2 nd Brazilian Consensus on Chagas Disease, 2015

Abstract Chagas disease is a neglected chronic condition with a high burden of morbidity and mortality. It has considerable psychological, social, and economic impacts. The disease represents a significant public health issue in Brazil, with different regional patterns. This document presents the evidence that resulted in the Brazilian Consensus on Chagas Disease. The objective was to review and standardize strategies for diagnosis, treatment, prevention, and control of Chagas disease in the country, based on the available scientific evidence. The consensus is based on the articulation and strategic contribution of renowned Brazilian experts with knowledge and experience on various aspects of the disease. It is the result of a close collaboration between the Brazilian Society of Tropical Medicine and the Ministry of Health. It is hoped that this document will strengthen the development of integrated actions against Chagas disease in the country, focusing on epidemiology, management, comprehensive care (including families and communities), communication, information, education, and research

Undernutrition affects cell survival, oxidative stress, Ca2+ handling and signaling pathways in vas deferens, crippling reproductive capacity.

BACKGROUND: The aim of this work was to investigate the mechanisms by which chronic malnutrition (CM) affects vas deferens function, leading to compromised reproductive capacity. Previous studies have shown that maternal malnutrition affects the reproductive tracts of adult male offspring. However, little is known about the effects of CM, a widespread life-long condition that persists from conception throughout growth to adult life. METHODOLOGY/PRINCIPAL FINDINGS: Young adult male rats, which were chronically malnourished from weaning, presented decreased total and haploid cells in the vas deferens, hypertrophy of the muscle layer in the epididymal portion of the vas deferens and intense atrophy of the muscular coat in its prostatic portion. At a molecular level, the vas deferens tissue of CM rats exhibited a huge rise in lipid peroxidation and protein carbonylation, evidence of an accentuated increase in local reactive oxygen species levels. The kinetics of plasma membrane Ca(2+)-ATPase activity and its kinase-mediated phosphorylation by PKA and PKC in the vas deferens revealed malnutrition-induced modifications in velocity, Ca(2+) affinity and regulation of Ca(2+) handling proteins. The severely crippled content of the 12-kDa FK506 binding protein, which controls passive Ca(2+) release from the sarco(endo) plasmic reticulum, revealed another target of malnutrition related to intracellular Ca(2+) handling, with a potential effect on forward propulsion of sperm cells. As a possible compensatory response, malnutrition led to enhanced sarco(endo) plasmic reticulum Ca(2+)-ATPase activity, possibly caused by stimulatory PKA-mediated phosphorylation. CONCLUSIONS/SIGNIFICANCE: The functional correlates of these cellular and molecular hallmarks of chronic malnutrition on the vas deferens were an accentuated reduction in fertility and fecundity

Probiotic Prato cheese attenuates cigarette smoke-induced injuries in mice

111

Estudo imunohistoquímico do remodelamento pulmonar em camundongos expostos à fumaça de cigarro Immunohistochemical study of lung remodeling in mice exposed to cigarette smoke

OBJETIVO: Muitos estudos sobre enfisema são realizados com exposição de animais à fumaça de cigarro durante um longo tempo, focando o tipo de célula envolvida no desequilíbrio protease/antiprotease e a degradação da matriz extracelular. A expressão aumentada de metaloproteinases no enfisema está associado com citocinas e evidências sugerem um papel importante da metaloproteinase de matriz-12 (MMP-12). Nosso objetivo foi estudar a detecção de inibidor tissular de metaloproteinase-2 (TIMP-2), fator de necrose tumoral alfa (TNF-&#945;) e interleucina-6 (IL-6) por métodos imunohistoquímicos no pulmão de camundongos. MÉTODOS: Camundongos C57BL/6 machos foram expostos 3 vezes ao dia a fumaça de 3 cigarros por um período de 10, 20, 30 ou 60 dias através de uma câmara de inalação (grupos CS10, CS20, CS30 e CS60, respectivamente). O grupo controle foi exposto às mesmas condições ao ar ambiente. RESULTADOS: Nós observamos um aumento progressivo de macrófagos alveolares no lavado broncoalveolar dos grupos expostos. O diâmetro alveolar médio, um indicador de destruição alveolar, aumentou em todos os grupos expostos quando comparado ao grupo controle. O índice imunohistoquímico (II) para MMP-12 aumentou nos grupos CS10, CS20 e CS30 em paralelo a uma redução do II para TIMP-2 nos grupos CS10, CS20 e CS30. O II para as citocinas TNF-&#945; e IL-6 aumentou em todos os grupos expostos quando comparado ao grupo controle. Enfisema foi observado no grupo CS60, com alterações na densidade de volume de fibras colágenas e elásticas. CONCLUSÕES: Estes achados sugerem que a fumaça de cigarro induz enfisema com uma participação importante do TNF-&#945; e da IL-6 sem a participação de neutrófilos.<br>OBJECTIVE: Various studies of emphysema involve long-term exposure of animals to cigarette smoke, focusing on the cell type involved in the protease/antiprotease imbalance and on extracellular matrix degradation. In emphysema, increased expression of metalloproteinases is associated with cytokines, and evidence suggests that the matrix metalloproteinase-12 (MMP-12) plays an important role. Our objective was to investigate tissue inhibitor of metalloproteinase-2 (TIMP-2), tumor necrosis factor-alpha (TNF-&#945;) and interleukin-6 (IL-6) detection by immunohistochemical methods in mouse lung. METHODS: Male C57BL/6 mice were exposed 3 times a day to smoke of 3 cigarettes over a period of 10, 20, 30 or 60 days in an inhalation chamber (groups CS10, CS20, CS30 and CS60, respectively). Controls were exposed to the same conditions in room air. RESULTS: A progressive increase in the number of alveolar macrophages was observed in the bronchoalveolar lavage fluid of the exposed mice. The mean linear intercept, an indicator of alveolar destruction, was greater in all exposed groups when compared to control group. In the CS10, CS20 and CS30 mice, the immunohistochemical index (II) for MMP-12 increased in parallel with a decrease in II for TIMP-2 in the CS10, CS20 and CS30 mice. The II for the cytokines TNF-&#945; and IL-6 was greater in all exposed groups than in the control group. Emphysema, with changes in volume density of collagen and elastic fibers, was observed in the CS60 group. CONCLUSIONS: These findings suggest that cigarette smoke induces emphysema with major participation of TNF-&#945; and IL-6 without participation of neutrophils

Effects of oleanolic acid on pulmonary morphofunctional and biochemical variables in experimental acute lung injury

We analysed the effects of oleanolic acid (OA) on lung mechanics and histology and its possible mechanisms of action in experimental acute lung injury (ALI). BALB/c mice were randomly divided into Control (saline, ip) and ALI (paraquat, 25 mg/kg, ip) groups. At 1 h, both groups were treated with saline (SAL, 50 \u3bcl ip), OA (10 mg/kg ip), or dexamethasone (DEXA, 1 mg/kg ip). At 24 h, lung static elastance, viscoelastic pressure, and alveolar collapse reduced more after OA compared to DEXA administration. Tumour necrosis factor-\u3b1, macrophage migration inhibitory factor, interleukin-6, interferon-\u3b3, and transforming growth factor-\u3b2 mRNA expressions in lung tissue diminished similarly after OA or DEXA. Conversely, only OA avoided reactive oxygen species generation and yielded a significant decrease in nitrite concentration. OA and DEXA restored the reduced glutathione/oxidized glutathione ratio and catalase activity while increasing glutathione peroxidase induced by paraquat. In conclusion, OA improved lung morphofunction by modulating the release of inflammatory mediators and oxidative stress

Ca²⁺-ATPase activity and content of Ca²⁺-handling proteins in whole homogenate of <i>vas</i><i>deferens</i> tissue.

<p>(A) Time course of Ca²⁺-ATPase activity. The Ca²⁺-ATPase activity was assayed at the different times of incubation presented on the <i>abscissa</i>, starting at 30 s, using the CTRL (empty circles) and CM (filled circles) groups. Results are expressed as mean ± SEM (n = 5). The smooth curves were fitted to the experimental points using the equation P_{i<i>, t</i>} = P_{i<i>, max</i>} (1 - e^{<i>-k t</i>}), where <i>k</i> is the first-order rate constant and P_{<i>i, max</i>} is the asymptotic value of P_i release. (B) and (C) Ca²⁺ concentration dependence of the Ca²⁺-ATPase activities in whole homogenate of <i>vas deferens</i>. Values were obtained for PMCA (B) and SERCA (C) activities of CTRL (empty circles) and CM (filled circles) groups. Results are expressed as mean ± SEM (n = 5). The smooth lines were adjusted to the experimental points by non-linear regression using the equations: v = V_max × [Ca²⁺]/(K_{0.5 Ca} + [Ca²⁺]) or v = v_c + V_max × [Ca²⁺]/(K_{0.5 Ca} + [Ca²⁺]) for the CTRL and CM groups, respectively. V_max is the maximal hydrolysis rate, and K_{0.5 Ca} is the free Ca²⁺ concentration required for half-maximal activation of the CTRL activities and also represents the free Ca²⁺ concentration required for half-maximal activation of the lower affinity components (above 0.01 µM) of PMCA and SERCA in the CM group. The term v_c represents the very high affinity component of the Ca²⁺ curve in CM. (D) (E) and (F) Contents of PMCA, SERCA2 and FKBP12 in <i>vas deferens</i>. Ca²⁺ pumps PMCA and SERCA2 and immunophilin FKBP12 were immunodetected in whole homogenates of <i>vas deferens</i> from the CTRL (empty bars) and CM (black bars) groups. (D) PMCA. Upper panel: representative immunoblotting of three determinations. Lower panel: graphic representation of three immunoanalyses of different preparations. Samples from CTRL and CM were analyzed in parallel on the same gel; densitometric values were corrected for protein loading using Ponceau red and are presented as percentage of CTRL. (E) SERCA2. Upper panel: representative immunoblotting of three determinations. Lower panel: graphic representation of three immunoanalyses of different preparations. Samples were analyzed as described in (D). (F) FKBP12. Upper panel: representative immunoblotting of three determinations from different rats from the CTRL and CM groups, each of which was analyzed using recombinant FKPB12 as a parallel positive control, as shown in immunoblotting legend. Lower panel: graphic representation of three immunoanalyses of <i>vas deferens</i> FKPB12. *P<0.05 <i>vs</i>. the corresponding CTRL; NS: no statistical difference.</p

Kinetic parameters: Ca²⁺ concentration dependence of PMCA and SERCA from CTRL and CM preparations of <i>vas deferens</i> (n = 5).

<p>¹ Parameters corresponding to the lower affinity component of the Ca²⁺ curve.</p