Chemical constituents of Calotropis procera latex and ultrastructural effects on Haemonchus contortus / Constituintes químicos do látex de Calotropis procera e efeitos ultraestruturais em Haemonchus contortus

Abstract This study aimed to evaluate the anthelmintic and ultrastructural effects of Calotropis procera latex on Haemonchus contortus. C. procera latex was twice centrifuged at 10,000×g and dialyzed to obtain a fraction rich in proteins, named LP (latex protein), and at 3,000 rpm to obtain a fraction rich in secondary metabolites, named LNP (latex non-protein). Specimens of H. contortus exposed to LNP, LP and PBS in the Adult Worm Motility Test (AWMT) were submitted to scanning (SEM) and transmission (TEM) electron microscopy to verify changes in their ultrastructure. Phytochemical tests in the LNP indicated the presence of phenols, steroids, alkaloids and cardenolides. High-Performance Liquid Chromatography (HPLC) characterized the presence of the compounds gallic acid and quercetin in the LNP. The protein content in the LP was 43.1 ± 1.1 mg/mL and 7.7 ± 0.3 mg/mL in LNP. In AWMT, LNP and LP inhibited the motility of 100% of the nematodes, with LNP being more effective than LP and ivermectin more effective than both (p <0.05). Cuticle changes were observed by SEM and TEM in nematodes treated with LP and LNP. Calotropis procera latex has anthelmintic effects against H. contortus, causing damage to its cuticle and other alterations in its ultrastructure.

Resumo Este estudo objetivou avaliar os efeitos anti-helmínticos e ultraestruturais do látex de Calotropis procera sobre Haemonchus contortus. Látex de C. procera foi centrifugado duas vezes à a 10.000xg e dialisado para obter uma fração rica em proteínas, denominada proteínas do látex (LP). E centrifugado e centrifugado a 3.000 rpm, para obter uma fração rica em metabólitos secundários, denominada LNP (látex não proteico). Espécimes de H. contortus expostos à LNP, LP e PBS no Teste de Motilidade dos Nematoides Adultos (TMNA) foram submetidos a microscopia eletrônica de varredura (MEV) e de transmissão (MET), para verificar alterações em sua ultraestrutura. Testes fitoquímicos em LNP indicaram a presença de fenóis, esteroides, alcaloides e cardenolídeos. A presença dos compostos ácido gálico e quercetina em LNP foi caracterizada por Cromatografia Líquida de Alta Eficiência (CLAE). O conteúdo de proteínas em LP foi de 43,1 ± 1,1 mg/mL e de 7,7 ± 0,3 mg/mL em LNP. No TMNA, LNP e LP inibiram a motilidade de 100% dos nematoides, sendo LNP mais eficaz que LP, e a ivermectina mais eficaz que ambos (p <0,05). Alterações na cutícula de nematoides tratados com LP e LNP foram observadas por MEV e MET. O látex de C. procera apresenta efeito anti-helmíntico sobre H. contortus, causando danos à sua cutícula e outras alterações em sua ultraestrutura.

Latex proteins downregulate inflammation and restores blood-coagulation homeostasis in acute Salmonella infection

BACKGROUND Calotropis procera latex protein fraction (LP) was previously shown to protect animals from septic shock. Further investigations showed that LP modulate nitric oxide and cytokines levels. OBJECTIVES To evaluate whether the protective effects of LP, against lethal bacterial infection, is observed in its subfractions (LPPII and LPPIII). METHODS Subfractions (5 and 10 mg/kg) were tested by i.p. administration, 24 h before challenging with lethal injection (i.p.) of Salmonella Typhimurium. LPPIII (5 mg/kg) which showed higher survival rate was assayed to evaluate bacterial clearance, histopathology, leukocyte recruitment, plasma coagulation time, cytokines and NO levels. FINDINGS LPPIII protected 70% of animals of death. The animals given LPPIII exhibited reduced bacterial load in blood and peritoneal fluid after 24 h compared to the control. LPPIII promoted macrophage infiltration in spleen and liver. LPPIII restored the coagulation time of infected animals, increased IL-10 and reduced NO in blood. MAIN CONCLUSIONS LPPIII recruited macrophages to the target organs of bacterial infection. This addressed inflammatory stimulus seems to reduce bacterial colonisation in spleen and liver, down regulate bacterial spread and contribute to avoid septic shock.

Effect of the manual manipulation of composite resin with latex gloves / Efecto de la manipulación manual de resina compuesta con guantes de látex

Purpose: This in vitro study aimed to evaluate the influence of the manual manipulation of two composite resins: Filtek™ Z350XT (3M ESPE) and Herculite Précis® (Kerr), with latex gloves contaminated with powder, human saliva and alcohol, on the microhardness values. Material and Methods: Manual manipulation was evaluated using latex gloves with powder, latex gloves without powder, latex gloves without powder with saliva, latex gloves without powder with alcohol, and without hand manipulation or contaminants (control). Each resin was manually manipulated for 10 seconds and photoactivated for 20 seconds with a light intensity of 1000mW/cm2 using a VALO ­Ultradent LED light­ cured unit, and then each sample was evaluated on the microhardness Vickers tester Leitz (Wetzlar). The collected data were analyzed using Kruskal ­Wallis and Mann ­ Whitney post-test (p<0.05). Results. Microhardness values showed a significant difference between the evaluated and control groups, showing lower microhardness values in the group of latex glove with powder for Filtek™ Z350XT and the group of latex glove without powder with saliva for Herculite Précis®. Conclusion. The manual manipulation of composite resins decreases their surface microhardness.

Propósito: Este estudio in vitrotuvo como objetivo evaluar la influencia de la manipulación manual de dos resinas compuestas: Filtek ™ Z350XT (3M ESPE) y Herculite Précis® (Kerr), con guantes de látex contaminados con polvo, saliva humana y alcohol, sobre los valores de microdureza. Material y Métodos: La manipulación manual se evaluó utilizando guantes de látex con polvo, guantes de látex sin polvo, guantes de látex sin polvo con saliva, guantes de látex sin polvo con alcohol, y sin manipulación manual o contaminantes (control). Cada resina fue manipulada manualmente durante 10 segundos y fotoactivada durante 20 segundos con unaintensidad de luz de 1000mW/cm2 usando una unidad de fotocuración LED VALO - Ultradent, y luego cada muestra fue evaluada en el tester de microdureza Vickers Leitz (Wetzlar). Los datos recopilados se analizaron utilizando Kruska ­Wallis y post-test Mann­ Whitney (p<0.05). Resultados: Se observó una diferencia significativa en los valores de microdureza entre los grupos evaluados y el grupo control, con valores más bajos de microdureza en el grupo de guantes de látex con polvo para Filtek™ Z350XT y el grupo de guantes de látex sin polvo con saliva para Herculite Précis®. Conclusión. La manipulación manual de resinas compuestas disminuye su microdureza superficial.

A cysteine protease from the latex of Ficus benjamina has in vitro anthelmintic activity against Haemonchus contortus / Atividade anti-helmíntica in vitro de uma protease cisteínica do látex de Ficus benjamina contra Haemonchus contortus

Abstract Haemonchus contortus is a gastrointestinal nematode that is responsible for high mortality rates in ruminant herds. The resistance of nematodes to synthetic anthelmintics is widespread and requires a continuous search for new bioactive molecules, such as proteins. The objective of this study was to evaluate the anthelmintic potential of a protease purified from the latex of Ficus benjamina against H. contortus . Fresh latex was collected from plants via small incisions in the green stems, the rubber was removed by centrifugation, and the latex protein extract (LPE) was obtained. After LPE fractionation with ammonium sulfate and chromatography of the fraction containing the highest proteolytic activity on CM-cellulose, a cysteine protease (FbP) was purified. FbP has a molecular mass of approximately 23.97 kDa, and its proteolytic activity was stable between pH 6.0 and pH 10 and over a broad temperature range, with optimum activity at 60 °C. FbP inhibited both the development and exsheathment of H. contortus larvae, with 50% effective concentrations of 0.26 and 0.79 mg/mL, respectively. We conclude that this cysteine protease from F. benjamina latex with anthelmintic activity against H. contortus could be a promising alternative for the development of products for use in parasite control programmes.

Resumo Haemonchus contortus é um nematoide gastrintestinal, responsável por altas taxas de mortalidade em rebanhos de pequenos ruminantes. A resistência dos nematoides aos anti-helmínticos sintéticos está generalizada e requer uma busca contínua por novos compostos bioativos, como as proteínas. O objetivo deste trabalho foi avaliar o potencial anti-helmíntico da protease purificada do látex de Ficus benjamina contra H. contortus . O látex fresco foi coletado das plantas por pequenas incisões nas hastes verdes e o extrato proteico de látex (EPL) foi obtido. Após o fracionamento do EPL com sulfato de amônio e cromatografia da fração contendo a maior atividade proteolítica da CM-Celulose, uma protease cisteínica (FbP) foi purificada. A FbP tem massa molecular de cerca de 23,97 kDa, a atividade proteolítica foi estável entre pH 6,0 e pH 10 e ao longo de uma ampla faixa de temperatura, com atividade ótima a 60 °C. A FbP inibiu tanto o desenvolvimento quanto o desembainhamento das larvas de H. contortus, com 50% de inibição nas concentrações de 0,26 e 0,79 mg/mL, respectivamente. Concluímos que esta protease cisteínica do látex de F. benjamina, com ação anti-helmíntica contra H. contortus, pode ser uma alternativa promissora para o desenvolvimento de produtos a serem utilizados em programas de controle de parasitos.

Force decay evaluation of latex and non-latex orthodontic intraoral elastics: in vivo study

ABSTRACT Objective: This clinical study was conducted in order to evaluate force decay over time of latex and non-latex orthodontic intraoral elastics. Methods: Patients (n = 15) were evaluated using latex and non-latex elastics in the periods of : 0, 1, 3, 12 and 24 hours. The rubber bands were transferred to the testing machine (EMIC DL-500 MF), and force values were recorded after stretching the elastic to a length of 25mm. Paired t test was applied and analysis of variance (ANOVA) was used to evaluate the variation of force generated. LSD (Fisher's least significant difference) post-hoc test was thus employed. Results: As regards the initial forces (zero time), the values of force for non-latex elastic were slightly higher than for the latex elastic. In the subsequent times, the forces generated by the latex elastic showed higher values. Regarding the material degradation, at the end of 24 hours the highest percentage was observed for non-latex elastic. Conclusions: The latex elastics had a more stable behavior during the studied period, compared with non-latex.

RESUMO Objetivo: o objetivo deste estudo clínico foi avaliar e comparar o comportamento dos elásticos de látex e não látex quanto à perda de força ao longo do tempo. Método: os pacientes (n = 15) foram avaliados usando ambos os tipos de material (látex e não látex) em cada tempo: 0, 1, 3, 12 e 24 horas. Os elásticos foram transferidos para a máquina de ensaios mecânicos (EMIC DL-500 MF) e os valores de força foram registrados após a distensão dos elásticos a uma distância de 25 mm. Foi aplicado o teste t pareado, e a análise de variância (ANOVA) foi realizada para verificar a variação das forças geradas em todos os tempos estudados. Em seguida, o teste post-hoc LSD (Fisher's least significant difference) foi aplicado. Resultados: quanto às forças iniciais (tempo zero), os valores de força dos elásticos não látex foram ligeiramente maiores do que dos elásticos de látex. Nos tempos subsequentes, as forças geradas pelos elásticos de látex apresentaram valores superiores às geradas pelos elásticos não látex. Em relação à degradação do material, ao fim do período de 24 horas, o maior percentual foi observado pelos elásticos não látex. Conclusões: os elásticos de látex apresentaram comportamento mais estável durante o período de estudo, em comparação aos não látex.

Involvement of mast cells, CD68+ and VEGF+ expressions in response to Himatanthus drasticus commercial latex in mice wound healing model / Envolvimento de mastócitos, expressão de CD68+ e VEGF+ em resposta ao látex comercial de Himatanthus drasticus em modelo de cicatrização em camundongos

This study aimed to evaluate Himatanthus drasticus latex in a mice wound healing experimental model. Animals were divided into four groups (n=7) according to the treatments: GI - saline 0.9% (control), GII - mineral oil (vehicle), GIII - H. drasticus commercial latex (HdCL) and GIV - H. drasticus mixed isolated fraction (MIF, 1 mg/mL). The treatments were applied topically once daily, 50 µL for 14 consecutive days. Macroscopic lesions were evaluated, considering parameters such as swelling, redness, granulation tissue and reepithelialization. VEGF+, CD68+ expressions and mast cells (Toluidin blue stain) were evaluated. HdCL induced higher contraction and exuberant granulation tissue (P > 0.05). HdCL showed a mild inflammatory process while MIF induced intense infiltrate inflammatory predominantly by lymphocytes, vascular congestion, bleeding and did not presented full reepithelialization. Reorganization of collagen fibers (red picrosirius stain) was observed. CD68+ expression and mast cells were presented as moderate, intense and mild in GI, GIII and GIV, respectively. Neovascularization occurred in all groups, while VEGF+ expression was intense in MIF in relation to HdCL. We concluded that HdCL presents wound healing potential, through modulation of mast cells, CD68+ and VEGF+ expressions that can be associated to triterpenes presence according MIF isolated from HdCL.(AU)

Objetivou-se avaliar o látex de Himatanthus drasticus em feridas induzidas experimentalmente em camundongos. Os animais foram divididos em quatro grupos (n=7): GI - salina 0,9% (controle), GII - óleo mineral (veículo), GIII - látex comercial de H. drasticus (HdCL) e GIV - fração isolada mista de H. drasticus (MIF, 1mg/mL). Os tratamentos foram aplicados topicamente uma vez ao dia (50µL), durante 14 dias consecutivos. Lesões macroscópicas, as expressões de VEGF+, CD68+ e a participação dos mastócitos (coloração azul de toluidina) foram avaliadas. HdCL induziu maior contração e tecido de granulação exuberante (P >0,05). HdCL induziu leve processo inflamatório enquanto MIF promoveu intenso infiltrado inflamatório predominantemente linfocítico, congestão vascular, hemorragia e reepitelização parcial. Observou-se reorganização das fibras colágenas (coloração picrosírius). A expressão de CD68+ e os mastócitos apresentaram-se moderados, intensos e leves em GI, GIII e GIV, respectivamente. A neovascularização foi observada em todos os grupos, enquanto a expressão de VEGF+ foi mais intensa em MIF em relação a HdCL. Conclui-se que HdCL apresenta potencial de cicatrização por meio da modulação dos mastócitos e das expressões de CD68+ e VEGF+, o que pode estar associado à presença de triterpenos de acordo com MIF isolada de HdCL.(AU)

Proteomic analysis of latex from Jatropha curcas L. stems and comparison of two classic proteomic sample isolation methods: the phenol extraction and the TCA/acetone extraction

Background: Jatropha curcas is a wide-spreading latex-rich biodiesel plant with high oil content in seeds that have always been under intense studies. However, studies are lacking on the latex component that is considered rich in proteins with potentially important physiological functions and secondary metabolites that are a promising source for new drugs. The proteomic analysis, which would be the first step to study these substances, was hampered by the presence of interfering components. Phenol extraction and Trichloroacetic acid (TCA)/ acetone extraction, two major plant proteomic isolation methods, were used and compared in this study. Results: We identified 459 proteins from the J. curcas latex proteome using the combination of the two extraction techniques. Although more number of latex proteins were identified by the phenol extraction (401 proteins vs. 123 proteins by the TCA/acetone extraction), only 65 proteins were commonly isolated by both methods. Analysis of the biochemical properties revealed that relatively more number of lower isoelectric point (pI) proteins were isolated by the TCA/acetone method (pI mode: 4.79, 6.51 for phenol). Moreover, GO, COG, and KEGG analyses showed that certain classes/categories/pathways annotated more number of proteins than others, and most of them had proportionally comparable protein counts by both the methods, however, with exemplified exceptions. Conclusions: A large number of proteins were found and exclusively identified by either method, indicating that a better proteome coverage of plant samples in a similar context needs the combined use of multiple isolation methods. In addition, the core biological function of the latex may be uncovered by certain GO, COG, and KEGG classes/categories/pathways that annotate more proteins.

Evaluation of wound healing treated with latex derived from rubber trees and Aloe Vera extract in rats

ABSTRACT PURPOSE: To compare the use of latex derivative and Aloe vera extract to wound healing. METHODS: Twenty one rats were randomly divided into three groups and each one had a wound made by incision. The treatment consisted in: derivative of latex (GL), Aloe vera extract (GA) and saline solution (GC). The wound area was measured on the 7th, 14th and 21st days and macroscopic and microscopic evaluation were done. RESULTS: The comparison between the measurements of the wounds presented statistical difference in GC and GA from the 7th day of evaluation and GL from the 14th day. The extent of the wound was significantly smaller by the 7th day in GL. Histologically, in GL, the neovascularization was significant on the 7th, 14th and 21st days. On the 21st day the scar was large and little mature. In GA and GC, the findings were similar on the 7th, 14th and 21st days with a slight better organization of skin and collagen on the 21st in GA. CONCLUSIONS: Statistical analysis did not allow for the definition of the best topical agent. The latex had the highest angiogenesis, but a possible foreign body granuloma. Aloe vera has revealed a healing process adequated temporally in histology.

Aislamiento y caracterizacion parcial de fracciones con actividad ribonucleasa a partir del latex de calotropis procera (Aiton) W.T. Aiton y pedilanthus tithymaloides (L.) poit / Isolation and partial characterization of fractions with ribonuclease activity from latex of calotropis procera (Aiton) W.T. Aiton and pedilanthus tithymaloides (L.) poit

Con el objetivo de aislar y caracterizar parcialmente las enzimas ribonucleasas (RNasas) contenidas en el látex de Calotropis procera y Pedilanthus tithymaloides, se colectaron muestras de plantas adultas. Las proteínas solubles fueron extraídas con acetato de sodio y centrifugación a 16.000 x g durante 15 min y fraccionadas por cromatografía de intercambio iónico. Se estimó la masa molecular a través de ecuaciones de regresión lineal. Se realizaron pruebas de glicosilación. En ambas especies, las proteínas con actividad RNasa presentaron una masa molecular entre 28 y 30 kDa. No existe evidencia de proteínas glicosiladas en el látex de C. procera. En P. tithymaloides la RNasa es una proteína glicosilada.

In order to isolate and characterize partially ribonucleases (RNases) enzymes contained in the latex from Calotropis procera and Pedilanthus tithymaloides, samples were collected from mature plants. Soluble proteins were extracted with sodium acetate and centrifugation at 16,000 xg for 15 min and fractionated by ion exchange chromatography. Molecular mass was estimated by linear regression equations. Glycosylation tests were conducted. In both species, proteins with RNase activity showed a molecular mass between 28 and 30 kDa. No evidence of glycosylated proteins in latex from C. procera. In P. tithymaloides, RNase may be a glycosylated protein.

Elaboración de un compuesto farmacológico a base de látex de Carica papaya con actividad cicatrizante evaluada en modelo experimental in vivo en ratas Wistar, con amputación de colas en la Facultad de Ciencias Médicas de julio - octubre 2012 / Preparation of a pharmacological compound based on latex of Carica papaya with healing activity evaluated in experimental model in vivo in Wistar rats, with amputation of tails at the Faculty of Medical Sciences of July - October 2012

En una herida por trauma siempre hay un proceso inflamatorio, durante este proceso, simultáneamente se desencadena un proceso proteolítico para eliminar el tejido muerto y depositar el nuevo tejido, se propone acelerar este último utilizando una enzima proteolítica. Se analizó la existencia de las enzimas proteolíticas comercialmente disponibles y se comparó su actividad con la de látex de papaya que contiene altas concentraciones de una enzima proteolítica llamada papaína. Se concluyó que látex de papaya tiene varias ventajas importantes sobre las enzimas disponibles comercialmente. Se desarrolló método analítico de monitoreo de la actividad enzimática de látex de papaya, tanto como el método de su purificación. Se comparó la concentración de látex en distintos tejido de Carica papaya, encontrando entre los resultados que las frutas son las fuentes más ricas en látex, y que el compuesto presenta estabilidad térmica y en medios con pH acido. Con todo esto se logró concluir que la aplicación de un preparado de látex de Carica papaya en heridas por amputación quirúrgica de la cola de las ratas acelera el proceso de cicatrización, el grupo intervención en relación a los grupos control.

Influence of different beverages on the force degradation of intermaxillary elastics: an in vitro study

Objective: The aim of this study was to evaluate in vitro the effects of frequently ingested beverages on force degradation of intermaxillary elastics. Material and Methods: One hundred and eighty 1/4-inch intermaxillary elastics (TP Orthodontics) were immersed into six different beverages: (1) Coca-Cola®; (2) Beer; (3) Orange juice; (4) Red wine; (5) Coffee and (6) artificial saliva (control). The period of immersion was 15 min for the first and second cycles and 30 min for the third to fifth cycles. Tensile forces were read in a tensile testing machine before and after the five immersion cycles. One-way repeated measures ANOVA was used to identify significant differences. Results: Force degradation was seen in all evaluated groups and at all observation periods (p<0.05). A greater degree of degradation was present at the initial periods, decreasing gradually over time. However, no statistically significant differences were seen among groups at the same periods, showing that different groups behaved similarly. Conclusion: The chemical nature of the evaluated beverages was not able to influence the degree of force degradation at all observation periods.

Effect of five brands of latex gloves on the setting time of polyvinyl siloxane putty impression materials.

Addition silicone impression materials have been used as impression material for more than 20 years. Although they are among the most expensive impression materials, they became popular during the past decade as they have excellent physical properties. Prevention of infection is an important aspect in dental treatment since dental professionals are routinely exposed to the wide variety of microorganisms present in saliva. Gloves are the most common protective measure used during dental treatment. The gloves are mostly made of latex. In this study, we examine how the setting time of three types polyvinyl putty materials were affected by the use of five different brands of latex gloves and one brand of vinyl gloves. Each material was first mixed without wearing gloves according to the manufacturer's instructions. After the stipulated mixing time, the setting time was measured using the Vicat needle. The setting time is measured from the time of mixing till the time that the needle does not produce any indentation on the surface of the material. The putty material was then mixed with gloved hands (using the five different brands of latex gloves in turn) and the setting time was measured. Then the material was mixed with washed gloved hands, and the setting time was measured again. Finally, the material was mixed with vinyl gloved hands and the setting time was measured. The following conclusions were drawn from the study: Reprosil™ and Express™ showed significant variation in the setting time with the latex gloved hands. There was no significant variation in the setting time when material was mixed with unwashed vs washed gloved hands. Vinyl gloves did not significantly affect the setting time of any of the putty impression materials.

Protective effect of aqueous suspension of dried latex of Calotropis procera against oxidative stress and renal damage in diabetic rats

Calotropis species have been used in the traditional medicinal system for the treatment of diseases of the liver and abdomen. In view of the antioxidant and anti-hyperglycemic properties of an aqueous suspension obtained from the dried latex of Calotropis procera, the present study was carried out to evaluate its efficacy in affording protection against alloxan induced changes in rat kidney. A single intraperitoneal injection of alloxan (150 mg/kg) in rats produced hyperglycemia within 3 days and altered kidney functions over a period of 90 days. Daily oral administration of the aqueous suspension (100 and 400 mg/kg) in diabetic rats produced anti-hyperglycemic effect that was comparable to that of glibenclamide (10 mg/kg). Unlike glibenclamide, the aqueous suspension did not increase the serum insulin levels in diabetic rats. However, it produced a marked reduction in the levels of urinary glucose and protein and normalized the renal tissue levels of thiobarbituric acid-reactive substances (TBARS) and glutathione (GSH) in diabetic rats and the effect was comparable to that of glibenclamide. The protection afforded by the aqueous suspension was also evident from the histological analysis of the renal tissue. Our study shows that by exhibiting antioxidant and anti-hyperglycemic property the aqueous suspension of dried latex of C. procera affords protection against the complications associated with diabetes.

Effects of sulfur-based hemostatic agents and gingival retraction cords handled with latex gloves on the polymerization of polyvinyl siloxane impression materials

OBJECTIVES: This study investigated the possible interactions between three addition silicone materials (Express®, Aquasil Ultra® and Adsil®), three hemostatic agents (ferric sulfate, StatGel FS®; aluminum sulfate, GelCord®; and aluminum chloride, Hemostop®) and gingival retraction cords previously handled with latex gloves to determine whether direct contact with medicaments or indirect contamination by latex in conditions similar to those found in clinical practice inhibit or affect the setting of the impression materials. MATERIAL AND METHODS: A portable device for the simultaneous test of several specimens was specifically developed for this study. Polymerization inhibition was analyzed by examination of the impressions and the molded surface. Ten trials were performed for each addition silicone material used in the study, at a total of 240 study samples. RESULTS: All the samples tested (N=240) were nonreactive regardless of the type of combination used. CONCLUSIONS: Aluminum sulfate, ferric sulfate and aluminum chloride hemostatic solutions did not show any inhibitory potential on the addition silicone samples under study, and there were no changes in polymerization as a result of contact between addition silicone and retraction cords handled with latex gloves.

In vitro cytotoxicity of latex orthodonticelastics / Citotoxicidad de los elásticos ortodònticos de látex

Natural latex does not fall into the category of materials known to be entirely inoffensive. The objective of the present in vitro study is to test the hypothesis that there is no difference in the cytotoxicity between natural latex elastics of different colours. The present article compared different latex intra-oral elastics (5/16 = 7.9 mm). The sample was divided into four groups according to their manufacturer: Group N (Natural latex elastic, Morelli), Group R (Red colour elastic, Morelli) Group Y (Yellow colour elastic, Morelli) and Group G (Green colour elastic, Morelli). Cytotoxicity assays were performed by using cell culture medium containing L-929 line cells (mouse fibroblast). The cytotoxicity was evaluated by using the “dyeuptake” test, which was employed at two different moments (1 and 24 h). Data were compared by analysis of variance (ANOVA) and Tukey’s test (p<0.05). The results showed a significant difference (p<0.05) between the groups N, R, Y, G and the negative cytotoxicity control at 1 and 24 h (p<0.05), it did not have presented significant difference between the groups N, R, Y, G tested (p>0.05) at 1 and 24 h. Morelli intra-oral elastics were found to be highly cytotoxic, regardless of their colour and immersion time.

El látex natural no entra en la categoría de materiales que se sabe del todo inofensivo. El objetivo del presente estudio in vitro es poner a prueba la hipótesis de que no hay diferencia en la citotoxicidad entre elásticos de látex natural de diferentes colores. El presente artículo compara diferentes elásticos intraorales de látex (5/16 =7,9 mm). La muestra se dividió en cuatro grupos según su fabricante: Grupo N (elástico látex, Morelli), Grupo I (elástico de color rojo, Morelli) Grupo Y (elástico de color amarillo, Morelli) y el Grupo G (elástico color verde, Morelli). Pruebas de citotoxicidad se realizaron mediante el uso de medio de cultivo celular que contiene líneas celulares L-929 (fibroblastos de ratón). La citotoxicidad se evaluó mediante el test “dye-uptake”, que se empleó en dos momentos diferentes (1 y 24 h). Los datos se compararon mediante análisis de varianza (ANOVA) y test de Tukey (p<0,05). Los resultados mostraron una diferencia significativa (p<0,05) entre los grupos N, R, Y, G y la negativa citotoxicidad del control en 1 y 24 h (p<0,05), no han presentado diferencias significativas entre los grupos N, R , Y, G probado (p>0,05) en 1 y 24 h. Elásticos intraorales Morelli resultaron ser altamente citotóxicos, independientemente de su color y tiempo de inmersión.

In vitro antimicrobial activity of four ficus carica latex fractions against resistant human pathogens [antimicrobial activity of ficus carica latex]

Methanolic, hexanoi'c, chloroformic and ethyl acetate extracts of Ficus carica latex were investigated for their in vitro antimicrobial proprieties against five bacteria species and seven strains of fungi. The green fruit latex was collected from Chott Mariam Souse, Middle East coast of Tunisia. The antimicrobial activity of the extracts was evaluated and based respectively on the inhibition zone using the disc - diffusion assay, minimal inhibition concentration [MIC] for bacterial testing and the method by calculating inhibition percentage [1%] for fungi-inhibiting activities. The methanolic extract had no effect against bacteria except for Proteus mirabilis while the ethyl acetate extract had inhibition effect on the multiplication of five bacteria species [Enterococcus fecalis, Citobacter freundei, Pseudomonas aeruginosa, Echerchia coll and Proteus mirabilis]. For the opportunist pathogenic yeasts, ethyl acetate and chlorophormic fractions showed a very strong inhibition [100%]; methanolic fraction had a total inhibition against Candida albicans [100%] at a concentration of 500 micro g/ml and a negative effect against Cryptococcus neoformans. Microsporum canis was strongly inhibited with methanolic extract [75%] and totally with ethyl acetate extract at a concentration of 750 micro g/ml. Hexanoi'c extract showed medium results

Observações sobre abelhas sem ferrão (Hymenoptera: meliponini) coletoras de látex em Abricó do Pará Mammea americana (L.) Jacq. (Clusiaceae), Manaus, Amazonas, Brasil / Observations on stingless bees (Hymenoptera: meliponini) collecting of latex in mammy Mammea americana (L.) Jacq. (Clusiaceae), Manaus, state of Amazonas, Brazil

O conhecimento das espécies de plantas utilizadas pelas abelhas sem ferrão é primordial para a manutenção das mesmas. O objetivo deste trabalho foi verificar a coleta de látex em Mammea americana (Clusiaceae) e os padrões de abundância dos visitantes. O estudo foi conduzido no Bosque da Ciência, Instituto Nacional de Pesquisas da Amazônia, em Manaus-AM. A comunidade de abelhas foi representada por Trigona williana e T. recursa. A espécie presente em maior abundância foi T. recursa (89,4%). Estas abelhas coletam o látex para proteger o ninho contra possíveis inquilinos e predadores, devido a sua ação antimicrobiana.

The knowledge regarding the plant species used by the stingless bees for their survival is a fundamental factor for their maintenance. The objective of this study is to observe collecting latex in Mammea americana tree (Clusiaceae) and abundance patterns of visitors. The study was carried out in the “Bosque da Ciência,” Instituto Nacional de Pesquisas da Amazônia, Manaus-AM. The bee communities were represented by Trigona williana and Trigona recursa. The latter presented the highest abundance (89.4%). These bees probably collect this substance in order to protect their nest against other possible tenants and predators, since it acts as an antimicrobial agent.

Potential of laticifer fluids for inhibiting Aedes aegypti larval development: evidence for the involvement of proteolytic activity

It has been shown previously that the laticifer fluid of Calotropis procera (Ait.) R.Br. is highly toxic to the egg hatching and larval development of Aedes aegypti L. In the present study, the larvicidal potential of other laticifer fluids obtained from Cryptostegia grandiflora R.Br., Plumeria rubra L. and Euphorbia tirucalli L. was evaluated. We attempted to correlate larvicidal activity with the presence of endogenous proteolytic activity in the protein fraction of the fluids. After collection, the fluids were processed by centrifugation and dialysis to obtain the soluble laticifer protein (LP) fractions and eliminate water insoluble and low molecular mass molecules. LP did not visibly affect egg hatching at the doses assayed. LP from Cr. grandiflora exhibited the highest larval toxicity, while P. rubra was almost inactive. E. tirucalli was slightly active, but its activity could not be correlated to proteins since no protein was detected in the fluid. The larvicidal effects of LP from C. procera and Cr. grandiflora showed a significant relationship with the proteolytic activity of cysteine proteinases, which are present in both materials. A purified cysteine proteinase (papain) from the latex of Carica papaya (obtained from Sigma) was similarly effective, whereas trypsin and chymotrypsin (both serine proteinases) were ineffective. The results provide evidence for the involvement of cysteine proteinase activity in the larvicidal action of some laticifer fluids. C. procera is an invasive species found in areas infested with Ae. aegypti and thus could prove useful for combating mosquito proliferation. This is the first report to present evidence for the use of proteolytic enzymes as chemical agents to destroy Ae. aegypti larvae.

Molecular and biochemical evaluation of genetic effect of Calotropis procera (Ait.) latex on Aspergillus terreus (Thom).

The present study was conducted to observe the genotoxic effect of latex of Calotropis procera. Dense conidial suspensions of Aspergillus terreus Thorn were treated with different concentrations of Calotropis latex and assayed for percentage survival, auxotrophic mutants, DNA and total protein contents and further subjected to RAPD and SDS-PAGE tests. It was found that latex of Calotropis procera had potent lethal and mutagenic activities. Percentage survival decreased as concentration or time of exposure increased. Frequency of auxotrophic mutants increased with increase in concentration or exposure time. Most auxotrophic mutants were amino acid requiring mutants. DNA and total protein contents of each mutant was significantly lower than wild type of Aspergillus terreus. RAPD demonstrated polymorphic genetic bands which were the electrophoretic products of PCR for all mutants compared with the wild type strain. SDS-PAGE results expressed a polymorphism of protein bands as well. All these results indicated the mutagenicity of latex of Calotropis procera.

Clinical relevance of trans 1.4-polyisoprene aging degradation on the longevity of root canal treatment

This in vivo study investigated the time of degradation of root filling material (trans 1,4-polyisoprene) retrieved from endodontically treated teeth and correlated the occurrence of degradation with the longevity of endodontics. Thirty-six root-filled teeth with different filling times (2 to 30 years) and with and without periapical lesions were selected. All teeth presented clinical indication for root canal retreatment. The association among filling time, presence of periapical lesion and root filling material degradation was investigated. Root filling samples were retrieved from the root canals using a Hedströ m file without solvent. The trans 1,4-polyisoprene was isolated by root filling solubilization in chloroform followed by filtration and centrifugation. GPC and FT-IR were the analytical techniques utilized. Degradation of trans 1,4-polyisoprene occurred with time, as a slow process. It is an oxidative process, and production of carboxyl and hydroxyl groups in the residual polymer were observed. Statistically significant decrease of molar mass was observed after 5 (p=0.0001) and 15 (p=0.01) years in teeth with and without periapical lesion, respectively. Bacteria participated in polymer degradation. Gutta-percha aging was proven an important factor for the long-term success of endodontic treatment. The findings of the present study showed that, after 15 years, polymer weight loss may decrease the capacity of the filling mass to seal the root canal space and prevent re-infection, thus compromising significantly the longevity of root canal therapy.

Este estudo in vivo avaliou a degradação do material obturador e a influência deste fator na longevidade do tratamento endodôntico. Foram selecionados 36 pacientes (3-30 anos) com canais tratados endodonticamente, com e sem lesões periapicais, e indicação de retratamento endodôntico. Foi investigada a associação entre o tempo de tratamento, presença de lesão periapical e a degradação do material obturador. O material obturador foi removido com uma lima Hedströ em sem uso de solvente. O polímero trans 1,4- poliisopreno foi isolado do material obturador através de solubilização em clorofórmio, seguido de filtragem e centrifugação. GPC e FT-IR foram os métodos analíticos utilizados. A degradação do trans 1,4- poliisopreno foi observada com o tempo, sendo um processo lento e oxidativo, com formação de grupos carboxílicos e hidroxilas no polímero residual. Após 5 (p=0,0001) e 15 (p=0,01) anos, em dentes com e sem lesões periapicais, respectivamente, houve decréscimos significantes na massa molar do material obturador. A infecção bacteriana participa no processo de degradação do polímero. O envelhecimento da guta-percha é um fator que influencia o sucesso a longo prazo do tratamento endodôntico. Após 15 anos, a longevidade do tratamento pode ser significantemente afetada pela redução da capacidade de selamento causada pela perda de massa molar do polímero, permitindo a reinfecção do sistema de canais radiculares.