O veneno de serpentes do gênero Bothrops (responsáveis por 80% dos envenenamentos no Brasil) é composto por metalo e serinoproteases, disintegrinas, fosfolipases, entre outros, e pode causar edema, hemorragia, necrose, e manifestações sistêmicas, como coagulação intravascular, choque, falência renal e hemorragia. O veneno da B. pirajai ativa o sistema complemento (C), sugerindo uma contribuição para o agravamento dos sintomas. Considerando a importância do C no processo inflamatório e o papel das metaloproteinases nos envenenamentos, verificou-se que o tratamento do sangue total humano (onde células e mediadores plasmáticos interagem entre si) com ou sem a compstatina (inibidor de C3 do C), e com a metaloproteinase de classe PI do veneno de B. pirajai, levou a diferenças bastante significativas na expressão dos marcadores analisados nos leucócitos, na geração de anafilatoxinas e TCC, e na quantificação de citocinas e quimiocinas no plasma, sugerindo que a inibição do C reduz o processo inflamatório, podendo ser uma terapia efetiva para envenenamentos botrópicos. / The snake venom of Bothrops genus (responsible for 80% of envenomation in Brazil) is composed by metallo and serine proteases, disintegrins, phospholipase, among others, and can cause edema, hemorrhage, necrosis, and systemic manifestations, such as intravascular coagulation, shock, renal failure and systemic hemorrhage. The venom of B. pirajai is able to activate the complement system (C), suggesting a contribution to the worsening of symptoms. Considering the importance of C in the inflammatory process and the role of metalloproteinases in envenomation, it was found that treatment of human whole blood (where cells and plasma mediators interact) with or without compstatin (C3 inhibitor), and with a class PI metalloproteinase from B. pirajai\'s venom led to highly significant differences in the expression of the markers analyzed in leukocytes, in generation of anaphylatoxins and TCC, and quantification of cytokines and chemokines in plasma, suggesting that inhibition of C reduces the inflammatory process and may be an effective therapy for bothropic envenomations.

<p>The aim of this study was to investigate the effects of Sutherlandia frutescens on the inflammatory response and T cell differentiation in vitro using cytokines as biomarkers. Whole blood cells containing various concentrations of Sutherlandia frutescens were stimulated in vitro with either Lipopolysaccharide (LPS) or Phytohaemagglutinin (PHA). Results show that Sutherlandia frutescens is not toxic at any of the concentrations tested. The addition of Sutherlandia frutescens at high concentrations to the stimulated whole blood cell cultures reflects a significant down regulation of Interleukin(IL) 6 and IL-10 compared to the control (P&lt / 0.05) hence suppressed the inflammatory and humoral immune response. Results obtained for Inteferon-gamma (IFN ) shows that Sutherlandia frutescens is donor specific as it reflects both up and down regulation in the release of IFN at the concentrations tested. The in vitro data generated by this study supports the use of Sutherlandia frutescens in the management of inflammatory conditions and allergies such as asthma. However the effects of Sutherlandia frutescens on cell mediated immunity was found to be donor specific. Further investigation of Sutherlandia frutescens on cellular immunity is advised.</p>

<p>The aim of this study was to investigate the effects of Sutherlandia frutescens on the inflammatory response and T cell differentiation in vitro using cytokines as biomarkers. Whole blood cells containing various concentrations of Sutherlandia frutescens were stimulated in vitro with either Lipopolysaccharide (LPS) or Phytohaemagglutinin (PHA). Results show that Sutherlandia frutescens is not toxic at any of the concentrations tested. The addition of Sutherlandia frutescens at high concentrations to the stimulated whole blood cell cultures reflects a significant down regulation of Interleukin(IL) 6 and IL-10 compared to the control (P&lt / 0.05) hence suppressed the inflammatory and humoral immune response. Results obtained for Inteferon-gamma (IFN ) shows that Sutherlandia frutescens is donor specific as it reflects both up and down regulation in the release of IFN at the concentrations tested. The in vitro data generated by this study supports the use of Sutherlandia frutescens in the management of inflammatory conditions and allergies such as asthma. However the effects of Sutherlandia frutescens on cell mediated immunity was found to be donor specific. Further investigation of Sutherlandia frutescens on cellular immunity is advised.</p>

O veneno de serpentes do gênero Bothrops (responsáveis por 80% dos envenenamentos no Brasil) é composto por metalo e serinoproteases, disintegrinas, fosfolipases, entre outros, e pode causar edema, hemorragia, necrose, e manifestações sistêmicas, como coagulação intravascular, choque, falência renal e hemorragia. O veneno da B. pirajai ativa o sistema complemento (C), sugerindo uma contribuição para o agravamento dos sintomas. Considerando a importância do C no processo inflamatório e o papel das metaloproteinases nos envenenamentos, verificou-se que o tratamento do sangue total humano (onde células e mediadores plasmáticos interagem entre si) com ou sem a compstatina (inibidor de C3 do C), e com a metaloproteinase de classe PI do veneno de B. pirajai, levou a diferenças bastante significativas na expressão dos marcadores analisados nos leucócitos, na geração de anafilatoxinas e TCC, e na quantificação de citocinas e quimiocinas no plasma, sugerindo que a inibição do C reduz o processo inflamatório, podendo ser uma terapia efetiva para envenenamentos botrópicos. / The snake venom of Bothrops genus (responsible for 80% of envenomation in Brazil) is composed by metallo and serine proteases, disintegrins, phospholipase, among others, and can cause edema, hemorrhage, necrosis, and systemic manifestations, such as intravascular coagulation, shock, renal failure and systemic hemorrhage. The venom of B. pirajai is able to activate the complement system (C), suggesting a contribution to the worsening of symptoms. Considering the importance of C in the inflammatory process and the role of metalloproteinases in envenomation, it was found that treatment of human whole blood (where cells and plasma mediators interact) with or without compstatin (C3 inhibitor), and with a class PI metalloproteinase from B. pirajai\'s venom led to highly significant differences in the expression of the markers analyzed in leukocytes, in generation of anaphylatoxins and TCC, and quantification of cytokines and chemokines in plasma, suggesting that inhibition of C reduces the inflammatory process and may be an effective therapy for bothropic envenomations.

Magister Scientiae - MSc / The aim of this study was to investigate the effects of Sutherlandia frutescens on the inflammatory response and T cell differentiation in vitro using cytokines as biomarkers. Whole blood cells containing various concentrations of Sutherlandia frutescens were stimulated in vitro with either Lipopolysaccharide (LPS) or Phytohaemagglutinin (PHA). Results show that Sutherlandia frutescens is not toxic at any of the concentrations tested. The addition of Sutherlandia frutescens at high concentrations to the stimulated whole blood cell cultures reflects a significant down regulation of Interleukin(IL) 6 and IL-10 compared to the control (P&lt;0.05) hence suppressed the inflammatory and humoral immune response. Results obtained for Inteferon-gamma (IFN ) shows that Sutherlandia frutescens is donor specific as it reflects both up and down regulation in the release of IFN at the concentrations tested. The in vitro data generated by this study supports the use of Sutherlandia frutescens in the management of inflammatory conditions and allergies such as asthma. However the effects of Sutherlandia frutescens on cell mediated immunity was found to be donor specific. Further investigation of Sutherlandia frutescens on cellular immunity is advised. / South Africa

O agregado proteico P-MAPA apresentou potencial imunomodulatório em diversos estudos, mas a sua ação sobre o sistema complemento e receptores do tipo Toll (TLRs) não é, ainda, conhecida. Neste estudo o P-MAPA promoveu ativação das vias clássica e alternativa do sistema complemento e produção de C3a e C5a. Utilizando um modelo ex vivo de sangue total humano, o composto promoveu aumento da expressão de CD11b e CD14, diminuição da expressão de C5aR, TLR2 e TLR4, em leucócitos de sangue periférico e também quando combinado com LPS, porém não promoveu alterações na expressão de C3aR. O P-MAPA induziu redução de IFN-g no plasma, aumento da produção de TNF-&alpha;, IL-8, IL-12 e peróxinitrito, mas não induziu produção de superóxido, IL-6, IL-1&beta;, TGF-&beta; ou IL-10. Por meio de testes in vivo, foi possível determinar a dose letal do P-MAPA. Em conjunto, os dados obtidos mostram que o P-MAPA apresenta ação pró-inflamatória em modelo ex vivo de sangue total humano e que o tratamento combinado com LPS leva a uma amplificação dos seus efeitos. / P-MAPA, a protein aggregate has been described as a promising immunomodulator, however, its role on the complement system and Toll-like receptors (TLRs) is unknown. In the study, P-MAPA has promoted activation of the complement\'s classical and alternative pathways and the production of C3a and C5a. Using an ex vivo model of human whole blood, the compound promoted increase of CD11b and CD14 expression, decrease of C5aR, TLR2 and TLR4, in peripheral blood leucocytes and when combined with LPS, but did not change C3aR expression. P-MAPA promoted reduction of IFN-g in plasma, increased production of TNF-&alpha;, IL-8, IL-12 and peroxynitrite, but did not induce the production of superoxide, IL-6, IL-1&beta;, TGF-&beta; or IL-10. Through in vivo tests, we were able to determine a lethal dose for P-MAPA. Altogether, our data indicate that P-MAPA has proinflammatory action in ex vivo model of human whole blood and that the treatment combined with LPS leads to amplification of its effects.

O sangue é uma suspensão de células contidas num líquido complexo chamado plasma. O termo sangue total refere-se a amostras de sangue com a totalidade de seus constituintes, parte sólida e líquida. Sendo os elementos químicos responsáveis por funções essenciais, como regulação osmótica, frequência cardíaca e contratibilidade, coagulação sanguínea e excitabilidade neuromuscular. A determinação de elementos químicos em fluidos corporais como sangue, soro, plasma, tecido e urina é usada como monitor de parte ou de todo o organismo. Nesse trabalho, utilizou-se a técnica de fluorescência de raios X (EDXRF) para análise de amostras de sangue total humano e animal, hamsters da espécie dourada (Mesocricetus auratus) e cavalos da raça crioula (Equus caballus). Nas amostras de sangue humano, foram determinados intervalos de referência de Na (1788 - 1826 &mu;g g-1), Mg (63 - 75 &mu;g g-1), P (602 - 676 &mu;g g-1), S (1519 - 1718 &mu;g g-1), Cl (2743 - 2867 &mu;g g-1), K (1508 - 1630 &mu;g g-1), Ca (214 - 228 &mu;g g-1), Cu (4 - 6 &mu;g g-1) e Zn (1 - 3 &mu;g g-1). Foram determinados intervalos de referência de Na (1714 - 1819 &mu;g g-1), Mg (51 - 79 &mu;g g-1), P (970 - 1080 &mu;g g-1), S (1231 - 1739 &mu;g g-1), Cl (2775 - 2865 &mu;g g-1), K (1968 - 2248 &mu;g g-1), Ca (209 - 257 &mu;g g-1), Cu (4 - 6 &mu;g g-1) e Zn (3 - 5 &mu;g g-1) para amostras de sangue de hamster dourado. As amostras de sangue de cavalo da raça crioula apresentaram os intervalos de: Na (1955 - 2013 &mu;g g-1), Mg (51 - 75 &mu;g g-1), P (443 - 476 &mu;g g-1), S (1038 - 1140 &mu;g g-1), Cl (2388 - 2574 &mu;g g-1), K (1678 - 1753 &mu;g g-1), Ca (202 - 213 &mu;g g-1), Cu (4,1 - 4,5 &mu;g g-1) e Zn (2,0 - 2,2 &mu;g g-1). Estudo comparativo dos resultados entre as técnicas de análise por ativação neutrônica (NAA) e EDXRF indica a igualdade de desempenho das técnicas analíticas na análise de matrizes biológicas. Os resultados contribuem no estabelecimento de intervalos de referência para a população brasileira saudável e para as referidas espécies de animais. / Blood is a suspension of cells contained in a complex liquid called plasma. The term whole blood refers to samples with both solid and liquid parts. Inorganic elements are responsible for essential functions, such as osmotic regulation, cardiac frequency and contractibility, blood clotting and neuromuscular excitability. The determination of inorganic elements in corporeal fluids such as blood, serum, plasma, tissue and urine is used as a monitor for a part or the whole organism. In this work, the X-Ray fluorescence technique (EDXRF) was used for the determination of inorganic elements in whole blood samples from humans and animals (golden hamsters, Mesocricetus auratus and crioula breed horses, Equus caballus). The reference intervals of Na (1788 - 1826 &mu;g g-1), Mg (63 - 75 &mu;g g-1), P (602 - 676 &mu;g g-1), S (1519 - 1718 &mu;g g-1), Cl (2743 - 2867 &mu;g g-1), K (1508 - 1630 &mu;g g-1), Ca (214 - 228 &mu;g g-1), Cu (4 -6 &mu;g g-1) e Zn (1 - 3 &mu;g g-1) were determined for human blood. The reference intervals, for golden hamster blood were found to be: Na (1714 - 1819 &mu;g g-1), Mg (51 - 79 &mu;g g-1), P (970 - 1080 &mu;g g-1), S (1231 - 1739 &mu;g g-1), Cl (2775 - 2865 &mu;g g-1), K (1968 - 2248 &mu;g g-1), Ca (209 - 257 &mu;g g-1), Cu (4 - 6 &mu;g g-1) e Zn (3 -5 &mu;g g-1). The reference intervals, for crioula breed horse blood, showed to be: Na (1955 - 2013 &mu;g g-1), Mg (51 - 75 &mu;g g-1), P (443 - 476 &mu;g g-1), S (1038 - 1140 &mu;g g-1), Cl (2388 - 2574 &mu;g g-1), K (1678 - 1753 &mu;g g-1), Ca (202 - 213 &mu;g g-1), Cu (4,1 - 4,5 &mu;g g-1) e Zn (2,0 - 2,2 &mu;g g-1). Comparative study between NAA and EDXRF, both techniques showed the same performance for the analyses of biological matrices. The results contribute for the establishment of reference intervals for the Brazilian healthy population and the referred animal species.

<p>This study shows that herbs can be effectively screened for potiential bio-activity using in vitro methods. Further studies will be needed to better explore Artemisia afra&rsquo / s effect on immunoregulation, particularly long term effects of the herb on the immune system and its effect on other disease states.</p>

<p>This study shows that herbs can be effectively screened for potiential bio-activity using in vitro methods. Further studies will be needed to better explore Artemisia afra&rsquo / s effect on immunoregulation, particularly long term effects of the herb on the immune system and its effect on other disease states.</p>

Magister Scientiae - MSc (Medical BioSciences) / In the past three decades there has been a huge increase in the use of herbal medicine globally. The active principles of these herbal medicines are mostly unknown with supportive evidence for safety and efficacy very rare. SeptilinTM is a phytopharmaceutical formulation which is recommended for the treatment and management of various infections. It has been claimed to have immunomodulatory actions that potentiates the body's immune response. The immunomodulatory activity of SeptilinTM has not been well investigated via appropriate in vitro models. Therefore this study was undertaken to investigate the in vitro effects of SeptilinTM on biomarkers of specific immune pathways by using WBC. Stimulated and unstimulated WBC were incubated with the product. Enzyme linked immunosorbent assays were used to screen for IL-6, IL-10, and IFN? as biomarkers for inflammation, humoral immunity, and cell mediated immunity, respectively. Results show that the presence of SeptilinTM in LPS stimulated WBC has no effect on the release of IL-6 and IFN? production but stimulated IL-10 production. SeptilinTM in unstimulated WBC has no effect on the release of IL-10 and IFN? production but stimulatory effects on IL-6 production.