Avalia??o da resposta inflamat?ria cardiovascular em ratos com hipertens?o renovascular (2R1C) infectados pelo Trypanosoma cruzi.

Programa de P?s-Gradua??o em Ci?ncias Biol?gicas. N?cleo de Pesquisas em Ci?ncias Biol?gicas, Pr?-Reitoria de Pesquisa de P?s Gradua??o, Universidade Federal de Ouro Preto.A Hipertens?o Arterial Sist?mica (HAS), caracterizada pelo aumento de Angiotensina II plasm?tica/tecidual, ? uma das doen?as com maior preval?ncia no mundo moderno. Evidencias apontam para o envolvimento da inflama??o vascular no processo inicial da HAS. A cardiopatia chag?sica ? causada pelo protozo?rio Trypanosoma cruzi, capaz de desencadear uma resposta inflamat?ria progressiva gerando importantes altera??es el?tricas e morfo-funcionais ao cora??o. Nosso objetivo neste estudo foi avaliar a associa??o da HAS e os aspectos inflamat?rios cardiovasculares em animais infectados pela cepa Y do T. cruzi. Ratos Wistars machos foram submetidos ? cirurgia renovascular (modelo Goldblatt ? 2Rins/1CLIP) ou ? cirurgia fict?cia (SHAM) e divididos e agrupados de acordo com a infec??o ou n?o pelo T. cruzi. Foram realizados tr?s experimentos em diferentes momentos da ?infec??o e cirurgia? para avalia??o da HAS e do patol?gico induzido pelo parasito sendo (I) indu??o da HAS + infec??o pelo T.cruzi (ao mesmo tempo), experimento teve dura??o de 1 semana; (II) indu??o da HAS + infec??o com o T. cruzi 3 semanas ap?s a cirurgia e (III) indu??o da HAS + infec??o pelo T.cruzi (ao mesmo tempo) e experimento teve dura??o de 8 semanas. Nesse estudo, avaliou-se par?metros fisiol?gicos e inflamat?rios como pletismografia, Ang II e mediadores inflamat?rios plasm?ticos, parasitemia e achados histopatol?gicos no tecido card?aco. Ap?s a 3? semana de cirurgia os animais apresentaram picos press?ricos havendo concord?ncia com a produ??o de Angio II plasm?tica nas primeiras semanas, mas n?o se observando rela??o direta entre o aumento da press?o arterial e o numero de parasitos circulantes. No entanto, o T. cruzi exerceu papel definidor na alta produ??o de TNF-alfa e da CX3CL1/Fractalkine e, em associa??o com a cirurgia renovascular, observou-se maior eleva??o nesses mediadores inflamat?rios em diferentes etapas do estudo. Por outro lado, o aumento da press?o arterial n?o exerceu interfer?ncia no processo inflamat?rio tecidual nem na invas?o tecidual pelo parasito, mas contribuiu para a manuten??o do n?mero de vasos sangu?neos ap?s 8 semanas de experimento, quando houve diminui??o no grupo de animais normotensos. Com este estudo conclui-se que o tempo de infec??o pelo T. cruzi e o tempo de desenvolvimento da hipertens?o interferem, mutualmente, no perfil inflamat?rio induzido pelo hospedeiro mam?fero. No entanto, acredita-se tamb?m, que esse estudo fosse reproduzido em outro modelo experimental mais suscept?vel ? infec??o pelo T. cruzi, as les?es cardiovasculares seriam mais evidenciadas principalmente numa etapa cr?nica de ambas as doen?as.Systemic arterial hypertension (SAH) is characterized by the increase of plasma/tissue Angiotensin II (Angio II) and presents high prevalence in the world. There are evidences suggesting the involvement of vascular inflammation in the initial phase of SAH. Chagas cardiomyopathy is a disease caused by Trypanosoma cruzi infection that triggers a progressive inflammatory response with prominent fibrosis and, as a consequence, there are important cardiac electrical and functional disturbances and/or hypertrophy in this organ. Our aim in this study was evaluate SAH and the cardiovascular inflammation in Wistar rats infected by Y strain of T. cruzi. Male rats were submitted to renovascular surgery (Goldblat model ? 2Kidneys/1CLIP) or to a fictional surgery (SHAM) and grouped according to infection or not by T. cruzi. It was proposed 3 experiments in different times of infection and surgery to evaluate the SAH and the pathological aspects induzed by the parasite: (I) SAH induction and T.cruzi infection at same time ? experiment occurred in one week; (II) SAH induction + T.cruzi infection after 3 weeks of surgery and (III) SAH induction and T.cruzi infection at same time ? experiment occurred during 8 weeks. For these sets of experiments, there was evaluation of physiological, parasitological, inflammatory and histological parameters in the plasma and/or cardiac tissue. After the 3rd week of the renovascular surgery, animals presented high pressoric levels in accordance with high plasma Angio II on the first weeks. However, there were not observed relation between the increase of SAH and the number of circulating parasites. In our data, T. cruzi exerted an important role in the production of TNF-alpha and CX3CL1/Fractalkine by host cells and we observed high plasma release of these mediators when associated with SAH, in distinct times of this study. On the other hand, SAH does not exerted influence on cardiac inflammatory infiltration neither in the amastigote proliferation, but contributed to the maintenance of the number of blood vessels after 8 weeks of the experiment in comparison with the reduction observed in the control group (without SAH). Together, these data suggest that the time of T.cruzi infection and the time of SAH development interfere, mutually, in the inflammatory profile induced by host mammalian. However, other studies performed in other T.cruzi-susceptible experimental model might develop cardiovascular lesions with high magnificence as well as inflammatory process in heart tissue during chronic stages of both diseases

A novel microtubule de-stabilizing complementarity-determining region C36L1 peptide displays antitumor activity against melanoma in vitro and in vivo

Short peptide sequences from complementarity-determining regions (CDRs) of different immunoglobulins may exert anti-infective, immunomodulatory and antitumor activities regardless of the specificity of the original monoclonal antibody (mAb). In this sense, they resemble early molecules of innate immunity. C36L1 was identified as a bioactive light-chain CDR1 peptide by screening 19 conserved CDR sequences targeting murine B16F10-Nex2 melanoma. The 17-amino acid peptide is readily taken up by melanoma cells and acts on microtubules causing depolymerization, stress of the endoplasmic reticulum and intrinsic apoptosis. At low concentrations, C36L1 inhibited migration, invasion and proliferation of B16F10-Nex2 cells with cell cycle arrest at G2/M phase, by regulating the PI3K/Akt signaling axis involving Rho-GTPase and PTEN mediation. Peritumor injection of the peptide delayed growth of subcutaneously grafted melanoma cells. Intraperitoneal administration of C36L1 induced a significant immune-response dependent anti-tumor protection in a syngeneic metastatic melanoma model. Dendritic cells stimulated ex-vivo by the peptide and transferred to animals challenged with tumor cells were equally effective. The C36 VL CDR1 peptide is a promising microtubule-interacting drug that induces tumor cell death by apoptosis and inhibits metastases of highly aggressive melanoma cells

Diversidade e variação sazonal de moscas-das-frutas (Diptera: Tephritidae, Lonchaeidae) e seus parasitóides (Hymenoptera: Braconidae, Figitidae) em pomares de goiaba, nêspera e pêssego

This work was carried out in orchards of guava progenies, and loquat and peach cultivars, in Monte Alegre do Sul, SP, Brazil, in 2002 and 2003. Guavas and loquats were bagged and unbagged bi-weekly and weekly, respectively, for assessment of the infestation period. Peach was only bagged weekly. The assays started when the fruits were at the beginning of development, but still green. Ripe fruits were taken to the laboratory and placed individually into plastic cups. McPhail plastic traps containing torula yeast were hung from January 2002 to January 2004 to assess the fruit fly population in each orchard, but only the Ceratitis capitata population is here discussed. Five tephritid species were reared from the fruits: Anastrepha bistrigata Bezzi, A. fraterculus (Wiedemann), A. obliqua (Macquart), A. sororcula Zucchi, and C. capitata, in addition to six lonchaeid species: Neosilba certa (Walker), N. glaberrima (Wiedemann), N. pendula (Bezzi), N. zadolicha McAlpine and Steyskal, Neosilba sp. 4, and Neosilba sp. 10 (both species are in the process of being described by P. C. Strikis), as well as some unidentified Neosilba species. Ten parasitoid species were obtained from fruit fly puparia, of which five were braconids: Asobara anastrephae (Muesebeck), Doryctobracon areolatus (Szépligeti), D. brasiliensis (Szépligeti), Opius bellus Gahan, and Utetes anastrephae (Viereck), and five figitids: Aganaspis pelleranoi (Brèthes), Dicerataspis grenadensis Ashmead, Lopheucoila anastrephae (Rhower), Leptopilina boulardi (Barbotin, Carlton and Kelner-Pillaut), and Trybliographa infuscata Diaz, Gallardo and Uchôa. Ceratitis capitata showed a seasonal behavior with population density peaking at the second semester of each year. Anastrepha and Neosilba species remained in the orchards throughout both years.Este trabalho foi realizado em três pomares em Monte Alegre do Sul, SP, em 2002 e 2003, representados por coleção de progênies de goiabeiras, de cultivares de nespereiras e de cultivares de pessegueiros. O período de infestação foi determinado por meio de ensacamento e desensacamento quinzenal e semanal de goiabas e nêsperas, respectivamente, e pelo ensacamento semanal de pêssegos. Os ensaios iniciaram-se com os frutos verdes (princípio de desenvolvimento). Os frutos maduros foram levados ao laboratório e acondicionados individualmente em copos plásticos. A flutuação populacional de Ceratitis capitata (Wiedemann) foi avaliada por meio de armadilhas plásticas modelo McPhail com torula em cada pomar, de janeiro/2002 a janeiro/2004. Dos frutos foram obtidas cinco espécies de tefritídeos: Anastrepha bistrigata Bezzi, A. fraterculus (Wiedemann), A. obliqua (Macquart), A. sororcula Zucchi e C. capitata e seis de lonqueídeos: Neosilba certa (Walker), N. glaberrima (Wiedemann), N. pendula (Bezzi), N. zadolicha McAlpine and Steyskal, Neosilba sp. 4 e Neosilba sp. 10, além de algumas espécies não-identificadas. Foram obtidas 10 espécies de parasitóides, cinco da família Braconidae - Asobara anastrephae (Muesebeck), Doryctobracon areolatus (Szépligeti), D. brasiliensis (Szépligeti), Opius bellus Gahan e Utetes anastrephae (Viereck) - e cinco da família Figitidae - Aganaspis pelleranoi (Brèthes), Dicerataspis grenadensis Ashmead, Lopheucoila anastrephae (Rhower), Leptopilina boulardi (Barbotin, Carlton and Kelner-Pillaut) e Trybliographa infuscata Diaz, Gallardo and Uchôa. Ceratitis capitata apresentou comportamento sazonal com picos populacionais durante o segundo semestre dos dois anos. As espécies de Anastrepha e de Neosilba permaneceram nos pomares durante os dois anos

Production of monoclonal antibodies for detection of a secreted aspartyl proteinase from Candida spp. in biologic specimens

Secreted acid proteinases (SAP) constitute an important group of virulence factors in Candida albicans. In the present work, an acid proteinase from C. albicans was sequentially purified from the supernatant of a yeast culture by precipitation with ammonium sulfate, ion exchange chromatography, and molecular exclusion chromatography, yielding a specific enzymatic activity of 204.1 IU/mg on bovine serum albumin (BSA). The molecular mass of the purified proteinase was estimated at 43 kd after exclusion chromatography and at 41 kd by nondenaturating sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The purified proteinase was able to degrade BSA at pH 2.5, but was not active on collagen, and it was significantly inhibited by pepstatin A. The immunization of BALB/c mice with the purified proteinase and later fusion of their spleen cells with myeloma cells resulted in 19 monoclonal antibody secreting hybridomas (MAbs) capable of detecting SAP in enzyme-linked immunosorbent assay ( ELISA) assays. All MAbs obtained are isotype IgG1 kappa (kappa) immunoglobulins and develop a 41 kd protein band by Western blot (WB) in samples of SAP obtained from C. albicans (12-A) and C. dubliniensis ( strain 778) crude extracts. The anti-SAP MAbs were used in capture ELISA and two combinations of these antibodies proved suitable for SAP detection, that is, MAP1 (1B1B3) or MAP2 (2D2C10) as coat antibodies, and biotinylated MAP3 (2A6E8) as detect antibody. Capture ELISA using these sets of MAbs detected over 32 ng/mL protein in purified SAP samples as well as in crude C. albicans and C. dubliniensis extracts. The results herein obtained allow for the prediction of how this set of antibodies can be useful for SAP detection in biologic specimens.26420120

Blockade of MIF-CD74 Signalling on Macrophages and Dendritic Cells Restores the Antitumour Immune Response Against Metastatic Melanoma

Mounting an effective immune response against cancer requires the activation of innate and adaptive immune cells. Metastatic melanoma is the most aggressive form of skin cancer. While immunotherapies have shown a remarkable success in melanoma treatment, patients develop resistance by mechanisms that include the establishment of an immune suppressive tumor microenvironment. Thus, understanding how metastatic melanoma cells suppress the immune system is vital to develop effective immunotherapies against this disease. In this study, we find that macrophages (MOs) and dendritic cells (DCs) are suppressed in metastatic melanoma and that the Ig-CDR-based peptide C36L1 is able to restore MOs and DCs' antitumorigenic and immunogenic functions and to inhibit metastatic growth in lungs. Specifically, C36L1 treatment is able to repolarize M2-like immunosuppressive MOs into M1-like antitumorigenic MOs, and increase the number of immunogenic DCs, and activated cytotoxic T cells, while reducing the number of regulatory T cells and monocytic myeloid-derived suppressor cells in metastatic lungs. Mechanistically, we find that C36L1 directly binds to the MIF receptor CD74 which is expressed on MOs and DCs, disturbing CD74 structural dynamics and inhibiting MIF signaling on these cells. Interfering with MIF-CD74 signaling on MOs and DCs leads to a decrease in the expression of immunosuppressive factors from MOs and an increase in the capacity of DCs to activate cytotoxic T cells. Our findings suggest that interfering with MIF-CD74 immunosuppressive signaling in MOs and DCs, using peptide-based immunotherapy can restore the antitumor immune response in metastatic melanoma. Our study provides the rationale for further development of peptide-based therapies to restore the antitumor immune response in metastatic melanoma

In vitro bioactivity of titanium-doped bioglass

Previous studies have suggested that incorporating relatively small quantities of titanium dioxide into bioactive glasses may result in an increase in bioactivity and hydroxyapatite formation. The present work therefore investigated the in vitro bioactivity of a titanium doped bioglass and compared the results with 45S5 bioglass. Apatite formation was evaluated for bioglass and Ti-bioglass in the presence and absence of foetal calf serum. Scanning electron microscopy (SEM) images were used to evaluate the surface development and energy dispersive X-ray measurements provided information on the elemental ratios. X-ray diffraction spectra confirmed the presence of apatite formation. Cell viability was assessed for bone marrow stromal cells under direct and indirect contact conditions and cell adhesion was assessed using SEM