Sialyl LewisX/A and Cytokeratin Crosstalk in Triple Negative Breast Cancer

project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. Publisher Copyright: © 2023 by the authors.Triple-negative breast cancer (TNBC) encompasses multiple entities and is generally highly aggressive and metastatic. We aimed to determine the clinical and biological relevance of Sialyl-Lewis X and A (sLeX/A)—a fucosylated glycan involved in metastasis—in TNBC. Here, we studied tissues from 50 TNBC patients, transcripts from a TNBC dataset from The Cancer Genome Atlas (TCGA) database, and a primary breast cancer cell line. All 50 TNBC tissue samples analysed expressed sLeX/A. Patients with high expression of sLeX/A had 3 years less disease-free survival than patients with lower expression. In tissue, sLeX/A negatively correlated with cytokeratins 5/6 (CK5/6, which was corroborated by the inverse correlation between fucosyltransferases and CK5/6 genes. Our observations were confirmed in vitro when inhibition of sLeX/A remarkably increased expression of CK5/6, followed by a decreased proliferation and invasion capacity. Among the reported glycoproteins bearing sLeX/A and based on the STRING tool, α6 integrin showed the highest interaction score with CK5/6. This is the first report on the sLeX/A expression in TNBC, highlighting its association with lower disease-free survival and its inverse crosstalk with CK5/6 with α6 integrin as a mediator. All in all, sLeX/A is critical for TNBC malignancy and a potential prognosis biomarker and therapeutic target.publishersversionpublishe

The 2021 WHO catalogue of Mycobacterium tuberculosis complex mutations associated with drug resistance: a genotypic analysis.

Background: Molecular diagnostics are considered the most promising route to achievement of rapid, universal drug susceptibility testing for Mycobacterium tuberculosis complex (MTBC). We aimed to generate a WHO-endorsed catalogue of mutations to serve as a global standard for interpreting molecular information for drug resistance prediction. Methods: In this systematic analysis, we used a candidate gene approach to identify mutations associated with resistance or consistent with susceptibility for 13 WHO-endorsed antituberculosis drugs. We collected existing worldwide MTBC whole-genome sequencing data and phenotypic data from academic groups and consortia, reference laboratories, public health organisations, and published literature. We categorised phenotypes as follows: methods and critical concentrations currently endorsed by WHO (category 1); critical concentrations previously endorsed by WHO for those methods (category 2); methods or critical concentrations not currently endorsed by WHO (category 3). For each mutation, we used a contingency table of binary phenotypes and presence or absence of the mutation to compute positive predictive value, and we used Fisher's exact tests to generate odds ratios and Benjamini-Hochberg corrected p values. Mutations were graded as associated with resistance if present in at least five isolates, if the odds ratio was more than 1 with a statistically significant corrected p value, and if the lower bound of the 95% CI on the positive predictive value for phenotypic resistance was greater than 25%. A series of expert rules were applied for final confidence grading of each mutation. Findings: We analysed 41 137 MTBC isolates with phenotypic and whole-genome sequencing data from 45 countries. 38 215 MTBC isolates passed quality control steps and were included in the final analysis. 15 667 associations were computed for 13 211 unique mutations linked to one or more drugs. 1149 (7·3%) of 15 667 mutations were classified as associated with phenotypic resistance and 107 (0·7%) were deemed consistent with susceptibility. For rifampicin, isoniazid, ethambutol, fluoroquinolones, and streptomycin, the mutations' pooled sensitivity was more than 80%. Specificity was over 95% for all drugs except ethionamide (91·4%), moxifloxacin (91·6%) and ethambutol (93·3%). Only two resistance mutations were identified for bedaquiline, delamanid, clofazimine, and linezolid as prevalence of phenotypic resistance was low for these drugs. Interpretation: We present the first WHO-endorsed catalogue of molecular targets for MTBC drug susceptibility testing, which is intended to provide a global standard for resistance interpretation. The existence of this catalogue should encourage the implementation of molecular diagnostics by national tuberculosis programmes. Funding: Unitaid, Wellcome Trust, UK Medical Research Council, and Bill and Melinda Gates Foundation

Multiple Scenario Generation of Subsurface Models:Consistent Integration of Information from Geophysical and Geological Data throuh Combination of Probabilistic Inverse Problem Theory and Geostatistics

Neutrinos with energies above 1017 eV are detectable with the Surface Detector Array of the Pierre Auger Observatory. The identification is efficiently performed for neutrinos of all flavors interacting in the atmosphere at large zenith angles, as well as for Earth-skimming \u3c4 neutrinos with nearly tangential trajectories relative to the Earth. No neutrino candidates were found in 3c 14.7 years of data taken up to 31 August 2018. This leads to restrictive upper bounds on their flux. The 90% C.L. single-flavor limit to the diffuse flux of ultra-high-energy neutrinos with an E\u3bd-2 spectrum in the energy range 1.0 7 1017 eV -2.5 7 1019 eV is E2 dN\u3bd/dE\u3bd < 4.4 7 10-9 GeV cm-2 s-1 sr-1, placing strong constraints on several models of neutrino production at EeV energies and on the properties of the sources of ultra-high-energy cosmic rays

Comparative study on dissolution profiles of sibutramine hydrochloride monohydrate from commercial capsules

Sibutramine hydrochloride monohydrate (SHM) has been widely used for the management of overweight and obesity. However, more restrict data have been regarded about in vitro dissolution profile of SHM from pharmaceutical dosage forms. The goal of this paper was to perform a comparative analysis on dissolution profiles of SHM from four commercial capsules (formulations F1, F2, F3 and F4) available in the Brazilian pharmaceutical market. All studied preparations reached a plateau from 85 to 100% of dissolution within 20 min in purified water, HCl 0.1 mol L-1 (pH 1.2) and phosphate buffer solution (PBS pH 6.8) that can be reported as an immediate release behavior. Formulation F4 showed the lower dissolution efficiency (73.40%) in PBS medium. However, since similarity/difference data and analysis of variance were carried out, results demonstrated no statistical differences among the evaluated formulations in the three used media. Weibull equation was chosen as the most suitable kinetic model that better adjusted the experimental dissolution data.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Validation of an analytical method for determination of Sibutramine Hydrochloride Monohydrate in capsules by Uv-Vis spectrophotometry

Las farmacopeas todavía no tienen un método oficial para la cuantificación de la sibutramina clorhidrato monohidrato (SHM). Por consiguiente un método espectrofotométrico de ultravioleta sensible, robusto y selectivo fue desarrollado y validado para la cuantificación de SHM en cápsulas. Este estudio se realizó para todos los parámetros de validación establecidos por los criterios internacionales: linealidad, precisión, exactitud y especificidad. Las medidas se hicieron por triplicado y las medias y desviaciones estándares fueron informadas. En la solución acuosa, la linealidad se obtuvo con un coeficiente de correlación de 0,9997 para el rango de concentración de 5,0 a 30,0 µg mL–1 ± La exactitud mostró los valores de 101,4 . 1,2 %, 99,1 ± 0,94 % y 102,2 ± 1,9 %, respectivamente para las concentraciones de 10,0, 20,0 y 30,0 µg mL–1 . La reproducibilidad presentó un coeficiente de variación (RSD) de 1,4359 y la repetitibilidad reveló un RSD de 1,9234. El método propuesto es simple, de bajo costo y fácil manejo para la cuantificación de SHM en cápsulas.The pharmacopeias have not provided an official method for the quantification of sibutramine hydrochloride monohydrate (SHM). Therefore a sensitive, robust and selective ultraviolet spectrophotometric method was developed and validated for the SHM quantitative determination in capsules. This study was carried out for all validation parameters established by the international guidelines. The measurements were conducted in triplicate and the mean and standard deviations were reported. Validation results on linearity, specificity, accuracy and precision were effectively performed. In the aqueous solution, the linearity was obtained with a correlation coefficient of 0.9997 for the analytical range from 5.0 to 30.0 µg mL–1 Accuracy was 101.4 ± 1.2 %, 99.1 ± 0.9 % and 102.2 ± 1.9 %, respectively for the concentrations of 10.0, 20.0 and 30.0 µg mL–1 . (The reproducibility presented a relative standard deviation (RSD . of 1.4359 and the repeatability showed RSD of 1.9234. The proposed method is a simple, low cost and easy handling approach for the SHM quantitative determination in capsules.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Validation of an analytical method for determination of Sibutramine Hydrochloride Monohydrate in capsules by Uv-Vis spectrophotometry

Las farmacopeas todavía no tienen un método oficial para la cuantificación de la sibutramina clorhidrato monohidrato (SHM). Por consiguiente un método espectrofotométrico de ultravioleta sensible, robusto y selectivo fue desarrollado y validado para la cuantificación de SHM en cápsulas. Este estudio se realizó para todos los parámetros de validación establecidos por los criterios internacionales: linealidad, precisión, exactitud y especificidad. Las medidas se hicieron por triplicado y las medias y desviaciones estándares fueron informadas. En la solución acuosa, la linealidad se obtuvo con un coeficiente de correlación de 0,9997 para el rango de concentración de 5,0 a 30,0 µg mL–1 ± La exactitud mostró los valores de 101,4 . 1,2 %, 99,1 ± 0,94 % y 102,2 ± 1,9 %, respectivamente para las concentraciones de 10,0, 20,0 y 30,0 µg mL–1 . La reproducibilidad presentó un coeficiente de variación (RSD) de 1,4359 y la repetitibilidad reveló un RSD de 1,9234. El método propuesto es simple, de bajo costo y fácil manejo para la cuantificación de SHM en cápsulas.The pharmacopeias have not provided an official method for the quantification of sibutramine hydrochloride monohydrate (SHM). Therefore a sensitive, robust and selective ultraviolet spectrophotometric method was developed and validated for the SHM quantitative determination in capsules. This study was carried out for all validation parameters established by the international guidelines. The measurements were conducted in triplicate and the mean and standard deviations were reported. Validation results on linearity, specificity, accuracy and precision were effectively performed. In the aqueous solution, the linearity was obtained with a correlation coefficient of 0.9997 for the analytical range from 5.0 to 30.0 µg mL–1 Accuracy was 101.4 ± 1.2 %, 99.1 ± 0.9 % and 102.2 ± 1.9 %, respectively for the concentrations of 10.0, 20.0 and 30.0 µg mL–1 . (The reproducibility presented a relative standard deviation (RSD . of 1.4359 and the repeatability showed RSD of 1.9234. The proposed method is a simple, low cost and easy handling approach for the SHM quantitative determination in capsules.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Validation of an analytical method for determination of Sibutramine Hydrochloride Monohydrate in capsules by Uv-Vis spectrophotometry

Las farmacopeas todavía no tienen un método oficial para la cuantificación de la sibutramina clorhidrato monohidrato (SHM). Por consiguiente un método espectrofotométrico de ultravioleta sensible, robusto y selectivo fue desarrollado y validado para la cuantificación de SHM en cápsulas. Este estudio se realizó para todos los parámetros de validación establecidos por los criterios internacionales: linealidad, precisión, exactitud y especificidad. Las medidas se hicieron por triplicado y las medias y desviaciones estándares fueron informadas. En la solución acuosa, la linealidad se obtuvo con un coeficiente de correlación de 0,9997 para el rango de concentración de 5,0 a 30,0 µg mL–1 ± La exactitud mostró los valores de 101,4 . 1,2 %, 99,1 ± 0,94 % y 102,2 ± 1,9 %, respectivamente para las concentraciones de 10,0, 20,0 y 30,0 µg mL–1 . La reproducibilidad presentó un coeficiente de variación (RSD) de 1,4359 y la repetitibilidad reveló un RSD de 1,9234. El método propuesto es simple, de bajo costo y fácil manejo para la cuantificación de SHM en cápsulas.The pharmacopeias have not provided an official method for the quantification of sibutramine hydrochloride monohydrate (SHM). Therefore a sensitive, robust and selective ultraviolet spectrophotometric method was developed and validated for the SHM quantitative determination in capsules. This study was carried out for all validation parameters established by the international guidelines. The measurements were conducted in triplicate and the mean and standard deviations were reported. Validation results on linearity, specificity, accuracy and precision were effectively performed. In the aqueous solution, the linearity was obtained with a correlation coefficient of 0.9997 for the analytical range from 5.0 to 30.0 µg mL–1 Accuracy was 101.4 ± 1.2 %, 99.1 ± 0.9 % and 102.2 ± 1.9 %, respectively for the concentrations of 10.0, 20.0 and 30.0 µg mL–1 . (The reproducibility presented a relative standard deviation (RSD . of 1.4359 and the repeatability showed RSD of 1.9234. The proposed method is a simple, low cost and easy handling approach for the SHM quantitative determination in capsules.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Validation of an Analytical Method for Determination of Sibutramine Hydrochloride Monohydrate in Capsules by Uv-Vis Spectrophotometry

SUMMARY. The pharmacopeias have not provided an official method for the quantification of sibutramine hydrochloride monohydrate (SHM). Therefore a sensitive, robust and selective ultraviolet spectrophotometric method was developed and validated for the SHM quantitative determination in capsules. This study was carried out for all validation parameters established by the international guidelines. The measurements were conducted in triplicate and the mean and standard deviations were reported. Validation results on linearity, specificity, accuracy and precision were effectively performed. In the aqueous solution, the linearity was obtained with a correlation coefficient of 0.9997 for the analytical range from 5.0 to 30.0 µg mL -1 . Accuracy was 101.4 ± 1.2 %, 99.1 ± 0.9 % and 102.2 ± 1.9 %, respectively for the concentrations of 10.0, 20.0 and 30.0 µg mL -1 . The reproducibility presented a relative standard deviation (RSD) of 1.4359 and the repeatability showed RSD of 1.9234. The proposed method is a simple, low cost and easy handling approach for the SHM quantitative determination in capsules. RESUMEN. "Validación de un Método Analítico para la Determinación de Sibutramina Clorhidrato Monohidrato en Cápsulas por Espectrofotometría UV-VIS". Las farmacopeas todavía no tienen un método oficial para la cuantificación de la sibutramina clorhidrato monohidrato (SHM). Por consiguiente un método espectrofotométri-co de ultravioleta sensible, robusto y selectivo fue desarrollado y validado para la cuantificación de SHM en cáp-sulas. Este estudio se realizó para todos los parámetros de validación establecidos por los criterios internacionales: linealidad, precisión, exactitud y especificidad. Las medidas se hicieron por triplicado y las medias y desviaciones estándares fueron informadas. En la solución acuosa, la linealidad se obtuvo con un coeficiente de correlación de 0,9997 para el rango de concentración de 5,0 a 30,0 µg mL -1 . La exactitud mostró los valores de 101,4 ± 1,2 %, 99,1 ± 0,94 % y 102,2 ± 1,9 %, respectivamente para las concentraciones de 10,0, 20,0 y 30,0 µg mL -1 . La reproducibilidad presentó un coeficiente de variación (RSD) de 1,4359 y la repetitibilidad reveló un RSD de 1,9234. El método propuesto es simple, de bajo costo y fácil manejo para la cuantificación de SHM en cápsulas