Abstract Two years ago, we held an exciting event entitled the São Paulo School of Advanced Sciences on Vaccines (SPSASV). Sixty-eight Ph.D. students, postdoctoral fellows and independent researchers from 37 different countries met at the Mendes Plaza Hotel located in the city of Santos, SP - Brazil to discuss the challenges and the new frontiers of vaccinology. The SPSASV provided a critical and comprehensive view of vaccine research from basics to the current state-of-the-art techniques performed worldwide. For 10 days, we discussed all the aspects of vaccine development in 36 lectures, 53 oral presentations and 2 poster sessions. At the end of the course, participants were further encouraged to present a model of a grant proposal related to vaccine development against individual pathogens. Among the targeted pathogens were viruses (Chikungunya, HIV, RSV, and Influenza), bacteria (Mycobacterium tuberculosis and Streptococcus pyogenes), parasites (Plasmodium falciparum or Plasmodium vivax), and the worm Strongyloides stercoralis. This report highlights some of the knowledge shared at the SPSASV.

Abstract Staphylococcus spp. play an important role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most relevant species due to the production of virulence factors such as slime, which is required for biofilm formation. This study aimed to evaluate biofilm production and its possible relation to beta-lactamic resistance in 20 S. aureus isolates from bovine mastitic milk. The isolates were characterized by pheno-genotypic and MALDI TOF-MS assays and tested for genes such as icaA, icaD, bap, agr RNAIII, agr I, agr II, agr III, and agr IV, which are related to slime production and its regulation. Biofilm production in microplates was evaluated considering the intervals determined along the bacterial growth curve. In addition, to determine the most suitable time interval for biofilm analysis, scanning electron microscopy was performed. Furthermore, genes such as mecA and blaZ that are related to beta-lactamic resistance and oxacillin susceptibility were tested. All the studied isolates were biofilm producers and mostly presented icaA and icaD. The Agr type II genes were significantly prevalent. According to the SEM, gradual changes in the bacterial arrangement were observed during biofilm formation along the growth curve phases, and the peak was reached at the stationary phase. In this study, the penicillin resistance was related to the production of beta-lactamase, and the high minimal bactericidal concentration for cefoxitin was possibly associated with biofilm protection. Therefore, further studies are warranted to better understand biofilm formation, possibly contributing to our knowledge about bacterial resistance in vivo.

Staphylococcus aureus antimicrobial resistance, especially to beta-lactams, favors treatment failures and its persistence in herd environment. This work aimed to develop a more specific primer for mecA gene detection based on the comparison of the conserved regions from distinct host origins and also investigated the presence of homologue mecA LGA251 in bovine strains. A total of 43 Staphylococcus spp. were included in this study, comprising 38 bovine S. aureus, two human and three equine coagulase-negative staphylococci (CNS). Phenotypical methicillin-resistance detection was performed through oxacillin agar-screening and cefoxitin disk-diffusion test. None isolate tested positive for mecA LGA251 gene. For mecA gene PCR, new primers were designed based on the sequences of human S. aureus (HE681097) and bovine S. sciuri (AY820253) mecA. The new primers based on the S. aureus mecA sequence amplified fragments of human and equine CNS and the ones based on S. sciuri mecA sequence only yielded fragments for S. aureus bovine strains. Multiples alignments of mecA gene sequences from bovine, human and equine revealed punctual but significant differences in bovine strains that can lead to the mecA gene detection impairment. The observed divergences of mecA gene sequences are not a matter of animal or human origin, it is a specificity of bovine samples.

Energy dispersive X-ray fluorescence methodology (EDXRF) was used to determine Al, Ba, Ca, Cr, Fe, K, Mn, Pb, Rb, S, Si, Sr, Ti, V, Zn in pottery sherds from seven archaeological sites in the central region of Rio Grande do Sul State, Brazil. The potteries' chemical fingerprints from Ijuí River, Ibicuí Mirim River, Vacacaí Mirim River and Jacuí River were identified. Interactions between sites from the Jacuí River, Vacacaí Mirim River and Ibicui Mirim River could have occurred because some samples from these sites are overlapping in a principal component analysis (PCA) graphic. The pottery provenance could be the same.

FUNDAMENTOS: A progressão linfática do melanoma maligno habitualmente se inicia pelo linfonodo sentinela (LNS), cuja análise histopatológica permite predizer o acometimento de toda a cadeia. OBJETIVO: O trabalho tem por objetivo descrever a utilização do 99mTc-Fitato na detecção do LNS em pacientes com melanoma maligno, revisando as indicações e informações fornecidas por sua biópsia. MÉTODO: A pesquisa de LNS foi realizada por meio da linfocintilografia com 99mTc-Fitato em 92 pacientes com melanoma (54,0±14,3 anos). Após 18-24 horas, 88 pacientes foram submetidos à localização intra-operatória com detector portátil, seguida da ressecção e análise histopatológica do LNS. RESULTADOS: A linfocintilografia permitiu a identificação do LNS em todos os estudos, havendo detecção intra-operatória em 98,8% dos casos. O LNS estava acometido em 23 pacientes (26%). O valor preditivo negativo foi de 100% e não se observaram reações adversas pelo uso do 99mTc-Fitato. CONCLUSÃO: A detecção do LNS pode ser realizada com diferentes radiofármacos, incluindo o 99mTc-Fitato, que apresenta vantagens de custo e disponibilidade no Brasil. A pesquisa de LNS resulta em maior acurácia e menor morbidade no estadiamento de pacientes com melanoma maligno

BACKGROUND: Sentinel lymph node (SLN), corresponding to the first lymph node draining the tumor, is usually the first one to receive its metastasis, and its biopsy is used to define the status of the whole lymphatic basin. OBJECTIVE: The aim of this paper is to describe the use 99mTc-Fitato in SLN localization in malignant melanoma patients, and to review the main indications and information provided by SLN biopsy. METHOD: A total of 92 patients with malignant melanoma was studied. Lymph node scintigraphy was carried out after the subdermal injection of 99mTc-Phytate. After 18-24 hours, intra-operative SLN localization was carried out using the gamma-probe and lymph node dissection was then performed. RESULTS: Lymphoscintigraphy identified the sentinel node in all studies and intra-operative detection using gamma-probe was reached in 98.8% of the cases. The SLN was involved in 23 patients (26%). The method's negative predictive value was 100%, and there were no side effects related to 99mTc-Phytate. CONCLUSION: Scintigraphic and intra-operative sentinel node detection was satisfactorily performed using 99mTc-Phytate, an easily available and low cost radiopharmaceutical. SLN mapping allows the use of more accurate tumor staging techniques and reduces surgical morbidity.

Parasitas intracelulares obrigatórios como Plasmodium sp, Trypanosoma cruzi, Toxoplasma gondii e Leishmania sp são responsáveis pela infecção de milhões de indivíduos a cada ano. Estes parasitas são capazes de liberar antígenos no citoplasma de células infectadas do hospedeiro que são apresentados por moléculas de MHC classe I para células T CD8 protetoras. As condições de estímulo in vivo destas células T CD8 específicas durante a infecção natural são pouco conhecidas e constituem uma área pouco explorada. Os mecanismos anti-parasitários mediados por células T CD8 incluem vias dependentes e independentes do interferon-g. O fato que células T CD8 são potentes inibidores do desenvolvimento parasitário levou diversos investigadores a explorarem se a indução destes linfócitos T poderia constituir uma estratégia factível para o desenvolvimento de vacinas efetivas contra estas doenças parasitárias. Estudos feitos em modelos experimentais suportam a hipótese que células T CD8 induzidas por vetores recombinantes virais ou vacinas de DNA podem servir de base para a vacinação humana. Regimes de imunização consistindo de dois vetores distintos (prime-boost heterólogo) são muito mais eficientes em termos da expansão de linfócitos T CD8 protetores do que a imunização com um único vetor. Os resultados obtidos usando modelos experimentais levaram a vacinações clínicas que estão atualmente em curso.

Obligatory intracellular parasites such as Plasmodium sp, Trypanosoma cruzi, Toxoplasma gondii and Leishmania sp are responsible for the infection of hundreds of millions of individuals every year. These parasites can deliver antigens to the host cell cytoplasm that are presented through MHC class I molecules to protective CD8 T cells. The in vivo priming conditions of specific CD8 T cells during natural infection are largely unknown and remain as an area that has been poorly explored. The antiparasitic mechanisms mediated by CD8 T cells include both interferon-g-dependent and -independent pathways. The fact that CD8 T cells are potent inhibitors of parasitic development prompted many investigators to explore whether induction of these T cells can be a feasible strategy for the development of effective subunit vaccines against these parasitic diseases. Studies performed on experimental models supported the hypothesis that CD8 T cells induced by recombinant viral vectors or DNA vaccines could serve as the basis for human vaccination. Regimens of immunization consisting of two different vectors (heterologous prime-boost) are much more efficient in terms of expansion of protective CD8 T lymphocytes than immunization with a single vector. The results obtained using experimental models have led to clinical vaccination trials that are currently underway.