Influence of a light meal on routine haematological tests

Introduction Patient-related variables such as physical exercise stress and fasting status are Important sources of variability in laboratory testing However no clear indications about tasting requirements exist for routine haematological tests nor has the influence of meals been assessed Methods We studied 17 healthy volunteers who consumed a light meal containing a standardized amount of carbohydrates, protein and lipids Blood was taken for routine haematological tests before the meal and 1 2 and 4 hours thereafter Results One hour after the meal neutrophil count and mean corpuscular haemoglobin (MHC) increased significantly whereas lymphocyte and monocyte counts red blood cell distribution width, haematocrit, and mean corpuscular volume decreased significantly A clinically significant variation was only observed for lymphocytes Two hours after the meal a significant increase was observed for neutrophils and MCH whereas lymphocytes eosinophils, haemoglobin and haematocrit decreased significantly Clinically significant variations were recorded for lymphocytes red blood cells (RBC), haemoglobin haematocrit and MCH Four hours after the meal MCH was significantly increased while lymphocytes eosinophils, RBC, haemoglobin and haematocrit were significantly decreased Clinically significant variations were recorded for neutrophils eosinophils RBC hematocrit and MCH Conclusion The significant variation of several haematological parameters after a light meal demonstrates that the fasting time needs to be carefully considered in order to interpret the results of haematological tests correctly

Maternal immunization with ovalbumin prevents neonatal allergy development and up-regulates inhibitory receptor Fc gamma RIIB expression on B cells

Background: Preconception allergen immunization prevents neonatal allergen sensitization in mice by a complex interaction between regulatory cells/factors and antibodies. The present study assessed the influence of maternal immunization with ovalbumin (OVA) on the immune response of 3 day-old and 3 week-old offspring immunized or non-immunized with OVA and evaluated the effect of IgG treatment during fetal development or neonatal period. Results: Maternal immunization with OVA showed increased levels of Fc gamma RIIb expression in splenic B cells of neonates, which were maintained for up to 3 weeks and not affected by additional postnatal OVA immunization. Maternal immunization also exerted a down-modulatory effect on both IL-4 and IFN-gamma-secreting T cells and IL-4 and IL-12-secreting B cells. Furthermore, immunized neonates from immunized mothers showed a marked inhibition of antigen-specifc IgE Ab production and lowered Th2/Th1 cytokine levels, whereas displaying enhanced Fc gamma RIIb expression on B cells. These offspring also showed reduced antigen-specific proliferative response and lowered B cell responsiveness. Moreover, in vitro evaluation revealed an impairment of B cell activation upon engagement of B cell antigen receptor by IgG from OVA-immunized mice. Finally, in vivo IgG transference during pregnancy or breastfeeding revealed that maternal Ab transference was able to increase regulatory cytokines, such as IL-10, in the prenatal stage; yet only the postnatal treatment prevented neonatal sensitization. None of the IgG treatments induced immunological changes in the offspring, as it was observed for those from OVA-immunized mothers. Conclusion: Maternal immunization upregulates the inhibitory Fc gamma RIIb expression on offspring B cells, avoiding skewed Th2 response and development of allergy. These findings contribute to the advancement of prophylactic strategies to prevent allergic diseases in early life.

A Novel Hepatitis C Virus Genotyping Method Based on Liquid Microarray

The strategy used to treat HCV infection depends on the genotype involved. An accurate and reliable genotyping method is therefore of paramount importance. We describe here, for the first time, the use of a liquid microarray for HCV genotyping. This liquid microarray is based on the 5'UTR - the most highly conserved region of HCV - and the variable region NS5B sequence. The simultaneous genotyping of two regions can be used to confirm findings and should detect inter-genotypic recombination. Plasma samples from 78 patients infected with viruses with genotypes and subtypes determined in the Versant (TM) HCV Genotype Assay LiPA (version I; Siemens Medical Solutions, Diagnostics Division, Fernwald, Germany) were tested with our new liquid microarray method. This method successfully determined the genotypes of 74 of the 78 samples previously genotyped in the Versant (TM) HCV Genotype Assay LiPA (74/78, 95%). The concordance between the two methods was 100% for genotype determination (74/74). At the subtype level, all 3a and 2b samples gave identical results with both methods (17/17 and 7/7, respectively). Two 2c samples were correctly identified by microarray, but could only be determined to the genotype level with the Versant (TM) HCV assay. Genotype ""1'' subtypes (1a and 1b) were correctly identified by the Versant (TM) HCV assay and the microarray in 68% and 40% of cases, respectively. No genotype discordance was found for any sample. HCV was successfully genotyped with both methods, and this is of prime importance for treatment planning. Liquid microarray assays may therefore be added to the list of methods suitable for HCV genotyping. It provides comparable results and may readily be adapted for the detection of other viruses frequently co-infecting HCV patients. Liquid array technology is thus a reliable and promising platform for HCV genotyping.

The Effects of Commercially Available Preservative-Free FDA-Approved Triamcinolone (Triesence (R)) on Retinal Cells in Culture

Purpose: To evaluate the effects of Triesence (R) (TRI), a new preservative-free triamcinolone approved by the U. S. Food and Drug Administration (FDA) for intraocular use, on human retina pigment epithelial (ARPE-19) and rat neurosensory (R28) cells in culture. Methods: ARPE-19 and R28 cell cultures were treated 24 h with 1,000, 500, 200, or 100 mu g/mL of crystalline (cTRI) or 1,000, 500, or 200 mu g/mL of solubilized (sTRI). TRI was solubilized by centrifuging the drug, discarding the supernatant containing the vehicle and then resuspending the drug pellet in an equivalent amount of Dimethyl sulfoxide to achieve the same concentration as the commercial preparation. Percentage of cell viability (CV) was evaluated by a trypan blue dye-exclusion assay. The mitochondrial membrane potential (Delta Psi m) was analyzed with the JC-1 assay. The caspase-3/7 activity was measured by a fluorochrome assay. Results: In the ARPE-19 cultures, the cTRI caused a decrease in CV at 1,000 mg/mL (13.03 +/- 6.51; P < 0.001), 500 mu g/mL (28.87 +/- 9.3; P < 0.001), 200 mu g/mL (54.93 +/- 5.61; P < 0.001), and 100 mu g/mL (82.53 +/- 0.65; P < 0.005) compared with the untreated controls (96.98 +/- 0.16). In R28 cultures, the cTRI treatment also reduced CV values significantly (P < 0.001) for the 1,000 mu g/mL (22.73 +/- 2.44), 500 mu g/mL (34.63 +/- 1.91), 200 mu g/mL (58.70 +/- 1.39), and 100 mu g/m (75.33 +/- 2.47) compared with the untreated controls (86.08 +/- 3.54). Once the TRI was solubilized (sTRI), the CV and Delta Psi m remained similar to the untreated controls for both ARPE-19 and R28 cells. The sTRI treatment with 1,000, 500, and 200 mu g/mL increased in caspase-3/7 activity in ARPE-19 cells (P < 0.01) and in R28 cells (P < 0.05) compared with dimethyl sulfoxide equivalent controls. Conclusion: The crystalline form of TRI (cTRI) can cause a significant decrease in CV to cultured retinal cells. Once the TRI is solubilized (sTRI), at the same concentrations, the cells remain viable with no decrease in CV or Delta Psi m. The sTRI can, however, increase caspase-3/7 activity, thus suggesting some degree of apoptosis.

rst Transcriptional Activity Influences kirre mRNA Concentration in the Drosophila Pupal Retina during the Final Steps of Ommatidial Patterning

Background: Drosophila retinal architecture is laid down between 24-48 hours after puparium formation, when some of the still uncommitted interommatidial cells (IOCs) are recruited to become secondary and tertiary pigment cells while the remaining ones undergo apoptosis. This choice between survival and death requires the product of the roughest (rst) gene, an immunoglobulin superfamily transmembrane glycoprotein involved in a wide range of developmental processes. Both temporal misexpression of Rst and truncation of the protein intracytoplasmic domain, lead to severe defects in which IOCs either remain mostly undifferentiated and die late and erratically or, instead, differentiate into extra pigment cells. Intriguingly, mutants not expressing wild type protein often have normal or very mild rough eyes. Methodology/Principal Findings: By using quantitative real time PCR to examine rst transcriptional dynamics in the pupal retina, both in wild type and mutant alleles we showed that tightly regulated temporal changes in rst transcriptional rate underlie its proper function during the final steps of eye patterning. Furthermore we demonstrated that the unexpected wild type eye phenotype of mutants with low or no rst expression correlates with an upregulation in the mRNA levels of the rst paralogue kin-of-irre (kirre), which seems able to substitute for rst function in this process, similarly to their role in myoblast fusion. This compensatory upregulation of kirre mRNA levels could be directly induced in wild type pupa upon RNAi-mediated silencing of rst, indicating that expression of both genes is also coordinately regulated in physiological conditions. Conclusions/Significance: These findings suggest a general mechanism by which rst and kirre expression could be fine tuned to optimize their redundant roles during development and provide a clearer picture of how the specification of survival and apoptotic fates by differential cell adhesion during the final steps of retinal morphogenesis in insects are controlled at the transcriptional level.

Mast cell repopulation of the peritoneal cavity: contribution of mast cell progenitors versus bone marrow derived committed mast cell precursors

Background: Mast cells have recently gained new importance as immunoregulatory cells that are involved in numerous pathological processes. One result of these processes is an increase in mast cell numbers at peripheral sites. This study was undertaken to determine the mast cell response in the peritoneal cavity and bone marrow during repopulation of the peritoneal cavity in rats. Results: Two mast cell specific antibodies, mAb AA4 and mAb BGD6, were used to distinguish the committed mast cell precursor from more mature mast cells. The peritoneal cavity was depleted of mast cells using distilled water. Twelve hours after distilled water injection, very immature mast cells could be isolated from the blood and by 48 hours were present in the peritoneal cavity. At this same time the percentage of mast cells in mitosis increased fourfold. Mast cell depletion of the peritoneal cavity also reduced the total number of mast cells in the bone marrow, but increased the number of mast cell committed precursors. Conclusions: In response to mast cell depletion of the peritoneal cavity, a mast cell progenitor is released into the circulation and participates in repopulation of the peritoneal cavity, while the committed mast cell precursor is retained in the bone marrow.

Toxic and essential elements in blood from delivering women in selected areas of Sao Paulo State, Brazil

This study was designed to evaluate the degree of environmental contamination and possible exposure of pregnant women to toxic elements in seven selected areas of Sao Paulo State, Brazil. The overall median concentration of Mo in maternal blood was 0.53 mu g L(-1), highly significant differences found between sites (p < 0.0001). Cd was found to be low overall - 0.09 mu g L(-1) (0.01-0.58 mu g L(-1)) - with mothers from the Coastal and Rural 1 sites having the highest levels (p < 0.016). Median Hg concentration was 0.60 mu g L (1) (0.06 mu g L (1)-4.35 mu g L (1)); median Pb level was 16.2 mu g L (1) (3.5-57.7 mu g L(-1)) and no differences between sites were observed for both metals. Median Mn level was 16.7 mu g L(-1) (7.0-39.7 mu g L(-1)), being highest in Urban 2 site (p < 0.016). Concentrations of maternal Co were found to range between 0.06 mu g L(-1) and 1.1 mu g L(-1) (median 0.25 mu g L(-1)) and As level was 0.60 mu g L(-1) (0.10-3.8 mu g L(-1)) overall, with no statistical significance between sites for Co and As. Median Se concentrations were found to be 64 mg L(-1) (36-233 mu g L(-1)), with the highest median levels found in Urban 3 site; site differences were statistically significant (p < 0.0001). Correlation for each element (between paired maternal and cord blood) was measured only in Rural site 1; significant correlation was shown for Hg, Pb, Mn and Co (p < 0.05). These findings may be interpreted as indicating low environmental contamination in Sao Paulo State, Brazil. These findings could also indicate that pregnant women have little or no contact with pollutants, possibly due to awareness campaigns carried out by public health practitioners.

Antiretroviral resistance among HIV type 1-infected women first exposed to antiretrovirals during pregnancy: Plasma versus PBMCs

Resistance-associated mutations (RAMs) in plasma samples from HIV-1-infected women who received antiretroviral (ARV) prophylaxis during pregnancy was assessed and correlated with the detection of RAMs in peripheral blood mononuclear cells (PMBCs). The study population was composed of HIV-1-infected women enrolled in a prospective cohort study in Latin America and the Caribbean (NISDI Perinatal Study) as of March 1, 2005, who were diagnosed with HIV-1 infection during the current pregnancy, who received ARVs during pregnancy for prevention of mother-to-child transmission of HIV-1, and who were followed through at least the 6-12 week postpartum visit. Plasma samples collected at enrollment during pregnancy and at 6-12 weeks postpartum were assayed for RAMs. Plasma results were compared to previously described PBMC results from the same study population. Of 819 enrolled subjects, 197 met the eligibility criteria. Nucleic acid amplification was accomplished in 123 plasma samples at enrollment or 6-12 weeks postpartum, and RAMs were detected in 22 (17.9%; 95% CI: 11.7-25.9%). Previous analyses had demonstrated detection of RAMs in PBMCs in 19 (16.1%). There was high concordance between RAMs detected in plasma and PBMC samples, with only eight discordant pairs. The prevalence of RAMs among these pregnant, HIV-1-infected women is high (>15%). Rates of detection of RAMs in plasma and PBMC samples were similar.

A New Mouse Model for Marfan Syndrome Presents Phenotypic Variability Associated with the Genetic Background and Overall Levels of Fbn1 Expression

Marfan syndrome is an autosomal dominant disease of connective tissue caused by mutations in the fibrillin-1 encoding gene FBN1. Patients present cardiovascular, ocular and skeletal manifestations, and although being fully penetrant, MFS is characterized by a wide clinical variability both within and between families. Here we describe a new mouse model of MFS that recapitulates the clinical heterogeneity of the syndrome in humans. Heterozygotes for the mutant Fbn1 allele mg Delta(loxPneo), carrying the same internal deletion of exons 19-24 as the mg Delta mouse model, present defective microfibrillar deposition, emphysema, deterioration of aortic wall and kyphosis. However, the onset of a clinical phenotypes is earlier in the 129/Sv than in C57BL/6 background, indicating the existence of genetic modifiers of MFS between these two mouse strains. In addition, we characterized a wide clinical variability within the 129/Sv congenic heterozygotes, suggesting involvement of epigenetic factors in disease severity. Finally, we show a strong negative correlation between overall levels of Fbn1 expression and the severity of the phenotypes, corroborating the suggested protective role of normal fibrillin-1 in MFS pathogenesis, and supporting the development of therapies based on increasing Fbn1 expression.

In Vitro Adhesion of Streptococcus sanguinis to Dentine Root Surface After Treatment with Er:Yag Laser, Ultrasonic System, or Manual Curette

Objective: The purpose of this in vitro study was to evaluate the dentine root surface roughness and the adherence of Streptococcus sanguinis (ATCC 10556) after treatment with an ultrasonic system, Er:YAG laser, or manual curette. Background Data: Bacterial adhesion and formation of dental biofilm after scaling and root planing may be a challenge to the long-term stability of periodontal therapy. Materials and Methods: Forty flattened bovine roots were randomly assigned to one of the following groups: ultrasonic system (n = 10); Er:YAG laser (n = 10); manual curette (n = 10); or control untreated roots (n = 10). The mean surface roughness (Ra, mu m) of the specimens before and after exposure to each treatment was determined using a surface profilometer. In addition, S. sanguinis was grown on the treated and untreated specimens and the amounts of retained bacteria on the surfaces were measured by culture method. Results: All treatments increased the Ra; however, the roughest surface was produced by the curettes. In addition, the specimens treated with curettes showed the highest S. sanguinis adhesion. There was a significant positive correlation between roughness values and bacterial cells counts. Conclusion: S. sanguinis adhesion was the highest on the curette-treated dentine root surfaces, which also presented the greatest surface roughness.

Er : YAG laser, ultrasonic system, and curette produce different profiles on dentine root surfaces: An in vitro study

Objective: The aim of the present study was to compare the in vitro effects of the Er:YAG laser, an ultrasonic system, and manual curette on dentine root surface by roughness and micro-morphological analysis. Materials and Methods: Thirty-six flattened bovine roots were randomly assigned to one of the following groups: group 1 (n = 12): Er: YAG laser ( 2940 nm), 120 mJ/pulse, 10 Hz, 8.4 J/cm(2); group 2 ( n = 12): ultrasonic system; and group 3 ( n = 12): manual curette. The mean surface roughness (Ra) of each sample was measured using a profilometer before and after the treatments. The micro-morphology of the treated and untreated ( control) root surfaces was evaluated with scanning electron microscopy (SEM) at 50 x and 1000 x magnification. Results: Analysis with the profilometer showed that for equal times of instrumentation, the smoothest surfaces were produced by the Er: YAG laser and the ultrasonic system, followed by the curette ( p < 0.05). Morphological analyses demonstrated that treatment with the Er: YAG laser produced some areas with an irregular surface, craters, and ablation of the intertubular dentin. The smear layer was removed and dentine tubules were opened by both curettes and the ultrasonic system. The micro-morphology of the dentine root surface after ultrasonic treatment, however, demonstrated randomly distributed areas cratering. Conclusion: All instruments increased the roughness of the dentine root surface after treatment; however, the curette produced rougher surfaces than the other devices. SEM analysis revealed distinct root surface profiles produced by the three devices.

FAM5C Contributes to Aggressive Periodontitis

Aggressive periodontitis is characterized by a rapid and severe periodontal destruction in young systemically healthy subjects. A greater prevalence is reported in Africans and African descendent groups than in Caucasians and Hispanics. We first fine mapped the interval 1q24.2 to 1q31.3 suggested as containing an aggressive periodontitis locus. Three hundred and eighty-nine subjects from 55 pedigrees were studied. Saliva samples were collected from all subjects, and DNA was extracted. Twenty-one single nucleotide polymorphisms were selected and analyzed by standard polymerase chain reaction using TaqMan chemistry. Non-parametric linkage and transmission distortion analyses were performed. Although linkage results were negative, statistically significant association between two markers, rs1935881 and rs1342913, in the FAM5C gene and aggressive periodontitis (p = 0.03) was found. Haplotype analysis showed an association between aggressive periodontitis and the haplotype A-G (rs1935881-rs1342913; p = 0.009). Sequence analysis of FAM5C coding regions did not disclose any mutations, but two variants in conserved intronic regions of FAM5C, rs57694932 and rs10494634, were found. However, these two variants are not associated with aggressive periodontitis. Secondly, we investigated the pattern of FAM5C expression in aggressive periodontitis lesions and its possible correlations with inflammatory/immunological factors and pathogens commonly associated with periodontal diseases. FAM5C mRNA expression was significantly higher in diseased versus healthy sites, and was found to be correlated to the IL-1 beta, IL-17A, IL-4 and RANKL mRNA levels. No correlations were found between FAM5C levels and the presence and load of red complex periodontopathogens or Aggregatibacter actinomycetemcomitans. This study provides evidence that FAM5C contributes to aggressive periodontitis.

On the mass determination of super-Earths orbiting active stars: the CoRoT-7 system

Context. Star activity makes the mass determination of CoRoT-7b and CoRoT 7c uncertain. Investigators of the CoRoT team proposed several solutions, but all but one of them are larger than the initial determinations of 4.8 +/- 0.8 M(Earth) for CoRoT-7b and 8.4 +/- 0.9 M(Earth) for CoRoT 7c. Aims. This investigation uses the excellent HARPS radial velocity measurements of CoRoT-7 to redetermine the planet masses and to explore techniques for determining mass and orbital elements of planets discovered around active stars when the relative variation in the radial velocity due to the star activity cannot be considered as just noise and can exceed the variation due to the planets. Methods. The main technique used here is a self-consistent version of the high-pass filter used by Queloz et al. (2009, A&A, 506, 303) in the first mass determination of CoRoT-7b and CoRoT-7c. The results are compared to those given by two alternative techniques: (1) the approach proposed by Hatzes et al. (2010, A&A, 520, A93) using only those nights in which two or three observations were done; (2) a pure Fourier analysis. In all cases, the eccentricities are taken equal to zero as indicated by the study of the tidal evolution of the system. The periods are also kept fixed at the values given by Queloz et al. Only the observations done in the time interval BJD 2 454 847-873 are used because they include many nights with multiple observations; otherwise, it is not possible to separate the effects of the rotation fourth harmonic (5.91 d = P(rot)/4) from the alias of the orbital period of CoRoT-7b (0.853585 d). Results. The results of the various approaches are combined to give planet mass values 8.0 +/- 1.2 M(Earth) for CoRoT-7b and 13.6 +/- 1.4 M(Earth) for CoRoT 7c. An estimation of the variation of the radial velocity of the star due to its activity is also given. Conclusions. The results obtained with three different approaches agree to give higher masses than those in previous determinations. From the existing internal structure models they indicate that CoRoT-7b is a much denser super-Earth. The bulk density is 11 +/- 3.5 g cm(-3), so CoRoT-7b may be rocky with a large iron core.

Searching for star formation outside galaxies: Multiwavelength analysis of the intragroup medium of Hickson Compact Group 100

We use multiwavelength data (H I, FUV, NUV, R) to search for evidence of star formation in the intragroup medium of the Hickson Compact Group 100. We find that young star-forming regions are located in the intergalactic H I clouds of the compact group which extend to over 130 kpc away from the main galaxies. A tidal dwarf galaxy (TDG) candidate is located in the densest region of the H I tail, 61 kpc from the brightest group member and its age is estimated to be only 3.3 Myr. Fifteen other intragroup H II regions and TDG candidates are detected in the Galaxy Evolution Explorer (GALEX) FUV image and within a field 10' x 10' encompassing the H I tail. They have ages <200 Myr, H I masses of 10(9.2-10.4) M(circle dot), 0.001 densities about one order of magnitude lower than N(H I) similar to 10(21) cm(-2).

Transiting exoplanets from the CoRoT space mission XVI. CoRoT-14b: an unusually dense very hot Jupiter

In this paper, the CoRoT Exoplanet Science Team announces its 14th discovery. Herein, we discuss the observations and analyses that allowed us to derive the parameters of this system: a hot Jupiter with a mass of 7.6 +/- 0.6 Jupiter masses orbiting a solar-type star (F9V) with a period of only 1.5 d, less than 5 stellar radii from its parent star. It is unusual for such a massive planet to have such a small orbit: only one other known higher mass exoplanet orbits with a shorter period.