Role of rs16944 (IL1B) and rs5743899 (TOLLIP) polymorphisms in the year-long presence of rhinovirus in the nostrils of healthy young volunteers

A infecção por rinovírus é uma das causas mais frequentes da constipação. Os mais afetados são crianças, idosos e portadores de doença respiratória crónica, que podem incorrer em doenças mais graves e complicações. Os rinovírus pertencem à família Picornaviridae sendo assim pequenos vírus de RNA de cadeia positiva com uma cápside proteica icosaédrica. Estes vírus são transmitidos muito facilmente atacando principalmente a mucosa nasal, mas a frequência e intensidade da infeção varia muito com os indivíduos e a época do ano. No presente trabalho procuramos caracterizar a influência de 2 polimorfismos em genes do sistema imunológico inato na infeção pelo rinovírus. Para tal, 89 voluntários (estudantes universitários) doaram mensalmente amostras nasais para a deteção rinovírus por RT-qPCR. Alguns destes indivíduos doaram também uma amostra de sangue (n=33) ou de epitélio bucal (n=15) para obter DNA genómico e caracterizar SNP’s dos genes TOLLIP e IL1B (rs5743899 e rs16944 respetivamente) por PCR-RFLP. Os resultados não revelaram qualquer associação do polimorfismo rs16944 (IL1B) e a presença de rinovírus nasal. Contudo todos os indivíduos que nunca testaram positivo para o rinovírus apresentaram o alelo G do polimorfismo rs5743899 (gene TOLLIP), sugerido um possível efeito deste alelo na eficaz eliminação imediata do rinovírus antes de ocorrer infeção viral. Contudo, o mesmo alelo apresentava também os valores mais elevados de títulos nasais de rinovírus, sugerindo que uma vez estabelecida a infeção estes indivíduos têm maior dificuldade em a controlar. Estes resultados estão em linha com estudos anteriores indicando que este alelo diminui a expressão de TOLLIP, potenciando a sinalização via TLR2, uma via particularmente importante no reconhecimento da cápside viral. Outros estudos revelaram ainda que o mesmo alelo está associado à diminuição da expressão de genes antivirais importantes como o IFN-λ1, o que pode potenciar uma menor eficácia de eliminação viral assim que a infeção viral ocorre.Rhinovirus infection is one of the most frequent causes of the common cold. Individuals most affected are children, elders and chronic respiratory patients who may present complications and a more severe, even life-threatening disease. Rhinoviruses belong to the Picornaviridae family thus being positive stranded RNA viruses with an icosahedral proteic capsid. These viruses are easily transmitted attacking mainly the nasal mucosa, but infection frequency varies considerably among individuals and with the season. In the present work we characterized the influence of two polymorphisms in innate immune system genes on rhinovirus infection. For that purpose, 89 voluntaries (university students) donated monthly nasal swabs for rhinovirus detection by RT-qPCR. Additionally, some individuals donated a blood sample (n=33) or buccal swab (n=15) for DNA extraction and SNP characterization of rs5743899 (TOLLIP gene) and rs16944 (IL1B gene) by PCR-RFLP. Results revealed no relation between rs16944 (IL1B) and rhinovirus nasal detection. However, every single individual that did never had rhinovirus detected in their nostril throughout the one year observation period showed the rs574388 G allele (TOLLIP gene), suggesting a possible effect of this allele in the efficacious prompt elimination of the virus before infection is attained. However, the same allele showed the highest viral titres, suggesting that once the infection is established, these individuals struggle to control it. These results are in agreement with previous studies indicating that the G allele decreases TOLLIP expression increasing TLR2 signalling, a particularly important route for RHV capsid detection. Other results also indicate that the same allele decreases antiviral gene expression (particularly IFN-λ1) which may potentiate a lower efficacy of viral clearance after infection is established

Role of rs16944 (IL1B) and rs5743899 (TOLLIP) polymorphisms in the year-long presence of rhinovirus in the nostrils of healthy young volunteers

A infecção por rinovírus é uma das causas mais frequentes da constipação. Os mais afetados são crianças, idosos e portadores de doença respiratória crónica, que podem incorrer em doenças mais graves e complicações. Os rinovírus pertencem à família Picornaviridae sendo assim pequenos vírus de RNA de cadeia positiva com uma cápside proteica icosaédrica. Estes vírus são transmitidos muito facilmente atacando principalmente a mucosa nasal, mas a frequência e intensidade da infeção varia muito com os indivíduos e a época do ano. No presente trabalho procuramos caracterizar a influência de 2 polimorfismos em genes do sistema imunológico inato na infeção pelo rinovírus. Para tal, 89 voluntários (estudantes universitários) doaram mensalmente amostras nasais para a deteção rinovírus por RT-qPCR. Alguns destes indivíduos doaram também uma amostra de sangue (n=33) ou de epitélio bucal (n=15) para obter DNA genómico e caracterizar SNP’s dos genes TOLLIP e IL1B (rs5743899 e rs16944 respetivamente) por PCR-RFLP. Os resultados não revelaram qualquer associação do polimorfismo rs16944 (IL1B) e a presença de rinovírus nasal. Contudo todos os indivíduos que nunca testaram positivo para o rinovírus apresentaram o alelo G do polimorfismo rs5743899 (gene TOLLIP), sugerido um possível efeito deste alelo na eficaz eliminação imediata do rinovírus antes de ocorrer infeção viral. Contudo, o mesmo alelo apresentava também os valores mais elevados de títulos nasais de rinovírus, sugerindo que uma vez estabelecida a infeção estes indivíduos têm maior dificuldade em a controlar. Estes resultados estão em linha com estudos anteriores indicando que este alelo diminui a expressão de TOLLIP, potenciando a sinalização via TLR2, uma via particularmente importante no reconhecimento da cápside viral. Outros estudos revelaram ainda que o mesmo alelo está associado à diminuição da expressão de genes antivirais importantes como o IFN-λ1, o que pode potenciar uma menor eficácia de eliminação viral assim que a infeção viral ocorre.Rhinovirus infection is one of the most frequent causes of the common cold. Individuals most affected are children, elders and chronic respiratory patients who may present complications and a more severe, even life-threatening disease. Rhinoviruses belong to the Picornaviridae family thus being positive stranded RNA viruses with an icosahedral proteic capsid. These viruses are easily transmitted attacking mainly the nasal mucosa, but infection frequency varies considerably among individuals and with the season. In the present work we characterized the influence of two polymorphisms in innate immune system genes on rhinovirus infection. For that purpose, 89 voluntaries (university students) donated monthly nasal swabs for rhinovirus detection by RT-qPCR. Additionally, some individuals donated a blood sample (n=33) or buccal swab (n=15) for DNA extraction and SNP characterization of rs5743899 (TOLLIP gene) and rs16944 (IL1B gene) by PCR-RFLP. Results revealed no relation between rs16944 (IL1B) and rhinovirus nasal detection. However, every single individual that did never had rhinovirus detected in their nostril throughout the one year observation period showed the rs574388 G allele (TOLLIP gene), suggesting a possible effect of this allele in the efficacious prompt elimination of the virus before infection is attained. However, the same allele showed the highest viral titres, suggesting that once the infection is established, these individuals struggle to control it. These results are in agreement with previous studies indicating that the G allele decreases TOLLIP expression increasing TLR2 signalling, a particularly important route for RHV capsid detection. Other results also indicate that the same allele decreases antiviral gene expression (particularly IFN-λ1) which may potentiate a lower efficacy of viral clearance after infection is established

Estimating traffic operations at multi-lane roundabouts: a case study

This paper addresses traffic modeling issues at urban multi-lane roundabouts where, despite circulating vehicles have priority, negotiation of the right-of-way can occur between antagonist traffic flows, as a result of minor drivers’ failing to obey the nominal operating rule (stop or yield control). Existing models for the estimation of operational performances have the shortcoming of not representing the interdependencies between entering and circulating vehicles at multi-lane roundabouts. An analytical capacity model derived from field observations was developed for this kind of intersections in a previous study. The complexity of the model lies in the difficulty of observing the behavioral parameters which are needed to implement the model. A procedure to get unknown behavioral parameters from traffic surveys is here proposed. This concerns saturation headways, often eluding direct observations due to rare occurrences of traffic conditions in which they can be observed. The unknown parameters were estimated through a regression model using on field data collected at a multi-lane roundabout. The presence of data correlation within a cluster of observations required the estimation of the regression parameters through a generalized estimating equation model. Results gave insight into the analysis of operations at multi-lane roundabouts, containing evidence to support assumptions made for the estimation of unobservable parameters

Patterning and dynamics of membrane adhesion under hydraulic stress

Hydraulic fracturing plays a major role in cavity formation during embryonic development, when pressurized fluid opens microlumens at cell-cell contacts, which evolve to form a single large lumen. However, the fundamental physical mechanisms behind these processes remain masked by the complexity and specificity of biological systems. Here, we show that adhered lipid vesicles subjected to osmotic stress form hydraulic microlumens similar to those in cells. Combining vesicle experiments with theoretical modelling and numerical simulations, we provide a physical framework for the hydraulic reconfiguration of cell-cell adhesions. We map the conditions for microlumen formation from a pristine adhesion, the emerging dynamical patterns and their subsequent maturation. We demonstrate control of the fracturing process depending on the applied pressure gradients and the type and density of membrane bonds. Our experiments further reveal an unexpected, passive transition of microlumens to closed buds that suggests a physical route to adhesion remodeling by endocytosis

Differences in chemical, physical and microbiological characteristics of Italian burrata cheeses made in artisanal and industrial plants of Apulia Region

The burrata cheese is a traditional product from Southern Italy, consisting of an envelope of pasta filata (stretched curd) filled with cream and pasta filata strips (usually leftovers from mozzarella production). Physical [water activity (aw), pH], chemical (moisture, NaCl content) and microbiological [total viable count (TVC), Listeria monocytogenes, Salmonella spp., Yersinia enterocolitica, Bacillus cereus, Escherichia coli, Enterobacteriaceae, coagulase-positive staphylococci] characteristics of burrata cheeses manufactured in artisanal and industrial plants were evaluated. The artisanal burrata showed lower aw values in the filling and the final product. The same was recorded in the filling for the moisture, probably due to differences between the types of cream used in the artisanal and the industrial cheesemaking. The pH value of the filling differed between the two groups but no difference was recorded in the final product. Microbiological differences were also recorded, with higher values for TVC and E. coli in artisanal than industrial burrata. All samples were negative for the other microbialdeterminations, with the exception of coagulase-positive staphylococci and Y. enterocolitica, which were detected in artisanal burrata. Differences in cheesemaking process were probably responsible for the strong variability of the physical and chemical data between the two cheeses; furthermore, differences in the hygienic features were also recorded. Even though artisanal products showed lower aw and pH values and higher NaCl concentration, the higher E. coli loads highlighted the need for a more accurate compliance with hygienic procedures along the artisanal cheesemaking process

Gene expression of stem cells at different stages of ontological human development

Objectives To compare multipotent mesenchymal stem cells (MSCs) obtained from chorionic villi (CV), amniotic fluid (AF) and placenta, with regard to their phenotype and gene expression, in order to understand if MSCs derived from different extra-embryonic tissues, at different stages of human ontological development, present distinct stemness characteristics. Study design MSCs obtained from 30 samples of CV, 30 of AF and 10 placentas (obtained from elective caesarean sections) were compared. MSCs at second confluence cultures were characterized by immunophenotypic analysis with flow cytometry using FACS CANTO II. The expression of the genes Oct-4 (Octamer-binding transcription factor 4, also known as POU5F1), Sox-2 (SRY box-containing factor 2), Nanog, Rex-1 (Zfp-42) and Pax-6 (Paired Box Protein-6), was analyzed. Real-time quantitative PCR was performed by ABI Prism 7700, after RNA isolation and retro-transcription in cDNA. Statistical analysis was performed using non-parametric test Kruskal–Wallis (XLSTAT 2011) and confirmed by REST software, to estimate fold changes between samples. Each gene was defined differentially expressed if p-value was <0.05. Results Cells from all samples were negative for haematopoietic antigens CD45, CD34, CD117 and CD33 and positive for the typical MSCs antigens CD13, CD73 and CD90. Nevertheless, MSCs from AF and placentas showed different fluorescence intensity, reflecting the heterogeneity of these tissues. The gene expression of OCT-4, SOX-2, NANOG was not significantly different among the three groups. In AF, REX-1 and PAX-6 showed a higher expression in comparison to CV. Conclusions MSCs of different extra-embryonic tissues showed no differences in immunophenotype when collected from second confluence cultures. The expression of OCT-4, NANOG and SOX-2 was not significantly different, demonstrating that all fetal sources are suitable for obtaining MSCs. These results open new possibilities for the clinical use of MSCs derived from easily accessible sources, in order to develop new protocols for clinical and experimental research

accounting for dispersion and correlation in estimating safety performance functions an overview starting from a case study

In statistical analysis of crash count data, as well as in estimating Safety Performance Functions (SPFs), the failure of Poisson equidispersion hypothesis and the temporal correlation in annual crash counts must be considered to improve the reliability of estimation of the parameters. After a short discussion on the statistical tools accounting for dispersion and correlation, the paper presents the methodological path followed in estimating a SPF for urban four-leg, signalized intersections. Since the case study exhibited signs of underdispersion, a Conway-Maxwell-Poisson Generalized Linear Model (GLM) was fitted to the data; then a quasi-Poisson model in the framework of Generalized Estimating Equations (GEEs) was performed in order to account for correlation. Results confirm that dispersion and correlation are phenomena that cannot be eluded in the estimation of SPFs under penalty of loss of efficiency in estimating model parameters. Generalized Estimating Equations overcome this problem allowing to incorporate together dispersion and temporal correlation when a quasi-Poisson distribution is used for modeling crash data. Moreover, whereas GEE regression is handy (many statistical software packages have already implemented GEE functions), the interest of COM-Poisson regression, because of difficulties in interpreting the model parameters and in arranging COM-Poisson codes, is still limited to the research field

Atmospheric and genetic parameters influence Rhinovirus transmission

info:eu-repo/semantics/publishedVersio

First evidence of intersex condition in extensively reared mullets from Sardinian lagoons (central–western Mediterranean, Italy)

The term intersex describes alterations in gonadal development with the simultaneous presence of male and female reproductive stages in the same gonad of a gonochoristic species. In coastal and estuarine environments, euryhaline fish living in polluted waters such as Mugilidae can frequently show these sexual anomalies. In this work, we analysed adult specimens of three species of euryhaline mullets (Chelon labrosus, Liza aurata and Mugil cephalus) from two Sardinian lagoons (Marceddì and San Teodoro) devoted to extensive aquacultural practices, in order to identify putative alterations in gonads and in gamete development. Overall, 13 of the 158 mullets examined (8.2%) were affected by gonadal disorders: four subjects (one C. labrosus, two L. aurata and one M. cephalus) exhibiting an intersex condition were found in the Marceddì lagoon and the other nine (five C. labrosus, two L. aurata and two M. cephalus) in the San Teodoro lagoon. Twelve of these gonads were classified as testis-ova (TOs) and one, belonging to a C. labrosus specimen, was a mixed gonadal tissue (MGT). Intersex condition was evaluated using an intersex index and all the recorded values showed a mild Ovotestis Severity Index (OSI). However, our findings suggest that fish gonadal disorders may be underestimated in extensive reared fish species, particularly in coastal brackish environments polluted by intensive agriculture and animal husbandry activities