Impact of parental attributes on the oral health-related quality of life of Brazilian preschool children.

PURPOSE: Considering the importance of parental attitudes and their influence on children's oral health, it is important to know which characteristics may impact it. The aim of this population study was to evaluate the impact of the parental sociodemographic characteristics on the oral health-related quality of life (OHRQoL) of preschool children from southeastern Brazil. METHODS: A population-based study was conducted with a sample of 321 children aged 2 to 5 years old who underwent a calibrated clinical oral examination to assess oral health status and their parents were asked to answer the two following questionnaires: (1) sociodemographic conditions and (2) Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS). Descriptive analysis, Mann-Whitney and Kruskal-Wallis tests and adjusted Poisson regression models were used. RESULTS: Younger mothers (RP: 1.47; 95% CI: 1.11-1.95), lower household income (RP: 1.47; 95% CI: 1.11-1.96) and the presence of dental caries (RP: 2.28; 95% CI: 1.74-3.0) were more likely to have a negative impact on the OHRQoL B-ECOHIS total scores (P < 0.01). CONCLUSION: The characteristics of mothers, such as age, schooling and salary, influenced the mean B-ECOHIS total score; however, paternal attributes were not associated with a negative impact on the children's quality of life. Also, dental caries and household income influenced the children's oral health-related quality of life.

Trypanosomatids in Phlebotomine Sand Flies (Diptera: Phlebotominae) From Anthropic and Sinantropic Landscapes in a Rural Settlement in the Brazilian Amazon.

Trypanosomatids (Kinetoplastida:Trypanosomatidae) protozoa are a diverse group of obligate parasites. The genera Trypanosoma and Leishmania are the most studied because of their medical importance. This work aims to evaluate the effects of anthropization processes on the composition of the phlebotomine sand fly fauna and the natural infection by Trypanosomatids, with emphasis on Leishmania. At all 3,186 sand flies were collected, distributed in 13 genera and 52 species, being Ny. umbratilis the most abundant species. There was no difference in the diversity between canopy and soil environments. The species abundance and richness were higher in the forest environment while species diversity and evenness were highest in the forest edge. The ITS1 region was used by PCR-RFLP to identify the fragment profiles of Leishmania species, followed by genetic sequencing. Here were analyzed 100 pools of female sand flies, being six positive for DNA parasite. PCR-RFLP fragment patterns similar to Endotrypanum sp. were observed in Nyssomyia anduzei, Psychodopygus amazonensis and Lutzomyia gomezi, and those fragments similar to Leishmania (Leishmania) amazonensis were observed in Bichromomyia flaviscutellata. ITS1 sequencing confirmed the presence of Leishmania sp. in Bi. flaviscutellata, and Leishmania (Viannia) naiffi in Ny. anduzei, Psychodopygus amazonensis, and Lu. gomezi. This is the first record of Lu. gomezi and Ps. amazonensis infection by L. naiffi in the State of Amazonas. These results show the trypanosomatid infection in sandflies from different landscapes in a rural settlement, and the finding of species infected with L.(V.) naiffi suggest that they can develop a role in the transmission cycle of leishmaniasis.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs / Presença de múltiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de água doce (Aegla)

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos(≤ 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídicadas membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs

Abstract Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams ( 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Resumo Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos ( 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídica das membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs / Presença de múltiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de água doce (Aegla)

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos (≤ 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídica das membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Evaluation of eight protocols for genomic DNA extraction of Hypostomus commersoni Valenciennes, 1836 (Loricariidae: Siluriformes) / Avaliação de oito protocolos na extração de DNA genômico de Hypostomus commersoni Valenciennes, 1836 (Loricariidae: Siluriformes)

Abstract The principle and the techniques applied in DNA extraction play a pivotal role in the obtention of a purified genetic material. The present study investigates the efficiency of eight protocols in the DNA extraction of Hypostomus commersoni, an essential component of South American freshwater ichthyofauna. The quality of samples was assessed through spectrophotometry, gel electrophoresis, and PCR-RAPD markers amplification. The efficiency of DNA extraction was influenced both by the method applied and the target-tissue of choice. Higher concentrations and yield of DNA were obtained from ocular tissue, with a positive spectrum of incubation in lysis buffer for up to 36 hours after sample collection, using fresh tissues and in the presence of a high concentration of Proteinase K (20 mg.ml-1). In these conditions, samples were successfully amplified. To date, there is no record of description for the parameters analyzed in this work, neither the description of RAPD markers for the species H. commersoni.

Resumo Os princípios e as técnicas aplicadas na extração de DNA desempenham um papel crucial na obtenção de material genético purificado. O presente estudo investiga a eficiência de oito protocolos na extração de DNA de Hypostomus commersoni, um importante componente ictiofaunístico de riachos da América do Sul. A qualidade das amostras foi avaliada por espectrofotometria, eletroforese em gel e amplificação por marcadores de PCR-RAPD. A eficiência da extração de DNA foi influenciada tanto pelo método aplicado quanto pelo tecido-alvo de escolha. Maiores concentrações e rendimento de DNA foram obtidos a partir do tecido ocular, com um espectro positivo de incubação em tampão de lise por até 36 horas após a coleta da amostra, utilizando tecidos frescos e na presença de alta concentração de proteinase K (20 mg.ml-1). Nestas condições, as amostras foram amplificadas com sucesso. Até o momento, não há registro de descrição para os parâmetros analisados neste trabalho, nem a descrição de marcadores RAPD para a espécie H. commersoni.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs.

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Evaluation of eight protocols for genomic DNA extraction of Hypostomus commersoni Valenciennes, 1836 (Loricariidae: Siluriformes).

The principle and the techniques applied in DNA extraction play a pivotal role in the obtention of a purified genetic material. The present study investigates the efficiency of eight protocols in the DNA extraction of Hypostomus commersoni, an essential component of South American freshwater ichthyofauna. The quality of samples was assessed through spectrophotometry, gel electrophoresis, and PCR-RAPD markers amplification. The efficiency of DNA extraction was influenced both by the method applied and the target-tissue of choice. Higher concentrations and yield of DNA were obtained from ocular tissue, with a positive spectrum of incubation in lysis buffer for up to 36 hours after sample collection, using fresh tissues and in the presence of a high concentration of Proteinase K (20 mg.ml-1). In these conditions, samples were successfully amplified. To date, there is no record of description for the parameters analyzed in this work, neither the description of RAPD markers for the species H. commersoni.

Effects of Hyperosmolality on Hypothalamic Astrocytic Area, mRNA Expression and Glutamate Balance In Vitro.

During prolonged dehydration, body fluid homeostasis is challenged by extracellular fluid (ECF) hyperosmolality, which induce important functional changes in the hypothalamus, in parallel with other effector responses, such as the activation of the local renin-angiotensin system (RAS). Therefore, in the present study we investigated the role of sodium-driven ECF hyperosmolality on glial fibrillary acid protein (GFAP) immunoreactivity and protein expression, membrane capacitance, mRNA expression of RAS components and glutamate balance in cultured hypothalamic astrocytes. Our data show that hypothalamic astrocytes respond to increased hyperosmolality with a similar decrease in GFAP expression and membrane capacitance, indicative of reduced cellular area. Hyperosmolality also downregulates the transcript levels of angiotensinogen and both angiotensin-converting enzymes, whereas upregulates type 1a angiotensin II receptor mRNA. Incubation with hypertonic solution also decreases the immunoreactivity to the membrane glutamate/aspartate transporter (GLAST) as well as tritiated-aspartate uptake by astrocytes. This latter effect is completely restored to basal levels when astrocytes previously exposed to hypertonicity are incubated under isotonic conditions. Together with a direct effect on two important local signaling systems (glutamate and RAS), these synaptic rearrangements driven by astrocytes may accomplish for a coordinated increase in the excitatory drive onto the hypothalamic neurosecretory system, ultimately culminating with increased AVP release in response to hyperosmolality.

Comparison of Two Models of Frailty for the Prediction of Mortality in Brazilian Community-Dwelling Older Adults: The FIBRA Study.

BACKGROUND: Prevalence of frailty is significant in Latin America. However, no previous study evaluated mortality prediction using the two most used frailty models in Brazil. OBJECTIVES: The aim of the present study was to compare the frailty phenotype and the frailty index with regard to accuracy in the prediction of mortality among community-dwelling older adults. METHODS: A cohort study was conducted involving 674 older adults. Thirty-five variables (signs, symptoms, chronic diseases and disabilities) were used for the construction of the frailty index (FI). The frailty phenotype index (FPI) was defined based on the criteria proposed in the Cardiovascular Health Study. Periodic verifications were performed in the databank of the Mortality Information System. Cox regression was used to estimate the relative risk (RR) of mortality and Kaplan-Meier survival curves were used in the analysis. RESULTS: The prevalence of frailty was greater based using the FI (16.3%) compared to the FPI (5.34%). Older adults classified as frail by the FPI had a greater risk of death (RR: 10.03; 95% CI: 4.43-22.74) that those classified as frail by the FI (RR: 0.87; 95% CI: 0.25-3.00). The lowest survival rate was found in the group of older adults classified as frail based on the FPI and classified as pre-frail and robust based on the FI. CONCLUSION: The FPI demonstrated greater accuracy in predicting the risk of mortality among Brazilian older adults than the FI. The validation of frailty measures is fundamental to the identification of older adults who are more vulnerable to adverse health events.

Adiponectin serum levels in female dogs bearing mammary gland carcinomas / Níveis séricos de adiponectina em cadelas portadoras de carcinomas mamários

Em seres humanos, a adiponectinemia está associada à obesidade e ao risco aumentado a uma ampla variedade de cânceres. Embora o papel dessa adipocina esteja bem documentado na patogênese do câncer em humanos, tal associação permanece a ser determinada em cães. Nesses animais, a relação da adiponectina com a carcinogênese parece ser ainda meramente especulativa. Nesse contexto, buscou-se nesta investigação comparar os níveis séricos de adiponectina em fêmeas hígidas e em portadoras de carcinomas mamários com diagnóstico histopatológico de carcinoma mamário tubular simples estágio 4, com comprometimento de linfonodos, porém sem metástases a distância detectadas. Foi observado que as cadelas diagnosticadas com carcinoma mamário tiveram níveis séricos de adiponectina significativamente menores (média de 3,72±1,54µg/mL, P<0,05) em relação às fêmeas consideradas hígidas (média de 5,61±2,18µgl/mL), sugerindo associação entre câncer e hipoadiponectinemia similar à encontrada em humanos. Em relação à idade e ao peso corporal dos animais do estudo, não foi encontrada diferença significativa entre os grupos. Os resultados encontrados nas cadelas portadoras de carcinoma mamário do presente estudo corroboram a associação já descrita em humanos entre ocorrência de carcinogênese e baixos níveis de adiponectina.(AU)

Effects of peanut drying and blanching on Salmonella spp.

In order to evaluate the behavior of Salmonella during peanut drying and blanching, a study was conducted with Runner type peanuts. Samples of raw in-shell or unblanched peanuts were inoculated by spraying with a pool of five Salmonella serotypes isolated from the peanut supply chain (Miami, Muenster, Yoruba, Javiana and Glostrup). The in-shell peanuts were submitted to drying at 35 and 40 °C up to 18 h. After this time, the Salmonella counts went down ca. 2.0 log MPN/g at 35 and 40 °C. According to the Weibull model the time needed to achieve Salmonella 3-log reduction (T3d) and 5-log reduction (T5d) on the in-shell peanuts would be ca. 49 and 117 h at 35 °C and 35 and 79 h at 40 °C, respectively. The results showed that there was no statistical difference (p > .05) between either of the temperatures employed in the process. The blanching process was performed in two steps: pre-roasting (step 1) and skin removal (step 2). Reduction of up to 2.1 log MPN/g was observed after blanching at 100 °C/15 min plus 15 s of air impact. The skin removal process did not result in recontamination of the final sample. The Weibull model predicted 3- and 5-log reductions of Salmonella in 37.0 and 68.9 min for blanching at 95 °C, and in 39.1 and 114.9 min at 100 °C. The results demonstrated that drying and blanching processes did not generate large reductions of Salmonella in the peanut samples. Thus, the product resulting from these steps may be a possible source of cross-contamination for the processing plant and the final product.

Anti-EGFR-resistant clones decay exponentially after progression: implications for anti-EGFR re-challenge.

BACKGROUND: Colorectal cancer (CRC) has been shown to acquire RAS and EGFR ectodomain mutations as mechanisms of resistance to epidermal growth factor receptor (EGFR) inhibition (anti-EGFR). After anti-EGFR withdrawal, RAS and EGFR mutant clones lack a growth advantage relative to other clones and decay; however, the kinetics of decay remain unclear. We sought to determine the kinetics of acquired RAS/EGFR mutations after discontinuation of anti-EGFR therapy. PATIENTS AND METHODS: We present the post-progression circulating tumor DNA (ctDNA) profiles of 135 patients with RAS/BRAF wild-type metastatic CRC treated with anti-EGFR who acquired RAS and/or EGFR mutations during therapy. Our validation cohort consisted of an external dataset of 73 patients with a ctDNA profile suggestive of prior anti-EGFR exposure and serial sampling. A separate retrospective cohort of 80 patients was used to evaluate overall response rate and progression free survival during re-challenge therapies. RESULTS: Our analysis showed that RAS and EGFR relative mutant allele frequency decays exponentially (r2=0.93 for RAS; r2=0.94 for EGFR) with a cumulative half-life of 4.4 months. We validated our findings using an external dataset of 73 patients with a ctDNA profile suggestive of prior anti-EGFR exposure and serial sampling, confirming exponential decay with an estimated half-life of 4.3 months. A separate retrospective cohort of 80 patients showed that patients had a higher overall response rate during re-challenge therapies after increasing time intervals, as predicted by our model. CONCLUSION: These results provide scientific support for anti-EGFR re-challenge and guide the optimal timing of re-challenge initiation.

Preactivation of neutrophils and systemic oxidative stress in dogs with hyperleptinemia.

High occurrence of obesity currently constitutes the main nutritional disease of the canine species. There is evidence that leptin increases during obesity in dogs. Hyperleptinemia is associated with increased neutrophil oxidative metabolism in obese humans and contributes to oxidative stress. However, in obese dogs, the probable relationship between this condition and the activation of the oxidative metabolism of neutrophils has yet to be established. Thus, we investigated the hypothesis that neutrophil activation and systemic oxidative stress occur in dogs with hyperleptinemia. A control group of 24 healthy dogs with a body condition score (BCS) of 4-5, an overweight group of 25 dogs with a BCS of 6-7, and 27 obese dogs with a BCS of 8-9, were composed. Two subgroups were formed composed of dogs with and without hyperleptinemia, grouped according to the 95% confidence interval obtained for plasma leptin values of the control group. Changes in obesity markers (body condition score, adiponectin and plasma leptin) and plasma oxidative stress (lipid peroxidation, total antioxidant and oxidant capacities and oxidative stress index) were measured in all the dogs selected. Neutrophil oxidative metabolism was evaluated in flow cytometry by superoxide production with the probe hydroethidine and by hydrogen peroxide production with the probe 2',7'-dichlorofluorescein diacetate, with or without phorbol myristate acetate (PMA) stimulation. Apoptosis and neutrophil viability were quantified in a capillary flow cytometer using Annexin VPE, with or without camptothecin apoptosis inducing effect. Obese dogs presented higher systemic oxidative stress, hyperleptinemia and preactivated neutrophils with accelerated apoptosis. Dogs with hyperleptinemia and obese dogs presented higher neutrophil superoxide production under PMA stimulation and the presence of systemic oxidative stress compared with control. To our knowledge, this is probably the first evidence that preactivation of the oxidative metabolism of circulating neutrophils occurs in dogs with hyperleptinemia, a condition that can induce systemic oxidative stress in the canine species.

Characterization of oxidative stress biomarkers in a freshwater anomuran crab / Caracterização de biomarcadores de estresse oxidativo em caranguejos anomuros de água doce

Abstract In general, environmental responses at level of populations or communities are preceded by alterations at lower biological levels which can be efficiently detected by the analysis of biomarkers. We analyzed the oxidative biomarkers TBARS and Catalase in Aegla singularis, a freshwater crustacean highly sensitive to environmental changes. The objective was to address if are differences in these biomarkers related to the gender as well if they are influenced by seasonal or water physicochemical variables. The results showed differences in biomarkers profile related to the gender. In female crabs were not sensitive to seasonal variations throughout the study period. However, in males the biomarkers evaluated were higher in the winter as compared to remaining seasons and showed tendency of negative correlation with water temperature and pH. This study highlights that gender, seasonal variations and physicochemical variables can influence oxidative stress biomarkers in A. singularis. Female crabs probably are better suited as a model for biomarker application in environmental studies, because their insensibility to seasonal variations can facilitate the observations of responses related specifically to environmental disturbances.

Resumo Em geral, as respostas ambientais ao nível de populações ou comunidades são precedidas pelas alterações nos níveis biológicos inferiores que podem ser eficientemente detectados pela análise de biomarcadores. Neste trabalho, foram analisados os biomarcadores oxidativos TBARS e Catalase em Aegla singularis, um crustáceo de água doce altamente sensível às mudanças ambientais. O objetivo foi investigar se há diferenças nestes biomarcadores relacionados com o gênero, bem como se eles são influenciados por parâmetros sazonais ou físico-químicos. Os resultados mostraram diferenças no perfil de biomarcadores relacionados com o gênero. Caranguejos fêmeas não foram sensíveis a variações sazonais ao longo do período de estudo. Nos machos, os biomarcadores avaliadas apresentaram níveis mais altos no inverno, em comparação com as demais estações e mostraram uma tendência de correlação negativa com a temperatura e pH da água. Este estudo destaca que o sexo, variações sazonais e variáveis físico-químicas podem influenciar os biomarcadores de estresse oxidativo em A. singularis. As fêmeas de A. singularis provavelmente são mais adequadas como um modelo para aplicação destes biomarcadores em estudos ambientais, uma vez que sua insensibilidade às variações sazonais podem facilitar as observações das respostas relacionadas especificamente com perturbações ambientais.

Characterization of oxidative stress biomarkers in a freshwater anomuran crab.

In general, environmental responses at level of populations or communities are preceded by alterations at lower biological levels which can be efficiently detected by the analysis of biomarkers. We analyzed the oxidative biomarkers TBARS and Catalase in Aegla singularis, a freshwater crustacean highly sensitive to environmental changes. The objective was to address if are differences in these biomarkers related to the gender as well if they are influenced by seasonal or water physicochemical variables. The results showed differences in biomarkers profile related to the gender. In female crabs were not sensitive to seasonal variations throughout the study period. However, in males the biomarkers evaluated were higher in the winter as compared to remaining seasons and showed tendency of negative correlation with water temperature and pH. This study highlights that gender, seasonal variations and physicochemical variables can influence oxidative stress biomarkers in A. singularis. Female crabs probably are better suited as a model for biomarker application in environmental studies, because their insensibility to seasonal variations can facilitate the observations of responses related specifically to environmental disturbances.

M2 macrophages coexist with a Th1-driven profile in periapical cysts.

AIM: To evaluate the participation of both Th1 and Th2 responses in periapical cysts by assessing the presence of M2 macrophages, as well as acute IL-1 ß, TNF-α and IL-6 cytokines. METHODOLOGY: Twenty-four cases of periapical cysts were selected. Immuno-expressions of IL-1 ß, IL-6, TNF-α and CD163 were analysed in the cystic capsules in both superficial and deeper regions. Data were analysed with paired Wilcoxon test and Spearman correlation coefficient (P ≤ 0.05). RESULTS: There was a higher expression of IL-1ß, IL-6, TNF-α and M2 macrophages in the superficial region (P < 0.001) of cystic capsules. All acute cytokines had significant positive correlations amongst them regardless of the cystic capsule region. Regarding CD163, positive correlations occurred only with TNF-α (P = 0.007; r = 0.537) and IL-6 (P = 0.018; r = 0.478) in the superficial regions of the cystic capsule. CONCLUSIONS: M2 macrophages participated actively in the inflammatory response of periapical cysts and correlated with the expression of certain acute Th1-related cytokines. This illustrates the coexistence of an acute and chronic Th2-driven immune response in these lesions. Although M2 macrophages favour the healing process, their presence is not sufficient for periapical cyst regression, once an acute active response has occurred due to an infectious stimuli.

Frontal alignment in total knee arthroplasty. Comparative study between radiographic measurement and surgical navigation. / Alineación frontal en la artroplastia total de rodilla. Estudio comparativo entre medición radiográfica y con navegación quirúrgica.

OBJECTIVE: To establish the concordance between angulation of the femoro-tibial mechanical axis measured with x-rays and surgical navigation in both the pre and postoperative period after a total knee arthroplasty (TKA) was implanted. MATERIAL AND METHOD: Pre and postoperative measurements were analyzed in 88 TKA of the same model and all performed with the same surgical navigation system. The mechanical frontal angle (MFA) and femoro-tibial anatomic angle were measured before and after the TKA. The angulation was digitally measured with a teleradiography. In the navigation, the femoro-tibial angle at rest, forced varus and valgus were registered and the average of these three measurements was calculated. RESULTS: The mean preoperative MFA measured on the radiograph was 4.55°. The mean of the same angle measured on the postoperative radiograph was 1.72°, (p=0.05). The mean of the MFA measured with navigation before TKA was 3.12° and after the implant with navigation was 0.53 (P=.013). The concordance coefficient between the MFA in teleradiography and in navigation was 0.869 (P<.001) preoperatively and 0.709 postoperatively (P=.017). CONCLUSIONS: We found a strong concordance between radiographic and surgical navigation measurements of the MFA. This may imply that teleradiography is not necessary when using surgical navigation in TKA.