Tartrazina induce genotoxicidad en linfocitos de BALB/c Mus musculus / Tartrazine induces genotoxicity in BALB/c Mus musculus lymphocytes

RESUMEN Objetivos. Determinar el efecto genotóxico de la tartrazina en linfocitos de sangre periférica de Mus musculus BALB/c. Materiales y métodos. Se realizó un estudio experimental, a través de cinco grupos, con cinco ratones en cada uno. Se les registró el peso durante 17 semanas y, en la semana 15 se les administró suero fisiológico (control negativo), dicromato de potasio 25 mg/kg de peso corporal (pc) (control positivo) y tartrazina a dosis de 0,75 mg/kg pc, 7,5 mg/kg pc y 75 mg/kg pc, durante siete días, a excepción del control positivo que fue en dosis única. Luego, cada 24 h se obtuvo una muestra de sangre periférica de la cola y se realizó el frotis, secado y coloración. Posteriormente, se realizó el conteo de 1000 linfocitos por muestra de cada ratón, en todos los tratamientos. Resultados. Los tres tratamientos con tartrazina no causaron diferencias significativas en el peso de ratones a la semana 15, pero sí produjeron diferencias significativas en la frecuencia de linfocitos micronucleados, siendo el tratamiento con tartrazina de 75 mg/kg pc el de mayor efecto genotóxico, induciendo un promedio de 1,63 ± 0,08 linfocitos micronucleados, comparado con el control positivo que generó un promedio de 1,42 ± 0,08 linfocitos micronucleados. Conclusiones. La tartrazina produjo un efecto genotóxico, incrementando el número de linfocitos micronucleados, a dosis de 0,75; 7,5 y 75 mg/kg pc y no afecta el peso corporal durante siete días de administración en M. musculus BALB/c.

ABSTRACT Objectives. To determine the genotoxic effect of tartrazine on peripheral blood lymphocytes of BALB/c Mus musculus. Materials and methods. An experimental study was carried out using five groups, with five mice in each group. Their weight was registered for 17 weeks, and at week 15 they were administered physiological saline solution (negative control), potassium dichromate at 25 mg/kg body weight (bw) (positive control) and tartrazine at doses of 0.75 mg/kg bw, 7.5 mg/kg bw and 75 mg/kg bw, for seven days, with the exception of the positive control which was a single dose. Then, every 24 hours, a peripheral blood sample was obtained from the tail, which was then smeared, dried and stained. Subsequently, 1000 lymphocytes were counted for each sample from each mouse, for all treatment groups. Results. The three tartrazine treatments did not cause significant differences in the weight of mice at week 15, but did produce significant differences in the frequency of micronucleated lymphocytes, with the 75 mg/kg bw tartrazine treatment having the greatest genotoxic effect, inducing an average of 1.63 ± 0.08 micronucleated lymphocytes, compared to the positive control which obtained an average of 1.42 ± 0.08 micronucleated lymphocytes. Conclusions. Tartrazine produced a genotoxic effect, increasing the number of micronucleated lymphocytes, at doses of 0.75; 7.5 and 75 mg/kg bw and did not affect body weight during seven days of administration to BALB/c M. musculus.

Phytochemical composition, antifungal activity, in vitro and in vivo toxicity of Syzygium cumini (L) Skeels leaves extract / Composición fitoquímica, actividad antifúngica, toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (L) Skeels

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125µg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivotoxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

Este estudio determinó la composición fitoquímica, la actividad antifúngica y la toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (Sc). Así, se caracterizó mediante cromatografía de gases acoplada a espectrometría de masas y se sometió a determinación de Concentraciones Mínimas Inhibitorias (CMI) y Fungicidas (MFC) sobre cepas de referencia y clínicas de Candida spp. y mediante ensayos de cinética de crecimiento. La toxicidad se verificó mediante ensayos in vitro de hemólisis, fragilidad osmótica, actividad oxidante y antioxidante en eritrocitos humanos y por toxicidad sistémica aguda in vivo en larvas de Galleria mellonella. Se identificaron catorce compuestos diferentes en Sc, que mostraron actividad antifúngica (CMI entre 31.25-125 µg/mL) con efecto fungistático sobre Candida. En concentraciones antifúngicas, demostró baja citotoxicidad, actividad antioxidante y toxicidad in vivo insignificante. Por lo tanto, Sc demostró un potencial antifúngico prometedor, con baja toxicidad, lo que indica que este extracto puede ser un agente antifúngico alternativo seguro y eficaz.

Ensayo de inmunogenicidad y toxicidad aguda del candidato vacunal contra el cólera vax-COLER® / Immunogenicity and acute toxicity assay of the vaccine candidate against cholera vax-COLER®

Para el control y prevención del cólera humano se han llevado a cabo diferentes estrategias, entre las cuales, la vacunación es una de las medidas más eficaces. La evaluación preclínica de candidatos vacunales requiere de la demostración de la seguridad de los mismos, para lo cual los estudios toxicológicos son determinantes, al ser obligatorios y altamente regulados. Este estudio tuvo como objetivo demostrar la relevancia de las ratas Sprague Dawley como biomodelo a través de su respuesta inmunológica al candidato vacunal contra el cólera, vax-COLER®, utilizando la técnica de determinación de anticuerpos vibriocidas. Además, evaluar los efectos toxicológicos locales y sistémicos por la administración de una dosis de vax-COLER® a través de la evaluación de síntomas, del consumo de agua y alimentos, el peso corporal y estudios anatomopatológicos. La vacuna vax-COLER® resultó inmunogénica y no evidenció síntomas ni muertes, no hubo cambios en el peso corporal y los consumos de agua y alimentos se comportaron de forma similar entre todos los grupos. Los estudios anatomopatológicos solo mostraron cambios a nivel histológico en los ganglios linfáticos mesentéricos y placas de Peyer de los animales vacunados, con presencia de hiperplasia de los folículos secundarios subcapsulares, hallazgo que difirió significativamente con el resto de los grupos. Se concluye que la vacuna vax-COLER® es inmunogénica en ratas Sprague Dawley, demostrando la relevancia del biomodelo para la evaluación de la seguridad preclínica y que la aplicación de una dosis no produjo efectos tóxicos agudos generales ni locales(AU)

Different strategies have been carried out for the control and prevention of human cholera. Vaccination is one of the most effective strategies. Preclinical evaluation of vaccines needs to prove their safety; whereby toxicological studies are decisive. They are mandatory and highly regulated. This study was aimed to demonstrate the relevance of Sprague Dawley rats as a biomodel, through the immunological response to vax-COLER® cholera vaccine, using the technique of determination of vibriocidal antibodies. In addition, local and systemic toxicological effects were evaluated after administration of a dose of vax-COLER®; through the evaluation of symptoms, water and food consumption, body weight and anatomopathological studies. The vax-COLER® vaccine was immunogenic and showed no symptoms or deaths. No changes in body weight were detected, and food and water consumption were similar among all groups. The anatomopathological studies showed histological changes in the mesenteric lymph nodes and Peyer's patches of the vaccinated animals, with hyperplasia of the subcapsular secondary follicles, finding that differed significantly from the rest of the groups. It is concluded that vax-COLER® vaccine is immunogenic in Sprague-Dawley rats, demonstrating the relevance of the biomodel for the evaluation of preclinical safety, as well as that the application of a single dose did not produce acute general or local toxic effects(AU)

Efeito de análogos de curcumina, irradiados ou não por LED, como agentes antimicrobianos e indutores de proliferação e migração celular / Effect of curcumin analogues, irradiated or not by LED, as antimicrobial agents and inducers of cell proliferation and migration

O tratamento endodôntico de dentes permanentes jovens com infecções pulpares/periapicais antes de completar a rizogênese ainda é um desafio para a Endodontia e a Odontopediatria. Relatos científicos têm mostrado que a curcumina (CUR), um fitoquímico polifenólico, apresenta diversas propriedades terapêuticas, entre as quais, amplo espectro de ação antimicrobiana e a capacidade de induzir a proliferação e migração celular. Além disso, devido à sua capacidade excitatória na presença de luz, a CUR também tem sido utilizada como fotossensibilizante em terapia fotodinâmica associada ao LED (light emitting diode), promovendo aumento dos seus efeitos biológicos. Uma forma de aumentar seu potencial terapêutico e reduzir algumas limitações do uso da CUR é a síntese de análogos a partir de pequenas modificações químicas na estrutura original, entretanto, mantendo sua capacidade fotossensibilizante. O objetivo desse estudo foi avaliar a ação antimicrobiana e antibiofilme de análogos de curcumina sob a influência ou não do LED sobre microrganismos de interesse endodôntico e sua influência sobre a viabilidade, proliferação e migração de fibroblastos da linhagem L-929. Uma série de compostos análogos de CUR (PCR-4 H, PCR-3 OH, PCR-4 OH, PCR-3 OCH3, PCR-4 OCH3, PCR-3 acetil, PCR-4 acetil) foram sintetizados pela metodologia de Pabon. A atividade antimicrobiana da CUR e seus análogos foi determinada pelo ensaio de Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM) sobre Streptococcus mutans, Lactobacillus casei, Actinomyces israelii, Enterococcus faecalis e Fusobacterium nucleatum, sob a ação ou não do LED InGaN (nitreto de gálio e índio, com potência de saída de 100 mW/cm², ponta do LED com área de 0,78 cm², 60 s). A curcumina e seu análogo com melhor efeito antimicrobiano (PCR-3 OH) foi avaliado sobre o biofilme inicial (72h) e maduro (1 semana) dessas espécies em microplacas e sobre biofilmes multiespécies formados em túbulos dentinários por contagem das UFC/mL e por microscopia confocal, respectivamente, sob ação ou não do LED. Também foram avaliados quanto à citotoxicidade e a capacidade de induzir proliferação e migração em fibroblastos, por meio de ensaios de metiltetrazólio, azul de tripan e azul de Coomassie, respectivamente. Os dados foram avaliados estatisticamente (p<0,05). Dos 7 análogos de curcumina sintetizados, PCR-3 OH foi o único composto que apresentou atividade bactericida quando testado sobre as bactérias de interesse endodôntico selecionadas. Seu efeito foi potencializado na presença do LED, variando entre as espécies bacterianas. A curcumina teve efeito bactericida para as espécies S. mutans, A. israelii, L. casei e F. nucleatum, e em algumas delas, foi independente do LED. Ambos os compostos reduziram o crescimento dos biofilmes iniciais ou maduros, independente do LED. Entretanto, quando irradiados, o efeito dos compostos variou de acordo com a espécie bacteriana, sendo que A. israelii e S. mutans foram os mais afetados. Ambos os compostos reduziram significativamente os biofilmes multiespécies quando comparados ao controle sem tratamento, sendo que melhor efeito foi observado para PCR-3 OH. A curcumina foi considerada citocompatível a partir de 0,039µg/mL e PCR-3 OH a partir de 0,019 µg/mL. Houve redução significativa na viabilidade celular quando os compostos foram irradiados com LED nas concentrações 0,039 e 0,019 µg/mL. O LED, dentro dos parâmetros testados, reduziu significativamente a viabilidade, a proliferação e a migração celular, independente do composto ou tempo de exposição. Conclui-se que PCR-3 OH apresentou atividade bactericida e sobre biofilmes simples e multiespécies de bactérias de interesse endodôntico superior à CUR, principalmente sob ação do LED. Entretanto, sua citocompatiblidade foi inferior à da CUR. A presença do LED afetou a viabilidade, proliferação e migração dos fibroblastos, mostrando que os parâmetros utilizados para fins antimicrobianos não foram adequados para aplicação em células eucarióticas(AU)

Endodontic treatment of young permanent teeth with pulp / periapical infections before completing rhizogenesis is still a challenge for Endodontics and Pediatric Dentistry. Scientific reports have shown that curcumin (CUR), a polyphenolic phytochemical, has several therapeutic properties, including a broad spectrum of antimicrobial action and the ability to induce cell proliferation and migration. In addition, due to its excitatory capacity in the presence of light, CUR has also been used as a photosensitizer in photodynamic therapy associated with LED (light emitting diode), promoting an increase in its biological effects. One way to increase its therapeutic potential and reduce some limitations of the use of CUR is the synthesis of analogues from small chemical modifications in the original structure, however, maintaining its photosensitizing capacity. The aim of this study was to evaluate the antimicrobial and antibiofilm action of curcumin analogues under the influence or not of LED on microorganisms of endodontic interest and their influence on the viability, proliferation and migration of L-929 fibroblasts. A series of CUR analog compounds (PCR-4 H, PCR-3 OH, PCR-4 OH, PCR-3 OCH3, PCR-4 OCH3, PCR-3 acetyl, PCR-4 acetyl) were synthesized by Pabon's methodology. The antimicrobial activity of CUR and its analogs was determined by the Minimum Concentration Inhibitory (CIM) and Minimum Bactericidal Concentration (CBM) assay on Streptococcus mutans, Lactobacillus casei, Actinomyces israelii, Enterococcus faecalis and Fusobacterium nucleatum, with or without the InGaN LED (gallium and indium nitride, with output power of 100 mW / cm², LED tip with an area of 0.78 cm², 60 sec). Curcumin and its analog with the best antimicrobial effect (PCR-3 OH) were evaluated on the initial (72h) and mature (1 week) biofilm of these species in microplates and on multispecies biofilms formed in dentinal tubules by counting CFU / mL and by confocal microscopy, respectively, under the action or not of the LED. They were also evaluated for cytotoxicity and the ability to induce proliferation and migration in fibroblasts, using methyltetrazolium, trypan blue and Coomassie blue assays, respectively. The data were evaluated statistically (p <0.05). Of the 7 curcumin analogues synthesized, PCR-3 OH was the only compound that showed bactericidal activity when tested on selected bacteria of endodontic interest. Its effect was enhanced in the presence of LED, varying between bacterial species. Curcumin had a bactericidal effect for the species S. mutans, A. israelii, L. casei and F. nucleatum, and in some of them, it was independent of the LED. Both compounds reduced the growth of the initial or mature biofilms, regardless of the LED. However, when irradiated, the effect of the compounds varied according to the bacterial species, with A. israelii and S. mutans being the most affected. Both compounds significantly reduced multispecies biofilms when compared to the untreated control, with the best effect being observed for PCR-3 OH. Curcumin was considered cytocompatible from 0.039 µg / mL and PCR-3 OH from 0.019 µg / mL. There was a significant reduction in cell viability when the compounds were irradiated with LED at concentrations of 0.039 and 0.019 µg / mL. The LED, within the parameters tested, significantly reduced cell viability, proliferation and migration, regardless of the compound or time of exposure. It is concluded that PCR-3 OH showed bactericidal activity and on simple and multispecies biofilms of bacteria of endodontic interest superior to CUR, mainly under the action of LED. However, its cytocompatibility was lower than that of the CUR. The presence of the LED affected the viability, proliferation and migration of fibroblasts, showing that the parameters used for antimicrobial purposes were not suitable for application in eukariotic cells(AU)

Content characteristics and cytotoxicity detection of silver ion products / 中华烧伤杂志

Objective@#To analyze the silver content, homogeneity, and cytotoxicity of silver-containing products.@*Methods@#(1) Five kinds of silver-containing products A, B, C, D, and E were purchased from the market, and products A, B, C, and D are liquid or gel form while product E was dressing form. The silver content of each product and the homogeneity of product E were determined by flame method. The sample number was 3. (2) Human hepatocellular carcinoma cell line (HepG2) was selected as the evaluation model. Four silver-containing products A, B, C, and D were diluted with high-glucose dulbecco′s modified eagle medium (DMEM) at multiple ratios of 1∶100, 1∶200, 1∶400, and 1∶800, and then they were used for cell culture. Cells cultured with high-glucose DMEM and high-glucose DMEM containing 20 μg/mL silver nitrate were used as blank control and positive control, respectively. The cell viability was determined by methyl thiazolyl tetrazolium assay, and each sample number was 5. (3) Four mass concentrations of 0.031 3, 0.062 5, 0.125 0, and 0.250 0 μg/mL were prepared from silver-containing product A, and then they were used to culture HepG2 cell. Cells cultured with high-glucose DMEM containing fetal calf serum and 294 μg/mL potassium dichromate were used as positive control, while those containing fetal calf serum were used as blank control. Hoechst 33258 staining method was used to detect apoptosis rate of cells. The tail moment, tail length, and the percentage of DNA in the tail of cells were observed by comet assay to evaluate DNA damage. The sample numbers were all 3. Data were processed with one-way analysis of variance and least significant difference-t test.@*Results@#The silver content of products A, B, C, and D was (256.5±1.5) μg/mL, (271.5±1.3) μg/mL, (652.4±2.6) μg/g , (330.0±2.1) μg/g, which was in accordance with labelled amount. The silver content of product E was (0.158±0.013) mg/g, and the silver content of each piece of product E was (0.125±0.017) mg/g, showing good uniformity of product E. (2) Compared with the rate of blank control, the cell survival rates of product A at the dilution ratio of 1∶100, product B at the dilution ratio of 1∶100, and product C at the dilution ratio of 1∶100 and 1∶200 were significantly reduced (t=35.506, 8.914, 37.594, 30.693, P<0.01). Compared with the rate of positive control, the cell survival rates of product A at the dilution ratio of 1∶200, 1∶400, and 1∶800, product C at the dilution ratio of 1∶400 and 1∶800, products B and D at each dilution ratio were increased significantly (t=27.537, 18.262, 18.709, 26.333, 41.762, 15.776, 19.759, 20.443, 15.715, 26.792, 24.963, 31.803, 30.537, P<0.01). (3) The apoptosis rates of cells treated by 0.250 0 μg/mL product A and positive control were (6.1±0.4)% and (62.2±3.9)% respectively, which were significantly higher than the apoptosis rate of blank control [(3.3±0.7)%, t=13.327, 30.475, P<0.05]. The apoptosis rates of cells treated by 0.031 3, 0.062 5, 0.125 0 μg/mL product A were (2.9±0.4)%, (3.1±0.4)%, and (4.2±0.9)% respectively, which were close to the apoptosis rate of blank control (t=1.181, 0.133, 1.097, P>0.05). (4) The tail moment, tail length, and tail DNA percentage of cells cultured with 0.125 0 and 0.250 0 μg/mL product A were significantly higher than those cultured with blank control (t=29.026, 51.194, 21.851, 36.138, 24.721, 50.455, P<0.05 or P<0.01). However, the tail moment, tail length, and tail DNA percentage of cells cultured with 0.031 3 and 0.062 5 μg/mL product A were close to those cultured with blank control (t=5.878, 3.429, 2.779, 1.960, 1.328, 7.763, P>0.05).@*Conclusions@#The silver content of silver-containing products meets the requirements of the labeling. The concentration of product C is higher than that of other products, leading to a greater possibility of decreasing the survival rate of HepG2 cells. It is suggested that the products A and B should be taken as reference in the concentration setting of silver ion products. The product solution with higher concentration may have higher risk of damage to cell DNA. Therefore, it is not recommended to upregulate silver content of relevant products blindly in order to achieve better antibacterial effect.

Immunotoxicity of absorbable silk fibroin biofilm on rats / 中国组织工程研究

BACKGROUND: Silk fibroin, the main component of silk fibroin biofilm, is a natural protein, but it is still a heterologous protein to the body. Its immunogenicity/toxicity is an important factor in determining the development prospects. OBJECTIVE: To evaluate the toxicity of absorbable silk fibroin biofilm and its degradation products on the rat immune system by muscle implantation experiments in rats. METHODS: Ten Wistar rats from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. were implanted with a long strip of 1 mm× 10 mm in the right gluteal muscle to observe the implant absorption and determine the implantation cycle. Seventy-two Wistar rats were assigned into control and experimental groups (n=36/group, 18 in either sexes) . A long strip of 1 mm×10 mm was implanted into the rat right gluteal muscle in the experimental group, and the control group received no implantation. Some of the rats were taken for histological examination and calculate the organ/weight ratio, at 26 weeks postoperatively. The spleen lymphocyte proliferation ability, NK cell activity, cell classification of T lymphocytes, and levels of interleukin 2 and tumor necrosis factor α were detected. Another part of the animals was taken for macrophage phagocytosis of erythrocytes cell capacity and antibody producing cell count. RESULTS AND CONCLUSION: (1) Silk fibroin biofilm was completely absorbed after implanted into the rat muscle for 26 weeks. (2) In female or male Wistar rats, the immune organs in the experimental group showed no significant changes in the appearance, weight and histological examination. There were no significant differences in the hematological indexes (hemoglobin, red blood cell count, hematocrit, blood platelet count and white blood cell count and classification) , spleen lymphocyte proliferation, NK cell activity, the ratio of T lymphocytes and their subpopulations, and the levels of interleukin 2 and tumor necrosis factor α had no significant differences between two groups (P> 0.05) . (3) In female or male Wistar rats, the macrophage phagocytosis of chicken erythrocytes cell capacity and antibody producing cell count showed no significant differences between two groups (P> 0.05) . (4) These results indicate that silk fibroin biofilm causes no immunosuppression or immunostimulation on immune organs, immune cells and immune molecules (cytokines) of rats.

Sub-chronic toxicity evaluation of Dryopteris filix-mas (L. ) schott, leaf extract in albino rats

This study evaluated the acute and sub-chronic toxicities of ethanol leaf extract of Dryopteris filix-mas. Acute toxicity and phytochemical tests on ethanol leaf extract were determined. In sub-chronic toxicity test, animals were treated with 62.5, 125, 250 and 500 mg/kg of extract every day for 90 days. Blood samples were collected via retro-orbital puncture for baseline studies and at 31, 61 and 91st days for determination of hematological, kidney and liver function parameters. Liver and kidneys were harvested for histopathology analyses on 91st day. Also, a 28 day recovery study was carried out to determine reversibility in toxicological effects. Phytochemical screening revealed the presence of tannins, phenols, flavonoids, saponins, steroids, alkaloids, terpenoids, reducing sugar and cardiac glycosides. Acute toxicity test did not show toxicity or death at 5000 mg/kg. There was significant (p<0.005) reduction in white blood cell and lymphocyte counts, significant (p<0.05) increase in some liver and kidney biomarkers as well as alterations in liver and kidney histo-architecture on 91st days in animals that were treated with 250 and 500 mg/kg extract. However, toxicities observed on 91st day were reversible in recovery studies. The leaf extract of Dryopteris filix-mas may be hepatotoxic and nephrotoxic when used for long periods

Efecto analgésico, caracterización fitoquímica y análisis toxicológico del extracto etanólico de hojas de pereskia lychnidiflora / Analgesic effect, phytochemical characterization and toxicological analysis of ethanolic extract of pereskia lychnidiflora leaves

RESUMEN Objetivo Evaluar el efecto analgésico del extracto etanólico de las hojas de Pereskia lychnidiflora, la prospección de metabolitos secundarios y el análisis toxicológico. Materiales y métodos La actividad analgésica fue evaluada mediante la prueba del ácido acético y la formalina en ratones NIH a una concentración de 30, 50 y 100 mg/kg de peso corporal, utilizando como control Ibuprofeno a 200 mg/kg y agua destilada como blanco. La prospección de metabolitos secundarios se realizó por el método de cromatografía de capa fina y la toxicidad del extracto fue evaluada in vivo según la dosis máxima de 2000 mg/kg de peso corporal. Resultados La prospección fitoquímica determinó la presencia de alcaloides, taninos, triterpenos y esteroles como mayores constituyentes químicos. Se determinó que el extracto etanólico de Pereskia lychnidiflora posee una actividad analgésica similar al Ibuprofeno. No se observaron signos de toxicidad en los ratones de experimentación y se clasifica el extracto como no tóxico con una DL50 mayor de 2000 mg/kg. Conclusión El extracto etanólico de Pereskia lychnidiflora tiene un efecto analgésico antiinflamatorio que podría estar condicionado por la presencia de alcaloides, taninos y esteroles (terpenoides) presentes en esta especie vegetal y puede ser clasificado como no tóxico.

ABSTRACT Objective To evaluate the analgesic effect of the ethanolic extract of the leaves of Pereskia lychnidiflora, the prospection of secondary metabolites and the toxicologic analysis. Materials and Methods Analgesic activity was evaluated by testing acetic acid and formalin in NIH mice at a concentration of 30, 50 and 100 mg/kg body weight, using Ibuprofen control at 200 mg/kg and distilled water as the target. Secondary metabolites were prospected using the thin layer chromatography method and the toxicity of the extract was evaluated in vivo according to the maximum dose of 2,000 mg/kg body weight. Results Phytochemical prospecting determined the presence of alkaloids, tannins, triterpenes, and sterols as major chemical constituents. The ethanolic extract of Pereskia lychnidiflora was found to have an analgesic activity similar to ibuprofen. No signs of toxicity were observed in the experimental mice and the extract is classified as non-toxic with a DL50 greater than 2,000 mg/kg. Conclusions The ethanolic extract of Pereskia lychnidiflora has an anti- inflammatory analgesic effect that could be conditioned by the presence of alkaloids, tannins, and sterols (terpenoids) present in this species and can be classified as non-toxic.

In vitro analysis of the cytotoxicity of indirect restorative materials

Abstract This study aimed to compare the cytotoxicity of the Vita AC12, Lava Ultimate, Vita Enamic and InSync indirect restorative materials. Extracts of each material were prepared by incubation for 1, 7 and 40 days, with daily washing. Human gingival fibroblasts were exposed to the extracts, and cell viability was evaluated by sequential assessment of mitochondrial activity (XTT), membrane integrity (NRU) and cell density (CVDE). Extracts of polystyrene beads and latex fragments were used as negative and positive controls, respectively. Differences between groups and experimental times were evaluated by analysis of variance. At the 24 h extraction, significant differences between the control and both Vita AC-12 and InSync were observed in the XTT assay (p<0.05), and between the control and both Enamic and Lava Ultimate, in the CVDE assay (p<0.05). AC12, Lava Ultimate, and InSync presented significantly lower cell viability than Enamic and the control group, in the NRU assay (p<0.05). The Vita Enamic and Lava Ultimate hybrid ceramic-like materials presented better biocompatibility at the 24 h extraction time point than the AC12 and InSync ceramic materials. However, a simulation of the removal of toxic components by biological fluids, conducted by using longer extraction times and daily washing, led to the absence of cytotoxicity in all the tested restorative materials. These findings can be viewed as positive for the clinical indication of these restorative materials, considering their contact with adjacent soft tissues for extended periods of time.

Resumo Este estudo teve como objetivo comparar a citotoxicidade dos materiais restauradores indiretos Vita AC12, Lava Ultimate, Vita Enamic e InSync. Extratos de cada material foram preparados por incubação por 1, 7 e 40 dias, com lavagem diária. Fibroblastos gengivais humanos foram expostos aos extratos e a viabilidade celular foi medida por avaliação sequencial da atividade mitocondrial (XTT), integridade da membrana (NRU) e densidade celular (CVDE). Extratos de esferas de poliestireno e fragmentos de látex foram utilizados como controles negativos e positivos, respectivamente. As diferenças entre os grupos e os tempos experimentais foram avaliadas por análise de variância. Na extração de 24 h, observaram-se diferenças significativas entre o controle e Vita AC-12 e InSync no teste do XTT (p<0,05) e entre o controle e os materiais Enamic e Lava Ultimate, no teste CVDE (p<0,05 ). AC12, Lava Ultimate e InSync apresentaram viabilidade celular significativamente menor do que o Enamic e o grupo controle, no ensaio NRU (p<0,05). Os materiais cerâmicos híbridos Vita Enamic e Lava Ultimate apresentaram melhor biocompatibilidade no ponto de tempo de extração de 24 h do que os materiais cerâmicos AC12 e InSync. No entanto, uma simulação de remoção de componentes tóxicos por fluidos biológicos, realizada com o uso de tempos de extração mais prolongados e lavagem diária, levou à ausência de citotoxicidade em todos os materiais restauradores testados. Esses achados podem ser vistos como positivos para a indicação clínica desses materiais restauradores, considerando seu contato com tecidos macios adjacentes por longos períodos de tempo.

An open-label, multicenter, phase I trial of a cremophor-free, polymeric micelle formulation of paclitaxel combined with carboplatin as a first-line treatment for advanced ovarian cancer: a Korean Gynecologic Oncology Group study (KGOG-3016) / 부인종양

OBJECTIVE: This phase I study aimed to determine the maximum tolerated dose (MTD) of Genexol-PM, when combined with carboplatin, as a first-line treatment in patients with advanced ovarian cancer. METHODS: This open-label, multicenter, phase I, dose-escalation study included 18 patients (median age: 59.0 years, range: 40–75 years) diagnosed with advanced epithelial ovarian cancer. All patients had measurable residual disease after debulking surgery. Patients were assigned to groups (n=6 each group) that received different doses of Genexol-PM (220, 260, and 300 mg/m², once every 3 weeks) and 5 area under the curve (AUC) carboplatin. Safety and efficacy were analyzed for each dose group. RESULTS: In this intention-to-treat population, 3 out of 18 patients dropped out of the study: 1 due to dose-limiting toxicity (DLT), 1 due to hypersensitivity, and 1 was lost during follow-up. DLTs were not reported at 220 mg/m² or 260 mg/m², but at 300 mg/m², 1 patient experienced DLT (grade 3 general pain). The MTD of Genexol-PM was not determined, but a dose of 300 mg/m² or less could be recommended for the phase II study. Most patients (73.9%) with adverse events recovered without sequelae, and no death occurred that was related to the disease or treatment. The best overall response rate was 94.1%. CONCLUSION: Genexol-PM combined with carboplatin was well tolerated as a first-line treatment, and good responses were observed in patients with advanced ovarian cancer. Based on these results, we recommended a dose of 300 mg/m² or less for a phase II study.

Teste do bafômetro: resultados e recusas de motoristas interceptados pela Operação Lei Seca no Rio de Janeiro / Breathalyzer test: results and refusals to take the test of drivers intercepted under the DUI Spot-Check Campaign in Rio de Janeiro

Resumo Estudo transversal que avaliou os resultados e as recusas ao teste do bafômetro entre motoristas interceptados pela Operação Lei Seca na capital do Rio de Janeiro e Baixada Fluminense. Realizado com base em dados cedidos pelo Governo do Estado que foram coletados durante atendimento de motoristas interceptados entre dezembro de 2013 e janeiro de 2014. Procederam-se análises descritivas da amostra e de associação entre as variáveis sexo, faixa etária e local de abordagem com as recusas e os resultados do teste do bafômetro através de um modelo de regressão logística. Dos 4756 (100%) condutores interceptados, 59 (1,2%) apresentaram resultado positivo no teste do bafômetro e 229 (4,8%) recusaram-no. Apenas a variável local de abordagem apresentou significância estatística com maiores chances de resultados positivos (OR = 4,01) e recusas ao teste (OR = 5,14) entre os condutores interceptados na Baixada Fluminense. As ações sistemáticas de fiscalização efetuadas pela Operação Lei Seca que ocorrem há mais tempo na capital parecem influenciar positivamente o comportamento de beber e dirigir dos motoristas.

Abstract Abstract This paper involved a cross-sectional study that evaluated the results and refusals to take the breathalyzer test among drivers intercepted by the Driving Under the Influence (DUI) Spot-Check Campaign (Operação Lei Seca) in the capital of Rio de Janeiro and Baixada Fluminense. It was conducted using data provided by the State Government which were collected from drivers intercepted in the months of December 2013 and January 2014. Descriptive analysis was conducted of the sample and of the association between gender, age and location variables with the result of the breathalyzer test and refusal to take the test using a logistic regression model. Of the 4756 (100%) drivers intercepted, 59 (1.2%) failed the breathalyzer test and 229 (4.8%) refused to take it. Only the location of interception variable was statistically significant with greater chances of failing (OR = 4.01) and refusal to take the test (OR = 5.14) among drivers intercepted in the Baixada Fluminense. Systematic monitoring actions taken by the DUI Spot-Check Campaign that have occurred for longer in the capital appear to have a positive impact on the drinking and driving behavior of drivers.

Progress in alternative testing strategies for human embryonic stem cell-based drug toxicity / 中国药理学与毒理学杂志

Traditional drug development and pre-clinical tests are based on animals and involve large numbers of animals,costs and long periods. Meanwhile,inter-species differences are difficult to overcome. Human embryonic stem cells (hESCs),which can self-renew and directly differentiate to types of cells,have become a new tool for toxicity alternative testing. hESC-Based alternative testing models,such as the reproductive toxicity test system,neuro development toxicity test system and metabolic model,can be used to predict target organ toxicity and toxic mechanisms of chemicals, analyze metabolic pathways and to search for potential toxicity biomarkers, when combined with omics such techiniques as metabonomics , proteomics and genomics. Therefore, hESC-based alternative testing models have extensive application to toxicology.

Development of human embryonic stem cell platforms for human health-safety evaluation / 北京大学学报(医学版)

SUMMARY Thehumanembryonicstemcells(hESCs)serveasaself-renewable,genetically-healthy, pluripotent and single source of all body cells,tissues and organs.Therefore,it is considered as the good standard for all human stem cells by US,Europe and international authorities.In this study,the standard and healthy human mesenchymal progenitors,ligament tissues,cardiomyocytes,keratinocytes,primary neurons,fibroblasts,and salivary serous cells were differentiated from hESCs.The human cellular health-safety of NaF,retinoic acid,5-fluorouracil,dexamethasone,penicillin G,adriamycin,lead ace-tate PbAc,bisphenol A-biglycidyl methacrylate (Bis-GMA)were evaluated selectively on the standar-dized platforms of hESCs,hESCs-derived cardiomyocytes,keratinocytes,primary neurons,and fibro-blasts.The evaluations were compared with those on the currently most adopted cellular platforms.Parti-cularly,the sensitivity difference of PM2.5 toxicity on standardized and healthy hESCs derived fibroblasts, currently adopted immortalized human bronchial epithelial cells Beas-2B and human umbilical vein endo-thelial cells (HUVECs)were evaluated.The results showed that the standardized hESCs cellular plat-forms provided more sensitivity and accuracy for human cellular health-safety evaluation.

Cytotoxicity analysis of electrostatically applied epoxy coating onto CO-CR alloy / Análise da citotoxidade de pintura epóxi na aplicação eletrostática sobre liga de CO-CR

Objective: To investigate the biological effect of a new method to camouflage the cobalt-chromium (CoCr) metal structure of an RPD, onto which an electrostatic paint was applied.Methods: In vitro cytotoxicity of epoxy Politherm NOBAC30C (Weg Industries SA, Santa Catarina, Brazil) in combination with polished CoCr was tested by placing it in contact with cultured human fibroblasts and comparing it with polystyrene (control surface). The cells were cultured in the presence of the test surfaces for 24, 48, 72, 94 and 120 hours. The number of viable and non-viable cells was established by manual counting. The Tukey test was used to statistically analyze cell counts between the groups.Results: The results showed that cell proliferation was similar between the groups (p =0.2174). It was observed that at 24, 48 and 72 h, there was no significant increase in cell proliferation in all groups. From 96 to 120 h, an increase in cell proliferation was observed in all groups, with no significant difference between them (p>0.05).Conclusion: The epoxy paint studied showed no cytotoxicity in vitro.

Objetivos: Analisar, biologicamente, a possibilidade do uso de pintura por aplicação eletrostática.Métodos: Por meio de testes in vitro de citotoxicidade, comparando o comportamento da tinta epóxi Politherm 30 Nobac C (Weg Indústrias S.A, Santa Catarina, Brasil) com CoCr polido e poliestireno em contato com cultura de fibroblastos humanos. Esse teste foi realizado através de contagem de células viáveis e não viáveis em tempos de 24, 48, 72, 94 e 120 horas. Para a contagem de células viáveis foi aplicada a Análise Estatística de Tukey.Resultados: Os resultados obtidos na presente pesquisa mostraram que o comportamento de crescimento celular foi estatisticamente semelhante entre grupos (p=0,2174). Observou-se que nos tempos de 24, 48 e 72 horas, não houve aumento estatisticamente significante da proliferação celular, mantendo-se o padrão para todos os grupos estudados. A partir de 96 e 120 h observamos um aumento da proliferação celular para todos os grupos estudados, sem diferenças entre os mesmos também (p>0,05). Para os resultados de células inviáveis, aplicou-se a Análise não Paramétrica de Kruskal Wallis e o teste de Dunn, devido à baixa taxa de morte celular, sem diferença estatisticamente entre os grupos (p>0,05).Conclusão: Conclui-se, portanto, que a pintura Epóxi estudada não apresentou citotoxicidade para os testes realizados in vitro.

Inspección preliminar de algunas características de toxicidad en el agua potable domiciliaria, Bogotá y Soacha, 2012 / Preliminary survey to detect toxic substances in domestic potable water, Bogotá and Soacha, 2012

Introducción. La distribución y el almacenamiento del agua después de su potabilización, pueden originar cambios significativos en su calidad, por lo que es necesario conocer las características finales del agua domiciliaria que consumen los usuarios. Objetivo. Evaluar de forma preliminar las sustancias químicas tóxicas en el agua de tanques y grifos de viviendas abastecidas por el acueducto de Bogotá. Materiales y métodos. Se hizo un estudio descriptivo en 26 viviendas localizadas en Bogotá y Soacha. Se hicieron dos muestreos: uno en época de lluvias y otro en temporada seca, y en ambos se aplicó una encuesta y se recolectaron muestras de agua de los tanques de almacenamiento y los grifos. Las muestras se sometieron a ensayos fisicoquímicos básicos, a una prueba de tamización de sustancias orgánicas e inorgánicas, y a pruebas de determinación de metales pesados y de residuos de plaguicidas organofosforados y carbamatos. Resultados. La conductividad, el color y los nitratos presentaron valores permisibles; el pH y la turbiedad tuvieron una ligera tendencia a presentar niveles altos y, el cloro residual, niveles bajos. Hubo trazas de aluminio en el 94 % de las muestras. En el período seco, 8 % de las muestras presentó compuestos orgánicos y, en el período lluvioso, 66,7 %. Solo hubo un resultado positivo para las sustancias inorgánicas. En el 11,5 % de las viviendas, el nivel de riesgo fue medio, en el 61,5 %, bajo, y en el 27 % no se registró riesgo. Conclusiones. Se evidenció deterioro de la calidad del agua domiciliaria debido a la presencia de sustancias orgánicas en tanques y sobre todo en las redes, presumiblemente a causa de la formación de biopelículas o a la presencia de polímeros orgánicos. Los niveles de aluminio cercanos al máximo aceptable pueden explicarse por los remanentes de los coagulantes utilizados durante el tratamiento.

Introduction: Significant alterations may be found in the water of Bogotá´s water supply system after its purification, specifically during its distribution and storage in home reservoirs, which makes it necessary to study the final quality of the domiciliary water consumed by users. Objective: To conduct a preliminary study of toxic chemical substances in the water supplied by Bogotá´s water supply system in samples obtained from residential reservoirs and faucets. Materials and methods: Descriptive study made in 26 homes located in Bogotá and Soacha. Two samplings were done during different seasons, each including a survey and the collection of water samples from domiciliary storage tanks and faucets. Samples were analyzed for basic physicochemical parameters, a screening test for organic and inorganic substances and the determination of heavy metals and residues of organophosphate pesticides and/or carbamates. Results: Values obtained for conductivity, color and nitrates were acceptable, pH and turbidity were slightly high while residual chlorine levels were low; aluminum traces were found in 94% of the samples; 8% of the samples analyzed during the dry season showed organic compounds, compared to 66.7% during the rainy season, and just one positive result was obtained for inorganic compounds. Consequently, a medium risk level was observed in 11.5% of homes, low risk in 61.5% and no risk in 27.0%. Conclusion: The evidence showed deterioration of the domiciliary water by organic substances present in the reservoirs as well as in the water supply piping, probably caused by the formation of biofilms or organic polymers. Aluminum levels close to the top permissible limit can be explained by the presence of residual coagulants used during water treatment.

Long-term toxicity of fully humanized anti-human tumor necrosis factor-αmonoclonal antibody for injection in cynomolgus monkeys / 中国药理学与毒理学杂志

OBJECTIVE To evaluate the long-term toxicity of fully human anti-human tumor necrosis factor-α monoclonal antibody(anti-hTNF-α FHMA)for injection in cynomolgus monkeys. METHODS Forty cynomolgus monkeys were randomly divided into 5 groups (4 males and 4 females in each group):negative control group,adalimumab 10 mg·kg-1 group,anti-hTNF-αFHMA 2,10 and 50 mg·kg-1 groups. Cynomolgus monkeys in each group were injected sc once a week for 5 consecutive times, followed by 4 weeks of recovery. During the test,general clinical observation,body mass,body temperature,electrocardiogram(ECG),hematology,coagulation function,blood biochemistry,urine, ophthalmology,immune index,and pathological changes in organs and tissues were observed. At the same time,plasma drug concentrations were detected and the toxicokinetics parameters were analyzed. RESULTS No significant toxicological changes related to drugs were observed in general clinical observation,body mass,body temperature,ECG,ophthalmic examination,blood cell counts,coagu?lation function,blood biochemistry,urine analysis,lymphocyte subsets,cytokines,serum immuno?globulin,serum complement. Neutralizing anti-drug antibody(ADA)could be detected in adalimumab group and anti-hTNF-αFHMA groups. Anti-hTNF-αFHMA showed linear dynamic characteristics in cyno?molgus monkeys. At the same dose(10 mg·kg-1),anti-hTNF-αFHMA had similar immunogenicity and kinetics characteristics to adalimumab. CONCLUSION The level of anti-hTNF-α FHMA at which no adverse effect was observed was 50 mg · kg-1,which is equivalent to 75 times clinical dosage of quasi (0.67 mg·kg-1),which suggests that anti-hTNF-αFHMA be safe in clinical use.

In vitro SCREENING ANTIBACTERIAL ACTIVITY OF Bidens pilosa LINNÉ AND Annona crassiflora MART. AGAINST OXACILLIN RESISTANT Staphylococcus aureus (ORSA) FROM THE AERIAL ENVIRONMENT AT THE DENTAL CLINIC / Triagem in vitro da atividade antibacteriana de Bidens pilosa Linné e Annona crassiflora Mart. contra Staphylococcus aureus resistente à oxacilina (ORSA) provenientes do ambiente aéreo na clínica odontológica

Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves) and Annona crassiflora (rind fruit, stem, leaves, seed and pulp) were obtained with ethanol: water (7:3, v/v) and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA) strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05). Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed) did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90) for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity.

Atualmente Staphylococcus aureus multirresistente é causa comum de infecções com altas taxas de morbidade e mortalidade mundialmente, o que direciona esforços científicos na busca de novos antimicrobianos. Neste estudo, nove extratos de Bidens pilosa (raiz, caule, flor e folhas) e de Annona crassiflora (casca do fruto, caule, folha, semente e polpa) foram obtidos com etanol:água (7:3, v/v) e suas atividades antibacteriana in vitro avaliadas através de difusão em agar e microdiluição em caldo contra 60 cepas de Oxacillin Resistant S. aureus (ORSA) e contra S. aureus ATCC 6538. Os extratos de B. pilosa e A. crassiflora inibiram o crescimento dos isolados ORSA em ambos os métodos. O extrato da folha de B. pilosa apresentou média dos diâmetros dos halos de inibição significativamente maior que a clorexidina 0,12%, contra os isolados ORSA, e os extratos foram mais ativos contra S. aureus ATCC (p < 0,05). Paralelamente, teste de toxicidade pelo método MTT e triagem fitoquímica foram avaliadas, e três extratos (raiz e folha de B. pilosa e semente de A. crassiflora) não apresentaram toxicidade. Por outro lado, as concentrações citotóxicas (CC50 e CC90) para os outros extratos variaram de 2,06 a 10,77 mg/mL. Observou-se variável presença de alcalóides, flavonóides, taninos e saponinas, apesar de total ausência de antraquinonas. Portanto, os extratos das folhas de B. pilosa revelaram boa atividade anti-ORSA e não exibiram toxicidade.

In Vitro Cytotoxicity of White MTA, MTA Fillapex® and Portland Cement on Human Periodontal Ligament Fibroblasts

The aim of this study was to compare the in vitro cytotoxicity of white mineral trioxide aggregate (MTA), MTA Fillapex® and Portland cement (PC) on human cultured periodontal ligament fibroblasts. Periodontal ligament fibroblast culture was established and the cells were used for cytotoxic tests after the fourth passage. Cell density was set at 1.25 X10 4 cells/well in 96-well plates. Endodontic material extracts were prepared by placing sealer/cement specimens (5X3mm) in 1mL of culture medium for 72 h. The extracts were then serially two-fold diluted and inserted into the cell-seeded wells for 24, 48 and 72 h. MTT assay was employed for analysis of cell viability. Cell supernatants were tested for nitric oxide using the Griess reagent system. MTA presented cytotoxic effect in undiluted extracts at 24 and 72 h. MTA Fillapex® presented the highest cytotoxic levels with important cell viability reduction for pure extracts and at ½ and » dilutions. In this study, PC did not induce alterations in fibroblast viability. Nitric oxide was detected in extract-treated cell supernatants and also in the extracts only, suggesting presence of nitrite in the soluble content of the tested materials. In the present study, MTA Fillapex displayed the highest cytotoxic effect on periodontal ligament fibroblasts followed by white MTA and PC.

Resumo O objetivo deste estudo foi comparar a citotoxicidade in vitro de agregado trióxido mineral (MTA) branco, MTA Fillapex® e cimento Portland (PC) em cultura de fibroblastos de ligamento periodontal humano. A cultura de fibroblastos de ligamento periodontal foi estabelecida e as células foram utilizadas para os testes citotóxicos após a quarta passagem. A densidade celular foi ajustada em 1,25X10 4 células/poço em placas de 96 poços. Extratos dos materiais endodônticos foram preparados por meio da inserção de corpos de prova dos cimentos (5 X 3 mm) em 1 mL de meio de cultura durante 72 h. Os extratos foram diluídos serialmente na razão de ½ e inseridos aos poços contendo as células por 24, 48 e 72 h. Ensaio de MTT foi realizado para a avaliação da viabilidade celular. O sobrenadante das células foi testado em relação à presença de óxido nítrico utilizando o sistema de reagentes de Griess. O MTA apresentou efeito citotóxico quando o extrato era aplicado sem diluição durante 24 e 72 h. O MTA Fillapex apresentou os maiores níveis de citotoxicidade com importante redução da viabilidade celular quando o extrato foi aplicado puro e em diluições de ½ e ». Neste estudo, PC não induziu alterações na viabilidade de fibroblastos. Óxido nítrico foi detectado no sobrenadante de células tratadas com os extratos e ainda nos extratos somente, o que sugere a presença de nitrito no conteúdo solúvel dos materiais testados. No presente estudo, MTA Fillapex foi o material que demonstrou o maior efeito citotóxico sobre fibroblastos de ligamento periodontal seguido do MTA branco e do PC. .

Evaluation of Acute Toxicity and Symptoms Palliation in a Hypofractionated Weekly Schedule of External Radiotherapy for Elderly Patients with Muscular Invasive Bladder Cancer

Aim To evaluate acute toxicity and symptoms palliation of a weekly hypofractionated 3DCRT schedule as radical treatment in elderly patients with organ confined bladder cancer cT1-2N0. Materials and Methods Between February 2005 and June 2011, 58 prospectively selected patients diagnosed with organ confined bladder cancer were treated with external 3DCRT (4-field arrangement). All candidates were medically inoperable, with poor performance status, and with age ranged from 75 to 88 years (median 78). A dose of 36 Gy in 6 weekly fractions was prescribed. The primary study endpoints were the evaluation of haematuria, dysuria, frequency and pain palliation as well as the acute toxicity according to the RTOG/EORTC scale: an assessment was performed at baseline, during and 3 months after radiotherapy, while the maximum reported score was taken into account. Results The gastrointestinal acute toxicities were 13/58 (22.4%) and 5/58 (5.6%), for grade I and II respectively. The genitourinary acute toxicities were 19/58 (32.7%) and 10/58 (17.2%), for grade I and II respectively. In terms of clinical outcome, 55/58 patients (94.8%) reported palliation of haematuria, while 19 out of 58 reported no change in frequency and dysuria. All patients reported significant improvement (P < 0.01) for pain, concerning the visual analogue score before and after radiotherapy. The median progression free survival was 14 months. CONCLUSIONS The incidence of patient-reported acute toxicity following weekly hypofractionated external 3DCRT is low while the symptom palliation compares very favorably with other reported outcomes. .

Role of C-Jun N-terminal kinase signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats / 中华麻醉学杂志

Objective To evaluate the role of C-Jun N-terminal kinase (JNK) signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-260 g,were randomly divided into 6 groups (n =12 each):control group (group Ⅰ),sham operation group (group Ⅱ),JNK inhibitor group (group Ⅲ),dimethyl sulfoxide (DMSO) group (group Ⅳ),lidocaine group (group Ⅴ),and JNK inhibitor and lidocaine group (group Ⅵ).Group Ⅰ received no treatment.Intrathecal catheter was placed in the subarachnoid space in group Ⅱ.SP600125 25 μg and DMSO 20 μl were injected intrathecally in Ⅲ and Ⅳ groups,respectively.In group Ⅴ,10％ lidocaine 20 μl was intrathecally injected.SP600125 25 μg was injected intrathecally and 30 min later 10％ lidocaine 20 μl was injected intrathecally in group Ⅵ.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured before intrathecal catheter was implanted (T0),before intrathecal administration (T1) and at 4,8 and 12 h and on 1,2,3,4,5 and 6 days after intrathecal administration (T2-10).At 24 h after intrathecal administration,4 rats were randomly chosen from each group and sacrificed.Their lumbar enlargements were removed for determination of phosphorylated JNK (p-JNK) expression (using Western blot) and neuronal apoptosis (by TUNEL).The apoptotic index was calculated.Results Compared with group Ⅰ,no significant difference was found in MWT and TWL in Ⅱ,Ⅲ groups and expression of p-JNK in Ⅱ and Ⅳ groups (P ＞ 0.05),MWT at T2-4,6-8 and TWL at T2-4,7 in group Ⅴ and MWT at T2-6 and TWL at T2-5 in group Ⅵ were significantly increased,the expression of p-JNK was down-regulated and the apoptotic index was decreased in group Ⅲ (P ＜ 0.05),and the expression of p-JNK was up-regulated and the apoptotic index was increased in Ⅴ and Ⅵ groups (P ＜ 0.05).Compared with group Ⅴ,MWT and TWL were significantly decreased,the expression of pJNK was down-regulated and the apoptotic index was decreased in group Ⅵ (P ＜ 0.05).Conclusion Activation of JNK signal transduction pathway is involved in spinal neurotoxicity induced by lidocaine in rats possibly through promoting neuronal apoptosis in the spinal cord.