Innovative hurdle system towards Listeria monocytogenes inactivation in a fermented meat sausage model - high pressure processing assisted by bacteriophage P100 and bacteriocinogenic Pediococcus acidilactici

Consumers' quest for healthier, locally produced foods, renders the demand for these products increasingly prominent. The purpose of the present work was to evaluate the impact of a non-thermal multi-hurdle approach, which associated mild high hydrostatic pressure (HHP, 300 MPa), the bacteriophage Listex™ P100, and the pediocin PA-1 producing Pediococcus acidilactici HA 6111-2, as a novel minimal processing towards Listeria monocytogenes eradication in Alheira (a traditional fermented meat sausage from Northern Portugal). The combination of the three hurdles achieved the USDA-FSIS 5 log reduction (in accordance with the standard guidelines for ready-to-eat foods), being the only treatment to elicit the absence of L. monocytogenes immediately following processing (p  0.05) in the pH values were observed, and the semi-quantification of the in situ biosynthesized pediocin PA-1 was documented for the first time in a fermented meat sausage model.publishe

Effects of consumption of galactooligosaccharides obtained through whey enzymatically modified on the faecal flora and nutritional parameters of hamsters

The aim of this research was to evaluate the influence of wheyenzymatically modified rich in galactooligosaccharides in thenutritional characteristics and effects in the microflora of cecumcontents by the study with Golden Syrian hamsters (Mesocricetusauratus) for 28 days (controlled conditions). Three isoproteic dietswere prepared (20% w/w): C (casein), W (whey) and G (wheymodified). The groups studied differed positively from the C regardingfeed and protein efficiency ratio. The relationships (w/w) oforgan/body were found proportional in all diets. The counts ofprobiotics from the cecum contents the groups showed no difference.The pHs of studied groups were lower than C, this acidity can atimpairs the ability of pathogens to grow in the intestine. Resultssuggest that using whey enzymatically modified rich ingalactooligosaccharides could replace the standard diet withnutritional efficiency and possible inhibit the microorganismspathogenic without induce damage in health.Fil: Dos Santos Da Fonseca, Renata Aline. Universidade Federal Do Rio Grande; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Rodrigues Machado, Adriana. Universidade Federal Do Rio Grande; BrasilFil: Muniz Moreira, Lidiane. Universidade Federal Do Rio Grande; BrasilFil: Rodrigues, Rosane S.. Universidade Federal de Pelotas; BrasilFil: Machado, Mirian. Universidade Federal de Pelotas; BrasilFil: Souza Soares, Leonor A.. Universidade Federal Do Rio Grande; BrasilFil: Burkert, Carlos André V.. Universidade Federal Do Rio Grande; BrasilFil: Burkert, Janaína Fernandes de Medeiros. Universidade Federal Do Rio Grande; Brasi

Convolutamydine A and synthetic analogues have antinociceptive properties in mice

AbstractConvolutamydine A, an oxindole that originated from a marine bryozoan, has several biological effects. In this study, we aimed to investigate the antinociceptive effects of convolutamydine A and two new synthetic analogues.Convolutamydine A and the two analogues were given orally to assess their ability to induce antinociceptive effects. Formalin-induced licking response, acetic acid-induced contortions, and hot plate models were used to characterize the effects of convolutamydine A and its analogues.Convolutamydine A (4,6-bromo-3-(2-oxopropyl)-3-hydroxy-2-oxindole), compound 1 (3-(2-oxopropyl)-3-hydroxy-2-oxindole), and compound 2 (5-bromo-3-(2-oxopropyl)-3-hydroxy-2-oxindole) caused peripheral antinociceptive and anti-inflammatory effects in the acetic acid-induced contortions and the formalin-induced licking models. Supraspinal effects were also observed in the hot plate model and were similar to those obtained with morphine. The peripheral effects were not mediated by the cholinergic or opioid systems. The antinociceptive effects of convolutamydine A seem to be mediated by all three systems (cholinergic, opioid, and nitric oxide systems), and the mechanism of action of compounds 1 and 2 involved cholinergic and nitric oxide-mediated mechanisms. Convolutamydine A and its analogues (compounds 1 and 2) showed good antinociceptive ability after systemic administration in acute pain models. The antinociceptive action mediated by cholinergic, opioid, and nitric oxide systems could explain why convolutamydine A, compound 1, and compound 2 retained their antinociceptive effects. The doses used were similar to the doses of morphine and were much lower than that of acetylsalicylic acid, the classical analgesic and anti-inflammatory drug.In conclusion, convolutamydine A and the two analogues demonstrated antinociceptive effects comparable to morphine's effects

Next-generation Sequencing-based genomic profiling: Fostering innovation in cancer care?

OBJECTIVES: With the development of next-generation sequencing (NGS) technologies, DNA sequencing has been increasingly utilized in clinical practice. Our goal was to investigate the impact of genomic evaluation on treatment decisions for heavily pretreated patients with metastatic cancer. METHODS: We analyzed metastatic cancer patients from a single institution whose cancers had progressed after all available standard-of-care therapies and whose tumors underwent next-generation sequencing analysis. We determined the percentage of patients who received any therapy directed by the test, and its efficacy. RESULTS: From July 2013 to December 2015, 185 consecutive patients were tested using a commercially available next-generation sequencing-based test, and 157 patients were eligible. Sixty-six patients (42.0%) were female, and 91 (58.0%) were male. The mean age at diagnosis was 52.2 years, and the mean number of pre-test lines of systemic treatment was 2.7. One hundred and seventy-seven patients (95.6%) had at least one identified gene alteration. Twenty-four patients (15.2%) underwent systemic treatment directed by the test result. Of these, one patient had a complete response, four (16.7%) had partial responses, two (8.3%) had stable disease, and 17 (70.8%) had disease progression as the best result. The median progression-free survival time with matched therapy was 1.6 months, and the median overall survival was 10 months. CONCLUSION: We identified a high prevalence of gene alterations using an next-generation sequencing test. Although some benefit was associated with the matched therapy, most of the patients had disease progression as the best response, indicating the limited biological potential and unclear clinical relevance of this practice

Atherosclerosis and Bone Loss in Humans–Results From Deceased Donors and From Patients Submitted to Carotid Endarterectomy

Funding: We wish to thank all the collaborators (administrative staff, nurses, etc.) of the surgery block, as well as the doctors of the vascular surgery and transplantation departments of the Hospital of Santa Maria for the availability and assistance in the collection of the samples. We also thank Sociedade Portuguesa de Reumatologia for funding with two fellowships: Fundo de Apoio à Investigação 2014 and SPR/MSD 2015. DC-F received funding from a PhD grant from Fundação para a Ciência e a Tecnologia (SFRH/BD/80940/2011).Background and Aims: Atherosclerosis and osteoporosis share common risk factors, as well as inflammatory mechanisms. Our aim was to understand how atherosclerotic lesions are related with disturbances in bone. Methods: Gene expression of pro-inflammatory and bone metabolism related proteins (IL-1β, IL-6, IL-17A, TNF, RANKL, OPG, COL1, CTSK, OCL, TRAP, CBFA1, DKK1, SOST, ADIPOQ, and ADIPOR1) were analyzed in arteries and bones from 45 deceased donors and adipose tissue was used as control. Additionally, in 139 patients with advanced atherosclerosis submitted to carotid endarterectomy we compared calcium content (Alizarin red) and plaque inflammatory scores (CD3+, CD68+, and adiponectin) of patients with normal bone mineral density (BMD) with those with low BMD and explored the associations between gene expression in atherosclerotic plaques and BMD. Serum levels of pro-inflammatory and bone related proteins were measured both in donors and patients. Associations were investigated by the Pearson or Spearman correlation tests, and multivariate regression analyzes were performed when justified. Results: Gene expression of bone remodeling and pro-inflammatory proteins correlated positively in bone and aorta, independently of age and sex of donors, but not in adipose tissue. The expression of bone formation genes was significantly higher in atheroma plaques from endarterectomized patients with normal vs. low BMD as well as inflammatory CD68+ scores, regardless of patients' age and sex, but not of body mass index. No relationship was observed between serum levels and gene expression levels of pro-inflammatory or bone remodeling proteins. Conclusions: Our results suggest that the relationship between bones and vessels in the context of atherosclerotic disease and osteoporosis may rely on the intrinsic connection between the tissues involved, independently of disease stage. Serum measurements of pro-inflammatory and bone-remodeling proteins do not accurately translate tissue pathologic processes.publishersversionpublishe

Thoracic cirtometry in children with Duchenne muscular dystrophy - expansion of the method

BACKGROUND: Thoracic cirtometry is a simple and accessible technique to evaluate chest mobility during forced breathing. However, it does not allow for the assessment of compensatory movements commonly used by people with chronic diseases, such Duchenne muscular dystrophy (DMD). DMD is a condition characterized by progressive and irreversible degeneration of the musculoskeletal system. OBJECTIVES: To expand the method of thoracic cirtometry to allow for the assessment of compensatory movements; to analyze the reliability of the tool; and to describe thoracic mobility of children with DMD during deep breathing. METHOD: Sixty boys, 30 with DMD (10.1±0.5 years) and 30 healthy controls (9.5±0.6 years) participated in the study. The expanded thoracic cirtometry was organized in two phases: 1. the body could move freely, allowing the assessment of compensatory movements (free thoracic cirtometry) and 2. the body without compensatory movements, allowing for the direct study of the movements of the chest (guided thoracic cirtometry). This method includes videotaping and systematic observation of body movements using descriptive and numeric data. We investigated reliability of these measures in both groups. RESULTS: Measures of axial and the xiphoid thoracic cirtometry (both free and guided) showed excellent reliability. All measures were significantly different between groups. In DMD boys, free thoracic cirtometry presented a greater value of chest expansion when compared with the guided measures, which probably occurred due to compensatory movements. The most commons were movements of the head, shoulder and torso. CONCLUSIONS: The expanded thoracic cirtometry method showed excellent reliability and achieved the objectives of determining measures of chest mobility and compensatory movements during deep breath. We suggested its use in the respiratory evaluation of children with DMD

Non-peptidic Cruzain Inhibitors with Trypanocidal Activity Discovered by Virtual Screening and in Vitro Assay

A multi-step cascade strategy using integrated ligand-and target-based virtual screening methods was developed to select a small number of compounds from the ZINC database to be evaluated for trypanocidal activity. Winnowing the database to 23 selected compounds, 12 non-covalent binding cruzain inhibitors with affinity values (K-i) in the low micromolar range (3-60 mu M) acting through a competitive inhibition mechanism were identified. This mechanism has been confirmed by determining the binding mode of the cruzain inhibitor Nequimed176 through X-ray crystallographic studies. Cruzain, a validated therapeutic target for new chemotherapy for Chagas disease, also shares high similarity with the mammalian homolog cathepsin L. Because increased activity of cathepsin L is related to invasive properties and has been linked to metastatic cancer cells, cruzain inhibitors from the same library were assayed against it. Affinity values were in a similar range (4-80 mu M), yielding poor selectivity towards cruzain but raising the possibility of investigating such inhibitors for their effect on cell proliferation. in order to select the most promising enzyme inhibitors retaining trypanocidal activity for structure-activity relationship (SAR) studies, the most potent cruzain inhibitors were assayed against T. cruzi-infected cells. Two compounds were found to have trypanocidal activity. Using compound Nequimed42 as precursor, an SAR was established in which the 2-acetamidothiophene-3-carboxamide group was identified as essential for enzyme and parasite inhibition activities. the IC50 value for compound Nequimed42 acting against the trypomastigote form of the Tulahuen lacZ strain was found to be 10.6 +/- 0.1 mu M, tenfold lower than that obtained for benznidazole, which was taken as positive control. in addition, by employing the strategy of molecular simplification, a smaller compound derived from Nequimed42 with a ligand efficiency (LE) of 0.33 kcal mol(-1) atom(-1) (compound Nequimed176) is highlighted as a novel non-peptidic, non-covalent cruzain inhibitor as a trypanocidal agent candidate for optimization.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Fed Sao Carlos, Dept Quim, BR-13560 Sao Carlos, SP, BrazilUniv São Paulo, Inst Quim Sao Carlos, Grp Quim Med IQSC USP, Sao Carlos, SP, BrazilUniv Calif San Francisco, Dept Pathol, Ctr Discovery & Innovat Parasit Dis, San Francisco, CA 94140 USAUniv São Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, BR-14049 Ribeirao Preto, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, São Paulo, BrazilFAPESP: 2011/01893-3,CNPq: 301614/2010-5CAPES: 5985/11-0Web of Scienc

MEDICINES WASTE POLICIES AND THE POPULATION KNOWLEDGE IN BRAZIL

Objective: To investigate the degree of knowledge of the patients enrolled in the Unified Health System of the Medicine School of ABC (Faculdade de Medicina do ABC) regarding the proper use and disposal of medicaments. Methods: Participants were recruited for convenience, during their medical appointments at the clinic of the Medical School of ABC (Santo André, Sao Paulo, Brazil) in the period from 04 August to 30 September 2014. Data collection was conducted through a self-administered poll designed specifically for the purpose of this study, which consisted of 25 questions multiple choice about socioeconomic issues and the subject disposal of drugs, consumption and environmental pollution. Results: We selected the 140 patients’ polls. Most of them is of white ethnicity (58%) and female (58%). Level of education: 31% have completed secondary education (31%) or incomplete graduation (19%). Most of the participants (76%) buy drugs without a prescription, and most families (76%) seek understanding by reading the labels. 71.43% reported knowing that incorrect disposal of drugs could contaminate the environment, but 78% reported never having seen or received information about these. After using, 22.15% maintains the medication at home for future use, 55% of subjects reported improper disposal sites and 13% are delivered in health care institutions. Conclusion: Our study has showed that most participants inappropriately use and dispose of drugs, even though they know they can contaminate the environment

CIANOBACTÉRIAS E CIANOTOXINAS EM RESERVATÓRIOS DOESTADO DORIOGRANDEDONORTEE O POTENCIAL CONTROLE DAS FLORAÇÕES PELA TILÁPIA DO NILO (OREOCHROMIS NILOTICUS)

Cyanobacteria blooms in reservoirs result in loss of water quality and negative effects to human health. To reduce these impacts the monitoring of the cyanobacteria and cyanotoxin occurrence as well as the application of measures to counteract the cyanobacteria blooms are made necessary. The manipulation of filter-feeding omnivorous fish stock, as Nile tilapia, has been proposed as an strategy for cyanobacteria bloom control. The present work aimed to evaluate: I) the presence of cyanobacteria (composition, density and biovolume by sedimentation technique) and cianotoxins (mouse bioassay) in five reservoirs located at the semi-arid region of Rio Grande do Norte state, in five sampling campaigns between September 2002 and March 2004; II) the per capita consumption rates of filamentous cyanobacteria by Nile tilapia through laboratory experiments using a natural population of cyanobacteria (experiment I) and a culture of Cylindrospermopsis raciborskii (experiment II), after the juveniles tilapias being exposed to a gradient of cyanobacteria biomass. The phytoplankton of the investigated reservoirs were dominated by cyanobacteria, including various toxigenic species (C. raciborskii, Microcystis spp., Aphanizomenon e Anabaena circinalis). Cianotoxins were present in three out of five reservoirs. The consumption rate of cyanobacteria (?g chlorophyll-a.fish-1.day-1) by the tilapia was 0,29 in the experiment I and 0,5 in the experiment II. The human populations that use the studied reservoirs as drinking water supply are being potentially exposed to the negative effects of the cyanobacteria. The present research suggests that the stock of Nile tilapia to control cyanobacteria blooms is viable. Nevertheless, factors such as the ichthyo-eutrophication and the accumulation of cyanotoxins in the fish biomass should be taken into account before implementing a biomanipulation program.Florações de cianobactérias em reservatórios resultam na perda da qualidade da água e em efeitos negativos para a saúde humana. Faz-se, portanto, necessário o monitoramento da ocorrência de cianobactérias e cianotoxinas, bem como a aplicação de medidas de controle das florações. Uma estratégia viável para esse controle é a manipulação dos estoques de peixes onívoros filtradores, como a tilápia do Nilo. Neste trabalho foram avaliados: i) a presença de cianobactérias (composição, densidade e biovolume, pelo método da sedimentação) e cianotoxinas (bioensaios com camundongos) em cinco reservatórios do semi-árido do estado do Rio Grande do Norte, em cinco campanhas de coletas, entre setembro de 2002 e março de 2004; e ii) as taxas de consumo per capita de cianobactérias filamentosas pela tilápia, através de dois experimentos em laboratório, utilizando populações naturais de cianobactérias (experimento I) e uma cultura de Cylindrospermopsis raciborskii (experimento II), após a exposição de indivíduos jovens de tilápia a um gradiente de biomassa de cianobactérias. O fitoplâncton nos reservatórios estudados apresentou dominância de cianobactérias, incluindo várias espécies toxigênicas (C. raciborskii, Microcystis spp., Aphanizomenon e Anabaena circinalis). Cianotoxinas foram evidenciadas em três dos cinco reservatórios. O consumo de cianobactérias (?g clorofila-a.peixe-1.dia-1) foi de 0,29 no experimento Ie de 0,5 no experimento II. Populações abastecidas pelos reservatórios investigados estão potencialmente expostas aos efeitos negativos das cianotoxinas. Esta pesquisa sugere que é viável a estocagem da tilápia do Nilo no controle de florações de cianobactérias. Entretanto, fatores como a ictioeutrofização e o acúmulo de cianotoxinas na biomassa dos peixes devem ser levados em consideração antes que um programa de biomanipulação possa ser implementado