Double impact: natural molluscicide for schistosomiasis vector control also impedes development of Schistosoma mansoni cercariae into adult parasites.

BACKGROUND: Schistosomiasis has been reported in 78 endemic countries and affects 240 million people worldwide. The digenetic parasite Schistosoma mansoni needs fresh water to compete its life cycle. There, it is susceptible to soluble compounds that can affect directly and/or indirectly the parasite's biology. The cercariae stage is one of the key points in which the parasite is vulnerable to different soluble compounds that can significantly alter the parasite's life cycle. Molluscicides are recommended by the World Health Organization for the control of schistosomiasis transmission and Euphorbia milii latex is effective against snails intermediate hosts. METHODOLOGY/PRINCIPAL FINDINGS: We used parasitological tools and electron microscopy to verify the effects of cercariae exposure to natural molluscicide (Euphorbia milii latex) on morphology, physiology and fitness of adult parasite worms. In order to generate insights into key metabolic pathways that lead to the observed phenotypes we used comparative transcriptomics and proteomics. CONCLUSIONS/SIGNIFICANCE: We describe here that the effect of latex on the adult is not due to direct toxicity but it triggers an early change in developmental trajectory and perturbs cell memory, mobility, energy metabolism and other key pathways. We conclude that latex has not only an effect on the vector but applies also long lasting schistosomastatic action. We believe that these results are of interest not only to parasitologists since it shows that natural compounds, presumably without side effects, can have an impact that occurred unexpectedly on developmental processes. Such collateral damage is in this case positive, since it impacts the true target of the treatment campaign. This type of treatment could also provide a rational for the control of other pests. Our results will contribute to enforce the use of E. milii latex in Brazil and other endemic countries as cheap alternative or complement to mass drug treatment with Praziquantel, the only available drug to cure the patients (without preventing re-infection).

Evaluation of brachial plexus fascicles involvement on infraclavicular block: unfixed cadaver study.

BACKGROUND AND OBJECTIVES: This study shows how the diffusion of the anesthetic into the sheath occurs through the axillary infraclavicular space and hence proves the efficacy of the anesthetic block of the brachial plexus, and may thereby allow a consolidation of this pathway, with fewer complications, previously attached to the anesthesia. MATERIALS AND METHODS: 33 armpits of adult cadavers were analyzed and unfixed. We injected a solution of neoprene with latex dye in the infraclavicular space, based on the technique advocated by Gusmão et al., and put the corpses in refrigerators for three weeks. Subsequently, the specimens were thawed and dissected, exposing the axillary sheath along its entire length. RESULTS AND DISCUSSION: Was demonstrated involvement of all fasciculus of the plexus in 51.46%. In partial involvement was 30.30%, 18.24% of cases the acrylic was located outside the auxiliary sheath involving no issue. CONCLUSIONS: The results allow us to establish the infraclavicular as an effective and easy way to access plexus brachial, because the solution involved the fascicles in 81.76% partially or totally, when it was injected inside the axillary sheath. We believe that only the use of this pathway access in practice it may demonstrate the efficiency.

Morphometric and high resolution scanning electron microscopy analysis of low-level laser therapy and latex protein (Hevea brasiliensis) administration following a crush injury of the sciatic nerve in rats.

This study evaluated the effect of low-level laser therapy (LLLT; 15 J/cm(2)) and a latex protein (F1) on a crush injury of the sciatic (ischiadicus) nerve. Seventy-two rats (male, 250 g) were divided into 6 groups: CG, control; EG, exposed nerve; IG, injured nerve without treatment; LG, injured nerve with LLLT; HG, injured nerve with F1; and LHG, injured nerve with LLLT and F1. After 4 or 8 weeks, the animals were euthanized and samples of the sciatic nerve were collected for morphometric and high-resolution scanning electron microscopy (HRSEM) analysis. After 4 weeks, the morphometry revealed improvements in the treated animals, and the HG appeared to be the most similar to the CG; after 8 weeks, the injured groups showed improvements compared to the previous period, and the results of the treatment groups were more similar to one another. At HRSEM after 4 weeks, the treated groups were similar and showed improvement compared to the IG; after 8 weeks, the LHG and HG had the best results. In conclusion, the treatments resulted in improvement after the nerve injury, and this recovery was time-dependent. In addition, the use of the F1 resulted in the best morphometric and ultrastructural findings.

Application of a low-level laser therapy and the purified protein from natural latex (Hevea brasiliensis) in the controlled crush injury of the sciatic nerve of rats: a morphological, quantitative, and ultrastructural study.

This study analyzed the effects of a low-level laser therapy (LLLT, 15 J/cm(2), 780 nm wavelength) and the natural latex protein (P1, 0.1%) in sciatic nerve after crush injury (15 Kgf, axonotmesis) in rats. Sixty rats (male, 250 g) were allocated into the 6 groups (n = 10): CG-control group; EG-nerve exposed; IG-injured nerve without treatment; LG-crushed nerve treated with LLLT; PG-injured nerve treated with P1; and LPG-injured nerve treated with LLLT and P1. After 4 or 8 weeks, the nerve samples were processed for morphological, histological quantification and ultrastructural analysis. After 4 weeks, the myelin density and morphological characteristics improved in groups LG, PG, and LPG compared to IG. After 8 weeks, PG, and LPG were similar to CG and the capillary density was higher in the LG, PG, and LPG. In the ultrastructural analysis the PG and LPG had characteristics that were similar to the CG. The application of LLLT and/or P1 improved the recovery from the nerve crush injury, and in the long term, the P1 protein was the better treatment used, since only the application of LLLT has not reached the same results, and these treatments applied together did not potentiate the recovery.

Anticancer activity of flavonol and flavan-3-ol rich extracts from Croton celtidifolius latex.

CONTEXT: Croton celtidifolius Baill (Euphorbiaceae) is a tree found in the Atlantic Forest in Southern Brazil, where it is commonly known as "Sangue-de-Dragão". Its red latex is used traditionally for treating ulcers, diabetes and cancer. OBJECTIVE: To evaluate antitumor activities of Croton celtififolius latex in vitro and in vivo. MATERIAL AND METHODS: Phytochemical analyses were conducted using HPLC-DAD-MS. Cytotoxic, nuclease and pro-apoptotic properties were determined using the tetrazolium salt assay (MTT), plasmid DNA damage assay and ethidium bromide (EB)/acridine orange methods, respectively, and antitumor activity was determined in the Ehrlich ascites carcinoma (EAC) mouse model. RESULTS: Phytochemical studies indicated a high phenol content of flavonols (45.67 ± 0.24 and 18.01 ± 0.23 mg/mL of myricetin and quercetin, respectively) and flavan-3-ols (114.12 ± 1.84 and 1527.41 ± 16.42 mg/L of epicatechin and epigallocatechin, respectively) in latex. These compounds reduced MCF-7 and EAC cell viability in the MTT assay (IC50 = 169.0 ± 1.8 and 187.0 ± 2.2 µg/mL, respectively). Latex compounds caused significant DNA fragmentation and increased the number of apoptotic cells (negative control (NC), 12%; latex, 41%) as indicated by differential staining in the EB/acridine orange assay. The in vivo latex treatment at 3.12 mg/kg/day reduced the body weight by 7.57 ± 2.04 g and increased median survival time to 17.5 days when compared to the NC group (13.0 days). In addition, the highest latex concentration inhibited tumor growth by 56%. DISCUSSION AND CONCLUSION: These results agree with ethno-pharmacological reports showing cytotoxicity and antitumor activity of C. celtidifolius latex. The mechanism of antitumor action may be related to direct DNA fragmentation that reduces survival and induces apoptosis.

Avaliação da influência do Látex aquoso de Euphoria milli var. hislopii (EUPHORBIACEAE) sobre o ciclo biológico de Schistosoma mansoni (TREMATODA, SCHISTOSOMATIDAE) em condições laboratoriais

A esquistossomose está entre as doenças parasitárias mais prevalentes do mundo. Diante da problemática do controle da infecção esquistossomótica, a Organização Mundial de Saúde (OMS) tem estimulado o desenvolvimento de estratégias que objetivam a redução/eliminação do ciclo de Schistosoma mansoni em áreas endêmicas. O extrato aquoso do látex de Euphorbia milli (syn. slpendens) var. hislopii apresenta relevantes características como produto seletivo no controle da endemia em larga escala. A presença das formas evolutivas no ambiente (ovos, miracídios e cercarias) e os períodos pré-patente e patente no hospedeiro intermediário, são fases críticas do desenvolvimento do ciclo, dado que nesses períodos tais formas encontram-se mais suceptíveis a ação de produtos veiculados no recurso hídrico. A presente dissertação apresentará o formato de artigo e será dividida em três capítulos, sendo eles: 1- Efeito do moluscidia E. milli var. hislopii no desenvolvimento de S. mansoni em Biomphalaria glabrata,2- Controle da dinâmica parasitária de S. mansoni em B. glabrata através do tratamento de ovos e miracídios com E. milli var. hislopii e, 3- Influência do látex de E. milli var. hislopii na formação de S. mansoni no hospedeiro definitivo. Com base nos resultados obtidos, podemos concluir que a propriedade moluscicida e esquistossomicida do produto em questão, viabilizada pela ação do mesmo nas formas infectantes e nos hospedeiros intermediários promove de forma concomitante e eficiente o controle da transmissão e da morbidade da doença. Um único produto, natural, biodegradável, em baixas doses, viável ecologicamente altera o desenvolvimento do ciclo do parasito nos hospedeiros intermediários e definitivos, gerando a redução de formas infectantes e a preservação das populações de moluscos em seu habitat natural

Rabbit rubeosis iridis induced by intravitreal latex-derived angiogenic fraction.

PURPOSE: To describe the presence of iris neovascularization in a rabbit-model of retinal neovascularization induced by the intravitreal injection of latex-derived angiogenic fraction microspheres (LAF). MATERIALS AND METHODS: Eight New Zealand rabbits received one intravitreal injection of PLGA (L-lactide-co-glycolide) microspheres with 50 ug of LAF in the right eye (Group A). Microspheres without the LAF (0.1 ml) were injected in controls (Group B; n = 8). Follow-up with clinical evaluation and iris fluorescein angiography was performed after 4 weeks when eyes were processed for light microscopy. RESULTS: All eyes from Group A showed significant vascular dilation, conjunctival hyperemia and neovascularization on the iris surface, after LAF injection. No vascular changes were observed in Group B. CONCLUSIONS: The intravitreal injection of microspheres containing the LAF can induce rubeosis iridis in rabbits and could be used as a simple experimental model for iris neovascularization.

Bioactivity of latex from Euphorbia splendens var. hislopii (Euphorbiaceae) on post-embryonic development of Megaselia scalaris (Phoridae).

Larvae of Megaselia scalaris (Loew, 1866) feed on a wide range of decomposing organic matter and present a great importance to public health. This study evaluated the effect of crude latex extract from Euphorbia splendens var. hislopii (Euphorbiaceae) on post-embryonic development time of M. scalaris under laboratory conditions. The latex was used in its crude lyophilized form, dissolved in distilled water and tested in concentrations of 5 microg/mL, 10 microg/mL and 20 microg/mL. The latex was applied with the aid of an automatic pipette (1 microL/larva) on the newly-hatched larvae. Each group (the three concentrations of latex and the control group) was composed of 50 larvae and fed with 25mg of decomposing horse flesh. The experiment was made in quadruplicate. The observations were recorded daily. The data were submitted to analysis of variance (ANOVA) and Tukey's post hoc-test with a 5% significance level. The post-embryonic development time for all stages (larval, pupal and newly-hatched larvae to adult) tested with all three latex concentrations was significantly shorter than for the control group, but without any significant difference among the different concentrations. The more sensitive stages to the substance were pupal and newly-hatched larvae to adult. The viability was less than 51.5% in the three concentrations of latex in these stages and they were lower than for the control group (67.4% for pupal stage and 64% for newly-hatched larvae to adult). Therefore, it is likely that this substance has influence on the development and viability of these flies and can become a promising agent for insect pest management.

Leishmanicidal activity of Himatanthus sucuuba latex against Leishmania amazonensis.

Himatanthus sucuuba (HsL) latex exhibited a potent leishmanicidal activity against intracellular amastigotes of Leishmania amazonensis, a causative agent of cutaneous leishmaniasis. HsL inhibited intracellular amastigote growth in a dose-dependent manner (IC(50)=15.7microg/mL). Moreover, HsL increased nitric oxide (NO) and Tumor Nuclear Factor-alpha (TNF-alpha) and decreased Transforming Growth Factor-beta (TGF-beta) production in macrophages. As assessed by plasma membrane integrity and mitochondrial activity, HsL showed low toxicity for host macrophages. HsL in vivo was active by the oral route, reducing the parasite load in established footpad lesions after only five doses. In summary, these findings support HsL as an interesting candidate for further evaluations regarding its potential application as a therapeutical agent against Leishmania.

Immunological and allergenic responses induced by latex fractions of Calotropis procera (Ait.) R.Br.

Immunological and allergenic responses against the latex of Calotropis procera were investigated in mice by oral and subcutaneous routes. The latex was fractionated according to water solubility and molecular size of its components. The fractions were named as non-dialyzable latex (NDL) corresponding to the major latex proteins, dialyzable latex (DL) corresponding to low molecular size substances and rubber latex (RL) which was highly insoluble in water. Anti-sera against these fractions were assayed for total IgG and IgA titration by ELISA and IgE and IgG(1) were quantified by passive cutaneous anaphylaxis (PCA) in rats and mice, respectively. None of the fractions induced antibodies level increases when mice received latex fractions by oral route and thus, did not develop allergy. Nonetheless, anti-sera of mice sensitized with NDL and RL by subcutaneous route displayed considerable immunological response while DL did not. IgG level augmented consistently against NDL and RL while IgA response was detected only to NDL. NDL and RL induced very strong PCA reactions suggesting that both fractions would contain latex substances involved in allergy. Furthermore, protein analysis of NDL and RL suggests that RL still retain residual proteins abundantly found in NDL that could explain its similar allergenic effect. No IgG(1) reaction was detected in any of the anti-sera tested. According to the results, the proteins of latex of Calotropis procera can provoke allergy by subcutaneous route. The NDL has previously shown to display anti-inflammatory and analgesic activities by intraperitoneal injection. It should be relevant to determine whether NDL could induce such activities when assayed by oral route since it was ineffective to induce allergy by this way.

Biocompatibility of natural latex implanted into dental alveolus of rats.

The present study investigated the biocompatibility of a biopolymer based on vegetable latex extracted from the Hevea brasiliensis rubber tree, implanted into the bony alveolar cavity after dental extraction in rats. A granule of latex (area = 0.25 +/- 0.04 mm(2)) was implanted inside the alveolus immediately after extraction of the upper right incisor, and the animals were sacrificed 7, 21 and 42 days after the procedure. The hemi-maxillas were decalcified and processed for embedding in paraffin to obtain semi-serial longitudinal sections 5 mum thick, and then stained with hematoxylin-eosin. The latex granule was observed in the cervical third of the alveolus without any foreign body reaction, or persistence of the initial acute inflammatory reaction. Bone repair in the areas adjacent to the material was quantified, and a decrease was noted in the thickness of the fibrous capsule surrounding the implants from 92.8 +/- 9.3 microm on day 7 to 9.4 +/- 1.8 microm on day 42 (ANOVA, P = 0.01). The quantitative data confirmed acceleration of bone formation (statistically significant at 5%) in parallel with a decrease of connective tissue in the areas around the implants. These results show that the tested material is biologically compatible, and progressively integrated into the alveolar bone, simultaneously accelerating bone formation and playing an important role in the healing process.

Evaluation of the molluscicidal properties of Euphorbia splendens var. hislopii (N.E.B.) latex: experimental test in an endemic area in the State of Minas Gerais, Brazil.

Following the positive results obtained regarding the molluscicidal properties of the latex of Euphorbia splendens that were corroborated in laboratory and field tests under restricted conditions, a field study was conducted in experimental streams located in an endemic area. After recording the average annual fluctuations of vectors in three streams, a solution of E. splendens latex at 12 ppm was applied in stream A, a solution of niclosamide at 3 ppm that was applied in stream B and a third stream (C) remained untreated for negative control. Applications of E. splendens and niclosamide resulted in a mortality of 100% among the snails collected in the streams A and B. No dead snails were found in the negative control stream. A monthly follow-up survey conducted during three consecutive months confirmed the return of vectors to both experimental streams treated with latex and niclosamide. This fact has called for a need to repeat application in order to reach the snails that remained buried in the mud substrate or escaped to the water edge, as well as, newly hatched snails that did not respond to the concentration of these molluscicides. Adults snails collected a month following treatment led us to believe that they had migrate from untreated areas of the streams to those previously treated.

The allergenic properties of fresh and preserved Hevea brasiliensis latex protein preparations.

The allergenic properties of the proteins of two lyophilized fractions of fresh natural rubber latex obtained by ultracentrifugation, the C serum and the sedimented bottom or lutoid fraction, have been compared with those of the serum proteins of two samples of high ammonia latex (HAL) [A]HALS obtained from HAL stored for more than 1 year, and [M]HALS derived from HAL stored for 6 weeks before ultracentrifugation and lyophilization. The most potent source of allergenic polypeptides both for skin prick testing of latex-sensitive patients and for immunoblots of their blood serum was the lutoid fraction of fresh latex. Skin prick tests and immunoblots of patients' sera showed that the allergenicity of the ammoniated latex decreased during storage. Skin prick tests using fractions of [A]HALS, C serum and lutoid proteins obtained after passage through a Sephacryl S300 column showed that the components of all three preparations which eluted in the largest volumes were almost equally effective in provoking the largest number of responses. Immunoblots of the sera of 43 latex-sensitive individuals showed that the majority (66%) of sera of the adult allergic patients reacted with a polypeptide of 19 kD. No characteristic pattern of binding latex polypeptides could be recognized in the sera from patients who were also asthmatic or from those who had an anaphylactic response to latex proteins.

[Cicatrization effect of Jatropha curcas latex (Angiospermae: Euforbiaceae)]. / Actividad cicatrizante del látex de Jatropha curcas (Angiospermae: Euforbiaceae).

The wound healing effects of Jathopha curcas latex upon surgical wound produced in Balb/c mice skin, were studied with a modification of the Hoowes-Sooy-Harvey method. The effects of topical treatment using single 50 ul doses of latex at different dilutions (10% to 100%) was compared with a multiple dose treatment (four 25 ul/dose q12h, latex 5% to 100%). The single dose treatment with 10%, 50% or 100% latex and the multiple dose treatment with dilutions between 5% and 10%, have a healing effect but only on males. The multiple dose treatment with 50% or pure undiluted latex produced caustic lesions to treated skin.