Insecticidal potential of organic extracts of Calotropis procera to Spodoptera frugiperda

This study evaluated the toxic effects of organic extracts of Calotropis proceraleaves on the survival, development, and reproduction of Spodoptera frugiperda. Solutions of crude methanol extract and hexane and methanol fractions of C. proceraleaves were added at 1.15% and 2.14% concentrations to the artificial diet of S. frugiperda. The mortality and duration of larval and pupal phases, weights of female and male pupae, deformations of pupae and adults, the reduction of adults able to reproduce, pre-oviposition and oviposition periods, the number of postures per female, and the fecundity and fertility of S. frugiperdafemales were also evaluated. The extracts harmed the survival, development, and reproduction of S. frugiperda. The ingestion of extracts and fractions by caterpillars affected adults by decreasing the oviposition period, the number of postures, fecundity, and fertility. The crude MeOH extract at a2.14% concentration harmed the evaluated parameters of the insect, except for pupal mortality, female pupae weight, and pre-oviposition period. The MeOH fraction at 2.14% caused a 50.0% mortality of caterpillars and 16.0% deformation in pupae and 33.0% inadults, reducing by 72.0% the population able to reproduce. The MeOH fraction at the 2.14% concentration caused 25.0% and 38.0% of pupal mortality and deformation, respectively. Calotropis procerahas promising insecticidal properties for a biological insecticide, a convenient and sustainable strategy for protecting plants against S. frugiperda.

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.

Agronomic performance of carrot fertilized with roostertree [Calotropis procera (Ait.) R. BR.] in two growing seasons / Desempenho agronômico da cenoura adubada com flor-de-seda [Calotropis procera (Ait.) R. BR.] em duas épocas de cultivo

Because vegetables have a fast production cycle, they require fertilization in quantities and at ideal times to obtain maximum productivity, and green manure is an alternative practice to the use of mineral fertilizers. This study was conducted to evaluate the effects of different amounts of roostertree [Calotropis procera (Ait.) R. Br.] biomass and its incorporation times in two growing seasons, in the agronomic performance of carrots. The experiment was conducted on an experimental farm at the Universidade Federal Rural de Pernambuco (UFRPE) in the autumn-winter period (March-July 2012) and the spring-summer period (September-December 2012). The experimental design was a randomized block design with three replications. The treatments were arranged in a factorial 4 x 4 design; the first factor consists of four amounts of roostertree biomass (5.4, 8.8, 12.2 and 15.6 t ha-1 on a dry basis), and the second factor consists of four times of incorporation of this manure into the soil (0, 10, 20 and 30 days before sowing the carrots). The carrot cultivar used was Brasília. The following traits were evaluated: plant height, number of leaves per plant, root dry mass, total and commercial yield of roots. The best agronomic performance of carrot cultivar Brasília was found with the amount of roostertree biomass of 15.6 t ha-1, in the time of incorporation into the soil of 10 days before seeding. The cultivation in the autumn-winter showed higher total and commercial productivities of carrot roots fertilized with roostertree.

As hortaliças, por possuírem ciclo rápido de produção, demandam adubações em quantidades e momentos ideais para a obtenção da máxima produtividade, sendo a adubação verde uma prática alternativa ao uso de fertilizantes minerais. O presente estudo foi realizado com o objetivo de avaliar os efeitos de diferentes quantidades de biomassa de flor-de-seda [Calotropis procera (Ait.) R. Br.] e seus tempos de incorporação, em duas épocas de cultivo, no desempenho agronômico da cenoura. O experimento foi conduzido em campo experimental pertencente à Universidade Federal Rural de Pernambuco (UFRPE), nos períodos outono-inverno (março a julho de 2012) e primavera-verão (setembro a dezembro de 2012). O delineamento experimental utilizado foi em blocos casualizados, com três repetições. Os tratamentos foram arranjados em esquema fatorial 4 x 4, com o primeiro fator constituído por quatro quantidades de biomassa de flor-de-seda (5,4; 8,8; 12,2 e 15,6 t ha-1 em base seca), e o segundo fator, por quatro tempos de incorporação ao solo deste adubo (0, 10, 20 e 30 dias antes da semeadura da cenoura - DAS). A cultivar de cenoura utilizada foi a Brasília. Foram avaliados a altura de plantas, número de folhas por planta, massa seca de raízes, produtividades total e comercial de raízes. O melhor desempenho agronômico da cenoura cv. Brasília foi obtido com a quantidade de flor-deseda de 15,6 t ha-1, no tempo de incorporação ao solo de 10 dias antes da semeadura. O cultivo no outono-inverno promoveu maiores produtividades total e comercial às raízes de cenoura adubada com flor-de-seda.

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.

Bioatividade dos extratos aquosos de plantas às larvas da mosca-das-frutas, Ceratitis capitata (Wied. ) / Bioactivity of aqueous extracts of plants to the fruit fly larvae, Ceratitis capitata (Wied. )

O objetivo deste trabalho foi avaliar a toxicidade dos extratos aquosos de plantas às larvas recém-eclodidas de Ceratitis capitata. Folhas de algodão-de-seda (Calotropis procera), citronela (Cymbopogon nardus), juazeiro (Ziziphus joazeiro), pinha (Annona squamosa),pinhão-manso (Jatropha curcas), noni (Morinda citrifolia) e tingui (Magoniapubescens) foram coletadas, secas e moídas em moinho de facas até a obtenção de um pó. Tais pós foram adiciona dos em água destilada para a preparação dos extratos aquosos (10%, p/v). As suspensões contendo os compostos hidrossolúveis foram filtradas, a fim de se conseguir os extratos que foram adicionados à dieta artificial das larvas da mosca. À dieta controle, adicionou-se somente água destilada. Os ensaios foram realizados em DIC, com quatro tratamentos e um controle. Todos os tratamentos foram constituídos por 10 repetições, cada uma com 10 larvas recém-eclodidas de C. capitata. Foram avaliadas a mortalidade larval e a eficiência de controle (E%) dos extratos aquosos às larvas. Os extratos de juazeiro, tingui e pinhão-manso são os mais tóxicos e eficientes no controle das larvas de C. capitata, causando elevadas mortalidades.(AU)

The objective of this study was to evaluate the toxicity of aqueous plant extracts to neonate larvae of Ceratitis capitata. Silk cotton (Calotropis procera), citronella grass (Cymbopogon nardus), joazeiro (Ziziphus joazeiro),sugar apple (Annona squamosa), jatropha (Jatropha curcas), noni (Morinda citrifolia) and tingui (Magonia pubescens) were collected, dried, and milled in a knife mill in order to obtain a powder. These powders were added in distilled water to prepare aqueous extracts (10%, w/v). The suspension containing the hydrosoluble compounds was filtered to get the extracts, which were added to the artificial diet of fruit fly larvae. Only distilled water was added in the control diet. Assays were carried out in DIC, with four treatments (aqueous extracts) and one control. All treatments consisted of 10 repetitions, each containing 10 newly hatched larvae ofC. capitata. Larval mortality and control efficiency (E%) of aqueous extracts to the larvae were evaluated. The joazeiro, tingui and jatropha extracts are the most toxic and effective in controlling larvae of C. capitata, causing high mortality.(AU)

Development of standard operating procedure [SOP] of a unani pharmacopoeial formulation

In various Unani pharmacopoeias thousands of formulations are present which have been formulated by eminent physicians of Unani System of Medicine. The majority of these formulations are based on plant origin drugs and their efficacy may get altered as a result of seasonal variation, use of different drying methods, their storage conditions and microbiai contamination. To avoid these alterations, standardization of drugs is necessary especially in case of compound formulations the use of Standard Operating Procedures [SOPs] for their manufacturing is essential. It will help to prepare a formulation repeatedly with the same quality. The present study was designed to develop the SOP of a Unani pharmacopoeial formulation i.e., Habbc Gule Aak [HG]. Total eight batches of HG were prepared with different conditions i.e., binder, particle size, time of drying and temperature of drying'. All the eight batches of HG were evaluated thrice for the hardness, friability and disintegration time. The batch with minimum friability, hardness nearest to standard value and minimum disintegration time as per pharmacopoeias was selected as final batch and all conditions regarding powder size, binder, temperature of drying and time of drying used in the preparation of frnal batch were fixed as its Standard Operating Procedure. Out of eight batches the Batch No. 2 with 150 micron particle size [100 mesh sieve], mucilage of 30% w/w samaghe arabi [binder], dried at 80°C for 150 minutes was selected as final batch. The final batch showed minimum friability [0.05 +/- 0.01], hardness nearest to the standard value [4.59 +/- 0.30] and considerable disintegration time [33.67 +/- 0.33]. The resulting SOP will be helpful in setting the pharmacopoeial standards of HG and the methodology of the work will help in the development of SOPs for other Unani formulations too

Citotoxicidad in vitro de extractos laticiferos de Calotropis procera (Aiton) W.T. Aiton y Pedilanthus tithymaloides (L.) Poit / In vitro cytotoxicity of laticifers extracts from Calotropis procera (Aiton) WT and Pedilanthus tithymaloides (L) Poit

In order to assess the anticancer action of extracts obtained by latex from Calotropis procera and Pedilanthus tithymaloides, samples were collected from adult plants. Soluble proteins were extracted with 16 uL of 50 mM sodium acetate pH 5/ug integral latex and centrifugation at 16,000 x g for 15 min, the supernatant was named "latex crude extract" (LCE). The "latex methanolic extract" (LME) was obtained on dried latex. Both extracts were tested in vitro by cytotoxic and cytostatic activity in Jurkat cell cultures. Cellular viability, proliferation, necrosis and apoptosis were evaluated. LCE and LME of C. procera were found with cytotoxic and cytostatic activity after 24 incubation hours (p < 0,05) with doses from 1ug/mL. The LCE and LME of P. tithymaloides presented cytotoxic effect (p < 0,05) from 50 ug/mL and from 1ug/mL, respectively.

Con el objetivo de evaluar el potencial anticanceroso de extractos de látex de Calotropis procera y Pedilanthus tithymaloides se colectaron muestras de plantas adultas. Las proteínas solubles fueron extraídas con 16 uL de acetato de sodio 50 mM pH 5/ug de látex integral y centrifugación a 16.000 x g durante 15 min, denominándose al sobrenadante “extracto crudo de látex” (ECL). El “extracto metanólico de látex” (EML) se obtuvo sobre látex deshidratado. Ambos extractos fueron probados en su actividad citotóxica y citostática in vitro sobre cultivos de células Jurkat. Se realizaron estudios de viabilidad, proliferación, necrosis y apoptosis celular. El ECL y el EML de C. procera presentaron actividad citotóxica y citostática después de 24 y 48 horas de incubación (p < 0,05) con dosis desde 1 ug/mL. Los ECL y EML de P. tithymaloides presentaron efectos citotóxicos (p < 0,05) a partir de 50 ug/mL y desde 1 ug/mL respectivamente.

Biometria de frutos e sementes e germinação de Calotropis procera Aiton (Apocynaceae) / Biometry of fruits and seeds and germination of Calotropis procera Aiton (Apocynaceae)

Essa pesquisa teve como objetivos avaliar a biometria dos frutos e sementes de Calotropis procera, bem como estabelecer as condições de luz, temperatura e substrato adequados à germinação de sementes dessa espécie. A biometria dos frutos foi determinada numa amostra de 100 unidades, coletados ao acaso, no município de Mossoró-RN, constituindo-se nas determinações de comprimento, largura, espessura e peso de frutos e sementes. Dessas sementes, numa amostra de 100 unidades retiradas aleatoriamente, determinaram-se comprimento, número de sementes por fruto, teor de água e peso de mil sementes. Foram avaliadas as seguintes variáveis: germinação e tempo médio de germinação. Para análise empregou-se o delineamento experimental inteiramente casualizado em arranjo fatorial 6x3x2 sendo seis temperaturas (20, 25, 30, 35, 20-30 e 25-30 ºC), três substratos (areia, papel toalha e mata-borrão) e duas condições de luminosidades (presença e ausência de luz), com seis repetições de 50 sementes. Os dados biométricos foram analisados através da estatística descritiva e para os caracteres germinação e tempo médio de germinação utilizou-se análise de variância e teste de Tukey através do SISVAR 4.5. Concluiu-se que há variabilidade para os caracteres relacionados às dimensões e pesos de frutos de flor-de-seda; o número de sementes por fruto é elevado, favorecendo a disseminação da espécie. Recomenda-se que as contagens inicial e final do teste de germinação sejam realizadas aos 5 e 10 dias após a semeadura, respectivamente. A temperatura constante de 30 ºC, alternada de 25-30 ºC e os substratos areia e papel toalha são condições adequadas para a germinação e as sementes são indiferentes à luminosidade.

This research aimed to evaluate the biometry of fruits and seeds Calotropis procera and to establish the optimum light, temperature and substrate conditions for seeds germination. It was evaluated length, width, thickness and weight of fruits and seeds, number of seeds per fruit, moisture content and weight of 1000 seeds. Evaluated the following variables: germination and the germination average time. The experimental design was completely randomized with a factorial arrangement 6x3x2, using six temperatures (20, 25, 30, 35, 20-30 e 25-30 ºC), three substrates (sand, towel paper and blotting paper) and two light conditions (presence and absence), with six replications of 50 seeds. It was concluded that there is variability for characters related to fruit size and weight of silk-flower, the number of seeds per fruit is high, favoring the spread of the species, widely distributed in the semiarid region. It is recommended that the initial and final counts of the germination test are performed at 5 and 10 days after sowing, respectively. The constant temperature of 30 °C, alternating 25-30 °C and the substrates sand and paper roll were the most appropriate conditions for germination and the seeds are indifferent to light.

Acaricidal activity of synthesized titanium dioxide nanoparticles using Calotropis gigantea against Rhipicephalus microplus and Haemaphysalis bispinosa

OBJECTIVE@#To assess the acaricidal activity of titanium dioxide nanoparticles (TiO2 NPs) synthesized from flower aqueous extract of Calotropis gigantea(C. gigantea) against the larvae of Rhipicephalus (Boophilus) microplus [R. (B.) microplus] and the adult of Haemaphysalis bispinosa (H. bispinosa).@*METHODS@#The lyophilized C. gigantea flower aqueous extract of 50 mg was added with 100 mL of TiO(OH)2 (10 mM) and magnetically stirred for 6 h. Synthesized TiO2 NPs were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), and Energy dispersive X-ray spectroscopy (EDX). The synthesised TiO2 NPs were tested against the larvae of R. (B.) microplus and adult of H. bispinosa were exposed to filter paper impregnated method.@*RESULTS@#XRD confirmed the crystalline nature of the nanoparticles with the mean size of 10.52 nm. The functional groups for synthesized TiO2 NPs were 1 405.19, and 1 053.45 cm(-1) for -NH2 bending, primary amines and amides and 1 053.84 and 1 078.45 cm(-1) for C-O. SEM micrographs of the synthesized TiO2 NPs showed the aggregated and spherical in shape. The maximum efficacy was observed in the aqueous flower extract of C. gigantea and synthesized TiO2 NPs against R. (B.) microplus (LC50=24.63 and 5.43 mg/L and r(2)=0.960 and 0.988) and against H. bispinosa (LC50= 35.22 and 9.15 mg/L and r(2) = 0.969 and 0.969), respectively.@*CONCLUSIONS@#The synthesized TiO2 NPs were highly stable and had significant acaricidal activity against the larvae of R. (B.) microplus and adult of H. bispinosa. This study provides the first report of synthesized TiO2 NPs and possessed excellent anti-parasitic activity.

Evaluation of the toxicological effects manifested after long term administration of aqueous Calotropis Procera plant extract in male and female rabbits

Treatment with aqueous extract of Calotropis Procera was examined only for short periods. So, The present study was designed to examine the influences of long term administrations of aqueous extract of Calotropis Procera on male and female rabbits. The extract was daily oral administered to thirty adult rabbits divided into two sets each set contain 3 groups [5/each] for a period of 8 weeks. The doses administered were 80 and 40 mg/kg body weight as high and low doses for both male and female test groups. Percentage of body weight change, serum biochemistry [Alanine amino transferases [ALAT], aspartate amino transferases [ASAT], Alkaline phosphatase, total protein, albumin, globulin, A/G ratio, urea and creatininre], hematological parameters [PCV, WBCs, RBCs, Platelets and Hb] and the blood indices [M.C.V., M.C.H. and M.C.H.C.] were examined. Results showed that there was no mortality or clinical signs observed among the treatment and the control groups. No significant changes in the percentage of body weight gain, hematological parameters, blood indices has been noticed in all treated groups Highly significant decreases in serum total protein and albumin in male and female rabbits has been recorded at high doses. While the globulin and A/G ratio revealed no significant changes. Serum ASAT, urea and creatinine levels were highly significant increases [P < 0.01] and the activity of ALAT and ALP was significant increases [P < 0.05] in male and female rabbits at high doses only. Some toxicological effects were observed after administration of aqueous leaves extract of Calotropis Procera at high doses in the long term for male and females rabbits

Efeito do extrato aquoso do algodão de seda (Calotropis procera Aiton) sobre a eficiência reprodutiva do carrapato bovino / Effect of aqueous extract of silk cotton (Calotropis procera Ait. R.Br.) on the reproductive performance of Rhipicephalus microplus

O carrapato Rhipicephalus microplus é um dos principais parasitos dos rebanhos bovinos. Diferentes grupos de carrapaticidas sintéticos têm sido empregados para o controle desse ácaro, contudo, o uso constante desses produtos tem selecionado populações resistentes. Na tentativa de minimizar esses impactos tem-se buscado avaliar fitoterápicos para o controle desse parasito. Assim, objetivou-se analisar a eficácia in vitro do extrato aquoso do algodão de seda em diferentes concentrações no controle da postura e eclodibilidade dos ovos do R. microplus. Foram avaliados os efeitos das concentrações 1, 5, 10, 25, 50, 75 e 100% do extrato aquoso obtido juntamente com controles com água destilada ou com cipermetrina, 12,5 mg L-1, associada a organofosforado, 150 mg L-1, sobre a eficiência reprodutiva do carrapato. Foi realizado o biocarrapaticidograma sendo que a eficácia acaricida das concentrações 5, 25 e 100% foram superiores àquela observada para o produto comercial utilizado, que correspondeu a 93,98%, indicando a importância de futuras pesquisas para avaliar o potencial toxigênico e a eficácia in vivo dessa planta para o controle do R. microplus .

The tick Rhipicephalus microplus is one of the main parasites of bovine herds. Different groups of synthetic acaricides have been used to control this mite; however, the constant use of these products has selected resistant populations. In an attempt to minimize these impacts, researchers have tried to evaluate herbal remedies to control this parasite. Therefore, the aim of this study was to analyze the in vitro efficacy of aqueous extract of silk cotton at different concentrations to control the hatchability of R. microplus eggs. The effects of the concentrations 1, 5, 10, 25, 50, 75 and 100% aqueous extract together with controls with distilled water or with cypermethrin, 12.5 mg L-1, associated with organophosphate, 150 mg L-1, were assessed on the reproductive efficiency of the tick. Immersion test was performed and the acaricide efficacy of the 5, 25 and 100% concentrations were higher than that obtained for the commercial product, which corresponded to 93.98%, indicating the importance of further research to assess the toxigenic potential and the in vivo efficacy of this plant to control R. microplus .

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.

Phenytoin-induced Toxicity in the Postnatal Developing Cerebellum of Wistar Rats, Effect of Calotropis procera on Histomorphometric Parameters / Toxicidad Inducida por Fenitoína en el Desarrollo Postnatal del Cerebelo de Ratas Wistar, Efecto de Calotropis procera sobre los Parámetros Histomorfométricos

The role of methanolic leaf extracts of Calotropis procera in phenytoin-induced toxicity on histomorphometric variables in the postnatal developing cerebellum of Wistar rat was studied. Pregnant rats were treated orally with 50 mg/kg phenytoin in pre and post natal life and 300 mg/kg methanolic leaf extract of Calotropis procera 1 hour prior to phenytoin administration. 200 mg/kg vitamin C (standard antioxidant) was also administered orally 1 hour prior to phenytoin treatment. The control animals received water. Standard diet of rat pellets and water were provided ad libitum. At the end of the experiment, the offspring of days 1, 7, 14, 21, 28 and 50 post partum, five per group were sacrificed by cervical dislocation. The cerebellum of all groups were dissected out and processed for histomorphometric studies. The results showed in the developing cerebellum of phenytoin treated animals, a delayed cell maturation in the external granular layer, reduction of the molecular layer, astrocytic gliosis and loss of Purkinje cells on day 50 postpartum. Administration of extracts of Calotropis procera and vitamin C though reversed these changes when compared with the phenytoin treated group, but not significantly when compared with the control. In conclusion, supplementation with methanolic extracts of Calotropis procera reduced the rate at which phenytoin induced toxicity in the postnatal developing cerebellum of Wistar rat.

Fue estudiado el rol de los extractos metanólicos de las hojas de Calotropis procera en la toxicidad inducida por fenitoína sobre las variables histomorfométricas en el desarrollo postnatal del cerebelo de ratas Wistar. Ratas preñadas fueron tratadas por vía oral con 50 mg/kg de fenitoína durante la vida pre y post natal. Además, fue administrado, por vía oral, una hora antes del tratamiento con fenitoína 300 mg/kg de extracto metanólico de las hojas de Calotropis procera y 200 mg/kg de vitamina C (antioxidante estándar). Los animales control recibieron agua. Una dieta estándar de pellets para rata y agua se proporcionaron ad libitum. Al final del experimento, 5 crías por grupo de 1, 7, 14, 21, 28 y 50 días post parto, fueron sacrificadas por dislocación cervical. El cerebelo de todos los animales de los diferentes grupos fueron disecados y procesados para el estudio histomorfométrico. Los resultados mostraron en el desarrollo del cerebelo de los animales tratados con fenitoína un retraso en la maduración de células en la capa granular externa, reducción de la capa molecular, gliosis astrocitaria y pérdida de las células de Purkinje en el día 50 post parto. La administración de extractos de Calotropis procera y vitamina C, aunque invirtieron estos cambios, en comparación con los grupos tratados con fenitoína, no fueron significativos en comparación con el control. En conclusión, la suplementación con extractos metanólicos de Calotropis procera redujo la velocidad a la que la fenitoína induce toxicidad en el desarrollo postnatal del cerebelo de ratas Wistar.

Evaluation of antitumour activity of Calotropis gigantea L. root bark against Ehrlich ascites carcinoma in Swiss albino mice

OBJECTIVE@#To investigate experimentally the possible antitumor effect of methanol extract (ME) of Calotropis gigantea L. (C. gigantean) root bark and its petroleum ether (PEF) and chloroform (CF) soluble fractions against Ehrlich ascites carcinoma (EAC) in Swiss albino mice.@*METHODS@#The effects of ME (10 and 20 mg/kg), PEF (40 and 80 mg/kg) and CF (20 and 40 mg/kg) on the growth of EAC and life span of EAC bearing mice were studied. Hematological profile and biochemical parameters (SALP, SGPT and SGOT) were also estimated.@*RESULTS@#Results of in vivo study showed a significant decrease in viable tumor cell count and a significant increase of life span in the ME and CF treated group compared to untreated one. The life span of ME and CF treated animals was significantly (P<0.05) increased by 43.90% (20 mg ME/kg) and 57.07% (40 mg CF/kg). ME and CF brought back the hematological parameter more or less normal level. ME and CF also restored the altered levels of serum alkaline phosphatase (SALP) and serum glutamate oxaloacetate transaminase (SGOT).@*CONCLUSIONS@#Methanol extract (ME) of C. gigantea root bark and its chloroform soluble fraction (CF) possesses significant antitumor activity.

Molecular characterization and transcriptome profiling of expansin genes isolated from Calotropis procera fibers

The Calotropis procera seed fibers provide an excellent model system to study the genes involved in fiber elongation, fineness and strength. Expansins constitute one of the important gene families involved in plant cell expansion and other cell wall modification processes. Four homologs of Expansin A gene i.e. CpEXPA1, CpEXPA2, CpEXPA3 and CpEXPA4 were isolated from the cDNA library obtained from fast growing Calotropis procera fibers. These homologs represented typical Expansin A family. Each of them had two conserved domains including GH45 like domain and the putative polysaccharide binding domain. The deduced amino acid sequences of the homologs indicated three conserved motifs: i) eight cysteine residues at N-terminus, ii) four tryptophan residues at C-terminus and iii) a Histidine-Phenylalanine-Aspartate motif in the center of the sequence. The presence of N-terminal signal peptide consisting of hydrophobic amino acids and a transmembrane region in all these expansin isoforms suggests their cotranslational insertion into the endoplasmic reticulum and then transportation to the cell wall by secretory pathway. The relative quantification of the four expansins in root, stem, fiber and leave tissues indicated that the transcripts of CpEXPA1, CpEXPA2, CpEXPA3 and CpEXPA4 are variably transcribed in these tissues. The lowest transcription of all the four Expansin A isoforms was observed in elongating roots indicating that root tissue might be having specific expansins other than those confined to air grown organs.

In vitro and in vivo antiproliferative activity of Calotropis procera stem extracts

The cytotoxic potential of stem organic extracts from Calotropis procera (Asclepiadaceae) was firstly evaluated against cancer cell lines by MTT assay. Subsequently, samples considered cytotoxic were tested for antimitotic activity on sea urchin egg development and for in vivo antiproliferative activity in mice bearing Sarcoma 180 tumor. Among the five extracts (hexane, dichloromethane, ethyl acetate, acetone and methanol), ethyl acetate and acetone extracts displayed higher cytotoxic potential against tumor cells, with IC50 ranging from 0.8 to 4.4 μg/mL, while methanolic extract was weakly cytotoxic. Cytotoxic extracts also exhibited cell division inhibition capacity by antimitotic assay, revealing IC50 values lower than 5 μg/mL. In the in vivo antitumor assessments, ethyl acetate- and acetone-treated animals showed tumor growth inhibition ratios of 64.3 and 53.1 percent, respectively, with reversible toxic effects on liver and kidneys. Further studies are in progress in order to identify C. procera cytotoxic compound(s) and to understand the mechanism of action responsible for this tumor-decreasing potential.

O potencial citotóxico de extratos orgânicos do caule de Calotropis procera (Asclepiadaceae) foi primeiramente avaliado frente a linhagens de células tumorais através do ensaio de MTT. Aquelas amostras consideradas citotóxicas foram sub-sequentemente testadas para atividade antimitótica sobre o desenvolvimento de ovos de ouriço-do-mar e para atividade antiproliferativa in vivo em camundongos transplantados com tumor Sarcoma 180. Dentre os cinco extratos estudados (hexano, diclorometano, acetato de etila, acetona e metanol), os extratos acetato de etila e acetona mostraram maior potencial citotóxico contra células tumorais, com CI50 variando de 0,8 to 4,4 μg/mL, enquanto o extrato metanólico revelou ser fracamente citotóxico. s extratos citotóxicos também exibiram capacidade de inibição da divisão celular com valores de CI50 menores que 5 μg/mL. Nas avaliações antitumorais in vivo, os animais tratados com os extratos acetato de etila e acetona mostraram taxas de inibição do crescimento tumoral de 64,3 e 53,1 por cento, respectivamente, com efeitos tóxicos reversíveis sobre o fígado e os rins.

Ocular toxicity by latex of Calotropis procera (Sodom apple).

We report the spectrum of ocular toxicity following accidental inoculation of latex of Calotropis procera (Sodom apple) in 29 eyes between January 2003 and December 2006. All patients presented with sudden painless dimness of vision with photophobia. Twenty-five (86%) patients had initial visual acuity of less than 20/60. All eyes had conjunctival congestion and mild to severe corneal edema with Descemet's folds. Three (10%) eyes had an epithelial defect, nine (31%) had iridocyclitis, and seven (24%) had associated secondary glaucoma. After treatment with topical corticosteroids, antiglaucoma agents, cycloplegics, hypertonic saline and tears supplements, 27 (93%) eyes recovered completely within 3-14 days. After three months, 17 (74%) out of 23 eyes showed a significant low endothelial cell count compared to the normal fellow eye ( P 0.001). The latex of Calotropis procera causes significant ocular morbidity which may be preventable by simple health education. The long-term effect on corneal endothelium has to be studied further.

Calotropis procera -induced keratitis.

Calotropis procera produces copious amounts of latex, which has been shown to possess several pharmacological properities. Its local application produces intense inflammatory response. In the 10 cases of Calotropis procera -induced keratitis reported here, the clinical picture showed corneal edema with striate keratopathy without any evidence of intraocular inflammation. The inflammation was reversed by the local application of steroid drops.

Antimicrobial and Cytotoxic Activity of Di-(2-ethylhexyl) Phthalate and Anhydrosophoradiol-3-acetate Isolated from Calotropis gigantea (Linn.) Flower

A phytochemical study on the flowe r of Calotropis gigantea (Linn.) using silica gel column chromatography and preparative thin layer chromatography, led to the first time isolation of Di-(2-ethylhexyl) phthalate (compound 1) and anhydrosophoradiol-3-acetate (compound 2). The structures of these compounds were confirmed by spectroscopic analyses (IR, HRTOFMS and NMR). The antibacterial and antifungal activities of ethyl acetate extract, compound 1 and compound 2 were measured using the disc diffusion method. Ethyl acetate extract and compound 1 presented better results than compound 2. The minimum inhibitory concentrations (MICs) of the extract and compounds were found to be in the range of 16~128 microg/ml. The cytotoxicity (LC50) against brine shrimp nauplii (Artemia salina) were also evaluated and found to be 14.61 microg/ml for ethyl acetate, 9.19 microg/ml for compound 1 and 15.55 microg/ml for compound 2.