Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-¹. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-¹. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.

Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766)

Abstract This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Resumo Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications

Abstract The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by-products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 Umg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

Resumo As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 Umg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These byproducts can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.

Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

Being born and growing up in the Ribeirão Preto and São Luís cohorts

This thematic issue consists of 14 articles derived from studies of the BRISA birth cohort (Ribeirão Preto, State of São Paulo and São Luís, State of Maranhão, northeastern Brazil, a socially and economically less developed region). In these more than 40 years of existence, these cohorts have been able to document the increase in women's education, the improvement of health conditions, the creation of a public Unified Health System (SUS) that provides universal and free access to health care, eradication of hunger, and transition of the nutritional status characterized by a decrease in malnutrition rates and an increase in obesity in Brazil. Particularly in reproductive health, the country experienced a significant drop in fertility, a decrease in maternal and child mortality, and an increase in breastfeeding rates. Universal access to prenatal care and hospital delivery was accompanied by an excessive number of cesareans without clinical indication and early-term births and premature births, largely due to scheduled cesareans. Articles with a longitudinal and transversal methodological approach are presented, using structural equation analysis and propensity score, together with multivariate regressions, which gave a robust analytical treatment to articles in this thematic issue.

Efeito da suplementação de colina protegida no perfil metabólico e intervalo entre parto e concepção de vacas leiteiras / Prepartum supplementation of protected choline on metabolic profile and interval between calving and conception of dairy cows

Durante o periparto, as vacas leiteiras são submetidas a uma grande demanda de energia, ao mesmo tempo em que reduzem sua ingestão de matéria seca. O balanço energético negativo, resultante dessa equação, acarreta severos transtornos metabólicos, à produção e, principalmente, à reprodução. O objetivo do presente estudo foi avaliar o efeito da colina protegida sobre os parâmetros metabólicos, o intervalo entre parto e concepção e a produção de leite em vacas no período de transição. Cinquenta e quatro vacas leiteiras foram divididas em três grupos: controle, suplementação com colina por 10 dias pré-parto (T10) e suplementação com colina por 20 dias pré-parto (T20). Após o parto, foram mensurados os teores de frutosamina, colesterol, ácidos graxos não esterificados (AGNE), beta-hidroxibutirato (BHB), aspartato aminotransferase (AST), gamaglutamiltransferase (GGT) e total de oxidantes (TOS), nos dias 10, 20 e 30. Ainda foram avaliadas produção de leite e intervalo entre parto e concepção. Não houve efeito da suplementação com colina sobre os parâmetros sanguíneos e a produção. O intervalo entre parto e concepção foi menor no grupo T20. A colina suplementada por 20 dias durante o pré-parto melhorou a performance reprodutiva de vacas leiteiras(AU)

During the periparturient dairy cows undergo a large energy demand, at the same time reducing their intake of dry matter. The negative energy balance resulting from this equation leads to severe metabolic disorders in production, and mainly in reproduction. The aim of this study was to evaluate the effect of protected choline on metabolic parameters, reproductive performance, and milk production in cows during the transition period. Fifty-four dairy cows were divided into three groups: control, supplementation with choline for 10 days prepartum (T10) and supplementation with choline for 20 days prepartum (T20). After delivery we measured fructosamine levels, cholesterol, non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), aspartate aminotransferase (AST), gamma glutamyltransferase (GGT), and total oxidant (TOS) on days 10, 20 and 30. We also evaluated milk production and interval between calving and conception. There was no effect of supplementation with choline on blood and production parameters. The interval between calving and conception was lower in the T20 group. Choline supplemented by 20 during the antepartum improved reproductive performance of dairy cows, although it did not change the metabolic profile.(AU)

Efeito do uso de minerais traços (cobre, zinco, selênio e manganês) e vitaminas (A e E) injetáveis sobre o estresse oxidativo e função de neutrófilos de fêmeas da raça Holandesa no período de transição / Effect of use of injected trace minerals (copper, zinc, selenium, and manganese) and vitamins (A and E) on oxidative stress and neutrophil function in females of Hostein cows during transitionl

O objetivo deste estudo foi avaliar o efeito metafilático de minerais traços e vitaminas A e E injetáveis, em parâmetros do estresse oxidativo e na função de neutrófilos em fêmeas da raça Holandesa no período de transição. Sessenta animais foram divididos em dois grupos: animais que receberam minerais traços e vitaminas A e E injetáveis (GMTV) (n= 30) e grupo controle (GC) (n= 30). Não houve diferença significativa entre os grupos nos parâmetros avaliados, porém observou-se diferença significativa entre tratamento e dia para os valores da SOD, com maior atividade dessa enzima em fêmeas GMTV, nas semanas próximas ao parto. Observou-se diferença de dia e para interação tratamento e dia para o TBARS, em que fêmeas GMTV mostraram menores valores de TBARS em todos os momentos, exceto sete dias após o parto. Houve efeito significativo de dia para leucócitos, interação tratamento e dia para neutrófilos e interação tratamento e dia para fagocitose de neutrófilos, em que as fêmeas GMTV apresentaram valores menores de leucócitos e neutrófilos próximo ao parto, além de maior fagocitose de neutrófilos. Pode-se observar que houve melhora no sistema oxidativo e imune de fêmeas GMTV, resposta que provavelmente está relacionada com a administração dos minerais traços e vitaminas A e E.(AU)

This study evaluates the metaphylactic of the subcutaneous administration of a trace minerals and vitamins A and E, on the oxidative stress and neutrophil function in Holsteins cows under the transition period. Sixty females were divided in two groups: group with trace minerals and vitamins (GMTV) (n= 30) and group control (GC) (n= 30). There was no significant difference between those groups; however, we find significant difference between treatment and day for Superoxide dismutase (SOD) values with higher activity of this enzyme in females GMTV on the weeks next to the parturition. Still, there was difference on day and, for interactions between treatment and day for TBARS, were females GMTV showed lower values of TBARS in all moments, except on day seven after the parturition. For leucocytes, there is a significant effect by day, interaction on treatment and day on neutrophils, and interaction treatment and day for neutrophil phagocytosis, were females GMTV showed lower values of leucocytes and neutrophils next to the parturition, and an increase of neutrophils phagocytosis. In summary, cows from the GMTV group had an improvement on the immune and oxidative systems, probably correlated with the administration of this supplement.(AU)

Imágenes en divertículos del tubo digestivo: localizaciones infrecuentes. serie de casos / Imaging in infrequent digestive diverticula: case series

Los divertículos se pueden localizar en todo el tubo digestivo (TD): esófago, estómago, duodeno, yeyuno, íleon, apéndice, colon. Son infrecuentes, salvo en colon. Sus manifestaciones clínicas son inespecíficas, con difícil diagnóstico y mayor riesgo de complicaciones. Se presenta una serie de divertículos digestivos atípicos, mostrando las características imagenológicas multimodalidad y describir los hallazgos claves. Es una revisión retrospectiva en el archivo computacional de nuestra institución. Selección de casos de divertículos de presentación inhabitual por sus características, localización u origen. En esófago los divertículos de Zenker y Killian-Jamieson. En estómago los divertículos gástricos infrecuentes. Los divertículos en intestino delgado tienen baja prevalencia, el más frecuente en duodeno. Los divertículos colónicos pueden tener una localización o presentación atípica. Los divertículos apendiculares y Meckel presentan baja prevalencia. Los divertículos digestivos son infrecuentes, excepto los colónicos. El radiólogo debe estar familiarizado con las diferentes ubicaciones de ellos, para reconocerlos y poder diagnosticarlos.

Diverticula may occur in any segment of the digestive tract: esophagus, stomach, duodenum, jejunum, ileum, appendix, and colon. Its clinical manifestations are nonspecifi which may turn diffiult an early diagnosis, leading to a higher risk of complications. We present a cases serie of atypical digestive diverticula and to describe the fidings on the different imaging techniques. We performed a retrospective review on the imaging computer archives of our institution. The atypical diverticula were selected. Zenker and Killian-Jamieson's diverticulums in esophagus. In stomach ocasionally gastric diverticula. Diverticula in small intestine have a low prevalence, the most common location is duodenum. Colonic diverticula may present a atypical location or presentation. The appendicular and Meckel diverticuli are uncommon. The digestive diverticula are uncommon, except the colonic ones. The radiologist must be familiar with it's different locations in order to be able to recognize it and diagnose it properly.

Distribution of selenium in sheep treated with dipheny diselenide / Distribuição do selênio em ovinos tratados com disseleneto de difenila

The aim of the present study was to report the in vivo distribution of selenium in sheep. For this, animals were allocated into two groups (control group and treated group) and kept in metabolic cages for a period of 37 days. The treated group received a single dose (6µmol/kg) of Diphenyl Diselenide, intravenously. Plasma and erythrocytes samples were collected at different times. Adipose tissue, muscles (latissimusdorsi, semitendinosus, and supra-scapular) heart, liver, lung, kidney, intestine and brain were sampled at 30 days post-treatment, in order to determine the selenium concentration. The results demonstrated that the selenium, from the Diphenyl Diselenide group, was higher in erythrocytes (4.8mg/L, six hours post-treatment) when compared with the control sheep. The deposition of selenium occurred in the liver (7.01µg/g), brain (3.53µg/g) and kidney (2.02µg/g). After 30 days of a single intravenous injection of Diphenyl Diselenide, liver was the main organ of selenium deposition.(AU)

O objetivo do presente estudo foi investigar a distribuição in vivo do selênio em ovinos. Para isso, os animais foram distribuídos em dois grupos (grupo controle e grupo tratado) e mantidos em gaiolas metabólicas por um período de 37 dias. O grupo tratado recebeu uma dose única (6µmol/kg) de disseleneto de difenila, por via intravenosa. As amostras de plasma e de eritrócitos foram recolhidas em momentos diferentes. Tecido adiposo, músculos (latissimus dorsi, semitendinoso e supraescapular) coração, fígado, pulmão, rim, intestino e cérebro foram amostrados aos 30 dias pós-tratamento, a fim de se determinar a concentração de selênio. Os resultados demonstraram que o selênio, do grupo disseleneto de difenila, foi maior em eritrócitos (4,8mg/L, seis horas após o tratamento) quando comparado com o grupo controle. A deposição de selênio ocorreu no fígado (7,01µg/g), cérebro (3,53µg/g) e rim (2,02µg/g). Após 30 dias de uma única injeção intravenosa de disseleneto de difenila, o fígado foi o principal órgão de deposição de selênio.(AU)

Efeito metafilático do zinco injetável sobre parâmetros metabólicos e oxidativos de ovelhas no pós-parto imediato / Metaphylactic effect of injectable zinc on metabolic and oxidative parameters of sheep in the immediate postpartum

The aim of this study was to evaluate a new source of injectable organic zinc (zinc edetate) on the energy and oxidative profile in sheep during the immediate postpartum period. Twenty-six Texel breed animals were previously identified and divided into two experimental groups: the treated group (TG; n= 13) that comprised the animals that received a subcutaneous (SC) injection of 100 mg of zinc edetate (2 mL) fifteen days before the parturition expected date and the control group (CG; n=13) that comprised the animals that received 2mL of physiological solution at the same date of TG. Blood samples were collected on the parturition day for the assessment of serum fructosamine, cholesterol and triglycerides, insulin-like growth factor type 1 (IGF-1), the oxidative stress index (OSi) and blood zinc concentration. In addition to these parameters, the measurement of zinc was made in food given to the animals. There was no difference in metabolic parameters and OSi between the experimental groups (P>0.05), as well as in blood zinc concentrations (P>0.05). The parenteral zinc edentate does not change the energy and oxidative profile of sheep in immediate postpartum.(AU)

Efeito da suplementação com colina protegida sobre parâmetros bioquímicos, produção e reprodução de vacas leiteiras no periparto / Effect of protected choline supplementation on biochemical parameters, production, and reproduction of dairy cows in peripartum

Avaliou-se o efeito da suplementação com colina protegida sobre o perfil energético, as enzimas hepáticas e a reprodução de vacas leiteiras no periparto. Quinze vacas leiteiras foram divididas em dois grupos experimentais: oito receberam 80 gramas de colina protegida por 21 dias no pré-parto e por 40 dias no pós-parto e sete foram consideradas controle. Amostras de sangue foram coletadas nos dias 10, 20, 30 e 60 pós-parto para avaliação dos perfis energético e hepático. Aos 60 dias pós-parto, realizou-se exame ginecológico dos animais para avaliação da saúde reprodutiva. A suplementação com colina protegida não alterou os níveis de beta-hidroxibutirato (BHBA), ácidos graxos não esterificados (AGNE), frutosamina, fator de crescimento semelhante a glicose I (IGF-I), status oxidante total (TOS), aspartato aminotransferase (AST) e gamaglutamiltransferase (GGT) no pós-parto. Não houve diferença também quanto à produção de leite. Aos 60 dias pós-parto, vacas suplementadas com colina protegida apresentaram menor número de casos de endometrite que vacas do grupo controle. A suplementação de colina protegida não alterou o perfil bioquímico e a produção de leite, mas reduziu o número de casos de endometrite no pós-parto de vacas leiteiras.(AU)

The study aimed to evaluate the effect of supplementation with protected choline on the energy profile, liver enzymes and reproduction in dairy cows in peripartum. Fifteen cows were divided into two groups: 8 received 80 grams of protected choline for 21 days pre-partum and 40 days postpartum, and 7 were considered control. Blood samples were collected on days 10, 20, 30, and 60 postpartum to evaluate the energy and hepatic profiles. After 60 days postpartum a gynecological examination of animals for evaluation of reproductive health was done. The supplementation with choline protected did not alter the beta-hydroxybutyrate levels (BHBA), non-esterified fatty acids (NEFA), fructosamine, like growth factor glucose (IGF-I), total oxidant status (TOS), aspartate aminotransferase (AST) and glutamiltrasferase range (GGT) postpartum. There was no difference in milk production. After 60 days postpartum, cows fed protected choline had fewer cases of endometritis that cows in the control group. The protected choline supplementation did not alter the biochemical profile and milk production, but reduced the number of endometritis cases in postpartum dairy cows.(AU)

Primed Low Frequency Transcranial Magnetic Stimulation Effects on Smoking Cue-Induced Craving

Abstract Repetitive Transcranial Magnetic Stimulation (rTMS) has shown the potential to reduce craving to smoke and the amount of cigarettes smoked. When low frequency rTMS is preceded by a priming session the cortical inhibitory effects are enhanced provoking a lasting performance. In this study we evaluate the brain regional activations and self-reported mood effect of one session of primed low frequency rTMS over the left dorsolateral prefrontal cortex (DLPFC) in response to smoking- related cues during temporary nicotine abstinence. Ten moderate nicotine dependent subjects participated in a crossover design, comparing a single session of active versus sham stimulation. After treatment, a functional Magnetic Resonance Imaging (fMRI) study was performed in all participants while completing a block-design cue reactivity paradigm. Our measures also included self-reported craving and mood score before and after every rTMS session. The number of daily smoked cigarette in a one week around period was also recorded. The results show that self-reported cravings were lower following the active stimulation; significant activation was found in areas associated with emotion control (anterior cingulate gyrus and subgenual area) and episodic memory (precuneus and posterior cingulate gyrus) as a result of the stimulation. No significant difference was found in the number of cigarettes smoked and mood scores between the sham and active condition during the one week period records. This study confirms the previous evidence that rTMS can reduce cigarette cravings and provides evidence of the neural mechanism of action that take place in order to achieve these neuromodulation effects.

Resumen La Estimulación Magnética Transcranea repetitiva (EMTr) ha demostrado ser capaz de reducir el deseo por fumar y el consume de cigarrillos. Cuando una baja frecuencia de EMTr es precedida por una sesión de primado, los efectos inhibidores de la corteza cerebral aumentan y también su duración. En este estudio se evalúan las activaciones regionales del cerebro y el efecto en el estado de ánimo auto-reportado de una sesión de EMTr de baja frecuencia primada sobre la corteza dorsolateral prefrontal izquierda en respuesta a estímulos visuales relacionados con el tabaco durante una abstinencia temporal a la nicotina. Diez sujetos con dependencia moderada a la nicotina participaron en un experimento de diseño cruzado, comparando una sola sesión de estimulación activa contra una simulada. Después del tratamiento, se llevó a cabo un estudio de Imagen por Resonancia Magnética funcional (IRMf) en todos los participantes mientras llevaban a cabo un paradigma visual de bloques. Las mediciones incluyeron una puntuación auto-reportada del deseo por fumar y estado de ánimo antes y después de cada sesión de EMTr. Los resultados muestran que el deseo por fumar era menor luego de una sesión activa de estimulación; se encontraron activaciones significativas en áreas asociadas con el control emocional (giro del cíngulo anterior) y memoria episódica (precúneo y giro del cíngulo posterior) como resultado de la estimulación. No se encontró diferencia significativa en el número de cigarros fumados entre la sesión activa y simulado durante el periodo evaluado. Tampoco se registraron diferencias en las evaluaciones de estado de ánimo luego del tratamiento. Este estudio confirma la evidencia previa de que la EMTr puede reducir el deseo por fumar y proporciona evidencia del mecanismo de acción neural que se lleva a cabo para lograr estos efectos neuromoduladores.

Aspectos nutricionales de la disfunción tiroidea en niños y adolescentes con síndrome de Down / Nutritional aspects of thyroid dysfunction in children and adolescents with down syndrome

La trisomía 21 o síndrome de Down (SD) es la causa más frecuente de retardo mental de origen cromosómico, asociada con disfunción tiroidea (DT) y obesidad como problema de salud pública. El hipotiroidismo es la forma más común de DT asociada con SD, con prevalencia cercana a 50%. Objetivo: Determinar la prevalencia de disfunción de tiroidea y aspectos relacionados con el estado nutricional de niños y adolescentes con SD. Métodos: Se realizó un estudio prospectivo y transversal, en 17 niños con SD (11,5 ± 3,0 años), seleccionados aleatoriamente. El estudio empleo evaluación antropométrica, entrevista dietética con recordatorio de 72 horas y determinación de hormonas tiroideas. El análisis estadístico requirió SPSS 17, con 95% IC, p <0,05. Resultados: Individuos con SD presentaron sobrepeso en 47,1% (n=8). Se observó hipotiroidismo subclínico en 71,4% (n=10), mientras que 28,6% (n=4) mostraron hipotiroidismo clínico. El consumo de yodo (53,4 ± 12,0 vs 130,5 ± 18,1; p<0,05), fenilalanina (0,1 ± 0,0 vs 2,3 ± 0,5; p<0,05) y tirosina (0,1 ± 0,0 vs 2,3 ± 0,5; p<0,05) fue bajo comparado con la media de requerimientos para estos nutrientes tiroideos en niños y adolescentes con SD. El consumo de yodo y concentraciones de T4 (r=0,512; p<0,03) mostraron correlación positiva; asimismo, el consumo de fenilalanina y tirosina (r=0,803; p<0,001). Conclusiones: Los trastornos de organificación de hormonas tiroideas en SD no justifican la DT. Se sugiere suplementación nutricional en etapas precoces de la vida, facilitando un estado eutiroideo que propicie el desarrollo físico y cognitivo en individuos con SD. Palabras clave: Síndrome de Down, disfunción tiroidea, nutrición, niños, adolescentes.

Trisomy 21 or Down Syndrome (DS) is the most common cause of mental retardation with chromosomal origin, associated with thyroid dysfunction (TD) and obesity as a public health problem. Hypothyroidism is the most common form of TD associated with DS, with nearly 50% prevalence. Objective: To determine the prevalence of thyroid dysfunction and aspects related to the nutritional status of children and adolescents with DS. Methods: A prospective and cross-sectional study in 17 children with DS (11.5 ± 3.0 years) were randomly selected. The study anthropometric, dietary interview with 72 hours and determination of thyroid hormones was realized. The required statistical analysis SPSS 17 with 95% CI, p <0.05. Results: Individuals with DS were overweight in 47.1% (n = 8). Subclinical hypothyroidism was observed in 71.4% (n = 10), while 28.6% (n = 4) showed clinical hypothyroidism. The consumption of iodine (53.4 ± 12.0 vs 130.5 ± 18.1; p <0.05), phenylalanine (0.1 ± 0, 0 vs 2.3 ± 0.5; p <0.05) and tyrosine (0.1 ± 0.0 vs 2.3 ± 0.5; p <0.05) was low compared to the average requirement for these thyroid nutrients in children and adolescents with DS. Consumption of iodine and T4 concentrations (r = 0.512; p <0.03) showed positive correlation; Also, consumption of phenylalanine and tyrosine (r = 0.803; p <0.001). Conclusions: Organification disorders of thyroid hormones in DS does not justify the TD. Nutritional supplementation of thyroid nutrients providing a euthyroid condition that improves physical and cognitive development in individuals with DS is suggested.

Evaluación del Estado Nutricional de los niños que participaron en el Programa “Activarse” durante el 2011 / Assessment of the Nutritional Status of Children Participating in the Program "Activarse" in 2011 / Avaliação do Estado Nutricional das crianças que participaram do Programa “Activarse” durante 2011

Introducción: Conocer el estado nutricional de los niños que participan del Programa Activarse es de suma importancia para evaluar y caracterizar a la población destinataria y poder optimizar las actividades y contenidos implementados. Material y Método: Estudio descriptivo y transversal. Se realizó una valoración antropométrica de 534 niños, de entre 5 y 10 años de edad, de 3 colegios ubicados en la Provincia de Buenos Aires que participan del programa Activarse desarrollado por Asociación Conciencia y Fundación PepsiCo. Se calculó la puntuación Z de los indicadores Talla/Edad y IMC/Edad. Para establecer el diagnóstico, se utilizó el software Anthro Plus1, al igual que los puntos de corte, salvo en el indicador Talla/Edad que se consideró como riesgo o alerta de baja talla a los niños que presentaban una puntuación Z entre -1,5 y 2. Resultados: La muestra total estuvo compuesta por un 55.4% (n=296) de niñas y un 44.6% (n=238) de niños. Elíndice Talla/Edad arrojó que el 6,5 % de la población tiene un diagnóstico de alerta o baja talla, distribuidos en forma pareja entre niños y niñas. A través del indicador IMC/Edad, se obtuvo que el 47,4% de los niños estudiados se encontraba en el binomio sobrepeso-obesidad. Los niños que presentaron exceso de peso, en su mayoría pertenecen al sexo masculino (52,5% vs 43,2% de sexo femenino). Conclusión: Se debe prestar especial atención al exceso de peso encontrado en la población evaluada, debiendo ser el foco prioritario en las intervenciones planificadas.

Introduction: Knowing the nutritional status of children participating in the program /"Activarse/" is extremely important in order to assess and characterise the target population and be able to optimize the activities and contents implemented. Materials and Methods: Cross-sectional descriptive study. An anthropometric assessment of 534 children aged 5-10 years, from 3 schools located in the province ofBuenos Aires which participate in the program"Activarse" developed by the Conciencia Association andthe PepsiCo Foundation. The z-score was calculated forthe height/age and BMI/age indicators. In order to establish the diagnosis, the Anthro Plus®1 software was used, as well as the cut-off points, but not in the height/age indicator wich was considered a sign of risk or alert of low height in children with a z-score between -1,5 and 2. Results: The overall sample was 55.4% (n=296) girls and 44.6% (n=238) boys. The height/age index showed that 6.5% of the population has an alert diagnosis or low height, distributed equally among boys and girls.Through the BMI/age indicator we could see that 47.4% of te evaluated children were overweight or obese; these were mostly boys (52,5% versus 43,2% girls). Conclusion: Special attention should be paid to the excessive weight found in the population evaluated, which must be a priority for future planned interventions.

Introdução: Conhecer o estado nutricional das crianças que participam do Programa “ActivaRSE” é fundamental para avaliar e caracterizar a população destinatária e poder otimizar as atividades e conteúdos implantados. Material e Método: Estudo descritivo e transversal. Foi realizada uma valorização antropométrica de 534 crianças entre 5 e 10 anos de idade, de 3 colégios localizados na Província de Buenos Aires que participam do programa “ActivaRSE” desenvolvido pela Associação Consciência e Fundação PepsiCo. Calculou-se a pontuação Z dos indicadores Estatura/Idade e IMC/Idade. Para estabelecer o diagnóstico, foi utilizado o software Anthro Plus, assim como os pontos de corte, salvo no indicador Estatura/Idade que foi considerado como risco ou alerta de baixa estatura em crianças que apresentavam uma pontuação Z -1,5 y 2. Resultados: A amostra total esteve composta por 55.4% (n=296) de meninas e 44.6% (n=238) de meninos. O índice Estatura/Idade indicou que 6,5% da população tem um diagnóstico de alerta ou baixa estatura, distribuídos de forma similar entre meninos e meninas. Através do indicador IMC/Idade, constatou-se que 47,4% das crianças estudadas encontravam-se no binômio sobre peso-obesidade. As crianças que apresentaram excesso de peso, na maioria pertenciam ao sexo masculino 52,5% vs 43,2% de sexo feminino). Conclusão: É preciso prestar uma atenção especial no excesso de peso encontrado na população avaliada, devendo ser o foco prioritário nas intervenções planejadas.

Initiation and s uppression of sperm motility is osmolality dependent in two South American fish species: streaked prochilod (Prochilodus lineatus) and piracanjuba(Brycon orbignyanus)

Solutions to induce or suppress the initiation of sperm motility in fish ha ve been used to improve reproductive success during artificial fertilization and preservation techniques . The aim of the present study was to evaluate the effects of three solutions (NaCl, glucose , and BTS™) - each prepared with 10 different osmolalities - on the initiation and suppression of fresh sperm motility in Prochilodus lineat us and Brycon orbignyanus . Sperm was diluted in each of the 30 solution s and immediately observed under a light microscope to determine which solution s trigger ed or suppress ed the initiation of sperm motility. When present, motility rate ( % motile sperm ) w as determined at 0, 30 , and 120 s post - activation and the motility quality score ranging from 0 ( no movement ) to 5 ( rapidly swimming sperm) was determined at 0 and 30 s post - activation . Osmolality , but not solution composition , significantly affected both motility rate and quality score . Solutions at osmolali ties up to 270 mOsm/kg in P. lineatus and up to 180 mOsm/kg in B. orbignyanus induced motility in at least 60% of sperm , with a minimum quality score of 3.0 , and were therefore classified as activating agents. The greatest motility at 0 , 30 , and 120 s post - activation was observed with solutions ranging from 135 to 225 mOsm/kg for P. lineatus and at 135 mOsm/kg for B. orbignyanus . On the other hand, solutions ranging from 360 to 450 mOsm/kg in P. lineatus and 270 to 450 mOsm/kg in B. orbignyanus suppressed motility in at least 95% of sperm and were classified as immobilizing media . The osmolality of the surrounding medium is the key factor in the initiation or suppression of sperm motility in P. lineatus a nd B. orbignyanu.

Lumbalgia mecánica crónica en pacientes con vértebras limbus anterior: revisión de la literatura y presentación de tres casos clínicos / Chronic mechanical low back pain in patients with anterior limbus vertebra: a review of the literature and a study of three cases

La vertebra limbus es un hallazgo radiológico de prevalencia desconocida y significado incierto. Se trata de un defecto óseo que afecta al margen de los cuerpos vertebrados, con un fragmento desprendido de morfología triangular y bordes escleróticos. Se produce como consecuencia de una herniación intraósea del núcleo pulposo a través del anillo apofisario vertebral, antes de la fusión completa del cuerpo vertebral. En este artículo se describe tres casos clínicos de pacientes jóvenes (de 19, 20 y 22 años), reclusos en el Centro Penitenciario de Jóvenes (Barcelona, España), diagnosticados de lumbalgía mecánica crónica, y portadores de vértebras limbus lumbar. Nuestros hallazgos, y la revisión sistemática de la escasa literatura disponible, ponen en duda algunas convicciones actuales sobre la vértebra limbus.

The limbus vertebra is a radiological finding of unknown prevalence and uncertain significance. This is a bone defect which affects themargin of the vertebral bodies, with a triangular-shaped detached fragment and sclerotic margins. This occurs as a result of an intraosseus herniation of the nucleus pulposus through the vertebral apophyseal ring before the complete fusion of the vertebral body. In this paper we describe the clinical cases of three young patients(aged 19, 20 and 22), in mates of the Young Offenders Institute of Barcelona, Spain, who were diagnosed with chronic mechanical lowback pain and suffered from lumbar limbus vertebrae. Our findings, and the systematic review of the limited literature available, question some of the current beliefs about limbus vertebrae.

Anemia y depleción de las reservas de hierro en adolescentes gestantes de una zona urbana y rural del estado Zulia, Venezuela / Anemia and iron depleted reserves in pregnant adolescents from urban and rural area, Zulia state, Venezuela

The objective of the present study was to analyze the prevalence of anemia and of iron deposits depletion (FeD) in pregnant adolescents from an urban (U) and rural (R) area. Methods: We studied 214 pregnant adolescents (U=100; R=114), from low socio-economic status. Clinical obstetric, nutritional situation and biochemical assessment was performed. Anemia in pregnant adolescents was determined during the first and third trimesters when Hh¡, 110g/L, second trimester Hb≤105g/L. FeD: ferritin <15µg/L; insufficient reserves of iron IRI=15-20µg/L and normal reserves of iron (RIN)>20µg/L, as recommended by WHO and the International Anemia Consultative Group. The statistical significance was considered when p<0.05. Results: The prevalence of anemia in pregnant adolescents with depleted reserves of iron (FeD+IRI) was 45.79% (R=22.90% vs U=22.89%); reserves of iron without anemia was 30.37%. (R=17.29% vs. U=13.08%); and anemia with RIN was 9.34% (R=6.07% vs. U=3.27%); there was a statistically significant association to urban (2=8.1282 p=0.0172) and rural area (2=11.6270p=0.0030). Conclusion: Iron deficiency anemia is a common complication in pregnant adolescents from rural and urban areas. This situation is associated with nutritional bad habits in adolescents. Strategies for recovery nutrition and education in primary health care and maternal health programs are needed.

Objetivo: Analizar la prevalencia de anemia y depleción de las reservas corporales de hierro (DRFe) en adolescentes gestantes urbanas (U) y rurales (R). Metodología: Fueron estudiadas 214 adolescentes gestantes (U=100 y R=114), de baja condición socio-económica. Se realizó una evaluación clínica-obstétrica, nutricional y bioquímica. Según recomendaciones de la OMS y el Grupo Consultivo Internacional de Anemia se consideró anemia en gestantes, I y III trimestre Hb≤110g/L, y II trimestre Hb≤105g/L. DRFe= ferritina<15ug/L; reservas insuficientes de hierro(RIFe)=ferritina 15-20ug/L y reservas de hierro normal (RFe normal)=ferritina >20ug/L. Se consideró significancia estadística p<0,05. Resultados: En las adolescentes gestantes la prevalencia de anemia con afectación de las reservas corporales de hierro (RIFe+DRFe) fue 45,79% (rurales=22,90% vs urbanas =22,89%), afectación de las reservas corporales de hierro (RIFe + DRFe) sin anemia 30,37% (R=17,29% vs. U=13,08%) y anemia con reservas corporales de hierro normal 9,34% (R=6,07% vs. U=3,27%); mostrando asociación estadísticamente significativa para la zona urbana (χ²=8,1282 p=0,0172) y la zona rural (χ²=11,6270 p=0,0030). Conclusiones: La anemia ferropénica es una complicación frecuente en adolescentes gestantes rurales y urbanas, asociada a malos hábitos alimentarios propios de la edad. Es necesario, incorporar estrategias de educación y recuperación nutricional en la atención primaria de salud y programas de salud maternoinfantil.

Procaína: Efeitos farmacológicos e toxicológicos / Procaine: Pharmacological and toxicological effects

Procaine is a local anesthetic used by dentists for decades. Nowadays it is being used to treat depression, increase the libido and act on inflammatory conditions and also to induce weight loss, among other uses. However, there has been criticism of such treatments with this substance, alone or in combination. The lack of a scientific basis makes its use subjective and unfounded and often potentially harmful to the individual. Therefore, the aim of this review is to find scientific evidence of systemic actions of procaine that demonstrate its efficacy for such purposes. From a review of the scientific literature, it was concluded that, except for a possible antidepressant effect, so far there are no data proving the alleged effects of procaine. In view of this, the current use of this substance in the treatment of chronic diseases or as an anti-aging drug would not be justified. Moreover, this review emphasizes the need for pharmacological and toxicological studies on procaine and the need to carry out in vivo and in vitro safety trials on pharmaceutical preparations containing this substance, in order to prove or disqualify the indications for its use.

A procaína é um anestésico local utilizado há décadas por dentistas. Atualmente, tem sido utilizada para tratar a depressão, aumentar a libido e agir em processos inflamatórios e no emagrecimento, entre outras utilidades. Porém, existem críticas acerca do tratamento com essa substância isolada ou associada. A falta de embasamento científico para sua utilização torna seu uso infundado e subjetivo, podendo ser muitas vezes nocivo ao indivíduo. Portanto, este artigo tem como objetivo buscar evidências científicas das ações sistêmicas da procaína que comprovem seus efeitos para tais finalidades. Foi realizado um levantamento na literatura científica e concluiu-se que, exceto por um possível efeito antidepressivo, até o momento não existem dados que comprovem os efeitos alegados para a procaína. Devido a isso, os usos atuais não se justificariam no tratamento de doenças crônicas ou no combate ao envelhecimento. Além disso, esta revisão enfatiza a necessidade da realização de estudos para avaliação farmacológica e toxicológica da procaína, bem como a necessidade de aplicar-se ensaios in vivo e in vitro na avaliação da segurança de preparações farmacêuticas que contenham essa substância, a fim de comprovar as inúmeras indicações de uso.