Understanding consumer food choices & promotion of healthy and sustainable Mediterranean diet and lifestyle in children and adolescents through behavioural change actions: the DELICIOUS project.

Over the last decades, the abandonment of traditional dietary patterns, such as the Mediterranean diet, represents an important threat for human health and environmental safeguard. The DELICIOUS project aims to promote healthy lifestyles among children and adolescents by implementing activities and tools to increase the adherence to the Mediterranean Diet with an attention to the environmental impacts of the diet. This study protocol describes the DELICIOUS project as a single-arm, uncontrolled behavioural intervention providing formal and non-formal education activities, development of new snacks and recipe reformulation, web/mobile app development, and physical activities to school children and adolescents in five European countries. The project aims to increase awareness of the nutritional benefits and the sustainability aspects of the Mediterranean Diet and to promote consumers' empowerment through an online platform for sustainable and healthy meal planning in the school canteen.

Radiofrequency ablation combined with conductive fluid-based dopants (saline normal and colloidal gold): computer modeling and ex vivo experiments.

BACKGROUND: The volume of the coagulation zones created during radiofrequency ablation (RFA) is limited by the appearance of roll-off. Doping the tissue with conductive fluids, e.g., gold nanoparticles (AuNPs) could enlarge these zones by delaying roll-off. Our goal was to characterize the electrical conductivity of a substrate doped with AuNPs in a computer modeling study and ex vivo experiments to investigate their effect on coagulation zone volumes. METHODS: The electrical conductivity of substrates doped with normal saline or AuNPs was assessed experimentally on agar phantoms. The computer models, built and solved on COMSOL Multiphysics, consisted of a cylindrical domain mimicking liver tissue and a spherical domain mimicking a doped zone with 2, 3 and 4 cm diameters. Ex vivo experiments were conducted on bovine liver fragments under three different conditions: non-doped tissue (ND Group), 2 mL of 0.9% NaCl (NaCl Group), and 2 mL of AuNPs 0.1 wt% (AuNPs Group). RESULTS: The theoretical analysis showed that adding normal saline or colloidal gold in concentrations lower than 10% only modifies the electrical conductivity of the doped substrate with practically no change in the thermal characteristics. The computer results showed a relationship between doped zone size and electrode length regarding the created coagulation zone. There was good agreement between the ex vivo and computational results in terms of transverse diameter of the coagulation zone. CONCLUSIONS: Both the computer and ex vivo experiments showed that doping with AuNPs can enlarge the coagulation zone, especially the transverse diameter and hence enhance sphericity.

Speckle tracking echocardiography in the indeterminate form of Chagas disease.

BACKGROUND: Chagas disease is one of the most common diseases in Latin-America, and cardiac involvement is a significant cause of death. Assessment of myocardial strain may detect early myocardial damage. OBJECTIVES: To determine differences in longitudinal strain using speckle tracking to assess regional and global left ventricular function in patients with the indeterminate form of Chagas disease, in comparison with a control group. METHODS: This is a retrospective matched case-control study, conducted in a single center. We evaluated 45 adult patients with Chagas disease, diagnosed with 2 serological methods, without evidence of cardiac involvement, who were compared with 45 healthy control subjects, who were sex- and age-matched. All patients underwent Doppler echocardiography and longitudinal strain with speckle tracking. RESULTS: Median age was 59 years, and 60% were female. Echocardiographic parameters were similar in patients with Chagas and control subjects. In patients with Chagas, global strain differed significantly from that of control subjects (-17 vs -20.3, P < .001). Segmental strain showed 7 abnormal segments in patients with Chagas (P < .05). CONCLUSIONS: In patients with the indeterminate form of Chagas disease, global and segmental longitudinal peak systolic strain is reduced compared with healthy subjects, thus suggesting that it could be a sensitive technique to detect early myocardial damage. These findings could provide useful information regarding the pathophysiology of cardiac involvement and understand whether they might have prognostic usefulness or help develop strategies to modify the course and prognosis of patients with Chagas disease. A longitudinal prospective study would be necessary to validate our findings.

Mechanical dispersion in Fabry disease assessed with speckle tracking echocardiography.

BACKGROUND: Fabry disease (FD) is a rare X-linked storage disorder caused by deficiency of the lysosomal enzyme α-galactosidase A, and it typically causes multiorgan dysfunction. The main cause of death is heart disease resulting from left ventricular (LV) diastolic dysfunction, LV systolic dysfunction, severe LV hypertrophy (LVH), and sudden death. In several cardiac disorders, LV systolic dysfunction and ventricular arrhythmias are associated with mechanical dispersion (MD). MD has until now not been studied in patients with FD. OBJECTIVE: To investigate the prevalence of MD in patients with FD. METHODS: Complete echocardiographic data and speckle tracking echocardiographic data were collected. MD is an index of inter-segmental discoordination of contraction and is defined as the standard deviation (SD) of the time-to-peak longitudinal negative strain in 17 LV segments with a value >49 milliseconds. Patients with FD were divided into the following 2 groups: group I (patients with FD but no LVH, n = 64) and group II (patients with FD and LVH, n = 25). These groups were compared with a group of healthy subjects (group III, n = 50). Parametric variables were expressed as mean ± SD, and nonparametric variables were expressed as median and inter-quartile range. A P value <.05 was considered significant. RESULTS: A total of 113 patients with FD were included in this study. Of these, 24 (21%) were excluded because of poor imaging quality or presence of comorbidities, and the final study population consisted of 89 patients (mean age of 33.5 ± 14.5 years, 64% female). Group II patients were older than group I patients (46 ± 13 years vs 27 ± 11 years, P < .0001). There was no difference in LV ejection fraction between the 3 groups. There was also no difference in MD between groups I and III (32.4 ms [26-39] vs 32 ms [26-39]). In group II, the MD in 19 patients (76%) was 56 ms (39-80). CONCLUSIONS: To the best of our knowledge, this is the first study to assess the prevalence of MD in patients with FD. MD was observed in 76% of patients with FD and LVH. The use of MD in strain echocardiography may be beneficial in the assessment of patients with FD who develop heart failure.

Clinical significance of tissue Doppler imaging in chronic Chagas disease.

INTRODUCTION: Chagas disease (ChD) is one of the main parasitic diseases in Latin-America. Its heart involvement is the most important cause of death. The aim of this study is to evaluate if Doppler Tissue Imaging (DTI) may have a predictive value for later events in subjects with chronic ChD. METHODS: we analyses DTI variables of 543 patients with chronic ChD for the evaluation of predicting factors of events. Major adverse cardiovascular events (MACE) were considered as stroke, heart failure resistant to treatment, sustained ventricular tachycardia, implantable cardioverter-defibrillator, sudden death, and cardiovascular death. The following findings were also included in total evens: heart failure, bradycardia, ventricular arrhythmia, new conduction system abnormalities, and new echocardiographic abnormalities. Multivariate analysis with logistic regression was used in order to assess the Doppler and DTI parameters predicting events. Variables with a P-value ≤ .5 in the univariate analysis were included in the multivariate analysis. RESULTS: In patients with chronic ChD, the analysis of DTI parameters showed that S' wave and E' wave of the lateral wall of the left ventricle were significant predictors of MACE (OR: 0.83; 95% CI: 0.71-0.96; P-value: .015 and OR: 0.80; 95% CI: 0.66-0.98; P-value: .031, respectively). CONCLUSIONS: This study found that patients with chronic ChD who had events showed significantly lower parameters in the DTI. What is more, this study showed that even lower DTI parameters are significant predictors of events.

Correction to: Interaction of the heterotrimeric G protein alpha subunit SSG-1 of Sporothrix schenckii with proteins related to stress response and fungal pathogenicity using a yeast two-hybrid assay.

Following the publication of this article [1], it was brought to our attention that Fig. 7A lane 2 is identical to Fig. 7B lane 2 and Fig. 7B lane 4 is identical to Fig. 7C lane 4.

Identification of Immunoglobulin E-Binding Proteins of the Xerophilic Fungus Aspergillus penicillioides Crude Mycelial Mat Extract and Serological Reactivity Assessment in Subjects with Different Allergen Reactivity Profiles.

BACKGROUND: Aspergillus penicillioides is a very common indoor xerophilic fungus and potential causative agent of respiratory conditions. Although people are constantly exposed to A. penicillioides, no proteins with allergenic potential have been described. Therefore, we aim to confirm allergic sensitization to A. penicillioides through reactivity in serological assays and detect immunoglobulin E (IgE)-binding proteins. METHODS: In an indirect ELISA, we compared the serological reactivity to A. penicillioides between subjects with specific IgE (sIgE) (group 1, n = 54) and no sIgE reactivity (group 2, n = 15) against commercial allergens. Correlations and principal component analysis were performed to identify associations between reactivity to commercial allergens and A. penicillioides. IgE-binding proteins in A. penicillioides were visualized using Western blotting (WB) in group 1. The IgE-binding proteins with the highest reactivity were analyzed by mass spectrometry and confirmed by transcript matching. RESULTS: There was no statistical significance (p = 0.1656) between the study groups in serological reactivity. Correlations between reactivity to A. penicillioides, dog epithelia, Aspergillus fumigatus, and Penicillium chrysogenum were observed. WB experiments showed 6 IgE-binding proteins with molecular weights ranging from 45 to 145 kDa. Proteins of 108, 83, and 56 kDa showed higher reactivity. Mass spectrometry analysis of these 3 proteins led to the putative identification of NADP-specific glutamate dehydrogenase and catalase B. This was confirmed with transcriptome analysis. CONCLUSIONS: These results provide evidence of the presence of potential allergenic components in A. penicillioides. Further analysis of the putatively identified proteins should reveal their allergenic potential.

Passive cooling of cutaneous and subcutaneous tissues using phase changing materials: feasibility study using a numerical model.

In many dermatological applications, lowering the temperature of skin and maintaining specific temperatures for extended periods of time are fundamental requirements for treatment; for example, in targeting adipose tissue and managing cutaneous pain. In this work, we investigate the feasibility of using phase changing materials (PCMs) as an alternative passive, open-loop, heat extraction method for cooling cutaneous and subcutaneous tissues. We used a finite difference parametric approach to model the spatial and temporal progression of the heat transferred from the skin to a PCM in contact with the skin surface. We modelled the thermal performance of different PCMs, including different thicknesses. In addition, we used our model to propose application strategies. Numerical simulations demonstrate the feasibility of using PCMs for extracting heat from the skin and upper fat layers, inducing and maintaining similar temperatures as those induced by active closed-loop cooling with a cold plate. In terms of development, the critical design parameters are the temperature range of solidification of the material, the thickness of the material, and the rate of melting. Our study suggests that PCM-based devices may offer an alternative skin and adipose tissue cooling method that is simple to implement and use.

Asthma in British Columbia: Are we finally breathing easier? A population-based study of the burden of disease over 14 years.

OBJECTIVE: Asthma presents a significant global burden, but whether the incidence and prevalence of asthma is rising is still debated. The objective of this study was to determine the prevalence and incidence of asthma in British Columbia (BC), Canada, and characterize associated health services utilization. METHODS: We extracted data from provincial administrative hospitalization, medical services, and prescription drug databases for patients aged 5 to 55 years, during 1996 to 2009 having ≥270 MSP registration days and meeting asthma definition of: ≥1 hospital admissions with asthma as the principal diagnosis, or ≥2 physician visits for asthma as the principal diagnosis, or ≥3 asthma drug dispensings. Regression models were used to test change in asthma incidence and prevalence, and use of various health care services, such as physician and emergency department (ED) visits, and hospitalizations. RESULTS: 379,950 patients met the study criteria. The prevalence (2.6%) and incidence (0.7%) of asthma was relatively stable over the study period. There was a decline in proportion of patients visiting family practitioners (FP) (OR 0.92; 95% CI 0.90-0.94), specialists (OR 0.60; 95% CI 0.58-0.62), using ED services (OR 0.31; 95% CI 0.30-0.32) and hospitalizations (OR 0.34; 95% CI 0.31-0.37). Regional differences were noted, with lower rates of FP and specialist visits and higher rates of ED visits for asthma in rural versus urban areas. CONCLUSIONS: In BC, the incidence and prevalence of asthma has remained stable over 14 years. Although health service utilization declined, there is variation between rural and urban regions.

Oral morphine dosing predictions based on single dose in healthy children undergoing surgery.

BACKGROUND: Oral morphine has been proposed as an effective and safe alternative to codeine for after-discharge pain in children following surgery but there are few data guiding an optimum safe oral dose. AIMS: The aim of this study was to characterize the absorption pharmacokinetics of enteral morphine in order to simulate time-concentration profiles in children given common oral morphine dose regimens. METHODS: Children (2-6 years, n = 34) undergoing elective surgery and requiring opioid analgesia were randomized to receive preoperative oral morphine (100 mcg·kg-1 , 200 mcg·kg-1 , 300 mcg·kg-1 ). Blood sampling for morphine assay was performed at 30, 60, 90, 120, 180, and 240 min. Morphine serum concentrations were determined by liquid chromatography-mass spectroscopy and pharmacokinetic parameters were calculated using nonlinear mixed effects models. Current data were pooled with published time-concentration profiles from children (n = 1059, age 23 weeks postmenstrual age - 3 years) administered intravenous morphine, to determine oral bioavailability (F), absorption lag time (TLAG ), and absorption half-time (TABS ). These parameter estimates were used to predict concentrations in children given oral morphine (100, 200, 300, 400, 500 mcg·kg-1 ) at different dosing intervals (3, 4, 5, 6, 8, 12 h). RESULTS: The oral morphine formulation had F 0.298 (CV 36.5%), TLAG 0.45 (CV 63.6%) h and TABS 0.71 (CV 55%) h. A single-dose morphine 100 mcg·kg-1 achieved a mean CMAX 10 mcg·l-1 . Repeat 4-hourly dosing achieved mean steady-state concentration 13-18 mcg·l-1 ; concentrations associated with good analgesia after intravenous administration. Serum concentration variability was large ranging from 5 to 55 mcg·l-1 at steady state. CONCLUSIONS: Oral morphine 200 mcg·kg-1 then 100 mcg·kg-1 4 h or 150 mcg·kg-1 6 h achieves mean concentrations associated with analgesia. There was high serum concentration variability suggesting that respiration may be compromised in some children given these doses.

Performance Analysis of Cluster Formation in Wireless Sensor Networks.

Clustered-based wireless sensor networks have been extensively used in the literature in order to achieve considerable energy consumption reductions. However, two aspects of such systems have been largely overlooked. Namely, the transmission probability used during the cluster formation phase and the way in which cluster heads are selected. Both of these issues have an important impact on the performance of the system. For the former, it is common to consider that sensor nodes in a clustered-based Wireless Sensor Network (WSN) use a fixed transmission probability to send control data in order to build the clusters. However, due to the highly variable conditions experienced by these networks, a fixed transmission probability may lead to extra energy consumption. In view of this, three different transmission probability strategies are studied: optimal, fixed and adaptive. In this context, we also investigate cluster head selection schemes, specifically, we consider two intelligent schemes based on the fuzzy C-means and k-medoids algorithms and a random selection with no intelligence. We show that the use of intelligent schemes greatly improves the performance of the system, but their use entails higher complexity and selection delay. The main performance metrics considered in this work are energy consumption, successful transmission probability and cluster formation latency. As an additional feature of this work, we study the effect of errors in the wireless channel and the impact on the performance of the system under the different transmission probability schemes.

Study of the effect introduced by an integrating sphere on the temporal profile characterization of short laser pulses propagating through a turbid medium.

When a nanosecond laser pulse is transmitted through a highly scattering material, its irradiance decreases as it propagates; this is because of the spatial and temporal pulse profile stretching owing to multiple scattering events. Although the effect of temporal distortion is much less significant than that of the spatial distortion for applications where the laser beam is focused on a subsurface target (writing of waveguides, for example), it becomes significant for applications where the laser pulse must attain certain temporal width after the beam propagated is collimated through a turbid medium (photoacoustic tomography, for example). The objective of this work is to determine the transfer function associated to an integrating sphere measurement of the temporal intensity profile involving turbid media samples. The transfer function is found to be related to the geometrical characteristics of the integrating sphere and the optical properties of the turbid media. This procedure opens a new possibility for optical property characterization and enables the use of an integrating sphere for time-dependent intensity measurements.

Liver-specific case fatality due to chronic hepatitis C virus infection: a systematic review.

Despite reports that mortality is increasing, overall case fatality due to hepatitis C virus (HCV) is thought to be low. Given the variability in published rates, we aimed to synthesize estimates of liver-specific case fatality and all-cause mortality in chronic HCV according to follow-up duration, sustained viral response (SVR) to treatment, and liver disease severity. A systematic review was conducted of studies published in English from 2003 to 2013, reporting liver-specific case fatality estimates from HCV-infected samples. Thirty-five eligible articles were identified; 26 also presented estimates of all-cause mortality. Among community-based samples, liver-specific case fatality ranged from 0.3% over 5.7 years to 9.2% over 8.2 years of follow-up; and of all-cause mortality, from 4.0% over 5.7 years, to 23.0% over 8.2 years of follow-up. Estimates were higher among clinic-based samples and those with more severe liver disease. Among treated patients achieving SVR, liver-specific case fatality was low: up to 1.4% over 11.5 years of follow-up among samples with any severity of liver disease. Estimates were higher among those without SVR: up to 14.0% over 10 years of followup among samples with any severity of liver disease, and higher still among samples with more severe liver disease. The proportion of deaths attributable to liver-specific causes ranged from 55 to 85% among those with severe liver disease. Published estimates of fatality are high among certain populations of chronic HCV patients, with liver-specific causes being an important contributor. Understanding current HCV mortality rates is important for quantifying the total burden of HCV disease.

Characterization and ligand identification of a membrane progesterone receptor in fungi: existence of a novel PAQR in Sporothrix schenckii.

BACKGROUND: Adaptive responses in fungi result from the interaction of membrane receptors and extracellular ligands. Many different classes of receptors have been described in eukaryotic cells. Recently a new family of receptors classified as belonging to the progesterone-adiponectin receptor (PAQR) family has been identified. These receptors have the seven transmembrane domains characteristic of G-protein coupled receptors, but their activity has not been associated directly to G proteins. They share sequence similarity to the eubacterial hemolysin III proteins. RESULTS: A new receptor, SsPAQR1 (Sporothrix schenckii progesterone-adiponectinQ receptor1), was identified as interacting with Sporothrix schenckii G protein alpha subunit SSG-2 in a yeast two-hybrid assay. The receptor was identified as a member of the PAQR family. The cDNA sequence revealed a predicted ORF of 1542 bp encoding a 514 amino acids protein with a calculated molecular weight of 57.8 kDa. Protein domain analysis of SsPAQR1 showed the 7 transmembrane domains (TM) characteristic of G protein coupled receptors and the presence of the distinctive motifs that characterize PAQRs. A yeast-based assay specific for PAQRs identified progesterone as the agonist. S. schenckii yeast cells exposed to progesterone (0.50 mM) showed an increase in intracellular levels of 3', 5' cyclic adenosine monophosphate (cAMP) within the first min of incubation with the hormone. Different progesterone concentrations were tested for their effect on the growth of the fungus. Cultures incubated at 35°C did not grow at concentrations of progesterone of 0.05 mM or higher. Cultures incubated at 25°C grew at all concentrations tested (0.01 mM-0.50 mM) with growth decreasing gradually with the increase in progesterone concentration. CONCLUSION: This work describes a receptor associated with a G protein alpha subunit in S. schenckii belonging to the PAQR family. Progesterone was identified as the ligand. Exposure to progesterone increased the levels of cAMP in fungal yeast cells within the first min of incubation suggesting the connection of this receptor to the cAMP signalling pathway. Progesterone inhibited the growth of both the yeast and mycelium forms of the fungus, with the yeast form being the most affected by the hormone.

Suspected opioid overdose case resolved by CYP2D6 genotyping.

A 14-year-old female with suspected narcotic overdose had CYP2D6 genotyping performed to verify opiate intoxication. The role of pharmacogenetics in pain management and individualization of opiate pharmacotherapy is discussed.

Electrical-thermal performance of a cooled RF applicator for hepatic ablation with additional distant infusion of hypertonic saline: in vivo study and preliminary computer modeling.

PURPOSE: The Cool-tip electrode is one of the most widely employed applicators in radiofrequency (RF) hepatic ablation. Previous research demonstrated that it is possible to enlarge coagulation volume when the single cooled electrode is associated with distant infusion of saline (hybrid applicator). The aim of this study was to compare the electrical-thermal behaviour of the Cool-tip electrode with that of the hybrid applicator. MATERIALS AND METHODS: Forty-two RF ablations were performed on a total of 10 pigs: 22 with the Cool-tip electrode and 20 with the hybrid applicator (low infused saline volumetric flow rate of 6 mL/h at 2 mm distance). We compared both electrical performance (delivered power and number of roll-offs, i.e. sudden rises in impedance that interrupt the power delivery) and coagulation zone characteristics. In addition, we built a one-dimensional model to provide a basic physical explanation of the difference in performance between the different applicators. RESULTS: The experimental results showed that the number of roll-offs with the Cool-tip electrode was higher (24.3 ± 3.1 versus 6.7 ± 7.0). The hybrid applicator created larger coagulation volumes (19.7 ± 9.5 cm(3) versus 9.5 ± 5.8 cm(3)) with larger transverse diameters (2.5 ± 0.6 versus 1.9 ± 0.5 cm). The one-dimensional model confirmed the delay in the incidence of the first roll-off, but not the heterogeneity of the hybrid applicator's electrical performance in the experiments. CONCLUSIONS: The hybrid applicator produces fewer roll-off episodes than the Cool-tip electrode and creates larger coagulation volumes with larger transverse diameters.

Procedure to estimate thermophysical and geometrical parameters of embedded cancerous lesions using thermography.

Based on the fact that malignant cancerous lesions (neoplasms) develop high metabolism and use more blood supply than normal tissue, infrared thermography (IR) has become a reliable clinical technique used to indicate noninvasively the presence of cancerous diseases, e.g., skin and breast cancer. However, to diagnose cancerous diseases by IR, the technique requires procedures that explore the relationship between the neoplasm characteristics (size, blood perfusion rate and heat generated) and the resulting temperature distribution on the skin surface. In this research work the dual reciprocity boundary element method (DRBEM) has been coupled with the simulated annealing technique (SA) in a new inverse procedure, which coupled to the IR technique, is capable of estimating simultaneously geometrical and thermophysical parameters of the neoplasm. The method is of an evolutionary type, requiring random initial values for the unknown parameters and no calculations of sensitivities or search directions. In addition, the DRBEM does not require any re-meshing at each proposed solution to solve the bioheat model. The inverse procedure has been tested considering input data for simulated neoplasms of different sizes and positions in relation to the skin surface. The successful estimation of unknown neoplasm parameters validates the idea of using the SA technique and the DRBEM in the estimation of parameters. Other estimation techniques, based on genetic algorithms or sensitivity coefficients, have not been capable of obtaining a solution because the skin surface temperature difference is very small.

Bioinformatic Characterization and Molecular Evolution of the Lucina pectinata Hemoglobins.

(1) Introduction: Lucina pectinata is a clam found in sulfide-rich mud environments that has three hemoglobins believed to be responsible for the transport of hydrogen sulfide (HbILp) and oxygen (HbIILp and HbIIILp) to chemoautotrophic endosymbionts. The physiological roles and evolution of these globins in sulfide-rich environments are not well understood. (2) Methods: We performed bioinformatic and phylogenetic analyses with 32 homologous mollusk globin sequences. Phylogenetics suggests a first gene duplication resulting in sulfide binding and oxygen binding genes. A more recent gene duplication gave rise to the two oxygen-binding hemoglobins. Multidimensional scaling analysis of the sequence space shows evolutionary drift of HbIILp and HbIIILp, while HbILp was closer to the Calyptogena hemoglobins. Further corroboration is seen by conservation in the coding region of hemoglobins from L. pectinata compared to those from Calyptogena. (3) Conclusions: Presence of glutamine in position E7 in organisms living in sulfide-rich environments can be considered an adaptation to prevent loss of protein function. In HbILp a substitution of phenylalanine in position B10 is accountable for its unique reactivity towards H2S. It appears that HbILp has been changing over time, apparently not subject to functional constraints of binding oxygen, and acquired a unique function for a specialized environment.

Calcium/calmodulin kinase1 and its relation to thermotolerance and HSP90 in Sporothrix schenckii: an RNAi and yeast two-hybrid study.

BACKGROUND: Sporothrix schenckii is a pathogenic dimorphic fungus of worldwide distribution. It grows in the saprophytic form with hyaline, regularly septated hyphae and pyriform conidia at 25°C and as the yeast or parasitic form at 35°C. Previously, we characterized a calcium/calmodulin kinase in this fungus. Inhibitors of this kinase were observed to inhibit the yeast cell cycle in S. schenckii. RESULTS: The presence of RNA interference (RNAi) mechanism in this fungus was confirmed by the identification of a Dicer-1 homologue in S. schenckii DNA. RNAi technology was used to corroborate the role of calcium/calmodulin kinase I in S. schenckii dimorphism. Yeast cells were transformed with the pSilent-Dual2G (pSD2G) plasmid w/wo inserts of the coding region of the calcium/calmodulin kinase I (sscmk1) gene. Transformants were selected at 35°C using resistance to geneticin. Following transfer to liquid medium at 35°C, RNAi transformants developed as abnormal mycelium clumps and not as yeast cells as would be expected. The level of sscmk1 gene expression in RNAi transformants at 35°C was less than that of cells transformed with the empty pSD2G at this same temperature. Yeast two-hybrid analysis of proteins that interact with SSCMK1 identified a homologue of heat shock protein 90 (HSP90) as interacting with this kinase. Growth of the fungus similar to that of the RNAi transformants was observed in medium with geldanamycin (GdA, 10 µM), an inhibitor of HSP90. CONCLUSIONS: Using the RNAi technology we silenced the expression of sscmk1 gene in this fungus. RNAi transformants were unable to grow as yeast cells at 35°C showing decreased tolerance to this temperature. The interaction of SSCMK1 with HSP90, observed using the yeast two-hybrid assay suggests that this kinase is involved in thermotolerance through its interaction with HSP90. SSCMK1 interacted with the C terminal domain of HSP90 where effector proteins and co-chaperones interact. These results confirmed SSCMK1 as an important enzyme involved in the dimorphism of S. schenckii, necessary for the development of the yeast phase of this fungus. Also this study constitutes the first report of the transformation of S. schenckii and the use of RNAi to study gene function in this fungus.

Hepatitis C seroprevalence and correlation between viral load and viral genotype among primary care clients in Mexico.

OBJECTIVE: To measure hepatitis C virus (HCV) sero-prevalence, prevalence, hepatitis risk characteristics frequency, and genotype correlation with viral load among clients attending health care clinics. MATERIAL AND METHODS: Venous blood samples from l12 226 consecutive consenting adults were collected from January 2006 through December 2009. HCV antibodies were detected by immunoassay. HCV RNA was detected by qRT-PCR and viral genotype was performed by PCR and LIPA test. RESULTS: The HCV seroprevalence observed was l.5 % (C.I. 95% l.3-l.7), from seropositive individuals 60.9 % reported previous blood transfusion, 28.3% declared to have relatives with cirrhosis, 25.2% had tattoos or piercings, and 6.9% referred to have used drugs. Male gender and transfusion (p<0.001) were the most frequent hepatitis risk characteristics in the HCV seropositive group. Among seropositive subjects 48.3% presented HCV RNA.The most frequent genotype detected in all geographic areas of Mexico was l (subtype lA, 33%; subtype lB, 21.4%) followed by genotype 2 (subtype 2A, 8.50%). Subjects with genotype 1 had a significant correlation with the highest viral load. CONCLUSIONS: Our results show that nearly half of seropositive individuals are chronically infected. HCV infection has been shown in this study to be an emerging health problem in Mexico.