Curcumin alleviated oxidation stress injury by mediating osteopontin in nephrolithiasis rats

Purpose: To explore the role and mechanism of curcumin (Cur) in reducing oxidative stress damage in rats with nephrolithiasis induced by ethylene glycol (EG). Methods: Thirty male rats were divided into normal control, model, positive (10% potassium citrate), Cur-10 (10 mg/kg curcumin) and Cur-20 (20 mg/kg curcumin) groups. Results: The results of kidney tissue section stained by hematoxylin-eosin and von Kossa showed that curcumin treatment can inhibit the formation of kidney stones. The biochemical test results showed that the urea (Ur), creatinine (Cr), uric acid (UA), inorganic phosphorus and Ca2+ concentrations in urine decreased after being treated with curcumin. There were significant differences between different doses of curcumin (P < 0.05). Compared with the Cur-10 group, Cur-20 had a more significant inhibitory effect on malondialdehyde (MDA) (P < 0.05). In addition, reverse transcription polymerase chain reaction (PCR) detection and immunohistochemical results indicated that the osteopontin (OPN) in the kidney was significantly reduced after curcumin treatment. Conclusion: Curcumin could reduce the oxidative stress damage caused by EG-induced kidney stones.

Bond strength of experimental root canal sealers based on mta and butyl ethylene glycol disalicylate

Abstract This study investigated the bond strength of two experimental root canal sealers based on MTA and butyl ethylene glycol disalicylate: MTAe and MTAe-HA. The reference materials used for comparison were AH Plus and MTA Fillapex. Twenty human upper incisors were selected and one 1 mm slice was obtained from the cervical third of each root. On the coronal surface of each slice, four 0.9 mm wide holes were drilled through the dentine. Standardized irrigation was performed and holes were filled with one of the four tested sealers: AH Plus, MTA Fillapex, MTAe, and MTAe-HA. The filled slices were stored in a PBS solution (pH 7.2) for 7 days at 37 °C. A push-out assessment was performed with a 0.7 mm plunger tip. Load was applied at a crosshead speed of 0.5 mm/min until sealer displacement. The results were expressed in MPa. The Kruskal-Wallis test was applied to assess the effect of each sealer on the push-out bond strength. Mann-Whitney with Bonferroni correction was used to isolate the differences. The alpha-type error was set at 0.05. Significant differences among medians values obtained by materials were observed (p<0.001). AH Plus displayed the highest value of bond strength (p<0.001). In contrast, MTA Fillapex presented the lowest bond strength among all tested sealers (p<0.001). Experimental sealers showed intermediary bond strength values, with no statistical differences between them (p>0.05). In conclusion, experimental root canal sealers presented suitable bond strength outcomes when compared to MTA Fillapex.

Resumo Esse estudo investigou a resistência de união de dois cimentos endodônticos experimentais à base de MTA e butiletilenoglicol dissalicilato: MTAe e MTAe. Os materiais de referência utilizados para comparação foram os cimentos endodônticos MTA Fillapex e AH Plus. Vinte incisivos superiores humanos foram selecionados e um slice dentinário de 1 mm de espessura foi obtido do terço cervical de cada raiz. Na superfície coronária de cada slice, quatro orifícios com 0,9 mm de diâmetro foram confeccionados através da dentina. Uma irrigação padronizada foi realizada e os orifícios foram preenchidos com um dos quatro cimentos endodônticos avaliados: AH Plus, MTA Fillapex, MTAe, e MTAe-HA. Os slices preenchidos foram armazenados em solução PBS (pH 7,2) durante 7 dias a 37°C. O ensaio de push-out foi realizado por meio de um dispositivo com 0,7 mm de diâmetro. A carga foi aplicada com a velocidade de 0,5 mm/min até a obtenção de deslocamento do material obturador. Os resultados foram expressos em MPa. O teste de Kruskal-Wallis foi aplicado para avaliar o efeito da resistência de união de cada cimento endodôntico. O teste de Mann-Whitney com correção de Bonferroni foi utilizado para isolamento das diferenças. O erro do tipo-alfa foi fixado em 0,05. Diferenças significantes entre os valores de medianas obtidos pelos materiais foram observados (p<0,001). O AH Plus demonstrou os maiores valores de resistência de união (p<0,001). Em contraste, o MTA Fillapex apresentou a menor resistência de união entre todos os cimentos testados (p<0,001). Os cimentos experimentais demonstraram valores intermediários, com ausência de diferenças estatísticas entre si (p>0,05). Em conclusão, os cimentos endodônticos experimentais à base de MTA e butiletilenoglicol dissalicilato apresentaram resultados adequados de resistência de união quando comparados ao MTA Fillapex.

Combination of ethylene glycol with sucrose increases survival rate after vitrification of somatic tissue of collared peccaries (Pecari tajacu Linnaeus, 1758) / Combinação de etilenoglicol com sacarose aumenta a taxa de sobrevivência após a vitrificação de tecido somático de catetos (Pecari tajacu Linnaeus, 1758)

The cryopreservation of somatic tissue in collared peccaries promotes an alternative source of genetic material of this specie. The solid-surface vitrification (SSV) is a great option for tissue conservation; nevertheless, the optimization of SSV requirements is necessary, especially when referred to cryoprotectants that will compose the vitrification solution. Therefore, the aim was to evaluate the effect of the presence of 0.25 M sucrose in addition to different combinations (only or association) and concentrations (1.5 M or 3.0 M) of ethylene glycol (EG) and/or dimethyl sulfoxide (DMSO) in the somatic tissue vitrification of collared peccaries. Subsequently, we tested six combinations of cryoprotectants with or without sucrose in Dulbecco modified Eagle medium (DMEM) plus 10% fetal bovine serum (FBS). Thus, 3.0 M EG with sucrose was able to maintain normal tissue characteristics compared with non-vitrified (control), especially for the volumetric ratio of epidermis (61.2 vs. 58.7%) and dermis (34.5 vs. 36.6%), number of fibroblast (90.3 vs. 127.0), argyrophilic nucleolar organizer region (AgNOR) ratio (0.09 vs. 0.17%) and nucleus area (15.4 vs. 14.5 µm2) respectively. In conclusion, 3.0 M EG with 0.25 M sucrose and 10% FBS resulted in a better cryoprotectant composition in the SSV for somatic tissue of collared peccaries.(AU)

A criopreservação de tecido somático em catetos promove uma fonte alternativa de material genético nesta espécie. A vitrificação em superfície sólida (VSS) é uma ótima opção para a conservação do tecido; contudo, a otimização dos requerimentos da VSS é necessária, especialmente quanto aos crioprotetores que irão compor a solução de vitrificação. Portanto, o objetivo foi avaliar o efeito da presença de 0,25 M de sacarose em adição com diferentes combinações (individual ou associação) e concentrações (1,5 M ou 3,0 M) de etilenoglicol (EG) e/ou dimetilsulfóxido (DMSO) na vitrificação de tecido somático de catetos. Subsequentemente, nós testamos seis combinações de crioprotetores com ou sem sacarose em meio de Eagle modificado por Dulbecco (DMEM) acrescido de 10% de soro fetal bovino (SFB). Assim, 3,0 M de EG com sacarose foi capaz de manter as características normais do tecido comparado com o não vitrificado (controle), especialmente para a proporção volumétrica da epiderme (61,2 vs. 58,7%) e derme (34,5 vs. 36,6%), número de fibroblastos (90,3 vs. 127,0), razão da região argirófila organizadora de nucléolo (AgNOR) (0,09 vs. 0,17%) e área do núcleo (15,4vs.14,5 µm2), respectivamente. Em conclusão, 3,0 M de EG com 0,25 M de sacarose e 10% de SFB resultaram na melhor composição de crioprotetores na VSS para tecido somático de catetos.(AU)

Freezing goat embryos at different developmental stages and quality using ethylene glycol and a slow cooling rate / Congelação de embriões caprinos em diferentes estádios de desenvolvimento e qualidade utilizando etileno glicol e taxa de resfriamento lenta

The efficiency of an alternative freezing protocol for goat embryos of different morphology and quality was tested. Fifty-eight embryos on Day 6-7 stage were transferred as fresh or after freeze-thawing (n=29/group). For freezing, embryos were placed into 1.5M ethylene-glycol solution for 10min. During this time, they were loaded in the central part of 0.25mL straw, separated by air bubble from columns containing PBS/BSA 0.4% plus 20% BFS. Straws were then frozen using a freezing machine from 20ºC to -6ºC at a cooling rate of 3ºC/min, stabilization for 15min (seeding after 5min), from -6 C to -32ºC at 0.6 C/min,and plunged into liquid nitrogen. Frozen embryos were thawed for 30s at 37ºC in a water bath. Embryos subjected to fresh transfer were maintained in holding medium (37ºC). Fresh and frozen-thawed embryos were transferred at day 7 post-estrus to 30 recipients. Kidding and kid born rates were similar (P> 0.05), respectively, for recipients receiving fresh (66.7% or 10/15; 55.2% or 16/29) or frozen-thawed (60% or 9/15; 51.7% or 15/29) embryos. The cryopreservation of goat embryos using slow-freezing protocol and 1.5MEG resulted in similar efficiency rates of fresh embryos.(AU)

Este estudo testou a eficiência de protocolo alternativo de criopreservação de embriões caprinos de diferentes qualidades morfológicas. Foram utilizados 58 embriões, coletados entre o sexto e o sétimo dia do ciclo estral (n=29/grupo). Embriões congelados passaram por solução 1,5M etilenoglicol por 10min e foram aspirados durante esse tempo para parte central de palheta 0,25mL, separada por bolhas de ar de colunas contendo PBS 0,4% BSA e 20% SFB. As palhetas foram congeladas em máquina de congelação de 20ºC a -6ºC, com taxa de resfriamento de 3ºC/min, estabilização por 15min (seeding após 5min), -6ºC a -32ºC a 0,6ºC/min, e imersas em nitrogênio líquido. Os embriões foram descongelados por 30s a 37ºC, em água. Embriões frescos foram mantidos em solução de manutenção (37ºC). Embriões frescos e congelados/descongelados foram transferidos para 30 receptoras no sétimo dia do ciclo estral. A taxa de partos e a de crias nascidas (respectivamente) foram similares (P>0,05) para receptoras recebendo embriões frescos (66,7% ou 10/15; 55,2% ou 16/29) ou congelados/descongelados (60,0% ou 9/15; 51,7% ou 15/29). O protocolo de criopreservação de embriões utilizado no presente estudo resultou em índices de eficiência semelhantes aos de embriões frescos.(AU)

Tratamiento médico del "body packer" entre los años 2014 y 2016 en un hospital universitario del caribe colombiano / Medical management of the "body packer" at a university hospital in the colombian caribbean in the period 2014-2016

Introducción. El término "body packer" hace referencia a sujetos portadores de objetos intraabdominales extraños, que contienen drogas ilícitas con fines de contrabando. La mayoría son pacientes asintomáticos, en quienes se instaura conducta expectante, observación clínica estrecha y administración de medicamentos para la evacuación de los paquetes, con miras a prevenir posibles complicaciones, como obstrucción intestinal o intoxicación, asociadas a su transporte intraabdominal. Materiales y método. Se llevó a cabo un estudio transversal de linealidad retrospectiva en pacientes admitidos en la E.S.E Hospital Universitario del Caribe entre los años 2014 y 2016,bajo la sospecha diagnóstica de "body packer". Luego de una revisión de las bases de datos institucionales se analizaron variables demográficas y clínicas de los sujetos incluidos en el estudio. Resultados. Se incluyeron 4 pacientes de género masculino, entre los 22 y 66 años de edad. La cantidad de cápsulas transportadas en promedio fue de 43, para una máxima de 74. La cocaína fue la sustancia que más se identificó. Para la evacuación de los paquetes se empleó irrigación intestinal con polietilenglicol. El tiempo de evacuación máximo fue de 48 horas y no hubo complicaciones asociadas al manejo proporcionado. Discusión. Estudios respecto al tema, como este, confirman la seguridad del manejo conservador del paciente asintomático y apoyan el uso de polietilenglicol dada su efectividad para lograruna limpieza intestinal completa y por su bajo riesgo de complicaciones asociado a su uso en comparación con otros métodos, así como la menor necesidad de intervenciones quirúrgicas. Se requieren estudios prospectivos aleatorizados controlados a partir de los cuales se determinen, con base en mayor evidencia, las mejores prácticas a seguir

Introduction. The term "body packer" refers to subjects carrying intraabdominal foreign objects that contain illicit drugs for contraband purposes. The majority of patients are asymptomatic, in whom expectant management is established, with close clinical observation and administration of medications for evacuation of the packages, with prevention of possible complications such as intestinal obstruction or intoxication associated with intraabdominal transport. Materials and method. A retrospective linearity cross sectional study was carried out in patients admitted to the Hospital Universitario del Caribe, Cartagena, Colombia, under the diagnostic suspicion of "body packer" in the period 2014 and 2016. After a review of the institutional databases, demographic and clinical variables of the study subjects were analyzed. Results. Four patients were included, male, ages 22 to 66 years. The average number of capsules transported was 43, with maximum of 74. Cocaine was the substance mainly identified. Intestinal irrigation with polyethylene glycol was used for intestinal evacuation. The maximum evacuation time was 48 hours and there were no complication associated with the given management. Discussion. The existing studies on the subject, as well as this one, confirm the safety of the conservative management in the asymptomatic patient and support the effectiveness of polyethyleneglycol in achieving complete intestinal cleansing and the low risk of complications associated with its use with respect to other methods, together with diminished need for surgical intervention. Controlled randomized prospective studies are required to provide greater evidence in order to determine the best practice to be followed

Effect of mitochondrial potassium channel on the renal protection mediated by sodium thiosulfate against ethylene glycol induced nephrolithiasis in rat model

Purpose: Sodium thiosulfate (STS) is clinically reported to be a promising drug in preventing nephrolithiasis. However, its mechanism of action remains unclear. In the present study, we investigated the role of mitochondrial KATP channel in the renal protection mediated by STS. Materials and Methods: Nephrolithiasis was induced in Wistar rats by administrating 0.4% ethylene glycol (EG) along with 1% ammonium chloride for one week in drinking water followed by only 0.75% EG for two weeks. Treatment groups received STS, mitochondrial KATP channel opener and closer exclusively or in combination with STS for two weeks. Results: Animals treated with STS showed normal renal tissue architecture, supported by near normal serum creatinine, urea and ALP activity. Diazoxide (mitochondria KATP channel opening) treatment to the animal also showed normal renal tissue histology and improved serum chemistry. However, an opposite result was shown by glibenclamide (mitochondria KATP channel closer) treated rats. STS administered along with diazoxide negated the renal protection rendered by diazoxide alone, while it imparted protection to the glibenclamide treated rats, formulating a mitochondria modulated STS action. Conclusion: The present study confirmed that STS render renal protection not only through chelation and antioxidant effect but also by modulating the mitochondrial KATP channel for preventing urolithiasis.

Anti-inflammatory effects of royal jelly on ethylene glycol induced renal inflammation in rats

ABSTRACT Objective: In this study, anti-inflammatory effects of Royal Jelly were investigated by inducing renal inflammation in rats with the use of ethylene glycol. For this purpose, the calcium oxalate urolithiasis model was obtained by feeding rats with ethylene glycol in drinking water. Materials and Methods: The rats were divided in five study groups. The 1st group was determined as the control group. The rats in the 2nd group received ethylene glycol (1%) in drinking water. The rats in the 3rd group were daily fed with Royal Jelly by using oral gavage. The 4th group was determined as the preventive group and the rats were fed with ethylene glycol (1%) in drinking water while receiving Royal Jelly via oral gavage. The 5th group was determined as the therapeutic group and received ethylene glycol in drinking water during the first 2 weeks of the study and Royal Jelly via oral gavage during the last 2 weeks of the study. Results: At the end of the study, proinflammatory/anti-inflammatory cytokines, TNF-α, IL-1β and IL-18 levels in blood and renal tissue samples from the rats used in the application were measured. Conclusion: The results have shown that ethylene glycol does induce inflammation and renal damage. This can cause the formation of reactive oxygen species. Royal Jelly is also considered to have anti-inflammatory effects due to its possible antiradical and antioxidative effects. It can have positive effects on both the prevention of urolithiasis and possible inflammation during the existing urolithiasis and support the medical treatment.

Expression of vimentin and gfap protein of cerebral cortex and its impact on corticogenesis disorder as a result of 2-methoxyethanol / Expresión de vimentina y proteína gfap de la corteza cerebral y su impacto en el trastorno de corticogenesis como producto del 2-metoxietanol

One of the plastic base material, widely used in the plastics industry in various countries, is a ester phthalate. These compounds will be oxidizedin the body to 2-methoxyethanol (2-ME). Effect of 2-ME on human health and the environment depends on the number, duration and frequency of exposure. 2-ME and its metabolites in the body can damage cells and tissues. The body can be exposed by 2-ME through the air, water and soil. Western blot results showed that the protein Vimentin was detectable in the control group at GD-11 to 17, meanwhile GFAP protein was detachable in the control group atGD- 12 to GD-18. After administration 2-ME, the expression of Vimentinprotein were changed, and started at GD- 12 up to GD-18. whereas the expression of GFAP protein began at GD-11 up to GD-17. The Changes on timetable protein expression of Vimentin and GFAP affect corticogenesis disorder. The disorder caused by the existence of these proteins as a result of 2-Methoxyethanol. Disorder of corticogenesis process were sub-plate and cortical plate of the cerebral cortex of fetus brains of mice at GD-18. Generally, it can be concluded that changes inprotein expression of Vimentin and GFAP causedby 2-ME. The Vimentin more important during the period of fetal brain development. GFAP and Vimentin is a protein involved in response to damage caused by a teratogenic agent, so that cells in the cerebral cortex, has dedifferentiation.

Uno de los materiales a base de plástico, ampliamente utilizado en la industria en varios países, es un éster de ftalato. Estos compuestos se oxidan en el cuerpo a 2-metoxietanol (2-ME). El efecto del 2-ME en la salud humana y el medio ambiente depende de la cantidad, duración y frecuencia de exposición. El 2-ME y sus metabolitos en el cuerpo puede dañar las células y tejidos. El cuerpo puede ser expuesto al 2-ME a través del aire, agua y suelo. Los resultados de Western blot mostraron que la proteína vimentina fue detectable en el grupo de control en GD-11 a 17, por su parte proteína GFAP fue detectable en el grupo de control en GD-12 a GD-18. Después de la administración de 2-ME, la expresión de la proteína vimentina cambió, y comenzó a detectarse en GD-12 hasta GD-18, mientras que la expresión de la proteína GFAP se inició en GD-11 hasta GD-17. Los cambios en el momento de expresión de las proteínas vimentina y GFAP afectan produciendo trastornos de la corticogénesis. El trastorno causado por la existencia de estas proteínas como resultado de 2-metoxietanol a nivel del proceso corticogénesis fue en la subplaca y la placa cortical de la corteza cerebral del cerebro de fetos de ratones en GD-18. En general, se puede concluir que existen cambios en la expresión de las proteínas vimentina y GFAP causados por el 2-ME. La vimentina es muy importante durante el período de desarrollo del cerebro fetal. GFAP y vimentina son proteínas implicadas en la respuesta a los daños causados por un agente teratogénico, de modo que las células en la corteza cerebral presentan desdiferenciación.

Anti-Urolithiatic Effect of Cow Urine Ark on Ethylene Glycol-Induced Renal Calculi

Purpose To investigate the anti-urolithiatic effect of cow urine ark (medicinal distilled cow urine) on ethylene glycol (EG) induced renal calculi. Materials and Methods 36 male Wistar rats were randomly divided into 6 equal groups. Group I animals served as vehicle control and received distilled water for 28 days. Group II to VI animals received 1% v/v EG in distilled water for 28 days. Group II served as EG control. Group III and IV (preventive groups) received cow urine ark orally for 28 days in doses of 1 mL/kg and 2 mL/kg, respectively. Group V and VI (treatment groups) received 1 mL/kg and 2 mL/kg cow urine ark orally, respectively from 15th to 28th days. 24-hour urine samples were collected on day 0 and 28. Urine volume and oxalate levels were measured. On day 28, blood was collected for biochemical parameters. Animals were sacrificed and kidneys were harvested, weighed and histopathologically evaluated for calcium oxalate (CaOx) crystals. To calculate the percentage of inhibition of mineralization, simultaneous flow static in-vitro model was used. Results EG significantly increased urine oxalate, serum creatinine, blood urea level; kidney weight and CaOx deposits. Provision of cow urine ark resulted in significantly lower levels of urine oxalate, serum creatinine, blood urea and CaOx depositions as compared to Group II. (p value < 0.05) It also significantly restored kidney weight. (p value < 0.05) Cow urine ark inhibited 40% and 35% crystallization of CaOx and calcium phosphate, respectively. Conclusion Cow urine ark is effective in prevention and treatment of EG induced urolithiasis in Wistar rats. .

Prophylactic effect of coconut water (Cocos nucifera L.) on ethylene glycol induced nephrocalcinosis in male wistar rat

Purpose Many medicinal plants have been employed during ages to treat urinary stones though the rationale behind their use is not well established. Thus, the present study was proposed to evaluate the effect of coconut water as a prophylactic agent in experimentally induced nephrolithiasis in a rat model. Materials and Methods The male Wistar rats were divided randomly into three groups. Animals of group I (control) were fed standard rat diet. In group II, the animals were administrated 0.75% ethylene glycol in drinking water for the induction of nephrolithiasis. Group III animals were administrated coconut water in addition to ethylene glycol. All the treatments were continued for a total duration of seven weeks. Results and Conclusion Treatment with coconut water inhibited crystal deposition in renal tissue as well as reduced the number of crystals in urine. Furthermore, coconut water also protected against impaired renal function and development of oxidative stress in the kidneys. The results indicate that coconut water could be a potential candidate for phytotherapy against urolithiasis. .

Synthesis and MALDI-ToF characterization of dendronized poly(ethylene glycol)s

Well-defined hybrids of linear poly(ethylene glycol)s (PEGs) and dendritic polyesters were prepared via the dendronization of the alcohol end groups of the mono and difunctional linear PEGs. Though useful for rudimentary product characterization, GPC and NMR could not verify the overall structural purity of these linear-dendritic hybrids. On the other hand, the detailed data provided by MALDI-ToF mass spectrometry enabled confirmation of the high structural purity of the dendronized PEGs at each step of the dendronization procedure. The well-defined number of functionalities on these dendronized PEGs, renders them particularly useful for research in the biomedical sphere where functionality and purity are of the utmost importance. The MALDI-ToF mass spectrometric approach described herein represents a valuable technique for detailed monitoring of these dendronization reactions, as well as a variety of other polymer end group modifications.

Híbridos bem definidos de poli(etilenoglicol) lineares (PEGs) e poliésteres dendriméricos foram preparados via "dendronização" de álcool e grupos de PEGs lineares mono e bifuncionais. Embora úteis para a caracterização rudimentar de produtos, Cromatografia por Permeação em Gel e RMN podem não demonstrar a pureza estrutural global desses híbridos lineares dendríticos. Por outro lado, informações detalhadas provenientes de espectrometria de massas MALDI-ToF permitiram a confirmação de elevada pureza estrutural de PEGs "dendronizados" em cada passo do processo de "dendronização". O número de funcionalidades bem definidas destes PEGs "dendronizados", torna-os particularmente úteis para pesquisa na área biomédica, na qual funcionalidade e pureza são de grande importância. A abordagem de espectrometria de massas MALDI-ToF descrita aqui representa uma técnica valiosa para o monitoramento detalhado destas reações de "dendronização", bem como diversas modificações de outros polímeros e grupos.

Aerobic biodegradation of nonylphenol ethoxylates in shaking-flask test

Nonylphenol ethoxylates (NPEOs), which are widely used for industrial and domestic purposes, exert adverse effects on wildlife after being used and discharged into the environment. However, their ultimate biodegradability and biodegradation pathway remains unclear. In this study, the aerobic degradability of nonylphenol ethoxylates (NPEOs) by the acclimated microorganisms in active sludge was examined using shaking-flask tests. The degradation of benzene rings in NPEOs was determined using UV spectroscopy and high performance liquid chromatography (HPLC). Results showed that more than 80 percent of benzene rings were removed after 8-10 days of degradation, and the majority of NPEOs were also removed after 9 days of degradation, indicating NPEOs and the benzene rings could be ultimately degraded by microorganisms in acclimated active sludge. Electrospray ionization-mass spectroscopy (ESI-MS) analysis of biodegradation intermediates indicate that stepwise omega, beta-oxidation of EO chains or fission of EO chains, and further omega, beta-oxidation of alkyl-chain for short-EO-chain NPEOs constitute the main pathway in the early stage, and complete biodegradation occur when the benzene rings in these molecules are opened in the later stage.

Teratogenic effect of ethylene glycol-methyl cellosolve mixture in rats: I: reproductive damage / Teratogénesis por etilenglicol-metilcelosolve en ratas: I: daño reproductivo

We investigated the reproductive damage and teratogenic effect of an ethylene glycol-methyl cellosolve mixture on gestating Wistar rats, which received a daily intraperitoneal dose of different concentration of the mixture on day 8 of gestation until day 20. In rats treated with the mixture the number of live fetuses decreased and reabsorptions increased with increasing concentrations of the mixture, as well as the number of abnormal fetuses. We conclude that the ethylene glycol-methyl cellosolve mixture possesses a higher teratogenic potential than each of its constituents separately, producing reproductive damage, external fetal abnormalities, growth delay, and increased fetal death.

Se investigó el daño reproductivo y efecto teratogénico de una mezcla de etilenglicol y metilcelosolve en ratas gestantes, las cuales recibieron por vía intraperitoneal una dosis diaria, a diferentes concentraciones de la mezcla, del día 8 al día 20 de gestación. En las ratas tratadas con la mezcla el número de fetos vivos disminuyó y las reabsorciones y el número de fetos anormales aumentaron a mayor concentración de los solventes. Concluimos que la mezcla de etilenglicol-metilcelosolve tiene mayor efecto teratogénico que cuando actúan cada uno de los solventes por separado, produciendo daño reproductivo, anormalidades fetales externas, retraso del crecimiento y aumento de muerte fetal.

Teratogenic effect of ethylene glycol-methyl cellosolve mixture in rats: II: craniofacial and limb abnormalities / Teratogénesis por etilenglicol-metilcelosolve en ratas: II: anormalidades craneofaciales y de miembros

We investigated the teratogenic effect of an ethylene glycol-methyl cellosolve mixture on gestating Wistar rats, that received a daily intraperitoneal dose of different concentration of the mixture on day 8 of gestation until day 20. Multivariate analysis and Post-Hoc Bonferroni tests were used and relative risk and attributable fraction were calculated. In rats treated with the mixture the number of live fetuses decreased and reabsorptions increased with increasing concentrations of the mixture, as well as the number of abnormal fetuses. Abnormalities consisted mainly in atypical craniofacial morphology, protruding tongue, edema, signs of growth delay and shorter limbs, their frequency and severity increased at higher concentrations of the mixture. We conclude that the ethylene glycol-methyl cellosolve mixture possesses a higher teratogenic potential than each of its constituents separately, producing external fetal abnormalities, growth delay, and increased fetal death.

Se investigó el efecto teratogénicos de una mezcla de etilenglicol y metilcelosolve en ratas gestantes, las cuales recibieron por vía intraperitoneal una dosis diaria, a diferentes concentraciones de la mezcla, del día 8 al día 20 de gestación. Se utilizaron las pruebas de análisis multivariado y Post-Hoc de Bonferroni, y se calcularon el riesgo relativo y la fracción atribuible. En las ratas tratadas con la mezcla el número de fetos vivos disminuyó y las reabsorciones y el número de fetos anormales aumentaron a mayor concentración de los solventes. Las anormalidades fetales consistieron principalmente en morfología atípica craneofacial, protrusión de la lengua, edema, signos de retraso de crecimiento y acortamiento de extremidades, y su frecuencia y severidad se incrementó a mayor concentración de la mezcla. Concluimos que la mezcla de etilenglicol-metilcelosolve tiene mayor efecto teratogénico que cuando actúan cada uno de los solventes por separado, produciendo anormalidades fetales externas, retraso del crecimiento y aumento de muerte fetal.

Intoxicación con etilenglicol. Presentación de casos y revisión de la literatura / Intoxication with ethylene glycol. Presentation of cases and review of literature

El etilénglicol es un producto utilizado en la industria química. La ingesta o aspiración de esta sustancia es una emergencia médica que se debe diagnos� ticar y tratar de inmediato. Inicialmente produce un cuadro conocido como "embriaguez sin aliento alcohólico", seguido de toxicidad cardiopulmonar y renal con grave acidosis metabólica con brecha aniónica aumentada. En la mayoría de los Centros, no es posible determinar la concentración de eti� lénglicol en sangre, por lo que el diagnóstico inicial se basa en la anamnesis y en la presencia de acidosis metabólica grave con brecha aniónica elevada. El tratamiento consiste en soporte vital, adecuada infusión de fluidos y bicar� bonato de sodio, administración de etanol o fomepizol para antagonizar la enzima alcohol deshidrogenasa y, en algunos casos, hemodiálisis.(AU)

The ethylene glycol is a product used in the chemical industry. The intake or inhalation of this substance is a medical emergency that should be diagnosed and treated early. Initially it causes a condition known as "drunkenness with� out alcoholic breath", followed by cardiopulmonary and renal dysfunctions with severe metabolic acidosis and increased anion gap. Determination of blood levels is not available in most health care centers, so initial diagnosis should be based on history and the presence of metabolic acidosis with el� evated anion gap. Treatment consists of vital support, adequate fluid and bicarbonate infusion, administration of ethanol o fomepizole to antagonize the � genase and, in some cases, hemodialysis.(AU)

Efeito do glicerol, etilenoglicol, acetamida e leite desnatado na criopreservação de espermatozóides eqüinos / Effects of glycerol, ethylene glycol, acetamide, and dried skim milk in cryopreservation of equine sperm

Avaliou-se a eficácia de diferentes associações de crioprotetores no congelamento de sêmen eqüino. Foram utilizados três ejaculados de oito garanhões para testar o diluidor lactose-EDTA-gema de ovo com as seguintes associações de macromoléculas e crioprotetores: T1 - glicerol 5 por cento (controle); T2 - metilcelulose 0,5 por cento, rafinose 0,15g e acetamida 2,5 por cento; T3 - metilcelulose 0,5 por cento, rafinose 0,15g e acetamida 3,5 por cento; T4 - metilcelulose 0,5 por cento, rafinose 0,15g e acetamida 5 por cento; T5 - glicerol 5 por cento e 2,4g de leite desnatado; T6 - 1 por cento de glicerol, 4 por cento de etilenoglicol e 2,4g de leite desnatado; T7 - 5 por cento de etilenoglicol e 2,4g de leite desnatado. O sêmen foi diluído em meio Kenney (1:1), centrifugado a 400 x g por 12 minutos, ressuspendido nos diluidores para atingir a concentração de 100X10(6)/ml, envasado em palhetas de 0,5ml e congelado 3cm acima do nível de nitrogênio líquido por 10 minutos. O descongelamento foi realizado em banho-maria a 75ºC por sete segundos. Após o descongelamento foram avaliados: motilidade total e progressiva, vigor, morfologia espermática e integridade estrutural e funcional da membrana plasmática. O T1 apresentou os maiores valores de motilidade total e progressiva (38,4 por cento e 33,8 por cento, respectivamente). O vigor e os resultados do teste HO não diferiram entre os tratamentos. Os diluidores contendo leite em sua composição (T5, T6 e T7) apresentaram maiores valores de integridade funcional e estrutural da membrana plasmática. Pode-se concluir que as modificações incorporadas aos meios diluidores testados não resultaram em melhor efeito crioprotetor que o meio à base de glicerol 5 por cento no congelamento do sêmen eqüino.

The efficacy of different combinations of cryoprotectants for equine semen was evaluated. Three ejaculates of eight stallions were used to test the lactose-EDTA-egg yolk extender with the following association of cryoprotectants: T1 - glycerol 5 percent (control group); T2 - methyl cellulose 0.5 percent, raffinose 0.15g, and acetamide 2.5 percent; T3 - methyl cellulose 0.5 percent, raffinose 0.15g, and acetamide 3.5 percent; T4 - methyl cellulose 0.5 percent, raffinose 0.15g, and acetamide 5 percent; T5 - glycerol 5 percent and 2.4g of dried skim milk; T6 - glycerol 1 percent, ethylene glycol 4 percent, and 2.4g of dried skim milk; T7 - ethylene glycol 5 percent and 2.4g of dried skim milk. After collection, Kenney extender was added to the semen 1:1, and centrifuged at 400 x g for 12 minutes. Sperm pellets were diluted to reach 100x10(6) cells/ml. Spermatozoa were frozen in 0.5ml straws 3cm above the nitrogen level, during 10 minutes. Thawing of samples was done at 75ºC for seven seconds followed by immersion of the straw in a water bath at 37ºC for 30 seconds. Post-thaw total and progressive motilities and sperm vigor were evaluated. Sperm membrane integrities of the tail and caput, respectively, were evaluated by the hypoosmotic swelling test and fluorescent dyes. T1 showed the highest post-thaw total and progressive motilities (38.4 percent and 33.8 percent, respectively). No significant difference was found among treatments for vigor and hypoosmotic swelling test. T5, T6, and T7 showed higher post-thaw values for sperm membrane integrity. It may be concluded that the association of cryoprotectants used in this experiment did not result in better cryoprotectant effect than 5 percent glycerol for equine semen cryopreservation.

Efecto de la asociación L-glutamina - Etilenglicol en la criopreservación de semen equino / Effect of the association l-glutamine - ethylene glycol in equine semen cryopreservation

Con el objeto de mejorar la eficiencia en la criopreservación de semen equino, se evaluó el efecto de la asociación L-glutamina con el Etilenglicol y glicerol en el medio de congelación seminal. Se usaron 4 reproductores criollos Colombianos para completar un total de 21 muestras que fueron congeladas en diferentes medios de congelación constituidos de medio INRA 97 y crioprotectante según estudio: L-glutamine 80mM + Etilenglicol 2,5 por ciento (protocolo 1), L-glutamine 80mM + Glycerol 2,5 por ciento (protocolo 2), Etilenglicol 2,5 por ciento (rotocolo 3) y glycerol 2,5 por ciento (protocolo 4). la metodología de congelación fue: 60 minutos para descender la temperatura de 38ºC a 5ºC (0,55ºC/min) durante el transporte. Se centrifugaron las muestras a 600G/10min, se duluyó el semen con los cuatro protocolos en pajillas de 0,5 ml. Luego 60 minutos de equilibrio en refrigeración; 20 minutos en vapores de nitrógeno líquido y posterior inmersión. En la evaluación de la motilidad progresiva no se encontró diferencia significativa entre protocolos al tiempo 0 (P<0.6383), al tiempo 30 min (P<0.511) y al tiempo 60 min (P<0.1659)...

Partial purification of glucose-6-phosphate dehydrogenase by aqueous two-phase poly(ethyleneglycol)/phosphate systems

Glucose-6-phosphate dehydrogenase (G6PDH) is an important enzyme used in biochemical and medical studies and in several analytical methods that have industrial and commercial application. This work evaluated the extraction of G6PDH in aqueous two-phase system (ATPS) of poly(ethyleneglycol) (PEG)/phosphate buffer, using as enzyme source a medium prepared through commercial baker's yeast disruption. Firstly, the effects of PEG molar mass on the enzyme partition and of homogenization and rest on the system equilibrium were investigated. Afterwards, several ATPS were prepared using statistical analysis (2² factorial design). The results, including kinetic and thermodynamic parameters for the G6PDH activity, showed partial purification of this enzyme in ATPS composed of 17.5 percent (w/w) PEG400 and 15.0 percent (w/w) phosphate. A high enzymatic recovery value (97.7 percent), a high partition coefficient (351), and an acceptable purification factor (2.28 times higher than in cell homogenate) were attained from the top phase. So, it was possible to attain an effective enzyme pre-purification by separating some contaminants with a simple method such as liquid-liquid extraction in aqueous two-phase systems (ATPS).

Glicose-6-fosfato desidrogenase (G6PDH) é uma importante enzima usada em estudos bioquímicos e médicos, bem como em diversos métodos analíticos com aplicação comercial e industrial. Neste trabalho foi avaliado a extração da G6PDH em sistemas de duas fases aquosas (ATPS) constituídos por poli(etilenoglicol) (PEG)/tampão fosfato, usando como fonte de enzima um meio preparado por rompimento de leveduras de panificação comercial. Inicialmente foram investigados os efeitos da massa molar do PEG na partição da enzima e da homogeneização e repouso no equilíbrio do sistema. Na sequência, diversos ATPS foram preparados usando análise estatística (planejamento fatorial 2²). Os resultados, incluindo parâmetros cinéticos e termodinâmicos para a atividade da G6PDH, indicaram parcial purificação desta enzima em ATPS constituídos por 17,5 por cento (p/p) PEG400 e 15,0 por cento (p/p) fosfato. Um alto valor de recuperação enzimática (97,7 por cento), um alto coeficiente de partição (351), e um fator de purificação aceitável (2,28 vezes maior que em homogenato celular) foram obtidos na fase superior do sistema. Assim, foi possível alcançar uma pré-purificação eficaz da enzima separando alguns contaminadores aplicando um método simples tal como a extração líquido-líquido em sistemas bifásicos (ATPS).

Effect of vitrification on viability and chromosome abnormalities in 8-cell mouse embryos at various storage durations

This study was desµgned to investµgate the effect of vitrification and post-thaw survival and chromosomal aberrations caused by vitrification of vitrified 8-cell mouse embryos in comparison with a controligroup. To this purpose the survival rate and the frequency of chromosomal aberrations were assessed in frozen-thawed 8-cell mouse embryos after various storage durations in the presence of ethyleneiglycol as cryoprotectant. eight-cell mouse embryos were obtained from NMRI mice 3 days after mating. Retrieved embryos were transferred to vitrification solution containing ethyleneiglycol as cryoprotectant, then transferred into a vitrification straw using standard technique, and vitrified in liquid nitrogen. Sixigroups of embryos according to storage duration (24 hours, 1 and 2 weeks, 1-6 months) were frozen. After appropriate storage periods embryos were thawed and studied for their viability 4-6 hours after thawing and intact embryos were transferred to fresh medium containing colcemid. After 48 hours, the embryos were fixed and studied for their chromosome abnormalities using Tarkowsky's drying technique. Results indicate that freezing affects the viability and chromosome structure of embryos when compared with the controligroup. Furthermore increasing the storage duration reduces the viability and increases the chromosome aberrations of embryos (such as aneuploidy and polyploidy). This result mµght indicate that the effects of vitrification on the cytoskeleton or other cellular organelle mµght produce chromosomal alterations leading to cell death.