Impactos do pisoteio humano na fauna de um costão rochoso do litoral de São Paulo, no sudeste brasileiro

Increased tourist activity in coastal regions demands management strategies to reduce impacts on rocky shores. The highly populated coastal areas in southeastern Brazil are an example of degradation caused by development of industry and tourism. Among different shore impacts, trampling has been intensively studied, and may represent a significant source of stress for intertidal fauna. A randomised blocks design was applied to experimentally study the effects of two different trampling intensities on richness, diversity, density and biomass of the rocky shore fauna of Obuseiro beach, Guarujá, southeastern Brazil. Blocks were distributed in two portions of the intertidal zone, dominated respectively by Chthamalus bisinuatus (Cirripedia) and Isognomon bicolor (Bivalvia). Blocks were trampled over three months, simulating the vacation period in Brazil and were monitored for the following nine months. Results indicate that Chthamalus bisinuatus is vulnerable to trampling impacts. Richness, diversity and turn-over index tended to be higher in trampled plots four months after trampling ceased. In general, results agree with previous trampling studies, suggesting that even low intensities of trampling may cause some impact on intertidal communities. Management strategies should include isolation of sensitive areas, construction of boardwalks, visitor education and monitoring programmes. In Brazil, additional data obtained from experimental studies are necessary in order to achieve a better understanding of trampling impacts on rocky shore communities.O aumento da atividade turística em áreas costeiras nas últimas décadas faz necessária a adoção de estratégias de manejo para reduzir os impactos gerados às comunidades de costões rochosos. A região costeira do sudeste brasileiro possui bons exemplos de degradação causada pelo turismo e desenvolvimento industrial. Dentre os diferentes distúrbios causados pela visitação, o pisoteio têm sido estudado de forma intensa e pode representar uma fonte significativa de impactos para as comunidades da zona entre-marés. Neste projeto, foi aplicado um desenho de blocos randômicos para se avaliar experimentalmente os efeitos de duas intensidades de pisoteio na riqueza, diversidade, densidade, recobrimento e biomassa da fauna de um costão situado na praia do Obuseiro, no município do Guarujá, São Paulo, Brasil. Os blocos foram alocados em dois povoamentos diferentes, dominados respectivamente por Chthamalus bisinuatus (Cirripedia) e Isognomon bicolor (Bivalvia). O pisoteio foi aplicado durante três meses, simulando a temporada de férias no Brasil, e os blocos foram monitorados nos nove meses seguintes. Os resultados indicaram que Chthamalus bisinuatus é vulnerável aos impactos do pisoteio. Os índices de riqueza, diversidade e turnover apresentaram uma tendência ao aumento nas áreas pisoteadas quatro meses após o término do pisoteio. No geral, os resultados concordam com estudos anteriores e sugerem que mesmo baixas intensidades do distúrbio podem causar impacto às comunidades bentônicas. Estratégias de manejo devem envolver o isolamento de áreas sensíveis, a construção de passarelas, a educação dos visitantes e o monitoramento das comunidades impactadas. No Brasil, a realização de maior quantidade de estudos experimentais é necessária para a melhor compreensão dos impactos do pisoteio nas comunidades de costões rochosos

Haemophilia B Curative FIX Production from a Low Dose UCOE-based Lentiviral Vector Following Hepatic Pre-natal Delivery

The ubiquitous chromatin opening element from the human HNRPA2B1-CBX3 housekeeping gene locus (A2UCOE) is able to provide stable and cell-to-cell reproducible levels of transgene expression regardless of target cell genome integration site with efficacy demonstrated in adult, embryonic and induced pluripotent stem cells and their differentiated progeny in vitro and in vivo. Here we evaluate the ability of A2UCOE-based lentiviral vectors to confer stable expression following pre-natal delivery in mice. Our results show stable post-natal A2UCOE-eGFP and A2UCOE-luciferase lentiviral vector presence in both the liver and haematopoietic system with concomitant persistence of expression demonstrating efficient transduction of both fetal liver and haematopoietic stem cells. In addition, we find that an A2UCOE-FIX lentiviral vector produces comparable amounts of plasma FIX protein to that obtained from a SFFV-FIX construct. Furthermore, the A2UCOE-FIX vector shows that at a low (0.19) average vector copy number per liver cell, it can provide stable levels of plasma FIX production, which would convert severe haemophilia B (<1%) to a mild phenotype (≈20%). Our results provide proof-ofprinciple for low dose pre-natal A2UCOE-based LV delivery to the liver as a therapeutic option for haemophilia B and potentially other metabolic conditions

Critical evaluation of proteomic protocols for passion fruit (Passiflora edulis Sims) leaves, a crop with juice market benefits

Passion fruit grows practically all over Brazilian territory; its production is largely destined to juice industry and expanding to overseas markets. The suitability of four protein extraction protocols for plant proteome was  investigated to determine the best choice for studies concerning passion fruit leaf proteins. Trichloroacetic acid (TCA)/acetone extraction; isoelectric  focusing (IEF) buffer extraction; phenol (Phe) extraction and Phe-SDS extraction were tested. The Phe method produced the best results, showing higher reproducibility of resolved protein spots and clearer 2D gel  background staining. In comparison, the Phe-SDS method presented fewer spots and lower reproducibility. The TCA/acetone method produced the fewest identifiable spots and the IEF buffer produced the poorest results,displaying fewer reproducibly detected spots, more vertical streaks and darker 2D staining. Selected spots, obtained with Phe method, were identified by spectrometric analysis (MALDI-TOF-TOF) to exemplify the viability to perform more comprehensive proteomic studies with passion fruit leaves and, therefore increase information about stress-related and developmental responses in this fruit crop.Key words: Passion fruit, proteomic, protein extraction, juice industry

Near full-length genome analysis of low prevalent human immunodeficiency virus type 1 subclade F1 in São Paulo, Brazil

Background: The genetic diversity of the human immunodeficiency virus type 1 (HIV-1) is critical to lay the groundwork for the design of successful drugs or vaccine. In this study we aimed to characterize and define the molecular prevalence of HIV-1 subclade F1 currently circulating in Sao Paulo, Brazil. Methods: A total of 36 samples were selected from 888 adult patients residing in Sao Paulo who had previously been diagnosed in two independent studies in our laboratory as being infected with subclade F1 based on pol subgenomic fragment sequencing. Proviral DNA was amplified from the purified genomic DNA of all 36 blood samples by 5 fragments overlapping PCR followed by direct sequencing. Sequence data were obtained from the 5 fragments of pure subclade F1 and phylogenetic trees were constructed and compared with previously published sequences. Subclades F1 that exhibited mosaic structure with other subtypes were omitted from any further analysis Results: Our methods of fragment amplification and sequencing confirmed that only 5 sequences inferred from pol region as subclade F1 also holds true for the genome as a whole and, thus, estimated the true prevalence at 0.56%. The results also showed a single phylogenetic cluster of the Brazilian subclade F1 along with non-Brazilian South American isolates in both subgenomic and the full-length genomes analysis with an overall intrasubtype nucleotide divergence of 6.9%. The nucleotide differences within the South American and Central African F1 strains, in the C2-C3 env, were 8.5% and 12.3%, respectively. Conclusion: All together, our findings showed a surprisingly low prevalence rate of subclade F1 in Brazil and suggest that these isolates originated in Central Africa and subsequently introduced to South America.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[06/50096-0

Effect of hyperbaric stress on yeast morphology: Study by automated image analysis

The effects of hyperbaric stress on the morphology of Saccharomyces cerevisiae were studied in batch cultures under pressures between 0.1 MPa and 0.6 MPa and different gas compositions (air, oxygen, nitrogen or carbon dioxide), covering aerobic and anaerobic conditions. A method using automatic image analysis for classification of S. cerevisiae cells based on their morphology was developed and applied to experimental data. Information on cell size distribution and bud formation throughout the cell cycle is reported. The results show that the effect of pressure on cell activity strongly depends on the nature of the gas used for pressurization. While nitrogen and air to a maximum of 0.6 MPa of pressure were innocuous to yeast, oxygen and carbon dioxide pressure caused cell inactivation, which was confirmed by the reduction of bud cells with time. Moreover, a decrease in the average cell size was found for cells exposed for 7.5 h to 0.6 MPa CO2.CAPES and CNPq (Brazil). Fundação para a Ciência e Tecnologia (Portugal)

Intact triacylglycerol profiles of fats and meats via thermal imprinting easy ambient sonic-spray ionization mass spectrometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Thermal imprinting (TI) on a paper surface, using minimal solvent amounts, followed by direct analysis of the triacylglycerols (TAG) content via easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is shown to provide a fast protocol to analyze TAG in meats and fats. The technique is simple, fast and eco-friendly requiring no hydrolysis, derivatization or chromatographic separation. The entire TI-EASI-MS protocol is performed in a few minutes and with minimal sample handling and solvent consumption. The TAG profiles obtained via TI-EASI-MS are shown to be quite similar to those obtained using GC and MALDI-MS analyses, and the imprinting and mailing of the imprinted paper in a sealed plastic bag is proposed for remote TI-EASI-MS analysis of meat and fat samples.41135513557Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Financing Agency of Studies and Projects (FINEP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Target Score-A Proteomics Data Selection Tool Applied to Esophageal Cancer Identifies GLUT1-Sialyl Tn Glycoforms as Biomarkers of Cancer Aggressiveness

Esophageal cancer (EC) is a life-threatening disease, demanding the discovery of new biomarkers and molecular targets for precision oncology. Aberrantly glycosylated proteins hold tremendous potential towards this objective. In the current study, a series of esophageal squamous cell carcinomas (ESCC) and EC-derived circulating tumor cells (CTCs) were screened by immunoassays for the sialyl-Tn (STn) antigen, a glycan rarely expressed in healthy tissues and widely observed in aggressive gastrointestinal cancers. An ESCC cell model was glycoengineered to express STn and characterized in relation to cell proliferation and invasion in vitro. STn was found to be widely present in ESCC (70% of tumors) and in CTCs in 20% of patients, being associated with general recurrence and reduced survival. Furthermore, STn expression in ESCC cells increased invasion in vitro, while reducing cancer cells proliferation. In parallel, an ESCC mass spectrometry-based proteomics dataset, obtained from the PRIDE database, was comprehensively interrogated for abnormally glycosylated proteins. Data integration with the Target Score, an algorithm developed in-house, pinpointed the glucose transporter type 1 (GLUT1) as a biomarker of poor prognosis. GLUT1-STn glycoproteoforms were latter identified in tumor tissues in patients facing worst prognosis. Furthermore, healthy human tissues analysis suggested that STn glycosylation provided cancer specificity to GLUT1. In conclusion, STn is a biomarker of worst prognosis in EC and GLUT1-STn glycoforms may be used to increase its specificity on the stratification and targeting of aggressive ESCC forms