PATRICK JACK SPENCER

Resumo

Possui graduação em Ciências Biológicas pela Universidade Presbiteriana Mackenzie (1991), mestrado em Tecnologia Nuclear pela Universidade de São Paulo (1995) e doutorado em Tecnologia Nuclear pela Universidade de São Paulo (2000) tendo sido bolsista sandwich no US Army Medical Research Institute for Infeccious Diseases (98-99). É responsável pelo Biotério de criação e manutenção de animais de laboratório do IPEN. Tem experiência na área de Bioquímica, com ênfase em Proteínas, atuando principalmente nos seguintes temas: veneno, proteínas, bothrops, irradiação e miotoxina.(Texto extraído do Currículo Lattes em 22 dez. 2021)

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Agora exibindo 1 - 8 de 8
  • Artigo IPEN-doc 29948
    Comparing traditional and toxin-oriented approaches towards antivenom production against Bitis arietans snake venom
    2023 - GUIDOLIN, FELIPE R.; GODOI, KEMILY S. de; MEGALE, ANGELA A.A.; SILVA, CRISTIANE C.F. da; KODAMA, ROBERTO T.; CAJADO-CARVALHO, DANIELA; IWAI, LEO K.; SPENCER, PATRICK J.; PORTARO, FERNANDA C.V.; SILVA, WILMAR D. da
    Accidents with snakes are responsible for about 32,000 deaths annually in sub-Saharan Africa, caused mostly by snakes from the genus Bitis, in particular Bitis arietans. B. arietans venom is composed of a complex mixture of toxins, mainly metalloproteases, serine proteases, phospholipases, lectins, and disintegrins. In this work, we compared two approaches to anti-B. arietans antivenom production: immunization with crude snake venom (“traditional approach”) and immunization with selected key toxins isolated from the snake venom (“toxin oriented” approach). Fractions from B. arietans venom were isolated by size exclusion chromatography. Crude venom and samples containing serine proteases or metalloproteases were selected for the immunization of BALB/c mice. Anti-B. arietans and anti-serine proteases plasmas showed a similar recognition profile and higher titers and affinity than the anti-metalloproteases plasma. Cross-recognition of other Bitis venoms was observed, but with low intensity. Although the plasma of all experimental groups inhibited the enzymatic activity of B. arietans venom in vitro, in vivo protection was not achieved. Our results have shown limitations in both approaches considered. Based on this, we proposed a model of polyclonal, species-specific, monovalent antivenoms that could be used as a base to produce customizable polyvalent sera for use in sub-Saharan Africa.
  • Artigo IPEN-doc 29689
    Anti-Metalloproteases
    2023 - GODOI, KEMILY S. de; GUIDOLIN, FELIPE R.; PORTARO, FERNANDA C.V.; SPENCER, PATRICK J.; SILVA, WILMAR D. da
    Bitis arietans is a medically important snake found in Sub-Saharan Africa. The envenomation is characterized by local and systemic effects, and the lack of antivenoms aggravates the treatment. This study aimed to identify venom toxins and develop antitoxins. The F2 fraction obtained from Bitis arietans venom (BaV) demonstrated the presence of several proteins in its composition, including metalloproteases. Titration assays carried out together with the immunization of mice demonstrated the development of anti-F2 fraction antibodies by the animals. The determination of the affinity of antibodies against different Bitis venoms was evaluated, revealing that only BaV had peptides recognized by anti-F2 fraction antibodies. In vivo analyses demonstrated the hemorrhagic capacity of the venom and the effectiveness of the antibodies in inhibiting up to 80% of the hemorrhage and 0% of the lethality caused by BaV. Together, the data indicate: (1) the prevalence of proteins that influence hemostasis and envenomation; (2) the effectiveness of antibodies in inhibiting specific activities of BaV; and (3) isolation and characterization of toxins can become crucial steps in the development of new alternative treatments. Thus, the results obtained help in understand
  • Artigo IPEN-doc 29112
    Antibodies as snakebite antivenoms
    2022 - SILVA, WILMAR D. da; ANDRADE, SONIA A. de; MEGALE, ANGELA A.A.; SOUZA, DANIEL A. de; SANTANNA, OSVALDO A.; MAGNOLI, FABIO C.; GUIDOLIN, FELIPE R.; GODOI, KEMILY S.; SALADINI, LUCAS Y.; SPENCER, PATRICK J.; PORTARO, FERNANDA C.V.
    Snakebite envenomation is considered a neglected tropical disease, affecting tens of thousands of people each year. The recommended treatment is the use of antivenom, which is composed of immunoglobulins or immunoglobulin fragments obtained from the plasma of animals hyperimmunized with one (monospecific) or several (polyspecific) venoms. In this review, the efforts made in the improvement of the already available antivenoms and the development of new antivenoms, focusing on snakes of medical importance from sub-Saharan Africa and Latin America, are described. Some antivenoms currently used are composed of whole IgGs, whereas others use F(ab’)2 fragments. The classic methods of attaining snake antivenoms are presented, in addition to new strategies to improve their effectiveness. Punctual changes in immunization protocols, in addition to the use of cross-reactivity between venoms from different snakes for the manufacture of more potent and widely used antivenoms, are presented. It is known that venoms are a complex mixture of components; however, advances in the field of antivenoms have shown that there are key toxins that, if effectively blocked, are capable of reversing the condition of in vivo envenomation. These studies provide an opportunity for the use of monoclonal antibodies in the development of new-generation antivenoms. Thus, monoclonal antibodies and their fragments are described as a possible alternative for the production of antivenoms, regardless of the venom. This review also highlights the challenges associated with their development.
  • Artigo IPEN-doc 28693
    Synthesis, in vitro testing, and biodistribution of surfactant-free radioactive nanoparticles for cancer treatment
    2022 - SOUZA, CARLA D. de; BARBEZAN, ANGELICA B.; ROSERO, WILMMER A.A.; SANTOS, SOFIA N. dos; CARVALHO, DIEGO V. de S.; ZEITUNI, CARLOS A.; BERNARDES, EMERSON S.; VIEIRA, DANIEL P.; SPENCER, PATRICK J.; RIBEIRO, MARTHA S.; ROSTELATO, MARIA E.C.M.
    New forms of cancer treatment, which are effective, have simple manufacturing processes, and easily transportable, are of the utmost necessity. In this work, a methodology for the synthesis of radioactive Gold-198 nanoparticles without the use of surfactants was described. The nuclear activated Gold-198 foils were transformed into H198AuCl4 by dissolution using aqua regia, following a set of steps in a specially designed leak-tight setup. Gold-198 nanoparticles were synthesized using a citrate reduction stabilized with PEG. In addition, TEM results for the non-radioactive product presented an average size of 11.0 nm. The DLS and results for the radioactive 198AuNPs presented an average size of 8.7 nm. Moreover, the DLS results for the PEG-198AuNPs presented a 32.6 nm average size. Cell line tests showed no cytotoxic effect in any period and the concentrations were evaluated. Furthermore, in vivo testing showed a high biological uptake in the tumor and a cancer growth arrest.
  • Artigo IPEN-doc 26495
    Proteomic analysis of soluble proteins retrieved from Duttaphrynus melanostictus skin secretion by IEx-batch sample preparation
    2019 - MARIANO, DOUGLAS O.C.; PREZOTTO-NETO, JOSE P.; SPENCER, PATRICK J.; SCIANI, JULIANA M.; PIMENTA, DANIEL C.
    Amphibians display a toxic secretion that works as chemical defenses against predators and/or microorganisms that is stored in specialized glands located in the tegument. For some animals, such glands have accumulated in specific regions of the body and formed prominent structures known as macroglands. The Bufonidae family displays conspicuous macroglands in a post-orbital position, termed parotoids, which secretions are known to be extremely viscous and rich in alkaloids and steroids. Few proteins have been described in this material, though. Mainly, because of the difficulties to handle such biological matrix. In this context, we have performed a proteomic study on the parotoid macrogland secretion of the Asian bufonid Duttaphrynus melanostictus. By employing the Ion-Exchange (IEx)-batch chromatography (anionic, cationic and both) we obtained six fractions - bound and unbound – that were submitted to an in solution-trypsin digestion followed by LC-MS/MS. Proteins related to: antioxidant activity, binding processes (carbohydrate/lipid/protein), energy metabolism, hydrolases, lipid metabolism and membrane traffic were identified. Moreover, IEx was able to preserve the biological activity of the retrieved proteins (peptidasic). The current study increases the knowledge on the proteins present in the bufonids parotoid macrogland secretion, providing a better understanding of the physiological role played by such molecules. Significance: The current approach allowed a detailed proteomic analysis of the several proteins synthesized in the D. melanostictus parotoid macrogland (Bufonidae) that are secreted into the skins, but embedded within a complex viscous biological matrix. Moreover, our results aim to increase the knowledge on the biological role played by such proteins at the skin.
  • Artigo IPEN-doc 23048
    Protein profile analysis of two australian snake venoms by one- dimensional gel electrophoresis and MS/MS experiments
    2017 - GEORGIEVA, DESSISLAVA; HILDEBRAND, DIANA; SIMAS, RODRIGO; CORONADO, MONIKA A.; KWIATKOWSKI, MARCEL; SCHLUTER, HARTMUT; ARNI, RAGHUVIR; SPENCER, PATRICK; BETZEL, CHRISTIAN
    The Pseudechis colletti and Pseudechis butleri venoms were analyzed by 1-D gel electrophoresis, followed by mass spectrometric analysis of tryptic peptides obtained from the protein bands. Both venoms contain highly potent pharmacologically active components, which were assigned to the following protein families: basic and acidic phospholipases A2 (PLA2s), L-amino acid oxidases (LAAOs), P-III metalloproteinases (P-III SVMPs), 5’- nucleotidases (5’-NTDs), cysteine-rich secretory proteins (CRISPs), venom nerve growth factors (VNGFs) and post-synaptic neurotoxins. Considerable predominance of PLA2s over other toxins is a characteristic feature of both venoms. The major differences in the venom compositions are the higher concentration of SVMPs and CRISPs in the P. butleri venom, as well as the presence of post-synaptic neurotoxins. Furthermore, the analysis revealed a high concentration of proteins with myotoxic, coagulopathic and apoptotic activities. PLA2s are responsible for the myotoxic and anticoagulant effects observed in patients after envenomation (4). The other protein families, encountered in the two venoms, probably contribute to the major symptoms described for these venoms. These results explain the observed clinical effects of the black snake envenomation. The analyzed venoms contain group P-III metalloproteinases of medical importance with the potency to be used for diagnostic purposes of von Willebrand factor (vWF) disease, for regulation of vWF in thrombosis and haemostasis, for studying the function of the complement system in host defense and in the pathogenesis of diseases. Comparison of venomic data showed similarities in the major venom components of snakes from the genus Pseudechis, resulting in common clinical effects of envenomation, and demonstrating close relationships between venom toxins of Elapidae snakes.
  • Artigo IPEN-doc 21327
    Venomics of the Australian eastern brown snake (Pseudonaja textilis): Detection of new venom proteins and splicing variants
    2015 - VIALA, VINCENT L.; HILDEBRAND, DIANA; TRUSCH, MARIA; FUCASE, TAMARA M.; SCIANI, JULIANA M.; PIMENTA, DANIEL C.; ARNI, RAGHUVIR K.; SCHLUTER, HARTMUT; BETZEL, CHRISTIAN; MIRTSCHIM, PETER; DUNSTAN, NATHAN; SPENCER, PATRICK J.
  • Artigo IPEN-doc 21288
    Proteomic analysis of the rare Uracoan rattlesnake Crotalus vegrandis venom: Evidence of a broad arsenal of toxins
    2015 - VIALA, VINCENT L.; HILDEBRAND, DIANA; FUCASE, TAMARA M.; SCIANI, JULIANA M.; PREZOTTO NETO, JOSE P.; RIEDNER, MARIA; SANCHES, LEONARDO; NISHIMURA, PAULA J.; OGUIURA, NANCY; PIMENTA, DANIEL C.; SCHLUTER, HARTMUT; BETZEL, CHRISTIAN; ARNI, RAGHUVIR K.; SPENCER, PATRICK J.