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Abstract Deep single-cell multi-omic profiling offers a promising approach to understand and overcome drug resistance in relapsed or refractory (rr) acute myeloid leukemia (AML). Here, we combine single-cell ex vivo drug profiling (pharmacoscopy) with single-cell and bulk DNA, RNA, and protein analyses, alongside clinical data from 21 rrAML patients. Unsupervised data integration reveals reduced ex vivo response to the Bcl-2 inhibitor venetoclax (VEN) in patients treated with both a hypomethylating agent (HMA) and VEN, compared to those pre-exposed to chemotherapy or HMA alone. Integrative analysis identifies both known and unreported mechanisms of innate and treatment-related VEN resistance and suggests alternative treatments, like targeting increased proliferation with the PLK inhibitor volasertib. Additionally, high CD36 expression in VEN-resistant blasts associates with sensitivity to CD36-targeted antibody treatment ex vivo. This study demonstrates how single-cell multi-omic profiling can uncover drug resistance mechanisms and treatment vulnerabilities, providing a valuable resource for future AML research.

Summary: Stemness and pluripotency are mediated by transcriptional master regulators that promote self-renewal and repress cell differentiation, among which is the high-mobility group (HMG) box transcription factor SOX2. Dysregulated SOX2 expression, by contrast, leads to transcriptional aberrations relevant to oncogenic transformation, cancer progression, metastasis, therapy resistance, and relapse. Here, we report a post-transcriptional mechanism by which the cytosolic pool of SOX2 contributes to these events in an unsuspected manner. Specifically, a low-complexity region within SOX2’s C-terminal segment connects to the ribosome to modulate the expression of cognate downstream factors. Independent of nuclear structures or DNA, this C-terminal functionality alone changes metabolic properties and induces non-adhesive growth when expressed in the cytosol of SOX2 knockout cells. We thus propose a revised model of SOX2 action where nuclear and cytosolic fractions cooperate to impose cell fate decisions via both transcriptional and translational mechanisms.

Summary: Three-dimensional (3D) ex vivo cultures allow the study of cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring on-target drug sensitivity in a scaffold-free 3D culture system through quantification of apoptotic tumor cells. We provide detailed steps for sample processing, immunofluorescence staining, semi-high-throughput confocal imaging, and imaged-based quantification of 3D cultures. This protocol is versatile and can be applied in principle to any patient-derived material. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract Intratumoral tertiary lymphoid structures (TLSs) have been associated with improved outcome in various cohorts of patients with cancer, reflecting their contribution to the development of tumor-targeting immunity. Here, we demonstrate that high-grade serous ovarian carcinoma (HGSOC) contains distinct immune aggregates with varying degrees of organization and maturation. Specifically, mature TLSs (mTLS) as forming only in 16% of HGSOCs with relatively elevated tumor mutational burden (TMB) are associated with an increased intratumoral density of CD8+ effector T (TEFF) cells and TIM3+PD1+, hence poorly immune checkpoint inhibitor (ICI)-sensitive, CD8+ T cells. Conversely, CD8+ T cells from immunologically hot tumors like non-small cell lung carcinoma (NSCLC) are enriched in ICI-responsive TCF1+ PD1+ T cells. Spatial B-cell profiling identifies patterns of in situ maturation and differentiation associated with mTLSs. Moreover, B-cell depletion promotes signs of a dysfunctional CD8+ T cell compartment among tumor-infiltrating lymphocytes from freshly isolated HGSOC and NSCLC biopsies. Taken together, our data demonstrate that – at odds with NSCLC – HGSOC is associated with a low density of follicular helper T cells and thus develops a limited number of mTLS that might be insufficient to preserve a ICI-sensitive TCF1+PD1+ CD8+ T cell phenotype. These findings point to key quantitative and qualitative differences between mTLSs in ICI-responsive vs ICI-irresponsive neoplasms that may guide the development of alternative immunotherapies for patients with HGSOC.

IntroductionThe pathogenic bacterium Helicobacter pylori has evolved glycan-mediated mechanisms to evade host immune defenses. This study tests the hypothesis that genetic disruption of H. pylori glycan biosynthesis alters immune recognition and response by human gastric epithelial cells and monocyte-derived dendritic cells.MethodsTo test this hypothesis, human cell lines were challenged with wildtype H. pylori alongside an array of H. pylori glycosylation mutants. The relative levels of immune response were measured via immature dendritic cell maturation and cytokine secretion.ResultsOur findings indicate that disruption of lipopolysaccharide biosynthesis diminishes gastric cytokine production, without disrupting dendritic cell recognition and activation. In contrast, variable immune responses were observed in protein glycosylation mutants which prompted us to test the hypothesis that phase variation plays a role in regulating bacterial cell surface glycosylation and subsequent immune recognition. Lewis antigen presentation does not correlate with extent of immune response, while the extent of lipopolysaccharide O-antigen elaboration does.DiscussionThe outcomes of this study demonstrate that H. pylori glycans modulate the host immune response. This work provides a foundation to pursue immune-based tailoring of bacterial glycans towards modulating immunogenicity of microbial pathogens.

Abstract Understanding the complex background of cancer requires genotype-phenotype information in single-cell resolution. Here, we perform long-read single-cell RNA sequencing (scRNA-seq) on clinical samples from three ovarian cancer patients presenting with omental metastasis and increase the PacBio sequencing depth to 12,000 reads per cell. Our approach captures 152,000 isoforms, of which over 52,000 were not previously reported. Isoform-level analysis accounting for non-coding isoforms reveals 20% overestimation of protein-coding gene expression on average. We also detect cell type-specific isoform and poly-adenylation site usage in tumor and mesothelial cells, and find that mesothelial cells transition into cancer-associated fibroblasts in the metastasis, partly through the TGF-β/miR-29/Collagen axis. Furthermore, we identify gene fusions, including an experimentally validated IGF2BP2::TESPA1 fusion, which is misclassified as high TESPA1 expression in matched short-read data, and call mutations confirmed by targeted NGS cancer gene panel results. With these findings, we envision long-read scRNA-seq to become increasingly relevant in oncology and personalized medicine.

Comme il convient à une conférence d’ouverture. cette étude sera avant tout introductive. Je ne renoncerai pourtant pas à un pouvoir de proposition sur deux questions

Abstract Cancer patients with advanced disease are characterized by intrinsic challenges in predicting drug response patterns, often leading to ineffective treatment. Current clinical practice for treatment decision-making is commonly based on primary or secondary tumour biopsies, yet when disease progression accelerates, tissue biopsies are not performed on a regular basis. It is in this context that liquid biopsies may offer a unique window to uncover key vulnerabilities, providing valuable information about previously underappreciated treatment opportunities. Here, we present MyCTC chip, a novel microfluidic device enabling the isolation, culture and drug susceptibility testing of cancer cells derived from liquid biopsies. Cancer cell capture is achieved through a label-free, antigen-agnostic enrichment method, and it is followed by cultivation in dedicated conditions, allowing on-chip expansion of captured cells. Upon growth, cancer cells are then transferred to drug screen chambers located within the same device, where multiple compounds can be tested simultaneously. We demonstrate MyCTC chip performance by means of spike-in experiments with patient-derived breast circulating tumour cells, enabling >95% capture rates, as well as prospective processing of blood from breast cancer patients and ascites fluid from patients with ovarian, tubal and endometrial cancer, where sensitivity to specific chemotherapeutic agents was identified. Together, we provide evidence that MyCTC chip may be used to identify personalized drug response patterns in patients with advanced metastatic disease and with limited treatment opportunities.

Abstract PARP inhibitors (PARPi) have revolutionized the therapeutic landscape of epithelial ovarian cancer (EOC) treatment with outstanding benefits in regard to progression-free survival, especially in patients either carrying BRCA1/2 mutations or harboring defects in the homologous recombination repair system. Yet, it remains uncertain which PARPi to apply and how to predict responders when platinum sensitivity is unknown. To shed light on the predictive power of genes previously suggested to be associated with PARPi response, we systematically reviewed the literature and identified 79 publications investigating a total of 93 genes. The top candidate genes were further tested using a comprehensive CRISPR-Cas9 mutagenesis screening in combination with olaparib treatment. Therefore, we generated six constitutive Cas9 + EOC cell lines and profiled 33 genes in a CRISPR-Cas9 cell competition assay using non-essential (AAVS1) and essential (RPA3 and PCNA) genes for cell fitness as negative and positive controls, respectively. We identified only ATM, MUS81, NBN, BRCA2, and RAD51B as predictive markers for olaparib response. As the major survival benefit of PARPi treatment was reported in platinum-sensitive tumors, we next assessed nine top candidate genes in combination with three PARPi and carboplatin. Interestingly, we observed similar dropout rates in a gene and compound independent manner, supporting the strong correlation of cancer cell response to compounds that rely on DNA repair for their effectiveness. In addition, we report on CDK12 as a common vulnerability for EOC cell survival and proliferation without altering the olaparib response, highlighting its potential as a therapeutic target in EOC.

Objectives: Liberia is endemic to Lassa fever (LF) and has the largest reported per capita incidence of LF patients in the West African region. Cases of the disease increased unprecedentedly in 2019 and 2020, characterized by a geographical drift in epidemiology and seasonal variation of occurrence. This study aims to describe the epidemiological and clinical characteristics of LF in Liberia from 2019 to 2020. Methods: A retrospective study was conducted on cases of LF confirmed at the National Public Health Reference Laboratory from January 2019 to December 2020. Medical records were reviewed, and epidemiological and clinical data were collected in an organized manner. Descriptive and inferential statistics were carried out using Epi Info (version 7.2.5.0). Results: A total of 382 suspected LF cases were reported, of which 103 were laboratory-confirmed, yielding a case positivity rate of 27% (103/382). The median age of the LF cases was 20 (IQR: 9–30). Children younger than 18 years accounted for 40.8% (42/103) of the cases and healthcare workers’ cases constituted 7.7% of the cases. Bong, Nimba, and Grand Bass accounted for 87.4% of the cases with cases in new counties like Lofa, Margibi, and Grand Kru. Hemorrhage (aOR:10.2; 95% CI: 3.11–33.81), patients who did not receive ribavirin (aOR: 4.4; 95% CI: 1.12–17.57, P = 0.034), and patients aged 40 years or older (aOR: 6.2; 95% CI: 1.19–32.53, P = 0.049) were associated with LF mortality. Conclusion: The LF cases in 2019 and 2020 had a high case fatality rate and spread to new counties that had not previously reported LF. The disease occurred during most of the rainy season instead of the usual dry season. There is an urgent need to lower morbidity and mortality, improve early presentation to the hospital, and early initiation of appropriate medical care.

Summary: We provide a protocol using non-homologous end joining to integrate an oligonucleotide sequence of a fluorescence protein at the CDH1 locus encoding for the epithelial glycoprotein E-cadherin. We describe steps for implementing the CRISPR-Cas9-mediated knock-in procedure by transfecting a cancer cell line with a pool of plasmids. The EGFP-tagged cells are traced by fluorescence-activated cell sorting and validated on DNA and protein levels. The protocol is flexible and can be applied in principle to any protein expressed in a cell line.For complete details on the use and execution of this protocol, please refer to Cumin et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

During the COVID 19 pandemic, round-the-clock demand for COVID -19 laboratory tests exceeded capacity, placing a significant burden on laboratory staff and infrastructure. The use of laboratory information management systems (LIMS) to streamline all phases of laboratory testing (preanalytical, analytical, and postanalytical) has become inevitable. The objective of this study is to describe the architecture, implementation, and requirements of PlaCARD, a software platform for managing patient registration, medical specimens, and diagnostic data flow, as well as reporting and authentication of diagnostic results during the 2019 coronavirus pandemic (COVID -19) in Cameroon. Building on its experience with biosurveillance, CPC developed an open-source, real-time digital health platform with web and mobile applications called PlaCARD to improve the efficiency and timing of disease-related interventions. PlaCARD was quickly adapted to the decentralization strategy of the COVID 19 testing in Cameroon and, after specific user training, was deployed in all COVID 19 diagnostic laboratories and the regional emergency operations center. Overall, 71% of samples tested for COVID 19 by molecular diagnostics in Cameroon from 05 March 2020 to 31 October 2021 were entered into PlaCARD. The median turnaround time for providing results was 2 days [0-2.3] before April 2021 and decreased to 1 day [1- 1] after the introduction of SMS result notification in PlaCARD. The integration of LIMS and workflow management into a single comprehensive software platform (PlaCARD) has strengthened COVID 19 surveillance capabilities in Cameroon. PlaCARD has demonstrated that it can be used as a LIMS for managing and securing test data during an outbreak.

Geophysical survey for groundwater was conducted in two newly built halls of residence (hostels) named Dangote and Shehu Idris at the second phase of Ahmadu Bello University (ABU), Zaria in Nigeria. The vertical electrical sounding (VES) technique of resistivity method was employed with Schlumberger array in seven VES points distributed within the two hostels. Results showed that 4 to 6 geoelectric layers exist within the study area for a spacing of at most 200 m between current electrodes. The resistivity values of the geoelectric layers in all the VES points ranged from 26.6 Ωm – 30342 Ωm while the thickness and depth ranged from 0.263 m – 40.34 m and 0.572 m – 50.66 m respectively. However, four VES points (two in each hostel) were identified to be suitable for borehole drilling though, three out of these four VES points have single aquifer in each with resistivity and thickness ranging from 34.4 Ωm – 40.57 Ωm and 2.02 m – 7.49 m respectively. Conversely, the fourth VES point has two aquifers positioned at the third and fifth layers with resistivity values ranging from 26.6 Ωm – 51.4 Ωm and thickness ranging from 1.93 m – 18.7 m. It was recommended that among the four VES points (two in each hostel) identified to contain aquifers, priority should be given to the ones with larger thickness in each hostel.

The unhairing stage of leather processing is associated with the production of significant amounts of solid and liquid wastes. The use of enzymes to replace the polluting sulphides currently used for unhairing is a viable alternative. Various proteases from different Bacillus cereus strains as well as many other bacterial strains have been used successfully for the unhairing of skins. However, no previous work has assessed the use of the crude alkaline protease extract from Bacillus cereus strain 1-p, a novel Bacillus cereus strain obtained from the shores of Lake Bogoria - a soda lake in Kenya – in the unhairing of goatskins. This study, therefore, evaluates the potential of the protease extract from the Bacillus cereus strain 1-p to unhair goatskins. Optimum variables for unhairing using the protease were investigated. Complete unhairing was achieved within 12 hours at 27°C and pH 12 using the crude enzyme. The period and temperature required for complete unhairing were significantly lower than that of other enzymatic unhairing techniques. Compared to the leather unhaired with sulphide, the leather unhaired with the enzyme did not only show superior organoleptic properties but also recorded comparable or superior physical properties, namely tensile strength (26.94 N/mm2), percentage elongation (76.29%), tear strength (43.59 N/mm), and distension at grain crack and burst (7.9 mm and 8.2 mm respectively). The wastewater from the enzymatic unhairing process recorded a significant reduction in biochemical oxygen demand (78%), chemical oxygen demand (83%), and the wastewater volume (50%) compared to the process that uses sulphide. It was concluded that the use of the crude protease extract from the Bacillus cereus strain 1-p in unhairing goatskins is feasible.

In the era of personalized medicine, where transition from organ‐based to individualized genetic diagnosis takes place, the tailoring of treatment in cancer becomes increasingly important. This is particularly true for high‐grade, advanced FIGO stage serous adenocarcinomas of the ovary (OC), fallopian tube (TC), and peritoneum (PC), which are currently all treated identically. We analyzed three independent patient cohorts using histopathologically classified diagnosis and various molecular approaches (transcriptomics, immunohistochemistry, next‐generation sequencing, fluorescent and chromogenic in situ hybridization). Using multivariate Cox regression model, we found that PC is more aggressive compared with advanced‐stage OC independent of residual disease as shown by an earlier relapse‐free survival in two large cohorts (HR: 2.63, CI: 1.59–4.37, P

Superoxide dismutase 2 (SOD2) is an antioxidant enzyme that appears phylogenetically conserved. However, functional Sod2 polymorphisms have been studied, and the specific polymorphisms are related to activity alterations of the SOD2 enzyme. An example of a polymorphism of SOD2 is Val16Ala (rs4880), which has been identified in exon 2 of the human Sod2 gene. This polymorphism is recognized as a single nucleotide polymorphism (SNP) and alters the conformation of SOD2. Additionally, recent studies have shown that the Ala16 Val polymorphism in Sod2 can be related to different pathological diseases. In these terms, the objective of the present study was to evaluate whether the polymorphism of SOD2 in Val16Ala (rs4880) influences the motility and vigor of X- and Y-bearing sperm at different pH values promoting sperm selection. We found that polymorphism rs4880 at normal pH conditions can result in alterations in the activity of superoxide dismutase in the sperm through different assay analyses. Moreover, compelling modulation evidence indicates that this effect could also mediate seminal plasma redox alterations and consequently can play an important role in sperm physiology, fertilization, and postfertilization.

The bodies of both Kha and Merit were wrapped in a wide linen sheet stitched along the bottom; but Merit had a second funerary sheet and a painted mask. The condition of the textiles was very poor due to both age and long display. There was a general lack of strength and elasticity in the fibers which were fragile, very brittle and partly covered by dust. In order to consolidate the textile of Merit, it was necessary to temporarily remove the mask and second sheet, which were conserved separately. After vacuum cleaning the mummies, consolidation was achieved by sewing a dyed nylon net around the bodies.

Summary: Integrin alpha 2 (ITGA2) promotes cancer metastasis through selective adhesion to ECM proteins; however, the specific contribution of integrin glycosylation remains uncertain. We provide evidence that ITGA2 is a highly glycosylated transmembrane protein expressed in ovarian cancer tissue and cell lines. In-depth glycoproteomics identified predominant N- and O-glycosylation sites harboring substantially divergent ITGA2 glycosylation profiles. Generated putative ITGA2 N-glycosite mutants halted collagen and laminin binding and cells lacking N-glycosylated ITGA2 were marginally adherent to collagen, likely associated with its enhanced proteasome degradation through poly-ubiquitination. Proteomic and enrichment pathway analysis revealed increased cellular apoptosis and collagen organization in non-glycosylated ITGA2 mutant cells. Moreover, we provide evidence that ITGA2-specific sialylation is involved in selective cell-ECM binding. These results highlight the importance of glycans in regulating ITGA2 stability and ligand binding capacity which in turn modulates downstream focal adhesion and promotes cell survival in a collagen environment.

Introdução: O atendimento em Saúde da Criança por médicos da Estratégia de Saúde da Família (ESF) fundamenta-se em recomendações do Ministério da Saúde e da Sociedade Brasileira de Medicina de Família e Comunidade. Objetivo: Identificar as dificuldades dos médicos da ESF para ofertarem assistência em Saúde da Criança no município de Patos de Minas/MG, já que a assistência na Atenção Básica (AB) é prestada pela ESF em quase todo o território neste município. Métodos: Foi aplicado questionário estruturado aos 42 médicos que atuam na ESF, com questões específicas relacionadas ao perfil profissional e da equipe da ESF e quanto às ações em Saúde da Criança na anamnese, exame físico, exames laboratoriais e orientações. Resultados: Responderam ao questionário 32 médicos (76%). O principal motivo para estarem na ESF foi o interesse pelo trabalho na AB (78%). As dificuldades mais frequentes relacionadas à equipe foram a falta de realização dos grupos de puericultura (41%) e a ausência da puericultura compartilhada com o enfermeiro (41%). Apontaram que as habilidades clínicas e a abordagem familiar são fatores que dificultam a realização da consulta de puericultura. As dificuldades apresentadas pelos médicos foram quanto às habilidades no exame físico do recém-nascido, principalmente exame dos olhos (59%), audição (63%) e avaliação neurológica (53%). Dentre os testes oftalmológicos a dificuldade variou de 69% para acuidade visual até 94% para o teste de cobertura. Dificuldades relacionadas à estrutura física inadequada e/ou inexistência de equipamento ocorreram para a realização do Teste do Olhinho (34%) e para a aferição de pressão arterial (69%). Dificuldades devidas ao desconhecimento de recomendações do Ministério da Saúde ocorreram com o rastreamento de anemia (16%) e do perfil lipídico (41%). Quanto às orientações 16% dos médicos não abordam as curvas de Índice de Massa Corporal (IMC) com as famílias, assim como 19% não abordam violência na infância. As dificuldades identificadas nos médicos quanto às habilidades clínicas, podem estar relacionadas à qualidade de formação na graduação, assim como à ausência ou ineficácia de educação permanente durante o exercício profissional. Conclusão: A assistência à criança em Patos de Minas apresenta falhas, que estão relacionadas a fragilidades dos médicos, à estrutura física e de equipamentos e às equipes da ESF.

Brachytherapy (BT) for locally advanced cervical cancer is vital for optimal outcomes. There is heterogeneity in brachytherapy treatment practice for cervical cancer across India. In an attempt to standardize various processes involved in cervical cancer brachytherapy, the expert members of the Indian Brachytherapy Society (IBS) developed a document related to radiation therapy treatment of cervical cancer with special emphasis on brachytherapy. The guidelines are based on high quality clinical evidence, expert opinion and consensus wherever evidence was lacking. The document provides a guide for external beam radiation and details of all the processes involved in high-dose-rate (HDR) brachytherapy including patient selection, preparation, principles and technique of BT applications, target and normal tissue definition, dose prescriptions, BT planning, reporting parameters, common complications of BT and their management, scope for research, etc. In summary, we present here practical tips and tricks, recording and reporting of cervical cancer brachytherapy, which can be implemented in various clinical environments and forms the basis of this report.

The extracellular matrix (ECM) plays critical roles in tumor progression and metastasis. However, the contribution of ECM proteins to early metastatic onset in the peritoneal cavity remains unexplored. Here, we suggest a new route of metastasis through the interaction of integrin alpha 2 (ITGA2) with collagens enriched in the tumor coinciding with poor outcome in patients with ovarian cancer. Using multiple gene-edited cell lines and patient-derived samples, we demonstrate that ITGA2 triggers cancer cell adhesion to collagen, promotes cell migration, anoikis resistance, mesothelial clearance, and peritoneal metastasis in vitro and in vivo. Mechanistically, phosphoproteomics identify an ITGA2-dependent phosphorylation of focal adhesion kinase and mitogen-activated protein kinase pathway leading to enhanced oncogenic properties. Consequently, specific inhibition of ITGA2-mediated cancer cell-collagen interaction or targeting focal adhesion signaling may present an opportunity for therapeutic intervention of metastatic spread in ovarian cancer.

Ovarian cancer (OC) is one of the most lethal gynecologic malignancies. Due to the lack of specific symptoms and screening methods, this disease is usually diagnosed only at an advanced and metastatic stage. The gold-standard treatment for OC patients consists of debulking surgery followed by taxane combined with platinum-based chemotherapy. Most patients show complete clinical remission after first-line therapy, but the majority of them ultimately relapse, developing radio- and chemoresistant tumors. It is now proposed that the cause of recurrence and reduced therapy efficacy is the presence of small populations of cancer stem cells (CSCs). These cells are usually resistant against conventional cancer therapies and for this reason, effective targeted therapies for the complete eradication of CSCs are urgently needed. In this review article, we highlight the mechanisms of CSC therapy resistance, epithelial-to-mesenchymal transition, stemness, and novel therapeutic strategies for ovarian CSCs.

The presence of ascites in the peritoneal cavity leads to morphological and functional changes of the peritoneal mesothelial cell layer. Cells loose cell-cell interactions, rearrange their cytoskeleton, activate the production of fibronectin, and change their cell surface morphology in a proinflammatory environment. Moreover, ovarian cancer cell adhesion has been shown to be facilitated by these changes due to increased integrin- and CD44-mediated binding sites. In this study, the biological responsiveness of the human pleural mesothelial cell line MeT-5A to patient-derived and artificial ascites was studied in vitro and adhesion of ovarian cancer cells, i.e. SKOV-3 cells, investigated. Changes were mainly observed in cells exposed to artificial ascites containing higher cytokine concentrations than patient-derived ascites. Interestingly, reduced cell-cell interactions were already observed in untreated MeT-5A cells and effects on tight junction protein expression and permeability upon exposure to ascites were minor. Ascites induced upregulation of CDC42 effector protein 2 expression, which affects stress fiber formation, however significant F-actin reorganization was not observed. Moreover, fibronectin production remained unchanged. Analysis of mesothelial cell surface characteristics showed upregulated expression of intercellular adhesion molecule 1, slightly increased hyaluronic acid secretion and decreased microvillus expression upon exposure to ascites. Nevertheless, the observed changes were not sufficient to facilitate adhesion of SKOV-3 cells on MeT-5A cell layer. This study revealed that MeT-5A cells show a reduced biological responsiveness to the presence of ascites, in contrast to published studies on primary human peritoneal mesothelial cells.

El presente artículo analiza cómo la dinámica interventora del Estado colombiano en las actividades económicas, ha incidido en el incremento de consultas populares en temas mineros. Ésta se presenta como un instrumento mediante el cual la comunidad hace frente al otorgamiento desproporcionado de títulos mineros que no cuentan con el requisito de participación de la comunidad afectada o benefciada. Además, se proponen estrategias que el Estado debería adoptar para garantizar el bien común y el interés general acorde con los fnes constitucionales de guiar la economía bajo parámetros del desarrollo sostenible.

Abstract Background The relationship between various external agents such as pollen, food, and infectious agents and human sensitivity exists and is variable depending upon individual’s health conditions. For example, we believe that the pathogenetic potential of the Merkel cell polyomavirus (MCPyV), the resident virus in skin, is variable and depends from the degree of individual’s reactivity. MCPyV as well as Epstein-Barr virus, which are normally connected with humans under the form of subclinical infection, are thought to be involved at various degrees in several neoplastic and inflammatory diseases. In this review, we cover two types of Langerhans cell neoplasms, the Langerhans cell sarcoma (LCS) and Langerhans cell histiocytosis (LCH), represented as either neoplastic or inflammatory diseases caused by MCPyV. Methods We meta-analyzed both our previous analyses, composed of quantitative PCR for MCPyV-DNA, proteomics, immunohistochemistry which construct IL-17 endocrine model and interleukin-1 (IL-1) activation loop model, and other groups’ data. Results We have shown that there were subgroups associated with the MCPyV as a causal agent in these two different neoplasms. Comparatively, LCS, distinct from the LCH, is a neoplastic lesion (or sarcoma) without presence of inflammatory granuloma frequently observed in the elderly. LCH is a proliferative disease of Langerhans-like abnormal cells which carry mutations of genes involved in the RAS/MAPK signaling pathway. We found that MCPyV may be involved in the development of LCH. Conclusion We hypothesized that a subgroup of LCS developed according the same mechanism involved in Merkel cell carcinoma pathogenesis. We proposed LCH developed from an inflammatory process that was sustained due to gene mutations. We hypothesized that MCPyV infection triggered an IL-1 activation loop that lies beneath the pathogenesis of LCH and propose a new triple-factor model.

Abstract Objective We built India Allele Finder, an online searchable database and command line tool, that gives researchers access to variant frequencies of Indian Telugu individuals, using publicly available fastq data from the 1000 Genomes Project. Access to appropriate population-based genomic variant annotation can accelerate the interpretation of genomic sequencing data. In particular, exome analysis of individuals of Indian descent will identify population variants not reflected in European exomes, complicating genomic analysis for such individuals. Results India Allele Finder offers improved ease-of-use to investigators seeking to identify and annotate sequencing data from Indian populations. We describe the use of India Allele Finder to identify common population variants in a disease quartet whole exome dataset, reducing the number of candidate single nucleotide variants from 84 to 7. India Allele Finder is freely available to investigators to annotate genomic sequencing data from Indian populations. Use of India Allele Finder allows efficient identification of population variants in genomic sequencing data, and is an example of a population-specific annotation tool that simplifies analysis and encourages international collaboration in genomics research.

The originality of time optimization afforded by the use of mobile telephones is due to the fact that, not only is time condensed through better organization in the sequencing and processing of tasks, but also through the doubling of time with the simultaneous superposition of media time and physical time. Since its identification ten years ago, this phenomenon has continued to develop and has brought with it shortened delays, acceleration of time and the multiplication of emergency situations. However, we also witness the parallel development of reactions to this dispersion and getting lost, to too much randomness and to continually renewed ephemerality in the present. It finds its source in a logic of critical action which refuses dispossession of one’s own temporality and rhythms for generalized synchonization, and which reintroduces the thickness of mature time, reflection and meditation to counteract pressures for immediate, urgent reactions by impulse.

In this paper, a method based on the inherent event-based sampling capability of laser optical feedback interferometry (OFI) is proposed to assess the optical feedback factor C when the laser operates in the moderate and strong feedback regimes. Most of the phase unwrapping open-loop OFI algorithms rely on the estimation of C to retrieve the displacement with nanometric precision. Here, the proposed method operates in open-loop configuration and relies only on OFI’s fringe detection, thereby improving its robustness and ease of use. The proposed method is able to estimate C with a precision of C on the fly even when C is varying due to speckle. To the best of the authors’ knowledge, these are the first reported results of time-varying C estimation. In addition, through C estimation over time, it could pave the way not only to higher performance phase unwrapping algorithms but also to a better control of the optical feedback level via the use of an adaptive lens and thus to better displacement retrieval performances.

At the micrometric scale, vessels or skin capillaries network architecture can provide useful information for human health management. In this paper, from simulation to in vitro, we investigate some limits and interests of optical feedback interferometry (OFI) for blood flow imaging of skin vascularization. In order to analyze the tissue scattering effect on OFI performances, a series of skin-tissue simulating optical phantoms have been designed, fabricated and characterized. The horizontal (2D) and vertical (depth penetration) sensing resolution of the OFI sensor have been estimated. The experimental results that we present on this study are showing a very good accordance with theoretical models. In the case of a skin phantom of 0.5 mm depth with a scattering coefficient from 0 to 10.8 mm−1, the presented OFI system is able to distinguish a pair of micro fluidic channels (100 µm × 100 µm) spaced by 10 µm. Eventually, an in vivo test on human skin is presented and, for the first time using an OFI sensor, a 2D blood flow image of a vein located just beneath the skin is computed.

Every living cell is covered with a dense and complex layer of glycans on the cell surface, which have important functions in the interaction between cells and their environment. Glycosphingolipids (GSLs) are glycans linked to lipid molecules that together with sphingolipids, sterols, and proteins form plasma membrane lipid rafts that contribute to membrane integrity and provide specific recognition sites. GSLs are subdivided into three major series (globo-, ganglio-, and neolacto-series) and are synthesized in a non-template driven process by enzymes localized in the ER and Golgi apparatus. Altered glycosylation of lipids are known to be involved in tumor development and metastasis. Metastasis is frequently linked with reversible epithelial-to-mesenchymal transition (EMT), a process involved in tumor progression, and the formation of new distant metastatic sites (mesenchymal-to-epithelial transition or MET). On a single cell basis, cancer cells lose their epithelial features to gain mesenchymal characteristics via mechanisms influenced by the composition of the GSLs on the cell surface. Here, we summarize the literature on GSLs in the context of reversible and cancer-associated EMT and discuss how the modification of GSLs at the cell surface may promote this process.

The relationship between ABO blood groups (BG) and risk of incidence in cancers including gynecological cancers has been widely studied, showing increased incidence risk for BG A patients. As available data are inconsistent we investigated whether BG and their anti-glycan antibodies (anti-A and anti-B) have prognostic values in gynecological cancers. We retrospectively evaluated 974 patients with gynecological cancers in three cancer centers (Switzerland and Australia) between 1974 and 2014 regarding the relationships between clinico-pathological findings and the BG. Time to disease recurrence was significantly influenced by BG in patients with ovarian (n = 282) and vulvar (n = 67) cancer. BG O or B patients showed a significantly increased risk for ovarian cancer relapse compared to A, 59% and 82%, respectively (p = 0.045; HR O vs A = 1.59 (CI 1.01-2.51) and (p = 0.036; HR A vs B = 0.55 (CI 0.32-0.96). Median time to relapse for advanced stage (n = 126) ovarian cancer patients was 18.2 months for BG O and 32.2 for A (p = 0.031; HR O vs A = 2.07 (CI 1.07-4.02)). BG also significantly influenced relapse-free survival in patients with vulvar cancer (p = 0.002), with BG O tending to have increased relapse risk compared to A (p = 0.089). Blood groups hence associate with recurrence in ovarian and vulvar cancer: women with BG O seem to have a lower ovarian cancer incidence, however are more likely to relapse earlier. The significance of the BG status as a prognostic value is evident and may be helpful to oncologists in prognosticating disease outcome and selecting the appropriate therapy.

Travel surveys can collect information about practices and behaviours of individual mobility. These surveys and the resulting analyses are necessary for the development and evaluation of public transport policies. Usually, these surveys use traditional methods for data collection with standard survey methods. Today, new survey protocols and data collection methods emerge thanks to the potential of new technologies. Geo-localization technologies, such as GPS, can be an interesting support for new types of so-called "assisted" surveys. However, these technologies may provide new types of challenges in the context of surveys with representative samples of the population. Furthermore, the use of location technologies requires modifying significantly the survey protocols. To identify the potential benefit of geo-localization technologies for travel surveys, we conducted a comparative study between a classic travel survey (Household Travel Survey) and a GPS assisted travel survey. This comparative evaluation also allowed us to identify the main technological and methodological limitations of GPS assisted travel surveys.

The sociological study of youth religion is a growing field of research. This essay focuses on the rising number of young Korean adult believers from five downtown religious congregations in Seoul who are leaving their congregations due to increased cynicism and as they re-examine their religious faith. Based on a series of in-depth interviews (Focus Group Interviews (FGIs)) conducted with both young adult believers and priests, this paper examines several key factors including why young adult believers decide to leave their congregations, how their congregations react to the changing environment, and whether they can develop innovative approaches to deal with the emerging situation. Although each and every one of the congregations considers this phenomenon to be a serious issue, the solutions they have applied to retain and attract young adult believers in and to their faith have so far been ineffective. Nonetheless, it is likely that the reason behind the failure to keep more young adults in their faith and congregation may correlate to how competitive and innovative solutions provided by the congregational leadership are as they attempt to tackle the ongoing issue of retaining and attracting young adult congregational members.

Altered levels of naturally occurring anti-glycan antibodies (AGA) circulating in human blood plasma are found in different pathologies including cancer. Here the levels of AGA directed against 22 negatively charged (sialylated and sulfated) glycans were assessed in high-grade serous ovarian cancer (HGSOC, n = 22) patients and benign controls (n = 31) using our previously developed suspension glycan array (SGA). Specifically, the ability of AGA to differentiate between controls and HGSOC, the most common and aggressive type of ovarian cancer with a poor outcome was determined. Results were compared to CA125, the commonly used ovarian cancer biomarker. AGA to seven glycans that significantly (P

Glycans are important partners in many biological processes, including carcinogenesis. The rapidly developing field of functional glycomics becomes one of the frontiers of biology and biomedicine. Aberrant glycosylation of proteins and lipids occurs commonly during malignant transformation and leads to the expression of specific tumor-associated glycans. The appearance of aberrant glycans on carcinoma cells is typically associated with grade, invasion, metastasis and overall poor prognosis. Cancer-associated carbohydrates are mostly located on the surface of cancer cells and are therefore potential diagnostic biomarkers. Currently, there is increasing interest in cancer-associated aberrant glycosylation, with growing numbers of characteristic cancer targets being detected every day. Breast and ovarian cancer are the most common and lethal malignancies in women, respectively, and potential glycan biomarkers hold promise for early detection and targeted therapies. However, the acceleration of research and comprehensive multi-target investigation of cancer-specific glycans could only be successfully achieved with the help of a combination of novel high-throughput glycomic approaches.

The past 40 years have seen significant improvements in both event-free and overall survival for children with acute lymphoblastic or acute myeloid leukemia (ALL or AML respectively). Serial national and international clinical trials have optimised the use of conventional chemotherapeutic drugs and, along with improvements in supportive care that have enabled the delivery of more intensive regimens, have been responsible for the major improvements in patient outcome seen over the past few decades,. The benefits of dose intensification, however, have likely now been maximised, and over the same period, the identification of new cytotoxic drugs has been limited.As the biology of leukemogenesis has become better understood, key molecules and intracellular pathways have been identified that offer the possibility of targeting directly the leukemia cells whilst sparing normal cells. Consequently, there is now a drive to develop novel leukemia-specific or ‘targeted’ therapies. These new classes of drugs will have mechanisms of action, toxicities and therapeutic indices quite different from conventional cytotoxic drugs previously encountered, thus rendering current clinical trial methodologies inappropriate; clinical trial methods will need to be adapted to accommodate these features of these new classes of drugs. This review will address the challenges and some of the techniques for developing clinical trials for targeted therapies.

The association between pyogenic liver abscesses and schistosomiasis has been confirmed by clinical and experimental studies. In this retrospective study of 78 patients with pyogenic liver abscesses the association with schistosomiasis has been investigated. Pyodermitis, a known focus of bacteremia, was observed in 19 patients (24%). Blood eosinophilia was observed in 30 patients (39%). Staphylococcus aureus was cultured from abscesses in 17 out of 38 patients (45%). Forty-one out of 57 patients (53%) had stool examination. Schistosoma mansoni was the main parasite identified. Eggs of S. mansoni were also identified in liver biopsies in 7 out of 19 patients who did the exam. The large number of young patients with liver abscesses described here is different from what has been observed in developed countries. This clinical study provide support for the concept that granulomas of S. mansoni in the liver are foci for colonization with S. aureus, which in presence of staphylococcal bacteremia can form liver abscesses.

Reverse Transcription - quantitative Polymerase Chain Reaction (RT-qPCR) is a standard technique in most laboratories. The selection of reference genes is essential for data normalization and the selection of suitable reference genes remains critical. Our aim was to 1) review the literature since implementation of the MIQE guidelines in order to identify the degree of acceptance; 2) compare various algorithms in their expression stability; 3) identify a set of suitable and most reliable reference genes for a variety of human cancer cell lines. A PubMed database review was performed and publications since 2009 were selected. Twelve putative reference genes were profiled in normal and various cancer cell lines (n = 25) using 2-step RT-qPCR. Investigated reference genes were ranked according to their expression stability by five algorithms (geNorm, Normfinder, BestKeeper, comparative ΔCt, and RefFinder). Our review revealed 37 publications, with two thirds patient samples and one third cell lines. qPCR efficiency was given in 68.4% of all publications, but only 28.9% of all studies provided RNA/cDNA amount and standard curves. GeNorm and Normfinder algorithms were used in 60.5% in combination. In our selection of 25 cancer cell lines, we identified HSPCB, RRN18S, and RPS13 as the most stable expressed reference genes. In the subset of ovarian cancer cell lines, the reference genes were PPIA, RPS13 and SDHA, clearly demonstrating the necessity to select genes depending on the research focus. Moreover, a cohort of at least three suitable reference genes needs to be established in advance to the experiments, according to the guidelines. For establishing a set of reference genes for gene normalization we recommend the use of ideally three reference genes selected by at least three stability algorithms. The unfortunate lack of compliance to the MIQE guidelines reflects that these need to be further established in the research community.