Toxicity of Senna occidentalis seeds in laying hens and its effects on egg production.

Senna occidentalis is a toxic leguminous plant found in many tropical and subtropical regions of the world and causes poisoning mainly in confined animals. The seeds are the most toxic part of the plant and may be present in animal rations. The main toxic component of the S. occidentalis seed is a dianthrone, an anthraquinone-derived compound that affects mitochondrial function. This study evaluated the effects on egg production of low-level contamination of the S. occidentalis in the laying hens' diet. Forty-eight one-day-old pullets were randomly allocated into two treatment groups: control, birds that received no experimental treatment; and external and internal tegument (ET/IT), birds that received a diet containing 0.2% of ET/IT of S. occidentalis seeds throughout their life cycle (42 weeks). The birds were monitored for clinical signs of poisoning, and the production and quality of eggs were recorded. Necropsies were conducted at the end of the experimental period. None of the layers showed any clinical signs of poisoning, decreases in feed intake or alterations of the body weight gain. A marked reduction in egg production and, consequently, a lower feed efficiency in ET/IT group were measured. Ovaries were the most affected organ in birds from the ET/IT group, and yolk leaking and dysplasia of the inner layer of the vitelline membrane were observed. S. occidentalis was shown to be toxic for laying hens. Considering these results, it is feasible to assume that the constant presence of low concentrations of S. occidentalis seeds in rations represents a threat to the poultry industry.

Pfaffosidic Fraction from Hebanthe paniculata Induces Cell Cycle Arrest and Caspase-3-Induced Apoptosis in HepG2 Cells.

Hebanthe paniculata roots (formerly Pfaffia paniculata and popularly known as Brazilian ginseng) show antineoplastic, chemopreventive, and antiproliferative properties. Functional properties of these roots and their extracts are usually attributed to the pfaffosidic fraction, which is composed mainly by pfaffosides A-F. However, the therapeutic potential of this fraction in cancer cells is not yet entirely understood. This study aimed to analyze the antitumoral effects of the purified pfaffosidic fraction or saponinic fraction on the human hepatocellular carcinoma HepG2 cell line. Cellular viability, proliferation, and apoptosis were evaluated, respectively, by MTT assay, BrdU incorporation, activated caspase-3 immunocytochemistry, and DNA fragmentation assay. Cell cycle was analyzed by flow cytometry and the cell cycle-related proteins were analyzed by quantitative PCR and Western blot. The cells exposed to pfaffosidic fraction had reduced viability and cellular growth, induced G2/M at 48 h or S at 72 h arrest, and increased sub-G1 cell population via cyclin E downregulation, p27(KIP1) overexpression, and caspase-3-induced apoptosis, without affecting the DNA integrity. Antitumoral effects of pfaffosidic fraction from H. paniculata in HepG2 cells originated by multimechanisms of action might be associated with cell cycle arrest in the S phase, by CDK2 and cyclin E downregulation and p27(KIP1) overexpression, besides induction of apoptosis through caspase-3 activation.

Immunosuppressive effects of Pteridium aquilinum enhance susceptibility to urethane-induced lung carcinogenesis.

Pteridium aquilinum (bracken fern), one of the most important toxic plants in the world, contains the toxic norsequiterpene ptaquiloside that induces cancers in humans and farm animals. Previous studies in the laboratory demonstrated immunotoxic effects produced by ptaquiloside, which are characterized by suppression of natural killer (NK) cell activity (i.e. cytotoxicity and interferon [IFN]-γ production). However, it is unknown whether these immunosuppressive effects could contribute to carcinogenesis in situ in general because of the important function of NK cells in innate killing of tumor cells. This study assessed the impact of P. aquilinum-induced immunosuppression on urethane-induced lung cancer in C57BL/6 mice. Adult mice were treated with an extract of P. aquilinum (30 g/kg/day) by gavage once daily for 14 days, followed by gavage (5 days/week) during an 11-week period that was accompanied by treatment with urethane (1 g/kg) via once-weekly intraperitoneal injection; 20 weeks after the end of the treatment period, all lungs were evaluated. The results indicated there was a significant increase in lung nodule number as well as in multiplicity of lesions in mice treated with both P. aquilinum and urethane (PU group) compared to values in mice treated only with the urethane (U group). In addition, histologic evaluation revealed a 76% increase in the rate of lung adenomas and a 41% increase in rate of bronchiolization of alveoli in the mice from the PU group compared to levels seen in mice within the U group. Taken together, the results here show for the first time that immunosuppressive effects of P. aquilinum could increase the risk of cancer formation in exposed hosts.

Ptaquiloside reduces NK cell activities by enhancing metallothionein expression, which is prevented by selenium.

Pteridium aquilinum, one of the most important poisonous plants in the world, is known to be carcinogenic to animals and humans. Moreover, our previous studies showed that the immunosuppressive effects of ptaquiloside, its main toxic agent, were prevented by selenium in mouse natural killer (NK) cells. We also verified that this immunosuppression facilitated development of cancer. Here, we performed gene expression microarray analysis in splenic NK cells from mice treated for 14 days with ptaquiloside (5.3 mg/kg) and/or selenium (1.3 mg/kg) to identify gene transcripts altered by ptaquiloside that could be linked to the immunosuppression and that would be prevented by selenium. Transcriptome analysis of ptaquiloside samples revealed that 872 transcripts were expressed differentially (fold change>2 and p<0.05), including 77 up-regulated and 795 down-regulated transcripts. Gene ontology analysis mapped these up-regulated transcripts to three main biological processes (cellular ion homeostasis, negative regulation of apoptosis and regulation of transcription). Considering the immunosuppressive effect of ptaquiloside, we hypothesized that two genes involved in cellular ion homeostasis, metallothionein 1 (Mt1) and metallothionein 2 (Mt2), could be implicated because Mt1 and Mt2 are responsible for zinc homeostasis, and a reduction of free intracellular zinc impairs NK functions. We confirm these hypotheses and show increased expression of metallothionein in splenic NK cells and reduction in free intracellular zinc following treatment with ptaquiloside that were completely prevented by selenium co-treatment. These findings could help avoid the higher susceptibility to cancer that is induced by P. aquilinum-mediated immunosuppressive effects.

Mimosine and cyclophosphamide: a potential new combination therapy used to prevent tumor development

The effects of mimosine (MI), which is an amino acid that is derived from Leucaena leucocephala, were evaluated on the growth of ascitic Ehrlich tumors, and the effects of the combination treatment of MI and cyclophosphamide (CY) on tumor growth were also assessed. Mice were divided into groups that received the following treatments over the course of 20 days: phosphate buffer solution (CO), MI, Ehrlich cells (E), E plus CY (EC), E plus MI (EM) and E plus MI and CY (EMC). No signs of toxicity were detected in the mice from the MI group. The mice from the EMC group showed reductions in body weights when compared with those from the E group. The animals from the EC, EM and EMC groups showed reductions in ascitic volume compared with those from the E group. The mice from the EMC group showed reductions in total cell numbers of ascitic fluid compared with those from the E, EC and EM groups. The combination of MI and CY was the most effective treatment for Ehrlich tumor ascites.

Haematological and immunological effects of repeated dose exposure of rats to integerrimine N-oxide from Senecio brasiliensis.

This study is the first in the literature to focus attention on the possible immunotoxic effect of integerrimine N-oxide content in the butanolic residue (BR) of Senecio brasiliensis, a poisonous hepatotoxic plant that contains pyrrolizidine alkaloids (PAs). PAs have been reported as a pasture and food contaminant and as herbal medicine used worldwide and are responsible for poisoning events in livestock and human beings. After the plant extraction, BR extracted from Senecio brasiliensis was found to contain approximately 70% integerrimine N-oxide by elemental and spectral analyses ((1)H and (13)C NMR), which was administered to adult male Wistar Hannover rats at doses of 3, 6 and 9 mg/kg for 28 days. Body weight gain, food consumption, lymphoid organs, neutrophil analysis, humoural immune response, cellular immune response and lymphocyte analysis were evaluated. Our study showed that integerrimine N-oxide could promote an impairment in the body weight gain, interference with blood cell counts and a reducing T cell proliferative activity in rats; however, no differences in the neutrophil activities, lymphocytes phenotyping and humoural and cellular immune responses were observed. It is concluded that doses of integerrimine N-oxide here employed did not produce marked immunotoxic effects.

Biotechnology tools in agriculture: recent patents involving soybean, corn and sugarcane.

The technological opportunities opened up by biotechnology in agriculture are diverse, including plant breeding, the partial or total relief of pesticides chemicals usage, the improvement of soil fertility, the improvement of the quality attributes of various foods. Specifically, various tricks of biotechnology can be used for higher seed yield, resistance to diseases and insects, better stems and roots, tolerance to drought and heat, and better agronomic quality. A number of recent works considerably widen the potential of plant biotechnology where transformation methods and studies of molecular genomics have been described. For example, transformation techniques and search for new selectable markers involving biolistic technique, gene transfer technique using the soil bacterium Agrobacterium tumefaciens, selection technique based on the use of mannose, utilization of genes promoting endogenous hormone production under the control of chemical stimulants, further more, engineering the nuclear genome without antibiotic resistance genes and engineering the plastid genome. We are presenting in this paper some of the recent patents on methods and techniques involving genes coding proteins and breeding techniques with possible agronomic applicability on crops economically important, such as soybean, corn and sugarcane.

Pharmaceutical properties of 'sucupira' (Pterodon spp. ) / Propriedades farmacêuticas de "sucupira" (Pterodon spp. )

The plant of the genus Pterodon (Fabaceae, Leguminosae), commonly known as 'sucupira' or 'faveira', are disseminated throughout the central region of Brazil and has frequently been used in popular medicine for its anti-rheumatic, analgesic, and anti-inflammatory properties. In recent years, interest in these plants has increased considerably. The biological effects of different phytoextracts and pure metabolites have been investigated in several experimental models in vivo and in vitro. The literature describes flavonoids, triterpene and steroids, while one paper presented studies with proteins isolated from the genus. This review provides an overview of phytochemical and pharmacological research in Pterodon, showing the main chemical compounds studied to date, and focusing on the relationship between these molecules and their biological activity. Furthermore, this study paves the way for more in-depth investigation, isolation and characterization of the molecules of this plant genus.

As plantas do gênero Pterodon (Fabaceae/Leguminosae), conhecidas popularmente como "sucupira branca" ou "faveira", encontram-se distribuídas pela região central do Brasil e são frequentemente utilizadas na medicina popular por suas propriedades antirreumáticas, analgésicas e antiinflamatórias. Nos últimos anos, o interesse por estas plantas tem aumentado consideravelmente. Os efeitos biológicos dos diferentes fitoextratos e metabólitos puros têm sido investigados em vários modelos experimentais in vivo e in vitro. A literatura descreve flavonóides, triterpenos, esteróides e apenas um trabalho mostra estudos com proteínas isoladas do gênero. Esta revisão apresenta de maneira geral as investigações farmacológicas e fitoquímicas de Pterodon, mostrando os principais compostos já estudados, sua composição química, focando na relação entre estas moléculas e sua atividade biológica. Mais ainda, nós abrimos as portas para maior investigação, isolamento e caracterização de moléculas deste gênero de plantas.

Inhibition of ascitic ehrlich tumor cell growth by intraperitoneal injection of Pfaffia paniculata (Brazilian ginseng) butanolic residue

This study aimed to investigate the effects of the administration of butanolic residue (BR) of Pfaffia paniculata by intraperitoneal route to Ehrlich ascitis tumor bearing mice. Initially, a toxicity study of P. paniculata BR was performed in which doses of 12.5; 25 and 50mg/Kg were administered by intraperitoneal injection for seven days to Swiss mice. The treatment did not show toxicity. Then, Swiss male mice received, by intraperitoneal injection, once a day, 12.5; 25 or 50mg/Kg of P. paniculata BR for seven days. This protocol started in the same day of tumor inoculation with 5X10(6) cells i.p. The treatment with butanolic residue of P.paniculata i.p caused a significant increase in the ascitic volume; however, a significant decrease in tumor cells number per ml (p<0.05) was observed in P. paniculata treated mice that was followed by a numerical (although non-significant) decrease in the total numbers of tumor cells in the collected ascitic fluid. These results indicated a tumor cell inhibitory effect by P. paniculata butanolic residue in this experimental system, and indicate that topical application of this residue can be useful to control the cancer growth.

Neste estudo, foi avaliado o efeito do tratamento intraperitoneal com Resíduo Butanólico de Pfaffia paniculata, sobre o crescimento do Tumor de Ehrlich, forma ascítica. Foram utilizados dois grupos de 15 camundongos cada, sendo um grupo controle e o outro grupo tratado com RB 50mg/Kg. Todos os animais foram inoculados intraperitonealmente, com 5X10(6) células tumorais O tratamento iniciou-se no mesmo dia da inoculação do tumor. Assim, os animais receberam diariamente, por via intraperitoneal, 0,1 ml de RB na concentrações 50 mg/Kg, ou PBS como controle. Após 7 dias da inoculação do tumor, os animais foram eutanasiados e foi colhido o fluído ascítico total, para a contagem do número de células tumorais presentes neste fluído e estudo da morfologia destas células . Neste experimento observou-se aumento significante da quantidade de fluido ascítico nos animais tratados com RB, e diminuição significativa em relação ao número de células tumorais/ml e células tumorais totais, presentes no fluído ascítico, comparativamente com os animais controle. Estes resultados sugerem efeito inibitório tópico do RB levando à morte as células neoplásicas.

Pfaffia paniculata (Brazilian ginseng) roots decrease proliferation and increase apoptosis but do not affect cell communication in murine hepatocarcinogenesis.

Pfaffia paniculata (Brazilian ginseng) roots and/or its extracts have shown anti-neoplastic, chemopreventive, and anti-angiogenic properties. The aim of this work was to investigate the chemopreventive mechanisms of this root in mice submitted to the infant model of hepatocarcinogenesis, evaluating the effects on cellular proliferation, apoptosis, and intercellular communication. Fifteen-day-old BALB/c male mice were given, i.p., 10mug/g of the carcinogen N-nitrosodiethylamine (DEN). Animals were separated into three groups at weaning and were given different concentrations of powdered P. paniculata root (0%, 2%, or 10%) added to commercial food for 27 weeks. Control group (CT) was not exposed to the carcinogen and was given ration without the root. After euthanasia, the animals' liver and body weight were measured. Liver fragments were sampled to study intercellular communication, molecular biology, and histopathological analysis. Cellular proliferation was evaluated by immunohistochemistry for PCNA, apoptosis was evaluated by apoptotic bodies count and alkaline comet technique, and intercellular communication by diffusion of lucifer yellow dye, immunofluorescence, western blot and real-time PCR for connexins 26 and 32. Chronic treatment with powdered P. paniculata root reduced cellular proliferation and increased apoptosis in the 2% group. Animals in the 10% group had an increase in apoptosis with chronic inflammatory process. Intercellular communication showed no alterations in any of the groups analyzed. These results indicate that chemopreventive effects of P. paniculata are related to the control of cellular proliferation and apoptosis, but not to cell communication and/or connexin expression, and are directly influenced by the root concentration.

Immunomodulatory effects of Pteridium aquilinum on natural killer cell activity and select aspects of the cellular immune response of mice.

Pteridium aquilinum (bracken fern) is one of the most common plants. Epidemiological studies have revealed a higher risk of certain types of cancers (i.e., esophageal, gastric) in people who consume bracken fern directly (as crosiers or rhizomes) or indirectly through the consumption of milk from livestock that fed on the plant. In animals, evidence exists regarding the associations between chronic bracken fern intoxication, papilloma virus infection, and the development of carcinomas. While it is possible that some carcinogens in bracken fern could be responsible for these cancers in both humans and animals, it is equally plausible that the observed increases in cancers could be related to induction of an overall immunosuppression by the plant/its various constituents. Under the latter scenario, normal tumor surveillance responses against nascent (non-bracken-induced) cancers or responses against viral infections (specifically those linked to induction of cancers) might be adversely impacted by continuous dietary exposure to this plant. Therefore, the overall objective of this study was to evaluate the immunomodulatory effects of bracken fern following daily ingestion of its extract by a murine host over a period of 14 (or up to 30) days. In C57BL/6 mice administered (by gavage) the extract, histological analyses revealed a significant reduction in splenic white pulp area. Among a variety of immune response parameters/functions assessed in these hosts and isolated cells, both delayed-type hypersensitivity (DTH) analysis and evaluation of IFNgamma production by NK cells during T(H)1 priming were also reduced. Lastly, the innate response in these hosts-assessed by analysis of NK cell cytotoxic functionality-was also diminished. The results here clearly showed the immunosuppressive effects of P. aquilinum and that many of the functions that were modulated could contribute to the increased risk of cancer formation in exposed hosts.

Low doses of monocrotaline in rats cause diminished bone marrow cellularity and compromised nitric oxide production by peritoneal macrophages.

Monocrotaline (MCT) is a pyrrolizidine alkaloid found in a variety of plants. The main symptoms of MCT toxicosis in livestock are related to hepato- and nephrotoxicity; in rodents and humans, the induction of a pulmonary hypertensive state that progresses to cor pulmonale has received much attention. Although studies have shown that MCT can cause effects on cellular functions that would be critical to those of lymphocytes/macrophages during a normal immune response, no immunotoxicological study on MCT have yet to ever be performed. Thus, the aim of the present study was to evaluate the effect of MCT on different branches of the immune system using the rat--which is known to be sensitive to the effects of MCT--as the model. Rats were treated once a day by gavage with 0.0, 0.3, 1.0, 3.0, or 5.0 mg MCT/kg for 14 days, and then any effects of the alkaloid on lymphoid organs, acquired immune responses, and macrophage activity were evaluated. No alterations in the relative weight of lymphoid organs were observed; however, diminished bone marrow cellularity in rats treated with the alkaloid was observed. MCT did not affect humoral or cellular immune responses. When macrophages were evaluated, treatments with MCT caused no significant alterations in phagocytic function or in hydrogen peroxide (H2O2) production; however, the MCT did cause compromised nitric oxide (NO) release by these cells.

Steroidal saponin concentrations in Brachiaria decumbens and B. brizantha at different developmental stages / Concentrações de saponina esteroidal em diferentes fases do desenvolvimento de Brachiaria decumbens e B. brizantha

Brachiaria species contain steroidal saponins and are involved in outbreaks of hepatogenous photosensitization. This research presents the levels of a steroidal saponin, protodioscin, in the seeds and aerial parts of B. brizantha and B. decumbens during different developmental stages (growth, bloom, fructification and seed fall). The butanolic fraction of the ethanolic extract of each stage was submitted to thin layer chromatography (TLC) and spectrophotometric analysis through the Ehrlich reagent in 515nm. The chromatograms in TLC of the butanolic fraction of B. brizantha and B. decumbens showed similar spots as the protodioscin standard. The estimated level of protodioscin isomers in B. brizantha and B. decumbens ranged from 0.5 percent to 2.1 percent, having the highest level at the end of their developmental stages during seed falling comparison with the previous one. Protodioscin was not detected in the seeds. Outbreaks of Brachiaria spp. poisoning in central Western Brazil are frequently observed in pastures that had been more than 30 days without animals grazing, and also during the growing or blooming stage of the pastures. Other saponin determinations in toxic and non toxic pastures are necessary to determine the saponin concentrations that cause intoxication.

As Brachiaria spp contêm saponinas esteroidais envolvidas no desenvolvimento de fotossensibilizacão hepatógena. No presente trabalho foram determinados os teores da saponina esteroidal protodioscina nas partes aéreas de B. brizantha e B. decumbens, durante as diferentes fases do desenvolvimento (crescimento, floração, frutificação e queda das sementes) e nas sementes. A fração butanólica do extrato etanólico de cada fase foi submetida à cromatografia de camada delgada (CCD) e à análise espectrofotométrica por meio do reagente de Ehrlich em 515nm. Os cromatogramas em CCD da fração butanólica de B. brizantha e B. decumbens, de cada fase do desenvolvimento, apresentaram manchas similares ao padrão de protodioscina. Por meio da análise de protodioscina por espectrofotometria, os teores de protodioscina em B. brizantha e B. decumbens variaram entre 0.5 por cento e 2.1 por cento, sendo mais altos na fase de queda das sementes. Nas sementes não foi encontrada protodioscina. Surtos de intoxicação por Brachiaria spp. no Centro-Oeste brasileiro são freqüentemente observados em pastagens diferidas por mais de 30 dias e também durante as fases de crescimento e florescimento. São necessárias outras dosagens de saponinas em pastagens tóxicas e não-tóxicas para determinar as concentrações capazes de causar intoxicação.

Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil: [revision] / Intoxicação por Panicum dicotomiflorum em ovinos na região nordeste: [revisão]

Different species of Panicum, including P. dichotomiflorum,have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to ÕH and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

Diferentes espécies de Panicum, incluindo P. dichotomiflorum,causam fotossensibilização em ovinos, eqüinos, bovinos e caprinos. Um surto de fotossensibilização hepatógena ocorreu em 3 rebanhos de ovinos no município de Casa Nova, Bahia. Oitenta e um de 365 ovinos adoeceram e 39 morreram. Cordeiros lactentes e ovinos de menos de um ano foram mais afetados. Jumentos, cabras e bovinos que permaneciam nas mesmas pastagens não foram afetados. Clinicamente observou-se edema da face, seguido por dermatite, principalmente da face, orelhas e garupa, corrimento ocular, opacidade da córnea com cegueira, e avermelhamento do rodete coronário e casco. Na necropsia de um cordeiro afetado o fígado estava amarelado. Na histologia observou-se necrose aleatória de hepatócitos no fígado e áreas focais de necrose de miócitos no miocárdio. Amostras de P. dicotomiflorum foram analisadas por cromatografia em camada delgada e aquelas contendo saponinas foram isoladas por HPLC utilizando uma coluna RP-C18 com mistura de metanol e água. Os compostos isolados foram submetidos a espectroscopia de ressonância magnética nuclear de hidrogênio (RMN ÕH) e de carbono (RMN 13C). As reações foram positivas para saponinas furostanólicas com o mesmo Rf que os padrões de protodioscina (0,21) e metilprotodioscina (0,32). Este resultado e os dados espectroscópicos sugerem a presença, em P. dichotomiflorum, de metilprotodioscina e uma mistura de isômeros (25R)- e (25S)- de protodioscina em uma proporção de 3:1.

Cytotoxic effects of butanolic extract from Pfaffia paniculata (Brazilian ginseng) on cultured human breast cancer cell line MCF-7.

Roots of Pfaffia paniculata have been well documented for multifarious therapeutic values and have also been used for cancer therapy in folk medicine. This study has been performed in a human breast tumor cell line, the MCF-7 cells. These are the most commonly used model of estrogen-positive breast cancer, and it has been originally established in 1973 at the Michigan Cancer Foundation from a pleural effusion taken from a woman with metastatic breast cancer. Butanolic extract of the roots of P. paniculata showed cytotoxic effect MCF-7 cell line, as determined with crystal violet assay, cellular death with acridine orange/ethidium bromide staining, and cell proliferation with immunocytochemistry of bromodeoxyuridine (BrdU). Subcellular alterations were evaluated by electron microscopy. Cells treated with butanolic extract showed degeneration of cytoplasmic components and profound morphological and nuclear alterations. The results show that this butanolic extract indeed presents cytotoxic substances, and its fractions merit further investigations.

Phytochemical study of Solanum lycocarpum (St. Hil) unripe fruit and its effects on rat gestation.

Solanum lycocarpum, St. Hil (Solanaceae) is a common native shrub in the Brazilian cerrado. The fruits are used in folk medicine as a hypoglycaemic agent in the management of diabetes, obesity and to decrease cholesterol levels. In this study the glycoalkaloids, solamargine and solasonine, were isolated from unripe fruits of S. lycocarpum. To evaluate the effects of the fruits on gestation, pregnant rats (n=25) were fed from day 6 to 22 with chow containing 10% of dried and ground unripe fruits. The control group (n=21) received regular chow. During and after the treatment period the dams showed reduced body weight and slower body weight gain, even with no change in food and water intake, evidencing mild maternal toxicity. Gestation was not significantly impaired, although experimental fetuses presented reduced body length at birth. Also, 20% of the treated dams showed one or two dead pups at birth. On day 22 of gestation and on post-natal day 1, the levels of metabolites of the sex hormones oestradiol, progesterone and testosterone were measured in faeces by radioimmunoassay. On post-natal day 1, tissue portions from the dams were collected for histopathological evaluation. No alterations were detected in either study. The results suggest that S. lycocarpum fruit did not impair gestation, however, it did promote mild maternal toxicity and mild fetotoxic effects if ingested as a food source during the gestation period. This study has implications for pregnant women, who employ phytotherapeutic formulations under the impression that they are harmless.

Pfaffia paniculata (Brazilian ginseng) methanolic extract reduces angiogenesis in mice.

Pfaffia paniculata (Brazilian ginseng) roots have been indicated for the treatment of several diseases. Our studies have shown that P. paniculata roots present antineoplastic effects and cancer chemopreventive activity in a mouse hepatocarcinogenesis model. The purpose of this study was to investigate the effects of the Brazilian ginseng on corneal angiogenesis in mice. We first conducted a toxicological study employing 250, 500, or 1000mg/kg/day of the methanolic extract of P. paniculata roots by gavage to BALB/c mice. Animals did not lose weight during the treatment nor presented histopathological alterations. The effect of this root on angiogenesis in the cornea of BALB/c mice was then assessed. Male mice were treated, by gavage, once a day, with doses of 250, 500, or 1000mg/kg of methanolic extract of P. paniculata powdered root for 10 days; filtered water was used as control. Corneal cauterization was accomplished by the contact of a silver nitrate crystal on the central area of the cornea, in the 5th day of treatment with P. paniculata, which continued thereafter; the animals were euthanized on the 6th day after cauterization. Newly formed blood vessels were filled with India ink, and the corneas were routinely processed. Blood vessels were quantified in an image analysis system. A smaller total area of neovascularization in the mouse cornea was observed in animals treated with 1000mg/kg of the methanolic extract of P. paniculata. These results indicate an antiangiogenic effect of this extract. The mechanisms of this antiangiogenic activity of P. paniculata should be further investigated.

Glaziovianin A, a new isoflavone, from the leaves of Ateleia glazioviana and its cytotoxic activity against human cancer cells.

A new isoflavone, named glaziovianin A (1), was isolated from the leaves of Ateleia glazioviana (Legminosae) by means of a cytotoxicity-guided fractionation procedure against HL-60 leukemia cells. The chemical structure of 1 was determined by analysis of its extensive spectroscopic data. Glaziovianin A (1) displayed differential cytotoxicities in the Japanese Foundation for Cancer Research 39 cell line panel assay. The pattern of the differential cytotoxicities of 1 was found to correlate to that shown by TZT-1027, suggesting that 1 inhibited tubulin polymerization as an action mechanism. Although 1 had little influence on microtubule networks in interphase cells, 1-treated cells showed abnormal structures with unaligned chromosomes.

Assessment of the perinatal effects of maternal ingestion of Ipomoea carnea in rats.

It is believed that Ipomoea carnea toxicosis induces abnormal embryogenesis in livestock. Studies on rats treated with I. carnea aqueous fraction (AF) during gestation, revealed litters with decreased body weight, but the characteristic vacuolar lesions promoted by swainsonine, its main toxic principle, were observed only in young rats on postnatal day (PND) 7. However, these alterations could have resulted as consequence of swainsonine placental passage and/or damage or even ingestion of the contaminated milk by pups. Thus, this perinatal work was performed to verify the transplacental passage of swainsonine and its excretion into milk employing the cross-fostering (CF) procedure as a tool of study. Females were treated with AF or vehicle during gestation and after birth pups were fostered between treated and untreated dams. Pup body weight gain (BWG) and histopathology to observe vacuolar degeneration were performed on PND 3 and 7. In addition, swainsonine detection was performed in amniotic fluid and milk from rats treated with the AF during gestation or lactation. BWG was significantly lower only in pups from mothers treated with the plant and fostered to other treated mothers (AF-AF group of pups). The histopathology revealed that pups from treated mothers fostered to untreated ones showed the characteristic vacuolar lesions; however, the lesions from the AF-AF pups were more severe in both periods evaluated. Amniotic fluid and milk analysis revealed the presence of swainsonine excretion into these fluid compartments. Thus, the results from CF and the chemical analysis allowed concluding that swainsonine passes the placental barrier and affects fetal development and milk excretion participates in I. carnea perinatal toxicosis.

Crystal-associated cholangiopathy in sheep grazing Brachiaria decumbens containing the saponin protodioscin

An outbreak of hepatogenous photosensitization is reported in a flock of 28 sheep grazing Brachiaria decumbens in Mato Grosso do Sul State, Central-Western Brazil. Seven lambs and an adult sheep were affected and 6 of them died. Two surviving affected lambs and one lamb without clinical signs had increased serum values of gamma glutamyltransferase, bilirubin, and cholesterol. In two adult unaffected sheep those parameters were within normal values. An adult sheep submitted to necropsy presented moderate body condition, unilateral corneal opacity, drying of the muzzle, moderate jaundice, increased lobular pattern of the liver, and a distended gallbladder. Histological lesions were epithelial degeneration, necrosis, and hyperplasia of small bile ducts. Mild amounts of foamy macrophages were observed, mainly in the centroacinar zone. Diffuse swelling and vacuolation were observed in hepatocytes. Crystal negative images were found within bile ducts, foamy macrophages, and the lumen of some renal tubules. The heart showed multifocal areas of degeneration and necrosis of the muscle fibers. Pasture samples (Brachiaria decumbens) contained 2.36 percent of protodioscin. No Pithomyces chartarum spores were found in the pasture. Samples from a similar neighboring B. decumbens pasture grazed by cattle without photosensitization contained 1.63 percent of protodioscin isomers. Outbreaks of photosensitization caused by Brachiaria spp. are common in cattle in the Brazilian Cerrado (savanna) with about 51 million hectares of Brachiaria spp pastures. Sheep farming has been recently developed in this region, and the number of sheep is increasing significantly. Because sheep are more susceptible than cattle to lithogenic saponins, poisoning by Brachiaria should be an important limiting factor for the sheep industry.

Um surto de fotossensibilização hepatógena é descrito em um rebanho de 28 ovinos mantidos em pastagem de Brachiaria decumbens no Estado de Mato Grosso do Sul, região Centro-Oeste do Brasil. Sete cordeiros e uma ovelha adulta foram afetados e seis deles morreram. Dois cordeiros com sinais clínicos e um cordeiro, aparentemente sem sinais clínicos, apresentaram elevação na atividade sérica de gama glutamil-transferase, e nos valores de bilirrubina e colesterol. Em dois animais adultos que não apresentavam sinais clínicos, esses parâmetros estavam dentro dos valores normais. Uma ovelha adulta submetida à necropsia apresentou moderada condição corporal, opacidade de córnea unilateral, focinho ressecado, moderada icterícia, padrão lobular hepático evidente e vesícula biliar acentuadamente distendida e repleta. As lesões histológicas consistiram de degeneração epitelial, necrose e hiperplasia de ductos biliares. Havia discreta quantidade de macrófagos espumosos, principalmente na região centroacinar, e tumefação e vacuolização difusas dos hepatócitos. Foram observadas imagens negativas de cristais dentro dos ductos biliares, macrófagos espumosos e lúmen de alguns túbulos renais. O coração apresentou áreas multifocais de degeneração e necrose de fibras musculares. Amostras da pastagem (B. decumbens) apresentaram 2,36 por cento de protodioscina e nenhum esporo de Pithomyces chartarum foi encontrado na pastagem. Amostras de uma pastagem vizinha semelhante (B. decumbens), que havia sido pastejada por bovinos que não desenvolveram fotossensibilização, apresentaram 1,63 por cento de protodioscina. Surtos de fotossensibilização causados por espécies de Brachiaria são comuns em bovinos no Cerrado brasileiro onde há aproximadamente 51 milhões de hectares plantados com essa gramínea. A criação de ovinos nesta região ganhou recente impulso e o número de ovinos vem aumentando significativamente. Como os ovinos são mais suscetíveis que os bovinos à intoxicação...