Shining light on the transcriptome: Molecular regulatory networks leading to a fast-growth phenotype by continuous light in an environmentally sensitive teleost (Atherinopsidae).

Photoperiod can profoundly affect the physiology of teleost fish, including accelerated growth here defined as "fast growth phenotypes". However, molecular regulatory networks (MRNs) and biological processes being affected by continuous illumination and which allow some teleost species evident plasticity to thrive under this condition are not yet clear. Therefore, to provide a broad perspective of such mechanisms, Chirostoma estor fish were raised and sampled for growth under a simulated control (LD) 12 h Light: 12 h Dark or a continuous illumination (LL) 24 h Light: 0 h Dark since fertilization. The experiment lasted 12 weeks after hatching (wah), the time at which fish were sampled for growth, length, and whole-body cortisol levels. Additionally, 3 heads of fish from each treatment were used to perform a de novo transcriptome analysis using Next-Generation Sequencing. Fish in LL developed the fast growth phenotype with significant differences visible at 4 wah and gained 66% more mass by 12 wah than LD fish. Cortisol levels under LL were below basal levels at all times compared to fish in LD, suggesting circadian dysregulation effects. A strong effect of LL was observed in samples with a generalized down-regulation of genes except for Reactive Oxygen Species responses, genome stability, and growth biological processes. To our knowledge, this work is the first study using a transcriptomic approach to understand environmentally sensitive MRNs that mediate phenotypic plasticity in fish submitted to continuous illumination. This study gives new insights into the plasticity mechanisms of teleost fish under constant illumination.

Knowledge Discovery for Higher Education Student Retention Based on Data Mining: Machine Learning Algorithms and Case Study in Chile.

Data mining is employed to extract useful information and to detect patterns from often large data sets, closely related to knowledge discovery in databases and data science. In this investigation, we formulate models based on machine learning algorithms to extract relevant information predicting student retention at various levels, using higher education data and specifying the relevant variables involved in the modeling. Then, we utilize this information to help the process of knowledge discovery. We predict student retention at each of three levels during their first, second, and third years of study, obtaining models with an accuracy that exceeds 80% in all scenarios. These models allow us to adequately predict the level when dropout occurs. Among the machine learning algorithms used in this work are: decision trees, k-nearest neighbors, logistic regression, naive Bayes, random forest, and support vector machines, of which the random forest technique performs the best. We detect that secondary educational score and the community poverty index are important predictive variables, which have not been previously reported in educational studies of this type. The dropout assessment at various levels reported here is valid for higher education institutions around the world with similar conditions to the Chilean case, where dropout rates affect the efficiency of such institutions. Having the ability to predict dropout based on student's data enables these institutions to take preventative measures, avoiding the dropouts. In the case study, balancing the majority and minority classes improves the performance of the algorithms.

High levels of docosahexaenoic acid are present in eight New World silversides (Pisces: Atherinopsidae)

Docosahexaenoic acid (DHA) is the most critical and least available omega-3 fatty acid in the Western human diet. Currently, the source of omega-3 long chain polyunsaturated fatty acids (LC-PUFA) is mainly dependent on wild fisheries, making this resource unsustainable in the foreseeable future. In recent years, a high rate of biosynthesis and accumulation of DHA has been discovered in a freshwater species (Chirostoma estor) belonging to the Atherinopsidae family. Interest in evaluating fatty acid composition in other members of the family has emerged, so this study compiles original data of flesh composition of eight atherinopsid species from freshwater and brackish environments, either wild or cultured. High levels of DHA (16 to 31%) were found in all analyzed members of the family, except in C. grandocule, independently of their habitat or origin. The analyzed species of the Jordani group (C. estor, C. promelas and C. humboldtianum) showed high DHA and low EPA levels (<0.5%) as previously reported for cultured C. estor. The low trophic niche of these atherinopsids and their fatty acid accumulation capabilities are factors that make these species noteworthy candidates for sustainable aquaculture.(AU)

O ácido docosahexaenóico (DHA) é o ácido graxo ômega-3 mais importante e menos disponível na dieta humana ocidental. Atualmente, a fonte de ácidos graxos poliinsaturados de cadeia longa ômega-3 (LC-PUFA) depende principalmente da pesca extrativista, tornando esse recurso insustentável em um futuro próximo. Nos últimos anos, uma alta taxa de biossíntese e acúmulo de DHA foi descoberta em uma espécie de água doce (Chirostoma estor) pertencente à família Atherinopsidae. Deste modo, surgiu o interesse em avaliar a composição de ácidos graxos em outros membros da família. Portanto, este estudo compila dados originais da composição de carne de oito espécies de aterinopsídeos de ambientes de água doce e salobra, selvagens ou cultivadas. Altos níveis de DHA (16 a 31%) foram encontrados em todos os membros da família analisados, exceto em C. grandocule, independentemente de seu habitat ou origem. As espécies analisadas do grupo Jordani (C. estor, C. promelas e C. humboldtianum) apresentaram altos níveis de DHA e EPA baixos (<0,5%), como relatado anteriormente para C. estor cultivado. O baixo nicho trófico desses aterinopsídeos e sua capacidade de acumulação de ácidos graxos são fatores que tornam essas espécies notáveis candidatas à aquicultura sustentável.(AU)

Experimental evidence of masculinization by continuous illumination in a temperature sex determination teleost (Atherinopsidae) model: is oxidative stress involved?

The present study evaluates the influence of continuous light on phenotypic sex ratios in Chirostoma estor, a temperature sex determination animal model. Relative gene expression levels of 5 day old larvae were performed on two early gonad differentiation genes (sox9 and foxl2), two stress axis activation genes (gcr1 and crf) and four reactive oxygen species (ROS) antagonist effector genes (sod2, ucp2, gsr and cat). Two light treatments were applied from fertilization; control (12L:12D) simulated natural photoperiod and a continuous illumination photoperiod. By the end of the trial (12 weeks after hatching), differentiated and normal gonads were clearly identifiable in both treatments by histological observations. Regarding sex ratio, 73% of phenotypic males were found in continuous illumination compared with 40% in controls. Consistently, the sox9 gene (involved in early testis differentiation) showed an over expression in 64% of the individual larvae analysed compared with foxl2 (ovarian differentiation) suggesting a masculinization tendency in continuous illumination. On the other hand, only 36% of individuals showed the same tendency in the control treatment consistent with phenotypic sex ratios found under normal culture conditions. Relative gene expression results did not show significant difference in sod2, ucp2 and gcr1 levels, but cat, gsr and crf showed significantly higher expression levels in the continuous illumination treatment suggesting that both, the stress axis and ROS response mechanisms were activated at this time. This study suggests, a link between continuous light, oxidative stress and environmental sex determination in vertebrates. However, further research is necessary to describe this possible upstream mechanism that may drive some aspects of sexual plasticity in vertebrates.

Evaluation of the immunogenicity of a recombinant HSV-1 vector expressing human group C rotavirus VP6 protein.

Group C Rotavirus (RVC) has been associated globally with sporadic outbreaks of gastroenteritis in children and adults. RVC also infects animals, and interspecies transmission has been reported as well as its zoonotic potential. Considering its genetic diversity and the absence of effective vaccines, it is important and necessary to develop new generation vaccines against RVC for both humans and animals. The aim of the present study was to develop and characterize an HSV-1-based amplicon vector expressing a human RVC-VP6 protein and evaluate the humoral immune response induced after immunizing BALB/c mice. Local fecal samples positive for RVC were used for isolation and sequencing of the vp6 gene, which phylogenetically belongs to the I2 genotype. We show here that cells infected with the HSV[VP6C] amplicon vector efficiently express the VP6 protein, and induced specific anti-RVC antibodies in mice immunized with HSV[VP6C], in a prime-boost schedule. This work highlights that amplicon vectors are an attractive platform for the generation of safe genetic immunogens against RVC, without the addition of external adjuvants.

Impacto de la metodología Lean en la mejora de procesos asistenciales y niveles de satisfacción en la atención de pacientes en un laboratorio clínico / Impact of Lean methodology to improve care processes and levels of satisfaction in patient care in a clinical laboratory

Introducción. La aplicación de la metodología Lean en las instituciones de salud es una herramienta efectiva para mejorar la capacidad y el flujo de trabajo, así como para incrementar el nivel de satisfacción de pacientes y empleados. Objetivo. Optimizar el tiempo de atención de los pacientes ambulatorios en un laboratorio clínico mediante la implementación de una metodología basada en la organización de procesos operativos, para mejorar la satisfacción del usuario y reducir el número de quejas por demoras en la atención. Materiales y métodos. Se implementó un estudio cuasi experimental de antes y después, realizado entre octubre de 2011 a septiembre de 2012. Se emplearon gráficos X Barr y S para observar los promedios en los tiempos de atención y su desviación estándar. La satisfacción de los usuarios se evaluó mediante encuestas de servicio. Resultados. Se observó una disminución de hasta 9 min en los tiempos de atención de los pacientes, desde su llegada hasta su salida del laboratorio, y una disminución del 73% en las quejas por demora en la atención. A pesar de la alta rotación del personal y del incremento del 38% en el número de pacientes atendidos, se adquirió una cultura de empoderamiento y mejora continua, así como mayor eficiencia y productividad en el proceso de atención; lo cual se vio reflejado por el mantenimiento de los estándares 12 meses después de la implementación. Conclusión. La metodología Lean es una herramienta viable para mejorar los procesos de los laboratorios clínicos, mejorando su eficiencia y eficacia (AU)

Introduction. The application of the Lean methodology in health institutions is an effective tool to improve the capacity and workflow, as well as to increase the level of satisfaction of patients and employees. Objective. To optimise the time of outpatient care in a clinical laboratory, by implementing a methodology based on the organisation of operational procedures to improve user satisfaction and reduce the number of complaints for delays in care. Material and methods. A quasi-experimental before and after study was conducted between October 2011 to September 2012. XBar and S charts were used to observe the mean service times and standard deviation. The user satisfaction was assessed using service questionnaires. Results. A reduction of 17 minutes was observed in the time of patient care from arrival to leaving the laboratory, and a decrease of 60% in complaints of delay in care. Despite the high staff turnover and 38% increase in the number of patients seen, a culture of empowerment and continuous improvement was acquired, as well as greater efficiency and productivity in the care process, which was reflected by maintaining standards 12 months after implementation. Conclusion. Lean is a viable methodology for clinical laboratory procedures, improving their efficiency and effectiveness (AU)

[Impact of Lean methodology to improve care processes and levels of satisfaction in patient care in a clinical laboratory]. / Impacto de la metodología Lean en la mejora de procesos asistenciales y niveles de satisfacción en la atención de pacientes en un laboratorio clínico.

INTRODUCTION: The application of the Lean methodology in health institutions is an effective tool to improve the capacity and workflow, as well as to increase the level of satisfaction of patients and employees. OBJECTIVE: To optimise the time of outpatient care in a clinical laboratory, by implementing a methodology based on the organisation of operational procedures to improve user satisfaction and reduce the number of complaints for delays in care. MATERIAL AND METHODS: A quasi-experimental before and after study was conducted between October 2011 to September 2012. XBar and S charts were used to observe the mean service times and standard deviation. The user satisfaction was assessed using service questionnaires. RESULTS: A reduction of 17 minutes was observed in the time of patient care from arrival to leaving the laboratory, and a decrease of 60% in complaints of delay in care. Despite the high staff turnover and 38% increase in the number of patients seen, a culture of empowerment and continuous improvement was acquired, as well as greater efficiency and productivity in the care process, which was reflected by maintaining standards 12 months after implementation. CONCLUSION: Lean is a viable methodology for clinical laboratory procedures, improving their efficiency and effectiveness.

Diversification of substrate specificities in teleostei Fads2: characterization of Δ4 and Δ6Δ5 desaturases of Chirostoma estor.

Currently existing data show that the capability for long-chain PUFA (LC-PUFA) biosynthesis in teleost fish is more diverse than in other vertebrates. Such diversity has been primarily linked to the subfunctionalization that teleostei fatty acyl desaturase (Fads)2 desaturases have undergone during evolution. We previously showed that Chirostoma estor, one of the few representatives of freshwater atherinopsids, had the ability for LC-PUFA biosynthesis from C18 PUFA precursors, in agreement with this species having unusually high contents of DHA. The particular ancestry and pattern of LC-PUFA biosynthesis activity of C. estor make this species an excellent model for study to gain further insight into LC-PUFA biosynthetic abilities among teleosts. The present study aimed to characterize cDNA sequences encoding fatty acyl elongases and desaturases, key genes involved in the LC-PUFA biosynthesis. Results show that C. estor expresses an elongase of very long-chain FA (Elovl)5 elongase and two Fads2 desaturases displaying Δ4 and Δ6/Δ5 specificities, thus allowing us to conclude that these three genes cover all the enzymatic abilities required for LC-PUFA biosynthesis from C18 PUFA. In addition, the specificities of the C. estor Fads2 enabled us to propose potential evolutionary patterns and mechanisms for subfunctionalization of Fads2 among fish lineages.

A complete approach for recombinant protein expression training: From gene cloning to assessment of protein functionality*.

A practical course was given to undergraduate biology students enrolled in the elective course "Introduction to Genetic Engineering" at the Federal University of São Carlos (UFSCar), São Paulo, Brazil. The goal of the course was to teach current molecular biology tools applied to a real research situation that could be reported by the students themselves. The purpose was to produce a plant recombinant protein and demonstrate a heretofore unreported biological activity. Cystatins, natural inhibitors of cysteine proteases, were proposed for these studies. Initially, the students searched for plant cystatin cDNA sequences in the NCBI databases and selected the Oryzacystatin I gene (ocI) from rice, Oriza sativa, as the target gene for this study. Total RNA was extracted from rice-germinating seeds and primers containing restriction sites for NdeI and EcoRI were designed based on the ocI cDNA sequence and then used to amplify the open reading frame (ORF). RT-PCR amplification provided a band of the expected size for ocI ORF (309 bp). The PCR product was cut with NdeI and EcoRI restriction enzymes and cloned directly in the pET28a expression vector digested with the same enzymes. A pET28-ocI recombinant clone was selected, checked by sequencing, and used to transform Escherichia coli BL21 (DE3) expression strain. After induction of the bacteria with isopropylthiogalactoside and cellular disruption, the His-tagged OCI protein, present mainly in the soluble fraction, was purified by affinity chromatography in a nickel column. The purified protein was successfully used to inhibit fungal growth (Trichoderma reesei). The results were discussed extensively and the students contributed to the writing of this article, of which they are co-authors.