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Recent advances in high-throughput molecular methods have led to an extraordinary volume of genomics data. Simultaneously, the progress in the computational implementation of novel algorithms has facilitated the creation of hundreds of freely available online tools for their advanced analyses. However, a general overview of the most commonly used tools for the in silico analysis of genomics data is still missing. In the current article, we present an overview of commonly used online resources for genomics research, including over 50 tools. This selection will be helpful for scientists with basic or intermediate skills in the in silico analyses of genomics data, such as researchers and students from wet labs seeking to strengthen their computational competencies. In addition, we discuss current needs and future perspectives within this field.

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Malignant pleural mesothelioma (MPM) is a rare and aggressive neoplasm of the pleural tissue that lines the lungs and is mainly associated with long latency from asbestos exposure. This tumor has no effective therapeutic opportunities nowadays and has a very low five-year survival rate. In this sense, identifying molecular events that trigger the development and progression of this tumor is highly important to establish new and potentially effective treatments. We conducted a meta-analysis of genome-wide expression studies publicly available at the Gene Expression Omnibus (GEO) and ArrayExpress databases. The differentially expressed genes (DEGs) were identified, and we performed functional enrichment analysis and protein-protein interaction networks (PPINs) to gain insight into the biological mechanisms underlying these genes. Additionally, we constructed survival prediction models for selected DEGs and predicted the minimum drug inhibition concentration of anticancer drugs for MPM. In total, 115 MPM tumor transcriptomes and 26 pleural tissue controls were analyzed. We identified 1046 upregulated DEGs in the MPM samples. Cellular signaling categories in tumor samples were associated with the TNF, PI3K-Akt, and AMPK pathways. The inflammatory response, regulation of cell migration, and regulation of angiogenesis were overrepresented biological processes. Expression of SOX17 and TACC1 were associated with reduced survival rates. This meta-analysis identified a list of DEGs in MPM tumors, cancer-related signaling pathways, and biological processes that were overrepresented in MPM samples. Some therapeutic targets to treat MPM are suggested, and the prognostic potential of key genes is shown.

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Social cognition impairments may be associated with poor functional outcomes, symptoms, and disability in social anxiety disorder (SAD) and generalized anxiety disorder (GAD). This meta-analysis aims to determine if emotion recognition and theory of mind (ToM) are impaired in SAD or GAD compared to healthy controls. A systematic review was conducted in electronic databases (PubMed, PsycNet, and Web of Science) to retrieve studies assessing emotion recognition and/or ToM in patients with SAD or GAD, compared to healthy controls, up to March 2022. Meta-analyses using random-effects models were conducted. We identified 21 eligible studies: 13 reported emotion recognition and 10 ToM outcomes, with 585 SAD patients, 178 GAD patients, and 753 controls. Compared to controls, patients with SAD exhibited impairments in emotion recognition (SMD = -0.32, CI = -0.47 - -0.16, z = -3.97, p < 0.0001) and ToM (SMD = -0.44, CI = -0.83 -0.04, z = -2.18, p < 0.01). Results for GAD were inconclusive due to the limited number of studies meeting the inclusion criteria (two for each domain). Relevant demographic and clinical variables (age, sex, education level, and anxiety scores) were not significantly correlated with emotion recognition or ToM impairments in SAD and GAD. Further studies employing ecological measures with larger and homogenous samples are needed to better delineate the factors influencing social cognition outcomes in both SAD and GAD.

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In recent decades, advances in methods in molecular biology and genetics have revolutionized multiple areas of the life and health sciences. However, there remains a global need for the development of more refined and effective methods across these fields of research. In this current Collection, we aim to showcase articles presenting novel molecular biology and genetics techniques developed by scientists from around the world.

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Abstract Introduction: The SARS-CoV-2 is the denomination of the new betacoronavirus, which was discovered and isolated for the first time in Wuhan, China, at the end of December 2019, and it is the causal agent of the sanitary emergency of the COVID-19 pandemic. Experimental studies have shown susceptibility to infection in pets (dogs and cats). Objective: To present the current information available on SARS-CoV-2 in animals under the care of humans that have been officially reported in the sanitary registries of the World Animal Health Information System (WAHIS) of the World Organization for Animal Health. Materials and methods: We conducted a narrative review using Medline/ PubMed, Scopus, and Web of Sciences, and official documents of the World Organisation for Animal Health. The search terms used were as follows: "coronavirus", "SARS coronavirus 2019", "SARS-CoV", "SARS-CoV-2 in dog and/or cat" "pets SARS-CoV-2". Results: The studies reviewed in this manuscript highlight those positive cases in cats and dogs for SARS-CoV-2 have been associated with an exposure to positive COVID-19 people. In the available evidence, 55.17 % of the total cases of animals that were positive for SARS-CoV-2 were associated with people with COVID-19 who had the disease at home, possibly due to maintaining a longer exposure to the humans. Conclusion: Regarding the zoonotic aspects, it is important to clarify that although several animal species have been infected by SARS-CoV-2, none of them has been scientifically proven to represent a risk of direct transmission between positive animals and other humans or to play an epidemiological role in the disease.

Resumen Introducción: El SARS-CoV-2 es el nombre para el nuevo betacoronavirus, que fue descubierto y aislado por primera vez en Wuhan, China, a fines de diciembre de 2019, y es el agente causal de la emergencia sanitaria del COVID-19. Estudios experimentales han demostrado susceptibilidad a la infección en mascotas (perros y gatos). Objetivo: Presentar la información actual sobre el SARS-CoV-2 en animales bajo el cuidado de humanos que han sido oficialmente reportados en los registros sanitarios del Sistema Mundial de Información Sanitaria Animal (WAHIS) de la Organización Mundial de Sanidad Animal. Materiales y métodos: Se realizó una revisión narrativa utilizando Medline/PubMed, Scopus y Web of Sciences, y documentos oficiales de la Organización Mundial de Sanidad Animal. Los términos de búsqueda utilizados fueron los siguientes: "coronavirus", "SARS coronavirus 2019", "SARS-CoV", "SARS-CoV-2 en perro y/o gato" "mascotas SARS-CoV-2". Resultados: Los estudios revisados en este manuscrito destacan que los casos positivos en gatos y perros para SARS-CoV-2 se han asociado con una exposición a personas positivas para COVID-19. En la evidencia disponible, el 55,17 % del total de casos de animales positivos para SARS-CoV-2 estaban asociados a personas con COVID-19 que tenían la enfermedad en casa, posiblemente por mantener una mayor exposición a los humanos. Conclusión: En cuanto a los aspectos zoonóticos, es importante aclarar que si bien varias especies animales han sido infectadas por el SARS-CoV-2, ninguna de ellas ha demostrado científicamente que represente un riesgo de transmisión directa entre animales positivos y otros humanos o que juegue un papel epidemiológico en la enfermedad.

Resumo Introdução: SARS-CoV-2 é o nome do novo betacoronavírus, que foi descoberto e isolado pela primeira vez em Wuhan, China, no final de dezembro de 2019, e é o agente causal da emergência sanitária COVID-19. Estudos experimentais mostraram suscetibilidade à infecção em animais de estimação (cães e gatos). Objetivo: Apresentar as informações atuais sobre SARS-CoV-2 em animais sob cuidados humanos que foram oficialmente notificados nos registros sanitários do World Animal Health Information System (WAHIS) da Organização Mundial de Saúde Animal. Materiais e métodos: Foi realizada uma revisão narrativa utilizando Medline/PubMed, Scopus e Web of Sciences e documentos oficiais da Organização Mundial de Saúde Animal. Os termos de pesquisa utilizados foram os seguintes: "coronavírus", "SARS coronavirus 2019", "SARS-CoV", "SARS-CoV-2 in dogs and/ or cats" "SARS-CoV-2 pets". Resultados: Os estudos revisados neste manuscrito destacam que casos positivos em gatos e cães para SARS-CoV-2 foram associados à exposição a pessoas positivas para COVID-19. Nas evidências disponíveis, 55,17 % do total de casos animais positivos para SARS-CoV-2 foram associados a pessoas com COVID-19 que tiveram a doença em casa, possivelmente devido à maior exposição a humanos. Conclusão: Em relação aos aspectos zoonóticos, é importante esclarecer que, embora várias espécies animais tenham sido infectadas pelo SARS-CoV-2, nenhuma de las foi cientificamente comprovada como representando risco de transmissão direta entre animais positivos e outros humanos ou que desempenhe um papel papel epidemiológico da doença.

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The immune function is closely related to iron (Fe) homeostasis and allostasis. The aim of this bioinformatics-assisted review was twofold; (i) to update the current knowledge of Fe metabolism and its relationship to the immune system, and (ii) to perform a prediction analysis of regulatory network hubs that might serve as potential biomarkers during stress-induced immunosuppression. Several literature and bioinformatics databases/repositories were utilized to review Fe metabolism and complement the molecular description of prioritized proteins. The Search Tool for the Retrieval of Interacting Genes (STRING) was used to build a protein-protein interactions network for subsequent network topology analysis. Importantly, Fe is a sensitive double-edged sword where two extremes of its nutritional status may have harmful effects on innate and adaptive immunity. We identified clearly connected important hubs that belong to two clusters: (i) presentation of peptide antigens to the immune system with the involvement of redox reactions of Fe, heme, and Fe trafficking/transport; and (ii) ubiquitination, endocytosis, and degradation processes of proteins related to Fe metabolism in immune cells (e.g., macrophages). The identified potential biomarkers were in agreement with the current experimental evidence, are included in several immunological/biomarkers databases, and/or are emerging genetic markers for different stressful conditions. Although further validation is warranted, this hybrid method (human-machine collaboration) to extract meaningful biological applications using available data in literature and bioinformatics tools should be highlighted.

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There has been an important global interest in Open Science, which include open data and methods, in addition to open access publications. It has been proposed that public availability of raw data increases the value and the possibility of confirmation of scientific findings, in addition to the potential of reducing research waste. Availability of raw data in open repositories facilitates the adequate development of meta-analysis and the cumulative evaluation of evidence for specific topics. In this commentary, we discuss key elements about data sharing in open repositories and we invite researchers around the world to deposit their data in them.

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Creatine (Cr) and phosphocreatine (PCr) are physiologically essential molecules for life, given they serve as rapid and localized support of energy- and mechanical-dependent processes. This evolutionary advantage is based on the action of creatine kinase (CK) isozymes that connect places of ATP synthesis with sites of ATP consumption (the CK/PCr system). Supplementation with creatine monohydrate (CrM) can enhance this system, resulting in well-known ergogenic effects and potential health or therapeutic benefits. In spite of our vast knowledge about these molecules, no integrative analysis of molecular mechanisms under a systems biology approach has been performed to date; thus, we aimed to perform for the first time a convergent functional genomics analysis to identify biological regulators mediating the effects of Cr supplementation in health and disease. A total of 35 differentially expressed genes were analyzed. We identified top-ranked pathways and biological processes mediating the effects of Cr supplementation. The impact of CrM on miRNAs merits more research. We also cautiously suggest two dose-response functional pathways (kinase- and ubiquitin-driven) for the regulation of the Cr uptake. Our functional enrichment analysis, the knowledge-based pathway reconstruction, and the identification of hub nodes provide meaningful information for future studies. This work contributes to a better understanding of the well-reported benefits of Cr in sports and its potential in health and disease conditions, although further clinical research is needed to validate the proposed mechanisms.

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Creatine (Cr) is a ubiquitous molecule that is synthesized mainly in the liver, kidneys, and pancreas. Most of the Cr pool is found in tissues with high-energy demands. Cr enters target cells through a specific symporter called Na+/Cl--dependent Cr transporter (CRT). Once within cells, creatine kinase (CK) catalyzes the reversible transphosphorylation reaction between [Mg2+:ATP4-]2- and Cr to produce phosphocreatine (PCr) and [Mg2+:ADP3-]-. We aimed to perform a comprehensive and bioinformatics-assisted review of the most recent research findings regarding Cr metabolism. Specifically, several public databases, repositories, and bioinformatics tools were utilized for this endeavor. Topics of biological complexity ranging from structural biology to cellular dynamics were addressed herein. In this sense, we sought to address certain pre-specified questions including: (i) What happens when creatine is transported into cells? (ii) How is the CK/PCr system involved in cellular bioenergetics? (iii) How is the CK/PCr system compartmentalized throughout the cell? (iv) What is the role of creatine amongst different tissues? and (v) What is the basis of creatine transport? Under the cellular allostasis paradigm, the CK/PCr system is physiologically essential for life (cell survival, growth, proliferation, differentiation, and migration/motility) by providing an evolutionary advantage for rapid, local, and temporal support of energy- and mechanical-dependent processes. Thus, we suggest the CK/PCr system acts as a dynamic biosensor based on chemo-mechanical energy transduction, which might explain why dysregulation in Cr metabolism contributes to a wide range of diseases besides the mitigating effect that Cr supplementation may have in some of these disease states.

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It has been estimated that epilepsies are among the top five neurological diseases with the highest burden of disease. In recent years, genome-wide expression studies (GWES) have been carried out in experimental models of epilepsy and in samples from human patients. In this study, I carried out meta-analyses and analyses of convergence for available GWES for epileptogenesis in humans and in mouse, rat, zebrafish and fruit fly models. Multiple lines of evidence (such as genome-wide association data and known druggable genes) were integrated to prioritize top candidate genes for epileptogenesis and a functional enrichment analysis was carried out. Several top candidate genes, which are supported by multiple lines of genomic evidence, such as GRIN1, KCNAB1 and STX1B, were identified. Druggable genes of potential interest (such as GABRA2, GRIK1, KCNAB1 and STX4) were also identified. An enrichment of genes regulated by the MEF2 and SOX5 transcription factors and the miR-106b-5p and miR-101-3p miRNAs was found. The current work is the first meta-analysis and convergent analysis of GWES for epileptogenesis in humans and in multiple animal models, integrating results from several genomic studies. Novel candidate genes and pathways for epileptogenesis were identified in this analysis.

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Publishing articles in international scientific journals is the primary method for the communication of validated research findings and ideas. Journal articles are commonly used as a major input for evaluations of researchers and institutions. Few articles have been published previously about the different aspects needed for writing high-quality articles. In this manuscript, we provide an updated and brief guide for the multiple dimensions needed for writing manuscripts in the health and biological sciences, from current, international and interdisciplinary perspectives and from our expertise as authors, peer reviewers and editors. We provide key suggestions for writing major sections of the manuscript (e.g. title, abstract, introduction, methods, results and discussion), for submitting the manuscript and bring an overview of the peer review process and  of the post-publication impact of the articles.

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Resilience is a mechanism used by humans to adapt to adverse situations. It is a protective factor against mental health problems. This process can be influenced by environmental and genetic factors. Several genes have been associated with interindividual differences in resilience levels, but the results are inconclusive. Therefore, the aim of this meta-analysis was to evaluate the effect of a functional polymorphism (5-HTTLPR) in the SLC6A4 gene on resilience levels. A search in PubMed, HugeNavigator and Google Scholar databases was carried out and 16 studies about the association of 5-HTTLPR polymorphism and resilience in humans were identified. The OpenMeta[Analyst] program was employed to perform statistical analysis using a random-effects model. The final analysis included 9 studies, for a total of 4,080 subjects. Significant results were found when the standardized mean differences (SMD) of LL and SL carriers were compared, (SMD: -0.087 (confidence interval: -0.166 to -0.008; I 2 : 0 %); P value: 0.031). A significant result was also found in an analysis comparing SS/SL versus LL genotypes (SMD: -0.231; confidence interval: -0.400 to -0.061, P value: 0.008; I 2 : 0 %). This is the first meta-analysis performed to identify the pooled association of a functional polymorphism in the serotonin transporter gene and resilience. The current results suggest that the L/L genotype is associated with resilience. Further studies are necessary to elucidate the role of genetics on the resilience mechanisms.

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Schizophrenia (SZ) is a complex and severe psychiatric disorder, which has a global lifetime prevalence of 0.4% and a heritability of around 0.81. A number of epigenome-wide association studies (EWAS) have been carried out for SZ, with discordant results. The main aim of this study was to carry out an integrative in silico analysis of available genome-wide DNA methylation profiles in schizophrenia. In this work, an integration of multiple lines of evidence (top candidate genes from several EWAS and genome-wide expression and association data) was carried out, in order to identify top differentially methylated (DM) genes for SZ. In addition, functional enrichment and protein-protein interaction analyses were carried out. Several top differentially methylated genes, such as APC, CACNB2, and PRKN, were found, and an enrichment of binding sites for brain-expressed transcription factors, such as FOXO1, MYB, and ZIC3, was also observed. Moreover, a protein-protein interaction network showed a central role for DISC1 and ZNF688 genes, and experimentally validated targets of MIR-137, such as and KCNB2, NRXN1, and SYN2, were identified among DM genes. This is the first integrative in silico analysis of available genome-wide DNA methylation profiles in schizophrenia. This work identified novel candidate genes and pathways for SZ and provides the basis to explore their role in the pathogenesis of SZ in future studies.

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The aim of the current study was to test the hypothesis that a functional polymorphism in the synaptosome associated protein 25 (SNAP25) gene could be associated with impulsivity scores in a sample of young Colombian subjects. Impulsivity has been postulated as an endophenotype for several psychiatric disorders of high epidemiological relevance. There is a need for the study of additional candidate genes for impulsivity. One hundred seventy-five young Colombian subjects completed the Spanish version of the short form of the Barratt Impulsiveness Scale (BIS-15S). A TaqMan assay was used to genotype a functional polymorphism (rs3746544) in the SNAP25 gene. A significant association was found between the functional polymorphism in the SNAP25 gene and impulsivity in the Colombian sample, with subjects carrying T/T and G/G genotypes showing lower mean scores in the non-planning subfactor (p = 0.02). This is the first report of an association of a functional polymorphism in the SNAP25 gene and a subfactor of the BIS-15S scale of impulsivity. In addition, this the first genetic study of impulsivity scores in a Latin American sample. Future studies should explore additional variants in brain-expressed miRNAs and in their binding sites as candidates for impulsivity in different populations.

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Background: Neurosciences research has increased significantly in recent years around the world. It has led to the development of interdisciplinary work, moving from activities from isolated fields (such as biology, psychology or neurology) to research that involves different scientific perspectives. In developing regions, such as Latin America, it has additional challenges, related to available funding and infrastructure.Aim: To analyze key factors in scientific productivity in neurosciences in Latin America.Methods: A bibliometric analysis of the scientific productivity in neurosciences in main five Latin American countries (Argentina, Brazil, Chile, Colombia and Mexico) was carried out.Results: Brazil was the largest producer of scientific articles, and receptor of citations, in neurosciences in 1998-2017, followed by Mexico. We identified highly cited papers, top institutions, networks of authors, main journals and key areas in neurosciences for this period in the 5 countries.Conclusions: Scientific productivity in neurosciences in Latin America would benefit from the consolidation of more regional, interdisciplinary and international research networks. In this work, we discuss key elements for the consolidation of neurosciences research in Latin America.

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Background: Data from the Global Burden of Disease Study 2016 recently estimated that after opioid and cannabis use disorders, cocaine use disorders were among the most common, with around 5.8 million cases around the world. Several genome-wide expression studies (GWES) for cocaine misuse have been carried out in brain tissues from patients and controls and in mouse and rat models.Objectives: In the current work, we used a convergent functional genomics approach to identify novel candidate genes and pathways for cocaine misuse.Methods: We carried out meta-analyses for available GWES for cocaine misuse in humans and mouse and rat models (three, four, and two GWES, respectively). Multiple lines of evidence (GWES, genome-wide association and epigenomic data) were integrated to prioritize top candidate genes, and a functional enrichment analysis was carried out.Results: Several top candidate genes supported by multiple lines of genomic evidence, and with known roles in brain plasticity, were identified: APP, GRIN2A, GRIN2B, KCNA2, MAP4, PCDH10, PPP3CA, SNCB, and SV2C. An enrichment of genes regulated by the AP1 transcription factor was found.Conclusion: This is the first meta-analysis of GWES for cocaine misuse in humans and mouse and rat models. The analysis of convergence of multiple lines of genome-wide evidence identified novel candidate genes and pathways for cocaine misuse, which are of basic and clinical importance.

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Colombia is the second largest country in South America. In this article, we provide an overview of medical education in Colombia, including a description of existing public and private medical schools and available undergraduate and postgraduate programs. Medical education in Colombia has evolved through time, following international trends. In addition to 61 undergraduate medical programs, there are 529 postgraduate clinical, 30 PhD, and 131 Master programs in health sciences in Colombia. We identify current challenges and highlight future perspectives for medical education in Colombia.

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Resumen Introducción: Este artículo busca debatir y problematizar la apropiación indígena de normatividades expedidas por autoridades estatales colombianas acerca del manejo de las medicinas tradicionales indígenas y los medicamentos de la biomedicina en el país, en específico en las comunidades inga y kamsá. Desarrollo: Dicha problemática se comprende de mejor manera cuando se entiende la tradición comercial y la itinerancia de estas comunidades indígenas. Este artículo muestra cómo la normatividad repercute en la forma de percibir el bienestar, los métodos de tratamiento y la enfermedad por parte de la medicina indígena del Putumayo, inserta en medianas y pequeñas poblaciones urbanas del altiplano cundiboyacense de Colombia. Tal situación genera diferentes estrategias de inserción de la medicina indígena por parte de esta población, como las medicinas empaquetadas, semejantes en apariencia a los medicamentos de la biomedicina, estrategia inevitable en el marco del pluralismo médico que necesariamente se presenta en contextos no cerrados, lo que hace que se fortalezcan los estereotipos de las poblaciones indígenas en las ciudades y, por otro lado, la obligatoria comercialización que sufren las medicinas indígenas al momento de salir al público urbano, que no se intercambian, sino que se compran y venden con dinero. Conclusiones: Las normas, leyes y decretos para la regulación de los medicamentos -y las normas que reconocen la medicina tradicional indígena bajo el marco de reconocimiento multicultural- se están creando bajo estereotipos estáticos de 'lo indígena', lo que refuerza la necesidad de unas estrategias de inserción de la medicina indígena en las ciudades por parte de estas comunidades.

Abstract Introduction: This article wants to discuss the indigenous appropriation of regulation, laws, and decrees established by Colombian state authorities about the production of medical drugs and manage of traditional indigenous medicine, specifically in the Inga and Kamsá communities. Development: This social issue it is better understood by the commercial tradition and the regular traveling of those indigenous communities. The article shows how through the normativity is reflected in the way of perceiving welfare, treatment methods, and disease from the indigenous medicine of the Putumayo inserted in medium and small urban populations of the cundiboyacense plateau of Colombia. This situation makes the indigenous population generate strategies of insertion in small towns. Strategies like packaged medicines looking similar in appearance to the medicines of biomedicine, an inevitable strategy in the social frame of medical pluralism that necessarily presents itself in non-closed contexts, that helps to strengthen the stereotypes of indigenous people. On the other hand, the mandatory commercialization that the indigenous medicines suffer. when they go out to the urban public, a context that does not exchange but buys and sells with money. Conclusions: The norms, laws, and decrees for drug regulation -and the norms that recognize traditional indigenous medicine under the framework of multicultural acknowledgment- are being created under static stereotypes of indigenous stereotypes, reinforcing the need for integration strategies of indigenous medicine in the cities by these communities.

Resumo Introdução: Este artigo busca debater e problematizar a apropriação indígena de normatividades expedida por autoridades estatais colombianas acerca da gestão dos medicamentos tradicionais indígenas e os medicamentos da biomedicina no país, especificamente nas comunidades inga e kamsá. Desenvolvimento: Dita problemática compreende-se de melhor maneira quando se entende a tradição comercial e itinerância destas comunidades indígenas. Este artigo mostra como a normatividade repercute na forma de perceber o bem-estar, os métodos de tratamento e a doença por parte da medicina indígena de Putumayo, insere em médias e pequenas populações urbanas do altiplano de Cundinamarca e Boyacá da Colômbia. Tal situação gera diferentes estratégias de inserção da medicina indígena por parte desta população, como os medicamentos empacotados, semelhantes em aparência aos medicamentos da biomedicina, estratégia inevitável no marco do pluralismo médico que necessariamente se apresenta em contextos não fechados, o que faz que se fortalezam os estereótipos das populações indígenas nas cidades e, por outro lado, a obrigatória comercialização que sofrem os medicamentos indígenas ao momento de sair ao público urbano, que não se intercambiam, senão que se compram e se vendem com dinheiro. Conclusões: As normas, leis e decretos para a regulação dos medicamentos -e as normas que reconhecem a medicina tradicional indígena sob o marco de reconhecimento multicultural- estão sendo criadas sob estereótipos estáticos 'do indígena', o que reforça a necessidade de umas estratégias de inserção da medicina indígena nas cidades por parte essas comunidades.

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Several approaches for miRNA expression analysis have been developed in recent years. In this article, we provide an updated and comprehensive review of available qPCR-based methods for miRNA expression analysis and discuss their advantages and disadvantages. Existing techniques involve the use of stem-loop reverse transcriptase-PCR, polyadenylation of RNAs, ligation of adapters or RT with complex primers, using universal or miRNA-specific qPCR primers and/or probes. Many of these methods are oriented towards the expression analysis of mature miRNAs and few are designed for the study of pre-miRNAs and pri-miRNAs. We also discuss findings from articles that compare results from existing methods. Finally, we suggest key points for the improvement of available techniques and for the future development of additional methods.

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