Efectos de la exposición pasiva al aerosol de los cigarrillos electrónicos: una revisión de tema / Effects of passive exposure to aerosol from electronic cigarettes: a review

Resumen Los cigarrillos electrónicos sustituyeron el tabaco del cigarrillo convencional por un e-liquid compuesto por varios compuestos orgánicos; estos entraron al mercado sin mayores pruebas toxicológicas preclínicas o ensayos de seguridad a nivel mundial, generando un gran número de personas expuestas al aerosol de segunda mano, en quienes los posibles riesgos aún no han sido dilucidados. El objetivo de esta revisión es identificar los riesgos para la salud de personas expuestas al aerosol de segunda mano de cigarrillos electrónicos. La búsqueda bibliográfica realizó una revisión en las bases de datos PubMed, Scielo y EBSCO, incluyendo estudios realizados en humanos, animales e in vitro. Los principales hallazgos fueron exacerbaciones de asma, enfermedad pulmonar obstructiva crónica, efectos proinflamatorios, estrés oxidativo y ansiedad. La evidencia encontró efectos adversos en personas expuestas al aerosol de segunda mano del cigarrillo electrónico; se destacan exacerbaciones de asma, neumonitis por hipersensibilidad, inflamación y estrés oxidativo.

Abstract Electronic cigarettes replaced the tobacco leaf in conventional cigarettes with an e-liquid composed of multiple organic compounds; these entered the market without major preclinical toxicological tests or safety trials worldwide. Generating a large number of people exposed to second-hand aerosol, to whom the possible risks have not yet been elucidated. The objective of this review was to identify the health risks of people exposed to second-hand aerosol from electronic cigarettes. The review was carried out using PubMed, Scielo and EBSCO databases, including studies carried out in humans, animals and invitro. The main findings were exacerbations of asthma and chronic obstructive pulmonary disease, pro-inflammatory effects, oxidative stress and anxiety. Evidence found adverse effects in people exposed to second-hand aerosol from electronic cigarettes; highlighting exacerbations of asthma, hypersensitivity pneumonitis, inflammation and oxidative stress.

Erratum: Reduced expression of enolase-1 correlates with high intracellular glucose levels and increased senescence in cisplatin-resistant ovarian cancer cells.

[This corrects the article on p. 1275 in vol. 12, PMID: 32355541.].

Reduced expression of enolase-1 correlates with high intracellular glucose levels and increased senescence in cisplatin-resistant ovarian cancer cells.

Despite good responses to first-line treatment with platinum-based combination chemotherapy, most ovarian cancer patients will relapse and eventually develop a platinum-resistant disease with a poor overall prognosis. The molecular events leading to the cisplatin resistance of ovarian cancer cells are not fully understood. Here, we performed a proteomic analysis to identify protein candidates deregulated in a cisplatin-resistant ovarian cancer cell line (A2780CP20) in comparison to their sensitive counterpart (A2780). Forty-eight proteins were differentially abundant in A2780CP20, as compared with A2780, cells. Enolase-1 (ENO1) was significantly decreased in cisplatin-resistant ovarian cancer cells. Western blots and RT-PCR confirmed our findings. Ectopic ENO1 expression increased the sensitivity of ovarian cancer cells to cisplatin treatment. In contrast, small-interfering (siRNA)-based ENO1 silencing in A2780 cells reduced the sensitivity of these cells to cisplatin treatment. Whereas glucose consumption was lower, intracellular levels were higher in cisplatin-resistant ovarian cancer cells as compared with their cisplatin-sensitive counterparts. Senescence-associated ß-galactosidase (ß-Gal) levels were higher in cisplatin-resistant ovarian cancer cells as compared with cisplatin-sensitive ovarian cancer cells. ß-Gal levels were decreased in ENO1 overexpressed clones. Protein levels of the cell cycle regulators and senescence markers p21 and p53 showed opposite expression patterns in cisplatin-resistant compared with cisplatin sensitive cells. Our studies suggest that decreased expression of ENO1 promotes glucose accumulation, induces senescence, and leads to cisplatin resistance of ovarian cancer cells.

NTRK fusion detection across multiple assays and 33,997 cases: diagnostic implications and pitfalls.

With the FDA approval of larotrectinib, NTRK fusion assessment has recently become a standard part of management for patients with locally advanced or metastatic cancers. Unlike somatic mutation assessment, the detection of NTRK fusions is not straightforward, and various assays exist at the DNA, RNA, and protein level. Here, we investigate the performance of immunohistochemistry and DNA-based next-generation sequencing to indirectly or directly detect NTRK fusions relative to an RNA-based next-generation sequencing approach in the largest cohort of NTRK fusion positive solid tumors to date. A retrospective analysis of 38,095 samples from 33,997 patients sequenced by a targeted DNA-based next-generation sequencing panel (MSK-IMPACT), 2189 of which were also examined by an RNA-based sequencing assay (MSK-Fusion), identified 87 patients with oncogenic NTRK1-3 fusions. All available institutional NTRK fusion positive cases were assessed by pan-Trk immunohistochemistry along with a cohort of control cases negative for NTRK fusions by next-generation sequencing. DNA-based sequencing showed an overall sensitivity and specificity of 81.1% and 99.9%, respectively, for the detection of NTRK fusions when compared to RNA-based sequencing. False negatives occurred when fusions involved breakpoints not covered by the assay. Immunohistochemistry showed overall sensitivity of 87.9% and specificity of 81.1%, with high sensitivity for NTRK1 (96%) and NTRK2 (100%) fusions and lower sensitivity for NTRK3 fusions (79%). Specificity was 100% for carcinomas of the colon, lung, thyroid, pancreas, and biliary tract. Decreased specificity was seen in breast and salivary gland carcinomas (82% and 52%, respectively), and positive staining was often seen in tumors with neural differentiation. Both sensitivity and specificity were poor in sarcomas. Selection of the appropriate assay for NTRK fusion detection therefore depends on tumor type and genes involved, as well as consideration of other factors such as available material, accessibility of various clinical assays, and whether comprehensive genomic testing is needed concurrently.

Whole slide imaging equivalency and efficiency study: experience at a large academic center.

Whole slide imaging is Food and Drug Administration-approved for primary diagnosis in the United States of America; however, relatively few pathology departments in the country have fully implemented an enterprise wide digital pathology system enabled for primary diagnosis. Digital pathology has significant potential to transform pathology practice with several published studies documenting some level of diagnostic equivalence between digital and conventional systems. However, whole slide imaging also has significant potential to disrupt pathology practice, due to the differences in efficiency of manipulating digital images vis-à-vis glass slides, and studies on the efficiency of actual digital pathology workload are lacking. Our randomized, equivalency and efficiency study aimed to replicate clinical workflow, comparing conventional microscopy to a complete digital pathology signout using whole slide images, evaluating the equivalency and efficiency of glass slide to whole slide image reporting, reflective of true pathology practice workloads in the clinical setting. All glass slides representing an entire day's routine clinical signout workload for six different anatomic pathology subspecialties at Memorial Sloan Kettering Cancer Center were scanned on Leica Aperio AT2 at ×40 (0.25 µm/pixel). Integration of whole slide images for each accessioned case is through an interface between the Leica eSlide manager database and the laboratory information system, Cerner CoPathPlus. Pathologists utilized a standard institution computer workstation and viewed whole slide images through an internally developed, vendor agnostic whole slide image viewer, named the "MSK Slide Viewer". Subspecialized pathologists first reported on glass slides from surgical pathology cases using routine clinical workflow. Glass slides were de-identified, scanned, and re-accessioned in the laboratory information system test environment. After a washout period of 13 weeks, pathologists reported the same clinical workload using whole slide image integrated within the laboratory information system. Intraobserver equivalency metrics included top-line diagnosis, margin status, lymphovascular and/or perineural invasion, pathology stage, and the need to order ancillary testing (i.e., recuts, immunohistochemistry). Turnaround time (efficiency) evaluation was defined by the start of each case when opened in the laboratory information system and when the case was completed for that day (i.e., case sent to signout queue or pending ancillary studies). Eight pathologists participated from the following subspecialties: bone and soft tissue, genitourinary, gastrointestinal, breast, gynecologic, and dermatopathology. Glass slides signouts comprised of 204 cases, encompassing 2091 glass slides; and digital signouts comprised of 199 cases, encompassing 2073 whole slide images. The median whole slide image file size was 1.54 GB; scan time/slide, 6 min 24 s; and scan area 32.1 × 18.52 mm. Overall diagnostic equivalency (e.g., top-line diagnosis) was 99.3% between digital and glass slide signout; however, signout using whole slide images showed a median overall 19% decrease in efficiency per case. No significant difference by reader, subspecialty, or specimen type was identified. Our experience is the most comprehensive study to date and shows high intraobserver whole slide image to glass slide equivalence in reporting of true clinical workflows and workloads. Efficiency needs to improve for digital pathology to gain more traction among pathologists.

Single cell metabolic profiling of tumor mimics.

Chemical cytometry employs modern analytical methods to study the differences in composition between single cells to better understand development, cellular differentiation, and disease. Metabolic cytometry is a form of chemical cytometry wherein cells are incubated with and allowed to metabolize fluorescently labeled small molecules. Capillary electrophoresis with laser-induced fluorescence detection is then used to characterize the extent of metabolism at the single cell level. To date, all metabolic cytometry experiments have used conventional two-dimensional cell cultures. HCT 116 spheroids are a three-dimensional cell culture system, morphologically and phenotypically similar to tumors. Here, intact HCT 116 multicellular spheroids were simultaneously incubated with three fluorescently labeled glycosphingolipid substrates, GM3-BODIPY-FL, GM1-BODIPY-TMR, and lactosylceramide-BODIPY-650/665. These substrates are spectrally distinct, and their use allows the simultaneous probing of metabolism at three different points in the glycolipid metabolic cascade. Beginning with intact spheroids, a serial trypsinization and trituration procedure was used to isolate single cells from spatially distinct regions of the spheroid. Cells from the distinct regions showed unique metabolic patterns. Treatment with the lysosomal inhibitor and potential chemotherapeutic chloroquine consistently decreased catabolism for all substrates. Nearly 200 cells were taken for analysis. Principal component analysis with a multivariate measure of precision was used to quantify cell-to-cell variability in glycosphingolipid metabolism as a function of cellular localization and chloroquine treatment. While cells from different regions exhibited differences in metabolism, the heterogeneity in metabolism did not differ significantly across the experimental conditions.

Manipulating ionic strength to improve single cell electrophoretic separations.

A capillary electrophoresis system with ultrasensitive two-color laser-induced fluorescence detection was used to probe the effect of ionic strength on single cell separations of glycosphingolipids. Differentiated PC12 cells were incubated with two ganglioside substrates tagged with different fluorophores within the BODIPY family such that two distinct metabolic patterns could be simultaneously monitored. Aspiration of single differentiated PC12 cells suspended in a phosphate-buffered saline solution showed excessive peak dispersion, poor resolution, and peak efficiencies below 100,000 theoretical plates. Aspiration of single differentiated PC12 cells suspended in deionized water corrected peak dispersion. Average peak efficiencies ranged between 400,000 and 600,000 theoretical plates. Improved performance was due to the dilution of the high salt concentrations inside of single neuronal-like cells to produce field amplified sample stacking. Single cell separations showed the highest resolution when aspiration of single differentiated PC12 cells suspended in deionized water were separated using a running buffer of high ionic strength. The improvement in resolution allowed for the identification of analytes not previously detected in single cell metabolism studies.