Light-curing of calcium hydroxide-based liners: pH analysis and calcium ion release / Fotoativação de forradores a base de hidróxido de cálcio: análise de pH e liberação de íons cálcio

Objective: Compare the pH values and calcium ion release of calcium hydroxide-based liner materials before and after light-curing. Material and Methods: The materials evaluated were: hydrox-cal white (HW), hydrox-cal dentin (HD), Biocal (BC) and UltraBlend Plus (UB). 120 samples of the liner materials were inserted into a PVC tube (n=15). The samples from HW+A, HD+A, BC+A and UB+A were subjected to photoactivation. The other groups HW+N, HD+N, BC+N and UB+N were only inserted in a glass tube with deionized water. The pH was measured 24 hours and 14 days after the inclusion of the samples with the aid of a pH meter. The calcium release was analyzed with the aid of an atomic absorption spectophotometer at 24h and 14 days. The results were submitted to the Shapiro-Wilk test, followed by ANOVA and Tukey test (p=0.05). Results: In 24h, the groups that were not light cured showed the highest pH values (p<0.05). In 14 days, BC+N and BC+A demonstrated the lowest pH values. The groups that were not light cured also showed higher calcium release values in 24h and 14 days (p<0.05). Conclusion: Photoactivation of calcium hydroxide-based liner materials negatively interferes with calcium ion release, as well as with pH.(AU)

Objetivo: Comparar os valores de pH e liberação de íons cálcio de materiais forradores à base de hidróxido de cálcio antes e depois da fotopolimerização. Material e métodos: Os materiais avaliados foram: Hidrox-cal branco (HW), Hidrox-cal dentina (HD), Biocal (BC) e UltraBlend Plus (UB). 120 amostras dos materiais de revestimento foram inseridas em um tubo de PVC (n=15). As amostras de HW +A, HD+A, BC+A e UB+A foram submetidas à fotoativação. Os demais grupos HW +N, HD+N, BC+N e UB+N foram inseridos apenas em um tubo de vidro com água deionizada. O pH foi medido 24 horas e 14 dias após a inclusão das amostras com o auxílio de um medidor de pH. A liberação de cálcio foi analisada com o auxílio de um espectrofotômetro de absorção atômica em 24h e 14 dias. Os resultados foram submetidos ao teste de Shapiro-Wilk, seguido de ANOVA e teste de Tukey (p=0,05). Resultados: Em 24h, os grupos não fotopolimerizados apresentaram os maiores valores de pH (p<0,05). Em 14 dias, BC+N e BC+A apresentaram os menores valores de pH. Os grupos não fotopolimerizados também apresentaram maiores valores de liberação de cálcio em 24h e 14 dias (p<0,05). Conclusão: A fotoativação de materiais de revestimento à base de hidróxido de cálcio interfere negativamente na liberação de íons cálcio e no pH (AU)

Randomized trial with soaps - what happens to a newborn baby's skin?

Abstract Objective: Evaluate the effectiveness of a children's soap with physiological pH in maintaining cutaneous pH and moisture of the newborn (NB)'s skin after the first bath. Methods: Randomized, controlled and double-blind clinical trial in a rooming-in of a tertiary maternity hospital in southern Brazil with 204 newborns > 34 gestational weeks. Gestational and obstetric history was evaluated, and newborns were randomized into two groups according to the product applied in the bath: the control group (CG), which used common liquid soap with pH 7.0 and experimental group (EG), which used children's liquid soap with pH 5.8. Evaluation was made immediately before and after bath with skin pH measurement, corneometry and clinical parameters (erythema, scaling and moisture), on the forehead, abdomen and thigh. Results: There was no difference between groups regarding gestational, obstetric and family history (p > 0.05). In CG, skin pH increased in the abdomen and thigh (p < 0.05). In EG there was an improvement in clinical parameters after bathing with: increased moisture, less erythema and less scaling (p < 0.05). On the forehead, there was a significant increase in pH after bathing (p < 0.001) similar in both groups, although no use of soap. There was no difference in corneometry between groups after bathing. Conclusions: Children's liquid soap with physiological pH maintained the acidic skin pH and moisture of the newborn's skin after the first bath, which reinforces the importance of using products with physiological pH in the hygiene of newborns. Registration number RBR-9ky84vd.

LIX1-like protein promotes liver cancer progression

Limb and CNS expressed 1 like (LIX1L) is over-expressed in several types of tumors. However, the function of LIX1L in glucose metabolism and hepatocellular carcinoma (HCC) progression remains elusive. Here we report that LIX1L is over-expressed in human HCC tissues, which predicts unfavorable prognosis. LIX1L deficiency

Anaerobic co-digestion of swine manure with sweet potato or cassava in different C/N ratios / Co-digestão anaeróbia dos dejetos de suínos com batata doce ou mandioca em diferentes relações C/N

ABSTRACT: Anaerobic co-digestion (AcoD) of waste is a method of increasing methane (CH4) yield and improving biofertilizer quality. This study aimed to evaluate the best AcoD conditions for swine manure (SM) with sweet potato (SP) or cassava (C) in different amounts in semi-continuous biodigesters. Initially, using batch biodigesters, an AcoD test of the SM with SP or C was performed, adopting carbon/nitrogen (C/N) ratios of 10/1, 13/1, 17/1, and 22/1. Based on the results, a C/N ratio of 10/1 was chosen, which was the proportion that resulted in the highest reduction of volatile solids (VS) and specific biogas production. From these results, the experiment was carried out in semi-continuous biodigesters, consisting of three treatments (control (SM), SP 10/1, and C 10/1) with five replicates and a hydraulic retention time (HRT) of 30 days. Total solid (TS) and volatile solid (VS) reductions, biogas and CH4 yields, alkalinity, and volatile acidity were measured. The control treatment differed from the others and resulted in decreased biogas yield (752 LN.kgVSadded -1), CH4 (449 LN.kgVSadded -1), and CH4 content (59.7%). The AcoD treatments (SP and C) did not differ significantly for biogas yield (respectively, 901 and 883 LN.kgVSadded -1) and CH4 (respectively, 590 and 547 LN.kgVSadded -1); however, they differed in CH4 content (65.5% and 61.9% respectively). The treatments showed general reduction averages of 76.1% and 85.9% for TS and VS, respectively, with no statistical difference found between them. The AcoD of the SM with SP or C increased the production and quality of the biogas, increasing the concentration of CH4 therein.

RESUMO: A co-digestão anaeróbia dos resíduos é uma alternativa para aumentar a produção de metano (CH4) e melhorar a qualidade do biofertilizante. Este estudo teve como objetivo avaliar a melhor condição de coDA dos dejetos de suínos (DS), acrescidos de batata doce (BD) ou mandioca (M) em diferentes inclusões, em biodigestores semi-contínuos. Inicialmente, utilizando biodigestores batelada, foi realizado ensaio de coDA dos DS com BD ou M, adotando-se as relações C/N iguais a 10/1, 13/1, 17/1, e 22/1. Com base nos resultados elegeu-se a relação C/N de 10/1, como sendo a proporção que apresentou maiores reduções de sólidos voláteis (SV) e produções específicas de biogás. A partir destes resultados, efetivou-se o experimento em biodigestores semi-contínuos, composto por três tratamentos (controle (DS), BD 10/1 e M 10/1) com cinco repetições cada, e tempo de retenção hidráulica (TRH) de 30 dias. Foram mensuradas as reduções de sólidos totais (ST) e sólidos voláteis (SV), produções de biogás e CH4, alcalinidade e acidez volátil. O tratamento controle diferiu dos demais e apresentou menor desempenho no rendimento de biogás (752 LN.kgSVadic -1), CH4 (449 LN.kgSVadic -1) e concentração de CH4 (59,7%). Os tratamentos em coDA (BD e M) não diferiram entre si para o rendimento de biogás (901 e 883 LN.kgSVadic -1) e CH4 (590 e 547 LN.kgSVadic -1), no entanto diferiram na concentração de CH4 (65,5 e 61,9%, respectivamente). Os tratamentos apresentaram médias gerais de reduções de 76,1 e 85,9% para ST e SV, respectivamente, não sendo encontrada diferença estatística entre eles. A coDA do DS com BD ou M aumentou a produção e a qualidade do biogás, elevando a concentração de CH4 no mesmo.

In vitro study of the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes / 中华烧伤杂志

Objective@#To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro.@*Methods@#The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent data t test, one-way analysis of variance, least significant difference t test, and Bonferroni correction.@*Results@#(1) Compared with those of normal control group, the protein expressions of LC3Ⅱ and ATP6V1E1 in the whole protein of cells of sugar-free serum-free 1.0, 3.0, and 6.0 h groups were significantly increased (t=12.16, 4.05, 4.82, 11.64, 3.29, 8.37, P<0.05 or P<0.01). Compared with that of normal control group, the protein expression of autophagy-related protein 5 in the whole protein of cells of sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups was significantly increased (t=6.88, 10.56, 5.76, 9.91, P<0.05 or P<0.01). (2) Compared with 1.03±0.08 of normal control group, the lysosomal acidification level in the cells of sugar-free serum-free 3.0 group (2.92±0.30) was significantly increased (t=6.01, P<0.01). (3) There was no statistically significant difference in the overall comparison of protein expression of human antigen R in the whole protein of cells among the 5 groups (F=1.09, P>0.05). Compared with that of normal control group, the protein expression of human antigen R in the cytoplasm protein of cells was significantly increased in sugar-free serum-free 1.0, 3.0, and 6.0 h groups (t=43.05, 11.07, 5.39, P<0.05 or P<0.01), while the protein expression of human antigen R in the nucleus protein of cells was significantly decreased in sugar-free serum-free 3.0 and 6.0 h groups (t=11.18, 12.71, P<0.01). (4) Compared with that of simple control siRNA group, the protein expression of human antigen R in the whole protein of cells of simple HuR-siRNA1 group and simple HuR-siRNA2 group was significantly decreased (t=4.82, 4.44, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the protein expression of human antigen R in the whole protein of cells of sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group was significantly decreased (t=4.39, 6.27, P<0.05). (5) Compared with those of sugar-free serum-free+ control siRNA group, the distribution of human antigen R in the cytoplasm of cells and its expression level were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group (t=10.13, P<0.01). Compared with 1.00±0.06 of sugar-free serum-free+ control siRNA group, the lysosomal acidification level (0.73±0.06) in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=3.28, P<0.01). (6) Compared with those of sugar-free serum-free+ control siRNA group, the protein expressions of cathepsin D in the whole protein of cells, human antigen R in the cytoplasm protein of cells, and ATP6V1E1 in the whole protein of cells were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group (t=4.16, 3.99, 4.81, 5.07, 11.68, 12.97, P<0.05 or P<0.01). (7) Compared with that of normal control group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free 3.0 h group was significantly increased (t=5.51, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=5.97, P<0.05).@*Conclusions@#After sugar-free serum-free treatment in vitro, the autophagy in mouse primary cardiomyocytes is activated, the lysosomal acidification is enhanced, and the expression of human antigen R in cytoplasm is increased. Human antigen R function is activated and involved in maintaining lysosomal acidification during autophagy in mouse cardiomyocytes.

Consideraciones éticas y ambientales en el proceso de acidificación oceánica / Ethical and Environmental Considerations in the Process of Ocean Acidification / Considerações éticas e ambientais no processo de acidificação oceânica

Resumen La acidificación oceánica es un problema creciente que afecta el medio ambiente global, cuyas repercusiones son detectables ahora, que ponen en riesgo el recurso hídrico más extenso del planeta e influyen en los cambios climáticos que se pueden documentar en todo el planeta. El presente artículo analiza la protección del medio marino como una medida para asegurar a las generaciones futuras un ambiente sano, que les garantice una mejor forma de vida. Se exponen los diferentes principios que rigen a la bioética, y se establece su relación con los procesos de desarrollo sostenible y el incremento de la acidificación que ocurre en el océano y que afecta a una gran cantidad de comunidades de escasos recursos a nivel mundial.

Abstract Ocean acidification is a growing problem that affects global environment. Its effects are detectable now, putting the most extensive water resource on the planet at risk and influencing the climate changes that can be documented across the globe. This paper analyzes the protection of the sea as a measure to ensure a healthy environment for future generations, guaranteeing a better way of life for them. We present the different principles governing bioethics, and we establish their relationship with the processes of sustainable development and the increase of acidification in the ocean, which affects a large number of communities of scarce resources in the world.

Resumo A acidificação oceânica, cujas repercussões são detectáveis agora, é um problema crescente que afeta o meio ambiente global, que coloca em risco o recurso hídrico mais extenso do planeta e influencia nas mudanças climáticas que podem ser documentadas em todo o mundo. O presente artigo analisa a proteção do meio marinho como uma medida para assegurar um ambiente saudável às futuras gerações, que lhes garanta uma melhor forma de vida. Os diferentes princípios que regem a bioética são expostos e é estabelecida sua relação com os processos de desenvolvimento sustentável e o aumento da acidificação que ocorre no oceano e que afeta grande quantidade de comunidades com recursos escassos.

Chemical characterization, texture and consumer acceptability of yogurts supplemented with quinoa flour

Abstract Quinoa can be used as a functional ingredient in food formulations. The aim of this study was to evaluate the effects on proximate composition, stability during storage, texture and consumer acceptability of yogurts supplemented with quinoa flour at 1, 3 and 5 g 100 mL-1. A product without supplementation was used as control. Products were assessed for moisture, carbohydrates, proteins, fats, total dietary fibre (TDF), ashes and minerals. The pH, acidity and syneresis of yogurts were measured after 1, 7, 14 and 21 days of storage and a Texture Profile Analysis (TPA) was carried out. Applying hedonic scale, 102 consumers analyzed the overall acceptability, color, texture, flavor and aroma of yogurts. Supplemented products showed significant higher protein, carbohydrate and fat contents. Hardness and adhesiveness showed a negative association whereas a positive one was found between springiness and cohesiveness. Yogurt is not necessarily the adequate matrix for hauling quinoa compounds since the addition of greater amounts of 1 g 100 mL-1 quinoa flour had undesirable effects on gel stability (syneresis and increases in total acidity) and consumer acceptability.

Efecto de la temperatura y la acidificación en larvas de Strombus gigas (Mesogastropoda: Strombidae) / Effect of temperature and acidification in Strombus gigas (Mesogastropoda: Strombidae) larvae

ResumenEl aumento de las emisiones de CO2 produce calentamiento y reducción del pH en los océanos, lo cual puede generar efectos negativos en muchos organismos marinos, particularmente en aquellos con estructuras calcáreas (i.e. moluscos), afectando principalmente a sus estadios larvarios (Le Moullac et al., 2016), en este sentido, se estudió a S. gigas, gasterópodo de importancia comercial en el mar Caribe, con el fin de conocer el efecto de la temperatura y la acidificación en el desarrollo, el crecimiento, la mortalidad y la calcificación durante su fase larvaria. Se realizó un cultivo larvario empleando cuatro tratamientos de temperatura y pH (control = 28 °C - pH 8.1, T1 = 28 °C pH 7.6, T2 = 31 °C pH 8.1 y T3 = 31 °C - pH 7.6) por triplicado. La eclosión se registró al inicio del experimento (No. de huevos - No. de larvas nacidas), por otra parte, el desarrollo de órganos, el crecimiento de la concha y la mortalidad se evaluaron a través del tiempo. La calcificación fue estudiada mediante análisis EDX y Raman para larvas de 30 días de edad. Se observó que el desarrollo y el crecimiento de órganos fue mayor a 31 °C (talla inicial = 230 ± 4.12 a 313.27 ± 11.34 µm, talla final = 829.50 ± 11.33 a 1 054.50 ± 11.13 µm; para T1 y T2, respectivamente), mismo patrón se presentó para el tiempo de eclosión (18 hr) y la tasa de mortalidad (~ 57 %). La proporción de calcio (% wt) fue similar entre tratamientos (de 34.37 ± 10.05 a 37.29 ± 16.81 % wt). El análisis Raman mostró aragonita para las conchas de todas las condiciones experimentales, con valores más altos en el control (1 039.54 ± 780.26 a.u.). Calcita solo se detectó en los tratamientos de 31 °C (174.56 ± 127.19 a.u.), mientras que, a menor pH (7.6), la intensidad de aragonita y calcita fue menor. En conclusión, S. gigas podría adaptarse a escenarios futuros de temperatura y acidificación, sin embargo, puede verse afectado durante los procesos de biomineralización de la concha.

AbstractThe increase in CO2 emissions produces heating and reduced pH in the oceans, which may have negative effects on many marine organisms. This is particularly important for those with calcified structures such as the molluscs and their larval stages. We studied Strombus gigas larvae, a gastropod of commercial importance in the Caribbean Sea, in order to know the effect of water temperature and acidification on their development, growth, mortality and calcification during the larval period. A larval culture with triplicate samples was carried out employing four treatments of temperature and pH (Control = 28 °C - pH 8.1, T1 = 28 °C - pH 7.6, T2 = 31 °C - pH 8.1 and T3 = 31 °C - pH 7.6) in August 2015. We registered hatching (No. of eggs - No. of larvae hatched) and organs development, while shell growth and mortality ratio were evaluated over time. Shell calcification was studied in 30 days old larvae using EDX and RAMAN analysis. Our results showed that organs development and shell growth were higher at 31 °C treatments (initial size of 230 ± 4.12 to 313.27 ± 11.34 µm, and final size from 829.50 ± 11.33 to 1 054.50 ± 11.13 µm; from T1 to T2 respectively), and the same pattern was recorded for hatching time (18 hr) and mortality rate (~ 57 %). The Calcium proportion (% wt) was similar between treatments (34.37 ± 10.05 to 37.29 ± 16.81 % wt). Shell Raman analysis showed aragonite in all experimental treatments, with the highest values in the control (1 039.54 ± 780.26 a.u.). Calcite was detected only in 31 °C treatments (174.56 ± 127.19 a.u.), while less intensity of aragonite and calcite were registered at pH 7.6. In conclusion, S. gigas could be adapted to ocean future predictions, however, shell biomineralization processes can be affected.

Possible mechanism of cellular acidification in epilepsy termination / 国际药学研究杂志

Epilepsy,especially the status epilepticus(SE),is a kind of common disease which is seriously dangerous for peo-ple′s health. At present,many researchers are focusing on how the epilepsy seizures happen and maintain,but know little about how it stops. It was found that appropriate cell acidification played an important role in patients to slow down or inhibit epilepsy-like seizures in the early 20th century. Recently,more and more researches have confirmed that cell acidification can induce the termination of epi-leptic seizure caused by disbalance of inhibitory and excitatory transmitters of nerve,opening of acid-sensing ion channels and so on. Therefore,these will likely become the new direction to explore new drugs or methods for epileptic treatment in the future.

Deep sequencing analysis on transcriptomes of Edwardsiella tarda regulated by Eha following acidification / 中国人兽共患病学报

Our studies tried to demonstrate Eha (Et haemolysin activator) could regulate the resistance of the bacterium against acidification to survive in the macrophage and explain its underlying molecular mechanism.When the bacteria infected the macrophages at time intervals,intracellular survival rate in bafilomycin-treated macrophages was higher than that with untreated cells,and the rate of wild type ET 13 was higher than that of its eha mutant,respectively (P＜0.05).The survival rate of the wild type was higher than that of the mutant under acid treatment (P＜0.05).To determine the conditions that induced the highest eha expression,we constructed a pMP220-Peha LacZ plasmid and determined the lacZ expression under different conditions.After exposure of pH6.3 medium for 2 h time,we performed the whole transcriptomic profiles of the wild type and mutant by RNA-sequencing.We identified 147 differentially-expressed genes ([log2 ratio| ≥1),113 and 34 of which were significantly up-and down-regulated,respectively in the mutant,comparing with the wild type.These findings were validated by qRT-PCR.GO functional analysis revealed that these genes were divided into 25 categories,including the bacterial catalysis,cellular composition,combination,localization,metabolism,processing,and transportation.Based on the KEGG database,these genes were distributed in 55 pathways,such as two-component system,ABC transporters,and microbial metabolism in diverse environments.Overall,Eha is an important regulator to affect all kinds of target genes and pathways for E.tarda to adapt to an acid environment.These results could be helpful for further investigations of the mechanisms by which E.tarda survives in macrophages.

Optimization of self-nanoemulsifying formulations for weakly basic lipophilic drugs: role of acidification and experimental design

ABSTRACT Formulators face great challenges in adopting systematic approaches for designing self-nanoemulsifying formulations (SNEFs) for different drug categories. In this study, we aimed to build-up an advanced SNEF development framework for weakly basic lipophilic drugs, such as cinnarizine (CN). First, the influence of formulation acidification on CN solubility was investigated. Second, formulation self-emulsification in media with different pH was assessed. Experimentally designed phase diagrams were also utilized for advanced optimization of CN-SNEF. Finally, the optimized formulation was examined using cross polarizing light microscopy for the presence of liquid crystals. CN solubility was significantly enhanced upon external and internal acidification. Among the various fatty acids, oleic acid-based formulations showed superior self-emulsification in all the tested media. Surprisingly, formulation turbidity and droplet size significantly decreased upon equilibration with CN. The design was validated using oleic acid/Imwitor308/Cremophor El (25/25/50), which showed excellent self-nanoemulsification, 43-nm droplet size (for CN-equilibrated formulations), and 88 mg/g CN solubility. In contrast to CN-free formulations, CN-loaded SNEF presented lamellar liquid crystals upon 50% aqueous dilution. These findings confirmed that CN-SNEF efficiency was greatly enhanced upon drug incorporation. The adopted strategy offers fast and accurate development of SNEFs and could be extrapolated for other weakly basic lipophilic drugs.

Efeito da composição de bactérias láticas e da suplementação do soro de leite na cinética de acidificação, crescimento celular e produção de nisina / Effect of lactic acid bacteria composition and the supplementation of milk whey on acidification kinetics, cell growth and nisin production

As bactérias ácido-láticas (BAL) são micro-organismos que auxiliam nas características organolépticas, funcionais e de bioconservação de produtos fermentados. A utilização do soro de leite como meio de cultivo natural enaltece o conceito da produção de biomoléculas de alto valor agregado, como bacteriocinas, já que é um subproduto gerado por indústrias de laticínios e considerado um agente poluidor. A inulina é um ingrediente prebiótico que promove seletivamente o crescimento de culturas probióticas. Nesse âmbito, o objetivo deste estudo foi avaliar o efeito da composição da cultura de Lactococcus lactis (LL) em cocultura com Streptococcus thermophilus (ST) e da suplementação da base de soro de leite com inulina: (i) nos parâmetros cinéticos de acidificacão, (ii) no crescimento celular, (iii) na viscosidade do produto e (iv) na atividade antimicrobiana da nisina. A fermentação do soro de leite com Lactococcus lactis em cocultura com Streptococcus thermophilus proporcionou a maior taxa de acidificação (Vmax=7,93x10-3 upH/min), assim como apresentou o menor tempo para atingir a velocidade máxima de acidificação (Tvmax=1,13 h). A adição de 2% de inulina ao soro de leite fermentado pela cocultura binária fez com que o tempo para completar o cultivo fosse o mais curto (TpH4,5=4,43 h) quando comparado aos demais ensaios. Quanto ao crescimento celular, pode-se observar que a inulina não afetou significativamente a contagem microbiológica, quando as cepas ST e LL foram utilizadas separadamente no processo fermentativo. Em particular, a adição de 4% de inulina reduziu em 1,2 LogUFC/mL e 0,92 LogUFC/mL a contagem de ST e LL (em monocultura), respectivamente. Por outro lado, em coculturas binárias (ST-LL), percebeu-se ganho na contagem microbiológica nos ensaios que receberam suplementação do ingrediente prebiótico, ou seja, quando adicionados 2% e 4% de inulina, houve aumento de 1 LogUFC/mL e de 1,34 LogUFC/mL na contagem de ST, respectivamente. No caso da cepa LL em cocultura com ST, a suplementação de 2% e 4% do prebiótico aumentou em 0,31 LogUFC/mL e 0,75 LogUFC/mL, respectivamente. A concentração de ácido lático também foi mais elevada nos cultivos realizados com a cocultura binária, sendo 4,56 g/L (na ausência de inulina), 5,28 g/L (com adição de 2% de inulina) e 5,71 g/L (com suplementação de 4% de inulina). A viscosidade foi influenciada tanto pela adição de inulina como pelo efeito sinérgico da cocultura, sendo que o maior valor (7,38 mPas) foi obtido pela cocultura ST-LL e pela adição de 4% do ingrediente prebiótico. Quanto à produção de nisina, observou-se que, no cultivo em cocultura (ST-LL), a concentração de 2% de inulina aumentou em 102% a atividade antimicrobiana quando comparada com a cultura pura LL. Vale ressaltar que ambas as cepas satisfizeram os requisitos tecnológicos relativos à produção de laticínios funcionais

Lactic acid bacteria (LAB) are microorganisms that help in the organoleptic and functional characteristics and in the biopreservation of fermented products. The use of milk whey as a culture medium extols the concept of the production of high value-added biomolecules, such as bacteriocins, since it is a by-product generated by the dairy industry and considered a pollutant. Inulin is a prebiotic ingredient that promotes selectively the growth of probiotic cultures. In this context, the aim of this study was to evaluate the effect of culture composition Lactococcus lactis (LL) in co-culture with Streptococcus thermophilus (ST) and the supplementation of milk whey with inulin on: (i) the acidification kinetic parameters, (ii) the cell growth, (iii) the product viscosity, and (iv) the antimicrobial activity of nisin. The fermentation of milk whey by Lactococcus lactis in coculture with Streptococcus thermophilus provided the highest acidification rate (Vmax = 7.93x10-3 upH/min) and the shortest time to reach the maximum acidification rate ( TVmax = 1.13 h). The addition of 2% inulin in the binary coculture binary led to the shorter time to complete the fermentation (TpH4,5 = 4.43) compared to the other tests. With regard to cell growth, it can be observed that the addition of inulin did not affect the microbiological count of pure cultures of ST and LL strains in the fermentation process. In particular, the addition of 4% inulin reduced by 1.2 Log CFU/mL and 0.92 Log CFU/mL the counts of ST and LL (monoculture), respectively. In the other hand, the binary co-cultures cultivations (ST-LL) with the addition of 2% and 4% inulin increased by 1 LogCFU/mL and 1.34 Log CFU/mL in the case of the ST counts and 0.31 log CFU/mL and 0.75 log CFU/mL the counts of LL, respectively. Lactic acid concentration was higher in cultivations carried out by binary cocultures, thus being 4.56 g/L (in the absence of inulin), 5.28 g/L (with addition of 2% inulin) and 5.71g/L (supplemented with 4% inulin). The viscosity was influenced by the addition of prebiotic ingredient and by the synergistic effect of binary coculture, being the highest value (7.38 mPas) obtained by the addition of 4% inulin. Finally, as regards the production of nisin noted that in the binary coculture cultivations (ST-LL), the concentration of 2% inulin increased at 102% the antimicrobial activity when compared to the pure culture LL. It is worth mentioning that both strains met the technological requirements as regards the production of functional dairy products

Enraizamento de estacas de genótipos de Camellia sinensis L. em meio ácido, presença de alumínio e ácido indolbutírico / Rooting of Camellia sinensis L. in acid medium, the presence of aluminum and indolebutyric acid

RESUMO Experimentos foram conduzidos para avaliar o efeito do meio ácido e do alumínio, assim como para determinar a concentração mais apropriada de ácido indolbutírico (AIB) para o enraizamento de estacas de diferentes genótipos de Camellia sinensis L (planta de chá). Para tal, foram coletados ramos de plantas-matrizes em Pariquera-Açu-SP, Brasil, no inverno de 2012 e preparadas estacas semi-lenhosas, contendo uma gema e uma folha, que foram mantidas em viveiro com 70% de sombreamento. A irrigação do substrato foi feita com água e soluções contendo ácido fosfórico e sulfato de alumínio a pH 5,5, 4,5; 3,5. No primeiro experimento, o delineamento experimental foi o inteiramente casualizado em esquema fatorial 3 x 7, três genótipos (F 15, IAC 259 e Comum) e sete condições diferentes de enraizamento (vermiculita a pH 6,5, vermiculita acidificada com ácido fosfórico a pH 3,5; 4,5 e 5,5 ou vermiculita acidificada com sulfato de alumínio a pH 3,5; 4,5 e 5,5). No segundo experimento, o tratamento que promoveu o maior enraizamento no primeiro experimento (vermiculita acidificada com sulfato de alumínio a pH 3,5) foi combinado ao tratamento com AIB. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial 3 x 6, sendo três genótipos e seis concentrações de AIB (0, 2000, 4000, 6000, 8000 e 1000 mg L-1). A vermiculita acidificada com sulfato de alumínio a pH 3,5, combinada a aplicação de 10000 mg L-1 de AIB por 30 segundos, foi o tratamento mais adequado para o enraizamento de estacas semi-lenhosas dos genótipos F15, IAC 259 e Comum.

ABSTRACT The experiments were carried out to evaluate the effect of medium acidic and aluminum, as well as determine the most suitable concentration of indolebutyric acid (IBA) for rooting cuttings of different genotypes of Camellia sinensis L (tea plant). Such, stems were collected from mother plants in Pariquera-Açu, São Paulo state, Brazil, in winter 2012 and prepared semi-hardwood cuttings, with one bud and one leaf, which were kept in a nursery with 70% of shading. Irrigation substrate was taken with water and solutions containing phosphoric acid and aluminum sulfate at pH 5.5, 4.5; 3,5. In the first experiment, the experimental design was completely randomized in a factorial scheme 3x7, three genotypes (F 15, IAC 259 and Comum) and seven different conditions of rooting (vermiculite at pH 6.5, acidified with phosphoric acid or aluminum sulfate vermiculite at pH 3.5, 4.5 and 5.5). In the second experiment, the treatment that promoted the highest rooting in the first experiment (vermiculite acidified with aluminum sulfate at pH 3.5) was combined with treatment with IBA The experimental design was completely randomized in a factorial scheme 3x6, three genotypes and six concentrations of IBA (0, 2.000, 4.000, 6.000, 8.000 and 10.000 mg L-1). Vermiculite acidified with aluminum sulphate to pH 3.5, combined application of 10.000 mg L-1 IBA for 30 seconds was the most appropriate treatment for cutting propagation of genotypes F15, IAC 259 and Comum. Vermiculite acidified with aluminum sulfate at pH 3.5, combined treatment with 10,000 mg L-1 IBA for 30 seconds, was the most suitable conditions for the rooting of cuttings Camellia sinensis L.

Effects of BAPTA-AM on acid-induced autophagy of rat articular chondrocytes and its possible mechanisms / 中国药理学通报

Aim To observe the effect of BAPTA-AM on extracellular acid-induced autophagy in rat articular chondrocytes and its possible mechanisms.Methods Rat articular chondrocytes were isolated from Sprague-Dawley rats and incubated with different pH medium. The states of autophagy were examined by acridine or-ange (AO ) staining .Moreover,the expressions of LC3 ,Beclin-1 ,ULK1 ,CaMKKβ,AMPK and mTOR were detected using Western blot or quantitative real-time PCR (qRT-PCR ). Intracellular calcium ([Ca2+]i )was analyzed by a Ca2+-imaging method. Results Compared with pH 6.0 group,BAPTA-AM could significantly decrease the activation of autophagyinduced by acid exposure,and the expressions of autophagy markers including LC3 Ⅱ,Beclin1 and ULK1were also decreased,accompanied with reduced acidinduced [Ca2 +]i influx,decreased proteins expressionof CaMKKβand phosphorylatedAMPK,and increasedphosphorylation of mTOR.Conclusion BAPTAAMcan significantly restrain acidinduced autophagy in ratarticular chondrocytes,the mechanism of which may beassociated with decreased Ca2 + influx.

Modulation of phagolysosome maturation by bacterial tlyA gene product.

The pathogenic traits of TlyA proteins of Mycobacterium tuberculosis are not known. Expressions of TlyA in bacteria that do not express endogenous TlyA adhere better to RAW264.7 macrophages and get phagocytosed efficiently. The internalized bacteria avoid acidification to the extent of >65% in the case of both TlyA-expressing E. coli and M. smegmatis. Consistent with this observation, we have observed decreased co-localizaton of Lysosomal Membrane Associated Protein-1 (~35%), Early Endosomal Antigen-1 (~34%), Rab5 (~30%) and Rab7 (~35%) and enhanced colocalizaton of Rab14 (~80%) on both TlyA-expressing bacteria as well as on TlyA-coated latex beads. These results suggest that the mycobacterial TlyA, in general, can modulate phagolysosome maturation pathway immediately after entry into macrophages, while other important molecules may aid the bacterium for long-term, intracellular survival at later point of time.

Influence of water pH in oral changes caused by cadmium poisoning: an experimental study in rats / Influência do pH da água nas alterações bucais causadas pela intoxicação por cádmio: um estudo experimental em ratos

Objective: This study aims to evaluate the effect of the pH of drinking water in the oral changes caused by cadmium poisoning. Material and method: Ninety male Wister rats were divided into the following six groups: A - 15 rats were given cadmium chloride solution (400 mg/L) in drinking water with a neutral pH (pH 7.0); B - 15 rats received cadmium chloride solution (400 mg/L) in drinking water with an acidic pH (pH 5.0); C - 15 rats were treated with a cadmium chloride solution (400 mg/L) in drinking water with a basic pH (pH 8.0); D - 15 rats received drinking water with an acidic pH (pH 5.0); E - 15 rats were given drinking water with a basic pH (pH 8.0); F - 15 rats received water with a neutral pH (pH 7.0). All animals were sacrificed six months after the beginning of the experiment. A biopsy of the buccal mucosa, tongue and salivary gland of each animal was taken for microscopic analysis. Result: No changes were observed in the buccal mucosa, tongue mucosa or salivary glands in any of the groups. Conclusion: Drinking water that contains a high concentration of cadmium with differing pH levels demonstrated no damage to the oral mucosa and salivary glands of male Wistar rats. .

Objetivo: Avaliar o efeito do pH da água de beber nas alterações bucais provocadas pela intoxicação por cádmio. Material e método: Foram utilizados 90 ratos Wistar, adultos, machos, divididos em 6 grupos: A - 15 ratos que receberam solução de cloreto de cádmio (400 mg/L) na água de beber com pH neutro (pH 7,0); B -15 ratos que receberam solução de cloreto de cádmio (400 mg/L) na água com pH ácido (pH 5,0); C - 15 ratos, os quais receberam solução de cloreto de cádmio (400mg/L) na água com pH básico (pH 8,0). D - 15 ratos que receberam água com pH ácido (pH 5,0); E - 15 ratos que receberam água com pH básico (pH 8,0); F -15 ratos que receberam água com pH neutro (pH 7,0). Todos os animais foram eutanasiados 6 meses após o início do experimento. Foram retirados fragmentos da mucosa jugal, língua e glândula salivar de cada animal para análise microscópica. Resultado: Não foram observadas alterações na mucosa jugal, mucosa da língua ou nas glândulas salivares em nenhum dos grupos avaliados. Conclusão: Mesmo em alta concentração o cádmio adicionado à água de beber não mostrou causar dano a mucosa bucal ou às glândulas salivares, independente do pH da água. .

Regulation and significance of acidification of plasma membrane and intracellular membranes / 医学研究生学报

Acidic environment of organelles of eukaryotic cells plays an important role in endocytosis, the secretion of lysoso-mal enzymes and other physiological activities.V-ATPase (vacuolar ATPases) and CLC (chloride channel) family are widely distribu-ted and present in virtually all eukaryotic cells in intracellular membranes and in the plasma membrane.They are responsible for the a-cidification of the vesicular interior.In this paper, the distribution, structure, mechanism of action and physiological and pathological significance of V-ATPase and CLC protein are discussed.

Synaptic vesicle cycling is not impaired in a glutamatergic and a cholinergic synapse that exhibit deficits in acidification and filling

The purpose of the present work was to investigate synaptic vesicle trafficking when vesicles exhibit alterations in filling and acidification in two different synapses: a cholinergic frog neuromuscular junction and a glutamatergic ribbon-type nerve terminal in the retina. These synapses display remarkable structural and functional differences, and the mechanisms regulating synaptic vesicle cycling might also differ between them. The lipophilic styryl dye FM1-43 was used to monitor vesicle trafficking. Both preparations were exposed to pharmacological agents that collapse ΔpH (NH4Cl and methylamine) or the whole ΔµH+ (bafilomycin), a necessary situation to provide the driving force for neurotransmitter accumulation into synaptic vesicles. The results showed that FM1-43 loading and unloading in neuromuscular junctions did not differ statistically between control and experimental conditions (P > 0.05). Also, FM1-43 labeling in bipolar cell terminals proved highly similar under all conditions tested. Despite remarkable differences in both experimental models, the present findings show that acidification and filling are not required for normal vesicle trafficking in either synapse.

O objetivo do presente trabalho foi investigar o tráfego de vesículas sinápticas quando estas apresentam alterações no armazenamento de neurotransmissores e acidificação em duas distintas sinapses: a junção neuromuscular colinérgica de rãs versus o terminal nervoso glutamatérgico do tipo ribbon em céulas bipolares da retina. Essas sinapses exibem notáveis diferenças estruturais e funcionais e os mecanismos de regulação de ciclo das vesículas sinápticas podem ser diferentes entre eles. Para monitorar o tráfego de vesícula, foi utilizado o marcador lipofílico FM1-43. Ambas as preparações foram expostas a agentes farmacológicos que provocam o colapso de ΔpH (NH4Cl e metilamina) ou de todo ΔµH+ (bafilomicina), gradientes necessários para o acúmulo de neurotransmissores em vesículas sinápticas. Nossos resultados demonstram que a marcação e desmarcação de FM1-43 nas junções neuromusculares não foi estatisticamente diferente entre as diversas condições experimentais (P > 0,05). Além disso, a marcação de FM1-43 em terminais sinápticos de células bipolares foram bastante semelhantes em todas as condições testadas. Apesar das diferenças marcantes em ambos os modelos experimentais, nossos achados demonstram que a acidificação e o preenchimento de vesículas sinápticas não são necessários para o tráfico normal da vesícula nas sinapses estudadas.

Digestibilidade de nutrientes e balanço de Ca e P em suínos recebendo dietas com ácido butírico, fitase e diferentes níveis de cálcio / Nutrient digestibility and Ca and P balance in pigs receiving butyric acid, phytase and different calcium levels

Estudou-se o uso do ácido butírico (AB) e da fitase em dietas de suínos na fase de crescimento, variando no nível de cálcio. O experimento foi subdivido no tempo em dois períodos de 17 dias, sendo três de adaptação e 14 de mensurações. Em cada período, foram utilizados 16 suínos machos castrados, com peso de 24,6±0,7kg no primeiro e 43,2±1,77kg no segundo período. As dietas diferiam no nível de cálcio (0,5 ou 0,72 por cento), de AB (0 ou 0,3 por cento de butirato de sódio 84 por cento) e de fitase (0 ou 500 FTU kg-1, fitase de origem bacteriana derivada de Escherichia coli). O delineamento foi em blocos casualizados (períodos), em decomposição fatorial 2x2x2, com quatro repetições. Foram avaliadas a digestibilidade aparente dos nutrientes e da energia bruta e o balanço de Ca e P. O AB melhorou a digestibilidade da proteína bruta, mas, de forma individual ou em combinação com a fitase, não aumentou a retenção de minerais. A fitase aumentou a retenção de P, reduzindo sua excreção fecal e urinária. O menor nível de Ca na dieta proporcionou maior retenção de Ca e menor retenção de P, em decorrência do aumento da excreção de P na urina (P<0,0001). Apesar de ter melhorado a digestibilidade protéica, o AB não aumentou a retenção de minerais, nem teve efeito aditivo ao uso de fitase, enquanto que a fitase apresentou efeito positivo para retenção de P.

It was studied the use of butyric acid (BA) and phytase in growing pigs diets, varying calcium level. The experiment was divided into two periods of time, of 17 days, with 3 days of adaptation and 14 of measurements. In each period, it was used 16 barrows weighing 24.6±0.7kg in the first and 43.2±1.77kg in the second. Diets were different in calcium level (0.5 or 0.72 percent), AB (0 or 0.3 percent sodium butyrate 84 percent) and phytase (0 or 500FTU kg-1 phytase of bacterial origin derived from Escherichia coli). The experimental design was in randomized blocks (periods), decomposed in 2x2x2 factorial, with four replications. It was evaluated the apparent digestibility of nutrients, gross energy and balance of Ca and P. AB improved crude protein digestibility, but individually or in combination with phytase did not increase minerals retention. Phytase increased P retention by reducing urinary and fecal excretion. The lowest diet calcium level provided higher Ca retention and lower P retention as a result of increased P excretion in urine (P<0.0001). Despite improving protein digestibility, AB did not increase mineral retention and didn't have an additive effect with the use of phytase, while phytase had positive effect on retention of P.

Atributos químicos relacionados ao processo de sulfurização em solos construídos após mineração de carvão / Chemical attributes of constructed soils after surface coal mining related to sulphurization process

A mineração de carvão a céu aberto envolve a remoção superficial do solo e de camadas geológicas, os quais devem ser repostos visando à reabilitação das áreas mineradas. Essa obra expõe o carvão às condições oxidativas do ar e altera as características originais do solo e da paisagem. Como consequência, o processo de sulfurização pode ser ativado nos solos construídos em razão da oxidação da pirita, condicionando alterações químicas e mineralógicas. Nesse contexto, o estudo objetivou avaliar a evolução temporal de solos construídos há 24 (SA-24) e dois (SA-2) anos, em duas áreas mineradas e reconstruídas no Município de Minas do Leão, Rio Grande do Sul (RS), tendo como base atributos químicos relacionados com o processo de sulfurização. Para tanto, foram determinados o pH, a condutividade elétrica (CE), os teores de bases (SB) e de alumínio (Al3+) trocáveis, a acidez potencial (H+Al), os teores totais de Al, Fe e Si e o teor de sulfato solúvel (S-SO4(2-)) e calculadas a capacidade de troca de cátions (CTC), a saturação por bases (V) e a saturação por alumínio (m). Os resultados indicaram a atuação do processo de sulfurização nos solos construídos de ambas as áreas reabilitadas. A CE e os teores de S-SO4(2-) foram maiores na SA-2, indicando um estádio de sulfurização mais ativo nos solos dessa área. Na SA-24, a menor CE e os menores teores de S-SO4(2-), de SB e V, bem como a maior saturação por Al e teores de H+Al, sugerem a proximidade do estádio de pós-sulfurização. Elementos como Al e P concentraram-se relativamente nos solos construídos mais antigos. Os solos em ambas as áreas apresentam limitações químicas para o desenvolvimento de vegetação, influindo negativamente para a recuperação das áreas.

Coal stripmining involves the removal of upper soil and geological layers, which must be relocated in a similar way that in the original profile when reconstructing the landscape. As this process changes soil and landscape characteristics, pirite may oxide and change soil chemical and mineralogical characteristics. In this context, the study aimed to evaluate the temporal evolution of soils reconstructed 2 (SA-2) and 24 (SA-24) year ago, in Boa Vista Coal Mining, in Minas do Leão, Rio Grande do Sul, Brazil. Soil characteristics measured were pH, electrical conductivity (EC), bases and aluminium content, potential acidity (H+Al), and contents of Al, Fe, Si, and soluble sulphate. Calculations accounted for cation exchange capacity, base saturation and aluminum saturation. Results indicated occurrence of oxidation processes in both reconstructed areas. Electrical condutivity and soluble phosphate contents were higher in SA-2, indicating a more active sulphurization stage in these soils. In soils of area SA-24, lower EC and smaller contents of soluble sulphate, lower base saturation, as well as higher aluminum saturation and H+Al suggest a more advanced sulphurization process compared to SA-2. Aluminum and P concentrated in older soil profiles. Soils in both areas have chemical limitations for plant growth with adversely affects to recuperation of the area.