A study of neural-related microRNAs in the developing amphioxus

<p>Abstract</p> <p>Background</p> <p>MicroRNAs are small noncoding RNAs regulating expression of protein coding genes at post-transcriptional level and controlling several biological processes. At present microRNAs have been identified in various metazoans and seem also to be involved in brain development, neuronal differentiation and subtypes specification. An approach to better understand the role of microRNAs in animal gene expression is to determine temporal and tissue-specific expression patterns of microRNAs in different model organisms. Therefore, we have investigated the expression of six neural related microRNAs in amphioxus, an organism having an important phylogenetic position in terms of understanding the origin and evolution of chordates.</p> <p>Results</p> <p>In amphioxus, all the microRNAs we examined are expressed in specific regions of the CNS, and some of them are correlated with specific cell types. In addition, miR-7, miR-137 and miR-184 are also expressed in endodermal and mesodermal tissues. Several potential targets expressed in the nervous system of amphioxus have been identified by computational prediction and some of them are coexpressed with one or more miRNAs.</p> <p>Conclusion</p> <p>We identified six miRNAs that are expressed in the nervous system of amphioxus in a variety of patterns. miR-124 is found in both differentiating and mature neurons, miR-9 in differentiated neurons, miR-7, miR-137 and miR-184 in restricted CNS regions, and miR-183 in cells of sensory organs. Therefore, such amphioxus miRNAs may play important roles in regional patterning and/or specification of neuronal cell types.</p

Caracterizacion molecular del microambiente de la piel en pacientes con vitiligo, para la búsqueda de biomarcadores indicadores de respuesta a tratamiento.

El vitiligo pertenece a una familia de enfermedades cutáneas congénitas o adquiridas, caracterizadas por falta de pigmentación en la piel en forma de máculas bien definidas, acrómicas o hipocrómicas, en las cuales se ha observado el deterioro de la función del melanocito, la célula de la piel encargada de la producción de la melanina. Aunque se ha descrito que esta enfermedad afecta entre el 0.1 y el 2% de la población mundial, su prevalencia varía considerablemente entre la población o grupo étnico analizado. Las causas del vitiligo son complejas y aún no se han comprendido en su totalidad. Varias hipótesis han sido desarrolladas para explicar el proceso de despigmentación, o incluso la destrucción del melanocito. Sin embargo, no dan cuenta del completo espectro de esta enfermedad. Los estudios realizados a la fecha se han enfocado en estudiar la importancia del melanocito como productor de pigmento y describir cómo las alteraciones en el mismo tienen como consecuencia el desarrollo del vitiligo. Sin embargo, en su mayoría estos han sido realizados a través de métodos de cultivo celular, lo que puede alterar los patrones de expresión reales de la célula, en comparación con los presentes en su tejido de origen, por lo que el estudio del microambiente de la piel y el análisis del estado funcional de sus componentes celulares, pueden ayudar a determinar la mejor alternativa terapéutica en la repigmentación. Objetivo: Identificar en la piel de pacientes con vitiligo antes y después de ser sometidos a tratamiento con fototerapia UVB-nb, cambios en patrones de expresión de genes con utilidad pronóstica. Material y Método: Estudio comparativo, prospectivo, ciego con respecto al evaluador, experimental y longitudinal. Una vez obtenido el consentimiento informado, fueron incluidos cuarenta y cinco sujetos con Vitiligo Vulgar (21 sexo Femenino y 24 sexo Masculino; 23 con forma activa del vitiligo vulgar y 22 con su forma estable), provenientes del Estado de Nuevo León, México, que asistieron al Servicio de Dermatología del Hospital Universitario José Eleuterio González (HUJEG), de la Universidad Autónoma de Nuevo León (UANL), Monterrey (MTY), Nuevo León, México. Previo al tratamiento se obtuvieron muestras de sangre para realizar pruebas bioquímicas en el Laboratorio Central y en el Servicio de Endocrinología del Hospital Universitario UANL. Además, se tomaron 2 biopsias de piel en sacabocado de 4 mm en cada paciente, correspondientes a piel lesionada y piel sana respectivamente. Posteriormente fue iniciado el tratamiento utilizado el equipo de fototerapia UVB-nb en cada paciente, 2 veces por semana, 48 sesiones totales. Al finalizar el protocolo se tomaron biopsias controles, una en tejido repigmentado y otra en el tejido que no experimentó respuesta al tratamiento. Las biopsias de piel fueron utilizadas para extraer ARNm mediante RNaeasy MiniKit de QIAGEN, El cual fue cuantificado y utilizado para realizar el análisis de expresión mediante RNA-Seq (análisis de expresión mediante secuenciación masiva de ARN), en genes participantes en las rutas de pigmentación de la piel, apoptosis, estrés oxidativo, traducción de señal y sobrevida celular, con la finalidad de identificar cambios asociados con el estado de la piel del paciente con utilidad pronóstica de tratamiento. Los análisis moleculares y perfiles de expresión de la biopsias de piel fueron realizados en la Unidad de Biología Molecular, Genómica y Secuenciación (UBMGyS) del Centro de Investigación y Desarrollo en Ciencias de la Salud (CIDICS) UANL, MTY, México. Los resultados clínicos de respuesta a tratamiento fueron evaluados por 2 investigadores independientes y cegados a los resultados de la secuenciación masiva de RNA mediante TruSeq de las muestras de piel. Finalmente, el análisis estadístico de los niveles de expresión y su relación con los parámetros clínicos, bioquímicos y respuesta a tratamiento, fue realizado mediante R de Bioconductor, SPSS y BaseSpace de Illumina Resultados: Se observó que la mayoría de los pacientes desarrollan la enfermedad antes de los 30 años, y que el desarrollo de la patología tenía relación con antecedentes heredofamiliares de la patología u otras de naturaleza inmune. La correlación entre los resultados clínicos, bioquímicos y de expresión permitió identificar que bajos niveles de TSH, y un elevado peso en los pacientes puede conducir a una pobre respuesta al tratamiento UVB-nb. Por otra parte, el análisis de expresión permitió detectar diferencias significativas en los patrones de expresión de los genes DCT, MLANA y TYRP1 en la piel que presentó pigmento (normal y repigmentada), en comparación con su contraparte despigmentada (piel con vitiligo y que no repigmentó después de tratamiento). Además, se obtuvo un grupo de genes con utilidad predictiva para respuesta a tratamiento, relacionados con la pigmentación (DCT y TYRP1), y con la respuesta al estrés oxidativo (CCBL2 y GPD1); por otra parte, si se considera el tipo de vitiligo vulgar de acuerdo a su progresión de la enfermedad como factor determinante en la respuesta a tratamiento, el gen MC1R, puede ser de utilidad para diferenciar la respuesta entre grupos activos y estables. Finalmente, el análisis utilizando la aplicación TruSeq Targeted RNA de Illumina sugiere a los genes participantes en la pigmentación de la piel (DCT, TYRP1 y MLANA), regulación del estrés oxidativo (GPD1), apoptosis (CSNK1G3, BAX, BCL3 CFLAR, CASP7 y CASP8) y la ruta de las MAP Cinasas (MAPK1) relacionada con la proliferación y diferenciación celular, como potenciales marcadores moleculares con utilidad pronostica de respuesta a tratamiento. Financiamiento: Este proyecto contó con financiamiento del “Programa de Apoyo a la Investigación Científica y Tecnológica” (PAICyT - UANL) con número CS879-11, y de recursos propios del Servicio de Dermatología, Hospital Universitario Dr. José Eleuterio González, UANL, MTY, Nuevo León México.Vitiligo belongs to a family of skin or acquired congenital diseases, characterized by a lack of pigmentation in the skin as macules or well-defined spots, a chromic or hypo chromic, in which there has been the deterioration of the function of melanocytes, the skin cell responsible for melanin production. Although it has been reported that this disease affects between 0.1 and 2% of the world population, its prevalence varies considerably among the population or ethnic group analyzed. The causes of vitiligo are complex and have not yet been fully understood. Several hypotheses and theories have been developed to explain the process of depigmentation, or even destruction of melanocytes that occurs in this pathology. However, it does not explain the full spectrum of this disease. Studies to date have focused on analyzing the importance of the melanocyte as the producer of pigment and describe how alterations in it have resulted in the development of vitiligo. However most of these have been made by cell culture methods, which can alter the actual cell expression patterns, in comparison to the ones present the tissue of origin, there for the study of the skin’s microenvironment and analysis of the functional status of cellular components can help determine the best therapeutic alternative in repigmentation. Objective: To identify on the skin of vitiligo patients before and after undergoing phototherapy with UVB-nb, changes in patterns of gene expression with prognostic value. Population and Methodology: Comparative, prospective, blinded to assessor, experimental and Longitudinal Study. Forty-five subjects with Vitiligo Vulgaris (21 females and 24 males; 23 with active form of vulgar vitiligo and 22 with its stable form) from the State of Nuevo Leon, Mexico, who attended the Dermatology Department of the University Hospital Jose Eleuterio Gonzalez of the Universidad Autonoma de Nuevo Leon (UANL), Monterrey (MTY), Nuevo Leon, Mexico, after signing the informed consent, underwent treatment UVB-nb. Prior to treatment, blood samples were obtained for biochemical tests at the Central Laboratory and also at the Department of Endocrinology, Hospital Universitario UANL. In addition two skin biopsies were taken at 4 mm punch in each patient, corresponding to skin lesions and healthy skin respectively. It was later used UVB -nb equipment phototherapy for each patient, 2 times a week, 48 total sessions. At the end of the protocol control biopsies were taken, one in repigmented tissue and the other in tissue that did not undergo treatment response. Skin biopsies were used to extract mRNA by RNAeasy MINIKIT of QIAGEN, it was quantified and used for expression analysis using RNA-Seq (expression analysis by massive sequencing of RNA) in genes involved in pathways of pigmentation skin, apoptosis, oxidative stress, signal transduction and cell survival, in order to identify changes associated with the state of the patient's skin with treatment prognostic utility. The molecular analysis and expression profiling of skin biopsies were performed in the Unit of Molecular Biology, Genomics and Sequencing (UBMGyS) of the Center for Research and Development in Health Sciences (CIDICS) UANL, MTY, Mexico. Clinical results of treatment response were evaluated by two independent investigators and blinded to the results of the massive sequencing of RNA by TruSeq of skin samples. Finally the statistical analysis of expression levels and their relationship with clinical parameters, biochemical and response to treatment was performed by R Bioconductor, SPSS and BaseSpace of Illumina. Results: It was observed that most patients develop the disease before age 30 years, and that the development of the disease is related to a family history of the disease or others of immune nature. The correlation between the clinical results, biochemical and of expression allowed to identify that: low levels of TSH, and high weight in patients can lead to a poor response to UVB-nb treatment. Furthermore expression analysis allowed to detect significant differences in the expression patterns of DCT, MLANA and TYRP1 genes in skin that presented pigment (normal and repigmented) compared with depigmented counterparts (skin with vitiligo and none repigmented skin after treatment). Also a group of genes with predictive utility for response to treatment related pigmentation (DCT and TYRP1), and the response to oxidative stress (CCBL2 and GPD1) was obtained; on the other hand if you consider the kind of vulgar vitiligo according to their disease progression as a determining factor in the response to treatment, the MC1R gene, it may be useful to differentiate between active and stable response groups. Finally analysis using TruSeq Targeted RNA Illumina application suggests participants genes in skin pigmentation (DCT TYRP1 and MLANA), regulation of oxidative stress (GPD1), apoptosis (CSNK1G3, BAX, BCl3 CFLAR, CASP7 and CASP8 ) and the path of MAP kinases (MAPK1) related to cell proliferation and differentiation, as potential molecular markers useful to predict response to treatment. Funding: This project was funded by the "Support Program for Scientific and Technological Research" (PAICYT - UANL) CS879-11 number, and resources of the Department of Dermatology, Hospital Universitario Dr. Jose Eleuterio Gonzalez, UANL, MTY , Nuevo Leon, Mexico

Pigmented purpura and cutaneous vascular occlusion syndromes

Purpura is defined as a visible hemorrhage in the skin or mucosa, which is not evanescent upon pressure. Proper classification allows a better patient approach due to its multiple diagnoses. Purpuras can be categorized by size, morphology, and other characteristics. The course varies according to the etiology, as do the diagnostic approach and treatment. This review discusses pigmented purpuras and some cutaneous vascular occlusion syndromes

The synapsin gene family in basal chordates: evolutionary perspectives in metazoans

<p>Abstract</p> <p>Background</p> <p>Synapsins are neuronal phosphoproteins involved in several functions correlated with both neurotransmitter release and synaptogenesis. The comprehension of the basal role of the synapsin family is hampered in vertebrates by the existence of multiple synapsin genes. Therefore, studying homologous genes in basal chordates, devoid of genome duplication, could help to achieve a better understanding of the complex functions of these proteins.</p> <p>Results</p> <p>In this study we report the cloning and characterization of the <it>Ciona intestinalis </it>and amphioxus <it>Branchiostoma floridae </it>synapsin transcripts and the definition of their gene structure using available <it>C. intestinalis </it>and <it>B. floridae </it>genomic sequences. We demonstrate the occurrence, in both model organisms, of a single member of the synapsin gene family. Full-length synapsin genes were identified in the recently sequenced genomes of phylogenetically diverse metazoans. Comparative genome analysis reveals extensive conservation of the SYN locus in several metazoans. Moreover, developmental expression studies underline that synapsin is a neuronal-specific marker in basal chordates and is expressed in several cell types of PNS and in many, if not all, CNS neurons.</p> <p>Conclusion</p> <p>Our study demonstrates that synapsin genes are metazoan genes present in a single copy per genome, except for vertebrates. Moreover, we hypothesize that, during the evolution of synapsin proteins, new domains are added at different stages probably to cope up with the increased complexity in the nervous system organization. Finally, we demonstrate that protochordate synapsin is restricted to the post-mitotic phase of CNS development and thereby is a good marker of postmitotic neurons.</p

Laser micro-polishing of stainless steel for antibacterial surface applications

In this work laser micro polishing (LMP) of cold rolled 0.3 mm thick 304 stainless steel with a pulsed fibre laser is studied, for applications where antibacterial properties are required. Due to its production method, the initial surface roughness of the tested material was considerably low (Sa=85.3±2.8 µm), rendering a demanding case for the laser polishing process. Accordingly, process feasibility under three different atmospheric conditions, namely ambient, Ar and N2 atmosphere, was investigated. A large set of process parameter combinations was tested and initial analysis was carried out to identify the polishing feasibility by inspection under an optical microscope. Once the feasibility window was determined, a primary characterization was made on selected surfaces for roughness and waviness. Results show that in some process conditions belonging to the explored feasibility range, surface roughness could be decreased by 50%

Laparoscopic management of fallopian tube prolapse masquerading as adenocarcinoma of the vagina in a hysterectomized woman

BACKGROUND: Fallopian tube prolapse as a complication of abdominal hysterectomy is a rare occurrence. A case with fallopian tube prolapse was managed by a combined vaginal and laparoscopic approach and description of the operative technique is presented. CASE PRESENTATION: A 39-year-old woman with vaginal prolapse of the fallopian tube after total abdominal hysterectomy presented with an incorrect diagnosis of adenocarcinoma of the vaginal apex. The prolapsed tube and cystic ovary were removed by vaginal and laparoscopic approach. The postoperative course went well. CONCLUSIONS: Early or late fallopian tube prolapse can occur after total abdominal hysterectomy and vaginal hysterectomy. Symptoms consist of persistent blood loss or leukorrhea, dyspareunia and chronic pelvic pain. Vaginal removal of prolapsed tube with laparoscopic surgery may be a suitable treatment. The abdominal or vaginal approach used in surgical correction of prolapsed tubes must be decided in each case according to the patient's individual characteristics

Investigação sobre usos e manuseios de produtos saneantes por alunos, professores e donas de casa da região metropolitana de São Paulo - Brasil

Diariamente a população está rodeada de produtos e materiais químicos, embora pensem que este assunto seja tão distante da vida de cada um. O objetivo da pesquisa foi investigar a percepção dos entrevistados a respeito dos produtos saneantes. A metodologia foi baseada em um estudo de diagnóstico com caráter quantitativo, onde foi utilizado questionário estruturado, para obter dados a respeito da utilização de produtos de limpeza. O universo pesquisado foi distribuído em grupos: alunos e professores da Universidade Federal de São Paulo e Donas de Casa, da região do ABC e Diadema. Os produtos de limpeza contendo cloro ativo são utilizados pela maioria dos respondentes 3 vezes por semana e, 61 % indicaram que o cloro é prejudicial ao meio ambiente. Os respondentes apontaram disponibilidade à mudança de hábito, para que o meio ambiente seja menos prejudicado e assim, preservado

Silk fibroin microgels as a platform for cell microencapsulation

: Cell microencapsulation has been utilized for years as a means of cell shielding from the external environment while facilitating the transport of gases, general metabolites, and secretory bioactive molecules at once. In this light, hydrogels may support the structural integrity and functionality of encapsulated biologics whereas ensuring cell viability and function and releasing potential therapeutic factors once in situ. In this work, we describe a straightforward strategy to fabricate silk fibroin (SF) microgels (µgels) and encapsulate cells into them. SF µgels (size ≈ 200 µm) were obtained through ultrasonication-induced gelation of SF in a water-oil emulsion phase. A thorough physicochemical (SEM analysis, and FT-IR) and mechanical (microindentation tests) characterization of SF µgels were carried out to assess their nanostructure, porosity, and stiffness. SF µgels were used to encapsulate and culture L929 and primary myoblasts. Interestingly, SF µgels showed a selective release of relatively small proteins (e.g., VEGF, molecular weight, MW = 40 kDa) by the encapsulated primary myoblasts, while bigger (macro)molecules (MW = 160 kDa) were hampered to diffusing through the µgels. This article provided the groundwork to expand the use of SF hydrogels into a versatile platform for encapsulating relevant cells able to release paracrine factors potentially regulating tissue and/or organ functions, thus promoting their regeneration