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REDE GOIANA DE PESQUISA EM TUBERCULOSE

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Programa de Pós Graduação em Medicina Tropical e Saúde Pública

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I Reunião Vacinas e biomarcadores INCT-TB/ REDE TB Hospital Clementino Fraga Filho -UFRJ
Por Ana Paula Junqueira Kipnis Em 18/09/2017 às 14:55

Objetivando definir estratégias para o melhor controle da TB o grupo responsável por vacinas e biomarcadores do INCT-TB juntamente com os coordenadores e pesquisadores dessas área da Rede TB se reunirão dia 20 de Setembro de 2017 no Rio de Janeiro.

missão de trabalho india
Colaboração Biotecnológica com a INDIA- CSRI-CDRI-
Por Ana Paula Junqueira Kipnis Em 11/08/2017 às 11:03

Entre aqui para ver os objetivos do projeto e as fotos da visita técnica a India.

DPOC
Artigos 2017: Do COPD and Healthy Subjects have Similar Acute Inflammatory Response Induced by Sub-maximum Effort Test?
Por Lázaro Moreira Marques Neto Em 04/07/2017 às 14:01

Autores: Krislainy De Sousa Corrêa , Adeliane Castro Da Costa , Ana Paula Perillo Ferreira Carvalho , José Laerte Rodrigues Da Silva Júnior , Maria Rosedália De Moraes , Ana Paula Junqueira-Kipnis and Marcelo Fouad Rabahi

Abstract

Introduction: COPD (chronic obstructive pulmonary disease) presents a low degree of systemic inflammation responsible for the disease’s extra pulmonary consequences. Its inflammatory profile can be altered by acute exercise. However, acute effects of a sub maximum test, capable of reproducing activities of daily living (ADL) in sedentary COPD individuals, are not well-known

Objective: To evaluate if the six-minute walk test (6MWT) is capable of altering interleukin-6 (IL-6) blood levels, tumoral necrosis factor alfa (TNF-α) and hypersensitive C reactive protein (hs-CRP) in relation to the basal level of COPD individuals.

Methods: 21 individuals with moderate and severe COPD and 8 healthy individuals, with no history of smoking, sedentary, matched by age were assessed regarding plasma levels of IL-6, TNF-α and hs-CRP before and right after a 6MWT. They also did spirometry and body composition analysis.

Results: 6MWT did not provoke significant alteration in IL-6 levels in COPD individuals (pre=4.53 ± 9.0 pg/ml vs. post=7.14 ± 11.31 pg/ml, p=0.11), whereas in healthy individuals this increase was significant (pre=1.56 ± 6.45 pg/ml vs. post=4.37 ± 8.0 pg/ml, p<0.01). COPD individuals present higher hs-CRP blood levels when compared to healthy subjects both in rest (8.15 ± 9.68 pg/ml vs. 2.60 ± 1.88 pg/ml, p=0.02), and after exercise (8.18 ± 9.08 pg/ml vs. 2.62 ± 1.85 pg/ml, p=0.02. TNF-α did not present difference between groups during rest (COPD=2.13 ± 1.03 pg/ml vs. healthy=2.00 ± 0.59 pg/ml, p>0.05) as well as after 6MWT (COPD=2.48 ± 1.92 pg/ml vs. healthy=1.89 ± 0.69 pg/ml, p>0.05), and their intragroup focus was not affected by the effort (p>0.05).

Conclusions: In COPD patients, 6MWT does not induce acute inflammatory response of IL-6 at the same proportion of that of healthy subjects. Hs-CRP’s and TNFα’s response to 6MWT was similar between groups. COPD patients presented higher concentrations of hs-CRP than healthy individuals.

 

O texto completo pode ser encontrado no link:

https://www.omicsonline.org/open-access/do-copd-and-healthy-subjects-have-similar-acute-inflammatory-responseinduced-by-submaximum-effort-test.php?aid=87058

Eduardo CMX
Artigos 2017: Identification of specific antibodies against the Ag85C-MPT51-HspX fusion protein (CMX) for serological screening of tuberculosis in endemic area.
Por Lázaro Moreira Marques Neto Em 04/07/2017 às 08:45

Autores: Zagmignan ACosta ACDViana JLLima Neto LGMonteiro CAGaioso Neto AGJunqueira-Kipnis APde Sousa EM.

Abstract

OBJECTIVES:

Development of new tools for rapid and accurate diagnosis of tuberculosis (TB) is considered a strategy for controlling the disease. The recombinant CMX fusion protein is composed of immunodominant epitopes of the Ag85C (Rv0129c), MPT51 (Rv3803c) and the entire HspX (Rv2031c) proteins from Mycobacterium tuberculosis H37Rv (Mtb). The aim of this study was to evaluate the applicability of a test using the CMX protein in individuals suspected of TB.

METHODS:

Indirect ELISA was used to measure serum anti-CMX IgM and IgG in individuals with pulmonary TB.

RESULTS:

Patients with pulmonary TB had higher titers of IgM (OD = 0.502 ± 0.281) than healthy controls (OD = 0.200 ± 0.125). The cutoff for IgM-ELISA was determined using ROC curve analyzes (AUC = 0.868) with a sensitivity of 80.1% and a specificity of 78.2%. Patients with pulmonary TB also had higher titers of IgG (OD = 0.525 ± 0.391) than healthy controls (OD = 0.215 ± 0.077). The cutoff for IgG-ELISA was determined using ROC curve analyzes (AUC = 0.864) with a sensitivity of 81.7% and a specificity of 74.7%.

CONCLUSION:

The results suggest that the recombinant protein CMX can be used in a serological test to complement the screening of individuals suspected of having active pulmonary TB.

 

O artigo completo pode ser encontrado no link:

http://www.tandfonline.com/doi/abs/10.1080/1744666X.2017.1345626?journalCode=ierm20

BCG - Luciana Leite
Artigos 2017: Recombinant BCG Expressing LTAK63 Adjuvant induces Superior Protection against Mycobacterium tuberculosis.
Por Lázaro Moreira Marques Neto Em 04/07/2017 às 08:35

Autores: Nascimento IPRodriguez DSantos CCAmaral EPRofatto HKJunqueira-Kipnis APGonçalves EDCD'Império-Lima MRHirata MHSilva CLWinter NGicquel BMills KHGPizza MRappuoli RLeite LCC.

In order to develop an improved BCG vaccine against tuberculosis we have taken advantage of the adjuvant properties of a non-toxic derivative of Escherichia coli heat labile enterotoxin (LT), LTAK63. We have constructed rBCG strains expressing LTAK63 at different expression levels. Mice immunized with BCG expressing low levels of LTAK63 (rBCG-LTAK63lo) showed higher Th1 cytokines and IL-17 in the lungs, and when challenged intratracheally with Mycobacterium tuberculosis displayed a 2.0-3.0 log reduction in CFU as compared to wild type BCG. Histopathological analysis of lung tissues from protected mice revealed a reduced inflammatory response. Immunization with rBCG-LTAK63lo also protected against a 100-fold higher challenge dose. Mice immunized with rBCG-LTAK63lo produced an increase in TGF-β as compared with BCG after challenge, with a corresponding reduction in Th1 and Th17 cytokines, as determined by Real Time RT-PCR. Furthermore, rBCG-LTAK63lo also displays protection against challenge with a highly virulent Beijing isolate. Our findings suggest that BCG with low-level expression of the LTAK63 adjuvant induces a stronger immune response in the lungs conferring higher levels of protection, and a novel mechanism subsequently triggers a regulatory immune response, which then limits the pathology. The rBCG-LTAK63lo strain can be the basis of an improved vaccine against tuberculosis.

 

O artigo completo pode ser acessado pelo link:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5437048/

balmani
Artigos 2017: Acinetobacter baumannii strains isolated from patients in intensive care units in Goiânia, Brazil: Molecular and drug susceptibility profiles.
Por Lázaro Moreira Marques Neto Em 19/05/2017 às 11:30

Abstract

Resistance to antimicrobial agents is increasing worldwide and imposes significant life-threatening risks to several different populations, especially those in intensive care units (ICUs). Bacteria can quickly develop or acquire resistance to antimicrobial drugs, and combined with their intrinsic potential to cause disease in humans, these bacteria can become deadly. Among Gram-negative bacteria, Acinetobacter baumannii is notorious as a frequent opportunistic pathogen associated with critically ill patients, and understanding the genetic basis of A. baumannii resistance to beta-lactams among patients in ICUs will result in better protocols to prevent the development of resistance as well as improved treatment regimens. In this study, we assessed 1333 patients in five ICUs, 56 of whom developed A. baumannii infections. Most of the A. baumannii isolates were resistant to beta-lactam antimicrobial drugs, specifically, 3rd- and 4th-generation cephalosporins and carbapenems, and 91.1% of the isolates were multi-drug resistant (MDR). The most frequent OXA gene present was OXA-23 (55.1%), which is significantly associated with MDR strains. Most of the A. baumannii isolates (76.8%) were capable of forming a biofilm. The antimicrobial drug classes that were effective against most of these isolates were polymyxins and tigecycline. The molecular profile of the isolates allowed detection of 12 different clusters comprising 2 to 8 isolates each. In conclusion, our data indicate a high incidence of resistance to carbapenems as well as MDR strains among the observed A. baumannii isolates, most of which exhibited a high prevalence of OXA-23 gene expression. Only a few selective drugs were effective, reinforcing the notion that bacterial resistance is an emerging problem that should be prioritized in every healthcare facility.

 

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0176790

m0 modulation
Artigos 2017: Modulation of Macrophage Responses by CMX, a Fusion Protein Composed of Ag85c, MPT51, and HspX from Mycobacterium tuberculosis
Por Lázaro Moreira Marques Neto Em 19/05/2017 às 11:29

Abstract

Mycobacterium bovis Bacillus Calmette–Guérin (BCG) is a vaccine used to prevent tuberculosis (TB). Due to the poor protection conferred by BCG in adults, new, more effective formulations have been developed. A recombinant BCG vaccine expressing the CMX fusion protein Ag85c_MPT51_HspX (rBCG-CMX) induced Th1 and Th17 responses and provided better protection than BCG. It has been shown that Mycobacterium smegmatis expressing CMX also induces better protection than BCG and is a strong macrophage activator. The aim of the present study was to evaluate macrophage activation by the recombinant CMX fusion protein and by rBCG-CMX and to evaluate their ability to generate vaccine-specific immune responses. The results demonstrate that rCMX protein expressed by BCG (rBCG-CMX) activates pulmonary macrophages; increases the expression of activation molecules, cytokines, and MHC-II. The interaction with rCMX activates the transcription factor NF-κB and induces the production of the cytokines TGF-β, TNF-α, and IL-6. The in vitro stimulation of bone marrow-derived macrophages (BMMs) from TLR-4 or TLR-2 KO mice showed that in the absence of TLR-4, IL-6 was not produced. rBCG-CMX was unable to induce CMX-specific Th1 and Th17 cells in TLR-4 and TLR-2 KO mice, suggesting that these receptors participate in their induction. We concluded that both the rBCG-CMX vaccine and the rCMX protein can activate macrophages and favor the specific immune response necessary for this vaccine.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389097/

nano
Artigos 2017: Role of Metallic Nanoparticles in Vaccinology: Implications for Infectious Disease Vaccine Development.
Por Lázaro Moreira Marques Neto Em 19/05/2017 às 11:25

Abstract

Subunit vaccines are safer but less immunogenic than live-attenuated vaccines or whole cell inactivated vaccines. Adjuvants are used to enhance and modulate antigen (Ag) immunogenicity, aiming to induce a protective and long-lasting immune response. Several molecules and formulations have been studied for their adjuvanticity, but only seven have been approved to formulate human vaccines. Metallic nanoparticles (MeNPs), particularly those containing gold and iron oxides, are widely used in medicine for diagnosis and therapy and have been used as carriers for drugs and vaccines. However, little is known about the immune response elicited by MeNPs or about their importance in the development of new vaccines. There is evidence that these particles display adjuvant characteristics, promoting cell recruitment, antigen-presenting cell activation, cytokine production, and inducing a humoral immune response. This review focuses on the characteristics of MeNPs that could facilitate the induction of a cellular immune response, particularly T-helper 1 and T-helper 17, and their potential functions as adjuvants for subunit vaccines.

 

http://journal.frontiersin.org/article/10.3389/fimmu.2017.00239/full

massilience
Artigos 2017: Non-disulfide-Bridge Peptide 5.5 from the Scorpion Hadrurus gertschi Inhibits the Growth of Mycobacterium abscessus subsp. massiliense.
Por Lázaro Moreira Marques Neto Em 19/05/2017 às 11:22

Abstract

Multi-drug resistant microorganisms have been a growing concern during the last decades due to their contribution in mortality rates worldwide. Antimicrobial peptides (AMPs) are broad spectrum antimicrobial agents that display potent microbicidal activity against a wide range of microorganisms. AMPs generally have a rapid mode of action that reduces the risk of resistance developing among pathogens. In this study, an AMP derived from scorpion venom, NDBP-5.5, was evaluated against Mycobacterium abscessus subsp. massiliense, a rapidly growing and emerging pathogen associated with healthcare infections. The minimal bactericidal concentration of NDBP-5.5, AMP quantity necessary to stop bacteria visible growth, against M. abscessus subsp. massiliense was 200 μM, a concentration that did not induce hemolysis of human red blood cells. The therapeutic index was 3.05 indicating a drug with low toxicity and therefore good clinical potential. Treatment of infected macrophages with NDBP-5.5 or clarithromycin presented similar results, reducing the bacterial load. M. abscessus subsp. massiliense-infected animals showed a decrease in the bacterial load of up to 70% when treated with NDBP-5.5. These results revealed the effective microbicidal activity of NDBP-5.5 against Mycobacterium, indicating its potential as an antimycobacterial agent.

http://journal.frontiersin.org/article/10.3389/fmicb.2017.00273/full

Artigo 2017: Evaluation of the antimicrobial activity of the mastoparan Polybia-MPII isolated from venom of the social wasp Pseudopolybia vespiceps testacea (Vespidae, Hymenoptera).
Por Lázaro Moreira Marques Neto Em 19/05/2017 às 11:20

Abstract

Mastoparans, a class of peptides found in wasp venom, have significant effects following a sting as well as useful applications in clinical practice. Among these is their potential use in the control of micro-organisms that cause infectious diseases with a significant impact on society. Thus, the present study describes the isolation and identification of a mastoparan peptide from the venom of the social wasp Pseudopolybia vespiceps and evaluated its antimicrobial profile against bacteria (Staphylococcus aureus and Mycobacterium abscessus subsp. massiliense), fungi (Candida albicans and Cryptococcus neoformans) and in vivo S. aureus infection. The membrane pore-forming ability was also assessed. The mastoparan reduced in vitro and ex vivo mycobacterial growth by 80% at 12.5 µM in infected peritoneal macrophages but did not affect the shape of bacterial cells at the dose tested (6.25 µM). The peptide also showed potent action against S. aureus in vitro (EC50 and EC90 values of 1.83 µM and 2.90 µM, respectively) and reduced the in vivo bacterial load after 6 days of topical treatment (5 mg/kg). Antifungal activity was significant, with EC50 and EC90values of 12.9 µM and 15.3 µM, respectively, for C. albicans, and 11 µM and 22.70 µM, respectively, for C. neoformans. Peptides are currently attracting interest for their potential in the design of antimicrobial drugs, particularly due to the difficulty of micro-organisms in developing resistance to them. In this respect, Polybia-MPII proved to be highly effective, with a lower haemolysis rate compared with peptides of the same family.

 

https://linkinghub.elsevier.com/retrieve/pii/S0924-8579(17)30006-7

Defesas de TCC: Aline, Lucas Cacciari e Victor Procópio
Por Ana Paula Junqueira Kipnis Em 01/12/2016 às 16:48

Motivos de sobra para comemorar!

Parabéns a todos!

componentes
Os integrantes da Rede desejam a todos BOAS FESTAS!
Por Ana Paula Junqueira Kipnis Em 01/12/2016 às 16:32

Que Deus possa carregar nossos corações com boas vibrações!

SAIU O PRIMEIRO ARTIGO DOUTORADO DE MONALISA TRENTINI
Por Ana Paula Junqueira Kipnis Em 28/08/2016 às 21:09
Front Microbiol. 2016 Jun 10;7:898. doi: 10.3389/fmicb.2016.00898. eCollection 2016.

The Role of Neutrophils in the Induction of Specific Th1 and Th17 during Vaccination against Tuberculosis.

Abstract

Mycobacterium tuberculosis causes tuberculosis (TB), a disease that killed more than 1.5 million people worldwide in 2014, and the Bacillus Calmette Guérin (BCG) vaccine is the only currently available vaccine against TB. However, it does not protect adults. Th1 and Th17 cells are crucial for TB control, as well as the neutrophils that are directly involved in DC trafficking to the draining lymph nodes and the activation of T lymphocytes during infection. Although several studies have shown the importance of neutrophils during M. tuberculosis infection, none have shown its role in the development of a specific response to a vaccine. The vaccine mc(2)-CMX was shown to protect mice against M. tuberculosis challenge, mainly due to specific Th1 and Th17 cells. This study evaluated the importance of neutrophils in the generation of the Th1- and Th17-specific responses elicited by this vaccine. The vaccine injection induced a neutrophil rich lesion with a necrotic central area. The IL-17 KO mice did not generate vaccine-specific Th1 cells. The vaccinated IL-22 KO mice exhibited Th1- and Th17-specific responses. Neutrophil depletion during vaccination abrogated the induction of Th1-specific responses and prohibited the bacterial load reduction observed in the vaccinated animals. The results show, for the first time, the role of neutrophils in the generation of specific Th1 and Th17 cells in response to a tuberculosis vaccine.

KEYWORDS:

Th1/Th17 responses; neutrophils; tuberculosis vaccines

I Workshop do PGBB-Goiás
Por Ana Paula Junqueira Kipnis Em 10/05/2016 às 08:15

Para maiores informações acessar:

http://aurelionetorv.wix.com/workshopppgbb

V Workshop Nacional da Rede Brasileira de Pesquisas em Tuberculose – Rede TB,
Por Ana Paula Junqueira Kipnis Em 10/05/2016 às 08:14

ocorrerá em paralelo ao 52° Congresso da Sociedade Brasileira de Medicina Tropical no Centro de Convenções - Maceió/AL, de 22 a 24 de agosto de 2016.

Os inscritos no V Workshop Nacional da Rede estarão isentos da anuidade de 2016, tendo, entretanto, que quitar a anuidade de 2015. A  anuidade foi corrigida para R$ 150,00 para profissionais e R$ 80,00 para estudantes (dúvidas e informações: goudatb@gmail.com ). 

Os valores das inscrições do V Workshop da Rede TB serão os idênticos aos do 52° Congresso da Sociedade Brasileira de Medicina Tropical,  sendo que ao se inscrever em um deles terá acesso ao outro evento, sem custos adicionais.

http://eventos.livera.com.br/inscricao/?codEvento=145&lang=BR

O pagamento poderá ser efetuado no boleto ou parcelado nos cartões de crédito.

Teremos 12 Miniconferências, 20 mesas redondas, 3 simpósios, e 20 temas livres. Os Temas livres serão selecionados entre os melhores posters encaminhados ao Workshop. O programa científico está sendo finalizado, quando será definida a participação de palestrantes nacionais.

 

O prazo limite para submissão é 15 de junho de 2016.

 

logo
Publicações 2016 (II):Antimycobacterial Activity of a New Peptide Polydim-I Isolated from Neotropical Social Wasp Polybia dimorpha
Por Lázaro Moreira Marques Neto Em 08/03/2016 às 19:26

As micobacterioses também são de interesse de estudo para o nosso grupo de pesquisa, em especial as micobactérias

de crescimento rápido do grupo M. abscessus. Essas micobactérias, tem perfil de susceptibilidade a antimicrobianos e desinfetantes

diferenciado de outras bactérias do gênero e tem causado surto em todo o mundo, principalmente relacionado a pequenas cirurgias

ou procedimentos minimamente invasivos. Um dos maiores surtos brasileiros foi descrito e caracterizado com auxílio do nosso grupo de 

pesquisa e o artigo resultante do estudo das bactérias relacionadas a ele, no estado de Goiás, foi publicado na revista "Microbes and Infection" no ano de de 2008.

link:http://www.sciencedirect.com/science/article/pii/S1286457908002670

Desde então, nosso laboratório tem estudado essas micobacterioses e desde 2012 passamos a procurar alternativas terapêuticas para o tratamento de infecções causadas por essa bactéria. E foi na nossa fauna que encontramos compostos derivados da peçonha de vespas, em parceria com a rede INOVATOXIN, que são capazes de inibir o crescimento dessas bactérias.

Esse estudo é decorrente do projeto de mestrado do aluno Rogério Coutinho das Neves que com seus colaboradores acaba de publicar

os dados referentes ao seu trabalho. A publicação foi na revista "PlosOne" e tem título "Antimycobacterial Activity of a New Peptide Polydim-I Isolated from Neotropical Social Wasp Polybia dimorpha".

Segue abaixo o link da publicação, que já pode ser acessada online:

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0149729

logo
Publicações 2016 (I):The mc2-CMX vaccine induces an enhanced immune response against Mycobacterium tuberculosiscompared to BCG but with similar lung inflammatory effects
Por Lázaro Moreira Marques Neto Em 08/03/2016 às 19:14

Nossos trabalhos com vacina continuam rendendo frutos. A vacina recombinante MC2-CMX que vem sendo estudada

como uma vacina para tuberculose teve mais  uma trabalho publicado pelo grupo. Sua primeira publicação, de autoria

principal da Professora Doutora Ana Paula pode ser acessada pelo link: 

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0078639

Em complemento a esse trabalho, o aluno de doutorado Fábio Muniz de Oliveira e colaboradores acabam de ter seu trabalho

aceito na renomada revista brasileira "Memórias do Instituto Oswaldo Cruz", a melhor revista brasileira na área.

O trabalho tem financiamento da CNPq, CAPES, FAPEG.

Abaixo o resumo do trabalho, que ainda não teve sua  publicação online liberada.

Title: The mc2-CMX vaccine induces an enhanced immune response against Mycobacterium tuberculosiscompared to BCG but with similar lung inflammatory effects

Although the attenuated Mycobacterium bovis BCG vaccine has been used since 1921, tuberculosis control still proceeds at a slow pace. The main reason is the variable efficacy of BCG protection against tuberculosis among adults, which ranges from 0 to 80%. Subsequently, the mc2-CMX vaccine was developed, with promising results. Nonetheless, this recombinant vaccine needs to be compared to the standard BCG vaccine. The objective of this study was to evaluate the immune response induced by mc2-CMX and compare it to the response generated by BCG. BALB/c mice were immunized with both vaccines and challenged with M. tuberculosis. The immune and inflammatory responses were evaluated by ELISA, flow cytometry and histopathology.Mice vaccinated with mc2-CMX and challenged with Mycobacterium tuberculosis induced an increase in the IgG1 and IgG2 levels againstCMX as well as recalled specific CD4+ T cells that produced Th1 cytokines in the lungs and spleen compared with BCG vaccinated and challenged mice. Both vaccines reduced the lung inflammatory pathology induced by the M. tuberculosis infection. The mc2-CMX vaccine induces a humoral and cellular response that is superior to BCG and is efficiently recalled after challenge with M. tuberculosis, although both vaccines induced similar inflammatory reductions.

 

Convidamos a todos para a Defesa de Doutorado de Adeliane Castro da Costa
Por Ana Paula Junqueira Kipnis Em 01/03/2016 às 10:34

Título: Avaliação da Modulação da Resposta Imune induzida por BCG recombinante expressando epítopos imunodominantes de Mycobacterium tuberculosis.

Local: sala de Leitura, Horário: 13:00h

 

Foto Beatriz para pagina dos alunos
Defesa de TCC - Beatriz Cristina de Oliveira Fonseca
Por Lázaro Moreira Marques Neto Em 18/12/2015 às 09:49

No dia 17 de dezembro de 2015, a aluna Beatriz Cristina de Oliveira Fonseca, do laboratório de Microbiologia Molecular, defendeu o TCC de titulo "AVALIAÇÃO DO CRESCIMENTO IN VITRO DE Mycobacterium abscessus subsp. bolletii NA AUSÊNCIA E NA PRESENÇA DE FERRO E AVALIAÇÃO DA EXPRESSÃO DO GENE IDER, EM DIFERENTES CONCENTRAÇÕES DE FERRO E VARIAÇÕES DE PH." e foi aprovada pelo ótimo trabalho.

Parabéns pelo ótimo trabalho, Bia!

 

tcc beatriz