Spheroid Model of Mammary Tumor Cells: Epithelial-Mesenchymal Transition and Doxorubicin Response.

Breast cancer is the most prevalent cancer among women worldwide. Therapeutic strategies to control tumors and metastasis are still challenging. Three-dimensional (3D) spheroid-type systems more accurately replicate the features of tumors in vivo, working as a better platform for performing therapeutic response analysis. This work aimed to characterize the epithelial-mesenchymal transition and doxorubicin (dox) response in a mammary tumor spheroid (MTS) model. We evaluated the doxorubicin treatment effect on MCF-7 spheroid diameter, cell viability, death, migration and proteins involved in the epithelial-mesenchymal transition (EMT) process. Spheroids were also produced from tumors formed from 4T1 and 67NR cell lines. MTSs mimicked avascular tumor characteristics, exhibited adherens junction proteins and independently produced their own extracellular matrix. Our spheroid model supports the 3D culturing of cells isolated from mice mammary tumors. Through the migration assay, we verified a reduction in E-cadherin expression and an increase in vimentin expression as the cells became more distant from spheroids. Dox promoted cytotoxicity in MTSs and inhibited cell migration and the EMT process. These results suggest, for the first time, that this model reproduces aspects of the EMT process and describes the potential of dox in inhibiting the metastatic process, which can be further explored.

A Strategy Utilizing Protein-Protein Interaction Hubs for the Treatment of Cancer Diseases.

We describe a strategy for the development of a rational approach of neoplastic disease therapy based on the demonstration that scale-free networks are susceptible to specific attacks directed against its connective hubs. This strategy involves the (i) selection of up-regulated hubs of connectivity in the tumors interactome, (ii) drug repurposing of these hubs, (iii) RNA silencing of non-druggable hubs, (iv) in vitro hub validation, (v) tumor-on-a-chip, (vi) in vivo validation, and (vii) clinical trial. Hubs are protein targets that are assessed as targets for rational therapy of cancer in the context of personalized oncology. We confirmed the existence of a negative correlation between malignant cell aggressivity and the target number needed for specific drugs or RNA interference (RNAi) to maximize the benefit to the patient's overall survival. Interestingly, we found that some additional proteins not generally targeted by drug treatments might justify the addition of inhibitors designed against them in order to improve therapeutic outcomes. However, many proteins are not druggable, or the available pharmacopeia for these targets is limited, which justifies a therapy based on encapsulated RNAi.

miR-138-5p induces aggressive traits by targeting Trp53 expression in murine melanoma cells, and correlates with poor prognosis of melanoma patients.

Deregulation of miRNAs contributes to the development of distinct cancer types, including melanoma, an aggressive form of skin cancer characterized by high metastatic potential and poor prognosis. The expression of a set of 580 miRNAs was investigated in a model of murine melanoma progression, comprising non-metastatic (4C11-) and metastatic melanoma (4C11+) cells. A significant increase in miR-138-5p expression was found in the metastatic 4C11+ melanoma cells compared to 4C11-, which prompted us to investigate its role in melanoma aggressiveness. Functional assays, including anoikis resistance, colony formation, collective migration, serum-deprived growth capacity, as well as in vivo tumor growth and experimental metastasis were performed in 4C11- cells stably overexpressing miR-138-5p. miR-138-5p induced an aggressive phenotype in mouse melanoma cell lines leading to increased proliferation, migration and cell viability under stress conditions. Moreover, by overexpressing miR-138-5p, low-growing and non-metastatic 4C11- cells became highly proliferative and metastatic in vivo, similar to the metastatic 4C11+ cells. Luciferase reporter analysis identified the tumor suppressor Trp53 as a direct target of miR-138-5p. Using data sets from independent melanoma cohorts, miR-138-5p and P53 expression were also found deregulated in human melanoma samples, with their levels negatively and positively correlated with prognosis, respectively. Our data shows that the overexpression of miR-138-5p contributes to melanoma metastasis through the direct suppression of Trp53.

Caracterização dos episódios de queda em uma unidade de cardiologia: estudo retrospectivo / Caracterización de episodios de caídas en una unidad de cardiogolía: estudio restrospectivo / Characterization of falls in a cardiology unit: a retrospective study

Objetivo: Caracterizar os episódios de queda em pacientes internados em uma unidade cardiológica, quanto à ocorrência, fatores relacionados e risco. Método: Estudo descritivo, retrospectivo, por meio da análise de prontuários de pacientes internados em uma unidade de cardiologia que apresentaram episódio de queda entre janeiro de 2015 a dezembro 2016. O risco médio para quedas foi avaliado, conforme a Escala de Morse. Resultados: No período de estudo houve 32 episódios de quedas. A queda foi mais frequente em idosos (81,3%) e naqueles que faziam uso crônico de medicamentos , para controle e tratamento de comorbidades preexistentes. Dentre os fatores de risco, 34,4% apresentavam delirium, comprometimento neurológico e déficit de locomoção. O risco médio para quedas foi classificado como elevado (> 45), 25% das quedas resultaram em algum tipo de dano (leve ou moderado) e ocorreram em períodos matutinos. Conclusão: As contribuições fornecidas pelos registros de eventos adversos, deste estudo, facilitaram a identificação dos fatores de risco, demonstrando a necessidade de se propor intervenções de enfermagem preventivas, uma vez que assumir o evento e identificar suas causas são maneiras de praticar uma assistência de enfermagem segura ao paciente(AU)

Objective: To characterize the episodes of falls in patients hospitalized in a cardiology unit, regarding occurrence, related factors and risk. Method: This is a descriptive, retrospective study, analyzing medical records of patients admitted to a cardiology unit who experienced a fall episode between January 2015 and December 2016. The average risk for falls was assessed according to the Morse Scale. Results: During the study period, there were 32 episodes of falls. Falls were more frequent in the elderly people (81.3%) and in those people who used chronic drugs to control and treat pre-existing comorbidities. Among the risk factors, 34.4% had delirium, neurological impairment and impaired mobility. The average risk for falls was classified as high (> 45), 25% of the falls resulted in some type of damage (mild or moderate) and occurred in the morning. Conclusion: The contributions provided by the records of adverse events in this study facilitated the identification of risk factors, demonstrating the need to propose preventive nursing interventions, since assuming the event and identifying its causes are ways to practice safe nursing care to the patient(AU)

Objetivo: Caracterizar los episodios de caídas en pacientes hospitalizados en una unidad de cardiología, en cuanto a ocurrencias, factores relacionados y riesgo. Método: Estudio descriptivo, retrospectivo, mediante el análisis de las historias clínicas de los pacientes ingresados en una unidad de cardiología que experimentaron un episodio de caída entre enero de 2015 y diciembre de 2016. El riesgo promedio de caídas se evaluó según la escala de Morse. Resultados: Durante el período de estudio hubo 32 episodios de caídas. La caída fue más frecuente en los ancianos (81,3%) y en los que usaban fármacos crónicos para controlar y tratar las comorbilidades preexistentes. Entre los factores de riesgo, el 34,4% presentaba delirio, deterioro neurológico y movilidad reducida. El riesgo promedio de caídas se clasificó como alto (> 45), el 25% de las caídas resultaron en algún tipo de daño (leve o moderado) y ocurrieron en la mañana. Conclusión: Los aportes que brindan los registros de eventos adversos en este estudio facilitaron la identificación de factores de riesgo, demostrando la necesidad de proponer intervenciones de enfermería preventivas, y a que asumir el evento e identificar sus causas son formas de practicar al paciente cuidados de enfermería seguros(AU)

Gene expression and promoter methylation of angiogenic and lymphangiogenic factors as prognostic markers in melanoma.

The high mortality rate of melanoma is broadly associated with its metastatic potential. Tumor cell dissemination is strictly dependent on vascularization; therefore, angiogenesis and lymphangiogenesis play an essential role in metastasis. Hence, a better understanding of the players of tumor vascularization and establishing them as new molecular biomarkers might help to overcome the poor prognosis of melanoma patients. Here, we further characterized a linear murine model of melanoma progression and showed that the aggressiveness of melanoma cells is closely associated with high expression of angiogenic factors, such as Vegfc, Angpt2, and Six1, and that blockade of the vascular endothelial growth factor pathway by the inhibitor axitinib abrogates their tumorigenic potential in vitro and in the in vivo chicken chorioallantoic membrane assay. Furthermore, analysis of The Cancer Genome Atlas data revealed that the expression of the angiogenic factor ANGPT2 (P-value = 0.044) and the lymphangiogenic receptor VEGFR-3 (P-value = 0.002) were independent prognostic factors of overall survival in melanoma patients. Enhanced reduced representation bisulfite sequencing-based methylome profiling revealed for the first time a link between abnormal VEGFC, ANGPT2, and SIX1 gene expression and promoter hypomethylation in melanoma cells. In patients, VEGFC (P-value = 0.031), ANGPT2 (P-value < 0.001), and SIX1 (P-value = 0.009) promoter hypomethylation were independent prognostic factors of shorter overall survival. Hence, our data suggest that these angio- and lymphangiogenesis factors are potential biomarkers of melanoma prognosis. Moreover, these findings strongly support the applicability of our melanoma progression model to unravel new biomarkers for this aggressive human disease.

Intestinal parasitism and related risk factors for primary school students in João Pessoa, northeast Brazil / Parasitismo intestinal e fatores de risco relacionados entre os escolares do ensino fundamental I e II na cidade de João Pessoa, no nordeste do Brasil

The goal of the present study was to determine both prevalence and risk factors associated with intestinal parasitism in school students. A cross-sectional study was conducted in a single primary school located in João Pessoa, from February to August in 2016. The students were selected from the age group of 5-16 years. Of the school total of 341 students, 150 fecal specimens (from participants) were collected and were evaluated by three methods: Hoffman, Pons, and Janer (HPJ); Rugai; and the Paratest® Kit. A questionnaire concerning socio-demographic, environmental and behavioral variables was also applied. A logistic regression model was used to explain the occurrence of intestinal parasitism and the associated risk factors. The prevalence was 38.7% of students, with positive samples being more prevalent in the male students (47.0%). The most common parasite was Giardia lamblia 13 (14.8%), followed by Entamoeba histolytica/dispar 8 (9%), Enterobius vermicularis 5 (5.7%), Strongyloides stercolaris 2 (2.3%), Ascaris lumbricoides 2 (2.3%) and Trichuris trichiura 2 (2.3%). Among the enterocommensals, the most frequent was Endolimax nana 36 (40.9%) followed by Entamoeba coli 20 (22.7%). The variables that presented statistical significance (pvalue< 0.05) ) together with the Odds Ratio (OR) were: gender (female) (OR=2.4; 95% CI, 0.19-0.98), family allowance participant (yes) (OR=4.4; 95% CI, 1.84-10.66), number of rooms in the residence (OR=3.5; 95% CI, 1.13-10.64), family nucleus (OR=7.0; 95% CI, 1.46-12.43), fruit and vegeTable hygiene (OR=2.0; 95% CI, 1.23-3.36), use of anthelmintic (OR= 0.02; 95% CI, 0.001-0.30) and detection of worms (OR=25.0; 95% CI, 20.6-30.10). Diseases caused by protozoa were more prevalent. The analyzed risk factors demonstrate that disease transmission happens through differing routes. Thus, appropriate health intervention strategies should be implemented to reduce the burden of intestinal parasites for school students and their families.

O presente estudo tem como objetivo determinar a prevalência e os fatores de risco associados ao parasitismo intestinal em escolares. Este estudo transversal foi conduzido em uma escola primária localizada na cidade de João Pessoa, de fevereiro a agosto de 2016. As crianças foram selecionadas entre 5 a 16 anos de idade. De 341 alunos, foram coletados 150 espécimes fecais e foram avaliados por três métodos: Método de Hoffman, Pons e Janer (HPJ); Kit Rugai e Paratest®. Foi preenchido um questionário sobre dados de variáveis sociodemográficas, ambientais e de comportamento. O modelo de regressão logística foi utilizado para explicar a ocorrência de parasitismo intestinal e os fatores de risco associados. A prevalência foi de 38,7% das crianças, sendo a amostra positiva mais predominante nas crianças do sexo masculino (47,0%). O parasita mais comum foi Giardia lamblia 13 (14,8%), seguido de Entamoeba histolytica / dispar 8 (9%), Enterobius vermicularis 5 (5,7%), Strongyloides stercolaris 2 (2,3%), Ascaris lumbricoides 2 (2,3%) e Trichuris Trichiura 2 (2,3%). Entre os enterocomensais, a maioria das freqüências foi Endolimax nana 36 (40,9%) seguido de Entamoeba coli 20 (22,7%). As variáveis que apresentaram significância estatística (p-valor <0,05) foram: gênero (feminino) (OR = 2,4; IC 95%; 0,19-0,98), recebe subsídio familiar (sim) (OR = 4,4; 95% CI, 1,84-10,66), número de quartos na residência (OR = 3,5; IC 95%, 1,13-10,64), núcleo familiar (OR = 7,0; IC 95%; 1,46- 12,43), higiene das frutas e legumes (OR = 2,0, IC 95%, 1,23-3,36), uso de vermifugio (OR = 0,02, IC 95%, 0,001-0,30) e visualização de vermes (OR = 25,0; IC 95%, 20,6-30,10). As doenças causadas por protozoários foram mais prevalentes. Os fatores de risco analisados demonstram que a transmissão de doenças ocorre por rotas diferentes. Portanto, as estratégias de intervenção em saúde devem ser implementadas para as crianças da escola e suas famílias para reduzir o peso dos parasitas intestinais.

High prevalence of iodine deficiency in pregnant women living in adequate iodine area.

OBJECTIVES: Iodine deficiency during pregnancy is associated with obstetric and neonatal adverse outcomes. Serum thyroglobulin (sTg) and thyroid volume (TV) are optional tools to urinary iodine concentration (UIC) for defining iodine status. This cross-sectional study aims to evaluate the iodine status of pregnant women living in iodine-adequate area by spot UIC and correlation with sTg, TV and thyroid function. METHODS: Two hundred and seventy-three pregnant women were evaluated at three trimesters. All had no previous thyroid disease, no iodine supplementation and negative thyroperoxidase and thyroglobulin antibodies. Thyroid function and sTg were measured using electrochemiluminescence immunoassays. TV was determined by ultrasonography; UIC was determined using a modified Sandell-Kolthoff method. RESULTS: Median UIC was 146 µg/L, being 52% iodine deficient and only 4% excessive. TSH values were 1.50 ± 0.92, 1.50 ± 0.92 and 1.91 ± 0.96 mIU/L, respectively, in each trimester (P = 0.001). sTg did not change significantly during trimesters with median 11.2 ng/mL and only 3.3% had above 40 ng/mL. Mean TV was 9.3 ± 3.4 mL, which positively correlated with body mass index, but not with sTg. Only 4.5% presented with goitre.When pregnant women were categorized as iodine deficient (UIC < 150 µg/L), adequate (≥150 and <250 µg/L) and excessive (≥250 µg/L), sTg, thyroid hormones and TV at each trimester showed no statistical differences. CONCLUSIONS: Iodine deficiency was detected frequently in pregnant women living in iodine-adequate area. sTg concentration and TV did not correlate to UIC. Our observation also demonstrated that the Brazilian salt-iodization programme prevents deficiency, but does not maintain iodine status within adequate and recommended ranges for pregnant women.

Neurogenic bladder findings in patients with Congenital Zika Syndrome: A novel condition.

INTRODUCTION: Congenital Zika Syndrome (CZS) has been associated with microcephaly and other central nervous system abnormalities including areas that have been implicated in the control of the lower urinary tract. As such, this descriptive case series has aimed to investigate whether CZS is linked with neurogenic bladder. Identifying such an association is paramount in the effort to recognize CZS complications that have putative treatment options that could mitigate the impact of CZS in infected children. METHODS: Following IRB approval, urological assessment was performed in all patients referred to our clinic between June 2016 and May 2017 who presented with confirmed CZS-associated microcephaly. The research protocol consisted of obtaining clinical history, laboratory tests, lower and upper urinary tract ultrasounds, as well as a diagnostic urodynamic evaluation. ZIKA virus infection was previously confirmed by maternal history and positive PCR in babies and mothers. Microcephaly and other central nervous system abnormalities were established based on neurological assessment and associated imaging of the central nervous system (CT head and/or Brain MRI). RESULTS: Twenty-two consecutive CZS patients were tested and confirmed to have neurogenic bladder. Of the 22 patients assessed, 21 presented with an overactive bladder combined with reduced bladder capacity and elevated detrusor filling pressures. Clinically significant increases in postvoid residual (PVR) were confirmed in 40% of cases while a urinary tract infection (UTI) was identified in 23% of cases. CONCLUSION: Neurogenic bladder, a known treatable health condition, was confirmed in 100% of patients tested in this study, most presenting with high-risk urodynamic patterns known to lead to renal damage when left untreated. Follow up studies are necessary to provide further insight onto long-term disease progression and to investigate the response to standard therapies for neurogenic bladder. Nonetheless, we emphasize the importance of proactive management of neurogenic bladder and prompt referral so as to help mitigate CZS disease burden for patients and their families.

Efficient method for comprehensive computation of agent-level epidemic dissemination in networks.

Susceptible-infected (SI) and susceptible-infected-susceptible (SIS) are simple agent-based models often employed in epidemic studies. Both models describe the time evolution of infectious diseases in networks whose vertices are either susceptible (S) or infected (I) agents. Precise estimation for disease spreading is one of the major goals in epidemic studies but often restricted to heavy numerical simulations. Analytic methods using operatorial content are subject to the asymmetric eigenvalue problem, limiting the use of perturbative methods. Numerical methods are limited to small populations, since the vector space increases exponentially with population size N. Here, we propose the use of the squared norm of the probability vector to obtain an algebraic equation, which permits the evaluation of stationary states in Markov processes. The equation requires the eigenvalues of symmetrized time generators and takes full advantage of symmetries, reducing the time evolution to an O(N) sparse problem. The calculation of eigenvalues employs quantum many-body techniques, while the standard perturbation theory accounts for small modifications to the network topology.

Effects of cytoskeletal drugs on actin cortex elasticity.

Mechanical properties of cells are known to be influenced by the actin cytoskeleton. In this article, the action of drugs that interact with the actin cortex is investigated by tether extraction and rheology experiments using optical tweezers. The influences of Blebbistatin, Cytochalasin D and Jasplakinolide on the cell mechanical properties are evaluated. The results, in contradiction to current views for Jasplakinolide, show that all three drugs and treatments destabilize the actin cytoskeleton, decreasing the cell membrane tension. The cell membrane bending modulus increased when the actin cytoskeleton was disorganized by Cytochalasin D. This effect was not observed for Blebbistatin and Jasplakinolide. All drugs decreased by two-fold the cell viscoelastic moduli, but only Cytochalasin D was able to alter the actin network into a more fluid-like structure. The results can be interpreted as the interplay between the actin network and the distribution of myosins as actin cross-linkers in the cytoskeleton. This information may contribute to a better understanding of how the membrane and cytoskeleton are involved in cell mechanical properties, underlining the role that each one plays in these properties.

A T Cell View of the Bone Marrow.

The majority of T cells present in the bone marrow (BM) represent an activated/memory phenotype and most of these, if not all, are circulating T cells. Their lodging in the BM keeps them activated, turning the BM microenvironment into a "memory reservoir." This article will focus on how T cell activation in the BM results in both direct and indirect effects on the hematopoiesis. The hematopoietic stem cell niche will be presented, with its main components and organization, along with the role played by T lymphocytes in basal and pathologic conditions and their effect on the bone remodeling process. Also discussed herein will be how "normal" bone mass peak is achieved only in the presence of an intact adaptive immune system, with T and B cells playing critical roles in this process. Our main hypothesis is that the partnership between T cells and cells of the BM microenvironment orchestrates numerous processes regulating immunity, hematopoiesis, and bone remodeling.

Effect of stocking density and genetic group on mineral composition and development of long bones of broilers / Efeito da densidade de criação e do grupo genético sobre a composição mineral e desenvolvimento de ossos longos de frangos de corte

The objective of this experiment was to evaluate the mineral composition, volume, bone breaking strenght and optical radiographic density of long bones (humerus, tibia and fêmur) of broiler chickens from three genetic groups and two rearing densities. The experimental design was a splitj-splot where the main plots were a combination of three genetic groups (Isa Label JA57, Ross 308 and Hybro PG) and two rearing densities ( 10 and 16 birds/m2) in a factorial arrangement, and age was considered a secondary plot, so there was six treatments with 5 replications per each treatment, totalizing 30 experimental units. The results founded showed that there were no differences in the studied genetic groups when submitted to two rearing densities for all evaluated parameters, with exception for bone breaking strength of tibia from Isa Label JA57. Bone volume increased with broiler age, while bone breaking strength and bone density showed a decreased from 28 to 35 days of age. Bone mineral content for all treatments showed up growing in the first three weeks, decreasing in the end of the experimental period. The pattern of bone development was the same for all genetic groups, despite the rearing density used, showing that broilers had a standard bone growth curve that is not affected by genetic or rearing density.(AU)

O objetivo do trabalho foi avaliar a composição mineral, volume, resistência e densidade óptica radiográfica dos ossos longos (úmero, tíbia e fêmur) de frangos de corte de três grupos genéticos e duas densidades de alojamento. O delineamento experimental utilizado foi inteiramente casualizado em um esquema fatorial 3 x 2, em parcelas subdivididas, sendo os fatores principais os três grupos genéticos (Isa Label JA57, Ross 308 e Hybro PG) e as duas densidades de alojamento (10 e 16 aves/ m2), e as subparcelas as idades de coleta dos ossos (7 semanas), totalizando deste modo seis tratamentos com cinco repetições cada um num total de 30 unidades experimentais. Os resultados deste trabalho apontaram que não houve diferenças significativas para as características avaliadas com exceção da resistência da tíbia para o grupo Isa Label JA57. O volume ósseo apresentou-se crescente com a idade da ave; já a resistência à quebra e densidade óptica radiográfica para a maioria dos tratamentos avaliados decresceram dos 28 aos 35 dias. O teor de minerais dos ossos avaliados para todos os tratamentos mostrou-se crescente nas três primeiras semanas, decrescendo no final do período experimental. O modelo de desenvolvimento ósseo foi o mesmo para todos os grupos genéticos, apesar da densidade de criação adotada, mostrando que os frangos de corte possuem uma curva padrão de crescimento ósseo, que não é afetada pelo grupo genético ou pela densidade de criação.(AU)

Effect of stocking density and genetic group on mineral composition and development of long bones of broilers / Efeito da densidade de criação e do grupo genético sobre a composição mineral e desenvolvimento de ossos longos de frangos de corte

The objective of this experiment was to evaluate the mineral composition, volume, bone breaking strenght and optical radiographic density of long bones (humerus, tibia and fêmur) of broiler chickens from three genetic groups and two rearing densities. The experimental design was a splitj-splot where the main plots were a combination of three genetic groups (Isa Label JA57, Ross 308 and Hybro PG) and two rearing densities ( 10 and 16 birds/m2) in a factorial arrangement, and age was considered a secondary plot, so there was six treatments with 5 replications per each treatment, totalizing 30 experimental units. The results founded showed that there were no differences in the studied genetic groups when submitted to two rearing densities for all evaluated parameters, with exception for bone breaking strength of tibia from Isa Label JA57. Bone volume increased with broiler age, while bone breaking strength and bone density showed a decreased from 28 to 35 days of age. Bone mineral content for all treatments showed up growing in the first three weeks, decreasing in the end of the experimental period. The pattern of bone development was the same for all genetic groups, despite the rearing density used, showing that broilers had a standard bone growth curve that is not affected by genetic or rearing density.

O objetivo do trabalho foi avaliar a composição mineral, volume, resistência e densidade óptica radiográfica dos ossos longos (úmero, tíbia e fêmur) de frangos de corte de três grupos genéticos e duas densidades de alojamento. O delineamento experimental utilizado foi inteiramente casualizado em um esquema fatorial 3 x 2, em parcelas subdivididas, sendo os fatores principais os três grupos genéticos (Isa Label JA57, Ross 308 e Hybro PG) e as duas densidades de alojamento (10 e 16 aves/ m2), e as subparcelas as idades de coleta dos ossos (7 semanas), totalizando deste modo seis tratamentos com cinco repetições cada um num total de 30 unidades experimentais. Os resultados deste trabalho apontaram que não houve diferenças significativas para as características avaliadas com exceção da resistência da tíbia para o grupo Isa Label JA57. O volume ósseo apresentou-se crescente com a idade da ave; já a resistência à quebra e densidade óptica radiográfica para a maioria dos tratamentos avaliados decresceram dos 28 aos 35 dias. O teor de minerais dos ossos avaliados para todos os tratamentos mostrou-se crescente nas três primeiras semanas, decrescendo no final do período experimental. O modelo de desenvolvimento ósseo foi o mesmo para todos os grupos genéticos, apesar da densidade de criação adotada, mostrando que os frangos de corte possuem uma curva padrão de crescimento ósseo, que não é afetada pelo grupo genético ou pela densidade de criação.

T cells induce pre-metastatic osteolytic disease and help bone metastases establishment in a mouse model of metastatic breast cancer.

Bone metastases, present in 70% of patients with metastatic breast cancer, lead to skeletal disease, fractures and intense pain, which are all believed to be mediated by tumor cells. Engraftment of tumor cells is supposed to be preceded by changes in the target tissue to create a permissive microenvironment, the pre-metastatic niche, for the establishment of the metastatic foci. In bone metastatic niche, metastatic cells stimulate bone consumption resulting in the release of growth factors that feed the tumor, establishing a vicious cycle between the bone remodeling system and the tumor itself. Yet, how the pre-metastatic niches arise in the bone tissue remains unclear. Here we show that tumor-specific T cells induce osteolytic bone disease before bone colonization. T cells pro-metastatic activity correlate with a pro-osteoclastogenic cytokine profile, including RANKL, a master regulator of osteoclastogenesis. In vivo inhibition of RANKL from tumor-specific T cells completely blocks bone loss and metastasis. Our results unveil an unexpected role for RANKL-derived from T cells in setting the pre-metastatic niche and promoting tumor spread. We believe this information can bring new possibilities for the development of prognostic and therapeutic tools based on modulation of T cell activity for prevention and treatment of bone metastasis.

Increased bone loss and amount of osteoclasts in kinin B1 receptor knockout mice.

AIM: The pathophysiology of periodontal diseases involves aspects of immunity and bone remodelling. Considering the role of the kinin B1 receptor (Bdkrb1) in inflammation and healing, the purpose of this study was to evaluate the contribution of Bdkrb1 to the pathogenesis of periodontitis. MATERIAL AND METHODS: We used a model of ligature-induced experimental periodontitis (LIEP) in mice lacking Bdkrb1 (Bdkrb1(-/-) ) to test the role of this receptor in bone loss and cytokine secretion by lymph nodes cells. Angiotensin-converting enzyme inhibitor (ACEi) was used as a pharmacological strategy to support the genetic model. Also, autonomous effect of Bdkrb1 deletion was evaluated in osteoclasts precursors from bone marrow. RESULTS: Bdkrb1(-/-) mice exhibit increased bone loss and IL-17 secretion in response to LIEP when compared to wild type. LIEP does not modify TNF-α, IFN-γ and IL-10 levels in Bdkrb1(-/-) mice after 21 days. Bone marrow cells from Bdkrb1(-/-) displayed increased differentiation into functional osteoclasts with consistent artificial calcium phosphate degradation. Furthermore, treatment of mice with ACEi prevented bone destruction. CONCLUSION: Bdkrb1 participates in the pathogenesis of LIEP bone loss possibly through mechanisms that involve modulation of the TH 17 response, thereby demonstrating its role in the development of periodontitis.

Dehydrated citrus pulp digestibility and the effect of its inclusion in diet on the performance of growing rabbits / Digestibilidade da polpa cítrica desidratada e efeito de sua inclusão na dieta sobre o desempenho de coelhos em crescimento

Several experiments were conducted to determine the digestibility of dehydrated citrus pulp (DCP) and evaluate its effect on rabbit feed. Digestibility assay comprised 30 rabbits fed on two diets: a reference diet and a diet in which 20% consisted of DCP. The digestibility assay comprised 10 days for adaptation and four days for feces collection. The digestibility coefficient and digestible nutrient rates for DM, CP, NDF, ADF, and CE of DCP were respectively 83.75 and 76.04%; 70.12 and 5.03%; 40.14 and 9.52%; 68.98 and 15.09%; and 81.48% and 3394 kcalDE kg-

Conduziram-se experimentos para determinar a digestibilidade da polpa cítrica desidratada (PCD) e avaliá-la na alimentação de coelhos. Para o ensaio de digestibilidade utilizaram-se 30 coelhos recebendo duas dietas: uma referência e outra com substituição de 20% da primeira por PCD. O ensaio teve duração de dez dias para adaptação mais quatro para coleta de fezes. Os coeficientes de digestibilidade e os nutrientes digestíveis para MS, PB, FDN, FDA e EB da PCD foram respectivamente: 83,75 e 76,04%; 70,12 e 5,03%; 40,14 e 9,52%; 68,98 e 15,09%; e 81,48% e 3394 kcalED kg

Dehydrated citrus pulp digestibility and the effect of its inclusion in diet on the performance of growing rabbits / Digestibilidade da polpa cítrica desidratada e efeito de sua inclusão na dieta sobre o desempenho de coelhos em crescimento

Several experiments were conducted to determine the digestibility of dehydrated citrus pulp (DCP) and evaluate its effect on rabbit feed. Digestibility assay comprised 30 rabbits fed on two diets: a reference diet and a diet in which 20% consisted of DCP. The digestibility assay comprised 10 days for adaptation and four days for feces collection. The digestibility coefficient and digestible nutrient rates for DM, CP, NDF, ADF, and CE of DCP were respectively 83.75 and 76.04%; 70.12 and 5.03%; 40.14 and 9.52%; 68.98 and 15.09%; and 81.48% and 3394 kcalDE kg-

Conduziram-se experimentos para determinar a digestibilidade da polpa cítrica desidratada (PCD) e avaliá-la na alimentação de coelhos. Para o ensaio de digestibilidade utilizaram-se 30 coelhos recebendo duas dietas: uma referência e outra com substituição de 20% da primeira por PCD. O ensaio teve duração de dez dias para adaptação mais quatro para coleta de fezes. Os coeficientes de digestibilidade e os nutrientes digestíveis para MS, PB, FDN, FDA e EB da PCD foram respectivamente: 83,75 e 76,04%; 70,12 e 5,03%; 40,14 e 9,52%; 68,98 e 15,09%; e 81,48% e 3394 kcalED kg <->1 MS. Para o desempenho utilizaram-se 120 coelhos recebendo dietas com diferentes níveis de PCD (0, 20, 40, 60, 80 e 100%) substituindo o milho. Observou-se redução linear (p < 0,01) para o consumo de ração e ganho de peso diário, peso vivo aos 50 e 70 dias e rendimento de carcaça com o aumento gradativo de PCD. Efeitos quadráticos (p < 0,01) sobre a conversão alimentar dos 32 aos 50 e 32 aos 70 dias de idade provocaram melhores valores com 42,74 e 44,40% de PCD, respectivamente. PCD a 20% não afetou (p > 0,05) nenhuma característica de desempenho quando comparada à dieta controle. Conclui-se: a PCD pode substituir o milho até 20% nas dietas de coelhos.(AU)

Mining gene expression signature for the detection of pre-malignant melanocytes and early melanomas with risk for metastasis.

BACKGROUND: Metastatic melanoma is a highly aggressive skin cancer and currently resistant to systemic therapy. Melanomas may involve genetic, epigenetic and metabolic abnormalities. Evidence is emerging that epigenetic changes might play a significant role in tumor cell plasticity and metastatic phenotype of melanoma cells. PRINCIPAL FINDINGS: In this study, we developed a systematic approach to identify genes implicated in melanoma progression. To do this, we used the Affymetrix GeneChip Arrays to screen 34,000 mouse transcripts in melan-a melanocytes, 4C pre-malignant melanocytes, 4C11- non-metastatic and 4C11+ metastatic melanoma cell lines. The genome-wide association studies revealed pathways commonly over-represented in the transition from immortalized to pre-malignant stage, and under-represented in the transition from non-metastatic to metastatic stage. Additionally, the treatment of cells with 10 µM 5-aza-2'-deoxycytidine (5AzaCdR) for 48 hours allowed us to identify genes differentially re-expressed at specific stages of melan-a malignant transformation. Treatment of human primary melanocytes with the demethylating agent 5AzaCdR in combination to the histone deacetylase inhibitor Trichostatin A (TSA) revealed changes on melanocyte morphology and gene expression which could be an indicator of epigenetic flexibility in normal melanocytes. Moreover, changes on gene expression recognized by affecting the melanocyte biology (NDRG2 and VDR), phenotype of metastatic melanoma cells (HSPB1 and SERPINE1) and response to cancer therapy (CTCF, NSD1 and SRC) were found when Mel-2 and/or Mel-3-derived patient metastases were exposed to 5AzaCdR plus TSA treatment. Hierarchical clustering and network analyses in a panel of five patient-derived metastatic melanoma cells showed gene interactions that have never been described in melanomas. SIGNIFICANCE: Despite the heterogeneity observed in melanomas, this study demonstrates the utility of our murine melanoma progression model to identify molecular markers commonly perturbed in metastasis. Additionally, the novel gene expression signature identified here may be useful in the future into a model more closely related to translational research.

Efeito de dietas semi-simplificadas formuladas com subprodutos demandioca ensilados ou não sobre o desempenho e característicasde carcaça de coelhos / Effect of half-simplified diets formulated with cassava by-product ensiled or not on performance and carcass characteristics of rabbits

Foram avaliados o desempenho da desmama ao abate e as características de carcaça de 144 coelhos da raça Nova Zelândia Branco, oriundos de matrizes alimentadas com dietas referência ou semi-simplificada com 79,83% de subprodutos de mandioca, distribuídos em gaiolas, em esquema fatorial 2 x 3 (duas rações fornecidas às matrizes e láparos até a desmama versus três rações fornecidas aos animais da desmama até o abate) com 12 repetições e dois animais por unidade experimental. As dietas fornecidas, após a desmama, consistiram de uma referência e duas semi-simplificadas com 81,63% de subprodutos de mandioca ensilados ou não antes da sua incorporação. Coelhos cujas mães foram alimentadas com dieta semi-simplificadas se apresentaram mais leves à desmama que os desmamados com dieta-referência. Observou-se menor consumo de ração para os coelhos que receberam a dieta maternidade semi-simplificada até a desmama, resultando em melhor conversão alimentar. Os pesos da carcaça e dos cortes comerciais foram maiores nos animais alimentados com as rações referência maternidade e referência crescimento. O uso de dietas semi-simplificadas com subprodutos de mandioca ensilados ou não, da desmama ao abate, reduziu o ganho de peso diário e o consumo de ração, porém melhorou a conversão alimentar e diminuiu o custo de ração. A utilização das rações semi-simplificadas apresenta-se como alternativa interessante para a produção de coelhos.

This study evaluated the performance from weaning to slaughter and carcass characteristics of 144 New Zealand White rabbits, from does fed with a reference diet or half-simplified diet containing 79.83% cassava by-product, allocated in iron cages, in a 2 x 3 factorial arrangement (two diets used for does and litters until weaning versus three diets supplied for animals from weaning to slaughter) with 12 replications and two animals per experimental unit. The supplied diets after weaning consisted of one reference and two half-simplified diets with 81.63% cassava by-product ensiled or not before its incorporation. Rabbits from mothers fed with the half-simplified diet gained less weight at weaning than rabbits fed with reference diet. Lower feed intake was observed for rabbits that received the maternity half-simplified diet until weaning, resulting in a better feed:gain ratio. The weight of carcass and commercial cuts were higher for animals fed with both reference diets: maternity and growth diets. The use of half-simplified diets with cassava by-product ensiled or not, from weaning to slaughter, reduced daily weight gain and feed intake; however, it improved the feed:gain ratio and decreased ration feed cost. The use of halfsimplified diets represents an interesting alternative in rabbit production.