Neutrophil Granulocytes in Ovarian Cancer - Induction of Epithelial-To- Mesenchymal-Transition and Tumor Cell Migration

Background: Ovarian cancer (OvCa) is a highly aggressive malignoma with a tumor-promoting microenvironment. Infiltration of polymorphonuclear neutrophils (PMN) is frequently seen, raising the question of their impact on tumor development. In that context, effects of PMN on human ovarian cancer cells were assessed. Methods: Human epithelial ovarian cancer cells were incubated with human PMN, lysate of PMN, or neutrophil elastase. Morphological alterations were observed by time-lapse video-microscopy, and the underlying molecular mechanism was analyzed by flow cytometry and Western blotting. Functional alternations were assessed by an in vitro wound healing assay. In parallel, a large cohort of n=334 primary OvCa tissue samples of various histological subtypes was histologically evaluated. Results: Co-cultivation of cancer cells with either PMN or PMN lysate causes a change of the polygonal epithelial phenotype of the cells towards a spindle shaped morphology, causing a cribriform cell growth. The PMN-induced alteration could be attributed to elastase, a major protease of PMN. Elastase-induced shape change was most likely due to the degradation of membranous E-cadherin, which results in loss of cell contacts and polarity. Moreover, in response to elastase, epithelial cytokeratins were downmodulated, in parallel with a nuclear translocation of β-catenin. These PMN-elastase induced alterations of cells are compatible with an epithelial-to-mesenchymal transition (EMT) of the cancer cells. Following EMT, the cells displayed a more migratory phenotype. In human biopsies, neutrophil infiltration was seen in 72% of the cases. PMN infiltrates were detected preferentially in areas with low E-cadherin expression. Conclusion: PMN in the microenvironment of OvCa can alter tumor cells towards a mesenchymal and migratory phenotype

AVALIAÇÃO DO USO DE SANGUE EM PAPEL-FILTRO PARA DETECÇÃO E QUANTIFICAÇÃO DE ANTICORPOS PARA O VÍRUS DA DOENÇA DE NEWCASTLE

O presente trabalho descreve a utilização de sangue total em papel-filtro para avaliação dos níveis de anticorpos específicos contra o vírus da doença de Newcastle (NDV) através da técnica de inibição da hemaglutinação (HI). Coletaram-se e analisaram-se por HI amostras de sangue de noventa e oito galinhas, com um histórico conhecido de vacinação contra o NDV HI. O sangue foi impregnado em papel-filtro Wathman n° 1 com 3cm2 e também coletado em tubos para a obtenção de soro. Armazenaram-se os papéis à temperatura ambiente e procedeu-se a sua análise num período de até quarenta e cinco dias. A sensibilidade, a es¬pecificidade, o valor preditivo positivo e o valor preditivo negativo do HI mediante o uso de papel-filtro, comparado com o HI em que se utilizou soro, foram de 89%, 73%, 76% e 87 %, respectivamente. Detectaram-se um índice de pre¬cisão de 80% e um coeficiente de correlação r = 0,81%. O título médio geométrico de anticorpos foi 33,1% e 33,6 para eluídos de papel-filtro e os soros, respectivamente. A asso¬ciação entre os dois métodos foi altamente significativa (p< 0.001) quando analisada pelo teste de Fisher. Os resultados indicam o potencial da utilização do sangue em papel-filtro como uma alternativa ao soro para detecção de anticorpos contra o NDV. PALAVRAS-CHAVES: Galinha, inibição da hemaglutinação, sorologia

Efeito do extrato etanólico de própolis verde sobre a produção de anticorpos após imunização contra parvovírus canino (CPV) e coronavírus canino (CCoV)

Este estudo foi realizado para avaliar se extrato etanólico de própolis verde (EEP) pode interferir na produção de anticorpos específicos após imunização contra parvovírus (CPV) e coronavírus canino (CCoV). Camundongos foram vacinados com CPV e CCoV (0.75, 1.5 e 3 x 106 TCID50) com ou sem 400 μg/dose de EEP. Vinte e um dias após a terceira dose foi mensurado IgG sérica. A coadministração de EEP aumentou significativamente os níveis de IgG específica para o CPV em animais inoculados com a maior concentração do antígeno, e não teve influência sobre os níveis de anticorpos para CCoV. Os resultados indicam que o EEP tem ação imunomoduladora intimamente dependente do tipo e concentração do antígeno utilizado, sendo capaz de aumentar os níveis de anticorpos contra CPV.This study was designed to evaluate whether an ethanolic extract of green propolis (EEP) can interfere with production of specific antibodies after immunization against parvovirus (CPV) and canine coronavirus (CCoV). Mice were vaccinated with CPV and CCoV (0.75, 1.5 and 3 x 106 TCID50) with or without 400 μg/dose of the EEP. Twenty one days after the third dose was measured serum IgG. The co-administration of the EEP significantly enhanced serum specific IgG responses to CPV in animals inoculated with the highest concentration of the antigen, and had no influence on levels of antibodies to CCoV. The results indicate that the EEP has immunomodulatory action closely dependent on the type and concentration of antigen used, being able to increase the levels of antibodies to CPV

Risk Factors Associated with the Presence of Viral Diseases in Domestic Poultry in the Southern Region of Rio Grande do Sul, Brazil

Background: The introduction of any infectious agent into an industrial or subsistence farm worries agribusiness owners in Brazil because it reduces product quality and increases treatment costs, although most diseases are untreatable, thus causing economic losses with morbidity and mortality. Therefore, an epidemiological survey of viral diseases associated with poultry was developed by performing a detailed description of the risk factors that may be related to existing diseases using domestic poultry sample data recorded in the Regional Diagnostic Laboratory (LRD) of College of Veterinary Medicine of the Federal University of Pelotas (UFPel), Rio Grande do Sul, Brazil, from 2000 to 2016.Materials, Methods & Results: Epidemiological and clinical-pathological data were collected and then compared with disease data by multivariate analysis using statistical EpiInfo version 6.04 and Microsoft Office Excel 2010 software. The frequencies and 95% confidence intervals (CI), association measures (odds ratio=OR and relative risk=RR), Chi-square test, and the results considered significant with a value of P ≤ 0.05 were described. A total of 410 samples of domestic poultry were tested, and the results showed 66 (16.1%) viral diseases. The following conditions were the most commonly found diseases in this study: Marek’s disease (42.4%), Infectious bursal disease (31.8%), Avian leukosis (16.6%), Avian pox (7.5%) and Avian infectious bronchitis (1.5%). In this article we discuss the most frequent viral diseases: Marek’s disease (DM) and Gumboro disease. It was also possible to conclude that birds with Marek's disease presented higher odds of developing nerve, tegumentary and locomotors signs (P ≤ 0.05).  As well as, more likely to present tumoriform lesions in the liver, spleen, kidneys and heart P ≤ 0.05, as well as lesions in the proventriculus, muscle lesions and in the sciatic nerve P ≤ 0.05. Laying poultry, backyard poultry production and young poultry constitute risk factors for DM (P ≤ 0.05).  The results showed that samples with mycotoxicoses were at higher risk of developing DM (P ≤ 0.05). Poultry from industrial system had a higher risk of developing Gumboro disease (P ≤ 0.05). Besides that, domestic poultry with Gumboro disease were more likely to develop signs of depression, nerve signals, tegumentary changes, bursitis, hepatomegaly, renal lesions and splenomegaly (P ≤ 0.05).Discussion: In this research we can observe the domestic poultry with DM presenting tumoriform lesions in several organs, this finding is common in this disease, because it is one of the most frequent viral diseases of neoplastic characteristic. The results show that young birds are a risk factor for DM. In fact, previous works already affirm young birds as being susceptible to disease in relation to adult birds. In this study, laying hens presented a risk factor for DM, however other authors also observed in cut birds or in both types of production. Industrial poultry are at higher risk for DM, this was also reported by other authors, because the disease is of high contagion and morbidity, industrial birds are under high density which facilitates the contact and dissemination of the virus among sick and susceptible birds. In this study, industrial poultry were a risk factor for Gumboro disease, however, other studies show that there is no difference between types of breeding and can also occur in backyard birds. Young poultry and laying hens are more susceptible, and the clinical signs found in the study are compatible with those described in the literature. Through this research on viral diseases, we conclude that our study may complement the already existing official data, since, in a research conducted in the same period, it did not identify the presence of viral diseases in the Southern of RS, but with our research it was possible to register this disease

Chemical characterization, antioxidant, cytotoxic, and antibacterial activities of Eugenia uniflora L. and Psidium cattleianum Sabine essential oils against Klebsiella pneumoniae and Acinetobacter baumannii/ Caracterização química, antioxidante, citotóxica e antibacteriana dos óleos essenciais de Eugenia uniflora L. e Psidium cattleianum Sabine contra Klebsiella pneumoniae e Acinetobacter baumannii

INTRODUCTION: Eugenia uniflora L. and Psidium cattleianum Sabine essential oils (EO) can be biologically active and serve as novel sources of antibiotics for Klebsiella pneumoniae and Acinetobacter baumannii bacteria.METHODS: The EO of E. uniflora L. (EOE) and P. cattleianum Sabine (EOP) were extracted from de leaves of the plants and chemically characterized by gas chromatography-mass spectrometry (GC-MS). The antioxidant potential was evaluated by the DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) technique and the cytotoxicity evaluated in mammalian VERO cell line by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The antibacterial activity of EOE and EOP was assessed by the broth microdilution method.RESULTS: The major compounds of EOE were benzofuran (24.38%), germacrene B (20.12%), ?-elemene (9.33%) and ?-cubebene (8.55%), and for EOP were ?-pinene (24.25%), ?-caryophyllene (20.45%), and eucalyptol (10.43%). Both EO tested showed low antioxidant effect and dose-dependent cytotoxicity in VERO cell line. The EOE was less toxic for the mammalian cells with an IC50 (half maximal inhibitory concentration) index of 75.0 mg.mL-1. The EOP and EOE were more active against A. baumannii, with a minimum inhibitory concentration (MIC) of 14.0 and 56.0 mg.mL-1 and a minimal bactericide concentration (MBC) of 14.0 and 112.0 mg.mL-1, respectively.  CONCLUSIONS: Although EOE and EOP have low antioxidant and antibacterial activity against A. baumanni and K. pneumoniae new approaches can be applied to improve this effect in vivo.  

Glycaemic control and antidiabetic therapy in patients with diabetes mellitus and chronic kidney disease - cross-sectional data from the German Chronic Kidney Disease (GCKD) cohort

Background: Diabetes mellitus (DM) is the leading cause of end-stage renal disease. Little is known about practice patterns of anti-diabetic therapy in the presence of chronic kidney disease (CKD) and correlates with glycaemic control. We therefore aimed to analyze current antidiabetic treatment and correlates of metabolic control in a large contemporary prospective cohort of patients with diabetes and CKD. Methods: The German Chronic Kidney Disease (GCKD) study enrolled 5217 patients aged 18-74 years with an estimated glomerular filtration rate (eGFR) between 30-60 mL/min/1.73 m(2) or proteinuria >0.5 g/d. The use of diet prescription, oral anti-diabetic medication, and insulin was assessed at baseline. HbA1c, measured centrally, was the main outcome measure. Results: At baseline, DM was present in 1842 patients (35 %) and the median HbA1C was 7.0 % (25th-75th percentile: 6.8-7.9 %), equalling 53 mmol/mol (51, 63);24.2 % of patients received dietary treatment only, 25.5 % oral antidiabetic drugs but not insulin, 8.4 % oral antidiabetic drugs with insulin, and 41.8 % insulin alone. Metformin was used by 18.8 %. Factors associated with an HbA1C level >7.0 % (53 mmol/mol) were higher BMI (OR = 1.04 per increase of 1 kg/m(2), 95 % CI 1.02-1.06), hemoglobin (OR = 1.11 per increase of 1 g/dL, 95 % CI 1.04-1.18), treatment with insulin alone (OR = 5.63, 95 % CI 4.26-7.45) or in combination with oral antidiabetic agents (OR = 4.23, 95 % CI 2.77-6.46) but not monotherapy with metformin, DPP-4 inhibitors, or glinides. Conclusions: Within the GCKD cohort of patients with CKD stage 3 or overt proteinuria, antidiabetic treatment patterns were highly variable with a remarkably high proportion of more than 50 % receiving insulin-based therapies. Metabolic control was overall satisfactory, but insulin use was associated with higher HbA1C levels

Blood pressure control in chronic kidney disease: A cross-sectional analysis from the German Chronic Kidney Disease (GCKD) study

We assessed the prevalence, awareness, treatment and control of hypertension in patients with moderate chronic kidney disease (CKD) under nephrological care in Germany. In the German Chronic Kidney Disease (GCKD) study, 5217 patients under nephrology specialist care were enrolled from 2010 to 2012 in a prospective observational cohort study. Inclusion criteria were an estimated glomerular filtration rate (eGFR) of 30 +/- 60 mL/min/1.73 m 2 or overt proteinuria in the presence of an eGFR> 60 mL/min/1.73 m 2. Office blood pressure was measured by trained study personnel in a standardized way and hypertension awareness and medication were assessed during standardized interviews. Blood pressure was considered as controlled if systolic 90%. However, only 2456 (49.3%) of the hypertensive patients had controlled blood pressure. About half (51.0%) of the patients with uncontrolled blood pressure met criteria for resistant hypertension. Factors associated with better odds for controlled blood pressure in multivariate analyses included younger age, female sex, higher income, low or absent proteinuria, and use of certain classes of antihypertensive medication. We conclude that blood pressure control of CKD patients remains challenging even in the setting of nephrology specialist care, despite high rates of awareness and medication use

Single Cell Analysis Facilitates Staging of Blimp1-Dependent Primordial Germ Cells Derived from Mouse Embryonic Stem Cells

The cell intrinsic programming that regulates mammalian primordial germ cell (PGC) development in the pre-gonadal stage is challenging to investigate. To overcome this we created a transgene-free method for generating PGCs in vitro (iPGCs) from mouse embryonic stem cells (ESCs). Using labeling for SSEA1 and cKit, two cell surface molecules used previously to isolate presumptive iPGCs, we show that not all SSEA1+/cKit+ double positive cells exhibit a PGC identity. Instead, we determined that selecting for cKitbright cells within the SSEA1+ fraction significantly enriches for the putative iPGC population. Single cell analysis comparing SSEA1+/cKitbright iPGCs to ESCs and embryonic PGCs demonstrates that 97% of single iPGCs co-express PGC signature genes Blimp1, Stella, Dnd1, Prdm14 and Dazl at similar levels to e9.5–10.5 PGCs, whereas 90% of single mouse ESC do not co-express PGC signature genes. For the 10% of ESCs that co-express PGC signature genes, the levels are significantly lower than iPGCs. Microarray analysis shows that iPGCs are transcriptionally distinct from ESCs and repress gene ontology groups associated with mesoderm and heart development. At the level of chromatin, iPGCs contain 5-methyl cytosine bases in their DNA at imprinted and non-imprinted loci, and are enriched in histone H3 lysine 27 trimethylation, yet do not have detectable levels of Mvh protein, consistent with a Blimp1-positive pre-gonadal PGC identity. In order to determine whether iPGC formation is dependent upon Blimp1, we generated Blimp1 null ESCs and found that loss of Blimp1 significantly depletes SSEA1/cKitbright iPGCs. Taken together, the generation of Blimp1-positive iPGCs from ESCs constitutes a robust model for examining cell-intrinsic regulation of PGCs during the Blimp1-positive stage of development

Genetic newborn screening and digital technologies: A project protocol based on a dual approach to shorten the rare diseases diagnostic path in Europe.

Since 72% of rare diseases are genetic in origin and mostly paediatrics, genetic newborn screening represents a diagnostic "window of opportunity". Therefore, many gNBS initiatives started in different European countries. Screen4Care is a research project, which resulted of a joint effort between the European Union Commission and the European Federation of Pharmaceutical Industries and Associations. It focuses on genetic newborn screening and artificial intelligence-based tools which will be applied to a large European population of about 25.000 infants. The neonatal screening strategy will be based on targeted sequencing, while whole genome sequencing will be offered to all enrolled infants who may show early symptoms but have resulted negative at the targeted sequencing-based newborn screening. We will leverage artificial intelligence-based algorithms to identify patients using Electronic Health Records (EHR) and to build a repository "symptom checkers" for patients and healthcare providers. S4C will design an equitable, ethical, and sustainable framework for genetic newborn screening and new digital tools, corroborated by a large workout where legal, ethical, and social complexities will be addressed with the intent of making the framework highly and flexibly translatable into the diverse European health systems

Review of biochar role as additive in anaerobic digestion processes

because of the urgent need to provide renewable energy sources and efficiently manage the continuously growing amount of organic waste. Biochar (BC) is an extremely versatile material, which could be produced by carbonization of organic materials, including biomass and wastes, consistently with Circular Economy principles, and “tailor-made” for specific applications. The potential BC role as additive in the control of the many wellknown critical issues of AD processes has been increasingly explored over the past few years. However, a clear and comprehensive understanding of the connections between BC and AD is still missing. This review paper analyses and discusses significant references (review articles, research papers and international databases and reports), mostly published in the last 10 years. This review is aimed at addressing three key issues related to the better understanding of the BC role in AD processes: 1. Investigation of the influence of BC properties on AD performances and of their ability to counteract its main challenges; 2. Assessment of the optimal BC production chain (i.e. feedstock-pyrolysis-activation) to achieve the desired features; 3. Evaluation of the economic and environmental advantages connected to BC use in AD processes, compared to conventional solutions applied to address AD challenges