One-dimensional convolutional neural network model for breast cancer subtypes classification and biochemical content evaluation using micro-FTIR hyperspectral images

Breast cancer treatment still remains a challenge, where molecular subtypes classification plays a crucial role in selecting appropriate and specific therapy. The four subtypes are Luminal A (LA), Luminal B (LB), HER2 subtype, and Triple-Negative Breast Cancer (TNBC). Immunohistochemistry is the gold-standard evaluation, although interobserver variations are reported and molecular signatures identification is time-consuming. Fourier transform infrared micro-spectroscopy with machine learning approaches have been used to evaluate cancer samples, presenting biochemical-related explainability. However, this explainability is harder when using deep learning. This study created a 1D deep learning tool for breast cancer subtype evaluation and biochemical contribution. Sixty hyperspectral images were acquired from a human breast cancer microarray. K-Means clustering was applied to select tissue and paraffin spectra. CaReNet-V1, a novel 1D convolutional neural network, was developed to classify breast cancer (CA) and adjacent tissue (AT), and molecular subtypes. A 1D adaptation of Grad-CAM was applied to assess the biochemical impact to the classifications. CaReNet-V1 effectively classified CA and AT (test accuracy of 0.89), as well as HER2 and TNBC subtypes (0.83 and 0.86), with greater difficulty for LA and LB (0.74 and 0.68). The model enabled the evaluation of the most contributing wavenumbers to the predictions, providing a direct relationship with the biochemical content. Therefore, CaReNet-V1 and hyperspectral images is a potential approach for breast cancer biopsies assessment, providing additional information to the pathology report. Biochemical content impact feature may be used for other studies, such as treatment efficacy evaluation and development new diagnostics and therapeutic methods.Comment: 23 pages, 5 figures, 2 table

Dual-path convolutional neural network using micro-FTIR imaging to predict breast cancer subtypes and biomarkers levels: estrogen receptor, progesterone receptor, HER2 and Ki67

Breast cancer molecular subtypes classification plays an import role to sort patients with divergent prognosis. The biomarkers used are Estrogen Receptor (ER), Progesterone Receptor (PR), HER2, and Ki67. Based on these biomarkers expression levels, subtypes are classified as Luminal A (LA), Luminal B (LB), HER2 subtype, and Triple-Negative Breast Cancer (TNBC). Immunohistochemistry is used to classify subtypes, although interlaboratory and interobserver variations can affect its accuracy, besides being a time-consuming technique. The Fourier transform infrared micro-spectroscopy may be coupled with deep learning for cancer evaluation, where there is still a lack of studies for subtypes and biomarker levels prediction. This study presents a novel 2D deep learning approach to achieve these predictions. Sixty micro-FTIR images of 320x320 pixels were collected from a human breast biopsies microarray. Data were clustered by K-means, preprocessed and 32x32 patches were generated using a fully automated approach. CaReNet-V2, a novel convolutional neural network, was developed to classify breast cancer (CA) vs adjacent tissue (AT) and molecular subtypes, and to predict biomarkers level. The clustering method enabled to remove non-tissue pixels. Test accuracies for CA vs AT and subtype were above 0.84. The model enabled the prediction of ER, PR, and HER2 levels, where borderline values showed lower performance (minimum accuracy of 0.54). Ki67 percentage regression demonstrated a mean error of 3.6%. Thus, CaReNet-V2 is a potential technique for breast cancer biopsies evaluation, standing out as a screening analysis technique and helping to prioritize patients.Comment: 32 pages, 3 figures, 6 table

Selection of aptamers against the Jagged-1 protein

The breast cancer is one of the biggest public health problems in the world, being the main female type of cancer that affects the population. The Jagged-1 protein plays an important role in the biology and development of cancer, influencing angiogenesis, growth of neoplastic cells, tumor stem cells, epithelial mesenchymal transition, metastatic processes and resistance to therapies in various types of cancer. In this project, our aim was to select an aptamer for JAG1 ligand using an aptamer library, that could be used as a radiopharmaceutical for PET / SPECT / CT diagnosis of tumors that express JAG1.  Our work showed that MDA-MB-231-JAG1 cells overexpress more mRNA and JAG1 protein than control cells (MDA-MB-231-Control). We also selected aptamers with high affinity for MDA-MB-231-JAG1 cells that could be a useful tool for the development of new radiopharmaceuticals for the diagnosis and treatment of tumors that overexpress JAG1.  

UM ESTUDO DA UTILIZAÇÃO DA CONTROLADORIA NA GESTÃO HOSPITALAR

The Controllership can support managers in their decision-making, seeking the future improvement of organizations. Based on this explanation, a research was carried out with the objective of verifying that the Controllership has been used as a decision-making tool in the management of Brazilian hospitals and the benefits derived from its use for these organizations. For the study, a qualitative, exploratory-descriptive and bibliographic research was carried out. The bibliographic research was carried out by means of the survey of the articles published in Portuguese in the Scholar Google database from 2012 to 2018. The results found have shown that, in general, hospitals make little use of the instruments provided by the Controllership. Thus, it was possible to confirm the hypothesis of the work of which hospital management makes use of the Controllership in its control activities, but does not use it in its decision making.A Controladoria pode embasar os gestores em suas tomadas de decisões, buscando a melhoria futura das organizações. Partindo desta explanação, foi realizada uma pesquisa com o objetivo de verificar se a Controladoria tem sido utilizada como ferramenta de tomada de decisão na gestão dos hospitais brasileiros e quais os benefícios advindos de sua utilização para essas organizações. Para o estudo foi efetuada uma pesquisa dos tipos qualitativa, exploratório-descritiva e bibliográfica. A pesquisa bibliográfica foi realizada por meio do levantamento dos artigos publicados em português na base de dados Google Acadêmico no período de 2012 a 2018. Os resultados encontrados demonstraram que de maneira geral, os hospitais fazem pouco uso dos instrumentos disponibilizados pela Controladoria. Assim, foi possível confirmar a hipótese do trabalho de que gestão dos hospitais faz uso da Controladoria em suas atividades de controle, porém não a utilizam em suas tomadas de decisões

Evidence for glycosylation on a DNA-binding protein of Salmonella enterica

BACKGROUND: All organisms living under aerobic atmosphere have powerful mechanisms that confer their macromolecules protection against oxygen reactive species. Microorganisms have developed biomolecule-protecting systems in response to starvation and/or oxidative stress, such as DNA biocrystallization with Dps (DNA-binding protein from starved cells). Dps is a protein that is produced in large amounts when the bacterial cell faces harm, which results in DNA protection. In this work, we evaluated the glycosylation in the Dps extracted from Salmonella enterica serovar Typhimurium. This Dps was purified from the crude extract as an 18-kDa protein, by means of affinity chromatography on an immobilized jacalin column. RESULTS: The N-terminal sequencing of the jacalin-bound protein revealed 100% identity with the Dps of S. enterica serovar Typhimurium. Methyl-alpha-galactopyranoside inhibited the binding of Dps to jacalin in an enzyme-linked lectin assay, suggesting that the carbohydrate recognition domain (CRD) of jacalin is involved in the interaction with Dps. Furthermore, monosaccharide compositional analysis showed that Dps contained mannose, glucose, and an unknown sugar residue. Finally, jacalin-binding Dps was detected in larger amounts during the bacterial earlier growth periods, whereas high detection of total Dps was verified throughout the bacterial growth period. CONCLUSION: Taken together, these results indicate that Dps undergoes post-translational modifications in the pre- and early stationary phases of bacterial growth. There is also evidence that a small mannose-containing oligosaccharide is linked to this bacterial protein

Characterization of the mechanisms underlying the crosstalk between galectins and notch in gastric cancer

Gastric cancer is the fourth most common cancer and the second leading cause of cancer-related deaths worldwide. Galectins form a family of β-galactosides binding proteins that recognize a variety of glycan-containing proteins at the cell surface and are overexpressed in various tumors, including gastric cancer. Galectins overexpression as well as changes in their subcellular distribution has been associated with gastric cancer progression and poor prognosis. It is not well understood, however, how the interaction between galectins and glycosylated receptors modulates tumor development and growth. Since Notch receptors and ligands contain glycan structures known to bind galectins, we aim to demonstrate that galectins expression in the tumor microenvironment may interfere with Notch signaling activation during tumor development and progression.\ud \ud Materials and methods\ud Immunoprecipitation procedures with gastric cancer cell line AGS (ATCC CRL-1739) and MKN45 (ACC 409) were used to test for association between galectin-1/-3 and Notch-1 receptor. Furthermore, we transfected AGS cell line with siRNA against galectin-1/-3 or scramble using standard protocols (IDT DNA technologies), stimulate them with immobilized human recombinant delta-4 or Jagged-1 and assessed Notch-1 receptor activation.\ud \ud Results\ud Galectin-1 and -3 interact with Notch-1 receptor and differentially modulate Notch signaling pathway upon activation by Delta/Jagged ligands. Galectin-1 knockdown alters Notch-1 activation induced by Delta-4 whereas galectin-3 knockdown alters jagged-1-mediated Notch-1 activation. Furthermore, we found that exogenously added galectin-3 can enhance Notch-1 activation by Jagged-1.\ud \ud Conclusion\ud Our results suggest that galectin-1 and -3 interact with Notch-1 receptor and differentially modulate Notch signaling activation induced by Jagged-1 and Delta-4

Losartán y sus derivados como radiotrazadores del receptor tipo 1 de agiotensina II

El receptor tipo 1 de angiotensina II (AT1R) juega un papel fundamental en las enfermedades cardiovasculares y se ha reconocido, en los últimos diez años, por su activa expresión en varios tipos de células y tejidos tumorales propiciando la progresión del cáncer. Los fármacos antagonistas de este receptor, como el losartán, utilizados para el control de la hipertensión arterial han demostrado un potencial efecto antitumoral al ser capaces de reducir la progresión del tumor, la vascularización y la metastásis. El objetivo de este trabajo es brindar una panorámica sobre los estudios más recientes de los derivados marcados del losartán, destacando su posible utilización como radiotrazadores del AT1R en tumores que lo expresen. Derivados del losartán marcados con tecnecio-99 metaestable, carbono-11 y fluor-18 permiten detectar y cuantificar el AT1Ren algunos órganos como el corazón y los riñones. Se demuestra la relación existente entre el losartán y el cáncer según los resultados de estudios preclínicos, se resumen diferentes vías de obtención de sus derivados marcados y los resultados de su evaluación como radiotrazadores biológicos del AT1R. En las conclusiones se señala que los derivados de losartán marcados con 99mTc, 11C y 18F pudieran ser utilizados como radiotrazadores para detectar y cuantificar el AT1R en células y tejidos tumorales de pacientes utilizando la imagen molecular como técnica no invasiva.  Existe un amplio campo de investigación relacionado con el desarrollo de nuevos radiotrazadores SPECT o PET del AT1R con fines diagnósticos tanto en patologías como la hipertensión y enfermedades cardiovasculares, como en oncología

Eficácia do treinamento da musculatura do assoalho pélvico e de exercícios hipopressivos para o tratamento do prolapso de órgãos pélvicos em mulheres: ensaio clínico randomizado

CONTEXT AND OBJECTIVE: Previous studies have shown that women with pelvic floor dysfunctions present decreased cross-sectional area (CSA) of the levator ani muscle. One way to assess the effects of training programs is to measure the CSA of the muscle, using ultrasonography. The aim here was to evaluate the efficacy of pelvic floor muscle training and hypopressive exercises for increasing the CSA of the levator ani muscle in women with pelvic organ prolapse. DESIGN AND SETTING: Prospective randomized controlled trial at the Urogynecology outpatient clinic of Universidade Federal de São Paulo (UNIFESP). METHODS: Fifty-eight women with stage II pelvic organ prolapse were divided into three groups for physiotherapy: a pelvic floor muscle training group (GI); a hypopressive exercise group (GII); and a control group (GIII). The patients underwent transperineal ultrasonographic evaluation using a transducer of frequency 4-9 MHz. The (CSA) of the levator ani muscle was measured before physiotherapy and after 12 weeks of treatment. RESULTS: The groups were homogeneous regarding age, number of pregnancies, number of vaginal deliveries, body mass index and hormonal status. Statistically significant differences in CSA were found in GI and GII from before to after the treatment (P < 0.001), but not in relation to GIII (P = 0.816). CONCLUSIONS: The CSA of the levator ani muscle increased significantly with physiotherapy among the women with pelvic organ prolapse. Pelvic floor muscle training and hypopressive exercises produced similar improvements in the CSA of the levator ani muscle.CONTEXTO E OBJETIVO: Estudos anteriores mostraram que mulheres com disfunção do assoalho pélvico possuem diminuição da área de secção transversal (AST) do músculo levantador do ânus. Uma forma de avaliar os efeitos de um programa de treinamento é mensurar a AST do músculo por ultrassonografia. O objetivo foi avaliar a eficácia do treinamento da musculatura do assoalho pélvico e de exercícios hipopressivos no aumento da AST do músculo levantador do ânus em mulheres com prolapso de órgãos pélvicos. TIPO DE ESTUDO E LOCAL: Estudo prospectivo, randomizado e controlado realizado no Ambulatório de Uroginecologia da Universidade Federal de São Paulo (UNIFESP). MÉTODOS: Cinquenta e oito mulheres com prolapso de órgãos pélvicos estádio II foram divididas em três grupos para tratamento fisioterapêutico: (GI) grupo de treinamento dos músculos do assoalho pélvico, (GII) grupo de exercícios hipopressivos e (GIII) grupo controle. As pacientes se submeteram a avaliação ultrassonográfica transperineal com transdutor de frequência 4-9 MHz. Foi mensurada a AST do músculo levantador do ânus antes e após 12 semanas de tratamento fisioterapêutico. RESULTADOS: Os grupos foram homogêneos no que se refere a idade, número de gestações, número de partos vaginais, índice de massa corpórea e estado hormonal. Diferença significante foi observada na AST de GI e GII antes e após o tratamento (P < 0,001) e isso não ocorreu com o GIII (P = 0,816). CONCLUSÕES: A AST do músculo levantador do ânus aumentou significativamente com tratamento fisioterapêutico em mulheres com prolapso de órgãos pélvicos. Treinamento dos músculos do assoalho pélvico e exercícios hipopressivos são semelhantes no que se refere ao aumento da AST do músculo levantador do ânus.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo (UNIFESP) Department of GynecologyUniversidade Federal de Uberlândia Department of Gynecology and ObstetricsUNIFESP, Department of GynecologyFAPESP: 2007/08246-8SciEL

Galectin-3 negatively regulates the frequency and function of CD4+CD25+Foxp3+ regulatory T cells and influences the course of Leishmania major infection

Galectin-3, an endogenous glycan-binding protein, plays essential roles during microbial infection by modulating innate and adaptive immunity. However, the role of galectin-3 within the CD4+CD25+Foxp3+ T regulatory (TREG) cell compartment has not yet been explored. Here, we found, in a model of Leishmania major infection, that galectin-3 deficiency increases the frequency of peripheral TREG cells both in draining lymph nodes (LNs) and sites of infection. These observations correlated with an increased severity of the disease, as shown by increased footpad swelling and parasite burden. Galectin-3-deficient (Lgals3−/−) TREG cells displayed higher CD103 expression, showed greater suppressive capacity, and synthesized higher amounts of IL-10 compared with their wild-type (WT) counterpart. Furthermore, both TREG cells and T effector (TEFF) cells from Lgals3−/− mice showed higher expression of Notch1 and the Notch target gene Hes-1. Interestingly, Notch signaling components were also altered in both TREG and TEFF cells from uninfected Lgals3−/− mice. Thus, endogenous galectin-3 regulates the frequency and function of CD4+CD25+Foxp3+ TREG cells and alters the course of L. major infection.Fil: Fermino, Marise L.. Universidade de Sao Paulo; BrasilFil: Dias, Fabrício C.. Universidade de Sao Paulo; BrasilFil: Lopes, Carla D.. Universidade de Sao Paulo; BrasilFil: Souza, Maria A.. Universidade de Sao Paulo; BrasilFil: Cruz, Ângela K.. Universidade de Sao Paulo; BrasilFil: Liu, Fu Tong. University of California at Davis; Estados UnidosFil: Chammas, Roger. Universidade de Sao Paulo; BrasilFil: Roque Barreira, Maria C.. Universidade de Sao Paulo; BrasilFil: Rabinovich, Gabriel Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Bernardes, Emerson S.. Universidade de Sao Paulo; Brasi

Synthesis and Evaluation of [F-18]FEtLos and [F-18]AMBF(3)Los as Novel F-18-Labelled Losartan Derivatives for Molecular Imaging of Angiotensin II Type 1 Receptors

Losartan is widely used in clinics to treat cardiovascular related diseases by selectively blocking the angiotensin II type 1 receptors (AT(1)Rs), which regulate the renin-angiotensin system (RAS). Therefore, monitoring the physiological and pathological biodistribution of AT(1)R using positron emission tomography (PET) might be a valuable tool to assess the functionality of RAS. Herein, we describe the synthesis and characterization of two novel losartan derivatives PET tracers, [F-18]fluoroethyl-losartan ([F-18]FEtLos) and [F-18]ammoniomethyltrifluoroborate-losartan ([F-18]AMBF(3)Los). [F-18]FEtLos was radiolabeled by F-18-fluoroalkylation of losartan potassium using the prosthetic group 2-[F-18]fluoroethyl tosylate; whereas [F-18]AMBF(3)Los was prepared following an one-step F-18-F-19 isotopic exchange reaction, in an overall yield of 2.7 +/- 0.9% and 11 +/- 4%, respectively, with high radiochemical purity (>95%). Binding competition assays in AT(1)R-expressing membranes showed that AMBF(3)Los presented an almost equivalent binding affinity (K-i 7.9 nM) as the cold reference Losartan (K-i 1.5 nM), unlike FEtLos (K-i 2000 nM). In vitro and in vivo assays showed that [F-18]AMBF(3)Los displayed a good binding affinity for AT(1)R-overexpressing CHO cells and was able to specifically bind to renal AT(1)R. Hence, our data demonstrate [F-18]AMBF(3)Los as a new tool for PET imaging of AT(1)R with possible applications for the diagnosis of cardiovascular, inflammatory and cancer diseases