In vitro anti-plasmodial activity of three selected medicinal plants that are used in local traditional medicine in Amhara region of Ethiopia.

BACKGROUND: The plants Aloe weloensis, Lepidium sativum, and Lobelia gibberoa have been used in Ethiopian folklore medicine to treat various diseases including malaria. METHOD: The in vitro anti-plasmodial activity of the three crude extracts was evaluated using parasite lactate dehydrogenase assay against the chloroquine (CQ)-sensitive D10 and the chloroquine (CQ)-resistant W2 strains. RESULT: The methanolic extract of L. gibberoa roots showed the highest in vitro anti-plasmodial effect against both D10 and W2 Plasmodium falciparum strains with IC50 value of 103.83 ± 26.17 µg/mL and 47.11 ± 12.46 µg/mL, respectively. However, the methanolic extract of L. sativum seeds and the leaf latex of A. weloensis were not active with an IC50 value > 200 µg/mL against both D10 and W2 strains. CONCLUSION: The methanolic extract of L. gibberoa roots showed a promising in vitro anti-plasmodial activity against the CQ-sensitive (D10) and CQ-resistant (W2) strains of P. falciparum. Thus, the anti-plasmodial activity of this plant partly justifies and may also support the traditional use against malaria. However, the methanolic extract of L. sativum seeds and the leaf latex of A. weloensis did not exert suppressive activity on the growth of P. falciparum strains.

Euphorbia bicolor (Euphorbiaceae) Latex Extract Reduces Inflammatory Cytokines and Oxidative Stress in a Rat Model of Orofacial Pain.

Recent studies have reported that the transient receptor potential V1 ion channel (TRPV1), a pain generator on sensory neurons, is activated and potentiated by NADPH oxidase-generated reactive oxygen species (ROS). ROS are increased by advanced oxidation protein products (AOPPs), which activate NADPH oxidase by upregulating Nox4 expression. Our previous studies reported that Euphorbia bicolor (Euphorbiaceae) latex extract induced peripheral analgesia, partly via TRPV1, in hindpaw-inflamed male and female rats. The present study reports that E. bicolor latex extract also can evoke analgesia via reduction of oxidative stress biomarkers and proinflammatory cytokines/chemokines in a rat model of orofacial pain. Male and female rats were injected with complete Freund's adjuvant (CFA) into the left vibrissal pad to induce orofacial inflammation, and mechanical allodynia was measured by the von Frey method. Twenty-four hours later, rats received one injection of E. bicolor latex extract or vehicle into the inflamed vibrissal pad. Mechanical sensitivity was reassessed at 1, 6, 24, and/or 72 hours. Trigeminal ganglia and trunk blood were collected at each time point. In the trigeminal ganglia, ROS were quantified using 2',7'-dichlorodihydrofluorescein diacetate dye, Nox4 protein was quantified by Western blots, and cytokines/chemokines were quantified using a cytokine array. AOPPs were quantified in trunk blood using a spectrophotometric assay. E. bicolor latex extract significantly reduced orofacial mechanical allodynia in male and female rats at 24 and 72 hours, respectively. ROS, Nox4, and proinflammatory cytokines/chemokines were significantly reduced in the trigeminal ganglia, and plasma AOPP was significantly reduced in the trunk blood of extract-treated compared to vehicle-treated rats. In vitro assays indicate that E. bicolor latex extract possessed antioxidant activities by scavenging free radicals. Together our data indicate that the phytochemicals in E. bicolor latex may serve as novel therapeutics for treating oxidative stress-induced pain conditions.

Protection afforded by methanol extract of Calotropis procera latex in experimental model of colitis is mediated through inhibition of oxidative stress and pro-inflammatory signaling.

Calotropis procera, a latex producing plant is known to possess medicinal properties including its beneficial effect in gastrointestinal disorders. The anti-inflammatory effect of its latex in various experimental models is noteworthy and in light of this the present study was carried out with an objective to evaluate its efficacy in ulcerative colitis, an inflammatory condition of the colon. Colitis was induced in rats by acetic acid and the rats were divided into four groups where one group served as experimental control and the other groups were treated with two doses of methanol extract of dried latex of C. procera (MeDL; 50 and 150 mg/kg) and mesalazine (MSZ; 300 mg/kg). The study also included normal control (NC) group for comparison of various parameters related to colon like macroscopic changes, ulcer score, adherent mucus content, weight/length ratio, small intestinal transit, oxidative stress and inflammatory markers, tissue histology and immunoreactivity of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and nuclear factor kappa beta (NFκB) subunit p65. Treatment of colitic rats with MeDL produced a significant reduction in colonic mucosal damage as revealed by macroscopic and microscopic evaluation and normalization of tissue levels of oxidative stress markers and pro-inflammatory mediators. The protection afforded by MeDL was also evident from its restorative effect on tissue histology and expression of COX-2, iNOS and NFκB(p65). This study shows that by targeting oxidative stress and NFκB(p65) mediated pro-inflammatory signaling, the latex of C. procera affords protection in colitis and its effect was comparable to that of mesalazine. This study suggests that latex of C. procera could serve as a promising therapeutic option for treating inflammatory conditions of the colon.

Exploring hemostatic and thrombolytic potential of heynein - A cysteine protease from Ervatamia heyneana latex.

ETHNOPHARMACOLOGICAL RELEVANCE: The latex of Ervatamia heyneana (Wall.) T. Cooke plant has been used for wound healing and various skin diseases by Indian tribes and folklore. AIM OF THE STUDY: To validate the scientific basis of heynein - a key protease of Ervatamia heyneana, in hemostasis and wound healing process. MATERIALS AND METHODS: The latex from E. heyneana was processed and subjected to two step purification. The purified heynein was assayed for proteolytic activity using casein as substrate and also attested by zymography. The inhibition studies confirmed the nature of heynein. Pure fibrinogen was used for fibrinogenolytic activity and citrated plasma was used for coagulant and fibrinolytic activities. The edema inducing action and hemorrhagic activity of heynein were assessed on mice model. RESULTS: The purified heynein exhibited proteolytic activity, which was confirmed by caseinolytic assay and zymography. The inhibition studies confirmed heynein to be a cysteine protease. Heynein showed complete hydrolysis of all the three subunits of human fibrinogen (Aα, Bß, γ). It exhibited strong pro-coagulant activity by reducing plasma clotting time from 248 to 39s at 40µg concentration. Heynein cleaved α polymer subunit in fibrin clot and did not induce edema and hemorrhage in mice models. The non-hemorrhagic nature was supported with histopathological studies of skin samples. CONCLUSION: Heynein displays strong pro-coagulant action associated with fibrin(ogen)olytic activity. This provides basis for the observed pharmacological action of Ervatamia heyneana and thereby justifies its use in folk medicine.

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

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.

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.

Effect of Hevea brasiliensis latex sap gel on healing of acute skin wounds induced on the back of rats.

OBJECTIVE: to evaluate the effect of topical delivery of latex cream-gel in acute cutaneous wounds induced on the back of rats. METHODS: we subjected sixteen rats to dermo-epidermal excision of a round dorsal skin flap, with 2.5cm diameter. We divided the animals into two groups: Latex Group: application of cream-gel-based latex throughout the wound bed on postoperative days zero, three, six and nine; CONTROL GROUP: no treatment on the wound. Photographs of the lesions were taken on the procedure day and on the 6th and 14th postoperative days, for analyzing the area and the larger diameter of the wound. We carried out euthanasia of all animals on the 14th postoperative day, when we resected he dorsal skin and the underlying muscle layer supporting the wound for histopathological study. RESULTS: there was no statistically significant difference in the percentage of wound closure, in the histopathological findings or in the reduction of the area and of the largest diameter of the wounds among the groups studied on the 14th postoperative day. CONCLUSION: according to the experimental conditions in which the study was conducted, latex cream-gel did not interfere in the healing of acute cutaneous wounds in rats.

Effect of Hevea brasiliensis latex sap gel on healing of acute skin wounds induced on the back of rats / Efeito do gel da seiva do látex da Hevea brasiliensis na cicatrização de lesões cutâneas agudas induzidas no dorso de ratos

Objective : to evaluate the effect of topical delivery of latex cream-gel in acute cutaneous wounds induced on the back of rats. Methods : we subjected sixteen rats to dermo-epidermal excision of a round dorsal skin flap, with 2.5cm diameter. We divided the animals into two groups: Latex Group: application of cream-gel-based latex throughout the wound bed on postoperative days zero, three, six and nine; Control group: no treatment on the wound. Photographs of the lesions were taken on the procedure day and on the 6th and 14th postoperative days, for analyzing the area and the larger diameter of the wound. We carried out euthanasia of all animals on the 14th postoperative day, when we resected he dorsal skin and the underlying muscle layer supporting the wound for histopathological study. Results : there was no statistically significant difference in the percentage of wound closure, in the histopathological findings or in the reduction of the area and of the largest diameter of the wounds among the groups studied on the 14th postoperative day. Conclusion : according to the experimental conditions in which the study was conducted, latex cream-gel did not interfere in the healing of acute cutaneous wounds in rats.

Objetivo : avaliar o efeito da administração tópica do gel-creme de látex em feridas cutâneas agudas induzidas no dorso de ratos. Métodos : dezesseis ratos foram submetidos à excisão dermoepidérmica de retalho cutâneo dorsal, circular com 2,5cm de diâmetro. Os animais foram distribuídos em dois grupos, um experimental e outro controle: Grupo Látex- aplicação em todo o leito da ferida do látex em base gel-creme no período zero, no terceiro, no sexto e no nono dias pós-operatórios; Grupo Controle- sem nenhum tratamento sobre a ferida. Foram feitas fotografias das lesões no dia da operação, no sexto e no 14º dia pós-operatório, para análise de área e do maior diâmetro da ferida. Realizou-se a eutanásia de todos os animais no 14º dia pós-operatório. Ressecou-se a pele dorsal e o plano muscular subjacente contendo a ferida para estudo histopatológico. Resultados : não houve diferença estatisticamente significante no percentual de fechamento, nos achados histopatológicos ou na redução da área e do maior diâmetro das feridas, entre os grupos estudados no 14º dia pós-operatório. Conclusão : nas condições experimentais em que o estudo foi realizado, o gel-creme de látex não interferiu na cicatrização de feridas cutâneas agudas em ratos.

EFFECTS OF TOPICAL TREATMENT WITH EUPHORBIA TIRUCALLI LATEX ON THE SURVIVAL AND INTESTINAL ADHESIONS IN RATS WITH EXPERIMENTAL PERITONITIS / EFEITOS DO TRATAMENTO TÓPICO COM O LÁTEX DA EUPHORBIA TIRUCALLI NA SOBREVIDA E NAS ADERÊNCIAS INTESTINAIS DE RATOS COM PERITONITE EXPERIMENTAL

Background: The use of plants of the family Euphorbiaceae, particularly Euphorbia tirucalli (avelós) has been popularly widespread for treating a variety of diseases of infectious, tumoral, and inflammatory. Aim: To demonstrated antimicrobial and immunomodulatory effects of these extracts, evaluating the effect of a topical treatment with an aqueous solution of avelós latex on the survival and on intestinal adhesions in rats with experimental peritonitis. Methods: Peritonitis was induced in 24 Wistar rats, that were randomized into four groups of six as follows: (1) Control group (n=6), no treatment; (2) Antibiotic group (n=6), treatment with a single intramuscular dose of antibiotic Unasyn; (3) Saline group (n=6), the abdominal cavity was washed with 0.9% saline; and (4) E.tirucalli group (n=6), the abdominal cavity was washed with E. tirucalli at a concentration of 12 mg/ml. The animals that died were necropsied, and the time of death was recorded. The survivors were killed on postoperative day 11, and necropsy was subsequently performed for evaluation of the intestinal adhesions. Results: Significant differences were observed in the control and antibiotic groups (p<0.01) with respect to the survival hours when compared with the saline and E. tirucalli groups. There was no significant difference (p>0.05) in the survival of animals in the saline andE. tirucalli groups; however, one animal died in the saline group. Necropsy of the animals in the saline and E. tirucalligroups showed strong adhesions resistant to manipulation, between the intestinal loops and abdominal wall. The remaining groups did not show any adhesions. Conclusions: Topical treatment with E. tirucalli latex stimulated an increased formation of intestinal adhesions and prevented the death of all animals with peritonitis.

Racional: O uso de plantas da família Euphorbiaceae, principalmente a Euphorbia tirucalli (avelós), tem sido popularmente difundido para o tratamento de uma variedade de doenças de natureza infecciosa, tumoral e inflamatória. Objetivo: Avaliar o efeito do tratamento tópico com a solução aquosa do látex do avelós na sobrevida e nas aderências intestinas de ratos com peritonite experimental. Métodos: Foi induzido peritonite em 24 ratos Wistar e randomizados em quatro grupos de seis, assim distribuídos: 1) Controle - (n=6), nenhum tratamento; 2) Antibiótico - (n=6), tratamento com dose única intramuscular de antibiótico Unasyn (Pfizer - São Paulo); 3) Salina - (n=6), lavagem da cavidade abdominal com solução fisiológica 0,9%; 4) E.Tirucalli - (n=6), lavagem da cavidade abdominal com E. tirucalli na concentração de 12 mg/ml. Os animais que morreram foram submetidos à necropsia e o horário do óbito anotado. Os sobreviventes foram submetidos à eutanásia no 11odia de pós-operatório e, posteriormente, realizou-se a necropsia para avaliação da formação de aderências. Resultados: Os grupos controle e antibiótico obtiveram diferença significativa (p<0,01) com relação às horas de vida entre os grupos salina e E. tirucalli. Não houve diferença significativa (p>0,05) na sobrevida dos animais dos grupos salina e E. tirucalli, no entanto, houve um óbito no grupo salina. A necropsia dos animais dos grupos salina e E. tirucalli mostrou aderências firmes e resistentes à manipulação entre alças intestinais e parede abdominal. Os demais grupos não tiveram formação de aderências. Conclusão: O tratamento tópico com o látex da E. tirucalli estimulou maior formação de aderências intestinais e evitou o óbito de todos animais com peritonite até o período avaliado.

Properties of the tibialis anterior muscle after treatment with laser therapy and natural latex protein following sciatic nerve crush.

INTRODUCTION: In this study we evaluated the characteristics of the tibialis anterior muscle after sciatic nerve crush and treatment with low-level laser therapy (LLLT) or the protein from natural latex (P1). METHODS: We studied the following 6 groups of male Wistar rats: control (CG); exposed nerve (EG); injured nerve (IG); injured nerve with LLLT (LG); injured nerve with P1 (PG); and injured nerve with P1 and LLLT (LPG). RESULTS: After 4 weeks, muscle morphology showed improvement in the treated groups; after 8 weeks, the treated groups resembled controls, especially the PG. Morphometry revealed muscle fiber atrophy after nerve injury, with time-dependent recovery. Histochemical analysis revealed increased intermediate fiber area. The PG was more similar to controls with NADH staining, whereas the LPG more closely resembled controls with SDH staining. CONCLUSION: Treatment using only P1 proved most efficient, revealing a negative interaction between P1 and LLLT.

EFFECTS OF TOPICAL TREATMENT WITH EUPHORBIA TIRUCALLI LATEX ON THE SURVIVAL AND INTESTINAL ADHESIONS IN RATS WITH EXPERIMENTAL PERITONITIS.

BACKGROUND: The use of plants of the family Euphorbiaceae, particularly Euphorbia tirucalli (avelós) has been popularly widespread for treating a variety of diseases of infectious, tumoral, and inflammatory. AIM: To demonstrated antimicrobial and immunomodulatory effects of these extracts, evaluating the effect of a topical treatment with an aqueous solution of avelós latex on the survival and on intestinal adhesions in rats with experimental peritonitis. METHODS: Peritonitis was induced in 24 Wistar rats, that were randomized into four groups of six as follows: (1) Control group (n=6), no treatment; (2) Antibiotic group (n=6), treatment with a single intramuscular dose of antibiotic Unasyn; (3) Saline group (n=6), the abdominal cavity was washed with 0.9% saline; and (4) E.tirucalli group (n=6), the abdominal cavity was washed with E. tirucalli at a concentration of 12 mg/ml. The animals that died were necropsied, and the time of death was recorded. The survivors were killed on postoperative day 11, and necropsy was subsequently performed for evaluation of the intestinal adhesions. RESULTS: Significant differences were observed in the control and antibiotic groups (p<0.01) with respect to the survival hours when compared with the saline and E. tirucalli groups. There was no significant difference (p>0.05) in the survival of animals in the saline andE. tirucalli groups; however, one animal died in the saline group. Necropsy of the animals in the saline and E. tirucalligroups showed strong adhesions resistant to manipulation, between the intestinal loops and abdominal wall. The remaining groups did not show any adhesions. CONCLUSIONS: Topical treatment with E. tirucalli latex stimulated an increased formation of intestinal adhesions and prevented the death of all animals with peritonitis.

Ficus carica latex prevents invasion through induction of let-7d expression in GBM cell lines.

Glioblastoma multiforme (GBM) is one of the deadliest human malignancies. A cure for GBM remains elusive, and the overall survival time is less than 1 year. Thus, the development of more efficient therapeutic approaches for the treatment of these patients is required. Induction of tumor cell death by certain phytochemicals derived from medicinal herbs and dietary plants has become a new frontier for cancer therapy research. Although the cancer suppressive effect of Ficus carica (fig) latex (FCL) has been determined in a few cancer types, the effect of this latex on GBM tumors has not been investigated. Therefore, in the current study, the anti-proliferative activity of FCL and the effect of the FCL-temozolomide (TMZ) combination were tested in the T98G, U-138 MG, and U-87 MG GBM cell lines using the WST-1 assay. The mechanism of cell death was analyzed using Annexin-V/FITC and TUNEL assays, and the effect of FCL on invasion was tested using the chick chorioallantoic membrane assay. To determine the effect of FCL on GBM progression, the expression levels of 40 GBM associated miRNAs were analyzed in T98G cells using RT-qPCR. According to the obtained data, FCL causes cell death in GBM cells with different responses to TMZ, and this effect is synergistically increased in combination with TMZ. In addition, the current study is the first to demonstrate the effect of FCL on modulation of let-7d expression, which may be an important underlying mechanism of the anti-invasive effect of this extract.

Liberação de componentes do extrato de Casearia sylvestris Swartz empregando membranas de látex natural como suporte / Analysis of Casearia sylvestris Swartz extract release from natural rubber latex membranes

A membrana de látex natural (NRL - Natural Rubber Latex), manipulada a partir do látex extraído da seringueira Hevea brasiliensis, tem apresentado grande potencial de aplicação no campo da biomedicina e dos biomateriais. Graças a sua biocompatibilidade e baixa taxa de rejeição, ela tem sido utilizada para próteses e enxertos, atuando como estimulante da angiogênese, adesão celular e barreira física a agentes infecciosos. Além dessas aplicações, as membranas são utilizadas como matriz de sistemas de liberação para avaliar o comportamento da liberação de fármacos e extratos de origem vegetal que apresentam propriedades medicinais. O sistema extrato-membrana tem como objeto de estudo uma nova abordagem dessas substâncias no tratamento de feridas visando à cicatrização e regeneração do tecido envolvido. Casearia sylvestris, conhecida popularmente como guaçatonga, pertence à família Salicaceae, conhecida na medicina popular pelos seus efeitos antiulcerogênicos, cicatrizantes, antiofídicos, anti-inflamatórios e antissépticos, propriedades já comprovadas por estudos. Trabalhos recentes demonstraram que a liberação controlada de fármacos e extratos utilizando membranas de látex natural é uma alternativa interessante e promissora para aplicações biomédicas. Assim, o objetivo deste projeto foi estudar a liberação do extrato de Casearia sylvestris incorporado à membrana de látex em soluções com diferentes valores de pH, com o propósito de estudar seu comportamento e sua liberação de forma controlada. A taxa de liberação do extrato do sistema extrato-membrana foi monitorada e analisada utilizando-se o método de espectroscopia ótica (UV). O pH básico age desestabilizando a membrana indicando que o polímero possui a habilidade de proteger o extrato de ser liberado em valores de pH ácidos, direcionando sua aplicação para o sítio de melhor absorção. Além disso, a liberação segue uma função bi-exponencial...

Natural Rubber Latex (NRL) membrane, made from latex extracted from the rubber tree Hevea brasiliensis, has shown great potential for use in the biomedicine and biomaterials area. Thanks to its biocompatibility and low rejection rate, it has been used for implants and grafts, acting as a stimulant of angiogenesis and cell adhesion and as a barrier against infectious agents. Besides these applications, the membranes are used as model release systems, to assess the release behavior of drugs and plant extracts that exhibit medicinal properties. The extract-membrane system represents a new approach to studying these substances, as aids to wound healing and tissue regeneration. Casearia sylvestris, popularly known as guaçatonga, belongs to the family Salicaceae, known in popular medicine for its anti-ulcerogenic, wound healing, anti-ophidian, antiinflammatory and antiseptic properties, all of which are proven by scientific studies. Recent studies have also shown that the controlled release of drugs and extracts from natural latex membranes is an interesting and promising process for biomedical applications. The objective of this project was thus to study the release of Casearia sylvestris extract incorporated into natural rubber membranes. The main concern was to study and optimize the controlled release of the extract at various pHs. The rate of release was monitored and analyzed by the method of optical spectroscopy (UV). Basic pHacts to destabilize the membrane, indicating that the polymer has the ability to protect the extract from being released at acidic pH values. The controlled release follows a bi-exponential function...

Wound-healing and potential anti-keloidal properties of the latex of Calotropis procera (Aiton) Asclepiadaceae in rabbits.

Calotropis Procera (CP) has been used in the management of toothache, fresh skin burns, gum bleeding as well as others to make it qualify as a medicinal plant. This study was designed to assess its wound-healing property in rabbits and its potentials for anti keloidal activity.Fresh latex of Calotropis were obtained and evaluated phytochemically. Fifteen male rabbits were used and four excisional wounds were created on each rabbit. The rabbits were divided into five groups of three each. Group 1 was the negative control and received no treatment. The wounds of group 2 animals were treated with 2mL of Calotropis latex; group 3 with 2mL honey; and group 4 with a mixture of 1ml honey and 1 mL of the latex. The animals in group 5 were given 2mg triamcinolone intramuscularly. All the groups had their wounds treated daily for 21 days. The wounds' diameters were measured on the day of wound creation, thereafter on days 7, 14 and 21 post wound creation. Biopsies of the wounds were taken on days 3 and 21 and viewed histologically. Phytochemical study of the latex revealed the presence of glycosides, tannins and alkaloids. The wounds were found to be significantly (p<0.05) reduced in groups treated with 50% latex in honey and triamcinolone, respectively, on day 7 post wound creation while there was a significant (p<0.05) reduction in wound surface area in all treated groups on days 14 and 21 post wound creation. Histological findings in untreated group showed thick bundle of collagen fibres some of which had broad based configurations, reminiscent of keloid. The group treated with 2mL of Calotropis latex revealed the presence of florid granulation tissues on day 3 while there was a marked reduction in quantity and size of collagen fibres on day 21 post wound creation which was comparable with what was seen for the triamcinolone-treated group.The general effect of Calotropis latex on wound-healing was noted. Likewise it's similarity to that of triamcinolone, an anti-keloidal agent; this makes it a probable candidate for future anti-keloidal study using a suitable model.

Antitumoural effect of Synadenium grantii Hook f. (Euphorbiaceae) latex.

ETHNOPHARMACOLOGICAL RELEVANCE: Synadenium grantii Hook f. has traditionally been used to treat various neoplastic diseases in southern Brazil. AIM OF STUDY: Evaluation of the antitumoural potential of Synadenium grantii latex against B16F10 melanoma cell line using in vitro and in vivo models, as well as a phytochemical study of the latex. MATERIALS AND METHODS: The in vitro antitumoural activity was performed using MTT and trypan blue assays with different latex concentrations (1.7 µg-7.0 µg/well and 1.22 mg-4.88 mg/well). Flow cytometry was used to determine the progression of the cell cycle. The in vivo activity was performed by subcutaneously injecting melanoma cells in the dorsum of C57BL6 mice, followed by treating the mice with a popular form of use of the latex (garrafada) administered orally. After sacrificing the animals, histological analysis of the organs was performed by hematoxylin-eosin staining. The phytochemical study of the latex was performed by NMR and chromatographic procedures and the extracts and isolated substances were evaluated by IR, 1D and 2D NMR analysis. RESULTS: The Synadenium grantii latex exhibited decreased cell viability of the melanoma line in a concentration and time-dependent manner, and also cell cycle arrest in the S-G2/M phase. The latex caused a 40% reduction in the volume of tumours of the mice with melanomas. Histological examination of the organs of these animals showed no differences between groups. The phytochemical investigation resulted in the isolation and identification of triterpene euphol and the steroid citrostadienol, which were tested against the strain of melanoma. Euphol showed no antitumoural activity, while the steroid citrostadienol showed reduced cytotoxic activity. CONCLUSION: The Synadenium grantii latex presented in vitro and in vivo cytotoxic effects with antitumoural activity against B16F10 melanoma cells.

Protective effect of Calotropis procera latex extracts on experimentally induced gastric ulcers in rat.

AIM OF THE STUDY: Calotropis procera is a wild growing plant with multifarious medicinal properties. The present study was carried out to evaluate the effect of dried latex (DL) of Calotropis procera and its methanol extract (MeDL) against gastric ulcers induced in rats. MATERIALS AND METHODS: Aqueous suspension of DL (20 and 100mg/kg) and MeDL (10 and 50mg/kg) were given orally to 36h fasted rats and ulcers were induced by ethanol, pyloric ligation and aspirin. Parameters like ulcer score and levels of oxidative stress markers were measured in all the models. The effect on gastric hemorrhage and tissue histology was studied in ethanol model and on acidity, pH and volume of gastric secretion was evaluated in pyloric ligation model. The protective effect of DL and MeDL was compared with that of standard anti-ulcer drug famotidine (20 mg/kg). RESULTS: DL and MeDL produced 85-95% inhibition of gastric mucosal damage in ethanol model and 70-80% inhibition in aspirin model. The protective effect of these extracts was associated with marked reduction in gastric hemorrhage, maintenance of tissue integrity and normalization of levels of oxidative stress markers like glutathione, thiobarbituric acid reactive substances and superoxide dismutase. Like famotidine, DL and MeDL decreased the gastric acidity from 376.17+/-21.47 mequiv./l to 163.88+/-6.86 and 201.48+/-8.86 mequiv./l respectively in pyloric ligation model. These extracts increased the gastric pH without affording any protection to gastric mucosa in this model. CONCLUSION: The latex of Calotropis procera has the therapeutic potential to relieve gastric hyperacidity and to prevent gastric ulceration induced by necrotizing agents.

Calotropis procera latex affords protection against carbon tetrachloride induced hepatotoxicity in rats.

In the present study, latex of Calotropis procera possessing potent antioxidant and anti-inflammatory properties was evaluated for its hepatoprotective effect against carbon tetrachloride (CCl(4)) induced hepatotoxicity in rats. Subcutaneous injection of CCl(4,) administered twice a week, produced a marked elevation in the serum levels of aspartate transaminase (AST), alanine transaminase (ALT) and tumor necrosis factor alpha (TNF-alpha). Histological analysis of the liver of these rats revealed marked necro-inflammatory changes that were associated with increase in the levels of TBARS, PGE(2) and catalase and decrease in the levels of glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx). Daily oral administration of aqueous suspension of dried latex (DL) of Calotropis procera at 5, 50 and 100mg/kg doses produced a dose-dependent reduction in the serum levels of liver enzymes and inflammatory mediators and attenuated the necro-inflammatory changes in the liver. The DL treatment also normalized various biochemical parameters of oxidative stress. Our study shows that the antioxidant and anti-inflammatory effects of DL and silymarin were comparable and suggests that DL could be used as a hepatoprotective agent.

[Cytotoxic activity of two cyclic peptides from the latex of Jatropha integerrima Euphorbiaceae]. / Etude de l'activite cytotoxique de deux cyclopeptides isoles du latex de jatropha integerrima euphorbiaceae.

Activity-guided fractionation of the ethyl acetate extract of the latex of Jatropha integerrima Euphorbiaceae combinated with cytotoxic assay against the KB human nasopharyngeal carcinoma cells, resulted to the isolation by chromatographic methods including HPLC of two new cyclopeptides: integerrimacyclopeptide4 and integerrima cyclopeptide B. The amino acid composition was established after hydrolysis and derivatization. All chiral amino acids were L configuration and were hydrophobic. Their sequences were determined by MS fragmentations (ESI-q-TOF, MS/MS) and confirmed by 2D NMR homo- and heteronuclear studies. Integerrimacyclopeptide A was a cyclooctapeptide with m/z 766 corresponding to the molecular formula C37H66N8O9. Analysis of mass spectra gave b(n) and a(n) acylium ion series which the sequence could be deduced: cyclo(Leu-Gly-Ser-Ile-Ile-Leu-Gly-lle). This structure was confirmed by interpretation of HMBC and ROESY spectra. Likewise, integerrimacyclopeptide B was a cycloheptapeptide with m/z 651 and C31H53N7O8 as molecular formula containing one proline residue: cyclo(Pro-Gly-Thr-Ile-Gly-Ile-Leu). These two cyclic peptides exhibited significant cytotoxic activityin vitro against KB tumorales cells with respective IC50 values of 0.46 +/- 0.04 and 0.66 +/-0.08 microg/ml.

Sublingual immunotherapy with a latex extract in paediatric patients: a double-blind, placebo-controlled study.

BACKGROUND: Natural rubber latex (NRL) allergy remains an important allergic disease triggering urticaria, asthma, angioedema and anaphylaxis. Specific immunotherapy can help to solve problems related to NRL allergy. So far, no controlled clinical trials have been performed in children suffering from NRL allergy. OBJECTIVES: To evaluate the safety and efficacy of sublingual immunotherapy (SLIT) with a commercial NRL extract in children with NRL allergy. RESEARCH DESIGN: Randomized, double-blind, placebo-controlled, 12-month trial. PATIENTS AND METHODS: Twenty-six children (aged 4-15 years) with NRL allergy, who had cutaneous and/or respiratory symptoms, including oral allergy syndrome to fruits containing cross-reactive allergens, were recruited. Twelve children were randomized to SLIT with a commercial NRL extract and eight to placebo (3:2). An additional six children with NRL allergy served as untreated controls. MAIN OUTCOMES MEASURES: A glove use test was utilized to monitor skin and systemic symptoms triggered by NRL exposure at baseline and 3, 6, 9 and 12 months later. Oral allergy symptoms were also monitored. RESULTS: No side effects related to treatments were observed in any patient. A significant improvement of symptom score in treated patients in comparison with baseline values was observed at 3 months (p = 0.01) and consolidated after 1 year of treatment (p = 0.0005). In comparison with placebo, significant improvements were observed starting at 9 months from study start (p = 0.015) and at 12 months (p = 0.005). The number of foods triggering oral allergy symptoms increased in placebo and control subjects, but not in active treated patients (p = 0.05). CONCLUSION: Latex SLIT was safe and efficacious in paediatric patients with NRL allergy.

Anticancer and cytotoxic properties of the latex of Calotropis procera in a transgenic mouse model of hepatocellular carcinoma.

AIM: To evaluate the anticancer property of the dried latex (DL) of Calotropis procera, a tropical medicinal plant, in the X15-myc transgenic mouse model of hepatocellular carcinoma and to elucidate its mechanism of action in cell culture. METHODS: The young transgenic mice were orally fed with the aqueous suspension of DL (400 mg/kg for 5 d/wk) for 15 wk and their liver was examined for histopathological changes at 20 wk. Serum levels of vascular endothelial growth factor (VEGF) were also measured in these animals. To characterize the active fraction, DL was extracted with petroleum ether followed by methanol. The methanolic extract was sub-fractionated on a silica gel G column using a combination of non-polar and polar solvents and eleven fractions were obtained. Each fraction was analysed for cytotoxic effect on hepatoma (Huh7) and non-hepatoma (COS-1) cell lines and non-transformed hepatocytes (AML12) using tetrazolium (MTT) assay. Finally, the mechanism of cell death was investigated by measuring the levels of Bcl2, caspase 3 and DNA fragmentation. RESULTS: DL treatment of mice showed a complete protection against hepatocarcinogenesis. No adverse effect was observed in these animals. The serum VEGF level was significantly lowered in the treated mice as compared to control animals. Cell culture studies revealed that the methanolic extract of DL as well as its fraction 8 induced extensive cell death in both Huh-7 and COS-1 cells while AML12 cells were spared. This was accompanied by extensive fragmentation of DNA in Huh-7 and COS-1 cells. No change in the levels of canonical markers of apoptosis such as Bcl2 and caspase 3 was observed. CONCLUSION: DL of C. procera has the potential for anti-cancer therapy due to its differentiable targets and non-interference with regular pathway of apoptosis.