Thesis - Wisconsin. / Title from PDF title page (viewed Nov. 10, 2008). Includes bibliographical references (p. 190-196). Online version of the print original.

Thesis--Wisconsin. / Includes bibliographical references (leaves 190-196).

Made available in DSpace on 2014-06-12T15:53:46Z (GMT). No. of bitstreams: 2 arquivo4867_1.pdf: 2673938 bytes, checksum: 936cca7358e1df135e6731cdb5f8bd3c (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2006 / As lectinas são proteínas ubíquas na natureza que se ligam reversívelmente a mono, oligo, polissacarídeos e glicoconjugados. Não apresentam atividade catalítica e ao contrário dos anticorpos, não são produtos de uma resposta imunológica. O objetivo do presente trabalho foi avaliar a extração e a reextração de uma lectina purificada por cromatografia de troca iônica (CrataBL) e do extrato bruto (EB) da entrecasca de Crataeva tapia utilizando o sistema de micelas invertidas, constituídas pelo surfactante dioctilsulfosuccinato de sódio (AOT) em isooctano. A entrecasca de Crataeva tapia foi coletada na região da cidade do Recife (Pernambuco, Brasil) e o extrato [10 % (p/v) em 150 mM NaCl] foi obtido por trituração e agitação durante 16 h a 4 oC, filtrado em gaze e centrifugado (4.000 x g durante 15 min). O sobrenadante obtido foi denominado de extrato bruto (EB). Os fatores que afetam a extração da proteína tais como: tempo de contato de agitação (5 - 20 min), força iônica, incluindo o tipo de sal (NaCl, KCl e CaCl2) e concentração (30 300 mM), pH da fase aquosa (pH 3,0 12,0) e concentração do surfactante (5 - 100 mM AOT), foram investigados. Os parâmetros avaliados para a reextração foram: pH da fase aquosa (pH 5,0 7,0) e força iônica (50 - 1000 mM de KCl) tendo sido adicionado ao sistema 5% Butanol. Os parâmetros velocidade de agitação (900 rpm), temperatura (25oC) e concentração de proteína (0,374 mg/ml), foram mantidos constantes em todos os experimentos. Os melhores resultados para extração foram obtidos com 5 min de tempo de contanto entre as duas fases, 30 mM de NaCl, tampão citrato/fostato pH 5,5 e 5 mM de AOT, onde foi possível obter extrações protéicas de 100 % e 70 % para CrataBL e EB, respectivamente. Para a reextração, as melhores condições foram, tampão citrato/fosfato 10 mM, pH 5,5 acrescido de 1000 mM de KCl, onde foi possível obter uma recuperação protéica de 45,25 % (CrataBL) e 80,65 % (EB) com 50 % da atividade hemaglutinante para ambas as amostras. As amostras obtidas com as melhores condições de extração e reextração aplicadas ao EB revelaram apenas uma banda na PAGE para proteínas básicas e duas bandas no SDSPAGE. A cromatografia de gel filtração (AKTA) indicou dois picos: um de 40 KDa e outro de 29 KDa. A natureza oligomérica da lectina foi detectada por cromatografia de filtração em gel e SDS-PAGE. A comparação dos perfis cromatográficos de filtração em gel do EB com o da lectina purificada, indicaram a eficiência do sistema de micelas invertidas na purificação da lectina

Dormancy characteristics and optimum conditions for germination of Cleome gynandra seeds have not been explained. Seed storage proteins were extracted, analysed with SDS-PAGE and sequenced. Seed proteins of Cleome were characterised by comparison with those of wild mustard (Brassica kaber). Wild mustard showed seed proteins composed of two α-chains of molecular weight (24-32 kDa) and another two β-chains of 18-22 kDa. The seed proteins of Cleome comprised two α-chain polypeptides of molecular weight (25-30 kDa), two β-chain polypeptides of molecular weight (18-20 kDa) and a smaller β-chain of 13-15 kDa. The storage proteins occurred in the seeds as dimeric complexes of molecular weight 40-65 kDa, which were broken into polypeptide chains of approximately 20 and 30 kDa by the reducing . action of DTT. Comparison with proteins in the proteome library and similarity index further confirmed that the seed proteins of Cleome had similarities with those of wild mustard. Two dimensional SDS-PAGE showed that the two species have nine similar polypeptides and four different ones. Events associated with dormancy release during seed germination still require explanation. Seeds of Cleome are characterised by low germination and there has been no explanation for this. Changes in protein expression during germination of Cleome in the presence or absence of light and at constant or alternating temperatures were examined. The germination of Cleome seeds at 20 degrees C was inhibited by light, but it was improved at 20 degrees C in darkness. There was no photoinhibition when seeds were germinated at constant 30 degrees C or alternating 20/30 degrees C (16 h night and 8 h day) for 10 days. Four proteins were observed to decrease in expression as germination progressed, but remained unchanged during photoinhibition. Photoinhibition was expressed more in seeds that were harvested late, after the pods had turned brown. These seeds showed a fifth, low molecular weight protein (13 kDa) that was absent from the immature seeds and embryos. Photinhibition is a pseudo-dormancy condition during which seed storage proteins are not utilised and the seed coat could partially play a role in it. The temperatures for the germination of Cleome in darkness have been determined. However, prior to this study the effects of temperature, light and pre-germination treatments (chilling, scarification, hydration and germination in the presence of KN0(3) or GA(3) on the germination of the seeds of this species have not been investigated. Seeds were germinated for 10 days and the final count of germination was used to determine seed performance. The highest germination percentage (60% and 80%, for a 2-year old and a l-year old seed lot, respectively) of untreated seeds was achieved when alternating temperatures of 20/30 degrees C (16 h/S h) in the dark or constant 30 degrees C in the dark were used. Among the pre-germination treatments, only scarification (puncturing of seeds at the radicle end) improved germination. Seeds were found to be negatively photoblastic, and the phenomenon was more pronounced when they were germinated at 20 degrees C and 12 h photoperiod or longer. Germination of photoinhibited seeds was, however, improved by treatment with GA(3) It is recommended that the germination of Cleome be undertaken under conditions of darkness and at either alternating 20/30 degrees C or continuous 30 degrees C. Seed lot vigour and seedling vigour are two important seed quality aspects that are used in defining the seed germination process. Seed germination is appropriately characterised by radicle protrusion and the attainment of normal seedling structures. However, the international rules for testing seeds combine radicle protrusion and normal seedling attainment in separating seed germination into the first and final counts. The challenge to a seed analyst testing the germination of a species whose first and final counts are unknown is that there is no statistical guideline to determine these important stages of seed germination. Cauliflower and broccoli, for which the first and final counts are published in the international rules for testing seeds and Cleome, for which there is no data on the first and final counts, were examined to determine the statistical significances of the first and final counts. Analysis of variance, logistic regression, 'broken-stick' regression models and survival analysis procedures were used. Analysis of variance showed that there were no differences between the germination percentages on the fourth, fifth and seventh days of germination. Low and stable standard deviations were recorded when evaluating germination after the fourth day. The germination curves of broccoli and cauliflower did not fit the Gompertz curve but fitted the exponential curve. The broken-stick model 'broke' the cumulative germination curve for the Cleome seed lots into two linear curves that were significantly different, but failed to break those for broccoli and cauliflower. However, this study confirmed the first and final counts for broccoli and cauliflower as determined by the international rules for testing seeds. Broken-stick modelling and life table analyses confirmed the fourth day as being appropriate to determine the first count for Cleome germination. There was no evidence of further seed germination after the seventh day as shown by probability density and hazard rate. It is suggested that for Cleome, the 'first count' and 'final count' be performed on the fourth and seventh day of the germination, respectively. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

INTRODUÇÃO: Os corticosteróides são considerados padrão-ouro no tratamento da asma, porém, existem asmáticos graves que não obtém controle dos sintomas, o que suscita a busca por novas terapias. Os inibidores de proteinases têm sido estudados no controle de diversos processos inflamatórios, dentre estes, encontra-se Crataeva tapia Bark Lectin (CrataBL). OBJETIVO: Avaliar se a proteina bifuncional de planta, CrataBL, que tem função lectínica e se liga a carboidratos, modula a hiperresponsividade brônquica à metacolina, inflamação, remodelamento e estresse oxidativo nas vias aéreas e nos septos alveolares de camundongos com inflamação pulmonar alérgica crônica. MÉTODOS: Trinta e dois camundongos machos Balb-c SPF (6-7 semanas, 25-30 g) foram divididos em 4 grupos: C (controle), OVA (sensibilizados - 50 ug de ovalbumina intraperitoneal (i.p) nos dias 0 e 14 e desafiados - 1% de ovalbumina nos dias 22, 24, 26, 28); C+CR (controle tratados com CrataBL - 2 mg/kg/i.p dos dias 22 a 28); OVA+CR (sensibilizados e desafiados com ovalbumina e tratados com CrataBL - 2 mg /kg/i.p dos dias 22 a 28). No dia 29, realizamos: (i) hiperresponsividade à metacolina - resposta máxima de resistência (Rrs) e elastância (Ers) do sistema respiratório; (ii) quantificação do número total de células, macrófagos, linfócitos e células polimorfonucleares no fluido do lavado broncoalveolar (FLBA); (iii) análise histopatológica do pulmão por morfometria para quantificação de eosinófilos, fração de volume de fibras colágenas e elásticas; (iiii) imunohistoquímica para quantificação de células positivas para IL-4, IL-5, IL-13, IFN-y, MMP-9, TIMP-1, TGF-beta, iNOS, NF-kB e fração de volume de 8-iso-PGF2alfa nas vias aéreas e nos septos alveolares e ELISA para quantificação da concentração de IL-4, IL-5 e IFN-y. A significância foi considerada quando p < 0,05. RESULTADOS: Houve atenuação da resposta máxima de Rrs e Ers no grupo OVA+CR comparado ao grupo OVA (p < 0,05). O tratamento com CrataBL nos animais sensibilizados atenuou o número de células totais, macrófagos, linfócitos e células polimorfonucleares no FLBA, o número de eosinófilos, células positivas para IL-4, IL-5, IL-13, IFN-y, iNOS, MMP-9, TIMP-1, TGF-beta, NF-kB e fração de volume de 8-iso-PGF2alfa, fibras colágenas e elásticas tanto nas vias aéreas quanto nos septos alveolares quando comparados ao grupo OVA. O tratamento com CrataBL atenuou os níveis de concentração de IL-4, IL-5 e IFN-y em comparação ao grupo OVA (p < 0,05), a partir do método ELISA. CONCLUSÕES: CrataBL atenuou a hiperresponsividade brônquica, a inflamação, o remodelamento e o estresse oxidativo nesse modelo experimental de inflamação pulmonar alérgica crônica. Contudo, mais estudos são necessários para verificar se este inibidor pode ser uma potencial ferramenta terapêutica para a asma / RATIONALE: Corticosteroids are considered the gold standard in the treatment of asthma, however, there are severe asthmatics who do not get control of symptoms, which raises the search for new therapies. The proteinase inhibitors have been studied in the control of various inflammatory processes, including such inhibitors is Crataeva tapia Bark Lectin (CrataBL). OBJECTIVE: To evaluate the proteinase inhibitor CrataBL modulates the bronchial responsiveness to methacholine, inflammation, remodeling and activation of oxidative stress in the airways and alveolar septa of mice with chronic allergic pulmonary inflammation. METHODS: Thirty two SPF Balb-c male mice (6-7 weeks, 25-30 g) were divided into 4 groups: control (C), OVA (sensitized - 50 ug ovalbumin intraperitoneal (i.p) on days 0 and 14 and challenged - 1% ovalbumin on days 22, 24, 26, 28); C+CR (control treated with CrataBL - 2 mg / kg / i.p of 22 to 28); OVA+CR (sensitized and challenged with ovalbumin and treated with CrataBL - 2 mg / kg / i.p from day 22 to 28). On the 29th, we held: (i) hyperresponsiveness to methacholine and maximal responses were obtained resistance (Rrs) and elastance (Ers) of the respiratory system; (ii) quantification of total cells, macrophages, polymorphonuclear cells and lymphocytes in bronchoalveolar lavage fluid (BALF); (iii) histopathological analysis of the lungs by morphometry to quantify the eosinophils, volume fraction of collagen and elastic fibers; (iiii) immunohistochemistry for quantification of positive IL-4, IL-5, IL-13, IFN-y, MMP-9, TIMP-1, TGF-beta, iNOS, NFk-beta cells and volume fraction of 8-iso-PGF2? in airway and alveolar septa and ELISA for quantification of concentration of IL-4, IL-5 e IFN-y (p < 0.05). RESULTS: There was attenuation of the maximal response Rrs and Ers in the OVA+CR group compared to OVA (p < 0.05). Treatment with CrataBL in sensitized animals attenuated the number of total cells, macrophages, lymphocytes, and polymorphonuclear cells in BALF, the number of eosinophil positive IL-4, IL-5, IL-13, IFN-y, iNOS, MMP -9, TIMP-1, TGF-beta, NF-kB cells and volume fraction of 8-iso-PGF2alfa, collagen and elastic fibers in both the airways and alveolar septa compared to OVA group. Treatment with CrataBL attenuated concentration levels of IL-4, IL-5 and IFN-y compared to OVA group (p < 0.05) from the ELISA method.CONCLUSIONS: CrataBL attenuated bronchial hyperresponsiveness, inflammation, remodeling and oxidative stress in this experimental model of chronic allergic pulmonary inflammation. However, more studies are needed to determine if this inhibitor can be a potential therapeutic tool for asthma

INTRODUÇÃO: Os corticosteróides são considerados padrão-ouro no tratamento da asma, porém, existem asmáticos graves que não obtém controle dos sintomas, o que suscita a busca por novas terapias. Os inibidores de proteinases têm sido estudados no controle de diversos processos inflamatórios, dentre estes, encontra-se Crataeva tapia Bark Lectin (CrataBL). OBJETIVO: Avaliar se a proteina bifuncional de planta, CrataBL, que tem função lectínica e se liga a carboidratos, modula a hiperresponsividade brônquica à metacolina, inflamação, remodelamento e estresse oxidativo nas vias aéreas e nos septos alveolares de camundongos com inflamação pulmonar alérgica crônica. MÉTODOS: Trinta e dois camundongos machos Balb-c SPF (6-7 semanas, 25-30 g) foram divididos em 4 grupos: C (controle), OVA (sensibilizados - 50 ug de ovalbumina intraperitoneal (i.p) nos dias 0 e 14 e desafiados - 1% de ovalbumina nos dias 22, 24, 26, 28); C+CR (controle tratados com CrataBL - 2 mg/kg/i.p dos dias 22 a 28); OVA+CR (sensibilizados e desafiados com ovalbumina e tratados com CrataBL - 2 mg /kg/i.p dos dias 22 a 28). No dia 29, realizamos: (i) hiperresponsividade à metacolina - resposta máxima de resistência (Rrs) e elastância (Ers) do sistema respiratório; (ii) quantificação do número total de células, macrófagos, linfócitos e células polimorfonucleares no fluido do lavado broncoalveolar (FLBA); (iii) análise histopatológica do pulmão por morfometria para quantificação de eosinófilos, fração de volume de fibras colágenas e elásticas; (iiii) imunohistoquímica para quantificação de células positivas para IL-4, IL-5, IL-13, IFN-y, MMP-9, TIMP-1, TGF-beta, iNOS, NF-kB e fração de volume de 8-iso-PGF2alfa nas vias aéreas e nos septos alveolares e ELISA para quantificação da concentração de IL-4, IL-5 e IFN-y. A significância foi considerada quando p < 0,05. RESULTADOS: Houve atenuação da resposta máxima de Rrs e Ers no grupo OVA+CR comparado ao grupo OVA (p < 0,05). O tratamento com CrataBL nos animais sensibilizados atenuou o número de células totais, macrófagos, linfócitos e células polimorfonucleares no FLBA, o número de eosinófilos, células positivas para IL-4, IL-5, IL-13, IFN-y, iNOS, MMP-9, TIMP-1, TGF-beta, NF-kB e fração de volume de 8-iso-PGF2alfa, fibras colágenas e elásticas tanto nas vias aéreas quanto nos septos alveolares quando comparados ao grupo OVA. O tratamento com CrataBL atenuou os níveis de concentração de IL-4, IL-5 e IFN-y em comparação ao grupo OVA (p < 0,05), a partir do método ELISA. CONCLUSÕES: CrataBL atenuou a hiperresponsividade brônquica, a inflamação, o remodelamento e o estresse oxidativo nesse modelo experimental de inflamação pulmonar alérgica crônica. Contudo, mais estudos são necessários para verificar se este inibidor pode ser uma potencial ferramenta terapêutica para a asma / RATIONALE: Corticosteroids are considered the gold standard in the treatment of asthma, however, there are severe asthmatics who do not get control of symptoms, which raises the search for new therapies. The proteinase inhibitors have been studied in the control of various inflammatory processes, including such inhibitors is Crataeva tapia Bark Lectin (CrataBL). OBJECTIVE: To evaluate the proteinase inhibitor CrataBL modulates the bronchial responsiveness to methacholine, inflammation, remodeling and activation of oxidative stress in the airways and alveolar septa of mice with chronic allergic pulmonary inflammation. METHODS: Thirty two SPF Balb-c male mice (6-7 weeks, 25-30 g) were divided into 4 groups: control (C), OVA (sensitized - 50 ug ovalbumin intraperitoneal (i.p) on days 0 and 14 and challenged - 1% ovalbumin on days 22, 24, 26, 28); C+CR (control treated with CrataBL - 2 mg / kg / i.p of 22 to 28); OVA+CR (sensitized and challenged with ovalbumin and treated with CrataBL - 2 mg / kg / i.p from day 22 to 28). On the 29th, we held: (i) hyperresponsiveness to methacholine and maximal responses were obtained resistance (Rrs) and elastance (Ers) of the respiratory system; (ii) quantification of total cells, macrophages, polymorphonuclear cells and lymphocytes in bronchoalveolar lavage fluid (BALF); (iii) histopathological analysis of the lungs by morphometry to quantify the eosinophils, volume fraction of collagen and elastic fibers; (iiii) immunohistochemistry for quantification of positive IL-4, IL-5, IL-13, IFN-y, MMP-9, TIMP-1, TGF-beta, iNOS, NFk-beta cells and volume fraction of 8-iso-PGF2? in airway and alveolar septa and ELISA for quantification of concentration of IL-4, IL-5 e IFN-y (p < 0.05). RESULTS: There was attenuation of the maximal response Rrs and Ers in the OVA+CR group compared to OVA (p < 0.05). Treatment with CrataBL in sensitized animals attenuated the number of total cells, macrophages, lymphocytes, and polymorphonuclear cells in BALF, the number of eosinophil positive IL-4, IL-5, IL-13, IFN-y, iNOS, MMP -9, TIMP-1, TGF-beta, NF-kB cells and volume fraction of 8-iso-PGF2alfa, collagen and elastic fibers in both the airways and alveolar septa compared to OVA group. Treatment with CrataBL attenuated concentration levels of IL-4, IL-5 and IFN-y compared to OVA group (p < 0.05) from the ELISA method.CONCLUSIONS: CrataBL attenuated bronchial hyperresponsiveness, inflammation, remodeling and oxidative stress in this experimental model of chronic allergic pulmonary inflammation. However, more studies are needed to determine if this inhibitor can be a potential therapeutic tool for asthma

Thesis (M. Sc.(Agronomy)) -- University of Limpopo, 2017. / Cleome gynandra is increasingly becoming an important strategy for achieving food and nutrition security among rural households in many developing countries. Root-knot (Meloidogyne species) nematodes, with limited nematode management strategies, limit the successful production of this vegetable crop. Nemafric-BL and Nemarioc-AL phytonematicides are separately being developed in South Africa for sustainable crop production systems. However, the two products have not been simultaneously tested for managing the notorious Meloidogyne species and absorption of phosphorus, with a combination of Vesicular arbuscular mycorrhiza (VAM). The objective of this study therefore was to determine the interactive effects of VAM and each of the two phytonematicides on nutrient content, growth of C. gynandra. A 2 × 2 × 2 factorial experiment, with the first, second and third factors being VAM (V), nematode (N) and Nemafric-BL phytonematicide (P). The eight treatments included (1) untreated control (V0N0P0), (2) nematodes alone (V0N1P0), (3) VAM alone (V1N0P0) (4) Nemarioc-AL phytonematicide alone (V0N0P1), (5) V1N1P0, (6) V0N1P1, (7) V1N0P1 and (8) V1N1P1, were laid out in a randomised complete block design, with ten replications. The same layout experiment was done for the Nemarioc-AL phytonematicide trial which had a similar layout. Seedlings were irrigated with 250 ml chloride-free tapwater every other day for 56 days. Multifeed and NPK (2:3:2(22) fertilisers were applied at transplanting. The second order interaction (V1N1P1), was highly significant (P ≤ 0.01) for plant height contributing 54% in TTV (Total Treatment Variation) of the variable. Among the main factors (N, P and V), only nematode had highly significant effects on stem diameter. All interactions of VAM, nematode and Nemarioc-AL phytonematicide and main factors each had no significant effect on Cleome. The second order (V1N1P1) and the first order interaction (V1N1P1) did not have significant effects on the three nutrient elements except for the first order interaction (V1N0P1) which was significant on foliar Zn contributing 42% in TTV of the variable. Also nematode had highly significant effect on foliar K and significant effect on foliar Zn contributing 49 and 31% in TTV of the respective variables. Using the two-way table, VAM and Nemafric-BL phytonematicide each increased foliar Zn by 27% and 29%, respectively. The second and first order interactions of VAM, N and Nemarioc-AL phytonematicide and the main factors did not have significant effect on foliar K, Fe and Zn. The second order interaction of VAM, nematode and Nemafric-BL phytonematicide had significant effects on gall rating, contributing 2% in TTV of the variable. VAM, nematode and Nemarioc-AL phytonematicide showed that the second and first order interaction except for V1N0P1 interaction on gall rating, were not significant for nematode variables. The V1N0P1 interaction contributed 20% in TTV of gall rating. Using a two-way table, VAM and phytonematicide each increased root galls by 7% and 74%, respectively. Combined, VAM and phytonematicide reduced root galls by 64%. The innovative products interacted together and that Nemafric-BL and Nemarioc-AL phytonematicides and VAM alone could be used in managing nematodes. / National Research Foundation, Agricultural Research Council-Universities Collaboration Centre

O objetivo deste trabalho foi avaliar se a proteína bifuncional de planta, CrataBL, que tem lectina e as propriedades inibidoras de enzima, modula alterações de mecânica pulmonar, inflamatórias e remodelamento induzidas por elastase intratraqueal em camundongos. Métodos: 36 camundongos C57BL6 receberam elastase (0,025 mg) por instilação intratraqueal (grupo ELA e ELA-CrataBL). Os grupos controles receberam salina (grupo SAL e SAL-CrataBL). Os camundongos foram tratados com instilação intraperitoneal de CrataBL (2mg/kg) nos dias 1, 14 e 21 após a instilação intratraqueal de elastase (grupo SAL-CrataBL e ELA-CrataBL) os animais controle receberam salina no mesmo volume. No dia 28, os camundongos foram anestesiados, ventilados mecanicamente e foram analisados a resistência e elastância do sistema respiratório (Ers e Rrs), elastância e resistência tecidual (Htis e Gtis), resistência das vias aéreas (Raw) e óxido nítrico exalado (NOex). Após, o lavado broncoalveolar (LBA) foi realizado, os pulmões foram retirados e por morfometria, e foram quantificados o intercepto linear médio (Lm), a quantidade de neutrófilos, células positivas para TNF-alfa, fibras colágenas, elásticas, células positivas para MMP-9, MMP-12, TIMP-1, eNOS e iNOS e isoprostano no parênquima pulmonar e vias aéreas. No parênquima foram avaliados os macrófagos nos septos alveolares e nas vias aéreas, foram também avaliadas as células para MUC-5. Resultados: No grupo ELA houve um aumento na Ers, Raw, Gtis, Htis, Lm, NOex, nas células totais, macrófagos, neutrófilos, eosinófilos e linfócitos no LBA em relação aos controles (p < 0,05), sendo que Raw, diminuiu também nos grupos SAL-CrataBL e ELA-CrataBL. Nos grupos tratados com CrataBL houve uma diminuição de Ers (37,0±2,2 cmH2O/L), Htis (37,9±3,5 cmH2O/ml/s), ENO (14,7±0,7 ppb), comparativamente ao grupo ELA (p < 0,05). No LBA houve atenuação de neutrófilos (0,003±0,001 104células/ml), linfócitos (0,003±0,001 104células/ml) e de Lm (54,6±6,0 mm). Complementando a avaliação, no grupo que recebeu elastase houve um aumento no número de macrófagos (22,88 +- 2,24 células/104um2), neutrófilos (1,18 +- 0,15 células/10 4um2), células positivas para TNF-ala (12,52 +- 0,42 células/104um2) no parênquima pulmonar. Nas alterações de remodelamento no parênquima pulmonar, houve um aumento da proporção de volume de fibras colágenas (11,5 +- 0,11%), elásticas (0,5 +- 0,03%), na quantidade de células positivas para MMP-9 (18,59 +- 1,87 células/104?m2), MMP-12 (20,17 +- 1,92 células/104?m2), TIMP-1 (14,42 +- 2,05 células/104um2) em comparação com os controlos (p < 0,001). No estresse oxidativo, houve um aumento de eNOS (13,15 +- 0,40 células/104um2), iNOS (10,49 +- 0,65 células/104um2) e isoprostano (18,11 =- 5,38%). O tratamento CrataBL (grupo ELA-CrataBL) reduziu no parênquima pulmonar a quantidade de macrófagos (9,58 +- 1,36 células/104um2), neutrófilos (0,75 +- 0,1 células/104um2), células positivas para TNF-alfa (10.4±0,49 células/104?m2), fibras colágenas (10,8 +- 0,13%), elásticas (0,3 +- 0,02%), a quantidade de células positivas para a MMP-9 (10,35±0,65 células/104um2), MMP-12 (14,15±0,59 células/104um2), TIMP-1 (9,89 +- 2,79 células/104um2), MUC-5 (3,56 +- 0,54 células/104um2), eNOS (6.98 +- 0.32 células/104um2) e iNOS (6,21 +- 0,42 células/104um2) e isoprostano (8,96 +- 3,08 %) em relação ao grupo ELA (p < 0,001). Nas vias aéreas também ocorreu um aumento significativo de neutrófilos (5,97 +- 1,03 células/104um2), células positivas para TNF-alfa (15,82 +- 1,03 células/104um2). Nas alterações de remodelamento pulmonar nas vias aéreas também ocorreu um aumento da proporção de volume de fibras colágenas (8,73 +- 2,59%), elásticas (2,56 +- 0,18%), na quantidade de células positivas para MMP-9 (14,86 +- 1,77 células/104um2), MMP-12 (18,56 +- 1,79 células/104um2), TIMP-1 (1,31 +- 0,12 células/104um2) e MUC-5 (7,09 +- 1,71 células/104um2) em comparação com os controlos (p < 0,001). No estresse oxidativo, houve um aumento de células positivas para eNOS (3,09 +- 0,08 células/104um2), iNOS (5,4 +- 0,3 células/104um2) e isoprostano (18,11 +- 5,38%) em comparação com os controlos (p < 0,001). O tratamento CrataBL (grupo ELA-CrataBL) reduziu nas vias aéreas a quantidade de neutrófilos (4,62 +- 0,61 células/104um2), TNF- alfa (14,30 +- 1,28 células/104um2), fibras colágenas (7,80 +- 1,37%), elásticas (1,4 +- 0,13%), a quantidade de células positivas para a MMP-9 (9,93 +- 1,39 células/104um2), MMP-12 (12,06 +- 1,15 células/104um2), TIMP-1 (0,73 +- 0,05 células/104?m2), MUC-5 (3,56 +- 0,54 células/104um2), eNOS (1,89 +- 0,16 células/104um2) e iNOS (4,3 +- 0,31 células/104um2), isoprostano (7,34 +- 2,31%) em relação ao grupo ELA (p < 0,001). Conclusão: CrataBL atenua as alterações de mecânica pulmonar, lavado bronco alveolar, responsividade inflamatória, controle do remodelamento e estresse oxidativo induzidas pela elastase. Embora mais estudos devam ser realizados, esta proteína bifuncional pode contribuir como potencial ferramenta terapêutica para o tratamento da DPOC / The aim of this study was to evaluate whether the bifunctional protein plant, CrataBL, which has lectin and enzyme inhibitory properties, modulates changes in lung mechanics, inflammatory and remodeling induced by intratracheal elastase in mice.Methods : 36 C57/Bl6 mice received elastase (0.025 mg) by intratracheal (group ELA and ELA-CrataBL). Control groups received saline (group SAL and SAL-CrataBL).The mice were treated with intraperitoneal instillation of CrataBL (2mg/kg) on days 1, 14 and 21 after intratracheal instillation of elastase (group SAL-CrataBL and ELA-CrataBL), control animals received saline in the same volume. On day 28, the mice were anesthetized and mechanically ventilated were analyzed resistance and respiratory system elastance (Ers and Rrs), elastance and tissue resistance (Htis and Gtis), airway resistance (Raw) and exhaled nitric oxide (ENO). After the bronchoalveolar lavage (BAL) was performed, the lungs were removed and morphometry were quantified and the linear intercept mean (Lm), the number of neutrophils, positive cells for TNF-alfa, collagen fibers, positive cells for MMP-9, MMP-12, TIMP-1, eNOS, iNOS and isoprostane in lung parenchyma and airways. Parenchyma was also evaluated macrophages in the alveolar septa. Airway was also evaluated MUC-5 cells. Results: In group ELA was an increase in Ers, Raw, Gtis, Htis, Lm, ENO, in total cells, macrophages, neutrophils, eosinophils and lymphocytes in BAL compared to controls (p < 0.05), and Raw, decreased in both groups SAL-CrataBL and ELA-CrataBL. In the groups treated with CrataBL there was a decrease in Ers (37.0±2.2 cmH2O/L) Htis (37 9±3.5 cmH2O/ml/s) and ENO (14.7±0.7 ppb) compared to the ELA group (p < 0.05). In BAL there was attenuation of neutrophils (0.003±0.001 104cells/ml), lymphocytes (0.003±0.001 104cells/ml) and Lm (54.6±6.0 mm). Complementing the assessment, the group that received elastase was an increase in the number of macrophages (22.88±2.24 cells/104um2), neutrophils (1.18±0.15 cells/104um2), positive TNF-alfa cells (12.52±0.42 cells/104um2) in the lung parenchyma. In remodeling changes in lung parenchyma, there was an increase in the volume ratio of collagen fibers (11.5 ± 0.11%), elastic (0.5±0.03%), the number of positive MMP-9 cells (18.59±1.87 cells/104um2), MMP-12 (20.17 ± 1.92 cells/104um2) TIMP-1 (14.42±2.05 cells/104um2) compared to controls (p < 0.001). Oxidative stress, was an increased of eNOS (13.15±0.40 cells/104um2), iNOS (10.49 ± 0.65 cells/104um2) and isoprostane (18.11±5.38%). Treatment CrataBL (ELA-CrataBL group) reduced the amount of parenchymal lung macrophages (9.58±1.36 cells/104um2), neutrophils (0.75±0.1 cells/104um2), positive TNF-alfa cells (10.4±0.49 cells/104um2), collagen (10.8±0.13%), elastic (0.3±0.02%), the number of positive MMP-9 cells (10.35±0.65 cells/104?m2), MMP-12 (14.15±0.59 cells/104um2), TIMP-1 (9.89±2.79 cells/104um2) MUC-5 (3.56±0.54 cells/104um2), eNOS (6.98±0:32 cells/104um2) and iNOS (6.21±0.42 cells/104um2) and isoprostane (8.96 ± 3.08%) compared to group ELA (p < 0.001). Airway was also a significant increase in neutrophils (5.97±1.03 cells/104um2), positive TNF-alfa cells (15.82±1.03 cells/104um2). Changes in lung airway remodeling also occurred an increase in the volume ratio of collagen fibers (8.73±2.59%), elastic (2.56±0.18%), the number of positive MMP-9 cells (14.86±1.77 cells/104um2), MMP-12 (18.56±1.79 cells/104um2) TIMP-1 (1.31±0.12 cells/104um2) and MUC-5 (7.09±1.71 cells/104um2) compared to controls (p < 0.001). Oxidative stress, an increase of eNOS (3.09 ± 0.08 cells/104um2), iNOS (5.4±0.3 cells/104um2) and isoprostane (18.11±5.38%) compared to controls (p < 0.001). Treatment CrataBL (ELA-CrataBL group) reduced the amount airway neutrophils (4.62±0.61 cells/104um2), TNF-alfa (14.30 ± 1.28 cells/104um2), collagen fibers (7 80±1.37%), elastic (1.4±0.13%), the number of positive MMP-9 cells (9.93±1.39 cells/104um2), MMP-12 (12.06±1.15), TIMP-1 (0.73±0.05 cells/104um2), MUC-5 (3.56±0.54 cells/104um2), eNOS (1.89±0,16 cells/104um2) and iNOS (4.3±0.31 cells/104um2), isoprostane (7.34±2.31%) compared to group ELA (p < 0.001). Conclusion: CrataBL attenuates changes in lung mechanics, broncho alveolar inflammatory responsiveness, control remodeling and oxidative stress induced by elastase. Although more studies should be conducted, this bifunctional protein may contribute as a potential therapeutic tool for the treatment of COPD

O objetivo deste trabalho foi avaliar se a proteína bifuncional de planta, CrataBL, que tem lectina e as propriedades inibidoras de enzima, modula alterações de mecânica pulmonar, inflamatórias e remodelamento induzidas por elastase intratraqueal em camundongos. Métodos: 36 camundongos C57BL6 receberam elastase (0,025 mg) por instilação intratraqueal (grupo ELA e ELA-CrataBL). Os grupos controles receberam salina (grupo SAL e SAL-CrataBL). Os camundongos foram tratados com instilação intraperitoneal de CrataBL (2mg/kg) nos dias 1, 14 e 21 após a instilação intratraqueal de elastase (grupo SAL-CrataBL e ELA-CrataBL) os animais controle receberam salina no mesmo volume. No dia 28, os camundongos foram anestesiados, ventilados mecanicamente e foram analisados a resistência e elastância do sistema respiratório (Ers e Rrs), elastância e resistência tecidual (Htis e Gtis), resistência das vias aéreas (Raw) e óxido nítrico exalado (NOex). Após, o lavado broncoalveolar (LBA) foi realizado, os pulmões foram retirados e por morfometria, e foram quantificados o intercepto linear médio (Lm), a quantidade de neutrófilos, células positivas para TNF-alfa, fibras colágenas, elásticas, células positivas para MMP-9, MMP-12, TIMP-1, eNOS e iNOS e isoprostano no parênquima pulmonar e vias aéreas. No parênquima foram avaliados os macrófagos nos septos alveolares e nas vias aéreas, foram também avaliadas as células para MUC-5. Resultados: No grupo ELA houve um aumento na Ers, Raw, Gtis, Htis, Lm, NOex, nas células totais, macrófagos, neutrófilos, eosinófilos e linfócitos no LBA em relação aos controles (p < 0,05), sendo que Raw, diminuiu também nos grupos SAL-CrataBL e ELA-CrataBL. Nos grupos tratados com CrataBL houve uma diminuição de Ers (37,0±2,2 cmH2O/L), Htis (37,9±3,5 cmH2O/ml/s), ENO (14,7±0,7 ppb), comparativamente ao grupo ELA (p < 0,05). No LBA houve atenuação de neutrófilos (0,003±0,001 104células/ml), linfócitos (0,003±0,001 104células/ml) e de Lm (54,6±6,0 mm). Complementando a avaliação, no grupo que recebeu elastase houve um aumento no número de macrófagos (22,88 +- 2,24 células/104um2), neutrófilos (1,18 +- 0,15 células/10 4um2), células positivas para TNF-ala (12,52 +- 0,42 células/104um2) no parênquima pulmonar. Nas alterações de remodelamento no parênquima pulmonar, houve um aumento da proporção de volume de fibras colágenas (11,5 +- 0,11%), elásticas (0,5 +- 0,03%), na quantidade de células positivas para MMP-9 (18,59 +- 1,87 células/104?m2), MMP-12 (20,17 +- 1,92 células/104?m2), TIMP-1 (14,42 +- 2,05 células/104um2) em comparação com os controlos (p < 0,001). No estresse oxidativo, houve um aumento de eNOS (13,15 +- 0,40 células/104um2), iNOS (10,49 +- 0,65 células/104um2) e isoprostano (18,11 =- 5,38%). O tratamento CrataBL (grupo ELA-CrataBL) reduziu no parênquima pulmonar a quantidade de macrófagos (9,58 +- 1,36 células/104um2), neutrófilos (0,75 +- 0,1 células/104um2), células positivas para TNF-alfa (10.4±0,49 células/104?m2), fibras colágenas (10,8 +- 0,13%), elásticas (0,3 +- 0,02%), a quantidade de células positivas para a MMP-9 (10,35±0,65 células/104um2), MMP-12 (14,15±0,59 células/104um2), TIMP-1 (9,89 +- 2,79 células/104um2), MUC-5 (3,56 +- 0,54 células/104um2), eNOS (6.98 +- 0.32 células/104um2) e iNOS (6,21 +- 0,42 células/104um2) e isoprostano (8,96 +- 3,08 %) em relação ao grupo ELA (p < 0,001). Nas vias aéreas também ocorreu um aumento significativo de neutrófilos (5,97 +- 1,03 células/104um2), células positivas para TNF-alfa (15,82 +- 1,03 células/104um2). Nas alterações de remodelamento pulmonar nas vias aéreas também ocorreu um aumento da proporção de volume de fibras colágenas (8,73 +- 2,59%), elásticas (2,56 +- 0,18%), na quantidade de células positivas para MMP-9 (14,86 +- 1,77 células/104um2), MMP-12 (18,56 +- 1,79 células/104um2), TIMP-1 (1,31 +- 0,12 células/104um2) e MUC-5 (7,09 +- 1,71 células/104um2) em comparação com os controlos (p < 0,001). No estresse oxidativo, houve um aumento de células positivas para eNOS (3,09 +- 0,08 células/104um2), iNOS (5,4 +- 0,3 células/104um2) e isoprostano (18,11 +- 5,38%) em comparação com os controlos (p < 0,001). O tratamento CrataBL (grupo ELA-CrataBL) reduziu nas vias aéreas a quantidade de neutrófilos (4,62 +- 0,61 células/104um2), TNF- alfa (14,30 +- 1,28 células/104um2), fibras colágenas (7,80 +- 1,37%), elásticas (1,4 +- 0,13%), a quantidade de células positivas para a MMP-9 (9,93 +- 1,39 células/104um2), MMP-12 (12,06 +- 1,15 células/104um2), TIMP-1 (0,73 +- 0,05 células/104?m2), MUC-5 (3,56 +- 0,54 células/104um2), eNOS (1,89 +- 0,16 células/104um2) e iNOS (4,3 +- 0,31 células/104um2), isoprostano (7,34 +- 2,31%) em relação ao grupo ELA (p < 0,001). Conclusão: CrataBL atenua as alterações de mecânica pulmonar, lavado bronco alveolar, responsividade inflamatória, controle do remodelamento e estresse oxidativo induzidas pela elastase. Embora mais estudos devam ser realizados, esta proteína bifuncional pode contribuir como potencial ferramenta terapêutica para o tratamento da DPOC / The aim of this study was to evaluate whether the bifunctional protein plant, CrataBL, which has lectin and enzyme inhibitory properties, modulates changes in lung mechanics, inflammatory and remodeling induced by intratracheal elastase in mice.Methods : 36 C57/Bl6 mice received elastase (0.025 mg) by intratracheal (group ELA and ELA-CrataBL). Control groups received saline (group SAL and SAL-CrataBL).The mice were treated with intraperitoneal instillation of CrataBL (2mg/kg) on days 1, 14 and 21 after intratracheal instillation of elastase (group SAL-CrataBL and ELA-CrataBL), control animals received saline in the same volume. On day 28, the mice were anesthetized and mechanically ventilated were analyzed resistance and respiratory system elastance (Ers and Rrs), elastance and tissue resistance (Htis and Gtis), airway resistance (Raw) and exhaled nitric oxide (ENO). After the bronchoalveolar lavage (BAL) was performed, the lungs were removed and morphometry were quantified and the linear intercept mean (Lm), the number of neutrophils, positive cells for TNF-alfa, collagen fibers, positive cells for MMP-9, MMP-12, TIMP-1, eNOS, iNOS and isoprostane in lung parenchyma and airways. Parenchyma was also evaluated macrophages in the alveolar septa. Airway was also evaluated MUC-5 cells. Results: In group ELA was an increase in Ers, Raw, Gtis, Htis, Lm, ENO, in total cells, macrophages, neutrophils, eosinophils and lymphocytes in BAL compared to controls (p < 0.05), and Raw, decreased in both groups SAL-CrataBL and ELA-CrataBL. In the groups treated with CrataBL there was a decrease in Ers (37.0±2.2 cmH2O/L) Htis (37 9±3.5 cmH2O/ml/s) and ENO (14.7±0.7 ppb) compared to the ELA group (p < 0.05). In BAL there was attenuation of neutrophils (0.003±0.001 104cells/ml), lymphocytes (0.003±0.001 104cells/ml) and Lm (54.6±6.0 mm). Complementing the assessment, the group that received elastase was an increase in the number of macrophages (22.88±2.24 cells/104um2), neutrophils (1.18±0.15 cells/104um2), positive TNF-alfa cells (12.52±0.42 cells/104um2) in the lung parenchyma. In remodeling changes in lung parenchyma, there was an increase in the volume ratio of collagen fibers (11.5 ± 0.11%), elastic (0.5±0.03%), the number of positive MMP-9 cells (18.59±1.87 cells/104um2), MMP-12 (20.17 ± 1.92 cells/104um2) TIMP-1 (14.42±2.05 cells/104um2) compared to controls (p < 0.001). Oxidative stress, was an increased of eNOS (13.15±0.40 cells/104um2), iNOS (10.49 ± 0.65 cells/104um2) and isoprostane (18.11±5.38%). Treatment CrataBL (ELA-CrataBL group) reduced the amount of parenchymal lung macrophages (9.58±1.36 cells/104um2), neutrophils (0.75±0.1 cells/104um2), positive TNF-alfa cells (10.4±0.49 cells/104um2), collagen (10.8±0.13%), elastic (0.3±0.02%), the number of positive MMP-9 cells (10.35±0.65 cells/104?m2), MMP-12 (14.15±0.59 cells/104um2), TIMP-1 (9.89±2.79 cells/104um2) MUC-5 (3.56±0.54 cells/104um2), eNOS (6.98±0:32 cells/104um2) and iNOS (6.21±0.42 cells/104um2) and isoprostane (8.96 ± 3.08%) compared to group ELA (p < 0.001). Airway was also a significant increase in neutrophils (5.97±1.03 cells/104um2), positive TNF-alfa cells (15.82±1.03 cells/104um2). Changes in lung airway remodeling also occurred an increase in the volume ratio of collagen fibers (8.73±2.59%), elastic (2.56±0.18%), the number of positive MMP-9 cells (14.86±1.77 cells/104um2), MMP-12 (18.56±1.79 cells/104um2) TIMP-1 (1.31±0.12 cells/104um2) and MUC-5 (7.09±1.71 cells/104um2) compared to controls (p < 0.001). Oxidative stress, an increase of eNOS (3.09 ± 0.08 cells/104um2), iNOS (5.4±0.3 cells/104um2) and isoprostane (18.11±5.38%) compared to controls (p < 0.001). Treatment CrataBL (ELA-CrataBL group) reduced the amount airway neutrophils (4.62±0.61 cells/104um2), TNF-alfa (14.30 ± 1.28 cells/104um2), collagen fibers (7 80±1.37%), elastic (1.4±0.13%), the number of positive MMP-9 cells (9.93±1.39 cells/104um2), MMP-12 (12.06±1.15), TIMP-1 (0.73±0.05 cells/104um2), MUC-5 (3.56±0.54 cells/104um2), eNOS (1.89±0,16 cells/104um2) and iNOS (4.3±0.31 cells/104um2), isoprostane (7.34±2.31%) compared to group ELA (p < 0.001). Conclusion: CrataBL attenuates changes in lung mechanics, broncho alveolar inflammatory responsiveness, control remodeling and oxidative stress induced by elastase. Although more studies should be conducted, this bifunctional protein may contribute as a potential therapeutic tool for the treatment of COPD

Submitted by Alberto Vieira (martins_vieira@ibest.com.br) on 2018-01-12T17:42:34Z No. of bitstreams: 1 576675.pdf: 24206483 bytes, checksum: f9816b609327b2737e0382acc3943b53 (MD5) / Made available in DSpace on 2018-01-12T17:42:34Z (GMT). No. of bitstreams: 1 576675.pdf: 24206483 bytes, checksum: f9816b609327b2737e0382acc3943b53 (MD5) Previous issue date: 2002 / Estuda a biologia floral, a fenologia e a reprodução de Capparis lineata Domb. ex Pers., Capparis flexuosa (L.) L. e Cleome rosea Vahl. ex DC., em áreas de restinga, localizadas em Itaipuaçu e Maricá (RJ, Brasil), no período de 1997 a 2000. As duas espécies de Capparis apresentam flores brancas, planas, nectaríferas, perfumadas, hermafroditas e noturnas. As flores são polinizadas por esfingídios, fato confirmado pelas observações de campo e pela presença de escamas de lepidópteros noturnos sobre os estigmas. Capparis lineata é uma trepadeira pouco frequente na área de estudos, apresenta floração anual e sincrônica que se estende de setembro (final da estação fria e seca) a dezembro (meados da estação quente e chuvosa), com pico em outubro. Capparis flexuosa tem hábito variando de arbusto prostrado a arvoreta e ocorre desde a zona da praia até a fimbria da mata, sendo muito frequente nas comunidades densas e fechadas que ocorrem sobre os cordões arenosos. Apresenta floração extensa, durante cerca de 10 meses do ano, com vários episódios de emissão floral, os quais são mais sincrônicos nos meses de dezembro/janeiro e março (estação quente e chuvosa), quando as flores duram apenas uma noite e originam muitos frutos com muitas sementes. Nos episódios que ocorrem na estação fria e seca, as flores permanecem atrativas pela manhã, podendo ser polinizadas por Xylocopa ordinaria, originando, entretanto, frutos pequenos e com poucas sementes. São feitas considerações acerca do envolvimento de formigas na dispersão das sementes de C. flexuosa. O estudo mostra uma grande variabilidade morfológica entre todas as espécies esfingófilas encontradas na área de estudo, além de uma forte sazonalidade do evento de floração das mesmas, que ocorre, predominantemente, na estação quente e chuvosa. Capparis lineata e C. flexuosa são auto-incompatíveis, com proporções Fruto/Flor de 6,9% e 45%, respectivamente. A baixa produção de frutos em C. lineata foi investigada, tendo-se concluído que tanto a limitação de polinizadores quanto a falta de recursos energéticos maternos estão envolvidos no número de frutos produzidos por planta. Cleome rosea é monocárpica, com ciclo de vida anual. As populações naturais e cultivadas apresentam sistema sexual subdióico, com plantas exclusivamente femininas, as quais emitem apenas flores pistiladas, e plantas poliníferas, que emitem flores hermafroditas (7,9% a 48,7%), estaminadas (50% a 92,1 %) e pistiladas (0 a 2,4%). As flores são zigomorfas, de cor rosa, nectaríferas e polinizadas, principalmente, por borboletas. Flores hermafroditas apresentam hercogamia, que previne a auto-polinização espontânea. Nas plantas poliníferas, as flores hermafroditas, em geral, são emitidas na base das inflorescências e antecedem a emissão das flores estaminadas, caracterizando uma dicogamia inter-floral. São apresentadas evidências de que a presença de frutos na base das inflorescências inibe a emissão de novas flores hermafroditas e acelera o início da fase de emissão de flores estaminadas. A espécie é autocompatível. As sementes produzidas pelas plantas femininas são resultantes de xenogamia, enquanto que aquelas produzidas pelas plantas poliníferas podem ser resultantes de geitonogamia. Em condições naturais, as plantas femininas produzem mais frutos e sementes do que as plantas poliníferas. Os frutos das primeiras contêm sementes com menor índice de aborto e com maior taxa de germinação. / Studies floral biology, phenology and reproductive system of Capparis lineata Domb. ex Pers., Capparis flexuosa (L.) L. and Cleome rosea Vahl. ex DC., at sandy coastal plains ("restingas") of Maricá (RJ, Brasil), from 1997 to 2000. The two species of Capparis present white, dish-type, nectariferous, scented, hermaphrodite and nocturnal flowers. The pollination by sphingids had been confirmed based on observation of natural populations and by the finding of nocturnal lepidoptera's scales over the stigmas. Capparis lineata is a climber uncommon in the study area. It presents annual and synchronous flowering which remains from September (end of the cold and dry season) to December (rniddle of the hot and rainy season), with its peak in October. Capparis flexuosa varies from a prostrate shrub to a small tree and is found from the vicinity of the beach to the edge of the woods, being more common in the dense scrub communities at the sandy layers. It presents extensive flowering, during ten months in the year, with several episodes of floral emission, which are more synchronous in December, January and March (hot and rainy season), when the flowers last only one night, producing many fruits with many seeds. During the cold and dry season, the flowers remain attractive in the morning, and can be pollinated by Xylocopa ordinaria, generating, however, smaller fruits with fewer seeds. Considerations are risen about the participation of ants in the seed dispersion of C. flexuosa. The study shows a great morphological variability between the flowers of all the sphingophyllous species found at the study area, and also a strong seasonality of the flowering periods, tending towards the wet and hot season. Capparis lineata e C. jlexuosa are self-incompatible, with Fruit/Flower proportions of 6.9% and 45%, respectively. The possible causes of the low fruit-set of C. lineata has been investigated and evidences of pollinator limitation and selective abortion of fruits is presented. Cleome rosea is an annual monocarpic species. The natural and cultivated populations present female plants, with only pistilate flowers, and polliniferous plants, with hermaphrodite (7.9% to 48.7%), staminate (50% to 92.1%) and pistilate flowers (0 to 2.4%), characterizing a subdioecious sexual system. The zygomorphic, pink, nectariferous flowers of C. rosea are mainly pollinated by butterflies. The hennaphrodites flowers have intra-floral hercogamy, which prevent the spontaneous self-pollination, and, in general, are produced at the base of the inflorescences and precede the emission of the staminate flowers, characterizing an inter-floral dicogamy. Evidences are showed that the presence of developing fruits at the base of the inflorescences inhibit the new hemaphrodite flowers and accelerate the beginning of the staminate phase of flower emission. The species is self-compatible. The seeds produced by the female plants are always cross-fertilized, while the seeds produced by the polliniferous plants may be self-fertilized by geitonogamy. In natural conditions, the female plants produce more fruits and seeds, with smaller rate of abortion and greater rate of germination than the polliniferous ones.