Bipolar androgen therapy plus nivolumab for patients with metastatic castration-resistant prostate cancer: the COMBAT phase II trial.

Cyclic high-dose testosterone administration, known as bipolar androgen therapy (BAT), is a treatment strategy for patients with metastatic castration-resistant prostate cancer (mCRPC). Here, we report the results of a multicenter, single arm Phase 2 study (NCT03554317) enrolling 45 patients with heavily pretreated mCRPC who received BAT (testosterone cypionate, 400 mg intramuscularly every 28 days) with the addition of nivolumab (480 mg intravenously every 28 days) following three cycles of BAT monotherapy. The primary endpoint of a confirmed PSA50 response rate was met and estimated at 40% (N = 18/45, 95% CI: 25.7-55.7%, P = 0.02 one-sided against the 25% null hypothesis). Sixteen of the PSA50 responses were achieved before the addition of nivolumab. Secondary endpoints included objective response rate (ORR), median PSA progression-free survival, radiographic progression-free survival (rPFS), overall survival (OS), and safety/tolerability. The ORR was 24% (N = 10/42). Three of the objective responses occurred following the addition of nivolumab. After a median follow-up of 17.9 months, the median rPFS was 5.6 (95% CI: 5.4-6.8) months, and median OS was 24.4 (95% CI: 17.6-31.1) months. BAT/nivolumab was well tolerated, resulting in only five (11%) drug related, grade-3 adverse events. In a predefined exploratory analysis, clinical response rates correlated with increased baseline levels of intratumoral PD-1 + T cells. In paired metastatic tumor biopsies, BAT induced pro-inflammatory gene expression changes that were restricted to patients achieving a clinical response. These data suggest that BAT may augment antitumor immune responses that are further potentiated by immune checkpoint blockade.

Chromogenic detection of telomere lengths in situ aids the identification of precancerous lesions in the prostate.

BACKGROUND: Telomeres are terminal chromosomal elements that are essential for the maintenance of genomic integrity. The measurement of telomere content provides useful diagnostic and prognostic information, and fluorescent methods have been developed for this purpose. However, fluorescent-based tissue assays are cumbersome for investigators to undertake, both in research and clinical settings. METHODS: A robust chromogenic in situ hybridization (CISH) approach was developed to visualize and quantify telomere content at single cell resolution in human prostate tissues, both frozen and formalin-fixed, paraffin-embedded (FFPE). RESULTS: This new assay (telomere chromogenic in situ hybridization ["Telo-CISH"]) produces permanently stained slides that are viewable with a standard light microscope, thus avoiding the need for specialized equipment and storage. The assay is compatible with standard immunohistochemistry, thereby allowing simultaneous assessment of histomorphology, identification of specific cell types, and assessment of telomere status. In addition, Telo-CISH eliminates the problem of autofluorescent interference that frequently occurs with fluorescent-based methods. Using this new assay, we demonstrate successful application of Telo-CISH to help identify precancerous lesions in the prostate by the presence of markedly short telomeres specifically in the luminal epithelial cells. CONCLUSIONS: In summary, with fewer restrictions on the types of tissues that can be tested, and increased histologic information provided, the advantages presented by this novel chromogenic assay should extend the applicability of tissue-based telomere length assessment in research and clinical settings.

Proteomic analysis of Cryptostegia grandiflora latex, purification, characterization, and biological activity of two osmotin isoforms.

Although latex fluids are found in >20,000 plant species, the biochemical composition and biological function of their proteins are still poorly explored. Thus, this work aimed to conduct a proteomic analysis of Cryptostegia grandiflora latex (CgLP) for subsequent purification and characterization of an antifungal protein. After 2D-SDS-PAGE and mass spectrometry, 27 proteins were identified in CgLP, including a polygalacturonase inhibitor, cysteine peptidases, pathogenesis-related proteins (PR-4), and osmotins. Then, two osmotin isoforms (CgOsm) were purified, and a unique N-terminal sequence was determined (1ATFDIRSNCPYTVWAAAVPGGGRRLDRGQTWTINVAPGTA40). The PCR products revealed a cDNA sequence of 609 nucleotides for CgOsm, which encoded a polypeptide with 203 amino acid residues. The structure of CgOsm has features of typical osmotin or thaumatin-like proteins (TLPs), such as 16 conserved Cys residues, REDDD and FF motifs, an acidic cleft, and three main domains. Atomic force microscopy (AFM) and bioinformatics suggested that CgOsm is associated with three chain units. This result was interesting since the literature describes osmotins and TLPs as monomers. AFM also showed that Fusarium falciforme spores treated with CgOsm were drastically damaged. Therefore, it is speculated that CgOsm forms pores in the membrane of these cells, causing the leakage of cytoplasmic content.

Sexually transmitted infections lesions found during colonoscopies

Introduction: Anal examination and videoanoscopy (VA) are rarely performed during colonoscopies. In recent years, there has been a considerable increase in lesions of sexually transmitted anal and rectal infections, but these conditions are not noticed or reported during routine colonoscopy. Objective: To raise awareness regarding the fortuitous findings of lesions and sexually transmitted infections (STIs) in colonoscopy exams and to demonstrate that anal examination and VA provide important information and should be routinely performed. Methods: We conducted a descriptive retrospective study in 16,132 patients screened by colonoscopy and VA between 2006 and 2018. Among numerous other findings, the presence of anal condylomata and sexually transmitted retitis or perianal dermatitis was observed. The rates of each finding were calculated, and the patients were subdivided by sex and into age groups by blocks of ten years. Results: Among the 16,132 colonoscopies performed, 26 cases of condyloma (0.16%) and 50 cases of proctitis or perianal dermatitis suspicious for STI (0.33%) were found. Conclusion: Performing anal examination and VA systematically in all routine colonoscopies enabled the identification of numerous anal conditions, including several fortuitous cases of STIs. The study proposes that anal examination and VA should be performed in all routine colonoscopies and, in suspected cases, complementary tests for STIs. (AU)

Chromogenic detection of telomere lengths in situ aids the identification of precancerous lesions in the prostate.

Telomeres are terminal chromosomal elements that are essential for the maintenance of genomic integrity. The measurement of telomere content provides useful diagnostic and prognostic information, and fluorescent methods have been developed for this purpose. However, fluorescent-based tissue assays are cumbersome for investigators to undertake, both in research and clinical settings. Here, a robust chromogenic in situ hybridization (CISH) approach was developed to visualize and quantify telomere content at single cell resolution in human prostate tissues, both frozen and formalin-fixed, paraffin-embedded (FFPE). This new assay ("Telo-CISH") produces permanently stained slides that are viewable with a standard light microscope, thus avoiding the need for specialized equipment and storage. The assay is compatible with standard immunohistochemistry, thereby allowing simultaneous assessment of histomorphology, identification of specific cell types, and assessment of telomere status. In addition, Telo-CISH eliminates the problem of autofluorescent interference that frequently occurs with fluorescent-based methods. Using this new assay, we demonstrate successful application of Telo-CISH to help identify precancerous lesions in the prostate by the presence of markedly short telomeres specifically in the luminal epithelial cells. In summary, with fewer restrictions on the types of tissues that can be tested, and increased histologic information provided, the advantages presented by this novel chromogenic assay should extend the applicability of tissue-based telomere length assessment in research and clinical settings.

Androgen receptor activity in prostate cancer dictates efficacy of bipolar androgen therapy through MYC.

Testosterone is the canonical growth factor of prostate cancer but can paradoxically suppress its growth when present at supraphysiological levels. We have previously demonstrated that the cyclical administration of supraphysiological androgen (SPA), termed bipolar androgen therapy (BAT), can result in tumor regression and clinical benefit for patients with castration-resistant prostate cancer. However, predictors and mechanisms of response and resistance have been ill defined. Here, we show that growth inhibition of prostate cancer models by SPA required high androgen receptor (AR) activity and were driven in part by downregulation of MYC. Using matched sequential patient biopsies, we show that high pretreatment AR activity predicted downregulation of MYC, improved clinical response, and prolonged progression-free and overall survival for patients on BAT. BAT induced strong downregulation of AR in all patients, which is shown to be a primary mechanism of acquired resistance to SPA. Acquired resistance was overcome by alternating SPA with the AR inhibitor enzalutamide, which induced adaptive upregulation of AR and resensitized prostate cancer to SPA. This work identifies high AR activity as a predictive biomarker of response to BAT and supports a treatment paradigm for prostate cancer involving alternating between AR inhibition and activation.

Overexpression of estrogen receptor GPER1 and G1 treatment reduces SARS-CoV-2 infection in BEAS-2B bronchial cells.

Gender-bias in COVID-19 severity has been suggested by clinical data. Experimental data in cell and animal models have demonstrated the role of sex hormones, particularly estrogens, in viral infections such as in COVID-19. SARS-CoV-2 uses ACE2 as a receptor to recognize host cells, and the protease TMPRSS2 for priming the Spike protein, facilitating virus entry into cells. However, the involvement of estrogenic receptors in SARS-CoV-2 infection are still being explored. Thus, in order to investigate the role of estrogen and its receptors in COVID-19, the estrogen receptors ERα, ERß and GPER1 were overexpressed in bronchial BEAS-2B cell, and then infected with SARS-CoV-2. Interestingly, the levels of ACE2 and TMPRSS2 mRNA were higher in SARS-CoV-2-infected cells, but no difference was observed in cells with estrogen receptors overexpression. GPER1 can be involved in virus infection or replication, since its higher levels reduces SARS-CoV-2 load. On the other hand, pharmacological antagonism of GPER1 enhanced viral load. Those data suggest that GPER1 has an important role in SARS-CoV-2 infection.

Multiplex immunohistochemical phenotyping of T cells in primary prostate cancer.

BACKGROUND: Most prostate cancers are "immune cold" and poorly responsive to immune checkpoint inhibitors. However, the mechanisms responsible for the lack of a robust antitumor adaptive immune response in the prostate are poorly understood, which hinders the development of novel immunotherapeutic approaches. AIMS: Most inflammatory infiltrates in the prostate are centered around benign glands and stroma, which can confound the molecular characterization of the antitumor immune response. We sought to analytically validate a chromogenic-based multiplex immunohistochemistry (IHC) approach applicable to whole slide digital image analysis to quantify T cell subsets from the tumor microenvironment of primary prostatic adenocarcinomas. As an initial application, we tested the hypothesis that PTEN loss leads to an altered antitumor immune response by comparing matched regions of tumors within the same individual with and without PTEN loss. MATERIALS & METHODS: Using the HALO Image Analysis Platform (Indica Labs), we trained a classifier to quantify the densities of eight T cell phenotypes separately in the tumor epithelial and stromal subcompartments. RESULTS: The iterative chromogenic approach using 7 different antibodies on the same slide provides highly similar findings to results using individually stained slides with single antibodies. Our main findings in carcinomas (benign removed) include the following: i) CD4+ T cells are present at higher density than CD8+ T cells; ii) all T cell subsets are present at higher densities in the stromal compartment compared to the epithelial tumor compartment; iii) most CD4+ and CD8+ T cells are PD1+; iv) cancer foci with PTEN loss harbored increased numbers of T cells compared to regions without PTEN loss, in both stromal and epithelial compartments; and v) the increases in T cells in PTEN loss regions were associated with ERG gene fusion status. DISCUSSION: This modular approach can apply to any IHC-validated antibody combination and sets the groundwork for more detailed spatial analyses. CONCLUSION: Iterative chromogenic IHC can be used for whole slide analysis of prostate tissue samples and can complement transcriptomic results including those using single cell and spatial genomic approaches.

Do monitoramento autônomo à pesquisa colaborativa virtual: parceria com o movimento indígena do Nordeste durante a pandemia da covid-19 como apoio ao controle social / From autonomous monitoring to virtual collaborative research: partnership with the indigenous movement from the Northeast region during covid-19 pandemic as support for social control

Resumo Nos primeiros meses da pandemia de covid-19, em 2020, os movimentos e organizações indígenas da região Nordeste do Brasil estabeleceram uma extensa rede de apoio e parcerias com grupos de pesquisadores e entidades da sociedade civil para a organização de campanhas de solidariedade aos povos indígenas. A produção de informações gerais e dados empíricos sobre como a doença atingiu os territórios e populações indígenas constituiu uma das principais estratégias de ação. Essa mobilização foi a base para a constituição de redes colaborativas que investigaram como ocorreu o enfrentamento dos povos diante da pandemia, por meio de um viés antropológico e aplicando métodos que poderíamos definir como uma pesquisa colaborativa virtual. Este artigo, portanto, discute o potencial desse tipo de parceria para a reflexão sobre o Subsistema de Atenção à Saúde Indígena, argumentando que esse modelo pode constituir uma forma de apoio ao controle social exercido por parte das comunidades.

Abstract In the first months of the covid-19 pandemic, in 2020, indigenous movements and organizations in the Northeast region of Brazil established an extensive network of support and partnerships with groups of researchers and civil society entities to organize campaigns of solidarity with the indigenous peoples. The production of general information and empirical data on how the disease reached indigenous territories and populations constituted one of the main strategies for action. This mobilization was the basis for establishing collaborative networks that investigated how the indigenous peoples faced the pandemic, from an anthropological bias and applying methods that we could define as virtual collaborative research. The article, thus, discusses the potential of this type of partnership for reflection on the Indigenous Health Care Subsystem, arguing that this model can constitute a kind of support for social control exercised by the communities.

Programa de Mentoria do Curso de Medicina da Universidade Federal do Rio Grande do Norte: Atividades Integrativas em Foco / The Universidade Federal do Rio Grande do Norte Medicine Course Mentoring Program: Integrative Activities in Focus

Resumo: Introdução: Ao ingressarem no curso de Medicina, os estudantes se deparam com desafios inerentes à formação que podem ser geradores de estresse e ansiedade, comprometendo seu bem-estar e desempenho acadêmico. Diante disso, o curso de Medicina da Universidade Federal do Rio Grande do Norte (UFRN) implantou em 2015 um programa de mentoria que se propõe a contribuir para o desenvolvimento profissional e pessoal do estudante, adotando-se como um diferencial a realização de atividades integrativas. Método: Participam desse programa 25 professores do curso de Medicina da UFRN, que atuam como mentores, 25 monitores ou mentores juniores, que têm o papel de intermediar a comunicação entre os participantes, como também auxiliar no planejamento e desenvolvimento das atividades, e 317 alunos de diversos períodos do curso médico. Além dos encontros mensais regulares, no final de cada semestre, os mentores e mentores juniores organizam a atividade integrativa que agrega todos os discentes e docentes do programa e possibilitam a construção de uma relação mais próxima entre mentores e mentorandos, bem como contribuem para a estruturação de um ambiente universitário mais acolhedor e equânime. Resultado: Em avaliação on-line sobre o programa, os alunos destacaram a "troca de experiências, de sugestões e a ajuda no curso sobre diversos temas" e "adquirir experiência de um profissional experiente" como principais motivações para participar da atividade. Contudo, enfatizaram a falta de tempo para conciliar os encontros com as demais atividades acadêmicas como principal obstáculo para participar do programa. Com a pandemia da Covid-19, mantiveram-se as atividades do programa no modelo on-line, e obteve-se importante engajamento dos participantes, o que representou uma estratégia de enfrentamento do isolamento social e de promoção da saúde mental para os estudantes. Conclusão: Observamos, a partir do engajamento e dos feedbacks recebidos, que o programa, apesar de apresentar alguns desafios, vem se configurando como uma iniciativa capaz de transformar as relações interpessoais entre discentes e mentores, ao promover a integração entre alunos dos diferentes períodos do curso e criar um ambiente favorável ao diálogo e à construção do conhecimento.

Abstract: Introduction: Upon starting their medical course, students are faced with challenges inherent to training, which can generate stress and anxiety, compromising their well-being and academic performance. In view of this, in 2015 the Medicine course at the Federal University of Rio Grande do Norte (UFRN) implemented a Mentoring Program, which aims to contribute to students' professional and personal development, focusing on integrative activities to strengthen bonds between the participants. Method: This program involves the participation of 25 professors from the UFRN Medicine course, who act as mentors, 25 monitors or mentors, who have the role of mediating communication between participants and assisting in the planning and development of activities; and about 317 students and various academic periods of the medical course. In addition to the regular monthly meetings, at the end of each semester, mentors and junior mentors organize the integrative activity that brings together all the students and teachers of the program and have stimulated a closer relationship between mentors and mentees, as well as contributing toward the structuring of a more welcoming and equitable university environment. Result: In an online assessment of the program, students highlighted the "exchange of experiences, suggestions and help with the course on various topics" and "acquiring experience from an experienced professional" as the main motivations for participating in the activity. On the other hand, they emphasized the lack of time to reconcile meetings with other academic activities as the main obstacle to participating in the program. With the onset of the Covid-19 pandemic, the program activities continued online and important engagement was achieved among the participants, thus constituting a strategy to face social isolation and promote the mental health of undergraduate medical students. Conclusion: From the engagement and the feedback received, it was observed that the program, despite presenting some challenges, represents an initiative capable of transforming interpersonal relationships between students and mentors, promoting integration between students from different academic periods of the course and creating a favourable environment for dialogue and knowledge construction.

Video anoscopy: results of routine anal examination during colonoscopies.

Background and study aims Anal examination and video anoscopy (VA) are rarely performed during colonoscopies. The aim of this study is to demonstrate that anal examination and VA provide important information in all routine colonoscopies. Patients and methods A cross-sectional study was conducted on 12,151 patients screened by VA which were performed during routine outpatient colonoscopy between 2006 and 2018. The aspects studied were: normal examination; hemorrhoidal disease; thrombosed hemorrhoids; anal fissure; perianal Crohn's Disease; perianal fistula; condyloma; polyps; neoplasms; stenosis; bleeding. Results Of the colonoscopies performed on 12,151 patients, 9,364 cases (77.06 %) presented some alterations. Internal I degree hemorrhoids (5911 - 48.65 %); internal hemorrhoids of II, III and IV degrees (2362 - 19.44 %); thrombosed hemorrhoids (120 - 0.99 %); bleeding (56 - 0.46 %); fissure (415 - 3.42 %); perianal fistula (42 - 0.35 %); perianal Crohn's Disease (34 - 0.28 %); condylomas (18 - 0.15 %); anal stenosis (30 - 0.25 %); other findings (310 - 2.55 %); polyps (62 - 0.51 %), one of which was adenomatous with high-grade dysplasia; four cases of anal canal neoplasia (0.03 %): two cases of squamous cell carcinoma and two cases of adenocarcinoma. Conclusion The association of routine video anoscopy during colonoscopy improved diagnosis of neoplastic anal lesions, allowed correct classification of the degree of hemorrhoidal disease, helped to confirm the bleeding site and detected other anal pathologies. The main findings were hemorrhoidal diseases, fissures and perianal fistulas. The study suggests that VA should be used in all colonoscopies.

Combining discovery and targeted proteomics reveals a prognostic signature in oral cancer.

Different regions of oral squamous cell carcinoma (OSCC) have particular histopathological and molecular characteristics limiting the standard tumor-node-metastasis prognosis classification. Therefore, defining biological signatures that allow assessing the prognostic outcomes for OSCC patients would be of great clinical significance. Using histopathology-guided discovery proteomics, we analyze neoplastic islands and stroma from the invasive tumor front (ITF) and inner tumor to identify differentially expressed proteins. Potential signature proteins are prioritized and further investigated by immunohistochemistry (IHC) and targeted proteomics. IHC indicates low expression of cystatin-B in neoplastic islands from the ITF as an independent marker for local recurrence. Targeted proteomics analysis of the prioritized proteins in saliva, combined with machine-learning methods, highlights a peptide-based signature as the most powerful predictor to distinguish patients with and without lymph node metastasis. In summary, we identify a robust signature, which may enhance prognostic decisions in OSCC and better guide treatment to reduce tumor recurrence or lymph node metastasis.

CONVENTIONAL VIDEOENDOSCOPY CAN IDENTIFY HELICOBACTER PYLORI GASTRITIS?

BACKGROUND: Studies with latest technologies such as endoscopy with magnification and chromoendoscopy showed that various endoscopic aspects are clearly related to infection by Helicobacter pylori (HP). The description of different patterns of erythema in gastric body under magnification of images revived interest in identifying these patterns by standard endoscopy. AIM: To validate the morphologic features of gastric mucosa related to H. pylori infection gastritis allowing predictability of their diagnosis as well as proper targeting biopsies. METHODS: Prospective study of 339 consecutive patients with the standard videoendoscope image analysis were obtained, recorded and stored in a program database. These images were studied with respect to the presence or absence of H. pylori, diagnosed by rapid urease test and/or by histological analysis. Were studied: a) normal mucosa appearance; b) mucosal nodularity; c) diffuse nonspecific erythema or redness (with or without edema of folds and exudate) of antrum and body; d) mosaic pattern with focal area of hyperemia; e) erythema in streaks or bands (red streak); f) elevated (raised) erosion; g) flat erosions; h) fundic gland polyps. The main exclusion criteria were the use of drugs, HP pre-treatment and other entities that could affect results. RESULTS: Applying the exclusion criteria, were included 170 of the 339 patients, of which 52 (30.58%) were positive for HP and 118 negative. On the positive findings, the most associated with infection were: nodularity in the antrum (26.92%); presence of raised erosion (15.38%) and mosaic mucosa in the body (21.15%). On the negative group the normal appearance of the mucosa was 66.94%; erythema in streaks or bands in 9.32%; flat erosions 11.86%; and fundic gland polyps 11.86%. CONCLUSION: Endoscopic findings are useful in the predictability of the result and in directing biopsies. The most representative form of HP related gastritis was the nodularity of the antral mucosa. The raised erosion and mucosa in mosaic in the body are suggestive but not specific to the infection. The other forms were not conclusive of the presence of HP.

RACIONAL: Estudos com tecnologias mais recentes como endoscopia com magnificação e cromoscopia mostraram que vários aspectos endoscópicos estão claramente associados à infecção por Helicobacter pylori. A descrição de padrões diferenciados de enantema no corpo gástrico através da magnificação de imagens reavivou o interesse na identificação desses padrões pela endoscopia convencional. OBJETIVO: Validar os padrões morfológicos de mucosa gástrica usando videogastroendoscopia convencional relacionados à gastrite por infecção por Helicobacter pylori, permitindo previsibilidade do seu diagnóstico e o direcionamento de biópsias. MÉTODOS: Estudo prospectivo de 339 pacientes consecutivos com análise das imagens de videogastroendoscopia obtidas, gravadas e armazenadas em banco de dados. Estas imagens foram estudadas com relação à presença ou não do Helicobacter pylori diagnosticado por teste rápido de urease e/ou por pesquisa direta por estudo anatomopatológico. Foram estudados: a) aspecto normal da mucosa; b) nodularidade da mucosa; c) enantema inespecífico difuso de antro e corpo; d) enantema em mosaico ou salpicado; e) enantema em estrias ou faixas; f) erosões elevadas; g) erosões planas; h) pólipos de glândulas fúndicas. Os principais critérios de exclusão foram o uso de medicamentos, tratamento prévio de HP e outras entidades que pudessem interferir nos resultados. RESULTADOS: Aplicando os critérios de exclusão, incluíram-se 170 dos 339 pacientes sendo 52 (30,58%) positivos para Helicobacter pylori e 118 negativos. No grupo positivo os achados que mais se associaram com a infecção foram: nodularidade no antro (26,92%); presença de erosões elevadas (15,38%) e mucosa em mosaico no corpo (21,15%). No grupo negativo o aspecto normal da mucosa foi de 66,94%; enantema em estrias ou faixas em 9,32%; erosões planas em 11,86%; e pólipos de glândulas fúndicas 11,86%. CONCLUSÃO: Achados endoscópicos são úteis na previsibilidade de localização e direcionamento de biópsias na pesquisa do HP. A mais representativa forma de gastrite por HP foi o achado de nodularidade na mucosa antral. As erosões elevadas e mucosa em mosaico no corpo são sugestivas, mas não específicas da infecção. As demais formas não foram conclusivas da presença do HP.

Elucidating Protein Involvement in the Stabilization of the Biogenic Silver Nanoparticles.

Silver nanoparticles (AgNPs) have been broadly used as antibacterial and antiviral agents. Further, interests for green AgNP synthesis have increased in recent years and several results for AgNP biological synthesis have been reported using bacteria, fungi and plant extracts. The understanding of the role and nature of fungal proteins, their interaction with AgNPs and the subsequent stabilization of nanosilver is yet to be deeply investigated. Therefore, in an attempt to better understand biogenic AgNP stabilization with the extracellular fungal proteins and to describe these supramolecular interactions between proteins and silver nanoparticles, AgNPs, produced extracellularly by Aspergillus tubingensis-isolated as an endophytic fungus from Rizophora mangle-were characterized in order to study their physical characteristics, identify the involved proteins, and shed light into the interactions among protein-NPs by several techniques. AgNPs of around 35 nm in diameter as measured by TEM and a positive zeta potential of +8.48 mV were obtained. These AgNPs exhibited a surface plasmon resonance (SPR) band at 440 nm, indicating the nanoparticles formation, and another band at 280 nm, attributed to the electronic excitations in tryptophan, tyrosine, and/or phenylalanine residues in fungal proteins. Fungal proteins were covalently bounded to the AgNPs, mainly through S-Ag bonds due to cysteine residues (HS-) and with few N-Ag bonds from H2N- groups, as verified by Raman spectroscopy. Observed supramolecular interactions also occur by electrostatic and other protein-protein interactions. Furthermore, proteins that remain free on AgNP surface may perform hydrogen bonds with other proteins or water increasing thus the capping layer around the AgNPs and consequently expanding the hydrodynamic diameter of the particles (~264 nm, measured by DLS). FTIR results enabled us to state that proteins adsorbed to the AgNPs did not suffer relevant secondary structure alteration upon their physical interaction with the AgNPs or when covalently bonded to them. Eight proteins in the AgNP dispersion were identified by mass spectrometry analyses. All these proteins are involved in metabolic pathways of the fungus and are important for carbon, phosphorous and nitrogen uptake, and for the fungal growth. Thereby, important proteins for fungi are also involved in the formation and stabilization of the biogenic AgNPs.

Conventional videoendoscopy can identify helicobacter pylori gastritis? / A videoendoscopia convencional pode identificar as gastrites por helicobacter pylori?

ABSTRACT Background: Studies with latest technologies such as endoscopy with magnification and chromoendoscopy showed that various endoscopic aspects are clearly related to infection by Helicobacter pylori (HP). The description of different patterns of erythema in gastric body under magnification of images revived interest in identifying these patterns by standard endoscopy. Aim: To validate the morphologic features of gastric mucosa related to H. pylori infection gastritis allowing predictability of their diagnosis as well as proper targeting biopsies. Methods: Prospective study of 339 consecutive patients with the standard videoendoscope image analysis were obtained, recorded and stored in a program database. These images were studied with respect to the presence or absence of H. pylori, diagnosed by rapid urease test and/or by histological analysis. Were studied: a) normal mucosa appearance; b) mucosal nodularity; c) diffuse nonspecific erythema or redness (with or without edema of folds and exudate) of antrum and body; d) mosaic pattern with focal area of hyperemia; e) erythema in streaks or bands (red streak); f) elevated (raised) erosion; g) flat erosions; h) fundic gland polyps. The main exclusion criteria were the use of drugs, HP pre-treatment and other entities that could affect results. Results: Applying the exclusion criteria, were included 170 of the 339 patients, of which 52 (30.58%) were positive for HP and 118 negative. On the positive findings, the most associated with infection were: nodularity in the antrum (26.92%); presence of raised erosion (15.38%) and mosaic mucosa in the body (21.15%). On the negative group the normal appearance of the mucosa was 66.94%; erythema in streaks or bands in 9.32%; flat erosions 11.86%; and fundic gland polyps 11.86%. Conclusion: Endoscopic findings are useful in the predictability of the result and in directing biopsies. The most representative form of HP related gastritis was the nodularity of the antral mucosa. The raised erosion and mucosa in mosaic in the body are suggestive but not specific to the infection. The other forms were not conclusive of the presence of HP.

RESUMO Racional: Estudos com tecnologias mais recentes como endoscopia com magnificação e cromoscopia mostraram que vários aspectos endoscópicos estão claramente associados à infecção por Helicobacter pylori. A descrição de padrões diferenciados de enantema no corpo gástrico através da magnificação de imagens reavivou o interesse na identificação desses padrões pela endoscopia convencional. Objetivo: Validar os padrões morfológicos de mucosa gástrica usando videogastroendoscopia convencional relacionados à gastrite por infecção por Helicobacter pylori, permitindo previsibilidade do seu diagnóstico e o direcionamento de biópsias. Métodos: Estudo prospectivo de 339 pacientes consecutivos com análise das imagens de videogastroendoscopia obtidas, gravadas e armazenadas em banco de dados. Estas imagens foram estudadas com relação à presença ou não do Helicobacter pylori diagnosticado por teste rápido de urease e/ou por pesquisa direta por estudo anatomopatológico. Foram estudados: a) aspecto normal da mucosa; b) nodularidade da mucosa; c) enantema inespecífico difuso de antro e corpo; d) enantema em mosaico ou salpicado; e) enantema em estrias ou faixas; f) erosões elevadas; g) erosões planas; h) pólipos de glândulas fúndicas. Os principais critérios de exclusão foram o uso de medicamentos, tratamento prévio de HP e outras entidades que pudessem interferir nos resultados. Resultados: Aplicando os critérios de exclusão, incluíram-se 170 dos 339 pacientes sendo 52 (30,58%) positivos para Helicobacter pylori e 118 negativos. No grupo positivo os achados que mais se associaram com a infecção foram: nodularidade no antro (26,92%); presença de erosões elevadas (15,38%) e mucosa em mosaico no corpo (21,15%). No grupo negativo o aspecto normal da mucosa foi de 66,94%; enantema em estrias ou faixas em 9,32%; erosões planas em 11,86%; e pólipos de glândulas fúndicas 11,86%. Conclusão: Achados endoscópicos são úteis na previsibilidade de localização e direcionamento de biópsias na pesquisa do HP. A mais representativa forma de gastrite por HP foi o achado de nodularidade na mucosa antral. As erosões elevadas e mucosa em mosaico no corpo são sugestivas, mas não específicas da infecção. As demais formas não foram conclusivas da presença do HP.

Quantification of trace O-containing compounds in GTL process samples via Fischer-Tropsch reaction by comprehensive two-dimensional gas chromatography/mass spectrometry.

Comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC-TOFMS) was successfully applied to eight real Brazilian Fischer-Tropsch (FT) product samples for the quantitative analysis of O-containing compounds. It not only allowed identifying and quantifying simultaneously a large number of O-containing compounds but also resolved many co-eluting components, such as carboxylic acids, which co-elute in one-dimensional gas chromatography. The homologous series of alcohols and carboxylic acids as trimethylsilyl derivatives were detected and identified at trace levels. The absolute quantification of each compound was accomplished with reliability using analytical curves. Linear alcohols (from C5 to C19), branched alcohols (C6-C13) and carboxylic acids (C4 to C12) were obtained in the range of 1.58 mg g(-1) to 14.75 mg g(-1), 0.51 mg g(-1) to 1.12 mg g(-1) and 0.21 mg g(-1) to 1.63 mg g(-1) of FT product samples, respectively. GC×GC-TOFMS provided a linear range (from 0.3 ng µL(-1) to 10 ng µL(-1)), good precision (<8%), and excellent accuracy (recovery range of 77% to 118%) for quantification of individual O-containing compounds in FT product samples. The results can benefit the development of gas-to-liquid technologies from natural gas and guide the choice of an FT conversion process that generates clean products with higher added value.

Qualidade de vida de pacientes idosos vivendo com HIV/AIDS / Quality of life in elderly patients living with HIV/AIDS / La calidad de vida de los pacientes de edad avanzada que viven con el VIH/SIDA

O objetivo foi avaliar a qualidade de vida de idosos com HIV/AIDS e correlacioná-la às características sociodemográficas, econômicas e clínicas. Estudo transversal analítico que avaliou idosos que vivem com HIV/AIDS. Coletadas informações sociodemográficas, mórbidas e situação econômica de acordo com o Critério de Classificação Econômica Brasil, e Targeted Quality of Life Instrument (HAT-QoL) que avalia qualidade de vida. Participaram 201 idosos, do sexo masculino (63,7%), solteiros ou divorciados (51,3%), classe econômica C, D ou E (61,7%) e baixa escolaridade (53,7%). A forma de contágio prevalente foi a sexual 130 (64,7%). Apresentaram correlação significativa com dois ou até sete domínios do HAT-QoL: idade, renda individual e familiar, e tempo de diagnóstico. A qualidade de vida dos idosos que vivem com HIV/AIDS apresentou menores escores nos domínios preocupação com o sigilo, atividade sexual e preocupação financeira. Idade, renda e tempo de diagnóstico foram as variáveis que mais responderam pela alteração da qualidade de vida.

This study’s objective was to evaluate quality of life in older adults living with HIV/AIDS and the associations with socio-demographic, economic, and clinical characteristics, using a cross-sectional design. Data were collected on demographics, disease history, and economic status according to the Brazilian Economic Classification Criterion and Targeted Quality of Life Instrument (HAT-QoL). A total of 201 elderly participated, with the following characteristics: males (63.7%), single or divorced (51.3%), economic classes C, D, or E (61.7%), and low schooling (53.7%). Nearly two-thirds (130, or 64.7%) had been infected by the sexual route. The following were significantly correlated with two or up to seven domains in the HAT-QoL: age, individual and family income, and time since diagnosis. Quality of life in elderly living with HIV/AIDS showed the lowest scores in the domains related to concern over disclosure of their diagnosis, sexual activity, and financial concerns. Age, income, and time since diagnosis were the variables that most accounted for altered quality of life.

El objetivo fue evaluar calidad de vida de adultos mayores con VIH/SIDA y correlacionarlos con sus características sociodemográficas, económicas y clínicas. Se trata de un estudio transversal analítico que evaluó a pacientes ancianos, que viven con VIH/SIDA. Se analizan factores sociodemográficos, económicos y morbilidad, de acuerdo con el Criterio de Clasificación Económica de Brasil y el Targeted Quality of Life Instrument (HAT-QoL) para evaluar la calidad de vida. Los participantes fueron 201 hombres de edad avanzada (63,7%), solteros o divorciados (51,3%), con perfil económico clase C, D o E (61,7%) y baja escolaridad (53,7%). La forma predominante de transmisión fue sexual 130 (64,7%) y la comorbilidad más común cardiovascular 69 (34,3%). Las variables que se correlacionaron significativamente con siete dominios de HAT-QOL fueron: edad, renta personal y familiar y tiempo transcurrido desde el diagnóstico. La calidad de vida de los pacientes se ha visto comprometida en algunos aspectos: preocupaciones sobre confidencialidad, actividad sexual y preocupación financiera. Edad, ingresos y tiempo transcurrido desde el diagnóstico fueron variables que explicaron un cambio de calidad de vida.

Imidate-based cross-linkers for structural proteomics: increased charge of protein and peptide ions and CID and ECD fragmentation studies.

Chemical cross-linking is an attractive low-resolution technique for structural studies of protein complexes. Distance constraints obtained from cross-linked peptides identified by mass spectrometry (MS) are used to construct and validate protein models. Amidinating cross-linkers such as diethyl suberthioimidate (DEST) have been used successfully in chemical cross-linking experiments. In this work, the application of a commercial diimidate cross-linking reagent, dimethyl suberimidate (DMS), was evaluated with model peptides and proteins. The peptides were designed with acetylated N-termini followed by random sequences containing two Lys residues separated by an Arg residue. After cross-linking reactions, intra- and intermolecular cross-linked species were submitted to CID and ECD dissociations to study their fragmentation features in the gas phase. Fragmentation of intramolecular peptides by collision induced dissociation (CID) demonstrates a unique two-step fragmentation pathway involving formation of a ketimine as intermediate. Electron capture and electron transfer dissociation (ECD and ETD) experiments demonstrated that the cyclic moiety is not dissociated. Intermolecular species demonstrated previously described fragmentation behavior in both CID and ECD experiments. The charge state distributions (CSD) obtained after reaction with DMS were compared with those obtained with disuccinimidyl suberate (DSS). CSDs for peptides and proteins were increased after their reaction with DMS, owing to the higher basicity of DMS modified species. These features were also observed in LC-MS experiments with bovine carbonic anhydrase II (BCA) after cross-linking with DMS and tryptic proteolysis. Cross-linked peptides derived from this protein were identified at high confidence and those species were in agreement with the crystal structure of BCA.

A new platinum complex with tryptophan: synthesis, structural characterization, DFT studies and biological assays in vitro over human tumorigenic cells.

A new platinum(II) complex with the amino acid L-tryptophan (trp), named Pt-trp, was synthesized and characterized. Elemental, thermogravimetric and ESI-QTOF mass spectrometric analyses led to the composition [Pt(C11H11N2O2)2]⋅6H2O. Infrared spectroscopic data indicate the coordination of trp to Pt(II) through the oxygen of the carboxylate group and also through the nitrogen atom of the amino group. The (13)C CP/MAS NMR spectroscopic data confirm coordination through the oxygen atom of the carboxylate group, while the (15)N CP/MAS NMR data confirm coordination of the nitrogen of the NH2 group to the metal. Density functional theory (DFT) studies were applied to evaluate the cis and trans coordination modes of trp to platinum(II). The trans isomer was shown to be energetically more stable than the cis one. The Pt-trp complex was evaluated as a cytotoxic agent against SK-Mel 103 (human melanoma) and Panc-1 (human pancreatic carcinoma) cell lines. The complex was shown to be cytotoxic over the considered cells.

The Biginelli reaction with an imidazolium-tagged recyclable iron catalyst: kinetics, mechanism, and antitumoral activity.

The present work describes the synthesis, characterization, and application of a new ion-tagged iron catalyst. The catalyst was employed in the Biginelli reaction with impressive performance. High yields have been achieved when the reaction was carried out in imidazolium-based ionic liquids (BMI⋅PF6, BMI⋅NTf2, and BMI⋅BF4), thus showing that the ionic-liquid effects play a role in the reaction. Moreover, the ion-tagged catalyst could be recovered and reused up to eight times without any noticeable loss in activity. Mechanistic studies performed by using high-resolution electrospray-ionization quadrupole-time-of-flight mass (HR-EI-QTOF) spectrometry and kinetic experiments indicate only one reaction pathway and rule out the other two possibilities under the development conditions. The theoretical calculations are in accordance with the proposed mechanism of action of the iron catalyst. Finally, the 37 dihydropyrimidinone derivatives, products of the Biginelli reaction, had their cytotoxicity evaluated in assays against MCF-7 cancer cell linages with encouraging results of some derivatives, which were virtually non-toxic against healthy cell linages (fibroblasts).