Identificação e caracterização de pequenos RNAs não codificantes e genes alvos envolvidos em estresse abiótico (seca e salinidade) em Eugenia uniflora L. (Myrtaceae)

As plantas, por serem organismos sésseis, enfrentam persistentemente perturbações ambientais adversas denominadas estresses abióticos, sendo as mais importantes, a seca, a salinidade do solo, as temperaturas extremas e a presença de metais pesados. Em resposta, as plantas desenvolveram mecanismos de tolerância, resistência e prevenção para minimizar a influência do estresse, utilizando estratégias de curto prazo para readaptar rápida e eficientemente seu metabolismo. Neste sentido, os pequenos RNAs não codificantes (sncRNAs) são fortes candidatos para realizar este tipo de regulação. Através do sequenciamento de nova geração revelou-se o papel dos sncRNAs na regulação da expressão gênica em nível transcricional e póstranscricional. Dentre os sncRNAs, os microRNAs (miRNAs) são os mais conhecidos e os fragmentos derivados dos RNAs transportadores (tRFs) são os mais novos e com maiores perspectivas de descobertas futuras. Os miRNAs desempenham papéis regulatórios essenciais tanto no crescimento das plantas quanto no desenvolvimento e resposta ao estresse, enquanto os tRFs, em sua maioria, têm sido associados a respostas de estresse. Eugenia uniflora L., “pitanga” ou a cereja brasileira é uma árvore frutífera nativa da América do Sul que pertence à família Myrtaceae. Ela cresce em diferentes ambientes; florestas, restingas e ambientes áridos e semi-áridos no nordeste brasileiro, sendo uma espécie versátil em termos de adaptabilidade e que desempenha um papel fundamental na manutenção da vegetação costeira arbustiva. Além disso, é muito conhecida por suas propriedades medicinais que são atribuídas aos metabólitos especializados presentes nas folhas e frutos. E. uniflora representa uma fonte fascinante da biodiversidade do germoplasma e tem um grande potencial como fonte de genes para o melhoramento genético. Portanto, a compreensão dos mecanismos que conferem tolerância ao estresse nesta planta é de particular importância. Nesse contexto, o objetivo do presente trabalho é a identificação de sncRNAs (miRNAs e tRFs) por ferramentas de bioinformática e análise do padrão de expressão destes sob condições de estresse abiótico (seca e salinidade), bem como avaliação dos genes envolvidos nesta resposta. No capítulo 1, bibliotecas de DNA, pequenos RNAs (sRNAs) e RNAseq de folhas foram usadas para identificar pre-miRNAs, miRNAs maduros e potenciais alvos destes miRNAs, respectivamente. A montagem de novo do genoma permitiu identificar 38 miRNAs conservados e 28 novos miRNAs. Após a avaliação da expressão destes, 11 conservados, entre eles miR156 e miR170, mostraram variação significativa nas condições de restinga e de estresse induzido por PEG. A maioria deles foram previamente descritos em processos de estresse em outras espécies. 14 novos miRNAs foram avaliados em diferentes tecidos de pitanga mostrando variação significativa no padrão de expressão. Os alvos destes últimos miRNAs foram preditos e validados por RTqPCR. Eles correspondem a genes de fatores de transcrição e outros genes como transferases ou ATPases e demonstraram o padrão esperado oposto à expressão dos miRNAs. No capítulo 2, as mesmas bibliotecas foram usadas para identificar tRFs conservados na família das Myrtaceae. Para isso, os tRNAs de Eucalyptus grandis e E. uniflora foram anotados e os tRNAs comuns foram utilizados para o ancoramento dos sRNAs. 479 tRFs foram identificados em pitanga, na maioria com 18 nucleotídeos (nt). Um conjunto de 11 tRFs conservados em ambas espécies, assim como seus alvos, foram avaliados em condições de estresse salino e seca demonstrando diferenças significativas dependendo do tipo de estresse. Os alvos identificados correspondem a genes previamente descritos como envolvidos em estresse salino e seca para outras espécies. O presente trabalho apresenta fortes evidências do envolvimento dos miRNAs em processos de desenvolvimento e estresse, assim como dos tRFs na resposta à seca e estresse salino presente em E. uniflora. Além disso, os dados produzidos poderão ser utilizados em estudos funcionais mais aprofundados que servirão para melhor compreensão dos mecanismos de tolerância presentes nesta importante planta.Plants being sessile organisms, persistently face adverse environmental perturbations termed as abiotic stresses, most important being drought, soil salinity, extreme temperatures, and heavy metals. They developed several strategies such as tolerance, resistance, and avoidance to minimize stress influence, thus require short-term strategies to quickly and efficiently readapt their metabolism. In this sense, small non coding RNAs are strong candidates to do this kind of fine tune regulation. Next generation sequencing technologies have revealed the key role of these sncRNAs in the transcriptional and posttranscriptional gene-expression regulation. Among the myriad of new sncRNAs, miRNAs are the most known ones and the fragments derived from tRNAs (tRFs) are the newest but with high perspective ones. The miRNAs are endogenous small RNAs that play essential regulatory roles in plant growth, development and stress response. In the case of tRFs, they are mainly involved in stress response. Eugenia uniflora L., ‘pitanga’ or Brazilian cherry is a fruit tree native to South America that belongs to Myrtaceae family. It grows in several different harsh environments, including forests, restingas, near the beach, and arid and semiarid environments in the Brazilian northeast. This species is very versatile in terms of adaptability and plays a fundamental role in the maintenance of the shrubby coastal vegetation. However, this species is best-known because its medicinal properties that are attributed to specialized metabolites with known biological activities present in their leaves and fruits. E. uniflora is a fascinating reservoir of germplasm biodiversity and has great potential as a source of genes for plant breeding. Therefore, understanding the mechanisms conferring stress tolerance will be very useful. In this sense, the objective of this work is to identify sncRNAs (miRNAs and tRFs) by bioinformatic tools and to analyze their expression pattern under stress conditions as well as the genes involved in that response. In chapter 1, DNA, small RNA (sRNA) and RNAseq libraries from leaves were used to identify pre-miRNAs, mature miRNAs and potential targets of these miRNAs, respectively. De novo assembly of the genome identified 38 conserved miRNAs and 28 novel miRNAs. After evaluating their expression pattern, 11 11 conserved miRNAs, including miR156 and miR170, showed significant variation in the natural (restinga habitat) and PEG induced stress. Most of them were previously reported in stress processes. 14 novel miRNAs were evaluated in different tissues of pitanga showing significant variation in the expression pattern. The targets of the last miRNAs were predicted and validated by RT-qPCR. They were transcription factor genes and other genes such as transferases or ATPases and showed the expected opposite pattern to miRNA expression. In Chapter 2, the same libraries were used to identify conserved tRFs in the Myrtaceae family. To do this, the tRNAs of Eucalyptus grandis and E. uniflora were annotated and sRNAs mapped into them. 479 tRFs were identified in pitanga with predominance of those with 18 nucleotide length. 11 conserved tRFs in both species, as well as their targets, were evaluated under saline and drought stress conditions showing significant differences depending on the stress type. The targets were genes previously involved in saline and drought stress for other species. The present work shows strong evidences of the involvement of the miRNAs in the development and stress, as well as the tRFs in the tolerance to drought and saline stress of E. uniflora. In addition, the data could be used in more detailed functional studies that will serve to corroborate and better understand the mechanism of tolerance present in this important plant

Influencia de los estilos de liderazgo en la satisfacción laboral de los colaboradores del sector bancario en lima centro durante el 2021

El estudio de los estilos de liderazgo y la satisfacción laboral, a nivel investigativo ha despertado bastante interés en diferentes sectores, debido a la importancia que existe en conocer la relación entre las variables, y cómo éstas afectan en el éxito de las organizaciones, sumado a ello existe gran interés alrededor de la percepción de las personas en cuanto a sus experiencias psicológicas durante la etapa laboral (Aldape, Pedrozo, Castillo & Moguel, 2011). Por un lado, es importante el estudio del liderazgo porque es el proceso de influir en las personas en el camino hacia el logro de algún objetivo o meta (Chiang, Gomez & Salazar, 2014). Por otro lado, la satisfacción laboral es importante por el bienestar de los colaboradores, por la productividad y por el desempeño de los colaboradores que se refleja en los resultados de las organizaciones (Chiang et al., 2014). Asimismo, el sector bancario es un sector importante en el Perú, ya que está ligado al crecimiento económico del país, cumpliendo un rol de intermediación financiera e impulso a las pequeñas, medianas y grandes empresas del país (Superintendencia de Banca y Seguros, [SBS], 2021a). Para la presente investigación se aplicó a 369 colaboradores del sector bancario de Lima Centro, el cuestionario adaptado MLQ de Castro y Martina para medir los estilos de liderazgo y la escala de Satisfacción Laboral SL-SPC de Sonia Palma para medir la satisfacción laboral. Como resultado, se determinó la influencia positiva de los estilos de liderazgo en la satisfacción laboral, siendo el liderazgo transaccional y transformacional los más influyentes en los colaboradores del sector.The study of leadership styles and job satisfaction, at the investigative level, has aroused considerable interest in different sectors, due to the importance of knowing the relationship between variables, and how to affect the organization´s success, in addition there is great interest around the perception of people regarding their psychological experiences during the work stage (Aldape, Pedrozo, Castillo & Moguel, 2011). On one hand, leadership is the process of influencing people to achieve their objectives and goals (Chiang, Gomez & Salazar, 2014). On the other hand, the job satisfaction is important for the employees well-being, for the performance and productivity of the employees, which is reflected on the organizations results (Chiang et al., 2014). Likewise, the banking sector in Peru has evolved positively, this sector is relevant due to is linked to the economic growth of the country, fulfilling a role of financial intermediation and promoting small, medium and large companies in the country (Superintendencia de Banca y Seguros, [SBS], 2021a). For the present research, the adapted questionnaire MLQ by Castro and Martina was applied to 369 employees of the banking sector of Central Lima to measure leadership styles and the SL-SPC Job Satisfaction scale of Sonia Palma to measure job satisfaction. As a result, the positive influence of leadership styles on job satisfaction was determined, with transactional and transformational leadership being the most influential in the employees of the sector.Tesi

Biotecnología ambiental

The various industrial sectors, as well as livestock and agricultural activities, are increasing the production of inputs to meet the demand of the worldwide demographic explosion, making a challenge the clean maintenance of water, soil, and air. Therefore, the search for solutions for a pollutant-free environment without compromising economic development has become extremely important. Thereby, biotechnological studies in order to solve environmental issues have been gaining extensive attention through the coupling of technology procedures to biological systems as sustainable solutions to remediate contaminated areas. In this sense, this review covers topics such as the role of Omics era in microbial environmental biotechnology for pollution control as well as the microbial fuel cell use in energy production. Moreover, phytoremediation and the perspective of applying chemical methods are approached as environmentally friendly tools for the pollutant control to improve remediation processes.Los diversos sectores industriales, así como las actividades ganaderas y agrícolas, están aumentando la producción de insumos para satisfacer la demanda de la explosión demográfica mundial, lo cual dificulta el mantenimiento limpio del agua, el suelo y el aire. Por lo tanto, la búsqueda de soluciones para un medio ambiente libre de contaminantes sin comprometer el desarrollo económico se ha vuelto extremadamente importante. De este modo, los estudios biotecnológicos para resolver problemas ambientales han recibido una gran atención a través del acoplamiento de procedimientos tecnológicos a sistemas biológicos como soluciones sostenibles para remediar áreas contaminadas. En este sentido, esta revisión cubre temas como el papel de la era Ómica en la biotecnología ambiental microbiana para el control de la contaminación, así como el uso de celdas de combustible microbianas en la producción de energía. Además, la fitorremediación y la perspectiva de aplicar métodos químicos se abordan como herramientas ecológicas para el control de contaminantes y mejorar los procesos de remediación

Evaluación del efecto sobre el rendimiento y calidad del fruto de dos métodos de injerto en sandía (Citrullus lanatus Thunb) en costa central de Perú

La sandía (Citrullus lanatus Thunb.) es una planta de la familia de las cucurbitáceas cuyo cultivo se está desarrollando aceleradamente, siendo las zonas costeras del país las más relevantes. En la familia Cucurbitáceas, la técnica de injerto se ha utilizado para mejorar la resistencia a los estreses abióticos, productividad del cultivo y características cualitativas de la fruta. En tal sentido, el presente trabajo evalúa los métodos de injerto por empalme y aproximación en sandía var. Peacok Wr 24 (Citrullus lanatus Thunb) injertada sobre Lagenaria siceraria y su efecto sobre el rendimiento y calidad del fruto. El peso del fruto, número de fruto por planta, firmeza de la pulpa, grados brix y rendimiento fueron evaluados, encontrándose diferencias significativas en el contenido de azúcares medido por grados brix cuando el cultivo fue injertado por aproximación. Si bien hubo un incremento en el rendimiento en los frutos de tercera categoría cuando se usó el injerto por aproximación, este no superó los valores en los frutos de primera categoría. En conclusión, los métodos de injerto no aumentaron el rendimiento y calidad del fruto, pero brindaron mayor vigor a la planta y ayudaron en su tolerancia a enfermedades

Comparative genomics suggests differences related to resistance and virulence between food-isolated Listeria monocytogenes serotypes 1/2a and 4b

Among the four lineages described for Listeria monocytogenes (I, II, III, and IV), lineages I and II harbor the serotypes most closely related to listeriosis in humans. Serotypes 1/2b and 4b are associated with the majority of listeriosis outbreaks, and serotype 1/2a is frequently involved in food and processing plant contamination. As such, the present study utilizes phylogenetic analysis for the aim of determining genomic differences between two L. monocytogenes strains isolated in southern Brazil (serotypes 1/2a and 4b) and known reference strains (L. monocytogenes EGD-e and L. monocytogenes Scott A). The Illumina Miseq platform was used to perform genomic sequencing, and cluster analysis of orthologous groups facilitated the investigation of similarities and differences between the two serotypes studied. In line with previous research, the studied strains of serotypes 1/2a and 4b presented different proteins related to resistance and virulence that may represent adaptations to several conditions during its evolution

Prevalencia de bacterias multidrogoresistentes en un hospital público ubicado en la sierra del Perú

Introduction. Antibiotic resistance is one of the greatest threats to global health. One of the strategies for its control is microbiological surveillance. Objective. To describe the variation of the prevalence of multidrugresistant strains (MDR) of the most frequently isolated bacteria in clinical samples of patients treated at a tertiary care hospital in a high-altitude city in Perú and the factors associated with its isolation. Also, to assess the prevalence of other resistance phenotypes. Results. The general prevalence of MDR strains was 74,1%, observing a downward trend in the annual prevalence of MDR strains in five of the nine bacteria included. The factors associated with MDR strains corresponded to those previously described: male sex, age over 75 years, and hospitalization in intensive care services. In addition, an increase in the annual prevalence of other resistance mechanisms was evidenced. Conclusions. A high prevalence of MDR strains was found in all the bacteria evaluated, associated with previously described factors.Introducción. La resistencia antibiótica es una de las mayores amenazas para la salud global. Una de las estrategias para su control, es la vigilancia microbiológica. Objetivo. Describir la variación de la prevalencia de cepas multidrogoresistentes (MDR) de las bacterias más frecuentemente aisladas en muestras clínicas de pacientes atendidos en un hospital de tercer nivel de una ciudad de altura en el Perú, y determinar los factores asociados a su aislamiento. Además, evaluar la prevalencia de otros fenotipos de resistencia. Métodos. Se realizó un estudio observacional transversal a partir de una cohorte histórica de aislamientos entre los años 2012 y 2019. Resultados. La prevalencia general de cepas MDR fue 74,1%, observándose una tendencia a la disminución de la prevalencia anual de cepas de MDR en cinco de las nueve bacterias analizadas. Los factores asociados a cepas MDR se correspondían con los descritos previamente: sexo masculino, edad mayor a 75 años y hospitalización en servicios de cuidados intensivos. Además, se observó un incremento en la prevalencia de otros fenotipos de resistencia. Conclusión. Se encontró una alta prevalencia de cepas MDR en todas las bacterias evaluadas, asociadas a factores previamente descritos

Novel and Conserved miRNAs Among Brazilian Pine and Other Gymnosperms

The knowledge about plant miRNAs has increased exponentially, with thousands of miRNAs been reported in different plant taxa using high throughput sequencing technologies and bioinformatic tools. Nevertheless, several groups of plants remain unexplored, and the gap of knowledge about conifer miRNAs is considerable. There is no sequence or functional information available on miRNAs in Araucariaceae. This group is represented in Brazil by only one species, Araucaria angustifolia, an endangered species known as Brazilian pine. In the present study, Brazilian pine has its transcriptome explored with respect to small RNAs, representing the first description in a member of the Araucariaceae family. The screening for conserved miRNAs in Brazilian pine revealed 115 sequences of 30 miRNA families. A total of 106 precursors sequences were predicted. Forty one comprised conserved miRNAs from 16 families, whereas 65 were annotated as novel miRNAs. The comparison of Brazilian pine precursors with sRNA libraries of other five conifer species indicates that 9 out 65 novel miRNAs are conserved among gymnosperms, while 56 seems to be specific for Brazilian pine or restricted to Araucariaceae family. Analysis comparing novel Brazilian pine miRNAs precursors and Araucaria cunninghamii RNA-seq data identified seven orthologs between both species. Mature miRNA identified by bioinformatics predictions were validated using stem-loop RT-qPCR assays. The expression pattern of conserved and novel miRNAs was analyzed in five different tissues of 3-month-old Araucaria seedlings. The present study provides insights about the nature and composition of miRNAs in an Araucariaceae species, with valuable information on miRNAs diversity and conservation in this taxon

Direct intranasal application of the solid phase of ImmunoCAP ® increases nasal specific immunoglobulin E detection in local allergic rhinitis patients

The measurement of nasal specific IgE (NsIgE) in local allergic rhinitis (LAR) patients is challenging and shows variability. The objective of this work was to evaluate a minimally-invasive method of direct detection of NsIgE in patients with LAR to Dermatophagoides pteronyssinus (DP) using an automated immunoassay. Methods: Fifty patients participated (LAR, n = 14; allergic rhinitis (AR), n = 20; healthy controls [HC], n = 16). Detection of NsIgE was performed by direct application of the solid phase of a commercialDPImmunoCAP ® test 24 hours a er DP nasal provocation. Results: There was no difference in the median volume of secretion absorbed by the solid phase of the Immuno- CAP test in the 3 studied groups (p = 0.17). According to receiver operating characteristic (ROC) curve analysis. NsIgE 0.1450 was the optimal cutoff point, obtaining in LAR patients 42.86% sensitivity with the highest specificity (100%), and 75% sensitivity and 100% specificity for AR. Conclusion: This study demonstrates the detection of NsIgE to DP in LAR by using a simple, commercial device with high specificity

Influence of the doping level of boron-doped diamond anodes on the removal of penicillin G from urine matrixes

The objective of this study is to understand the influence of the characteristics of boron-doped diamond anodes on the degradation of Penicillin G contained in urine. Therefore, five commercial BDD anodes with different boron doping levels (100 ppm - 8000 ppm) were studied. These electrodes were characterized by cyclic voltammetry, electrochemical impedance spectroscopy, and electrolysis. The boron doping was found to correlate well with the electrochemical properties of the electrodes, and results indicate a different behavior in drug degradation. The improvement in the toxicity and the reduction of the antibiotic effect of urine were the most innovative inputs monitored. For this, the concentration of Penicillin G, the toxicity toward Vibrio fisheri, and the antibiotic effect in Enterococcus faecalis were monitored. The best results were found for the BDD with a boron content of 200 ppm, capable of removing 100% of the antibiotic, reducing toxicity by 90%, and eradicating the antibiotic effect. These results indicate that low doping levels are more efficient for urine removal by anodic oxidation

Improving biodegradability of clopyralid wastes by photoelectrolysis: The role of the anode material

In this work, the removal of the non-readily biodegradable herbicide clopyralid by electrochemical (EC) and photoelectrochemical (PhEC) oxidation with different anode materials were conducted looking to improve not only its oxidation but also its biodegradability. First, in order to find out optimal conditions, it was carried out EC and PhEC degradations in chloride medium, at current densities ranging from 30 to 100 mA cm−2 during 1 h (0.8–2.7 A h L−1), using as anodes MMO-RuO2TiO2, MMO-RuO2IrO2, MMO-IrO2Ta2O5 and boron-doped diamond (BDD). Results show better efficiencies on clopyralid removal for MMO-RuO2IrO2 and BDD anodes at lower current densities. Then, the influence of all anodes on clopyralid transformation was evaluated, extending the electrolysis and photoelectrolysis for 8 h applying 30 mA cm−2 (6.4 A h cm−3). At these conditions, better outcomes are observed for PhEC degradation, where complete pollutant removal is attained for BDD anode and 88.7% for MMO-RuO2IrO2, while COD removal is 47.7% for MMO and 43.1% for BDD anode. Then, short-term biodegradability tests, conducted for EC and PhEC processes, pointed out that MMO-RuO2TiO2 is the most promising anode material, being capable of improving biodegradability in 48.2% and 53% for EC and PhEC degradation, respectively. The toxicity of treated solutions using MMO-RuO2TiO2 and BDD anodes in both EC and PhEC degradation were compared, employing the inhibitory effect in the bioluminescence of marine bacteria Vibrio Fisheri. Toxicity assessments show that toxicity significantly reduces by using the MMO-RuO2TiO2 in NaCl and Na2SO4 medium for both processes. Finally, this study demonstrates that photoelectrolysis with MMO anodes was the most effective strategy in order to increase biodegradability in chloride media, as well as to reduce the toxicity of the treated waste