A comparative evaluation of endemic and non-endemic region of visceral leishmaniasis (Kala-azar) in India with ground survey and space technology

In visceral leishmaniasis, phlebotomine vectors are targets for control measures. Understanding the ecosystem of the vectors is a prerequisite for creating these control measures. This study endeavours to delineate the suitable locations of Phlebotomus argentipes with relation to environmental characteristics between endemic and non-endemic districts in India. A cross-sectional survey was conducted on 25 villages in each district. Environmental data were obtained through remote sensing images and vector density was measured using a CDC light trap. Simple linear regression analysis was used to measure the association between climatic parameters and vector density. Using factor analysis, the relationship between land cover classes and P. argentipes density among the villages in both districts was investigated. The results of the regression analysis indicated that indoor temperature and relative humidity are the best predictors for P. argentipes distribution. Factor analysis confirmed breeding preferences for P. argentipes by landscape element. Minimum Normalised Difference Vegetation Index, marshy land and orchard/settlement produced high loading in an endemic region, whereas water bodies and dense forest were preferred in non-endemic sites. Soil properties between the two districts were studied and indicated that soil pH and moisture content is higher in endemic sites compared to non-endemic sites. The present study should be utilised to make critical decisions for vector surveillance and controlling Kala-azar disease vectors.

Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.

Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes

A real-time polymerase chain reaction (PCR) assay with fluorescence resonance energy transfer (FRET) hybridisation probes combined with melting curve analysis was developed to detect Schistosoma japonicum in experimentally infected snails and in faecal samples of infected mice. This procedure is based on melting curve analysis of a hybrid between an amplicon from the S. japonicum internal transcribed spacer region 2 sequence, which is a 192-bp S. japonicum-specific sequence, and fluorophore-labelled specific probes. Real-time FRET PCR could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails and a single egg inoculated in 100 mg of non-infected mouse faeces. All S. japonicum-infected snails and all faecal samples from infected mice were positive. Non-infected snails, non-infected mouse faeces and genomic DNA from other parasites were negative. This assay is rapid and has potential for epidemiological S. japonicum surveys in snails, intermediate hosts and faecal samples of final hosts.

Detection of live and antibiotic-killed bacteria by quantitative real-time PCR of specific fragments of ribosomal RNA

RÉSUMÉ Le but d'un traitement antimicrobien est d'éradiquer une infection bactérienne. Cependant, il est souvent difficile d'en évaluer rapidement l'efficacité en utilisant les techniques standard. L'estimation de la viabilité bactérienne par marqueurs moléculaires permettrait d'accélérer le processus. Ce travail étudie donc la possibilité d'utiliser le RNA ribosomal (rRNA) à cet effet. Des cultures de Streptococcus gordonii sensibles (parent Wt) et tolérants (mutant Tol 1) à l'action bactéricide de la pénicilline ont été exposées à différents antibiotiques. La survie bactérienne au cours du temps a été déterminée en comparant deux méthodes. La méthode de référence par compte viable a été comparée à une méthode moléculaire consistant à amplifier par PCR quantitative en temps réel une partie du génome bactérien. La cible choisie devait refléter la viabilité cellulaire et par conséquent être synthétisée de manière constitutive lors de la vie de la bactérie et être détruite rapidement lors de la mort cellulaire. Le choix s'est porté sur un fragment du gène 16S-rRNA. Ce travail a permis de valider ce choix en corrélant ce marqueur moléculaire à la viabilité bactérienne au cours d'un traitement antibiotique bactéricide. De manière attendue, les S. gordonii sensibles à la pénicilline ont perdu ≥ 4 log10 CFU/ml après 48 heures de traitement par pénicilline alors que le mutant tolérant Tol1 en a perdu ≥ 1 log10 CFU/ml. De manière intéressant, la quantité de marqueur a augmenté proportionnellement au compte viable durant la phase de croissance bactérienne. Après administration du traitement antibiotique, l'évolution du marqueur dépendait de la capacité de la bactérie à survivre à l'action de l'antibiotique. Stable lors du traitement des souches tolérantes, la quantité de marqueur détectée diminuait de manière proportionnelle au compte viable lors du traitement des souches sensibles. Cette corrélation s'est confirmée lors de l'utilisation d'autres antibiotiques bactéricides. En conclusion, l'amplification par PCR du RNA ribosomal 16S permet d'évaluer rapidement la viabilité bactérienne au cours d'un traitement antibiotique en évitant le recours à la mise en culture dont les résultats ne sont obtenus qu'après plus de 24 heures. Cette méthode offre donc au clinicien une évaluation rapide de l'efficacité du traitement, particulièrement dans les situations, comme le choc septique, où l'initiation sans délai d'un traitement efficace est une des conditions essentielles du succès thérapeutique. ABSTRACT Assessing bacterial viability by molecular markers might help accelerate the measurement of antibiotic-induced killing. This study investigated whether ribosomal RNA (rRNA) could be suitable for this purpose. Cultures of penicillin-susceptible and penicillin-tolerant (Tol1 mutant) Streptococcus gordonii were exposed to mechanistically different penicillin and levofloxacin. Bacterial survival was assessed by viable counts, and compared to quantitative real-time PCR amplification of either the 16S-rRNA genes (rDNA) or the 16S rRNA, following reverse transcription. Penicillin-susceptible S. gordonii lost ≥ 4 log10 CFU/ml of viability over 48 h of penicillin treatment. In comparison, the Toll mutant lost ≤ 1 log10 CFU/ml. Amplification of a 427-base fragment of 16S rDNA yielded amplicons that increased proportionally to viable counts during bacterial growth, but did not decrease during drug-induced killing. In contrast, the same 427-base fragment amplified from 16S rDNA paralleled both bacterial growth and drug-induced killing. It also differentiated between penicillin-induced killing of the parent and the Toll mutant (≥4 log10 CFU/ml and ≤1 lo10 CFU/ml, respectively), and detected killing by mechanistically unrelated levofloxacin. Since large fragments of polynucleotides might be degraded faster than smaller fragments the experiments were repeated by amplifying a 119-base region internal to the origina1 427-base fragment. The amount of 119-base amplicons increased proportionally to viability during growth, but remained stable during drug treatment. Thus, 16S rRNA was a marker of antibiotic-induced killing, but the size of the amplified fragment was critical to differentiate between live and dead bacteria.

Disseny i construcció d'un motor 3D en temps real per la Gameboy Advance

El projecte desenvolupat ha tractat l’estudi i disseny d’un motor 3D interactiu a la consolaGame Boy Advance (GBA). La GBA disposa d’un processador ARM7TDMI a 16’78Mhz i no disposa de operacions 3D per-hardware, és una consola lenta en comparació lesque podem trobar al mercat d’avui en dia. Aquest treball, va partir de la construcció d’un prototipus ray-casting per-columna. Després,vàrem adaptar-lo a una estructura de portals i sectors. Més tard,es va introduir el mapeig de sostre/terra i de paisatges. Per últim,vàrem introduir efectes a la renderització per donar més realismeal recorregut del món, com il·luminació, objectes, etc.Tot i que es va estudiar l’arquitectura d’un motor eficient, no estenia prou per arribar a tenir un motor interactiu. Una de lestasques més difícils va ser la part de optimització. Peraconseguir-ho s’ha hagut de substituir operacions a temps realcostoses a temps de execució, replantejar parts de l’algorisme per fer-lo més eficient, entre altres

New tools for epidemiology: a space odyssey

Geographical Information Systems (GIS) facilitate access to epidemiological data through visualization and may be consulted for the development of mathematical models and analysis by spatial statistics. Variables such as land-cover, land-use, elevations, surface temperatures, rainfall etc. emanating from earth-observing satellites, complement GIS as this information allows the analysis of disease distribution based on environmental characteristics. The strength of this approach issues from the specific environmental requirements of those causative infectious agents, which depend on intermediate hosts for their transmission. The distribution of these diseases is restricted, both by the environmental requirements of their intermediate hosts/vectors and by the ambient temperature inside these hosts, which effectively govern the speed of maturation of the parasite. This paper discusses the current capabilities with regard to satellite data collection in terms of resolution (spatial, temporal and spectral) of the sensor instruments on board drawing attention to the utility of computer-based models of the Earth for epidemiological research. Virtual globes, available from Google and other commercial firms, are superior to conventional maps as they do not only show geographical and man-made features, but also allow instant import of data-sets of specific interest, e.g. environmental parameters, demographic information etc., from the Internet.

Visualització en temps real de multituds

Avui en dia, hi ha moltes pel.lícules o videojocs on apareixen grans quantitats de persones. Sovint és molt costós aconseguir grans quantitats de persones per realitzar les escenes, amb els problemes que això comporta (ja siguin econòmics o d’infraestructura) i la majoria de les vegades resulta inviable. Amb l’arribada de les noves targes gràfiques, és possible calcular aquesta visualització en temps real, donades una sèrie de simplificacions respecte de l’estructuració dels models a visualitzar.El que es pretén amb el GdM no és oferir una visualització molt realista de multituds (com ho faria el Massive Software, per exemple), sinó que el que es vol és la simulació de multituds usant molt menys temps per obtenir uns resultats prou bons per a simular el comportament i el moviment d' una multitud humana.Aquest projecte consta de dues parts: la primera, és la creació del model de persones que formaran la multitud. La segona, és la creació del GdM i la integració d’aquest model per la generació de la multitud

A continuous strain-space model of viral evolution within a host

Viruses rapidly evolve, and HIV in particular is known to be one of the fastest evolving human viruses. It is now commonly accepted that viral evolution is the cause of the intriguing dynamics exhibited during HIV infections and the ultimate success of the virus in its struggle with the immune system. To study viral evolution, we use a simple mathematical model of the within-host dynamics of HIV which incorporates random mutations. In this model, we assume a continuous distribution of viral strains in a one-dimensional phenotype space where random mutations are modelled by di ffusion. Numerical simulations show that random mutations combined with competition result in evolution towards higher Darwinian fitness: a stable traveling wave of evolution, moving towards higher levels of fi tness, is formed in the phenoty space.

Sin salud mental, no hay salud real

Boletín semanal para profesionales sanitarios de la Secretaría General de Salud Pública y Participación Social de la Consejería de Salud

Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR). A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

Hybrids and Regulation in the Global Political Economy

While regulation theory literature has made important contributions to the much-debated domain of globalisation by focusing on various aspects of post-Fordism, it has not yet fully engaged with the implications that can be drawn from critical approaches in international political economy. Recent studies have explored the transnational bases of new patterns and agents of change beyond states, firms and institutions traditionally involved in regulatory practices. Hybrid is often used as a default attribute reflecting lack of clear understanding of the breadth of this new type of influence and the opacity of the means involved. Drawing on the insights of philology and mythology, the paper argues that the notion of hybrid is relevant in elucidating the ontological ambiguity between imaginary and real aspects of globalisation. Furthermore, it specifies the categories involved in the analysis of emerging forms of hybrid regulation. Recent scholarship on globalisation tends to focus on the private-public nexus of the subjects involved in new forms of institutional arrangements and authority. Here, subjects, objects and space are analysed as joint issues. By focusing particularly on transformations affecting the role of the state, forms of competition, and their rescaling on a transnational basis, the concept of global hybrid is seen as complementary to the emancipation of regulation approaches from early emphasis on national levels of compromises.

Structural components in functional data

Analyzing functional data often leads to finding common factors, for which functional principal component analysis proves to be a useful tool to summarize and characterize the random variation in a function space. The representation in terms of eigenfunctions is optimal in the sense of L-2 approximation. However, the eigenfunctions are not always directed towards an interesting and interpretable direction in the context of functional data and thus could obscure the underlying structure. To overcome such difficulty, an alternative to functional principal component analysis is proposed that produces directed components which may be more informative and easier to interpret. These structural components are similar to principal components, but are adapted to situations in which the domain of the function may be decomposed into disjoint intervals such that there is effectively independence between intervals and positive correlation within intervals. The approach is demonstrated with synthetic examples as well as real data. Properties for special cases are also studied.

The interactional space in CLIL (Content and Language Integrated Learning) and EFL (English as a Foreign Language) classrooms

Aquest estudi pretén investigar els intercanvis verbals mestre/a – aprenent(s) en dos contextos d'instrucció diferents: classes amb un enfocament AICLE (Aprenentatge Integrat de Continguts Curriculars i Llengua Estrangera) on s’aprenen continguts no lingüístics a través de l’anglès, per una banda, i classes 'tradicionals' d'anglès com a llengua estrangera, on l’anglès és alhora objecte d’estudi i vehicle de comunicació, per una altra banda. Més concretament, les preguntes que formula el/la mestre/a, la producció oral dels aprenents i el 'feedback' del/de la mestre/a en els episodis d’atenció a la forma s’han estudiat a la llum de les principals teories provinents del camp de l’Adquisició de Segones Llengües (SLA) per tal de demostrar el seu paper en l’aprenentatge de l’anglès. El corpus de dades prové de l’enregistrament de 7 sessions AICLE i d'11 sessions EFL enregistrades en format àudio i vídeo en dos centres públics d’Educació Primària (EP) de Catalunya. A cadascuna de les escoles, el/la mateix/a mestre/a és l’encarregat/da dels dos tipus d’instrucció amb el mateix grup d’aprenents (10-11 anys d’edat), fet que permet eliminar variables individuals com l'aptitud dels aprenents o l'estil del/de la mestre/a.Els resultats mostren un cert nombre de similituds discursives entre AICLE i EFL donat que ambdós enfocaments tenen lloc en el context-classe amb unes característiques ben definides. Tal com apunta la recerca realitzada en aquest camp, la instrucció AICLE reuneix un seguit de condicions idònies per un major desenvolupament dels nivells de llengua anglesa més enllà de les classes ‘tradicionals’ d’anglès. Malgrat això, aquest estudi sembla indicar que el potencial d'AICLE pel que fa a facilitar una exposició rica a l’anglès i una producció oral significativa no s’explota degudament. En aquest sentit, els resultats d’aquest estudi poden contribuir a la formació dels futurs professors d'AICLE si es busca l’assoliment d’una complementarietat d’ambdós contextos amb l’objectiu últim de millorar els nivells de domini de la llengua anglesa.