In vivo movement of retinoblastoma-related protein (RBR) towards cytoplasm during mitosis in Arabidopsisthaliana.

Retinoblastoma protein is central in signaling networks of fundamental cell decisions such as proliferation and differentiation in all metazoans and cancer development. Immunostaining and biochemical evidence demonstrated that during interphase retinoblastoma protein is in the nucleus and is hypophosphorylated, and during mitosis is in the cytoplasm and is hyperphosphorylated. The purpose of this study was to visualize in vivo in a non-diseased tissue, the dynamic spatial and temporal nuclear exit toward the cytoplasm of this protein during mitosis and its return to the nucleus to obtain insights into its potential cytosolic functions. Using high-resolution time-lapse images from confocal microscopy, we tracked in vivo the ortholog in plants the RETINOBLASTOMA RELATED (RBR) protein tagged with Green Fluorescent Protein (GFP) in Arabidopsis thaliana's root. RBR protein exits from dense aggregates in the nucleus before chromosomes are in prophase in less than 2 min, spreading outwards as smaller particles projected throughout the cytosol during mitosis like a diffusive yet controlled event until telophase, when the daughter's nuclei form; RBR returns to the nuclei in coordination with decondensing chromosomal DNA forming new aggregates again in punctuated larger structures in each corresponding nuclei. We propose RBR diffused particles in the cytoplasm may function as a cytosolic sensor of incoming signals, thus coordinating re-aggregation with DNA is a mechanism by which any new incoming signals encountered by RBR may lead to a reconfiguration of the nuclear transcriptomic context. The small RBR diffused particles in the cytoplasm may preserve topologic-like properties allowing them to aggregate and restore their nuclear location, they may also be part of transient cytoplasmic storage of the cellular pre-mitotic transcriptional context, that once inside the nuclei may execute both the pre mitosis transcriptional context as well as new transcriptional instructions.

Curative activity of postharvest GRAS salt treatments to control citrus sour rot caused by Geotrichum citri-aurantii.

The effectiveness of the "generally recognized as safe" (GRAS) salts potassium sorbate (PS), sodium benzoate (SB), sodium ethylparaben (SEP) and sodium methylparaben (SMP) to control sour rot, caused by Geotrichum citri-aurantii, was assessed by dipping economically important citrus species and cultivars in aqueous solutions for 30, 60 or 150 s at 20 °C, followed by examination after 8 d of storage at 28 °C. Curative activity was determined because the fruit were inoculated 24 h prior to treatment. Dipping fruit for 60 s in SMP (200 mM), SEP (200 mM) or SB (3% w/v) were very effective and reduced sour rot incidence and severity by up to 90%. Their effectiveness was similar or superior to that of the conventional fungicide propiconazole (PCZ). In contrast, PS (200 mM) did not control sour rot on 'Oronules' or 'Ortanique' mandarins, but it reduced sour rot incidence on 'Barnfield' oranges by 50% compared to inoculated, water-treated control fruit. Sour rot was better controlled on oranges than on mandarins. Furthermore, heating the solutions to 50 °C enhanced their effectiveness, while post-treatment rinsing of the fruit with tap water reduced their effectiveness. Dipping 'Valencia Late' oranges in SB (3% w/v) or SMP (200 mM) for 60 s followed by long storage for up to 8 weeks at 5 °C and 90% RH, reduced sour rot incidence from 55% among water-treated control fruit to 2 to 6%, and matched the effectiveness of PCZ. No fruit in any test were visibly harmed. Both SB and SMP salts could be potential alternatives to conventional fungicides, such as PCZ or guazatine, for the integrated postharvest management of citrus sour rot.

MicroRNA Profiling in Wilms Tumor: Identification of Potential Biomarkers.

Wilms tumor (WT) is the most frequently diagnosed malignant renal tumor in children. With current treatments, ~90% of children diagnosed with WT survive and generally present with tumors characterized by favorable histology (FHWT), whereas prognosis is poor for the remaining 10% of cases where the tumors are characterized by cellular diffuse anaplasia (DAWT). Relatively few studies have investigated microRNA-related epigenetic regulation and its relationship with altered gene expression in WT. Here, we aim to identify microRNAs differentially expressed in WT and describe their expression in terms of cellular anaplasia, metastasis, and association with the main genetic alterations in WT to identify potential prognostic biomarkers. Expression profiling using TaqMan low-density array was performed in a discovery cohort consisting of four DAWT and eight FHWT samples. Relative quantification resulted in the identification of 109 (48.7%) microRNAs differentially expressed in both WT types. Of these, miR-10a-5p, miR-29a-3p, miR-181a-5p, miR-200b-3p, and miR-218-5p were selected and tested by RT-qPCR on a validation cohort of 53 patient samples. MiR-29a and miR-218 showed significant differences in FHWT with low (P = 0.0018) and high (P = 0.0131) expression, respectively. To discriminate between miRNA expression FHWTs and healthy controls, the receiver operating characteristic (ROC) curves were obtained; miR-29a AUC was 0.7843. Furthermore, low expression levels of miR-29a and miR-200b (P = 0.0027 and P = 0.0248) were observed in metastatic tumors. ROC curves for miR-29a discriminated metastatic patients (AUC = 0.8529) and miR-200b (AUC = 0.7757). To confirm the differences between cases with poor prognosis, we performed in situ hybridization for three microRNAs in five DAWT and 17 FHWT samples, and only significant differences between adjacent tissues and FHWT tumors were found for miR-181a, miR-200b, and miR-218, in both total pixels and nuclear analyses. Analysis of copy number variation in genes showed that the most prevalent alterations were WTX (47%), IGF2 (21%), 1q (36%) gain, 1p36 (16%), and WTX deletion/1q duplicate (26%). The five microRNAs evaluated are involved in the Hippo signaling pathway and participate in Wilms tumor development through their effects on differentiation, proliferation, angiogenesis, and metastasis.

Discovery of a transcriptomic core of genes shared in 8 primary retinoblastoma with a novel detection score analysis.

PURPOSE: Expression microarrays are powerful technology that allows large-scale analysis of RNA profiles in a tissue; these platforms include underexploited detection scores outputs. We developed an algorithm using the detection score, to generate a detection profile of shared elements in retinoblastoma as well as to determine its transcriptomic size and structure. METHODS: We analyzed eight briefly cultured primary retinoblastomas with the Human transcriptome array 2.0 (HTA2.0). Transcripts and genes detection scores were determined using the Detection Above Background algorithm (DABG). We used unsupervised and supervised computational tools to analyze detected and undetected elements; WebGestalt was used to explore functions encoded by genes in relevant clusters and performed experimental validation. RESULTS: We found a core cluster with 7,513 genes detected and shared by all samples, 4,321 genes in a cluster that was commonly absent, and 7,681 genes variably detected across the samples accounting for tumor heterogeneity. Relevant pathways identified in the core cluster relate to cell cycle, RNA transport, and DNA replication. We performed a kinome analysis of the core cluster and found 4 potential therapeutic kinase targets. Through analysis of the variably detected genes, we discovered 123 differentially expressed transcripts between bilateral and unilateral cases. CONCLUSIONS: This novel analytical approach allowed determining the retinoblastoma transcriptomic size, a shared active transcriptomic core among the samples, potential therapeutic target kinases shared by all samples, transcripts related to inter tumor heterogeneity, and to determine transcriptomic profiles without the need of control tissues. This approach is useful to analyze other cancer or tissue types.

Circulating miRNome detection analysis reveals 537 miRNAS in plasma, 625 in extracellular vesicles and a discriminant plasma signature of 19 miRNAs in children with retinoblastoma from which 14 are also detected in corresponding primary tumors.

miRNAs regulate post-transcriptional gene expression in metazoans, and thus are involved in many fundamental cellular biological processes. Extracellular miRNAs are also found in most human biofluids including plasma. These circulating miRNAs constitute a long distance inter cellular communication system and are potentially useful biomarkers. High throughput technologies like microarrays are able to scan a complete miRNome providing useful detection scores that are underexplored. We proposed to answer how many and which miRNAs are detectable in plasma or extracellular vesicles as these questions have not yet been answered. We set out to address this knowledge gap by analyzing the mirRNome in plasma and corresponding extracellular vesicles (EVs) from 12 children affected by retinoblastoma (Rb) a childhood intraocular malignant tumor, as well as from 12 healthy similarly aged controls. We calculated an average of 537 detectable miRNAs in plasma and 625 in EVs. The most miRNA enriched compartment were EVs from Rb cases with an average of 656 detectable elements. Using hierarchical clustering with the detection scores, we generated broad detection mirnome maps and identified a plasma signature of 19 miRNAs present in all Rb cases that is able to discriminate cases from controls. An additional 9 miRNAs were detected in all the samples; within this group, miRNA-5787 and miRNA-6732-5p were highly abundant and displayed very low variance across all the samples, suggesting both are good candidates to serve as plasma references or normalizers. Further exploration considering participant's sex, allowed discovering 5 miRNAs which corresponded only to females and 4 miRNAs corresponding only to males. Target and pathway analysis of these miRNAs revealed hormonal function including estrogen, thyroid signaling pathways and testosterone biosynthesis. This approach allows a comprehensive unbiased survey of a circulating miRNome landscape, creating the possibility to define normality in mirnomic profiles, and to locate where in these miRNome profiles promising and potentially useful circulating miRNA signatures can be found.

Live Attenuated Salmonella enterica Expressing and Releasing Cell-Permeable Bax BH3 Peptide Through the MisL Autotransporter System Elicits Antitumor Activity in a Murine Xenograft Model of Human B Non-hodgkin's Lymphoma.

The survival of patients with non-Hodgkin's lymphoma (NHL) has substantially improved with current treatments. Nevertheless, the appearance of drug-resistant cancer cells leads to patient relapse. It is therefore necessary to find new antitumor therapies that can completely eradicate transformed cells. Chemotherapy-resistant cancer cells are characterized by the overexpression of members of the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein family, such as Bcl-XL, Bcl-2, and Mcl-1. We have recently shown that peptides derived from the BH3 domain of the pro-apoptotic Bax protein may antagonize the anti-apoptotic activity of the Bcl-2 family proteins, restore apoptosis, and induce chemosensitization of tumor cells. In this study, we investigated the feasibility of releasing this peptide into the tumor microenvironment using live attenuated Salmonella enterica, which has proven to be an ally in cancer therapy due to its high affinity for tumor tissue, its ability to activate the innate and adaptive antitumor immune responses, and its potential use as a delivery system of heterologous molecules. Thus, we expressed and released the cell-permeable Bax BH3 peptide from the surface of Salmonella enterica serovar Typhimurium SL3261 through the MisL autotransporter system. We demonstrated that this recombinant bacterium significantly decreased the viability and increased the apoptosis of Ramos cells, a human B NHL cell line. Indeed, the intravenous administration of this recombinant Salmonella enterica elicited antitumor activity and extended survival in a xenograft NHL murine model. This antitumor activity was mediated by apoptosis and an inflammatory response. Our approach may represent an eventual alternative to treat relapsing or refractory NHL.

Expression of YY1 in Wilms tumors with favorable histology is a risk factor for adverse outcomes.

AIM: To investigate the role of the transcription factor YY1 in Wilms tumor (WT). PATIENTS & METHODS: We measured YY1 expression using tissue microarray from patients with pediatric renal tumors, mainly WT and evaluated correlations with the predicted clinical evolution. YY1 expression was measured using immunohistochemical and protein expression was determined by digital pathology. RESULTS & CONCLUSION: YY1 significantly increased in WT patients. In addition, an increase in YY1 expression had a greater risk of adverse outcomes in WT patients with favorable histology. YY1 expression was higher in the blastemal component of tumors, and high nuclear expression positively correlated with metastasis. YY1 may be considered as a metastasis risk factor in WT.

miRNome landscape analysis reveals a 30 miRNA core in retinoblastoma.

BACKGROUND: miRNAs exert their effect through a negative regulatory mechanism silencing expression upon hybridizing to their target mRNA, and have a prominent position in the control of many cellular processes including carcinogenesis. Previous miRNA studies on retinoblastoma (Rb) have been limited to specific miRNAs reported in other tumors or to medium density arrays. Here we report expression analysis of the whole miRNome on 12 retinoblastoma tumor samples using a high throughput microarray platform including 2578 mature miRNAs. METHODS: Twelve retinoblastoma tumor samples were analyzed using an Affymetrix platform including 2578 mature miRNAs. We applied RMA analysis to normalize raw data, obtained categorical data from detection call values, and also used signal intensity derived expression data. We used Diana-Tools-microT-CDS to find miRNA targets and ChromDraw to map miRNAs in chromosomes. RESULTS: We discovered a core-cluster of 30 miRNAs that were highly expressed in all the cases and a cluster of 993 miRNAs that were uniformly absent in all cases. Another 1022 miRNA were variably present in the samples reflecting heterogeneity between tumors. We explored mRNA targets, pathways and biological processes affected by some of these miRNAs. We propose that the core-cluster of 30 miRs represent miRNA machinery common to all Rb, and affecting most pathways considered hallmarks of cancer. In this core, we identified miR-3613 as a potential and critical down regulatory hub, because it is highly expressed in all the samples and its potential mRNA targets include at least 36 tumor suppressor genes, including RB1. In the variably expressed miRNA, 36 were differentially expressed between males and females. Some of the potential pathways targeted by these 36 miRNAs were associated with hormonal production. CONCLUSION: These findings indicate that Rb tumor samples share a common miRNA expression profile regardless of tumor heterogeneity, and shed light on potential novel therapeutic targets such as mir-3613 This is the first work to delineate the miRNA landscape in retinoblastoma tumor samples using an unbiased approach.

Distribution of Native Lactic Acid Bacteria in Wineries of Queretaro, Mexico and Their Resistance to Wine-Like Conditions.

Native lactic acid bacteria (LAB) are capable of growing during winemaking, thereby strongly affecting wine quality. The species of LAB present in musts, wines during malolactic fermentation (MLF), and barrels/filters were investigated in wineries from the emerging wine region of Queretaro, México using multiplex PCR and culture. The resistance to wine-like conditions (WLC): ethanol (10, 12, and 13%), SO2 (30 mg⋅l-1), and low pH (3.5) of native LAB strains was also studied. Five species were detected within 61 samples obtained: Oenococcus oeni, Lactobacillus plantarum, Pediococcus parvulus, Lactobacillus hilgardi, and Lactobacillus brevis. Four species (excepting L. brevis) were found in must; O. oeni and P. parvulus were ubiquitous in wine and L. plantarum and L. brevis were mainly present at the initial stage of MLF, while L. hilgardii was mostly detected at the advanced stage. Furthermore, some species detected in barrel/filter, prove them to be hazardous reservoirs. From 822 LAB isolates, only 119 resisted WLC with 10% ethanol; the number of strains able to grow in WLC with 13% ethanol decreased approximately by 50%, O. oeni being the most versatile species with 65% of resistant isolates, while Lactobacillus spp. and P. parvulus were the most strongly affected, especially those recovered from barrel/filter, with less than 10% of resistant isolates. This study evidences the presence of local strains able to be used as starter cultures, and also enabled the assessment of the risks derived from the presence of spoilage LAB strains resistant to WLC.

O treinamento de pais e cuidadores: ensinando a educar e promovendo a saúde mental / Parents and caregivers training: teaching to educate and promoting mental health

Educar sem usar de violência pode ser tarefa difícil, principalmente para quem teve um modelo de educação violenta e tem que enfrentar problemas de comportamento de seus filhos como desobediência, mentira e agressividade. Para evitar a repetição de padrões educativos violentos, existem técnicas de educação pacíficas que podem ser ensinadas, como o treinamento de pais (TP). Este artigo revisou a literatura sobre as experiências brasileiras de treinamento de pais, enfocando as técnicas utilizadas e os seus resultados, verificando a sua aplicabilidade em programas públicos de saúde mental. Foram realizados levantamentos bibliográficos nas bases de dados SCIELO e PUBMED/MEDLINE utilizando-se as palavras-chaves "treinamento pais"; "violência"; "saúde mental"; "TEPT". No total, foram encontrados cinco estudos abordando o treinamento de pais, sendo que apenas um deles utilizou TP em ambulatório de Psiquiatria Infantil. Foi também descrita neste artigo a experiência clínica do ambulatório de trauma da UFMG. Concluiu-se que houve coerência entre os resultados de artigos que relataram o uso de TP e entre estes resultados e a experiência clínica do ambulatório de trauma e que o treinamento de pais e cuidadores pode favorecer que o contexto familiar seja mediador da saúde mental de crianças e adolescentes, ao minimizar as práticas educativas violentas no ambiente doméstico.

Educate without using violence may be a hard task, mainly to those who have had a violent model of education and have to face the problems of their own children behavior, such as disobedience, lies and aggressiveness. To avoid repeating violent educational patterns, there are techniques of peaceful education that can be taught, such as parental training (PT). This article reviewed the literature about Brazilian experiences on parents training, focusing the used techniques and their results, checking their applicability on mental health public programs. Bibliographic surveys on the database SCIELO and PUBMED/MEDLINEwere carried out using the key words "parents training"; "violence"; "mental health"; "TEPT". In total, there were five studies addressing parents training, and only one of them used PT in outpatient Child Psychiatry. This article also describes the clinical experience of UFMG trauma clinic. The conclusion was that there was consistency between the results of articles that reported the use of PT, and between these results and the clinical experience of outpatients trauma clinic, and that parents and caregivers training can support the family background as a mediator of mental health of children and teenagers, while minimizing the violence in the educational parenting practices in the home environment.

Um serviço de saúde especializado em transtorno de estresse pós-traumático: um modelo cognitivo-comportamental / A specialized health care service in post-traumatic stress syndrome: a cognitive-behavioral model

A violência, um problema social de saúde, causa seqüelas na saúde mental e física das populações e gera demanda expressiva aos serviços de saúde. A violência pode ser um evento traumático, uma situação social crítica que compromete a saúde mental e o desenvolvimento integral dos sujeitos, manifestando-se clinicamente como transtorno de estresse pós-traumático (TEPT). O objetivo com este estudo é discutir o arcabouço científico de um serviço de saúde mental ambulatorial em hospital geral adequado metodologicamente para a terapia de vítimas de violência com TEPT freqüentemente acompanhado de declínio no desempenho cognitivo, sensações persistente de culpa e/ou hostilidade, raiva, além de comportamento de oposição e problemas de memória e atenção. A violência, como informação oriunda do meio social, deve ser interpretada cognitiva e emocionalmente para ser operativa. Neste artigo conclui-se que falhas no processamento de informação social podem originar o TEPT. Nessa perspectiva, foi criado o Ambulatório do Trauma, em parceria com o curso de psicologia da FUMEC e o serviço de psiquiatria do Hospital das Clínicas da UFMG a fim de atender, em um referencial sociocognitivo-comportamental, a população vítima de TEPT em Belo Horizonte(AU)

Comparación de la biopsia por aspiración con aguja fina y la biopsia quirúrgica en el diagnóstico de tumores en niños / Comparison between diagnosisi by fine needle aspiration biopsy and conventional biopsy in children's tumors

Introducción. La biopsia por aspiración con aguja fina (BAAF) es un método de diagnóstico útil, inocuo y rápido para el estudio inicial de los tumores superficiales y profundos que ha sido utilizado cada vez con mayor frecuencia en niños. Fue propósito de este trabajo evaluar la precisión diagnóstica del procedimiento en el diagnóstico inicial de tumores en niños. Material y métodos. Se revisaron todas las BAAF diagnosticadas en un período de 24 meses y se seleccionaron aquellas con diagnóstico de tumor o masa tumoral que tuvieran además estudio histopatológico subsecuente ya sea mediante biopsia o extirpación total de la lesión. Se compararon los diagnósticos de ambos procedimientos y se juzgó conveniente calcular la sensibilidad y especificidad. Resultados. De 89 BAAF registradas en el período de estudio solamente 34 tuvieron estudio histopatológico subsecuente. La gran mayoría de las lesiones correspondieron a neoplasias malignas. En 30 casos el diagnóstico de la BAAF estuvo de aucerdo con el de la biopsia quirúrgica y no hubo acuerdo en 4. La sensibilidad del método fue del 97 por ciento y la especificidad del 33 por ciento. Conclusiones. Los resultados de este estudio indican que la BAAF es confiable para el diagnóstico inicial de tumores superficiales y profundos en niños

Sesión clinicopatológica: diarreas por shigella, desnutrición y choque hipovolémico / Clinicopathological session: diarrheas by shiguella, malnutrition and wypovolemic shock

Se describe un caso de diarrea crónica causada por shigella en un paciente de siete meses de edad, masculino, procedente de Ciudad Altamirano, Guerrero, México, madre de 24 años de edad, sana, padre de 2 años, campesino, escolaridad sexto año de primaria, alcoholismo positivo y, tres hermanos sanos de seis, cuatro y dos años. Ingresó el 8 de febrero de 1988 y fue dado de alta el 14 del mismo mes