner notification strategies in order to reduce the chlamydial disease burden.

Inoculation of Newborn Mice with Non-coding Regions of Foot-and-mouth Disease Virus RNA Can Induce a Rapid, Solid and Wide-range Protection Against Viral Infection

Antiviral Research. Dec, 2011  |  Pubmed ID: 22020303

We have recently described the ability of in vitro-transcribed RNAs, mimicking structural domains in the 5' and 3' non-coding regions (NCRs) of the foot-and-mouth disease virus (FMDV) genome, to trigger the innate immune response in porcine cultured cells and mice. In this work, the antiviral effect exerted in vivo by these small synthetic non-infectious RNA molecules was analyzed extensively. The susceptibility of transfected newborn Swiss mice to FMDV challenge was tested using a wide range of viral doses. The level of protection depended on the specific RNA inoculated and was dose-dependent. The RNA giving the best protection was the internal ribosome entry site (IRES), followed by the transcripts corresponding to the S fragment. The time course of resistance to FMDV of the RNA-transfected mice was studied. Our results show the efficacy of these RNAs to prevent viral infection as well as to contain ongoing FMDV infection in certain time intervals. Protection proved to be independent of the serotype of FMDV used for challenge. These results support the potential use of the FMDV NCR transcripts as both prophylactic and therapeutic molecules for new FMDV control strategies.

Differential Expression of CD300a/c on Human TH1 and TH17 Cells

BMC Immunology. 2011  |  Pubmed ID: 22046970

Human memory CD4+ T cells can be either CD300a/c+ or CD300a/c- and subsequent analyses showed that CD4+ effector memory T (T(EM)) cells are mostly CD300a/c+, whereas CD4+ central memory T (T(CM)) cells have similar frequencies of CD300a/c+ and CD300a/c- cells.

Mutation Screening of Multiple Genes in Spanish Patients with Autosomal Recessive Retinitis Pigmentosa by Targeted Resequencing

PloS One. 2011  |  Pubmed ID: 22164218

Retinitis Pigmentosa (RP) is a heterogeneous group of inherited retinal dystrophies characterised ultimately by the loss of photoreceptor cells. RP is the leading cause of visual loss in individuals younger than 60 years, with a prevalence of about 1 in 4000. The molecular genetic diagnosis of autosomal recessive RP (arRP) is challenging due to the large genetic and clinical heterogeneity. Traditional methods for sequencing arRP genes are often laborious and not easily available and a screening technique that enables the rapid detection of the genetic cause would be very helpful in the clinical practice. The goal of this study was to develop and apply microarray-based resequencing technology capable of detecting both known and novel mutations on a single high-throughput platform. Hence, the coding regions and exon/intron boundaries of 16 arRP genes were resequenced using microarrays in 102 Spanish patients with clinical diagnosis of arRP. All the detected variations were confirmed by direct sequencing and potential pathogenicity was assessed by functional predictions and frequency in controls. For validation purposes 4 positive controls for variants consisting of previously identified changes were hybridized on the array. As a result of the screening, we detected 44 variants, of which 15 are very likely pathogenic detected in 14 arRP families (14%). Finally, the design of this array can easily be transformed in an equivalent diagnostic system based on targeted enrichment followed by next generation sequencing.

The 10th Anniversary of the Junior Members and Affiliates of the European Academy of Allergy and Clinical Immunology

Pediatric Allergy and Immunology : Official Publication of the European Society of Pediatric Allergy and Immunology. Dec, 2011  |  Pubmed ID: 22122787

This year is the 10th anniversary of the European Academy of Allergy and Clinical Immunology (EAACI) Junior Members and Affiliates (JMAs). The aim of this review is to highlight the work and activities of EAACI JMAs. To this end, we have summarized all the initiatives taken by JMAs during the last 10 yr. EAACI JMAs are currently a group of over 2380 clinicians and scientists under the age of 35 yr, who support the continuous education of the Academy's younger members. For the past decade, JMAs enjoy a steadily increasing number of benefits such as free online access to the Academy's journals, the possibility to apply for Fellowships and the Mentorship Program, travel grants to attend scientific meetings, and many more. In addition, JMAs have been involved in task forces, cooperation schemes with other scientific bodies, organization of JMA focused sessions during EAACI meetings, and participation in the activities of EAACI communication platforms. EAACI JMA activities represent an ideal example of recruiting, training, and educating young scientists in order for them to thrive as future experts in their field. This model may serve as a prototype for other scientific communities, several of which have already adapted similar policies.

Maintenance of Previously Uncultured Freshwater Archaea from Anoxic Waters Under Laboratory Conditions

Antonie Van Leeuwenhoek. Feb, 2011  |  Pubmed ID: 20077005

Culture conditions for the maintenance of previously uncultured members of the Archaea thriving in anoxic water layers of stratified freshwater lakes are described. The proposed enrichment conditions, based on the use of defined medium composition and the maintenance of anoxia, have been proven effective for the maintenance of the archaeal community with virtually no changes over time for periods up to 6 months as revealed by a PCR-DGGE analysis. Phylotypes belonging to groups poorly represented in culture collections such as the Deep-Sea Hydrothermal Vent Euryarchaeota (DHVE) and the Miscellaneous Crenarchaeotic Group (MCG) were maintained and selectively enriched when compared to the correspondent indigenous planktonic archaeal community.

Somatic Mosaicism for Y120X Mutation in the MECP2 Gene Causes Atypical Rett Syndrome in a Male

Brain & Development. Aug, 2011  |  Pubmed ID: 20970936

Rett Syndrome (RS; MIM_312750) is a severe and progressive neurodevelopmental disorder affecting principally females. Mutations in X-Linked MECP2 gene (methyl CpG-binding protein 2; MIM_300005) have been reported as being the major cause of RS. Mutations in this gene have been described as cause of wide spectrum of neurological disorders and mental retardation in males. In some cases, mutations in MECP2 in males produce clinical picture similar to RS. Here we report the identification of the novel truncating mutation Y120X in a 4-year-old child with atypical RS phenotype. Chromosome analysis showed a normal karyotype, and blood DNA and tissue DNA analysis reveal a mosaic for the mutation. Patient's mother DNA analysis showed that this is a de novo mutation, that has never been described before in any female or male case of RS.

Novel Mutations at RET Ligand Genes Preventing Receptor Activation Are Associated to Hirschsprung's Disease

Journal of Molecular Medicine (Berlin, Germany). May, 2011  |  Pubmed ID: 21206993

Hirschsprung disease (HSCR) is a developmental disorder characterized by the absence of ganglion cells along variable lengths of the distal gastrointestinal tract. The major susceptibility gene for the disease is the RET proto-oncogene, which encodes a receptor tyrosine kinase activated by the glial cell-derived neurotrophic factor (GDNF) family ligands. We analyzed the coding sequence of GDNF, NTRN, and, for the first time, ARTN and PSPN in HSCR patients and detected several novel variants potentially involved in the pathogenesis of HSCR. In vitro functional analysis revealed that the variant R91C in PSPN would avoid the correct expression and secretion of the mature protein. Moreover, this study also highlighted the role of both this variant and F127L in NRTN in altering RET activation by a significant reduction in phosphorylation. To support the role of PSPN R91C in HSCR phenotype, enteric nervous system (ENS) progenitors were isolated from human postnatal gut tissues and expression of GFRα4, the main co-receptor for PSPN, was demonstrated. This suggests that not only GDNF and NRTN but also PSPN might promote survival of precursor cells during ENS development. In summary, we report for the first time the association of PSPN gene with HSCR and confirm the involvement of NRTN in the disease, with the identification of novel variants in those genes. Our results suggest that the biological consequence of the mutations NTRN F127L and PSPN R91C would be a reduction in the activation of RET-dependent signaling pathways, leading to a defect in the proliferation, migration, and/or differentiation process of neural crest cells within the developing gut and thus to the typical aganglionosis of the HSCR phenotype.

Evolutionary and Structural Features of the C2, V3 and C3 Envelope Regions Underlying the Differences in HIV-1 and HIV-2 Biology and Infection

PloS One. 2011  |  Pubmed ID: 21283793

Unlike in HIV-1 infection, the majority of HIV-2 patients produce broadly reactive neutralizing antibodies, control viral replication and survive as elite controllers. The identification of the molecular, structural and evolutionary footprints underlying these very distinct immunological and clinical outcomes may lead to the development of new strategies for the prevention and treatment of HIV infection.

[Portfolios: a Tool for the Training and Assessment of Residents in Dermatology, Part 1]

Actas Dermo-sifiliográficas. May, 2011  |  Pubmed ID: 21306691

The medical resident's portfolio is a collection of materials that show reflective learning in the context of clinical practice. A portfolio contains documents (such as case histories and questionnaires the resident has used), images, and video recordings that reveal that an individual has acquired the competencies needed for professional practice. A portfolio is an assessment tool that simultaneously supports learning and gives evidence for certifying competence. It encourages independent continuing professional development that is incremental and centered on answering questions about what one has learned, how it might be applied, what still needs to be learned, and what must be done to reach one's goal. Answering such questions provides evidence of competencies that have been acquired and what is still lacking, allowing the trainee to develop a plan for personal improvement and evaluate subsequent achievements. The first step in creating a portfolio is to list required skills and abilities, along with the actions that will allow the resident to acquire them during each year of residency training. The ultimate goal is to define the resident's professional competence. We describe a model on which to base a training and assessment portfolio for residents in dermatology.

DNA Immunization of Pigs with Foot-and-mouth Disease Virus Minigenes: from Partial Protection to Disease Exacerbation

Virus Research. Apr, 2011  |  Pubmed ID: 21315115

Despite several attempts to design new vaccines, there are as of yet no available alternatives to the conventional FMDV vaccines. Here, we present the divergent results obtained in pigs after immunization with two experimental DNA vaccines encoding one B and two T cell FMDV epitopes, either expressed alone (pCMV-BTT) or fused to a strong signal peptide (pCMV-spBTT). While all pigs vaccinated with pCMV-spBTT showed both a delay in the disease onset and reduced severity of signs and lesions after FMDV challenge, pigs immunized with pCMV-BTT showed an exacerbation of the disease and most of the pigs remained viremic at 10 days post-infection, the end-point of the experiment, thus opening concerns about FMDV-suboptimal immunization. Interestingly, only one of the four pigs vaccinated with pCMV-spBTT showed neutralizing antibodies before challenge, demonstrating that partial protection against FMDV could be afforded in the absence of preexisting neutralizing antibodies.

Accumulation of Distinct Prelamin A Variants in Human Diploid Fibroblasts Differentially Affects Cell Homeostasis

Experimental Cell Research. Feb, 2011  |  Pubmed ID: 20974128

Lamin A is a component of the nuclear lamina that plays a major role in the structural organization and function of the nucleus. Lamin A is synthesized as a prelamin A precursor which undergoes four sequential post-translational modifications to generate mature lamin A. Significantly, a large number of point mutations in the LMNA gene cause a range of distinct human disorders collectively known as laminopathies. The mechanisms by which mutations in lamin A affect cell function and cause disease are unclear. Interestingly, recent studies have suggested that alterations in the normal lamin A pathway can contribute to cellular dysfunction. Specifically, we and others have shown, at the cellular level, that in the absence of mutations or altered splicing events, increased expression of wild-type prelamin A results in a growth defective phenotype that resembles that of cells expressing the mutant form of lamin A, termed progerin, associated with Hutchinson-Gilford Progeria syndrome (HGPS). Remarkably, the phenotypes of cells expressing elevated levels of wild-type prelamin A can be reversed by either treatment with farnesyltransferase inhibitors or overexpression of ZMPSTE24, a critical prelamin A processing enzyme, suggesting that minor increases in the steady-state levels of one or more prelamin A intermediates is sufficient to induce cellular toxicity. Here, to investigate the molecular basis of the lamin A pathway toxicity, we characterized the phenotypic changes occurring in cells expressing distinct prelamin A variants mimicking specific prelamin A processing intermediates. This analysis demonstrates that distinct prelamin A variants differentially affect cell growth, nuclear membrane morphology, nuclear distribution of lamin A and the fundamental process of transcription. Expression of prelamin A variants that are constitutively farnesylated induced the formation of lamin A aggregates and dramatic changes in nuclear membrane morphology, which led to reduced levels of the basal transcription factor TATA-binding protein (TBP) and global transcription, and severely limited cell growth. Expression of a prelamin A variant that cannot be farnesylated, although did not appreciably influence cell growth, resulted in the formation of lamin A nucleoplasmic foci and caused, in a minor subpopulation of cells, changes in nuclear morphology that were accompanied by reduced levels of TBP and transcription. In contrast, expression of mature lamin A did not affect any of these parameters. These data demonstrate that accumulation of any partially processed prelamin A protein alters cellular homeostasis to some degree, even though the most dramatic effects are caused by variants with a permanently farnesylated carboxyl-terminal tail.

[Recommendations of Good Practices for the Genetic Diagnosis of Myotonic Dystrophy. Grupo AEGH/CIBERER]

Medicina Clínica. Mar, 2011  |  Pubmed ID: 20863536

Impairment of Emotional Expression Recognition in Schizophrenia: a Cuban Familial Association Study

Psychiatry Research. Jan, 2011  |  Pubmed ID: 20580837

It is well established that schizophrenia is associated with difficulties in recognizing facial emotional expressions, but few studies have reported the presence of this deficit among their unaffected relatives. This study attempts to add new evidence of familial association on an emotional expression processing test. The study evaluated the performance of 93 paranoid schizophrenia patients, 110 first-degree relatives of probands from multiplex schizophrenia families, and 109 nonpsychiatric controls on a facial emotional recognition test using a computer morphing technique to present the dynamic expressions. The task entailed the recognition of a set of facial expressions depicting the six basic emotions presented in 21 successive frames of increasing intensity. The findings indicated that schizophrenia patients were consistently impaired for the recognition of the six basic facial expressions. In contrast, their unaffected relatives showed a selective impairment for the recognition of disgust and fearful expressions. Familial association of selective facial emotional expressions processing deficit may further implicate promising new endophenotypes that can advance the understanding of affective deficits in schizophrenia.

Biofouling and Biodeterioration in Materials Stored at the Historical Archive of the Museum of La Plata, Argentine and at the National Archive of the Republic of Cuba

Colloids and Surfaces. B, Biointerfaces. Jul, 2011  |  Pubmed ID: 21439796

The aims of this paper were to study the biofouling and biodeterioration of photos and maps stored at Historical Archive of the Museum of La Plata (HAMP), Argentine, and two repositories of the National Archive of Cuba Republic (NARC) and to carry out the physiological characterization of the isolated fungi and bacteria. The role of the environmental microbiota in the biofouling formation was also studied. Microbial assemblages in the air were sampled by sedimentation technique while those on documents were sampled by swabbering. Biofilm formation and biofouling were monitored by scanning electron microscope (SEM). Large microbial assemblages were found at NARC archives with the prevalence of genera Aspergillus, Cladosporium and Penicillium, whereas at HAMP these values were lower, Penicillium was the only fungal genus detected. Most of the fungi degraded cellulose and produced pigments and acids, and all of the isolated bacteria had proteolytic and/or cellulolytic activity. In all cases, a higher concentration of viable bacteria than of fungi was isolated from documents. These results correlated with bacterial values detected in air at NARC repositories. However, this correlation cannot be observed at HAMP where Aspergillus, Penicillium and Talaromyces helicus (teleomorph of Penicillium) were isolated. It is the first time that the last genus is reported in documents.

The Impact of Anthropometric Patient-phantom Matching on Organ Dose: a Hybrid Phantom Study for Fluoroscopy Guided Interventions

Medical Physics. Feb, 2011  |  Pubmed ID: 21452738

To investigate the benefits and limitations of patient-phantom matching for determining organ dose during fluoroscopy guided interventions.

Pathologic Complete Responses After Chemotherapy Plus Figitumumab in Stage IV Non-small-cell Lung Cancer

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Mar, 2011  |  Pubmed ID: 21149666

Impact of Land Use on Urban Mobility Patterns, Emissions and Air Quality in a Portuguese Medium-sized City

The Science of the Total Environment. Feb, 2011  |  Pubmed ID: 21216441

The main objective of this work was to evaluate the impact of urban development trends in mobility patterns of a medium sized Portuguese city and air quality consequences, using a sequential modeling process, comprising i) land use and transportation, TRANUS model; ii) road traffic air pollutants emissions, TREM model and; iii) air quality, TAPM model. This integrated methodology was applied to a medium sized Portuguese city. In order to evaluate the implementation of the methodology, a preliminary study was performed, which consisted on the comparison of modeled mobility patterns and CO and PM(10) concentrations with measured data used in the definition of the current scenario. The comparison between modeled and monitored mobility patterns at the morning peak hour for a weekday showed an RMSE of 31%. Regarding CO concentrations, an underestimation of the modeled results was observed. Nevertheless, the modeled PM(10) concentrations were consistent with the monitored data. Overall, the results showed a reasonable consistency of the modeled data, which allowed the use of the integrated modeling system for the study scenarios. The future scenarios consisted on the definition of different mobility patterns and vehicle technology characteristics, according to two main developing trends: (1) "car pooling" scenario, which imposes a mean occupancy rate of 3 passengers by vehicle and (2) the "Euro 6" scenario, which establishes that all vehicles accomplish at least the Euro 6 standard technology. Reductions of 54% and 83% for CO, 44% and 95% for PM(10), 44% and 87% for VOC and 44% and 79% for NO(x) emissions were observed in scenarios 1 and 2, respectively. Concerning air quality, a reduction of about 100 μg m(-3) of CO annual average concentration was observed in both scenarios. The results of PM(10) annual concentrations showed a reduction of 1.35 μg m(-3) and 2.7 μg m(-3) for scenarios 1 and 2 respectively.

Age-related Sensitization to P-phenylenediamine

Contact Dermatitis. Mar, 2011  |  Pubmed ID: 21272028

Clinical and Epidemiological Characteristics of Patients with Influenza A (H1N1) 2009 Attended to at the Emergency Room of a Children's Hospital

European Journal of Pediatrics. Mar, 2011  |  Pubmed ID: 21274563

In June 2009, the first influenza pandemic of the twenty-first century, due to the swine origin influenza A (H1N1) 2009 virus, was declared. This study aimed to describe the epidemiological and clinical features, complications, lethality and risk factors for hospital admission of microbiologically confirmed cases of influenza A (H1N1) 2009 infection seen at the emergency department of a children's hospital. All cases of children with influenza A (H1N1) 2009 viral infection, confirmed microbiologically by real-time reverse transcription polymerase chain reactions and treated in the emergency room between July and December 2009, were prospectively included. Patients were compared according to admission requirement to study variables associated with the risk of hospitalisation. Oseltamivir was the antiviral used for the treatment and its safety was analysed. Four hundred and twelve patients with influenza A (H1N1) 2009 infection were included. The most frequent symptoms were: fever (96%), cough (95%) and coryza (90%). Eighty-five patients (20.6%) were admitted: three to the paediatric intensive care unit and two died. Hospitalised children were younger than those not admitted (median age 5 vs 8 years; p = 0.001). Age under 1 year (OR 6.01; CI 95% 2.77-13.05), pneumonia (OR 7.99; CI 95% 3.50-18.22) and haemoglobinopathy or underlying blood disorders (OR 5.99; CI 95% 1.32-27.30) were statistically significant risk factors for admission. No differences were observed regarding onset of antiviral treatment among admitted and non-admitted patients. Treatment with oseltamivir was well tolerated. In conclusion, the incidence of severe cases and lethality of influenza A (H1N1) 2009 infection were low in our setting, even in a population with risk factors for developing complications.

Current Strategies for Subunit and Genetic Viral Veterinary Vaccine Development

Virus Research. Apr, 2011  |  Pubmed ID: 21316403

Developing vaccines for livestock provides researchers with the opportunity to perform efficacy testing in the natural hosts. This enables the evaluation of different strategies, including definition of effective antigens or antigen combinations, and improvement in delivery systems for target antigens so that protective immune responses can be modulated or potentiated. An impressive amount of knowledge has been generated in recent years on vaccine strategies and consequently a wide variety of antigen delivery systems is now available for vaccine research. This paper reviews several antigen production and delivery strategies other than those based on the use of live viral vectors. Genetic and protein subunit vaccines as well as alternative production systems are considered in this review.

CD300a is Expressed on Human B Cells, Modulates BCR-mediated Signaling, and Its Expression is Down-regulated in HIV Infection

Blood. Jun, 2011  |  Pubmed ID: 21482706

The immunomodulatory receptor CD300a is expressed on human B cells. Naive B cells express very low levels of this receptor, whereas memory B cells and plasmablasts/cells express variable levels of CD300a. Germinal center B cells are negative for CD300a expression. Stimulation of naive B cells via B-cell receptor (BCR) and Toll-like receptor 9, along with T-cell help, failed to up-regulate CD300a cell surface expression despite the increased expression of the memory marker CD27 and the down-regulation of CD305. However, Toll-like receptor 9 stimulation alone significantly increased CD300a expression on memory B cells, whereas interleukin-4 and transforming growth factor-β1 act as negative regulators of CD300a expression on memory B cells. Coligation of BCR and CD300a inhibits Ca(2+) mobilization and nuclear factor of activated T cell transcriptional activity evoked by BCR ligation alone. Suppression of CD300a expression in primary B cells with siRNA resulted in increased BCR-mediated proliferation, thereby confirming the inhibitory capacity of CD300a. Finally, we show that CD300a expression levels are significantly down-regulated in the circulating B cells of HIV-infected patients. Altogether, these data demonstrate a novel mechanism for suppressing the activity of B cells and suggest a potential role for CD300a in the B-cell dysfunction observed in HIV-induced immunodeficiency.

Active Bacteria and Archaea Cells Fixing Bicarbonate in the Dark Along the Water Column of a Stratified Eutrophic Lagoon

FEMS Microbiology Ecology. Aug, 2011  |  Pubmed ID: 21517915

We studied the carbon dioxide fixation activity in a stratified hypereutrophic karstic lagoon using a combination of fingerprinting techniques targeting bacterial and archaeal 16S rRNA genes, functional gene cloning [the acetyl-CoA carboxylase (accC)], and isotopic labelling ((14)C-bicarbonate) coupled to single-cell analyses [microautoradiography combined with catalyzed reported deposition-FISH (MAR-CARD-FISH)]. The microbial planktonic community was dominated by bacteria with maximal abundances of archaea just below the oxic/anoxic transition zone (7% of total cells). In situ incubations with radiolabelled bicarbonate showed maximal photoassimilation activity in the oxic epilimnion, whereas dark CO(2) fixation was consistently observed throughout the water column, with a maximum at the oxic/anoxic interface (8.6 mg C m(-3) h(-1)). The contributions of light and dark carbon fixation activities in the whole water column were 69% and 31% of the total C incorporated, respectively. MAR-CARD-FISH incubations corroborated these results and revealed that the highest fraction of bacterial and archaeal cells actively uptaking bicarbonate in the light was found at the surface. The bacterial community was mainly composed of green sulfur bacteria (Chlorobi) and members of the Betaproteobacteria and the Bacteroidetes. The archaeal assemblage was composed of phylotypes of the Miscellaneous Crenarchaeotic Group and a few methanogens. Clone libraries of the accC gene showed an absolute dominance of bacterial carboxylases. Our results suggest that the dark carbon fixation activity measured was mainly related to CO(2) incorporation by heterotrophs rather than to the activity of true chemoautotrophs.

Copy-number Variations in EYS: a Significant Event in the Appearance of ArRP

Investigative Ophthalmology & Visual Science. Jul, 2011  |  Pubmed ID: 21519034

Autosomal recessive retinitis pigmentosa (arRP) has recently been associated with mutations in a novel gene, EYS, which is a major gene for this disease. All published mutations so far are based on conventional PCR and are not adequate to identify midsized DNA rearrangements. This study was conducted to establish the prevalence of copy-number variations (CNVs) in the EYS gene in a cohort of arRP patients, including individuals in whom only one pathogenic change was detected by PCR-based sequencing.

RNA Structural Domains in Noncoding Regions of the Foot-and-mouth Disease Virus Genome Trigger Innate Immunity in Porcine Cells and Mice

Journal of Virology. Jul, 2011  |  Pubmed ID: 21525336

The induction of type I interferons (alpha/beta interferon [IFN-α/β]) in response to viral infection is a crucial step leading to the antiviral state in the host. Viruses produce double-stranded RNA (dsDNA) during their replication cycle that is sensed as nonself by host cells through different receptors. A signaling cascade then is activated to block viral replication and spread. Foot-and-mouth disease virus (FMDV) is a picornavirus that is highly sensitive to IFN, and it causes one of the world's most important animal diseases. In this study, we showed the ability of structural domains predicted to enclose stable dsRNA regions in the 5'- and 3'-noncoding regions (NCRs) of the FMDV genome to trigger an IFN-α/β response in porcine kidney cultured cells and newborn mice. These RNAs, generated by in vitro transcription, were able to stimulate IFN-β transcription and induce an antiviral state in SK-6 cells. The induction levels elicited by the different NCR RNAs were compared. Among them, the 3'NCR was identified as a potent IFN activator, and the features in this region involved in signaling have been analyzed. To address whether the FMDV NCR transcripts were able to trigger the innate immune response in vivo, Swiss suckling mice were inoculated intraperitoneally with the RNAs. All transcripts induced the innate response in transfected animals, measured as IFN-α/β protein levels, antiviral activity in sera, and reduced susceptibility to FMDV infection. Our work provides new insight into innate responses against FMDV and identifies these small noninfectious RNA molecules as potential adjuvants for vaccine improvement and antiviral strategies against picornaviruses.

[Indications for Referral to a Skin Allergy Unit]

Actas Dermo-sifiliográficas. Jul-Aug, 2011  |  Pubmed ID: 21529723

Contact dermatitis is one of the most common reasons for consultation in dermatology. However, general dermatologists do not always appreciate the importance of patch testing. These tests should ideally be performed in specialist skin allergy units, most importantly in cases suggestive of contact dermatitis, severe acute dermatitis, chronic persistent dermatitis, and dermatitis affecting the eyelids, genital region or adjacent to venous ulcers. Eczematous changes in pre-existing skin lesions or lesions at atypical sites in patients diagnosed with atopic eczema should also be investigated. Finally, cases diagnosed as occupational dermatitis can be best managed by the workers' health insurance scheme.

[Portfolios: a Tool for the Training and Assessment of Residents in Dermatology, Part 2]

Actas Dermo-sifiliográficas. Jun, 2011  |  Pubmed ID: 21531360

A portfolio is a collection of material documenting reflection about practice. It contains documents (eg, case histories and questionnaires the resident has used), images, and video recordings that reveal that an individual has acquired the competencies needed for professional practice. This assessment tool simultaneously supports learning and provides evidence for certifying competence. The adoption of portfolio use by a dermatology department requires the support of both the training supervisor and the chief of department. The learning objectives defined by the National Board for Medical-Surgical Dermatology and Venereology must be taken into consideration so that ways to assess each objective can be included; this approach supports holistic ongoing education as well as the certification of competencies the resident finally achieves. Use of portfolios in medical residency training can improve on current assessment methods, which we believe lack precision. We propose that portfolios gradually begin to replace the resident's training log. We are currently developing an online software application that will facilitate the use of portfolios.

Correlation Between Lesion Site and Concentration of Dimethyl Fumarate in Different Parts of Shoes in Patients with Contact Dermatitis Caused by Dimethyl Fumarate in Footwear

Contact Dermatitis. Aug, 2011  |  Pubmed ID: 21545603

Dimethyl fumarate (DMF) has been identified as being responsible for an outbreak of shoe contact dermatitis in Europe. All reported cases to date have involved the dorsa of the toes and the dorsa of the feet, sometimes in association with other areas.

Resolution of IgE-mediated Fish Allergy

Allergologia Et Immunopathologia. Jun, 2011  |  Pubmed ID: 21640463

Hospital Resource Utilization and Treatment Cost of Skeletal-related Events in Patients with Metastatic Breast or Prostate Cancer: Estimation for the Portuguese National Health System

Value in Health : the Journal of the International Society for Pharmacoeconomics and Outcomes Research. Jun, 2011  |  Pubmed ID: 21669375

Skeletal-related events (SREs) occur frequently in patients with bone metastases as a result of breast (BC) and prostate (PC) cancers. They increase both morbidity and mortality and lead to extensive health-care resource utilization.

Enhancing DNA Immunization by Targeting ASFV Antigens to SLA-II Bearing Cells

Vaccine. Jul, 2011  |  Pubmed ID: 21679736

One of the main criticisms to DNA vaccines is the poor immunogenicity that they confer on occasions, at least in large animals. Confirming this theory, immunization with plasmid DNA encoding two African swine fever virus genes in frame (pCMV-PQ), failed in inducing detectable immune responses in pigs, while it was successful in mice. Aiming to improve the immune responses induced in swine, a new plasmid was constructed, encoding the viral genes fused in frame with a single chain variable fragment of an antibody specific for a swine leukocyte antigen II (pCMV-APCH1PQ). Our results clearly demonstrate that targeting antigens to antigen professional cells exponentially enhanced the immune response induced in pigs, albeit that the DNA vaccine was not able to confer protection against lethal viral challenge. Indeed, a viremia exacerbation was observed in each of the pigs that received the pCMV-APCH1PQ plasmid, this correlating with the presence of non-neutralizing antibodies and antigen-specific SLA II-restricted T-cells. The implications of our discoveries for the development of future vaccines against African swine fever virus and other swine pathogens are discussed.

Patch Testing in Children with Hand Eczema. A 5-year Multicentre Study in Spain

Contact Dermatitis. Oct, 2011  |  Pubmed ID: 21702757

Hand eczema is common in children, but affected children are seldom patch tested. Relatively few studies have assessed patch testing in the paediatric population, and none has specifically evaluated its use in hand eczema in children.

Paradoxical Response During Antituberculosis Therapy in a Patient with Tuberculosis Verrucosa Cutis

Journal of the American Academy of Dermatology. Aug, 2011  |  Pubmed ID: 21763552

The Spectrum of Pneumococcal Empyema in Adults in the Early 21st Century

Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America. Aug, 2011  |  Pubmed ID: 21765073

Increased rates of empyema have been reported in children after the introduction of the pneumococcal conjugate vaccine (PCV7). Our objective was to describe the risk factors for pneumococcal empyema in adults and to analyze the differences in the incidence, disease characteristics, and serotype distribution between the pre- and post-PCV7 eras.

Patch Testing with Formaldehyde and Formaldehyde-releasers: Multicentre Study in Spain (2005-2009)

Contact Dermatitis. Nov, 2011  |  Pubmed ID: 21767276

Formaldehyde and formaldehyde-releasers are common causes of allergic contact dermatitis.

Usefulness of Endoscopic Ultrasonography in the Clinical Suspicion of Biliary Disease

Revista Española De Enfermedades Digestivas : Organo Oficial De La Sociedad Española De Patología Digestiva. Jul, 2011  |  Pubmed ID: 21770679

endoscopic ultrasound (EUS) is a very sensitive and specific technique for the diagnosis of biliary diseases. This procedure has proven its usefulness in cases of high suspicion of biliary disease (history of gallstones and dilatation of the intrahepatic and/or extrahepatic bile ducts). We know less about its usefulness in cases of low suspicion of biliary pathology.The aim of this study was to assess the diagnostic accuracy of EUS in patients with low suspicion of biliary disease (patients with dilatation of the biliary tract were excluded).

Tissue Distribution of Red Spotted Grouper Nervous Necrosis Virus (RGNNV) Genome in Experimentally Infected Juvenile European Seabass (Dicentrarchus Labrax)

Veterinary Microbiology. Dec, 2011  |  Pubmed ID: 21783331

The distribution of viral genome in the tissues of juvenile European seabass (Dicentrarchus labrax) during the course of a Red Spotted Grouper Nervous Necrosis Virus (RGNNV) infection has not yet been described. The present study addresses this and indicates which target organs may be involved in viral replication. This information should enable more accurate detection of virus in asymptomatic carriers, and in turn help to control the spread of the disease. The aim of this study was to examine the pattern of expression of viral genomic segments RNA1 and RNA2, using two absolute real-time PCRs (RT-qPCR), over the course of a RGNNV infection after administering the virus by intramuscular injection. In situ hybridization was also used to locate the RNA2 viral segment in different organs throughout the infection. The experimental challenge provoked an acute form of viral nervous necrosis (VNN), with a resulting cumulative mortality of 37%. The RT-qPCRs designed allowed the detection of both genomic segments in all the organs tested (nervous and non-nervous tissues) at all sampling times examined. The highest viral RNA copy number was found in eyes, although viral replication appeared to begin in the brain. Viral replication was also recorded in pooled internal organs and in caudal fin. However, the increase in the viral RNA copy number in these organs did not result in an increased viral titre, which may indicate that a productive infection does not take place in non-nervous tissues, possibly due to a failure in a viral post-replication step.

Complex Regulation of Human NKG2D-DAP10 Cell Surface Expression: Opposing Roles of the γc Cytokines and TGF-β1

Blood. Sep, 2011  |  Pubmed ID: 21816829

Natural killer (NK) cells help protect the host against viral infections and tumors. NKG2D is a vital activating receptor, also expressed on subsets of T cells, whose ligands are up-regulated by cells in stress. Ligation of NKG2D leads to phosphorylation of the associated DAP10 adaptor protein, thereby activating immune cells. Understanding how the expression of NKG2D-DAP10 is regulated has implications for immunotherapy. We show that IL-2 and TGF-β1 oppositely regulate NKG2D-DAP10 expression by NK cells. IL-2 stimulation increases NKG2D surface expression despite a decrease in NKG2D mRNA levels. Stimulation with IL-2 results in a small increase of DAP10 mRNA and a large up-regulation of DAP10 protein synthesis, indicating that IL-2-mediated effects are mostly posttranscriptional. Newly synthesized DAP10 undergoes glycosylation that is required for DAP10 association with NKG2D and stabilization of NKG2D expression. TGF-β1 has an opposite and dominant effect to IL-2. TGF-β1 treatment decreases DAP10, as its presence inhibits the association of RNA polymerase II with the DAP10 promoter, but not NKG2D mRNA levels. This leads to the down-regulation of DAP10 expression and, as a consequence, NKG2D protein as well. Finally, we show that other γ(c) cytokines act similarly to IL-2 in up-regulating DAP10 expression and NKG2D-DAP10 surface expression.

A DNA Vaccine Encoding Foot-and-mouth Disease Virus B and T-cell Epitopes Targeted to Class II Swine Leukocyte Antigens Protects Pigs Against Viral Challenge

Antiviral Research. Nov, 2011  |  Pubmed ID: 21820470

Development of efficient and safer vaccines against foot-and-mouth disease virus (FMDV) is a must. Previous results obtained in our laboratory have demonstrated that DNA vaccines encoding B and T cell epitopes from type C FMDV, efficiently controlled virus replication in mice, while they did not protect against FMDV challenge in pigs, one of the FMDV natural hosts. The main finding of this work is the ability to improve the protection afforded in swine using a new DNA-vaccine prototype (pCMV-APCH1BTT), encoding FMDV B and T-cell epitopes fused to the single-chain variable fragment of the 1F12 mouse monoclonal antibody that recognizes Class-II Swine Leukocyte antigens. Half of the DNA-immunized pigs were fully protected upon viral challenge, while the remaining animals were partially protected, showing a delayed, shorter and milder disease than control pigs. Full protection in a given vaccinated-pig correlated with the induction of specific IFNγ-secreting T-cells, detectable prior to FMDV-challenge, together with a rapid development of neutralizing antibodies after viral challenge, pointing towards the relevance that both arms of the immune response can play in protection. Our results open new avenues for developing future FMDV subunit vaccines.

Deep-vein Thrombosis: a United States Cost Model for a Preventable and Costly Adverse Event

Thrombosis and Haemostasis. Sep, 2011  |  Pubmed ID: 21833446

Preventable venous thromboembolism (VTE) and "appropriate" type, dose, and duration of prophylaxis are emerging concepts. Contemporary definitions by key quality organisations, including the World Health Organization, have shifted towards "preventable" VTE being considered an adverse event or adverse drug event. A decision tree and cost model were developed to estimate the United States health care costs for total deep-vein thrombosis (DVT), total hospital-acquired DVT, and total "preventable" DVT. Annual cost ranges were obtained in 2010 US dollars for total ($7.5 to $39.5 billion), hospital-acquired ($5 to $26.5 billion), and preventable ($2.5 to $19.5 billion) DVT costs. When the sensitivity analysis was applied--taking into consideration higher incidence rates and costs - annual US total, hospital-acquired, and "preventable" DVT costs ranged from $9.8 to $52 billion, $6.8 to $36 billion, and $3.4 to $27 billion, respectively.

[Ablation of Premature Ventricular Contractions Originating in the Right Ventricular Outflow Tract Using Non-contact Mapping]

Revista Española De Cardiología. Dec, 2011  |  Pubmed ID: 21835534

Premature ventricular contractions originating in the right ventricular outflow tract may respond poorly to pharmacological treatment, and ablation using conventional fluoroscopically-guided systems may be complicated by the difficulty in inducing arrhythmias. We describe the use of a non-contact mapping system to ablate difficult-to-induce premature ventricular contractions originating in the right ventricular outflow tract. Five premature ventricular contractions sites in the right ventricular outflow tract were ablated in a prospective series of 4 patients. Patients had a poor quality of life and had not responded to antiarrhythmic drugs. A mean of 3 radiofrequency pulses per site was applied and mean application time was 113±15s. We achieved a 100% acute success rate and there were no complications. Patients were asymptomatic without drug therapy after a mean of 30±16 months of follow-up. The noncontact mapping system is highly effective in eliminating difficult to induce, isolated premature ventricular contractions.

Expression of PROKR1 and PROKR2 in Human Enteric Neural Precursor Cells and Identification of Sequence Variants Suggest a Role in HSCR

PloS One. 2011  |  Pubmed ID: 21858136

The enteric nervous system (ENS) is entirely derived from neural crest and its normal development is regulated by specific molecular pathways. Failure in complete ENS formation results in aganglionic gut conditions such as Hirschsprung's disease (HSCR). Recently, PROKR1 expression has been demonstrated in mouse enteric neural crest derived cells and Prok-1 was shown to work coordinately with GDNF in the development of the ENS.

Cutting Edge: Mouse CD300f (CMRF-35-like Molecule-1) Recognizes Outer Membrane-exposed Phosphatidylserine and Can Promote Phagocytosis

Journal of Immunology (Baltimore, Md. : 1950). Oct, 2011  |  Pubmed ID: 21865548

Reportedly, CD300f negatively regulates interactions between dendritic and T cells and acts as an anti-inflammatory molecule in a multiple sclerosis mouse model. We found that a CD300f/Fc chimeric protein specifically binds to apoptotic/dead splenocytes and to apoptotic cells from starved or irradiated lymphocytic cell lines, an observation extended to insect cells. CD300f also binds PMA/ionomycin-activated splenocytes and Ag-stimulated T cells, an interaction inhibited by Annexin V. By ELISA, cosedimentation, and surface plasmon resonance using phospholipid-containing liposomes, we show that CD300f preferentially binds phosphatidylserine and requires a metal ion. Exogenous expression of CD300f in cell lines results in enhanced phagocytosis of apoptotic cells. We conclude that expression of CD300f conveys additional capacity to recognize phosphatidylserine to myeloid cells. The result of this recognition may vary with the overall qualitative and quantitative receptor content, as well as signaling capacity of the expressing effector cell, but enhanced phagocytosis is one measurable outcome.

Differential Antiviral Activity of Mx1, Mx2 and Mx3 Proteins from Gilthead Seabream (Sparus Aurata) Against Infectious Pancreatic Necrosis Virus (IPNV)

Molecular Immunology. Oct, 2011  |  Pubmed ID: 21872932

Mx proteins are crucial effectors of the innate antiviral response mediated by the interferon type I signalling pathway. Recently, three Mx proteins, named SauMx1, SauMx2 and SauMx3, corresponding to three different genes, have been identified in the cultured marine species gilthead seabream (Sparus aurata). In this study, the three SauMx cDNAs were cloned into expression vectors and used for transfection of CHSE-214 cells. Monoclonal cell populations stably expressing each recombinant protein have been obtained and characterized. The protection conferred by each recombinant SauMx against Infectious Pancreatic Necrosis Virus (IPNV) infection has been in vitro evaluated, having found clear differences among them. According to the cytopathic effects and the virus yield reduction assays, only cells expressing SauMx2 and SauMx3 showed significant resistance to IPNV infection. Otherwise, quantitative RT real-time PCR assays suggested that each SauMx protein has a different target during the viral inhibition process. The differences observed among the three SauMx proteins are discussed in terms of their differential mechanism of action and antiviral specificity, suggesting, as a whole, to play a synergistic activity in the protection of gilthead seabream against IPNV.

Effect of Fondaparinux Prophylaxis on Anti-factor Xa Concentrations in Patients with Morbid Obesity

American Journal of Health-system Pharmacy : AJHP : Official Journal of the American Society of Health-System Pharmacists. Sep, 2011  |  Pubmed ID: 21880887

Anti-factor Xa values in morbidly obese patients receiving standard doses of fondaparinux sodium for the prevention of venous thromboembolism (VTE) were analyzed in a retrospective chart evaluation.

Effectiveness of Antibiotic-lock Therapy for Long-term Catheter-related Bacteremia Due to Gram-negative Bacilli: a Prospective Observational Study

Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America. Nov, 2011  |  Pubmed ID: 21890750

A prospective observational study evaluated the effectiveness of combining antibiotic-lock therapy and systemic antibiotics for Gram-negative bacilli long-term catheter-related bacteremia. In 46 uncomplicated episodes, the most frequently isolated microorganisms were Pseudomonas aeruginosa (15), Enterobacter cloacae (12), Escherichia coli (10), and Klebsiella spp. (8). Cure was achieved in 95% of cases.

Genetic Diversity and Drug Resistance Profiles in HIV Type 1- and HIV Type 2-Infected Patients from Cape Verde Islands

AIDS Research and Human Retroviruses. Sep, 2011  |  Pubmed ID: 21902592

Abstract Our aim was to characterize for the first time the genetic diversity of HIV in Cape Verde Islands as well as the drug resistance profiles in treated and untreated patients. Blood specimens were collected from 41 HIV-1 and 14 HIV-2 patients living in Santiago Island. Half of the patients were on antiretroviral treatment (ART). Pol and env gene sequences were obtained using in-house methods. Phylogenetic analysis was used for viral subtyping and the Stanford Algorithm was used for resistance genotyping. For HIV-1, the amplification of pol and env was possible in 27 patients (66%). HIV-1 patients were infected with subtypes G (13, 48%), B (2, 7%), F1 (2, 7%), and CRF02_AG (2, 7%), and complex recombinant forms including a new C/G variant (n=8, 30%). Drug resistance mutations were detected in the PR and RT of three (10%) treated patients. M41L and K103N transmitted drug resistance mutations were found in 2 of 17 (12%) untreated patients. All 14 HIV-2 isolates belonged to group A. The origin of 12 strains was impossible to determine whereas two strains were closely related to the historic ROD strain. In conclusion, in Cape Verde there is a long-standing HIV-2 epidemic rooted in ROD-like strains and a more recent epidemic of unknown origin. The HIV-1 epidemic is caused by multiple subtypes and complex recombinant forms. Drug resistance HIV-1 strains are present at moderate levels in both treated and untreated patients. Close surveillance in these two populations is crucial to prevent further transmission of drug-resistant strains.

[Indications for Referral to a Skin Allergy Unit]

Actas Dermo-sifiliográficas. Jul, 2011  |  Pubmed ID: 21944772

Contact dermatitis is one of the most common reasons for consultation in dermatology. However, general dermatologists do not always appreciate the importance of patch testing. These tests should ideally be performed in specialist skin allergy units, most importantly in cases suggestive of contact dermatitis, severe acute dermatitis, chronic persistent dermatitis, and dermatitis affecting the eyelids, genital region or adjacent to venous ulcers. Eczematous changes in pre-existing skin lesions or lesions at atypical sites in patients diagnosed with atopic eczema should also be investigated. Finally, cases diagnosed as occupational dermatitis can be best managed by the workers' health insurance scheme.

Obstetric Outcomes After Treatment for Cervical Intraepithelial Neoplasia: Six Years of Experience

Journal of Lower Genital Tract Disease. Oct, 2011  |  Pubmed ID: 21959571

: The purpose of this study was to assess the effect of cervical conization on subsequent pregnancy and delivery outcomes.

Origin and Epidemiological History of HIV-1 CRF14_BG

PloS One. 2011  |  Pubmed ID: 21969855

CRF14_BG isolates, originally found in Spain, are characterized by CXCR4 tropism and rapid disease progression. This study aimed to identify the origin of CRF14_BG and reconstruct its epidemiological history based on new isolates from Portugal.

Quantification of P-phenylenediamine and 2-hydroxy-1,4-naphthoquinone in Henna Tattoos

Contact Dermatitis. Jan, 2012  |  Pubmed ID: 21972791

Very few studies are available in which the components of henna products used by tattoo artists have been analysed.

Unexpectedly High Frequency of European Parentage in Venezuelan Patients with Chronic Lymphocytic Leukemia

Leukemia & Lymphoma. Feb, 2012  |  Pubmed ID: 21867460

There is insufficient information on the characteristics of chronic lymphocytic leukemia (CLL) in Latin American patients. Immunoglobulin variable-region heavy-chain (IGVH) gene usage and mutation status and prognostic factors were investigated in patients resident in Venezuela. The most frequently used IGVH family genes were: VH3 > VH1 > VH4 > VH5, with a high incidence of IGVH1.69 and IGVH3.21 genes, and 55.2% of IGVH genes were mutated. Analysis of HCDR3 (third complementarity-determining region of the heavy chain) revealed that 24% of Venezuelan HCDR3s belonged to a CLL stereotyped HCDR3. Results for prognostic factors were similar to those reported previously for Caucasian populations. Interestingly, we found an over-representation of people of European extraction among Venezuelan patients with CLL, suggesting the possibility of a higher frequency of susceptibility genes for CLL in Europeans in comparison with Latin American mestizos.

Airways Changes Related to Air Pollution Exposure in Wheezing Children

The European Respiratory Journal : Official Journal of the European Society for Clinical Respiratory Physiology. Feb, 2012  |  Pubmed ID: 21719492

In this study, we aimed to evaluate the relationship between individual total exposure to air pollution and airway changes in a group of 51 wheezing children. Respiratory status was assessed four times (January 2006, June 2006, January 2007 and June 2007) during a 1-week period through a standardised questionnaire, spirometry, exhaled nitric oxide fraction and pH in exhaled breath condensate (EBC). Concentrations of particles with a 50% cut-off aerodynamic diameter of 10 µm (PM(10)), O(3), NO(2) and volatile organic compounds were estimated through direct measurements with an ad hoc device or air pollution modelling in the children's schools and at their homes in the same 4 weeks of the study. For each child, total exposure to the different air pollutants was estimated as a function of pollutant concentrations and daily activity patterns. Increasing total exposure to PM(10), NO(2), benzene, toluene and ethylbenzene was significantly associated with a decrease of forced expiratory volume in 1 s (FEV(1)) and with an increase of change in FEV(1). Increasing exposure to NO(2) and benzene was also related to a significant decrease of FEV(1)/forced vital capacity. Increasing exposure to PM(10), NO(2), benzene and ethylbenzene was associated with acidity of EBC. This study suggests an association in wheezing children between airway changes and total exposure to air pollutants, as estimated by taking into account the concentration in the various microenvironments attended by the children.

[Urinary Tract Infections and Their Prevention]

Actas Urologicas Españolas. Jan, 2012  |  Pubmed ID: 21757260

This article reviews diverse aspects of the prevention of urinary tract infections, including confirmation of the diagnosis, application of hygiene and dietary measures, antibacterial prophylaxis (preferably consisting of a single nocturnal oral dose per day of an antibiotic or drug with high urinary excretion and good tolerance), and administration of vaccines made with Escherichia coli and other Gram-negative bacilli, consisting of immunostimulating fractions of E. coli strains or E. coli type-1 fimbriae administered through the parenteral or oral route.

Pediatric Dermatology Consultations: a Retrospective Analysis of Inpatient Consultations Referred to the Dermatology Service

Pediatric Dermatology. Jan-Feb, 2012  |  Pubmed ID: 21453306

To analyze the care provided by consultant dermatologists to hospitalized pediatric patients, we retrospectively studied the records of inpatient pediatric consultation requests received by a hospital dermatology department between 2000 and 2009. The diagnoses were recorded according to the International Classification of Diseases, Ninth Revision (ICD-9). In a high percentage of cases, the problem was addressed in a single visit, and few additional tests were ordered. The most common diagnostic groups were diseases of the skin and subcutaneous tissue (ICD-9: 680-709). The most frequent specific diagnosis was atopic dermatitis (ICD-9: 691.8). Our series shows that the role of dermatologists as consultants for pediatric inpatients is similar to their role in outpatient consultations.

Usefulness of Perfusion CT to Assess Response to Neoadjuvant Combined Chemoradiotherapy in Patients with Locally Advanced Rectal Cancer

Academic Radiology. Feb, 2012  |  Pubmed ID: 22130088

To prospectively evaluate perfusion computed tomography (CT) for assessment of changes in tumor vascularity after chemoradiation therapy (CRT) in locally advanced rectal cancer and to analyze the correlation between baseline perfusion parameters and tumor response.

Leukocyte-associated Ig-like Receptor-1-deficient Mice Have an Altered Immune Cell Phenotype

Journal of Immunology (Baltimore, Md. : 1950). Jan, 2012  |  Pubmed ID: 22156345

Cross-linking of the collagen binding receptor leukocyte-associated Ig-like receptor-1 (LAIR-1) in vitro delivers an inhibitory signal that is able to downregulate activation-mediated signals. To study the in vivo function of LAIR-1, we generated LAIR-1(-/-) mice. They are healthy and fertile and have normal longevity; however, they show certain phenotypic characteristics distinct from wild-type mice, including increased numbers of splenic B, regulatory T, and dendritic cells. As LAIR-1(-/-) mice age, the splenic T cell population shows a higher frequency of activated and memory T cells. Because LAIR-1(+/+) and LAIR-1(-/-) T cells traffic with equal proficiency to peripheral lymphoid organs, this is not likely due to abnormal T lymphocyte trafficking. LAIR-1(-/-) mice have lower serum levels of IgG1 and, in response to T-dependent immunization with trinitrophenyl-OVA, switch less efficiently to Ag specific IgG2a and IgG2b, whereas switching to IgG1 is not affected. Several mouse disease models, including experimental autoimmune encephalitis and colitis, were used to examine the effect of LAIR-1 deficiency, and no differences in the responses of LAIR-1(-/-) and LAIR-1(+/+) mice were observed. Taken together, these observations indicate that LAIR-1 plays a role in regulating immune cells and suggest that any adverse effects of its absence may be balanced in vivo by other inhibitory receptors.

Maturational Time Course of the Envelope Following Response to Amplitude-modulated Acoustic Signals in Rats

International Journal of Audiology. Apr, 2012  |  Pubmed ID: 22176306

The maturation pattern of the envelope following response (EFR) was described using rats as an experimental model.

Investigating a High Ozone Episode in a Rural Mountain Site

Environmental Pollution (Barking, Essex : 1987). Mar, 2012  |  Pubmed ID: 22243863

A very high ozone episode with observed hourly values above 350 μg m(-3) occurred in July 2005 at the Lamas d'Olo air quality monitoring station, located in a mountainous area in the north of Portugal. Aiming to identify the origin and formation of this ozone-rich episode, a statistical analysis and a modelling approach were applied. A cross-spectrum analysis in the frequency domain and a synoptic analysis of the meteorological and air quality time series were performed. In order to go further in this analysis, a numerical modelling approach was applied. The results indicate that the transport of ozone and its precursors is the main responsible for the high ozone concentrations. Together with the local mountain breeze and subsidence conditions, the sea-breeze circulation transporting pollutants from the coastal urban and industrialized areas that reach the site during late afternoon turn out to be the driving forces for the ozone peaks.

Depressive Symptoms in Patients with Type 2 Diabetes Mellitus: Do Stress and Coping Matter?

Stress and Health : Journal of the International Society for the Investigation of Stress. Apr, 2012  |  Pubmed ID: 22282035

This article examines the relationship among diabetes-related stress, appraisal, coping and depressive symptoms in patients with type 2 diabetes mellitus (T2DM) using the transactional model of stress and coping (TMSC) as the theoretical framework. In this cross-sectional study, a convenience sample of 201 patients with T2DM was recruited from three outpatient clinics. Patients with depressive symptoms reported significantly more diabetes-related stress than patients without depressive symptoms. The results of path analysis suggest that patients who experience greater diabetes-related stress or greater depressive symptoms have a negative appraisal of their diabetes. Negative appraisal is, in turn, associated with greater use of avoidance, passive resignation and diabetes integration coping and lesser use of problem-focused coping. Avoidance, passive resignation and diabetes integration coping are, in turn, related to greater depressive symptoms or greater diabetes-related stress. Overall, the results of this study support the TMSC as a framework to elucidate the relationships among diabetes-related stress, appraisal, coping and depressive symptoms in patients with T2DM. However, given the cross-sectional nature of the study, we are unable to elucidate the directionality of the relationship between stress and depressive symptoms. Implications of the findings and the need for longitudinal studies to evaluate these relationships are discussed.

Baseline Susceptibility of Primary HIV-2 to Entry Inhibitors

Antiviral Therapy. Jan, 2012  |  Pubmed ID: 22293827

BACKGROUND: The baseline susceptibility of primary HIV-2 to maraviroc (MVC) and other entry inhibitors is currently unknown. METHODS: The susceptibility of 19 HIV-2 isolates obtained from asymptomatic and AIDS patients and seven HIV-1 clinical isolates to the fusion inhibitors enfuvirtide (ENF) and T-1249, and to the coreceptor antagonists AMD3100, TAK-779 and MVC, was measured using a TZM-bl cell-based assay. The 50% inhibitory concentration (IC(50)), 90% inhibitory concentration (IC(90)) and dose-response curve slopes were determined for each drug. RESULTS: ENF and T-1249 were significantly less active on HIV-2 than on HIV-1 (211- and 2-fold, respectively). AMD3100 and TAK-779 inhibited HIV-2 and HIV-1 CXCR4 tropic (X4) and CCR5 tropic (R5) variants with similar IC(50) and IC(90) values. MVC, however, inhibited the replication of R5 HIV-2 variants with significantly higher IC(90) values (42.7 versus 9.7 nM; P<0.0001) and lower slope values (0.7 versus 1.3; P<0.0001) than HIV-1. HIV-2 R5 variants derived from AIDS patients were significantly less sensitive to MVC than variants from asymptomatic patients, this being inversely correlated with the absolute number of CD4(+) T-cells. CONCLUSIONS: T-1249 is a potent inhibitor of HIV-2 replication indicating that new fusion inhibitors might be useful to treat HIV-2 infection. Coreceptor antagonists TAK-779 and AMD3100 are also potent inhibitors of HIV-2 replication. The reduced sensitivity of R5 variants to MVC, especially in severely immunodeficient patients, indicates that the treatment of HIV-2-infected patients with MVC might require higher dosages than those used in HIV-1 patients, and should be adjusted to the disease stage.

Human CD300a Binds to Phosphatidylethanolamine and Phosphatidylserine, and Modulates the Phagocytosis of Dead Cells

Blood. Mar, 2012  |  Pubmed ID: 22302738

CD300a is an immunoreceptor tyrosine-based inhibitory motif (ITIM) containing molecule that belongs to the CD300 family of paired activating/inhibitory receptors. It has been shown that its ligation inhibits activation signals on cells of both myeloid and lymphoid lineages. The ligands for CD300a have not been identified. Here, we show that a CD300a-Ig fusion protein specifically binds to apoptotic cells that are evolutionary apart, such as human and insect cells, suggesting that the ligand has to be conserved. Using surface plasmon resonance, ultracentrifugation, ELISA, and reporter cell assays, we identified phosphatidylethanolamine (PE) and phosphatidylserine (PS), 2 phospholipids that translocate to the outer leaflet of the plasma membrane of dead cells, as the ligands for CD300a. Mutational and structural modeling studies identified residues that are involved in the binding of CD300a to PE and PS and that form a cavity where the hydrophilic heads of PE and PS, can penetrate. CD300a down-regulates the uptake of apoptotic cells by macrophages and its ectopic expression in CD300a-negative cell lines also decreased the engulfment of dead cells. Collectively, our results indicate that PE and PS are ligands for CD300a, and that this interaction plays an important role in regulating the removal of dead cells.

Behavior of Natural Radionuclides in Surficial Sediments from an Estuary Impacted by Acid Mine Discharge and Industrial Effluents in Southwest Spain

Journal of Environmental Radioactivity. Feb, 2012  |  Pubmed ID: 22327046

The environmental degradation resulting from the acid mine drainage (AMD) and discharge from effluents of phosphogypsum (PG) piles in the watershed of Tinto and Odiel Rivers estuary over long periods of time has resulted in significant impact on the ecosystem of this estuary, resulting that the sediments are highly polluted by heavy metals and radionuclides from the discharge AMD and leachates from the PG. During resuspension of benthic sediments some of the radionuclides are desorbed making them bioavailable. In the present study, we investigate the spatial distribution of radionuclides U, Th and Ra and assess the factors and processes that caused the spatial distribution of these nuclides in this estuarine system. This study has global significance for other polluted environmental systems that are impacted by AMD and PG.

Association of X-linked Hydrocephalus and Hirschsprung Disease: Report of a New Patient with a Mutation in the L1CAM Gene

American Journal of Medical Genetics. Part A. Apr, 2012  |  Pubmed ID: 22344793

X-linked hydrocephalus (XLH) has an incidence of 1/30,000 male births and is characterized by intellectual disability, spastic paraplegia, adducted thumbs, and agenesis of corpus callosum, and/or corticospinal tract. The great proportion of cases is ascribed to loss of function mutations of L1CAM gene. Hirschsprung disease (HSCR) is characterized by the absence of ganglion cells in the submucosal and myenteric plexuses along a variable portion of the intestinal tract and has incidence of about 1/5,000. Although with several genes involved in its pathogenesis, the major HSCR gene is the RET proto-oncogene. To date only a few patients have been reported with both phenotypes and mutations in the L1CAM gene. In this report, we describe a new patient with concurrent XLH and HSCR. L1CAM mutational screening showed the presence of the G698R hemizygous mutation, which is a deleterious substitution affecting a key residue necessary for the correct folding of the protein. Moreover, the patient also carried the transcriptional enhancer RET mutation (c.73 + 9277T > C) in heterozygosis. We speculate that both the RET enhancer variant, and the L1CAM mutation may act in combination to produce the enteric phenotype, probably with the participation of other still unidentified molecular events. While it is obvious that additional studies are necessary to further delineate the association between XLH and HSCR in the presence of L1CAM mutations, the documentation of this new patient reinforces the role of this gene acting either in a direct or indirect way in the pathogenesis of Hirschsprung disease. © 2012 Wiley Periodicals, Inc.

Male and Female Differential Reproductive Rate Could Explain Parental Transmission Asymmetry of Mutation Origin in Hirschsprung Disease

European Journal of Human Genetics : EJHG. Mar, 2012  |  Pubmed ID: 22395866

Hirschsprung disease (HSCR, aganglionic megacolon) is a complex and heterogeneous disease with an incidence of 1 in 5000 live births. Despite the multifactorial determination of HSCR in the vast majority of cases, there is a monogenic subgroup for which private rare RET coding sequence mutations with high penetrance are found (45% of HSCR familial cases). An asymmetrical parental origin is observed for RET coding sequence mutations with a higher maternal inheritance. A parent-of-origin effect is usually assumed. Here we show that a differential reproductive rate for males and females also leads to an asymmetrical parental origin, which was never considered as a possible explanation till now. In the case of HSCR, we show a positive association between penetrance of the mutation and parental transmission asymmetry: no parental transmission asymmetry is observed in sporadic RET CDS mutation carrier cases for which penetrance of the mutation is low, whereas a parental transmission asymmetry is observed in affected sib-pairs for which penetrance of the mutation is higher. This allows us to conclude that the explanation for this parental asymmetry is that more severe mutations have resulted in a differential reproductive rate between male and female carriers.European Journal of Human Genetics advance online publication, 7 March 2012; doi:10.1038/ejhg.2012.35.

Tolerance Towards Dating Violence in Spanish Adolescents

Psicothema. May, 2012  |  Pubmed ID: 22420351

The study of intimate partner violence among adolescent and young couples in Spain remains unattended, although such abuses are well known and more frequent than in adulthood. The aim of this study is, on the one hand, to provide epidemiological information on dating relationships, and on the other hand, to identify attitudes towards violence. 2205 women enrolled in schools in diverse provinces of Spain, participated in the study. Average age was near 19 years (SD= 2.25). The Dating Violence Questionnaire (DVQ, in Spanish, CUVINO), a questionnaire that assesses both frequency and distress associated with violent behavior, was used. The DVQ allowed differentiating between groups of women self-labeled as abused and not abused on the basis of the frequency of sustained violence, although the levels of distress in the face of violence were statistically similar in both groups. Implications for future research and prevention programs are discussed.

MAX Mutations Cause Hereditary and Sporadic Pheochromocytoma and Paraganglioma

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Mar, 2012  |  Pubmed ID: 22452945

PURPOSE: Pheochromocytomas (PCC) and paragangliomas (PGL) are genetically heterogeneous neural crest-derived neoplasms. Recently we identified germline mutations in a new tumor suppressor susceptibility gene, MAX (MYC associated factor X), which predisposes carriers to PCC. How MAX mutations contribute to PCC/PGL and associated phenotypes remain unclear. This study aimed to examine the prevalence and associated phenotypic features of germline and somatic MAX mutations in PCC/PGL.Design: We sequenced MAX in 1694 patients with PCC or PGL (without mutations in other major susceptibility genes) from 17 independent referral centers. We screened for large deletions/duplications in 1535 patients using a multiplex polymerase chain reaction-based method. Somatic mutations were searched for in tumors from an additional 245 patients. The frequency and type of MAX mutation was assessed overall and by clinical characteristics.RESULTS: Sixteen MAX pathogenic mutations were identified in 23 index patients. All had adrenal tumors, including 13 bilateral or multiple PCCs within the same gland (P less than 0.001), 15.8% developed additional tumors at thoracic-abdominal sites, and 37% had familial antecedents. Age at diagnosis was lower (P=0.001) in MAX mutation carriers compared to non-mutated cases. Two patients (10.5%) developed metastatic disease. A mutation affecting MAX was found in five tumors, four of them confirmed as somatic (1.65%). MAX tumors were characterized by substantial increases in normetanephrine, associated with normal or minor increases in metanephrine.CONCLUSION: Germline mutations in MAX are responsible for 1.12% of PCC/PGL in patients without evidence of other known mutations, and should be considered in the genetic work-up of these patients.

Vulvar Intraepithelial Neoplasia: Evaluation of Treatment Modalities

Journal of Lower Genital Tract Disease. Mar, 2012  |  Pubmed ID: 22460275

OBJECTIVE: The study aimed to evaluate effectiveness and recurrence rate of vulvar intraepithelial neoplasia (VIN) treatment according to treatment modalities as follows: imiquimod (Aldara), laser ablation, laser excision, wide local excision, and skinning vulvectomy. It also aimed to analyze risk factors associated to VIN recurrence. MATERIALS AND METHODS: Between January 1997 and December 2010, 29 women were treated and followed up for VIN in our center. Demographics, risk factors, treatment modality, effectiveness, and recurrence data were recorded retrospectively. Study analysis used Student t test and χ test. RESULTS: The median age was 52 years (range = 22-77 years); 52% were smokers, 31% were immunosuppressed, and 34% had concomitant or previous lower genital tract dysplasia. Of all patients, 38% had laser ablation, 31% had laser excision, 24% had wide local excision, and 3% had vulvectomy and imiquimod, with 86.2% overall effectiveness and 20% recurrence (2 laser excision and 2 wide local excision), within a mean of 35 months. CONCLUSIONS: Excisional treatment has diagnostic and treatment advantages in VIN lesions. The goal is to prevent development of invasive vulvar cancer while preserving normal vulvar anatomy and function.

Impact of Forest Fires on Particulate Matter and Ozone Levels During the 2003, 2004 and 2005 Fire Seasons in Portugal

The Science of the Total Environment. Jan, 2012  |  Pubmed ID: 22088423

The main purpose of this work is to estimate the impact of forest fires on air pollution applying the LOTOS-EUROS air quality modeling system in Portugal for three consecutive years, 2003-2005. Forest fire emissions have been included in the modeling system through the development of a numerical module, which takes into account the most suitable parameters for Portuguese forest fire characteristics and the burnt area by large forest fires. To better evaluate the influence of forest fires on air quality the LOTOS-EUROS system has been applied with and without forest fire emissions. Hourly concentration results have been compared to measure data at several monitoring locations with better modeling quality parameters when forest fire emissions were considered. Moreover, hourly estimates, with and without fire emissions, can reach differences in the order of 20%, showing the importance and the influence of this type of emissions on air quality.

Early ROS-mediated DNA Damage and Oxidative Stress Biomarkers in Monoclonal B Lymphocytosis

Cancer Letters. Apr, 2012  |  Pubmed ID: 22115963

Monoclonal B Lymphocytosis (MBL) is defined as asymptomatic monoclonal B-cell expansion characterised by a CLL-phenotype, but with less than 5×10(9)/l circulating cells. Reactive oxygen species (ROS)-mediated cell damage plays a critical role in the initiation of carcinogenesis as well as in malignant transformation. The goal of this study was to perform an analysis of the oxidative stress statuses of patients affected by MBL and chronic lymphocytic leukaemia (CLL). We examined peripheral blood and urine specimens from 29 patients with MBL, 55 with CLL and 31 healthy subjects. There was a significant increase in the occurrence of the mutagenic base 8-oxo-2'-deoxiguanosine (8-oxo-dG) in the lymphocytes and urine of MBL and CLL patients compared with controls. Significant differences were also observed in the levels of the lipid peroxidation product malondialdehyde (MDA) and in the oxidised/reduced glutathione (GSSG/GSH) ratio, although an increase in 8-isoprostane was not detected. Interestingly, the antioxidant catalase activity of circulating lymphocytes decreased in the patient groups. In conclusion, early oxidative stress exists in patients with MBL and CLL, causing damage to DNA and lipid structures. The higher levels of 8-oxo-dG in lymphocytes than in urine may be related to a decrease in the capacity of DNA repair systems. There were no differences in the oxidative statuses of the MBL and CLL patients, suggesting that oxidative injuries appear during a pre-leukaemic state of the disease.