Development of a novel five dye insertion/deletion (INDEL) panel for ancestry determination.

The use of genetic markers, specifically Short Tandem Repeats (STRs), has been a valuable tool for identifying persons of interest. However, the ability to analyze additional markers including Single Nucleotide Polymorphisms (SNPs) and Insertion/Deletion (INDELs) polymorphisms allows laboratories to explore other investigative leads. INDELs were chosen in this study because large panels can be differentiated by size, allowing them to be genotyped by capillary electrophoresis. Moreover, these markers do not produce stutter and are smaller in size than STRs, facilitating the recovery of genetic information from degraded samples. The INDEL Ancestry Informative Markers (AIMs) in this study were selected from the 1000 Genomes Project based on a fixation index (FST) greater than 0.50, high allele frequency divergence, and genetic distance. A total of 25 INDEL-AIMs were optimized and validated according to SWGDAM guidelines in a five-dye multiplex. To validate the panel, genotyping was performed on 155 unrelated individuals from four ancestral groups (Caucasian, African, Hispanic, and East Asian). Bayesian clustering and principal component analysis (PCA) were performed revealing clear separation among three groups, with some observed overlap within the Hispanic group. Additionally, the PCA results were compared against a training set of 793 samples from the 1000 Genomes Project, demonstrating consistent results. Validation studies showed the assay to be reproducible, tolerant to common inhibitors, robust with challenging casework type samples, and sensitive down to 125 pg. In conclusion, our results demonstrated the robustness and effectiveness of a 25 loci INDEL system for ancestry inference of four ancestries commonly found in the United States.

Development of a novel five-dye panel for human identification insertion/deletion (INDEL) polymorphisms.

DNA analysis of forensic case samples relies on short tandem repeats (STRs), a key component of the combined DNA index system (CODIS) used to identify individuals. However, limitations arise when dealing with challenging samples, prompting the exploration of alternative markers such as single nucleotide polymorphisms (SNPs) and insertion/deletion (INDELs) polymorphisms. Unlike SNPs, INDELs can be differentiated easily by size, making them compatible with electrophoresis methods. It is possible to design small INDEL amplicons (<200 bp) to enhance recovery from degraded samples. To this end, a set of INDEL Human Identification Markers (HID) was curated from the 1000 Genomes Project, employing criteria including a fixation index (FST) ≤ 0.06, minor allele frequency (MAF) >0.2, and high allele frequency divergence. A panel of 33 INDEL-HIDs was optimized and validated following the Scientific Working Group on DNA Analysis Methods (SWGDAM) guidelines, utilizing a five-dye multiplex electrophoresis system. A small sample set (n = 79 unrelated individuals) was genotyped to assess the assay's performance. The validation studies exhibited reproducibility, inhibition tolerance, ability to detect a two-person mixture from a 4:1 to 1:6 ratio, robustness with challenging samples, and sensitivity down to 125 pg of DNA. In summary, the 33-loci INDEL-HID panel exhibited robust recovery with low-template and degraded samples and proved effective for individualization within a small sample set.

Application of time domain nuclear magnetic resonance (TD-NMR) for study of the distillation curve of petroleum.

Crude oil distillates are a highly useful industrial product, mainly for energy generation. Unfortunately, they are rarely studied, mainly due to the low accessibility to products directly obtained from the distillation process, which is a laborious, expensive, and time-consuming operation. This work presents and discusses the use of time-domain nuclear magnetic resonance (TD-NMR) as a simple, affordable, and straightforward tool for the development of correlations supported on the transverse relaxation time (T2 ) and boiling temperature. The results point out a high convergence between TD-NMR experimental data and the ASTM D2892 method for distillates from light, medium, and heavy oils, with up to 52.20% of accumulated mass and boiling point temperature (Tb ) up to 400°C. Furthermore, an unprecedented relationship between T2 values and the accumulated mass of the distillates is first demonstrated. This new insight opens new perspectives for future prediction of accumulated mass for unknown crude oils, placing the TD-NMR relaxometry as an appeal spectroscopy approach with a potential to meaningfully contribute to the daily refining petrochemical industry field operations.

Time domain NMR: An alternative for study of the asphaltenes precipitated in petroleum.

During the oil production and processing stages, the asphaltene precipitation is one of the great operation problems of oil industry. It can precipitate in the formation, tubing, or surface, causing operating problems, such as reduction in oil recovery by changing the reservoir permeability and wettability, clogging of the pipelines, and difficulty in separations process. The quantification of asphaltenes in petroleum by ASTM D6560 standard method is very laborious and use of a larger solvent volume than necessary. The present work proposes the use of time domain nuclear magnetic resonance (TD-NMR) as new methodology to quantify the asphaltene precipitated in crude oil. Three (light, medium, and heavy) crude oils with asphaltenes content of 0.97, 1.88, and 7.00 wt% were mixed with n-heptane in different R (ml of solvent/g of oil) values and analyzed by means of transverse relaxation time (T2 ). According NMR results, the R values enough for complete asphaltene precipitation for the oils A, B, and C were, respectively, equal to 16.50, 23.00, and 39.50 ml g-1 . These outcomes represent a reduction of 58.75%, 42.50%, and 1.25% in the solvent volume per mass of oil for the oil A, B, and C, respectively, compared to the ASTM D6560 method, which imposes 40 ml g-1 . Therefore, it has been shown that TD-NMR can be applied to estimate the amount of asphaltene precipitated in petroleum and have potential to be applied in routine analysis with advantages of saving time and costs.

IPOscore: An interactive web-based platform for postoperative surgical complications analysis and prediction in the oncology domain.

BACKGROUND: The performance of traditional risk score systems to predict (post)-operative outcomes is limited. This weakness reduces confidence in its use to support clinical risk mitigation decisions. However, the rapid growth of health data in the last years offers principles to deal with some of these limitations. In this regard, the data allows the extraction of relevant information for both patients stratification and the rigorous identification of associated risk factors. The patients can then be targeted to specific preoperative optimization programs, thus contributing to the reduction of associated morbidity and mortality. OBJECTIVES: The main goal of this work is, therefore, to provide a clinical decision support system (CDSS) based on data-driven modeling methods for surgical risk prediction specific for cancer patients in Portugal. RESULTS: The result is IPOscore, a single web-based platform aimed at being an innovative approach to assist clinical decision-making in the surgical oncology domain. This system includes a database to store/manage the clinical data collected in a structured format, data visualization and analysis tools, and predictive machine learning models to predict postoperative outcomes in cancer patients. IPOscore also includes a pattern mining module based on biclustering to assess the discriminative power of a pattern towards postsurgical outcomes. Additionally, a mobile application is provided to this end. CONCLUSIONS: The IPOscore platform is a valuable tool for surgical oncologists not only for clinical data management but also as a preventative and predictive healthcare system. Currently, this clinical support tool is being tested at the Portuguese Institute of Oncology (IPO-Porto), and can be accessed online at https://iposcore.org.

Pirfenidone Sensitizes NCI-H460 Non-Small Cell Lung Cancer Cells to Paclitaxel and to a Combination of Paclitaxel with Carboplatin.

Pirfenidone, an antifibrotic drug, has antitumor potential against different types of cancers. Our work explored whether pirfenidone sensitizes non-small cell lung cancer (NSCLC) cell lines to chemotherapeutic treatments. The cytotoxic effect of paclitaxel in combination with pirfenidone against three NSCLC cell lines (A549, NCI-H322 and NCI-H460) was evaluated using the sulforhodamine B assay. The effects of this combination on cell viability (trypan blue exclusion assay), proliferation (BrdU incorporation assay), cell cycle (flow cytometry following PI staining) and cell death (Annexin V-FITC detection assay and Western blot) were analyzed on the most sensitive cell line (NCI-H460). The cytotoxic effect of this drug combination was also evaluated against two non-tumorigenic cell lines (MCF-10A and MCF-12A). Finally, the ability of pirfenidone to sensitize NCI-H460 cells to a combination of paclitaxel plus carboplatin was assessed. The results demonstrated that pirfenidone sensitized NCI-H460 cells to paclitaxel treatment, reducing cell growth, viability and proliferation, inducing alterations in the cell cycle profile and causing an increase in the % of cell death. Remarkably, this combination did not increase cytotoxicity in non-tumorigenic cells. Importantly, pirfenidone also sensitized NCI-H460 cells to paclitaxel plus carboplatin. This work highlights the possibility of repurposing pirfenidone in combination with chemotherapy for the treatment of NSCLC.

Association Between Estrogen Receptors and GATA3 in Bladder Cancer: A Systematic Review and Meta-Analysis of Their Clinicopathological Significance.

Background: Estrogen receptors alpha (ERα) and beta (ERß) and the cooperating protein GATA-binding factor 3 (GATA3) have been implicated in bladder carcinogenesis and tumour progression. GATA3 and ER have been functionally linked in the establishment of luminal fate in breast tissue, but to date their relationship in bladder cancer has not been established. This information will be useful to advance diagnostic and prognostic markers. Aim: To determine the relationship between the expression of ERα, ERß and GATA3 in bladder cancer, disclose their prognostic and diagnostic value and their association with clinicopathological characteristics. Methods: A comprehensive literature search in PubMed database was performed for all immunohistochemical studies of ERα, ERß and/or GATA3 in bladder cancer patients. We selected eligible studies in accordance with the PRISMA guidelines and evaluated methodological quality and risk of bias based on quality criteria from the reporting recommendations for tumour MARKer (REMARK) prognostic studies. Risk of bias assessment was performed using Review Manager 5. R software was used for all statistical analysis, the packages used were meta and dmetar for the standard meta-analysis, and netmeta for the network meta-analysis. Results: Thirteen studies were eligible for ERα, 5 for ERß and 58 for GATA3 meta-analysis. Low grade tumours showed significantly lower ERα expression. GATA3 was widely expressed in bladder tumours, especially urothelial carcinomas, with higher expression of GATA3 in low grade and low stage tumours. Data was insufficient to determine the prognostic value of either ERα or ERß, but GATA3-positivity was associated with higher recurrence free survival. A negative correlation between ERα or ERß positivity and GATA3 expression was disclosed. Additionally, several sources of heterogeneity were identified, which can be used to improve future studies. Conclusion: The clinicopathological value of ERα and ERß was inconclusive due to low availability of studies using validated antibodies. Still, this meta-analysis supports GATA3 as good prognostic marker. On the contrary, ERα-positivity was associated to higher grade tumours; while ERα and ERß were inversely correlated with GATA3 expression. Considering that it has previously been shown that bladder cancer cell lines have functional ERs, this suggests that ERα could be activated in less differentiated cells and independently of GATA3. Therefore, a comprehensive analysis of ERα and ERß expression in BlaCa supported by complete patient clinical history is required for the identification of BlaCa subtypes and subgroups of patients expressing ERα, to investigate if they could benefit from treatment with hormonal therapy. Systematic Review Registration: Prospero, CRD42021226836.

ST6Gal1 targets the ectodomain of ErbB2 in a site-specific manner and regulates gastric cancer cell sensitivity to trastuzumab.

The clinical performance of the therapeutic monoclonal antibody trastuzumab in the treatment of ErbB2-positive unresectable gastric cancer (GC) is severely hampered by the emergence of molecular resistance. Trastuzumab's target epitope is localized within the extracellular domain of the oncogenic cell surface receptor tyrosine kinase (RTK) ErbB2, which is known to undergo extensive N-linked glycosylation. However, the site-specific glycan repertoire of ErbB2, as well as the detailed molecular mechanisms through which specific aberrant glycan signatures functionally impact the malignant features of ErbB2-addicted GC cells, including the acquisition of trastuzumab resistance, remain elusive. Here, we demonstrate that ErbB2 is modified with both α2,6- and α2,3-sialylated glycan structures in GC clinical specimens. In-depth mass spectrometry-based glycomic and glycoproteomic analysis of ErbB2's ectodomain disclosed a site-specific glycosylation profile in GC cells, in which the ST6Gal1 sialyltransferase specifically targets ErbB2 N-glycosylation sites occurring within the receptor's trastuzumab-binding domain. Abrogation of ST6Gal1 expression reshaped the cellular and ErbB2-specific glycomes, expanded the cellular half-life of the ErbB2 receptor, and sensitized ErbB2-dependent GC cells to trastuzumab-induced cytotoxicity through the stabilization of ErbB dimers at the cell membrane, and the decreased activation of both ErbB2 and EGFR RTKs. Overall, our data demonstrates that ST6Gal1-mediated aberrant α2,6-sialylation actively tunes the resistance of ErbB2-driven GC cells to trastuzumab.

Chitinase 3-like-1 and fibronectin in the cargo of extracellular vesicles shed by human macrophages influence pancreatic cancer cellular response to gemcitabine.

Tumour-associated macrophages have been implicated in pancreatic ductal adenocarcinoma (PDAC) therapy response and Extracellular vesicles (EVs) shed by macrophages might have a role in this process. Here, we demonstrated that large EVs released by anti-inflammatory human macrophages decreased PDAC cellular sensitivity to gemcitabine. Using proteomic analysis, chitinase 3-like-1 (CHI3L1) and fibronectin (FN1) were identified as two of the most abundant proteins in the cargo of macrophages-derived EVs. Overexpression of CHI3L1 and FN1, using recombinant human proteins, induced PDAC cellular resistance to gemcitabine through ERK (extracellular-signal-regulated kinase) activation. Inhibition of CHI3L1 and FN1 by pentoxifylline and pirfenidone, respectively, partially reverted gemcitabine resistance. In PDAC patient samples, CHI3L1 and FN1 were expressed in the stroma, associated with the high presence of macrophages. The Cancer Genome Atlas analysis revealed an association between CHI3L1 and FN1 gene expression, overall survival of PDAC patients, gemcitabine response, and macrophage infiltration. Altogether, our data identifies CHI3L1 and FN1 as potential targets for pharmacological inhibition in PDAC. Further pre-clinical in vivo work is warranted to study the possibility of repurposing pentoxifylline and pirfenidone as adjuvant therapies for PDAC treatment.

Sex, Digital Devices, Social Media, and Social Isolation: A Study on Sexual Behavioral During COVID -19 Pandemic.

INTRODUCTION: Coronavirus (COVID-19) pandemic has caused social and economic damages. People have adapted to a new reality of physical distance. OBJECTIVE: The study aimed to assess the use of digital devices and social media, focusing on psychosocial and demographic factors of people´s sexual behavior during the pandemic. METHODS: A total of 1,357 Brazilian adults participated in a cross-sectional online survey. They were recruited through social media to obtain information regarding sexual behavior and the use of digital devices and social media. RESULTS: Digital devices and social media were used by 38.8% of the participants. Among the group that used technological devices, most claimed to have changed their sexual behavior, with 76.9% consuming more sexual content through movies or series. CONCLUSION: In a smaller group, technological resources appeared as an alternative for safer sex, reducing the risks of COVID-19 transmission.

Phenotypic Analysis of Urothelial Exfoliated Cells in Bladder Cancer via Microfluidic Immunoassays: Sialyl-Tn as a Novel Biomarker in Liquid Biopsies.

Bladder cancer is the most common malignancy of the urinary tract, having one of the highest recurrence rates and progression from non-muscle to muscle invasive bladder cancer that commonly leads to metastasis. Cystoscopy and urine cytology are the standard procedures for its detection but have limited clinical sensitivity and specificity. Herein, a microfluidic device, the UriChip, was developed for the enrichment of urothelial exfoliated cells from fresh and frozen urine, based on deformability and size, and the cancer-associated glycan Sialyl-Tn explored as a putative bladder cancer urinary biomarker. Spiking experiments with bladder cancer cell lines showed an isolation efficiency of 53%, while clinical sample analyses revealed retention of cells with various morphologies and sizes. in situ immunoassays demonstrated significantly higher number of Sialyl-Tn-positive cells in fresh and frozen voided urine from bladder cancer patients, compared to healthy individuals. Of note, urothelial exfoliated cells from cryopreserved urine sediments were also successfully isolated by the UriChip, and found to express significantly high levels of Sialyl-Tn. Remarkably, Sialyl-Tn expression is correlated with tumor stage and grade. Overall, our findings demonstrate the potential of UriChip and Sialyl-Tn to detect urothelial bladder cancer cells in follow-up and long-term retrospective studies.

Estrogen receptors in urogenital schistosomiasis and bladder cancer: Estrogen receptor alpha-mediated cell proliferation.

Estrogen-like metabolites have been identified in S. haematobium, the helminth parasite that causes urogenital schistosomiasis (UGS) and in patients´ blood and urine during UGS. Estrogen receptor (ER) activation is enriched in the luminal molecular subtype bladder cancer (BlaCa). To date, the significance of ER to these diseases remains elusive. We evaluated ERα and ERß expression in UGS-related BlaCa (n = 27), UGS-related non-malignant lesions (n = 35), and noninfected BlaCa (n = 80). We investigated the potential of ERα to recognize S. haematobium-derived metabolites by docking and molecular dynamics simulations and studied ERα modulation in vitro using 3 BlaCa cell lines, T24, 5637 and HT1376. ERα was expressed in tumor and stromal cells in approximately 20% noninfected cases and in 30% of UGS-related BlaCa, predominantly in the epithelial cells. Overall, ERα expression was associated with features of tumor aggressiveness such as high proliferation and p53 positive expression. ERα expression correlated with presence of schistosome eggs. ERß was widely expressed in both cohorts but weaker in UGS-related cases. molecular dynamics simulations of the 4 most abundant S. haematobium-derived metabolites revealed that smaller metabolites have comparable affinity for the ERα active state than 17ß-estradiol, while the larger metabolites present higher affinity. Our in vitro findings suggested that ERα activation promotes proliferation in ERα expressing BlaCa cells and that this can be reverted with anti-estrogenic therapy. In summary, we report differential ER expression between UGS-related BlaCa and noninfected BlaCa and provide evidence supporting a role of active ERα during UGS and UGS-induced carcinogenesis.

Preoperative imaging characteristics predict poor survival and inadequate resection for left-sided pancreatic adenocarcinoma: a multi-institutional analysis.

BACKGROUND: Optimal treatment of pancreatic ductal adenocarcinoma of the neck, body and tail (PDAC-NBT) necessitates R0 surgical resection. Preoperative radiographic identification of patients likely to achieve successful oncologic resection remains difficult. This study seeks to identify preoperative imaging characteristics predictive of non-R0 resections or impaired survival for PDAC-NBT. METHODS: Patients at five high-volume centers who underwent resection for PDAC-NBT were retrospectively analyzed. The most immediate preoperative cross-sectional scan was assessed along with outcome measures of overall survival and margin status. RESULTS: 330 patients were treated between 2001 and 2016. Margin status included 247 R0 (78.2%), 67 R1 (21.2%), and 2 R2 (0.6%). A non-R0 resection predicted worse survival (p = 0.0002). On preoperative imaging, patients with tumors greater than 20 mm, tumor attenuation greater than 70 Hounsfield units, or who demonstrated pancreatic atrophy and/or calcifications also had worse survival (p = 0.010, p = 0.036, p = 0.025 respectively). Patients with tumors interfacing with the splenic artery or vein or extending posteriorly achieved fewer R0 resections (p = 0.0006, p = 0.0004, p = 0.001, respectively). CONCLUSION: Preoperative cross-sectional imaging can identify tumor characteristics associated with poor survival and non-R0 resection. Further investigation is needed to identify the appropriate surgical and treatment modifications necessary to clinically benefit this subset of patients.

Competitive glucose metabolism as a target to boost bladder cancer immunotherapy.

Bladder cancer - the tenth most frequent cancer worldwide - has a heterogeneous natural history and clinical behaviour. The predominant histological subtype, urothelial bladder carcinoma, is characterized by high recurrence rates, progression and both primary and acquired resistance to platinum-based therapy, which impose a considerable economic burden on health-care systems and have substantial effects on the quality of life and the overall outcomes of patients with bladder cancer. The incidence of urothelial tumours is increasing owing to population growth and ageing, so novel therapeutic options are vital. Based on work by The Cancer Genome Atlas project, which has identified targetable vulnerabilities in bladder cancer, immune checkpoint inhibitors (ICIs) have arisen as an effective alternative for managing advanced disease. However, although ICIs have shown durable responses in a subset of patients with bladder cancer, the overall response rate is only ~15-25%, which increases the demand for biomarkers of response and therapeutic strategies that can overcome resistance to ICIs. In ICI non-responders, cancer cells use effective mechanisms to evade immune cell antitumour activity; the overlapping Warburg effect machinery of cancer and immune cells is a putative determinant of the immunosuppressive phenotype in bladder cancer. This energetic interplay between tumour and immune cells leads to metabolic competition in the tumour ecosystem, limiting nutrient availability and leading to microenvironmental acidosis, which hinders immune cell function. Thus, molecular hallmarks of cancer cell metabolism are potential therapeutic targets, not only to eliminate malignant cells but also to boost the efficacy of immunotherapy. In this sense, integrating the targeting of tumour metabolism into immunotherapy design seems a rational approach to improve the therapeutic efficacy of ICIs.

Survival from cancer in the north region of Portugal: results from the first decade of the millennium.

The aim of this study was to evaluate net survival from cancer diagnosed during the period 2001-2010 in the north region of Portugal to identify the tumours that need actions to improve the outcomes. Data were retrieved from the North Region Cancer Registry of Portugal database. The top 20 cancer sites in adults were considered: oesophagus, stomach, colon, rectum, pancreas, liver, larynx, lung, skin melanoma, breast, cervix, corpus uteri, ovary, prostate, kidney, bladder, brain and central nervous system, thyroid, non-Hodgkin lymphoma and multiple myeloma. Net survival was estimated using the Pohar-Perme estimator. The effect of diagnosis period was evaluated using flexible parametric models adjusted for age and sex where appropriate. Thyroid and prostate cancers presented the best 5-year survival (>90%), whereas oesophagus, pancreas, liver and lung cancers the worst 5-year survival (<20%). The largest increase in survival was observed for the larynx. A significant decrease in age-adjusted and sex-adjusted excess mortality was observed for stomach, colon, pancreas, larynx, melanoma, breast, brain and central nervous system, thyroid, non-Hodgkin lymphoma and multiple myeloma. For the other cancer sites, no significant trends were observed. For some of these sites, the downward trend in excess mortality was only observed in the short term. An important picture of population-based cancer survival outcomes for the first decade of the millennium in the north region of Portugal was presented in this study. It has been shown that improvements in survival were not universal for all cancer sites. These results should be used to highlight tumours where intervention is needed the most.

Incidencia de fracturas patológicas por metástasis en el fémur proximal en un hospital de concentración / Incidence of proximal femur pathologic fractures secondary to metastasis in a national reference orthopedic hospital

Resumen: Objetivo: Reportar la incidencia de fracturas en terreno patológico secundarias a metástasis que se presentaron en un hospital de referencia nacional en un período de cinco años. Métodos: Se registraron los ingresos totales a nuestro centro, así como los pacientes que satisficieran la condición de presentar una fractura de fémur proximal en terreno patológico. Con base en información oficial, se calculó el tamaño de población derechohabiente potencial de acuerdo al área de influencia. Con base en los datos se hizo el cálculo de la incidencia anual y de la densidad de incidencia. Resultados: Se identificaron 98 fracturas en 95 individuos. El cálculo de la densidad de incidencia fue de 0.70/100,000 personas durante el período de observación. Se incluyeron 54 sujetos femeninos y 41 masculinos con un promedio de edad de 65.3 años, aunque el rango fue muy variable (de 18 a 90 años). La mayor parte de las personas presentó metástasis por tumores sólidos. De los casos, 29% fue tratado de manera conservadora y el resto requirió tratamiento quirúrgico que incluyó desde osteosíntesis hasta artroplastía protésica. El promedio de estancia hospitalaria fue ligeramente mayor a una semana. Conclusiones: La incidencia reportada es relativamente baja. Encontramos una gran variedad de orígenes y localizaciones anatómicas. No se puede, al momento, generalizar tratamientos o predecir supervivencia.

Abstract: Objective: To report the incidence of pathological fractures secondary to metastasis at a national referral hospital during a 5-year period. Methods: Total admissions to our center were recorded, together with the patients who met the requirement of having a proximal femur fracture in a pathological area. The potential number of beneficiaries was estimated based on official figures and the hospital's area of influence. The annual incidence rate and the incidence density were calculated using the latter data. Results: 98 fractures were identified in 95 patients. The calculated incidence density was 0.70/100,000 population during the observation period. Fifty-four female patients and 41 male patients were included. Mean age was 65.3 years, with a very wide age range (18-90 years). Most patients had metastasis of solid tumors. Twenty-nine percent of patients were treated conservatively and the rest of them required surgery that included from osteosynthesis to prosthetic arthroplasty. The mean length of stay was over one week. Conclusions: The reported incidence of this type of fractures is relatively low. We found a wide variety of anatomical origins and locations. As of now, it is not possible to generalize the treatment or predict the survival.

Glycomic analysis of gastric carcinoma cells discloses glycans as modulators of RON receptor tyrosine kinase activation in cancer.

BACKGROUND: Terminal α2-3 and α2-6 sialylation of glycans precludes further chain elongation, leading to the biosynthesis of cancer relevant epitopes such as sialyl-Lewis X (SLe(X)). SLe(X) overexpression is associated with tumor aggressive phenotype and patients' poor prognosis. METHODS: MKN45 gastric carcinoma cells transfected with the sialyltransferase ST3GAL4 were established as a model overexpressing sialylated terminal glycans. We have evaluated at the structural level the glycome and the sialoproteome of this gastric cancer cell line applying liquid chromatography and mass spectrometry. We further validated an identified target expression by proximity ligation assay in gastric tumors. RESULTS: Our results showed that ST3GAL4 overexpression leads to several glycosylation alterations, including reduced O-glycan extension and decreased bisected and increased branched N-glycans. A shift from α2-6 towards α2-3 linked sialylated N-glycans was also observed. Sialoproteomic analysis further identified 47 proteins with significantly increased sialylated N-glycans. These included integrins, insulin receptor, carcinoembryonic antigens and RON receptor tyrosine kinase, which are proteins known to be key players in malignancy. Further analysis of RON confirmed its modification with SLe(X) and the concomitant activation. SLe(X) and RON co-expression was validated in gastric tumors. CONCLUSION: The overexpression of ST3GAL4 interferes with the overall glycophenotype of cancer cells affecting a multitude of key proteins involved in malignancy. Aberrant glycosylation of the RON receptor was shown as an alternative mechanism of oncogenic activation. GENERAL SIGNIFICANCE: This study provides novel targets and points to an integrative tumor glycomic/proteomic-profiling for gastric cancer patients' stratification. This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc.

Metabolic coupling in urothelial bladder cancer compartments and its correlation to tumor aggressiveness.

Monocarboxylate transporters (MCTs) are vital for intracellular pH homeostasis by extruding lactate from highly glycolytic cells. These molecules are key players of the metabolic reprogramming of cancer cells, and evidence indicates a potential contribution in urothelial bladder cancer (UBC) aggressiveness and chemoresistance. However, the specific role of MCTs in the metabolic compartmentalization within bladder tumors, namely their preponderance on the tumor stroma, remains to be elucidated. Thus, we evaluated the immunoexpression of MCTs in the different compartments of UBC tissue samples (n = 111), assessing the correlations among them and with the clinical and prognostic parameters. A significant decrease in positivity for MCT1 and MCT4 occurred from normoxic toward hypoxic regions. Significant associations were found between the expression of MCT4 in hypoxic tumor cells and in the tumor stroma. MCT1 staining in normoxic tumor areas, and MCT4 staining in hypoxic regions, in the tumor stroma and in the blood vessels were significantly associated with UBC aggressiveness. MCT4 concomitant positivity in hypoxic tumor cells and in the tumor stroma, as well as positivity in each of these regions concomitant with MCT1 positivity in normoxic tumor cells, was significantly associated with an unfavourable clinicopathological profile, and predicted lower overall survival rates among patients receiving platinum-based chemotherapy. Our results point to the existence of a multi-compartment metabolic model in UBC, providing evidence of a metabolic coupling between catabolic stromal and cancer cells' compartments, and the anabolic cancer cells. It is urgent to further explore the involvement of this metabolic coupling in UBC progression and chemoresistance.

[Pathological fracture rate due to proximal femur metastasis at a referral hospital]. / Incidencia de fracturas patológicas por metástasis en el fémur proximal en un hospital de concentración.

OBJECTIVE: To report the incidence of pathological fractures secondary to metastasis at a national referral hospital during a 5-year period. METHODS: Total admissions to our center were recorded, together with the patients who met the requirement of having a proximal femur fracture in a pathological area. The potential number of beneficiaries was estimated based on official figures and the hospitals area of influence. The annual incidence rate and the incidence density were calculated using the latter data. RESULTS: 98 fractures were identified in 95 patients. The calculated incidence density was 0.70/100,000 population during the observation period. Fifty-four female patients and 41 male patients were included. Mean age was 65.3 years, with a very wide age range (18-90 years). Most patients had metastasis of solid tumors. Twenty-nine percent of patients were treated conservatively and the rest of them required surgery that included from osteosynthesis to prosthetic arthroplasty. The mean length of stay was over one week. CONCLUSIONS: The reported incidence of this type of fractures is relatively low. We found a wide variety of anatomical origins and locations. As of now, it is not possible to generalize the treatment or predict the survival.

Reportar la incidencia de fracturas en terreno patológico secundarias a metástasis que se presentaron en un hospital de referencia nacional en un período de cinco años.

Reducción cerrada de la articulación sacroilíaca y reconstrucción acetabular en un paciente pediátrico con fractura inestable de pelvis Torode & Zieg IVd. Reporte de un caso / Closed reduction of the sacroiliac joint and acetabular reconstruction in a pediatric patient with a Torode/Zieg IVd unstable pelvic fracture. Case report

Resumen: Antecedentes: Las lesiones pélvicas severas por alta energía en pacientes pediátricos son raras, y más, asociadas con fracturas acetabulares, siendo lesiones que ponen en riesgo la vida del niño. Método: Se presenta el caso de un individuo pediátrico con diagnóstico de fractura de pelvis Torode y Zieg tipo IVd con datos clínicos y radiográficos revisados retrospectivamente. Caso clínico: Niño de 12 años de edad politraumatizado, con fractura de pelvis inestable, tratado de manera multidisciplinaria. Se realizó estabilización en la unidad de terapia intensiva, laparotomía por el Servicio de Cirugía General, manejo ortopédico con reducción cerrada, fijación interna de la fractura-luxación sacroilíaca, con posterior reducción abierta y fijación interna de la fractura acetabular derecha; presentó adecuada evolución clínica. Conclusiones: El diagnóstico de una fractura de acetábulo Torode y Zieg IVd es una indicación absoluta de manejo quirúrgico. La atención multidisciplinaria es esencial para preservar la vida de los sujetos con lesiones de pelvis inestable. Este caso muestra la complejidad de una fractura pélvica inestable asociada con múltiples lesiones debido a un mecanismo poco frecuente (aplastamiento por un árbol) en un paciente con un sistema esquelético inmaduro.

Abstract: Background: High-energy severe pelvic injuries in pediatric patients are rare, particularly those associated with acetabular fractures. They are life threatening injuries. Method: We report the case of a pediatric patient with a diagnosis of a Torode/Zieg IVd pelvic fracture. The clinical and radiographic data was reviewed retrospectively. Clinical case: Male, 12 year-old polytraumatized patient with an unstable pelvic fracture who underwent multidisciplinary treatment. He was stabilized in the intensive care unit; the general surgery service performed laparotomy; orthopedic management consisted of closed reduction and internal fixation of the sacroiliac fracture-dislocation and later of open reduction and internal fixation of the right acetabular fracture. The patient´s clinical course was appropriate. Conclusions: A diagnosis of a type IVd Torode/Zieg fracture is an absolute indication for surgical management. A multidisciplinary approach is essential to save the life of patients with unstable pelvic lesions. This case shows the complexity of an unstable pelvic fracture associated with multiple injuries, due to a rather infrequent mechanism of injury (being crushed by a tree) in a patient with an immature skeletal system.