The Effects of High-Fat Diet and Flaxseed Oil-Enriched Diet on the Lung Parenchyma of Obese Mice.

Omega-3 (ω3) fatty acids are widely investigated for their anti-inflammatory potential, however, there is little evidence regarding their action in the lung parenchyma in the context of obesity. The objective is to investigate the effects of flaxseed oil (FS), rich in α-linolenic (C18:3 - ω3), on the lungs of obese mice. Mice were fed a high-fat diet (HF) for 8 weeks to induce obesity. Subsequently, a part of these animals received HF containing FS oil for another 8 weeks. The HF consumption induced weight gain and hyperglycemia. The lung parenchyma shows a complete fatty acids profile, compared to the control group (CT). In the lung parenchyma, FS increases the ω3 content and, notwithstanding a reduction in the interleukins (IL) IL1ß and IL18 contents compared to HF. However, FS promoted increased alveolar spaces, followed by MCP1 (Monocytes Chemoattractant Protein-1) positive cell infiltration and a dramatic reduction in the anti-inflammatory cytokine, IL10. Despite reducing the pulmonary inflammatory response, the consumption of a food source of ω3 was associated with alterations in the lipid profile and histoarchitecture of the lung parenchyma, which can lead to the development of pulmonary complications. This study brings an alert against the indiscriminate use of ω3 supplements, warranting caution.

Antiproliferative and Pro-Oxidant Effect of Polyphenols in Aqueous Leaf Extract of Passiflora alata Curtis on Activated T Lymphocytes from Non-Obese Diabetic (NOD SHILT/J) Mice.

The antioxidant, anti-inflammatory and antiproliferative properties of Passiflora alata Curtis are due to the presence of polyphenols in its composition. Our previous work showed that non-obese diabetic (NOD) mice undergoing treatment with aqueous leaf extract of P. alata present reduced insulitis in the pancreas, possibly due to its anti-inflammatory properties. However, depending on the concentration and their ability to interact with other molecules, these phenolic compounds may promote oxidation reactions in some cellular components, such as proteins and lipids, thus presenting a pro-oxidant effect. The present work aimed to evaluate the in vitro effects of aqueous leaf extract of P. alata and its polyphenols (vitexin, isoorientin, rutin and catechin) on lymphocyte proliferation and viability, the cell cycle and oxidative stress. Our results showed that T lymphocytes stimulated with concanavalin A mitogen (ConA) and in the presence of IC50 concentrations of P. alata extract and polyphenols undergo cell injury via inhibition of proliferation, with these effects being more pronounced concerning CD4+ T cells (P. alata, 3.54 ± 0.34%; isoorientin, 57.07 ± 6.4%; vitexin, 16.95 ± 1.11%; catechin, 37.9 ± 4.2% and rutin, 40.14 ± 4.5%), compared to the non-treated group (77.17 ± 6.29) (p < 0.0001 for all comparisons). This process includes late apoptosis/necrosis induction (P. alata, 77.5 ± 0.7%; vitexin, 83 ± 3.3%; isoorientin, 83.8 ± 1.4%; catechin, 83 ± 1.9% and rutin, 74.9 ± 3.2, while the control presented 53.6% ± 3.1 (p < 0.0001 for all comparisons)) and mitochondrial depolarization leading to cell-death induction. Furthermore, an in vitro model of a mixed culture of NOD mice T cells with a mouse pancreatic beta-cell line (MIN6) showed increased intracellular nitric oxide and lipid peroxidation in NOD T cells submitted to P. alata extract (46.41 ± 3.08) compared to the untreated control group (33.57 ± 1.99, p = 0.01315). These results suggest that aqueous leaf extract of P. alata and the polyphenols in these leaves represent a target for translational research showing the plant's benefits for developing new drugs with immunomodulatory properties against inflammatory diseases such as diabetes mellitus.

De Novo Membranous Nephropathy Associated With Antibody-Mediated Rejection in Kidney Transplant Recipients.

BACKGROUND: Membranous nephropathy (MN) is a rare autoimmune disease that can develop a persistent nephrotic syndrome and end-stage kidney disease, with a recurrence rate of 30% to 40% after kidney transplant. METHODS: Retrospective case series of membranous nephropathy observed in a cohort of kidney transplant recipients with donor-specific anti-human leukocyte antigen antibodies and biopsy-proven antibody-mediated rejection (AMR). RESULTS: We report 4 cases of membranous nephropathy associated with AMR. MN was diagnosed 10 to 92 months posttransplant, associated with de novo donor-specific antibodies, specific to class I in 2 cases, and class II in another 2. All cases presented typical morphology of membranous nephropathy, with subepithelial deposits with spikes at electron microscopy. Immunostaining for immunoglobulin G4 was negative in all cases, and podocyte-expressed M-type phospholipase A2 receptor was detected in glomerular basement membrane of 3 cases. Biopsy specimens from patients with longer follow-up showed more intense microvascular inflammation and chronic injury markers, possibly because of subclinical immunologic injury. AMR therapy included immunoglobulin 2g/kg in 3 patients, isolated or associated with plasmapheresis. One patient was not treated because of an active disseminated infection. Two patients remain with functioning grafts and under antiproteinuric therapy. Two grafts were lost, 1 because of chronic failure and the other because of death secondary to infection. Despite treatment, donor-specific antibodies remain detectable in a 6-month follow-up. CONCLUSIONS: De novo MN is a rare manifestation associated with AMR in kidney transplant recipients. The occurrence of podocyte-expressed M-type phospholipase A2 receptor in de novo MN suggests antibody-mediated activation, despite the use of maintenance immunosuppression.

Biological and Inflammatory Effects of Antigen 5 from Polybia paulista (Hymenoptera, Vespidae) Venom in Mouse Intraperitoneal Macrophages.

The social wasp Polybia paulista (Hymenoptera, Vespidae) is highly aggressive, being responsible for many medical occurrences. One of the most allergenic components of this venom is Antigen 5 (Poly p 5). The possible modulation of the in vitro immune response induced by antigen 5 from P. paulista venom, expressed recombinantly (rPoly p 5), on BALB/c mice peritoneal macrophages, activated or not with LPS, was assessed. Here, we analyzed cell viability changes, expression of the phosphorylated form of p65 NF-κB subunit, nitric oxide (NO), proinflammatory cytokines production, and co-stimulatory molecules (CD80, CD86). The results suggest that rPoly p 5 does not affect NO production nor the expression of co-stimulatory molecules in mouse peritoneal macrophages. On the other hand, rPoly p 5 induced an increase in IL-1ß production in non-activated macrophages and a reduction in the production of TNF-α and MCP-1 cytokines in activated macrophages. rPoly p 5 decreased the in vitro production of the phosphorylated p65 NF-κB subunit in non-activated macrophages. These findings suggest an essential role of this allergen in the polarization of functional M2 macrophage phenotypes, when analyzed in previously activated macrophages. Further investigations, mainly in in vivo studies, should be conducted to elucidate Polybia paulista Ag5 biological role in the macrophage functional profile modulation.

Renal transplant patients with preformed anti-HLA antibodies: early biopsy findings and clinical outcomes.

INTRODUCTION: Renal fibrosis is the end point of a process that begins at transplant, with ischemia reperfusion and early inflammation, and progresses over time with immunological and non-immunological phenomena. Early identification of morphological markers and intervention could improve graft function and survival. OBJECTIVE: to evaluate the correlation between intensity and specificity of pre-transplant anti-HLA antibodies and kidney allograft pathology in order to identify early risk factors or markers of allograft dysfunction. METHODS: A retrospective cohort of kidney transplant recipients with pre-transplant anti-HLA antibodies who underwent graft biopsy within the first two years post-transplant was divided into two groups according to the specificity of anti-HLA antibodies: nonspecific (non-DSA, n = 29) and specific (DSA+, n = 16). Kidney graft pathology, renal function, and proteinuria were analyzed. RESULTS: general characteristics were similar in both groups, except for the higher dose of thymoglobulin in DSA+ group (p < 0.05). The non-DSA group had higher scores for glomerulosclerosis, interstitial inflammation (i) and interstitial fibrosis (ci) (p < 0.05) and higher incidence of cell-mediated acute rejection. No statistical difference in incidence of antibody-mediated rejection, renal function, and proteinuria was observed during follow up. DISCUSSION AND CONCLUSIONS: the difference in inflammation scores and interstitial fibrosis may be associated to the higher incidence of acute cell-mediated rejection and polyomavirus nephropathy in the Non-DSA group. We also should take into account the protective effect of higher doses of thymoglobulin, reducing ischemia reperfusion injury in the DSA+ group. The short follow-up might have been insufficient to detect long-term changes in allograft tissue, renal function, and proteinuria.

Identification and Antioxidant Activity of the Extracts of Eugenia uniflora Leaves. Characterization of the Anti-Inflammatory Properties of Aqueous Extract on Diabetes Expression in an Experimental Model of Spontaneous Type 1 Diabetes (NOD Mice).

Medical and folklore reports suggest that Eugenia uniflora (E. uniflora) is a functional food that contains numerous compounds in its composition, with anti-inflammatory, antioxidant and anti-diabetic effects. In the present study, we investigated the best solvents (water, ethanol and methanol/acetone) for extracting bioactive compounds of E. uniflora leaves, assessing total phenols and the antioxidant activity of the extracts by 2,2-Diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), 2,2'-Azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and Oxygen Radical Absorbance Capacity (ORAC) assays, identifying hydrolysable tannins and three phenolic compounds (ellagic acid, gallic acid and rutin) present in the leaves. In addition, we evaluated the incidence of diabetes, degree of insulitis, serum insulin, hepatic glutathione and tolerance test glucose in non-obese diabetic (NOD) mice. Our results suggest that the aqueous extract presents antioxidant activity and high total phenols, which were used as a type 1 diabetes mellitus (DM-1) treatment in NOD mice. We verified that the chronic consumption of aqueous extract reduces the inflammatory infiltrate index in pancreatic islets, maintaining serum insulin levels and hepatic glutathione, and reducing serum lipid peroxidation as well as the risk for diabetes.

Capacidade antioxidante e composição química da casca de maracujá (Passiflora edulis) / Antioxidant capacity and chemical composition of passion fruit peel (Passiflora edulis)

O objetivo do estudo foi avaliar a composição centesimal da farinha obtida da casca do maracujá e sua atividade antioxidante in vitro, buscando viabilizar suas potencialidades para incorporação em produtos alimentares. Os frutos foram higienizados, as cascas foram separadas da polpa, secas em estufa com circulação de ar e moídas para obtenção de uma farinha. Foi determinada a composição centesimal e os polifenóis presentes na casca foram extraídos por três metodologias, tendo sido quantificados o teor de fenóis totais e a atividade antioxidante por três métodos distintos (sequestro do radical 2,2-difenil-1-picrilhidrazil - DPPH, capacidade antioxidante de redução do ferro - FRAP e capacidade de absorção do radical oxigênio - ORAC). Os resultados mostram que a farinha apresenta elevado teor de fibras (65,22±0,27%), das quais 74% são fibras insolúveis. Embora os resultados do teor de polifenóis indiquem que possivelmente há um predomínio de compostos com características hidrofílicas na casca, visto que a água foi o solvente que apresentou maior rendimento na extração dos fenóis totais, a avaliação da atividade antioxidante dos extratos mostra que o poder antioxidante dos compostos lipofílicos presentes no extrato metanólico/acetona foram superiores ao dos compostos presentes no extrato aquoso e etanólico. Os resultados obtidos no presente estudo demonstram que a casca do maracujá poderia ser utilizada como fonte de fibras e antioxidantes. No entanto, sua incorporação em formulações alimentares deve atentar as suas características físicas, químicas e sensoriais, para garantir seu melhor aproveitamento e aceitação entre os consumidores. .

The aim of the study was to evaluate the chemical composition of flour obtained from the peel of passion fruit and their antioxidant activity in vitro. The fruits were cleaned, the peels were separated from pulp, dried in an oven with circulating air and ground to obtain the flour. The centesimal composition was determined and the polyphenols present in the peel were extracted by three methodologies. Total phenolic content and antioxidant activity were determined using three different methods (scavenging of 2,2-diphenyl-1-picrylhydrazyl - DPPH free radical, Ferric Reducing Antioxidant Power - FRAP e Oxigen Radical Absorbance Capacity - ORAC). The results show that the flour has high fiber content (65.22±0.27%) and 74% of which correspond to insoluble fraction. Although the results of total polyphenol indicate that there is a predominance of compounds with hydrophilic characteristics in the peel, the evaluation of the antioxidant activity of the extracts showed that the antioxidant power of lipophilic compounds present in the methanol / acetone extract was superior to the compounds present in the aqueous and ethanol extracts. The results of this study demonstrate that the passion fruit peel could be used as a source of fiber and antioxidants, however , its incorporation into food formulations must address their physical, chemical and sensory, to ensure their best use and acceptance among consumers. .

In search of a tolerance-induction strategy for cow's milk allergies: significant reduction of beta-lactoglobulin allergenicity via transglutaminase/cysteine polymerization.

OBJECTIVE: To explore the use of ß-lactoglobulin polymerized using microbial transglutaminase and heating to identify whether protein polymerization could reduce in vivo allergenicity and maintain in vitro and ex vivo immunoreactivity for use in tolerance-induction protocols. METHODS: Based on previous protocols applied in mice and children, we performed in vivo challenges (using a skin prick test) with native and polymerized ß-lactoglobulin in adult patients with an IgE-mediated allergy to plactoglobulin. In vitro humoral immunoreactivity was analyzed using immunoblotting. Cell-mediated immunoreactivity was analyzed using ex vivo challenges with native and polymerized ß-lactoglobulin and monitored by leukocyte adherence inhibition tests. RESULTS: The skin tests demonstrated that there was a significant reduction in immediate cutaneous reactivity after polymerization. Polymerization did not decrease the immunoblotting detection of s-IgE specific to ß-lactoglobulin. Cell-mediated immunoreactivity, as assessed by ex vivo challenges and leukocyte adherence inhibition tests, did not exhibit significant differences between leukocytes challenged with native versus polymerized ß-lactoglobulin. CONCLUSIONS: The polymerization of ß-lactoglobulin decreased in vivo allergenicity and did not decrease in vitro humoral or ex vivo cell-mediated immunoreactivity. Therefore, we conclude that inducing polymerization using transglutaminase represents a promising technique to produce suitable molecules for the purpose of designing oral/ sublingual tolerance induction protocols for the treatment of allergies.

In search of a tolerance-induction strategy for cow's milk allergies: significant reduction of beta-lactoglobulin allergenicity via transglutaminase/cysteine polymerization

OBJECTIVE: To explore the use of β-lactoglobulin polymerized using microbial transglutaminase and heating to identify whether protein polymerization could reduce in vivo allergenicity and maintain in vitro and ex vivo immunoreactivity for use in tolerance-induction protocols. METHODS: Based on previous protocols applied in mice and children, we performed in vivo challenges (using a skin prick test) with native and polymerized β-lactoglobulin in adult patients with an IgE-mediated allergy to plactoglobulin. In vitro humoral immunoreactivity was analyzed using immunoblotting. Cell-mediated immunoreactivity was analyzed using ex vivo challenges with native and polymerized β-lactoglobulin and monitored by leukocyte adherence inhibition tests. RESULTS: The skin tests demonstrated that there was a significant reduction in immediate cutaneous reactivity after polymerization. Polymerization did not decrease the immunoblotting detection of s-IgE specific to β-lactoglobulin. Cell-mediated immunoreactivity, as assessed by ex vivo challenges and leukocyte adherence inhibition tests, did not exhibit significant differences between leukocytes challenged with native versus polymerized β-lactoglobulin. CONCLUSIONS: The polymerization of β-lactoglobulin decreased in vivo allergenicity and did not decrease in vitro humoral or ex vivo cell-mediated immunoreactivity. Therefore, we conclude that inducing polymerization using transglutaminase represents a promising technique to produce suitable molecules for the purpose of designing oral/ sublingual tolerance induction protocols for the treatment of allergies.

Diacerhein downregulate proinflammatory cytokines expression and decrease the autoimmune diabetes frequency in nonobese diabetic (NOD) mice.

NOD mice are used as experimental models as they develop type 1 diabetes mellitus (DM-1) spontaneously, with a strong similarity to the human disease. Diabetes mellitus type 1 is characterized by the destruction of the islet, orchestrated by T lymphocytes that induce cytokine release like IL-1beta, promoting an inflammatory process. Diacerhein has antiinflammatory properties, inhibiting IL-1. However, the mechanisms involved in immune modulation are not completely understood. In the present study, serum and pancreatic islets were isolated to investigate the relationship between IL-1beta, IFN-gamma, IL-12 and TNF-alpha expression and diabetes onset, morphological aspects, and diacerhein dose dependence in animals treated with different doses (5, 10 and 50 mg/kg/day) and the control group (saline solution). The results demonstrated upregulation of mRNA islets and downregulation of the serum concentration of IL-1beta, IL-12 and TNF-alpha in the group treated with 5 and 10 mg/kg/day diacerhein, when compared with the saline group, and increased IFN-gamma serum concentration in the group treated with 50 mg/kg/day. These results suggest that diacerhein in NOD mice, decreases, in a dose-dependent manner, the diabetes frequency downregulating proinflammatory cytokines, such as IL-1beta, TNF-alpha, IFN-gamma and IL-12 at posttranscriptional or posttranslational level. Furthermore, using the HPLC method, diacerhein and rhein (active metabolite) were detected in serum and pancreas of treated mice.

Perfil de auto-anticorpos na esclerose sistêmica / Autoantibodies in systemic sclerosis

Objetivo: Estudar a incidência de auto-anticorpos séricos numa populaçao de pacientes com esclerose sistêmica (ES),verificando sua importância clínica e analisando sua distribuiçao quanto às formas da doença. Métodos: Foram avaliados 58 pacientes com ES. O fator antinuclear (FAN) e o anticorpo anticentrômero foram pesquisados por imunofluorescência indireta, utilizando com substrato células HEp-2. Os anticorpos antitopoisomerase I, anti-RNP, anti-Sm, anti-SS-A/Ro e anti-SS-B/La foram pesquisados pela técnica de micro-hemaglutinaçao. Foram comparados com as formas clínicas da ES, difusa (ESd) e limitada (ESl), além de sexo, raça, escore cutâneo total (ECT), calcinose, telangiectasias e manifestaçoes viscerais. Resultados: O FAN mostrou-se positivo em 82//dos pacientes, nos padroes pontilhado, nucleolar e centromérico. O anticorpo anticentrômero foi positivo em 18 por cento dos casos, todos com ESl, estando associado ao ECT< ou = 20, à calcinose e às telangiectasias. O anticorpo antitopoisomerase I foi positivo em 25 por cento dos pacientes, estando associado ao ECT > 20 e aos acometimentos articular e vascular, independente da forma clínica. Nao houve predomínio de anticorpos anticentrômero ou antitopoisomerase I em relaçao às manifestaçoes viscerais. O anticorpo anti-RNP foi positivo em dois pacientes, o anti-SS-A/Ro em cinco, enquanto que o anti-Sm e o anti-SS-B/La foram negativos em todos os casos. Conclusoes: Os resultados obtidos confirmam a associaçao do anticorpo anticentrômero com a ESl e a presença do anticorpo antitopoisomerase I nas duas formas clínicas de ES, associado ao envolvimento cutâneo mais extenso

Detecçäo de anticorpos e anticomponentes lipídicos (cardiolipina, gangliosídios e galactocerebrosídios) em pacientes com doença de Behçet / Detection of antibodies and lipid anticomponents (cardiolipin, gangliosides and galactosilceramidas) in patients with Behçet's disease

Os autores avaliam a presença de anticorpos antifraçöes lipídicas (cardiolipina, gangliosídios e galactocerebrosídios) em 13 pacientes com doença de Behcet. Anticorpos séricos anticardiolipina do isotipo IgG foram detectados em 30// dos casos, 50// do isotipo IgM. Em nenhum dos pacientes foi observada presença de isotipo IgA. Antigalactocerebrosídio foram detectados em apenas dois (15//) dos pacientes. Com relaçäo às manifestaçöes clínicas, observou-se que a presença de anticorpos anticardiolipina configura fator de risco para o desenvolvimento do acometimento neurológico