Effect of supplementation with extract of white bean flour in murine model

Common bean supplementation (Phaseolus vulgaris) "in natura" causes loss of body weight associated with a deficiency in nutrient absorption and histopathological changes. This effect has been attributed to phytohemagglutinin (PHA) present in high concentrations in red and white beans. The main objective of this work was to evaluate the safety of white bean flour as a dietary supplement. Animals were treated for 14 days with extract of white bean flour (WBFE) at doses of 2.65g/kg and 5.30g/kg. A significant reduction in body weight was observed, accompanied by the reduction of mean values of glycemia, in both groups in relation to the control group. Significant structural changes were also observed in the intestinal epithelium. Additionally, mice treated with WBFE 5.30g/kg presented mononuclear inflammatory infiltrate in the lamina propria of the intestinal mucosa accompanied by a dose-dependent increase in the dosage of chemokine MCP-1 and nitric oxide, although without causing intestinal and hepatic oxidative and oxidative damage.  The deleterious effects resulting from the use of the WBFE are not permanent since the treated animals after 14 days without WBFE stimuli. In conclusion, commercial bean flour did not prove to be safe as oral dietary supplementation at the dosages used because of the antinutritional and immunomodulatory effects.

Evaluation of the oxidative and inflammatory profile of patients with migraine

Among one of the most commonly seen and primary headaches is migraine. Oxidative stress and inflammation is discussed to be implicated in the pathogenesis of migraine. However, further knowledge about this issue is necessary because data are in part controversial and the possible underlying mechanisms remain inconclusive to date. To evaluate and compare the oxidative and inflammatory profile of patients with migraine 47 volunteers were divided into 3 groups: 15 women with chronic migraine (WCM); 17 women with episodic migraine (WEM) and 15 men with migraine (MM) and enrolled in this study.Total antioxidant status; the enzymatic activity of the antioxidant agents Catalase (CAT), Glutathione S-transferase (GST) and Superoxide Dismutase (SOD); oxidative stress markers malondialdehyde (MDA) and carbonylated proteins; blood count and neutrophil/lymphocyte ratio; uric acid, c-reactive protein and cholesterol total and fractions were determined in pacients with migraine in the interictal phase. The group male participants (MM) displayed a reduction in total antioxidant status, as well as a lower value for antioxidant enzymes, but had no significant alterations in markers related to damage by oxidative stress compared to women. These findings suggest that there is a difference in the oxidative profile between the sexes among migraine patients. This may allow a better understanding of patients profile with different migraine phenotypes and identify new markers that might help understanding the pathophysiology and migraine patient management

Evaluation of inflammatory and oxidative profile in patients with recurrent wheezing

Wheezing is a clinical expression of numerous respiratory-related conditions. Although asthma is the leading cause of wheezing during childhood. The present study aims to evaluate the inflammatory and oxidative profile of pediatric patients with recurrent wheezing. Seventy-eight volunteers were divided into three groups according to their age (≤ 36, 36 to 72 and ≥ 73 months). Blood was collected for hematological evaluation, serum detections of total immunoglobulin E (IgE), and C-reactive protein (PCRus). The oxidative profile was evaluated by total antioxidant capacity (FRAP), malondialdehyde (MDA) and carbonylated protein markers. There was no significant difference in the IgE and PCRus levels among the three groups evaluated. However, a significant positive correlation was observed for PCRus with total leukocyte and with neutrophils for the group of patients ≥ 73 months of age. The intermediate age group presented significantly reduced FRAP values in the serum, while significant values of oxidative damage markers were observed in the group of patients ≥ 73 months of age. When determining the correlation between inflammatory and oxidative markers, only the ≥ 73 months group showed significant. The group ≥ 73 months stands out with significant alterations of the oxidative stress markers and their correlations with the inflammatory profile

Identification of BOL lectin ligands and its role in cell signaling

A BOL é uma proteína do tipo TRAF com atividade de lectina, extraída de Brassica oleracea ssp. botrytis. Trabalhos anteriores identificaram a capacidade da BOL de induzir fagocitose, produção de espécies reativas de oxigênio em macrófagos. O objetivo deste estudo foi analisar a capacidade de ligação da BOL à superfície de leucócitos e a identificação dos seus possíveis ligantes. Demonstramos pela primeira vez que a BOL é capaz de ligar a superfície de linfócitos e macrófagos; capacidade esta que pode estar relacionada aos efeitos observados em macrófagos. Ainda, através do ensaio de MTT foi possível observar que a BOL é capaz de induzir proliferação em esplenócitos, sendo a concentração mínima de 1,25 µg/mL suficiente para indução. Através da cromatografia de afinidade por lectina, com uma resina acoplada com BOL, foi possível purificar 4 possíveis ligantes dessa lectina. O sequenciamento proteico por MALDI/TOF revelou quatro potenciais ligantes, TF3C2, NEST, ELNF1 e FERL3. Análises in silico predisseram os possíveis sítios de glicosilação dessas proteínas e acessibilidade de resíduos na superfície proteica, indicando que as proteínas possuem sítios de glicosilação acessíveis na sua superfície, confirmando a possibilidade desses possíveis ligantes serem glicoproteínas. Essas proteínas estão envolvidas em diferentes níveis com a sinalização celular, proliferação e citoesqueleto, desse modo, a interação da lectina BOL, contendo uma estrutura com apenas dois domínios MATH, pode indicar a possível atuação desse domínio como um CRD e o de lectinas em eventos de sinalização celular. Palavras-chave: Lectina. Domínios proteicos. Sinalização. Imunologia.BOL is a TRAF-protein with lectinic activity extracted from Brassica oleracea ssp. Botrytis. Previous work has identified BOL's ability to induce phagocytosis, production of reactive oxygen species in macrophages. The aim of this study was to analyze the capacity of BOL to bind to the leukocyte surface and to identify its possible ligands. We demonstrated for the first time that BOL is capable of binding the surface of lymphocytes and macrophages; capacity which may be related to the effects observed in macrophages. Through the MTT assay it was possible to observe that BOL is capable of inducing proliferation in splenocytes, with a minimum concentration of 1.25 µg/mL sufficient for induction. Through lectin affinity chromatography, with a BOL- coupled resin, it was possible to purify 4 possible ligands of this lectin. Protein sequencing by MALDI/TOF revealed four potential ligands, TF3C2, NEST, ELNF1, and FERL3. In silico analyzes predicted the possible glycosylation sites of these proteins and accessibility of residues on the protein surface, indicating that the proteins have accessible glycosylation sites on their surface, confirming the possibility of these possible ligands being glycoproteins. These proteins are involved at different levels with cell signaling, proliferation and cytoskeleton organization, so the interaction of lectin BOL, containing a structure with only two MATH domains, may indicate the possible role of this domain as a CRD and the role of lectins in cell signaling events. Keywords: Lectin. Protein domains. Signaling. Lymphocyte

Hallmarks of Aging in Macrophages: Consequences to Skin Inflammaging

The skin is our largest organ and the outermost protective barrier. Its aging reflects both intrinsic and extrinsic processes resulting from the constant insults it is exposed to. Aging in the skin is accompanied by specific epigenetic modifications, accumulation of senescent cells, reduced cellular proliferation/tissue renewal, altered extracellular matrix, and a proinflammatory environment favoring undesirable conditions, including disease onset. Macrophages (Mφ) are the most abundant immune cell type in the skin and comprise a group of heterogeneous and plastic cells that are key for skin homeostasis and host defense. However, they have also been implicated in orchestrating chronic inflammation during aging. Since Mφ are related to innate and adaptive immunity, it is possible that age-modified skin Mφ promote adaptive immunity exacerbation and exhaustion, favoring the emergence of proinflammatory pathologies, such as skin cancer. In this review, we will highlight recent findings pertaining to the effects of aging hallmarks over Mφ, supporting the recognition of such cell types as a driving force in skin inflammaging and age-related diseases. We will also present recent research targeting Mφ as potential therapeutic interventions in inflammatory skin disorders and cancer

Heterologous Expression, Purification, and Immunomodulatory Effects of Recombinant Lipoprotein GUDIV-103 Isolated from Ureaplasma diversum

Ureaplasma diversum is a bacterial pathogen that infects cattle and can cause severe inflammation of the genital and reproductive systems. Lipid-associated membrane proteins (LAMPs), including GUDIV-103, are the main virulence factors in this bacterium. In this study, we heterologously expressed recombinant GUDIV-103 (rGUDIV-103) in Escherichia coli, purified it, and evaluated its immunological reactivity and immunomodulatory effects in bovine peripheral blood mononuclear cells (PBMCs). Samples from rabbits inoculated with purified rGUDIV-103 were analysed using indirect enzyme-linked immunosorbent assay and dot blotting to confirm polyclonal antibody production and assess kinetics, respectively. The expression of this lipoprotein in field isolates was confirmed via Western blotting with anti-rGUDIV-103 serum and hydrophobic or hydrophilic proteins from 42 U. diversum strains. Moreover, the antibodies produced against the U. diversum ATCC 49783 strain recognised rGUDIV-103. The mitogenic potential of rGUDIV-103 was evaluated using a lymphoproliferation assay in 5(6)-carboxyfluorescein diacetate succinimidyl ester–labelled bovine PBMCs, where it induced lymphocyte proliferation. Quantitative polymerase chain reaction analysis revealed that the expression of interleukin-1β, toll-like receptor (TLR)-α, TLR2, TLR4, inducible nitric oxide synthase, and caspase-3–encoding genes increased more in rGUDIV-103–treated PBMCs than in untreated cells (p < 0.05). Treating PBMCs with rGUDIV-103 increased nitric oxide and hydrogen peroxide levels. The antigenic and immunogenic properties of rGUDIV-103 suggested its suitability for immunobiological application