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-α, IL-8, IL-12 e peróxinitrito, mas não induziu produção de superóxido, IL-6, IL-1β, TGF-β 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-α, IL-8, IL-12 and peroxynitrite, but did not induce the production of superoxide, IL-6, IL-1β, TGF-β 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.

Magister Scientiae - MSc / 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’s effect on immunoregulation, particularly long term effects of the herb on the immune system and its effect on other disease states. / South Africa

Magister Scientiae - MSc / "The aim of the study was to use in vitro human whole blood cultures to screen the water samples collected from the Eerste/Plankenbrug river system for cytotoxicity and inflammatory activity and for the first time investigate the impact on the cell- mediated and humoral immune pathways. Water samples were collected fronm the sites during the dry summer season and rainy winter season. Blood was collected from the healthy male volunteers and diluted with RPMI 1640. For cytotoxicity and inflammatory activity 2.5ul of blood for 18-20 hrs at 37 C... This study shows that waster from the Plankenbrug River is heavily polluted by contaminants from both the agricultural area and informal settlement of Kayamandi. These contaminants can be potentially immunotoxic during the summer season and they can result in inflammatory diarrheal disease and immunosuppression in exposed individuals..."

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 μg g-1), Mg (63 - 75 μg g-1), P (602 - 676 μg g-1), S (1519 - 1718 μg g-1), Cl (2743 - 2867 μg g-1), K (1508 - 1630 μg g-1), Ca (214 - 228 μg g-1), Cu (4 - 6 μg g-1) e Zn (1 - 3 μg g-1). Foram determinados intervalos de referência de Na (1714 - 1819 μg g-1), Mg (51 - 79 μg g-1), P (970 - 1080 μg g-1), S (1231 - 1739 μg g-1), Cl (2775 - 2865 μg g-1), K (1968 - 2248 μg g-1), Ca (209 - 257 μg g-1), Cu (4 - 6 μg g-1) e Zn (3 - 5 μg 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 μg g-1), Mg (51 - 75 μg g-1), P (443 - 476 μg g-1), S (1038 - 1140 μg g-1), Cl (2388 - 2574 μg g-1), K (1678 - 1753 μg g-1), Ca (202 - 213 μg g-1), Cu (4,1 - 4,5 μg g-1) e Zn (2,0 - 2,2 μg 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 μg g-1), Mg (63 - 75 μg g-1), P (602 - 676 μg g-1), S (1519 - 1718 μg g-1), Cl (2743 - 2867 μg g-1), K (1508 - 1630 μg g-1), Ca (214 - 228 μg g-1), Cu (4 -6 μg g-1) e Zn (1 - 3 μg g-1) were determined for human blood. The reference intervals, for golden hamster blood were found to be: Na (1714 - 1819 μg g-1), Mg (51 - 79 μg g-1), P (970 - 1080 μg g-1), S (1231 - 1739 μg g-1), Cl (2775 - 2865 μg g-1), K (1968 - 2248 μg g-1), Ca (209 - 257 μg g-1), Cu (4 - 6 μg g-1) e Zn (3 -5 μg g-1). The reference intervals, for crioula breed horse blood, showed to be: Na (1955 - 2013 μg g-1), Mg (51 - 75 μg g-1), P (443 - 476 μg g-1), S (1038 - 1140 μg g-1), Cl (2388 - 2574 μg g-1), K (1678 - 1753 μg g-1), Ca (202 - 213 μg g-1), Cu (4,1 - 4,5 μg g-1) e Zn (2,0 - 2,2 μg 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.

For the past few decades, the popularity of graphene oxide (GO) nanomaterials (NMs) has increased exceedingly due to their biomedical applications in drug delivery of anti-cancer drugs. Their unique physicochemical properties such as high surface area and good surface chemistry with unbound surface functional groups (e.g. hydroxyl - OH, carboxyl /ketone C=O, epoxy/alkoxy C-O, aromatic group C=C, etc) which enable covalent bonding with organic molecules (e.g. RNA, DNA) make GO NMs as excellent candidates in drug delivery nanocarriers. Despite the overwhelming biomedical applications, there are concerns about their genotoxicity on human DNA. Published genotoxicity studies on GO NMs were performed using non-commercial GO with 2-3 layers of GO sheets, synthesized in various laboratories with the potential for inter-laboratory variabilities. However, what has not been studied before is the effects of the commercial GO (15-20 sheets; 4-10% edge-oxidized; 1 mg/mL) in vitro on DNA damage in human whole blood and peripheral blood lymphocytes (PBL) from real-life patients diagnosed with chronic pulmonary diseases [asthma, chronic obstructive pulmonary disease (COPD), and lung cancer], and genotoxic endpoints compared with those from healthy control individuals to determine whether there are any differences in GO sensitivity. Thus, in the present study, we had characterized GO NMs using Zetasizer Nano for Dynamic Light Scattering (DLS) and zeta potential (ZP) in the aqueous solution, and electron microscopy using the Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) in the dry state, respectively. Cytotoxicity studies were conducted on human PBL from healthy individuals and patients (asthma, COPD, and lung cancer) using the Methylthiazolyldiphenyl-tetrazolium bromide (MTT) and Neutral Red Uptake (NRU) assays, respectively. The genotoxicity (DNA damage) and cytogenetic effects (chromosome aberration parameters) induced by GO NMs on human whole blood from healthy individuals and patients were studied using the Alkaline Comet Assay and Cytokinesis-blocked Micronucleus (CBMN) assay, respectively. Our results showed concentration-dependent increases in cytotoxicity, genotoxicity, and chromosome aberrations, with blood samples from COPD and lung cancer patients being more sensitive to DNA damage insults compared with asthma patients and healthy control individuals. Furthermore, the relative gene and protein expressions of TP53, CDKN1A/p21, and BCL-2 relative to GAPDH on human PBL were studied using the Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) and Western Blot techniques, respectively. Our results have shown altered gene and protein expression levels. Specifically, GO-induced cytotoxicity, genotoxicity, and micronuclei aberrations were associated with TP53 upregulation - a biomarker of DNA damage - in both patients and healthy individuals. These effects show that GO NMs have promising roles in drug delivery applications when formulated to deliver drug payload to COPD and cancer cells. However, the fact that cytotoxicity, genotoxicity, chromosome instability, and gene/protein expressions - biomarkers of cancer risk - were observed in healthy individuals are of concern to public health, especially in occupational exposures at micro levels at the workplace.