An automated analysis of highly complex flow cytometry-based proteomic data.

The combination of color-coded microspheres as carriers and flow cytometry as a detection platform provides new opportunities for multiplexed measurement of biomolecules. Here, we developed a software tool capable of automated gating of color-coded microspheres, automatic extraction of statistics from all subsets and validation, normalization, and cross-sample analysis. The approach presented in this article enabled us to harness the power of high-content cellular proteomics. In size exclusion chromatography-resolved microsphere-based affinity proteomics (Size-MAP), antibody-coupled microspheres are used to measure biotinylated proteins that have been separated by size exclusion chromatography. The captured proteins are labeled with streptavidin phycoerythrin and detected by multicolor flow cytometry. When the results from multiple size exclusion chromatography fractions are combined, binding is detected as discrete reactivity peaks (entities). The information obtained might be approximated to a multiplexed western blot. We used a microsphere set with >1,000 subsets, presenting an approach to extract biologically relevant information. The R-project environment was used to sequentially recognize subsets in two-dimensional space and gate them. The aim was to extract the median streptavidin phycoerythrin fluorescence intensity for all 1,000+ microsphere subsets from a series of 96 measured samples. The resulting text files were subjected to algorithms that identified entities across the 24 fractions. Thus, the original 24 data points for each antibody were compressed to 1-4 integrated values representing the areas of individual antibody reactivity peaks. Finally, we provide experimental data on cellular protein changes induced by treatment of leukemia cells with imatinib mesylate. The approach presented here exemplifies how large-scale flow cytometry data analysis can be efficiently processed to employ flow cytometry as a high-content proteomics method.

Small left atrium and mild mitral regurgitation predict super-response to cardiac resynchronization therapy.

AIMS: Cardiac resynchronization therapy (CRT) can result in profound reverse remodelling. The goal of this study was to identify factors predictive of such beneficial response. METHODS AND RESULTS: Super-response to CRT was defined as normalization or near normalization of left ventricular systolic function without recognized reversible causes of heart failure. In a retrospective study, we compared baseline demographic, electrocardiogram, and echocardiographic characteristics of super-responders (n = 21) with a population of unselected consecutive cardiac CRT patients (Control 1, n = 330) and another sex-, age-, and aetiology-matched control group (Control 2, n = 43). Compared with Control 1, super-responders had significantly smaller left ventricular end-diastolic diameter (65.4 ± 6.4 vs. 73.4 ± 9.3 mm, P = 0.0001), higher ejection fraction (0.25 ± 0.05 vs. 0.22 ± 0.04, P = 0.004), smaller degree of mitral regurgitation (MR; mean value 1.9 ± 0.9 vs. 2.6 ± 0.8, P = <0.0001), and smaller left atrium (LA; 42.8 ± 4.6 vs. 50.0 ± 6.5 mm, P < 0.0001). Septal flash and inter-ventricular mechanical dyssynchrony were both more frequent among super-responders than in Control 2 subjects (93.8 vs. 69.8%; P = 0.01, and 93.8 vs. 62.8%; P = 0.01, respectively). In a multivariate analysis, smaller LA diameter and milder MR remained independent predictors of super-response. CONCLUSION: Super-response to cardiac CRT was associated with less advanced left-sided structural involvement as described by echocardiography. In particular, smaller LA and milder MR were independent predictors of pronounced reverse remodelling.

Complications post simple exodontia: A systematic review.

Exodontia procedures are not without complications, which are the dentist's responsibility to avoid by taking into account clinical, imaging, systemic, and operative factors, among others. The purpose of this systematic review is to determine and analyze the prevalence of complications post simple exodontia (CPES). The method used in this systematic review was adapted from the Cochrane Handbook and PRISMA statement. A systematic search was conducted in PubMed, Scopus and ScienceDirect using the search terms "Exodontia" AND "Complications". The search was conducted from the starting coverage date to January 31, 2020. The inclusion criteria were studies on simple exodontia, studies on CPES prevalence and human studies. Studies on complications after third molar exodontia, generalities in exodontia, narratives and systematics literature reviews, book chapters, and animal studies were excluded. A total of 1,446 articles were found in the first search using the search strategy (725 in PubMed, 96 in Scopus and 631 in ScienceDirect). After duplicates were removed, 948 articles were obtained. After reading the title and abstract, 9 articles were read in full. Finally, 3 articles were included in the review, with the most common complications being trismus, alveolitis, pain, dehiscence, infections, and retained roots. Trismus of the chewing muscles, alveolitis and retained roots were the most prevalent CPES, which were most likely related to the surgeon's experience, surgery duration and tissue trauma during surgery.

Feasibility, safety, and early outcomes of direct carotid artery stent implantation with use of the FilterWire EZ Embolic Protection System.

OBJECTIVES: To report on early outcomes of prospective single-center registry, which evaluated feasibility, safety, and effectiveness of direct carotid stenting using FilterWire EZ Embolic Protection System in high-risk patients. BACKGROUND: Little is known about the use of direct carotid stenting with FilterWire EZ Embolic Protection System and different carotid stents. METHODS: Between September 2005 and August 2008, 176 consecutive patients (70 +/- 9 years, 36% symptomatic, 89% at high risk) underwent 214 direct carotid artery stenting procedures with FilterWire EZ Embolic Protection System and different carotid stents. This study included both symptomatic (>or=50% carotid artery stenosis) as well as asymptomatic (>or=70% carotid artery stenosis) patients. The primary endpoint was the rate of 30-day adverse events, defined as neurological death, stroke, or myocardial infarction. RESULTS: We implanted 218 stents (55% with closed cell design) in 214 procedures. The technical success rate of direct stenting was 96%. Carotid artery stenosis before and after direct carotid artery stenting were 82% +/- 10% and 10% +/- 13%, respectively. The median of fluoroscopic time was 6 min (range 2.5-31.5 min). There were two (0.9%) minor strokes and one (0.5%) myocardial infarction within 30 post-procedural days; the primary endpoint occurred in three patients (1.4%). Two patients expired from nonneurological causes unrelated to carotid stenting. CONCLUSION: Direct carotid stenting with FilterWire EZ Embolic Protection System is feasible and safe. Short-term results with respect to neurological events are acceptable.

Carotid artery stenting in asymptomatic and surgically high-risk patients: single-centre, single-operator results.

BACKGROUND: Stroke represents the third leading cause of death in developed countries and the leading cause of disability in the elderly. Because asymptomatic, surgically high-risk patients have been systematically excluded from randomized trials of carotid endarterectomy and medical therapy, the management of this group of patients is still controversial. A single-centre, single-operator registry was analyzed to evaluate feasibility and safety of carotid artery stenting (CAS) with distal protection devices in consecutive, asymptomatic, surgically high-risk patients who were scheduled for endovascular treatment of significant carotid stenoses. METHODS: A total of 122 consecutive, surgically high-risk, asymptomatic patients (150 carotid arteries, 59% men, mean [± SD] age 69±9 years) with severe carotid stenosis and one or more high-risk features for carotid endarterectomy were scheduled for CAS. All procedures were performed in a single centre by a single operator. All patients were prospectively asked to undergo a clinical 30-day follow-up. RESULTS: A total of 154 stents were implanted in 150 carotid arteries. The primary success rate was 98.7%. The rates of stenosis before and after direct CAS were 81%±9% and 10%±13%, respectively. The median of fluoroscopic time of direct CAS was 6 min (range 2.5 min to 31.5 min). At 30 days, data were available in all patients. The combined 30-day mortality and stroke rate was 1.3%. CONCLUSIONS: Short-term outcomes of CAS in asymptomatic, surgically high-risk patients treated by a single operator suggest a low periprocedural complication rate.

Thirty-day outcomes of direct carotid artery stenting with cerebral protection in high-risk patients.

BACKGROUND: Implantation of a carotid artery stent after predilation is a standard approach in the endovascular treatment of carotid artery stenoses. Stenting without predilation may be an alternative approach in a certain subset of patients. The present prospective, single-center registry was designed to evaluate the feasibility and safety of direct carotid artery stenting (DCAS) in high-risk patients. METHODS AND RESULTS: Symptomatic patients with stenosis >50% and asymptomatic patients with stenosis >70% were eligible for enrolment. Criteria for high-risk patients included: need or history of open heart surgery, history of myocardial infarction, multivessel coronary artery disease, left ventricular dysfunction (ejection fraction < or =40%), severe pulmonary or renal disease, significant contralateral carotid disease, previous endarterectomy, and age > or =80 years. All procedures were performed using a filter protection device. Patients underwent complete clinical examination before and after DCAS and at 30-day follow-up. A total of 83 consecutive patients (45 males, 68+/-9 years, 33% symptomatic) underwent 100 procedures and 103 stents were deployed successfully. The technical success rate of stenting was 100%. Predilation of carotid stenosis was necessary in 1 (1%) procedure. Carotid-artery stenoses before and after DCAS were 80+/-9% and 7+/-9%, respectively. The median fluoroscopic time for DCAS was 7 min. The overall rate of in-hospital major adverse cerebrovascular events (death, stroke, myocardial infarction) was 5% (2 minor strokes, 3 transient attacks). There was 1 (1%) minor stroke within the 30-day follow-up. CONCLUSION: DCAS is feasible and can be performed with an acceptable risk in high-risk patients.