Low Molecular Weight Chitosan (LMWC)-based Polyplexes for pDNA Delivery: From Bench to Bedside

Non-viral gene delivery vectors are emerging as a safer alternative to viral vectors. Among natural polymers, chitosan (Ch) is the most studied one, and low molecular weight Ch, specifically, presents a wide range of advantages for non-viral pDNA delivery. It is crucial to determine the best process for the formation of Low Molecular Weight Chitosan (LMWC)-pDNA complexes and to characterize their physicochemical properties to better understand their behavior once the polyplexes are administered. The transfection efficiency of Ch based polyplexes is relatively low. Therefore, it is essential to understand all the transfection process, including the cellular uptake, endosomal escape and nuclear import, together with the parameters involved in the process to improve the design and development of the non-viral vectors. The aim of this review is to describe the formation and characterization of LMWC based polyplexes, the in vitro transfection process and finally, the in vivo applications of LMWC based polyplexes for gene therapy purposes.

Exploring new labelling strategies for boronated compounds: towards fast development and efficient assessment of BNCT drug candidates

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Biologie de l'Ethmalose, Ethmalosa fimbriata (Bowdich), en C��te d'Ivoire. 1 - Description de la reproduction et des premiers stades larvaires

Descriptions of spawning and larval development of Ethmalosa, up to the vitelline vesicle resorption stage, are made from plankton samplings in the Ebri�� coastal lagoon and from artificially fertilized eggs. Spawning takes place from November to June in waters with salinities of 18 to 26 parts per thousand, and temperatures of 22.8 to 30.2 degrees, for 13-14 cm long fishes.

Ecologia molecular de ostras (Crassostrea spp.) do Atl��ntico Tropical

Crassostrea (Sacco, 1897) �� o g��nero mais importante do mundo de ostras de cultivo e consiste de 34 esp��cies distribu��das pelas regi��es tropicais e temperadas do globo. C. gasar e C. rhizophorae s��o as duas esp��cies nativas que est��o distribu��das ao longo de toda a costa do Brasil at�� o Caribe. C. gasar tamb��m ocorre na costa da Africa. Ainda que sua distribui����o seja extensa e com disponibilidade abundante, o cultivo de ostras nativas no Brasil ainda �� incipiente e a delimita����o correta dos estoques mant��m-se incerta. O sucesso do desenvolvimento da malacocultura, que �� recomendada internacionalmente como forma sustent��vel de aquicultura, depende da resolu����o desses problemas. Assim, com o objetivo de determinar geneticamente seus estoques no Atl��ntico como tamb��m estimar sua hist��ria demogr��fica, dois diferentes marcadores moleculares foram empregados: sequ��ncias de DNA da regi��o controle mitocondrial e loci de microssat��lites esp��cie-especif��cos, desenvolvidos no presente estudo. Foram sequenciados fragmentos da regi��o controle de um total de 930 indiv��duos de C. gasar e C. rhizophorae coletados em 32 localidades que inclu��ram o Caribe, a Guiana Francesa, a costa brasileira e a ��frica. Tamb��m foram realizadas genotipagens de 1178 indiv��duos, e ambas as esp��cies, com 9 e 11 loci de microssat��lites para C. gasar e C. rhizophorae, respectivamente. Os dados gen��ticos foram analisados atrav��s de diferentes abordagens (��ndices de estrutura����o (FST) e de (Jost D), an��lise molecular de vari��ncia (AMOVA), an��lise espacial molecular de vari��ncia (SAMOVA), Bayesian Skyline Plots (BSP), an��lise fatorial de correspond��ncia (AFC) e an��lise de atribui����o Bayesiana (STRUCTURE)). Os resultados indicaram um padr��o geral de estrutura����o, onde dois diferentes estoques foram detectados para ambas as esp��cies: grupos do norte e do sul, onde o Rio de Janeiro seria a regi��o limitante entre os dois estoques. Os maiores valores dos ��ndices de estrutura����o foram encontrados para C. gasar, indicando que esta esp��cie estaria mais estruturada do que C. rhizophorae. As an��lises demogr��ficas indicaram uma prov��vel expans��o das popula����es durante o ultimo per��odo glacial e uma poss��vel origem americana das popula����es africanas. Todos os resultados sugeriram a exist��ncia de uma barreira geogr��fica pr��xima ao Rio de Janeiro, que poderia ser a cadeia de Vit��ria-Trindade e o fen��meno de ressurg��ncia que ocorre em Cabo Frio (RJ). Esses resultados ser��o de grande utilidade para estabelecer crit��rios para sele����o de sementes para cultivo ao longo da costa do Brasil que permitir�� o manejo adequado dos estoques ostre��colas, prevenindo seu desaparecimento como j�� ocorrido em outros recifes no mundo.

Stock evaluation and development of a breeding program for common carp (Cyprinus carpio) in Karnataka, India: progress of a research project

Common carp (Cyprinus carpio) is the single most important species for aquaculture in the state of Karnataka, India, where it is generally grown in polyculture with Indian major carps. Precocious maturation and unwanted reproduction in the species have been identified as constraints to increase production in aquaculture and culture-based fisheries in Karnataka state. Stocks of C. carpio obtained from Hungary (Amur and P3), Indonesia (Rajdanu) and Vietnam (SV) are being assessed alongside two local stocks (L-BRP and L-FRS) in a series of culture performance trials with the objective of setting up a base population for selective breeding. The paper presents progress of research being undertaken at the Fisheries Research Station, University of Agricultural Sciences, Bangalore, India.

Early response to nanoparticles in the Arabidopsis transcriptome compromises plant defence and root-hair development through salicylic acid signalling

Background: The impact of nano-scaled materials on photosynthetic organisms needs to be evaluated. Plants represent the largest interface between the environment and biosphere, so understanding how nanoparticles affect them is especially relevant for environmental assessments. Nanotoxicology studies in plants allude to quantum size effects and other properties specific of the nano-stage to explain increased toxicity respect to bulk compounds. However, gene expression profiles after exposure to nanoparticles and other sources of environmental stress have not been compared and the impact on plant defence has not been analysed. Results: Arabidopsis plants were exposed to TiO2-nanoparticles, Ag-nanoparticles, and multi-walled carbon nanotubes as well as different sources of biotic (microbial pathogens) or abiotic (saline, drought, or wounding) stresses. Changes in gene expression profiles and plant phenotypic responses were evaluated. Transcriptome analysis shows similarity of expression patterns for all plants exposed to nanoparticles and a low impact on gene expression compared to other stress inducers. Nanoparticle exposure repressed transcriptional responses to microbial pathogens, resulting in increased bacterial colonization during an experimental infection. Inhibition of root hair development and transcriptional patterns characteristic of phosphate starvation response were also observed. The exogenous addition of salicylic acid prevented some nano-specific transcriptional and phenotypic effects, including the reduction in root hair formation and the colonization of distal leaves by bacteria. Conclusions: This study integrates the effect of nanoparticles on gene expression with plant responses to major sources of environmental stress and paves the way to remediate the impact of these potentially damaging compounds through hormonal priming.

HPV infection, a described cause for human cancer: HPV detection and molecular mechanisms of HPV carcinogenesis (Review).

[en]Human papillomavirus (HPV) belongs to the Papillomaviridae virus family and it is one of the most common sexual transmission infections. HPV genome is composed of eight genes, including two early genes and six late genes. Among these late genes, E6 and E7 code for proteins that trigger cell-cycle re-entry in infected cells, which can lead to cervical cancer development. The IARC (International Agency for Research Cancer) proposed a guideline based on Hill���s criteria to determine whether the relation between HPV infection and cervical cancer is causal or not. Epidemiological studies have demonstrated that HPV infection is a necessary but non-sufficient cause for cervical cancer. Furthermore, HPV infection is considered the first necessary cause described of a human cancer, being HPV16 and 18 carcinogenic to humans and the most studied types. Cervical cancer is the second leading cause of cancer death among women worldwide. Different screening programs are carried out with the aim of preventing cervical cancer; such as cytologies and HPV tests. There are two main methods which are equally usable to detect HPV: the real-time PCR assays and the array assays. Regarding the molecular mechanisms of HPV mediated malignancies, E2, E6 and E7 proteins of HPV16 lead to immune response evasion, inducing IL-10 and TGF-��1 gene expression. Besides, E6 and E7 proteins allow cell-cycle reentry, phosphorylating RB and ubiquitinating p53 respectively. HPV genome integration in host genome leads to the alteration of host and viral genes expression, including oncogenes and tumor suppressor genes. However, the differences of E6 and E7 oncoproteins in different HPV types is poorly known due to the fact that almost the most studied HPV type has been HPV16.

LDLR and PCSK9 Are Associated with the Presence of Antiphospholipid Antibodies and the Development of Thrombosis in aPLA Carriers

Introduction The identification of the genetic risk factors that could discriminate non-thrombotic from thrombotic antiphospholipid antibodies (aPLA) carriers will improve prognosis of these patients. Several human studies have shown the presence of aPLAs associated with atherosclerotic plaque, which is a known risk factor for thrombosis. Hence, in order to determine the implication of atherosclerosis in the risk of developing thrombosis in aPLA positive patients, we performed a genetic association study with 3 candidate genes, APOH, LDLR and PCSK9. Material & Methods For genetic association study we analyzed 190 aPLA carriers -100 with non-thrombotic events and 90 with thrombotic events-and 557 healthy controls. Analyses were performed by chi(2) test and were corrected by false discovery rate. To evaluate the functional implication of the newly established susceptibility loci, we performed expression analyses in 86 aPLA carrier individuals (43 with thrombotic manifestations and 43 without it) and in 45 healthy controls. Results Our results revealed significant associations after correction in SNPs located in LDLR gene with aPLA carriers and thrombotic aPLA carriers, when compared with healthy controls. The most significant association in LDLR gene was found between SNP rs129083082 and aPLA carriers in recessive model (adjusted P-value = 2.55 x 10(-3); OR = 2.18; 95% CI = 1.49-3.21). Furthermore, our work detected significant allelic association after correction between thrombotic aPLA carriers and healthy controls in SNP rs562556 located in PCSK9 gene (adjusted P-value = 1.03 x 10(-2); OR = 1.60; 95% CI = 1.24-2.06). Expression level study showed significantly decreased expression level of LDLR gene in aPLA carriers (P-value < 0.0001; 95% CI 0.16-2.10; SE 0.38-1.27) in comparison to the control group. Discussion Our work has identified LDLR gene as a new susceptibility gene associated with the development of thrombosis in aPLA carriers, describing for the first time the deregulation of LDLR expression in individuals with aPLAs. Besides, thrombotic aPLA carriers also showed significant association with PCSK9 gene, a regulator of LDLR plasma levels. These results highlight the importance of atherosclerotic processes in the development of thrombosis in patients with aPLA.

Identification of Sex and Female���s Reproductive Stage in Commercial Fish Species through the Quantification of Ribosomal Transcripts in Gonads

The estimation of maturity and sex of fish stocks in European waters is a requirement of the EU Data Collection Framework as part of the policy to improve fisheries management. On the other hand, research on fish biology is increasingly focused in molecular approaches, researchers needing correct identification of fish sex and reproductive stage without necessarily having in house the histological know-how necessary for the task. Taking advantage of the differential gene transcription occurring during fish sex differentiation and gametogenesis, the utility of 5S ribosomal RNA (5S rRNA) and General transcription factor IIIA (gtf3a) in the molecular identification of sex and gametogenic stage was tested in different economically-relevant fish species from the Bay of Biscay. Gonads of 9 fish species (, Atlantic, Atlantic-chub and horse mackerel, blue whiting, bogue, European anchovy, hake and pilchard and megrim), collected from local commercial fishing vessels were histologically sexed and 5S and 18S rRNA concentrations were quantified by capillary electrophoresis to calculate a 5S/18S rRNA index. Degenerate primers permitted cloning and sequencing of gtf3a fragments in 7 of the studied species. 5S rRNA and gtf3a transcript levels, together with 5S/18S rRNA index, distinguished clearly ovaries from testis in all of the studied species. The values were always higher in females than in males. 5S/18S rRNA index values in females were always highest when fish were captured in early phases of ovary development whilst, in later vitellogenic stages, the values decreased significantly. In megrim and European anchovy, where gonads in different oogenesis stages were obtained, the 5S/18S rRNA index identified clearly gametogenic stage. This approach, to the sexing and the quantitative non-subjective identification of the maturity stage of female fish, could have multiple applications in the study of fish stock dynamics, fish reproduction and fecundity and fish biology in general.

Streptococcus pyogenes Pneumonia in Adults: Clinical Presentation and Molecular Characterization of Isolates 2006-2015

Introduction In the preantibiotic era Streptococcus pyogenes was a common cause of severe pneumonia but currently, except for postinfluenza complications, it is not considered a common cause of community-acquired pneumonia in adults. Aim and Material and Methods This study aimed to identify current clinical episodes of S. pyogenes pneumonia, its relationship with influenza virus circulation and the genotypes of the involved isolates during a decade in a Southern European region (Gipuzkoa, northern Spain). Molecular analysis of isolates included emm, multilocus-sequence typing, and superantigen profile determination. Results Forty episodes were detected (annual incidence 1.1 x 100,000 inhabitants, range 0.29-2.29). Thirty-seven episodes were community-acquired, 21 involved an invasive infection and 10 developed STSS. The associated mortality rate was 20%, with half of the patients dying within 24 hours after admission. Influenza coinfection was confirmed in four patients and suspected in another. The 52.5% of episodes occurred outside the influenza seasonal epidemic. The 67.5% of affected persons were elderly individuals and adults with severe comorbidities, although 13 patients had no comorbidities, 2 of them had a fatal outcome. Eleven clones were identified, the most prevalent being emm1/ST28 (43.6%) causing the most severe cases. Conclusions S. pyogenes pneumonia had a continuous presence frequently unrelated to influenza infection, being rapidly fatal even in previously healthy individuals.