Selective cytotoxic and anti-metastatic activity in DU-145 prostate cancer cells induced by Annona muricata L. bark extract and phytochemical, annonacin.

BACKGROUND: Annona muricata L. was identified as a popular medicinal plant in treatment regimens among cancer patients in Jamaica by a previously conducted structured questionnaire. Ethnomedically used plant parts, were examined in this study against human prostate cancer cells for the first time and mechanisms of action elucidated for the most potent of them, along with the active phytochemical, annonacin. METHODS: Nine extracts of varying polarity from the leaves and bark of A. muricata were assessed initially for cytotoxicity using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on PC-3 prostate cancer cells and the ethyl acetate bark (EAB) extract was identified as the most potent. EAB extract was then standardized for annonacin content using High-performance Liquid Chromatography - Mass Spectrometry (HPLC-MS) and shown to be effective against a second prostate cancer cell line (DU-145) also. The mode of cell death in DU-145 cells were assessed via several apoptotic assays including induction of increased reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential, activation of caspases and annexin V externalization combined with morphological observations using confocal microscopy. In addition, the potential to prevent metastasis was examined via inhibition of cell migration, vascular endothelial growth factor (VEGF) and angiogenesis using the chorioallantoic membrane assay (CAM). RESULTS: Annonacin and EAB extract displayed selective and potent cytotoxicity against the DU-145 prostate carcinoma cells with IC50 values of 0.1 ± 0.07 µM and 55.501 ± 0.55 µg/mL respectively, without impacting RWPE-1 normal prostate cells, in stark contrast to chemotherapeutic docetaxel which lacked such selectivity. Docetaxel's impact on the cancerous DU-145 was improved by 50% when used in combination with EAB extract. Insignificant levels of intracellular ROS content, depolarization of mitochondrial membrane, Caspase 3/7 activation, annexin V content, along with stained morphological evaluations, pointed to a non-apoptotic mode of cell death. The extract at 50 µg/mL deterred cell migration in the wound-healing assay, while inhibition of angiogenesis was displayed in the CAM and VEGF inhibition assays for both EAB (100 µg /mL) and annonacin (0.5 µM). CONCLUSIONS: Taken together, the standardized EAB extract and annonacin appear to induce selective and potent cell death via a necrotic pathway in DU-145 cells, while also preventing cell migration and angiogenesis, which warrant further examinations for mechanistic insights and validity in-vivo.

Using Telemedicine Technology to Assess Physician Outpatient Teaching.

BACKGROUND AND OBJECTIVES: Video conferencing technology (telemedicine) can be applied to many settings within the medical community; we assessed the feasibility of its use in conducting observations of faculty at remote family medicine teaching sites. METHODS: We deployed seven telemedicine units to five family medicine residency sites and two observation stations within our division. Practice managers and physician faculty members received on-site training on the basic functionality of the technology, as well as "best practices" and minor troubleshooting techniques. Quick reference guides and other support documents were developed and provided for each site. During the remote faculty observation, two observers simultaneously viewed the resident being precepted, assessing the faculty member using a standardized tool. After the experience, all participants were asked to complete a survey on the usability of the technology. RESULTS: Nineteen observations were successfully conducted from November 2011 to December 2012. From a qualitative perspective, faculty accepted this as a viable means of faculty development. Minor technical hurdles were captured in the survey and improved upon as staff and faculty became more comfortable with the technology and as our technical capabilities allowed. Overall, the technology was rapidly accepted into the practices. CONCLUSIONS: Video teleconferencing represents a valuable tool that contributes to the development of faculty by making observation available to numerous sites, including remote areas that may have been previously challenging to reach due to logistics. Recent improvements in technology should make the process easier and allow more aspects of the encounters to be readily observed.

Substance use and sexual risk among at-risk adolescents in Juiz de Fora, Minas Gerais State, Brazil.

We examined the difference in prevalence of substance use and sexual risk behaviors among at-risk youth participants in programs offered by community-based organizations in Juiz de Fora, Minas Gerais State, Brazil, by gender and organization type (governmental vs. non-governmental). 388 adolescents were recruited from 25 intervention-based organizations servicing at-risk youth between the ages of 12 and 17 in Juiz de Fora. Participants completed a 15-item survey assessing substance use and sexual risk behaviors, along with socio-demographic predictors of these behaviors. Males were more likely to report drug and alcohol use associated with homelessness and abandonment. Females were more likely to report sexual risk taking associated with neighborhood crime. Participants from non-governmental organizations were less likely to engage in all measured risk behaviors. The present analysis points to the need to understand how Brazil's Child and Adolescent Act is being applied and the implications this has for intervention and the promotion of positive health outcomes for young people.

Substance use and sexual risk among at-risk adolescents in Juiz de Fora, Minas Gerais State, Brazil / O uso de substâncias e o risco sexual entre adolescentes em situação de risco social em Juiz de Fora, Minas Gerais, Brasil / Consumo de sustancias estupefacientes y riesgo sexual entre adolescentes en situaciones de riesgo en Juiz de Fora, Minas Gerais, Brasil

We examined the difference in prevalence of substance use and sexual risk behaviors among at-risk youth participants in programs offered by community-based organizations in Juiz de Fora, Minas Gerais State, Brazil, by gender and organization type (governmental vs. non-governmental). 388 adolescents were recruited from 25 intervention-based organizations servicing at-risk youth between the ages of 12 and 17 in Juiz de Fora. Participants completed a 15-item survey assessing substance use and sexual risk behaviors, along with socio-demographic predictors of these behaviors. Males were more likely to report drug and alcohol use associated with homelessness and abandonment. Females were more likely to report sexual risk taking associated with neighborhood crime. Participants from non-governmental organizations were less likely to engage in all measured risk behaviors. The present analysis points to the need to understand how Brazil’s Child and Adolescent Act is being applied and the implications this has for intervention and the promotion of positive health outcomes for young people.

Examinou-se a diferença na prevalência do uso de drogas e comportamento sexual de risco entre os jovens de alto risco em programas oferecidos por organizações localizadas na comunidade de Juiz de Fora, Minas Gerais, Brasil .Foram recrutados 388 adolescentes de 25 organizações de intervenções preventivas que servem jovens de alto risco entre as idades de 12-17. Participantes completaram um questionário com 15 perguntas que avalia o uso de substâncias e o comportamento sexual de alto risco, juntamente com indicadores sociodemográficos. Os adolescentes do sexo masculino foram mais propensos a relacionar o uso de drogas e álcool com a falta de moradia e abandono. As adolescentes do sexo feminino foram mais propensas a reportar comportamentos sexuais de alto risco em associação com o crime comum. Participantes de ONGs foram menos propensos a se envolver nos comportamentos de alto risco pesquisados. A presente análise aponta para a necessidade de se entender as implicações que isso tem para a intervenção preventiva e a promoção de resultados positivos na saúde dos jovens.

Examinamos las diferencias en la prevalencia de uso de sustancias estupefacientes y el riesgo sexual entre los jóvenes participantes de los programas ofrecidos por las organizaciones de base comunitaria en Juiz de Fora, Minas Gerais, Brasil. Se incorporaron 388 adolescentes de 25 organizaciones de apoyo e intervención a la juventud, que atienden a chicos de entre de 12-17 años en Juiz de Fora. Los participantes completaron un estudio con 15-ítems para evaluar el consumo de sustancias estupefacientes y el riesgo sexual, junto a aspectos socio-demográficos predictores de estos comportamientos. Los varones tenían más probabilidades de uso de drogas y alcohol, asociado al desamparo y abandono. Las mujeres tenían más probabilidades de riesgos sexuales, relacionados con la delincuencia del barrio. Los participantes de las organizaciones no gubernamentales tenían menos probabilidades de comportamientos de riesgo. El presente análisis apunta la necesidad de entender cómo se está aplicando la ley del niño y del adolescente y las consecuencias que tiene para la intervención y la promoción de los resultados positivos para la salud.

Pediatric brainstem gangliogliomas show BRAF(V600E) mutation in a high percentage of cases.

Brainstem gangliogliomas (GGs), often cannot be resected, have a much poorer prognosis than those located in more common supratentorial sites and may benefit from novel therapeutic approaches. Therapeutically targetable BRAF c.1799T>A (p.V600E) (BRAF(V600E) ) mutations are harbored in roughly 50% of collective GGs taken from all anatomical sites. Large numbers of pediatric brainstem GGs, however, have not been specifically assessed and anatomic-and age-restricted assessment of genetic and biological factors are becoming increasingly important. Pediatric brainstem GGs (n = 13), non-brainstem GGs (n = 11) and brainstem pilocytic astrocytomas (PAs) (n = 8) were screened by standard Sanger DNA sequencing of BRAF exon 15. Five of 13 (38%) pediatric GG harbored a definitive BRAF(V600E) mutation, with two others exhibiting an equivocal result by this method. BRAF(V600E) was also seen in five of 11 (45%) non-brainstem GGs and one of eight (13%) brainstem PAs. VE1 immunostaining for BRAF(V600E) showed concordance with sequencing in nine of nine brainstem GGs including the two cases equivocal by Sanger. The equivocal brainstem GGs were subsequently shown to harbor BRAF(V600E) using a novel, more sensitive, RNA-sequencing approach, yielding a final BRAF(V600E) mutation frequency of 54% (seven of 13) in brainstem GGs. BRAF(V600E) -targeted therapeutics should be a consideration for the high percentage of pediatric brainstem GGs refractory to conventional therapies.