Effect of chronic supplementation with shark liver oil on immune responses of exercise-trained rats

Previous studies have reported that chronic supplementation with shark liver oil (SLO) improves immune response of lymphocyte, macrophage and neutrophil in animal models and humans. In a similar manner, exercise training also stimulates the immune system. However, we are not aware of any study about the association of exercise and SLO supplementation on immune response. Thus, our main goal was to investigate the effect of chronic supplementation with SLO on immune responses of exercise-trained rats. Male Wistar rats were divided into four groups: sedentary with no supplementation (SED, n = 20), sedentary with SLO supplementation (SEDslo, n = 20), exercised (EX, n = 17) and exercised supplemented with SLO (EXslo, n = 19). Rats swam for 6 weeks, 1.5 h/day, in water at 32 +/- A 1A degrees C, with a load of 6.0% body weight attached to the thorax of rat. Animals were killed 48 h after the last exercise session. SLO supplementation did not change phagocytosis, lysosomal volume, superoxide anion and hydrogen peroxide production by peritoneal macrophages and blood neutrophils. Thymus and spleen lymphocyte proliferation were significantly higher in SEDslo, EX, and EXslo groups compared with SED group (P < 0.05). Gut-associated lymphocyte proliferation, on the other hand, was similar between the four experimental groups. Our findings show that SLO and EX indeed are able to increase lymphocyte proliferation, but their association did not induce further stimulation in the adaptive immune response and also did not modify innate immunity.

Acridone alkaloids as potent inhibitors of cathepsin V

Cathepsin V is a lysosomal cysteine peptidase highly expressed in thymus, testis and corneal epithelium. Eleven acridone alkaloids were isolated from Swinglea glutinosa (Bl.) Merr. (Rutaceae), with eight of them being identified as potent and reversible inhibitors of cathepsin V (IC(50) values ranging from 1.2 to 3.9 mu M). Detailed mechanistic characterization of the effects of these compounds on the cathepsin V-catalyzed reaction showed clear competitive inhibition with respect to substrate, with dissociation constants (K(i)) in the low micromolar range (2, K(i) = 1.2 mu M; 6, K(i) = 1.0 mu M; 7, K(i) = 0.2 mu M; and 11, K(i) = 1.7 mu M). Molecular modeling studies provided important insight into the structural basis for binding affinity and enzyme inhibition. Experimental and computational approaches, including biological evaluation, mode of action assessment and modeling studies were successfully employed in the discovery of a small series of acridone alkaloid derivatives as competitive inhibitors of catV. The most potent inhibitor (7) has a K(i) value of 200 nM. (C) 2011 Elsevier Ltd. All rights reserved.

Biflavonoids from Araucaria angustifolia protect against DNA UV-Induced damage

Ultraviolet radiation is one of the most deleterious forms of radiation to terrestrial organisms and is involved in formation of mutagenic pyrimidine dimers and oxidized nucleotides. The biflavonoid fraction (BFF), extracted from needles of Araucaria angustifolia was capable of protecting calf thymus DNA from damage induced by UV radiation. This occurred through prevention of cyclobutane thymine dimer and 8-oxo-7,8-dihydro-2`-deoxyguanosine formation, this being quantified by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) in a multiple reaction monitoring mode (MRM) and by HPLC-coulometric detection, respectively. (C) 2009 Elsevier Ltd. All rights reserved.

Probing the binding of tetraplatinum(pyridyl)porphyrin complexes to DNA by means of surface plasmon resonance

Tetrapyridylporphyrins containing four chloro(2,2`-bipyridine)platinum(II) complexes attached at the meta (3-H(2)TPtPyP) and para (4-H(2)TPtPyP) positions of the peripheral pyridine ligands were synthesized and their interaction with DNA investigated. The compounds were isolated in the solid state and characterized by means of spectroscopic and analytical techniques. According to molecular simulations, the two isomers exhibit contrasting structural characteristics, consistent with a saddle shape configuration for 3-H(2)TPtPyP and a planar geometry for 4-H(2)TPtPyP. Surface plasmon resonance studies were carried out on the interaction of the complexes with calf thymus DNA, revealing a preferential binding of 3-H(2)TPtPyP, presumably at the DNA major grooves. (C) 2008 Elsevier Inc. All rights reserved.

Mutagenicity and DNA adduct formation of PAH, nitro-PAH, and oxy-PAH fractions of atmospheric particulate matter from Sao Paulo, Brazil

Urban particulate matter (UPM) contributes to lung cancer incidence. Here, we have studied the mutagenic activity and DNA adduct-forming ability of fractionated UPM extractable organic matter (EOM). UPM was collected with a high-volume sampler in June 2004 at two sites, one at street level adjacent to a roadway and the other inside a park within the urban area of the city of Sao Paulo, Brazil. UPM was extracted using dichloromethane, and the resulting EOM was separated by HPLC to obtain PAH, nitro-PAH, and oxy-PAH fractions which were tested for mutagenicity with the Salmonella strains TA98 and YG1041 with and without S9 metabolic activation. The PAH fraction from both sites showed negligible mutagenic activity in both strains. The highest mutagenic activity was found for the nitro-PAH fraction using YG1041 without metabolic activation; however, results were comparable for both sites. The nitro-PAH and oxy-PAH fractions were incubated with calf thymus DNA under reductive conditions appropriate for the activation of nitro aromatic compounds, then DNA adduct patterns and levels were determined with thin-layer chromatography (TLC) (32)p-postlabeling method using two enrichment procedures-nuclease PI digestion and butanol extraction. Reductively activated fractions from both sites produced diagonal radioactive zones (DRZ) of putative aromatic DNA adducts on thin layer plates with both enrichment procedures. No such DRZ were observed in control experiments using fractions from unexposed filters or from incubations without activating system. Total adduct levels produced by the nitro-PAH fractions were similar for both sites ranging from 30 to 45 adducts per 10(8) normal nucleotides. In contrast, the DNA binding of reductively activated oxy-PAH fractions was three times higher and the adduct pattern consisted of multiple discrete spots along the diagonal line on the thin layer plates. However, DNA adduct levels were not significantly different between the sampling sites. Both samples presented the same levels of mutagenic activity. The response in the Salmonella assay was typical of nitroaromatics. Although, more mutagenic activity was related to the nitro-PAH fraction in the Salmonella assay, the oxy-PAH fractions showed the highest DNA adduct levels. More studies are needed to elucidate the nature of the genotoxicants occurring in Sao Paulo atmospheric samples. (C) 2008 Elsevier B.V. All rights reserved.

Larvicidal activity of the water extract of Moringa oleifera seeds against Aedes aegypti and its toxicity upon laboratory animals

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Diferentes dietas e pesos ao abate na produção de órgãos de cordeiros

O experimento foi conduzido com o objetivo de avaliar os pesos e porcentagens de órgãos em relação ao peso vivo ao abate (PVA), ao peso de corpo vazio (PCV) e ao peso total dos órgãos (PTO) de cordeiros inteiros 1/2 Ile de France 1/2 Ideal confinados, recebendo dietas com relações volumoso:concentrado de 50:50 e 30:70 e abatidos aos 30 ou 34 kg. Os parâmetros mensurados foram: língua, pulmões com traquéia, coração, fígado, pâncreas, timo, tireóide, adrenal, rins, testículos, baço, diafragma, bexiga, pênis e glândulas anexas. Os resultados obtidos revelaram diferenças entre os pesos ao abate nas porcentagens de língua, coração, pulmões com traquéia, diafragma, pâncreas, tireóide, baço e glândulas anexas em relação ao PVA; nas porcentagens de língua, pulmões com traquéia, fígado, pâncreas, baço e glândulas anexas em relação ao PCV e nas porcentagens de pulmões com traquéia, fígado, pâncreas, tireóide, baço e glândulas anexas em relação ao PTO. Dietas com diferentes relações volumoso:concentrado afetaram apenas as porcentagens de pâncreas em relação ao PVA, ao PCV e ao PTO. Verificaram-se interações entre relação volumoso:concentrado e peso vivo ao abate nos pesos dos pulmões com traquéia, fígado, pâncreas, tireóide e glândulas anexas.

Chronic restraint or variable stresses differently affect the behavior, corticosterone secretion and body weight in rats

Organisms are constantly subjected to stressful stimuli that affect numerous physiological processes and activate the hypothalamo-pituitary-adrenal (HPA) axis, increasing the release of glucocorticoids. Exposure to chronic stress is known to alter basic mechanisms of the stress response. The purpose of the present study was to compare the effect of two different stress paradigms (chronic restraint or variable stress) on behavioral and corticosterone release to a subsequent exposure to stressors. Considering that the HPA axis might respond differently when it is challenged with a novel or a familiar stressor we investigated the changes in the corticosterone levels following the exposure to two stressors: restraint (familiar stress) or forced novelty (novel stress). The changes in the behavioral response were evaluated by measuring the locomotor response to a novel environment. In addition, we examined changes in body, adrenals, and thymus weights in response to the chronic paradigms. Our results showed that exposure to chronic variable stress increased basal plasma corticosterone levels and that both, chronic restraint and variable stresses, promote higher corticosterone levels in response to a novel environment, but not to a challenge restraint stress, as compared to the control (non-stressed) group. Exposure to chronic restraint leads to increased novelty-induced locomotor activity. Furthermore, only the exposure to variable stress reduced body weights. In conclusion, the present results provide additional evidence on how chronic stress affects the organism physiology and point to the importance of the chronic paradigm and challenge stress on the behavioral and hormonal adaptations induced by chronic stress. (c) 2006 Elsevier B.V. All rights reserved.

Behavioral and neuroendocrine effects of the exposure to chronic restraint or variable stress in early adolescent rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Thymic lymphomas in Wistar rats exposed to N-methyl-N-nitrosourea (MNU)

The Brazilian Agency for the Environment (IBAMA) recently adopted an alternative medium-term multiple-organ assay system with the Wistar rat strain for detection of the carcinogenic potential of pesticides. Originally, this initiation-promotion protocol was established in Japan with the isogenic Fischer 344 male rat. Among the initiating agents used in that assay, N-methyl-N-nitrosourea (MNU) rapidly induces malignant lymphoma and leukemia and early mortality of rats from different strains. This study was developed to evaluate whether the outbred Wistar rats are also similarly susceptible to MNU. Particularly, it aimed to evaluate the dose-response relationship and to register the MNUinduced pre-neoplasia and neoplasia that may develop in the lympho-hematopoietic system (LHS) of the Wistar rat within a medium-term period. Four groups of male Wistar rats were treated during 2 weeks with vehicle or with MNU (80, 160 or 240 mg/kg body weight, i.p.). After sacrifice at the 12th and 20th weeks, the thymus, spleen, bone marrow, cervical and mesenteric lymph nodes and liver were collected for analysis. At the 20th week, LHS malignant tumors and benign vascular tumors occurred only in the high- and intermediate-dose MNU-treated animals. Four animals treated with 240 mg/kg developed diffuse thymic lymphomas; two others, treated respectively with 240 mg/kg and 160 mg/kg, developed spleen hemangiomas. The present observations indicate that the Wistar strain is as susceptible as other strains to the early development of MNU-induced LHS (pre)neoplasia. Therefore, this strain seems suitable to be used as test system in bioassay protocols that adopt MNU as an initiating agent for carcinogenesis.

Chemically induced immunotoxicity in a medium-term multiorgan bioassay for carcinogenesis with Wistar rats

A variety of chemicals can adversely affect the immune system and influence tumor development. The modifying potential of chemical carcinogens on the lymphoid organs and cytokine production of rats submitted to a medium-term initiation-promotion bioassay for carcinogenesis was investigated. Male Wistar rats were sequentially initiated with N-nitrosodiethylamine (DEN), N-methyl-N-nitrosourea (MNU), N-butyl-N-(4hydroxybutyl)nitrosamine (BBN), dihydroxy-di-n-propylnitrosamine (DHPN), and 1,2-dimethylhydrazine (DMH) during 4 weeks. Two initiated groups received phenobarbital (PB) or 2-acetyl amino fluorene (2-AAF) for 25 weeks and two noninitiated groups received only PB or 2-AAF. A nontreated group was used as control. Lymphohematopoietic organs, liver, kidneys, lung, intestines, and Zymbal's gland were removed for histological analysis. Interleukin (IL)-2, IL-12, interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), IL-10, and transforming growth factor betal (TGF-beta1) levels were determined by ELISA in spleen cell culture supernatants. At the fourth week, exposure to the initiating carcinogens resulted in cell depletion of the thymus, spleen and bone marrow, and impairment of IL-2, IL-12, and IFN-gamma production. However, at the 30th week, no important alterations were observed both in lymphoid organs and cytokine production in the different groups. The results indicate that the initiating carcinogens used in the present protocol exert toxic effects on the lymphoid organs and affect the production of cytokines at the initiation step of carcinogenesis. This early and reversible depression of the immune surveillance may contribute to the survival of initiated cells facilitating the development of future neoplasia. (C) 2003 Elsevier B.V. All rights reserved.

Lack of Epstein-Barr virus infection in cervical carcinomas

Context.-The Epstein-Barr virus (EBV) is a ubiquitous microorganism strongly associated with lymphoproliferative disorders and a large number of human neoplasms, mainly undifferentiated nasopharyngeal carcinoma and Burkitt lymphoma. The viral DNA has been detected in other tumors, such as carcinomas from tonsil, salivary glands, and thymus, and malignancies of the female genital tract. Some authors have proposed that EBV could play a role in the carcinogenesis of cervical tumors; however, other studies do not support this hypothesis.Objective.-To assess whether EBV is associated with female genital tract neoplasms.Design.-Sixty-five biopsy specimens (5 in situ carcinomas, 24 invasive squamous cell carcinomas, 6 lymphoepithelioma-like carcinomas, and 30 endocervical adenocarcinomas) were used to perform EBV detection through RNA in situ hybridization.Results.-None of the cervical carcinoma cases studied was positive for EBV infection.Conclusions.-The results suggest that it is still premature to incriminate EBV in the carcinogenesis of cervical carcinoma.

Detection of turkey astrovirus in young poults affected with poult enteritis complex in Brazil

Reverse transcription polymerase chain reaction (RT-PCR) of turkey astrovirus (TAstV) capsid and polymerase genes was applied to the bursa of Fabricius (BF), thymus (TH), spleen (SP) and cloacal swabs (CS) of young poults with "Poult enteritis complex" (PEC). The histological lesions included atrophy, lymphoid depletion, cellular infiltration and necrosis of the BF, TH and SP, respectively. The RT-PCR reactions were positive for the polymerase gene of TAstV-2 in all 100 CSs, 7 out of 10 of BFs and 10 out of 20 THs and SPs, respectively. Five out of 10 THs and SPs samples, considered to be negative by RT-PCR, were positive when specific primers designed for the TAstV-2 capsid gene were applied. This is the first description of turkey astrovirus infection presenting PEC in Latin America.

Direct detection of infectious bursal disease virus from clinical samples by in situ reverse transcriptase-linked polymerase chain reaction

The presence of the very virulent (vv) Brazilian strain of infectious bursal disease virus (IBDV) was determined in the bursa of Fabricius, thymus and liver of 2-week-old broilers from a flock with a higher than expected mortality. For this purpose, a direct in situ reverse transcriptase (RT)-linked polymerase chain reaction (PCR) method was developed using specific primers for vvIBDV. Unlabelled forward and reverse biotinylated oligonucleotides were used for RT-PCR in a one-step method and the respective products were revealed by a direct enzymatic reaction. The results were compared with those obtained by standard RT-PCR using general primers for IBDV and virus isolation. The virus isolation, RT-PCR and in situ RT-PCR revealed positive results on the bursa of Fabricius in 86%, 80% and 100%, respectively. The in situ RT-PCR detected vvIBDV in all tested thymus and liver samples, whereas the standard RT-PCR detected virus in 80% and 90% of the samples, respectively. After three consecutive passages on chicken embryonated eggs, IBDV was isolated from 64% of the thymus samples and 30% of the liver samples. In the present study, no classical or antigenic variants of IBDV were detected. The developed in situ RT-PCR assay was able to detect the very virulent strain of IBDV with a higher sensitivity than the conventional RT-PCR and virus isolation.

Atividade antifúngica de óleos essenciais frente a amostras clínicas de Candida albicans isoladas de pacientes HIV positivos

Objetivou-se avaliar a atividade antifúngica dos óleos essenciais de Ocimum basilicum L. (manjericão), Cymbopogon martinii L. (palmarosa), Thymus vulgaris L. (tomilho) e Cinnamomum cassia Blume (canela da china) sobre cepas de Candida albicans isoladas de pacientes HIV positivos e cepa padrão (ATCC 76845). Quinze amostras clínicas de C. albicans (C1-C15) foram repicadas em ágar Sabouraud Dextrose, para confecção de suspensões em solução salina estéril (0,9%) contendo 1,5 x 10(6) UFC mL-1. As emulsões dos óleos essenciais foram preparadas em água destilada estéril e tween 80, com concentrações variando entre 1024 µg mL-1 e 4 µg mL-1. A ação antifúngica foi determinada por meio da Concentração Inibitória Mínima (CIM) utilizando-se a técnica da microdiluição. Foram utilizados como controles positivos a nistatina e o miconazol (50 µg mL-1). Os testes foram realizados em triplicata, sendo a CIM, a menor concentração capaz de inibir o crescimento das leveduras, observada por método visual de acordo com a turvação do meio de cultura. Para C. albicans (ATCC 76845), a CIM do óleo essencial de C. cassia foi 64 µg mL-1, enquanto para óleo de C. martinii foi 1024 µg mL-1. Para as cepas clínicas, verificou-se que a CIM de C. cassia para 80% das cepas foi 64 µg mL-1, sendo a variação dos valores da CIM entre 128 µg mL-1 e 64 µg mL-1. Observou-se que para 66,6% das amostras clínicas, a CIM de C. martinii foi 612 µg mL-1. Constatou-se que a nistatina não apresentou atividade frente às cepas clínicas (C1-C15), enquanto a atividade antifúngica do miconazol foi verificada em 100% das amostras. Não se constatou atividade antimicrobiana dos óleos essenciais de O. basilicum e T. vulgaris, nas concentrações avaliadas. Concluiu-se que os óleos essenciais de C. cassia e C. martinii, em diferentes concentrações, apresentam atividade antifúngica sobre cepas de C. albicans isoladas de pacientes HIV positivos e cepa padrão (ATCC 76845). Entretanto não foi observada inibição antimicrobiana para os óleos de O. basilicum e T. vulgaris.