CM-Dil Staining and SEC of Plasma as an Approach to Increase Sensitivity of Extracellular Nanovesicles Quantification by Bead-Assisted Flow Cytometry

The quantification of the specific disease-associated populations of circulating extracellular membrane nanovesicles (ENVs) has opened up new opportunities for liquid biopsy in cancer and other chronic diseases. However, the sensitivity of such methods is mediated by an optimal combination of the isolation and labeling approaches, and is not yet sufficient for routine clinical application. The presented study aimed to develop, characterize, and explore a new approach to non-specific ENV staining, followed by size-exclusive chromatography (SEC), which allows us to increase the sensitivity of bead-assisted flow cytometry. Plasma from healthy donors was purified from large components, stained with lipophilic CM-Dil dye, and fractionated by means of SEC. The obtained fractions were analyzed in terms of particle size and concentration using NTA, as well as vesicular markers and plasma protein content via dot-blotting. We characterized the process of CM-Dil-stained plasma fractionation in detail and indicated the fractions with optimal characteristics. Finally, we explored the sensitivity of on-bead flow cytometry for the analysis of specific populations of plasma ENVs and demonstrated the advantages and limitations of the proposed technique.

Augmented Reality in Dentistry: Uses and Applications in the Digital Era

Introduction: With all the advancements that technology has reached, Dentistry can't be left behind. In the past few years, researchers have focused on emerging technologies like Virtual and Augmented Reality with clinical practice. Objectives: This literature review aims to provide an update on the latest technological applications and development in augmented reality in the dental field. Methods: The PubMed database was reviewed, and the studies that fulfilled the inclusion criteria in the last 20 years, from 2000 to 5 May 2020, were included. Results: The search results revealed a total of 72 articles, 32 were excluded, while 40 articles were included. It’s been observed that augmented reality application is still under testing, as certain drawbacks still tie the spread of this technology in the dental field. Multiple studies have resulted in a system that is suitable for clinical use. Yet no routine clinical application has been reported. Conclusion: The research department has already covered more advanced technologies like mixed reality. Therefore, a question arises, whether augmented realty will continue to grow independently or will mixed reality dominate the field.

Augmented Reality in Dentistry: Uses and Applications in the Digital Era

Introduction: With all the advancements that technology has reached, Dentistry can't be left behind. In the past few years, researchers have focused on emerging technologies like Virtual and Augmented Reality with clinical practice. Objectives: This literature review aims to provide an update on the latest technological applications and development in augmented reality in the dental field. Methods: The PubMed database was reviewed, and the studies that fulfilled the inclusion criteria in the last 20 years, from 2000 to 5 May 2020, were included. Results: The search results revealed a total of 72 articles, 32 were excluded, while 40 articles were included. It’s been observed that augmented reality application is still under testing, as certain drawbacks still tie the spread of this technology in the dental field. Multiple studies have resulted in a system that is suitable for clinical use. Yet no routine clinical application has been reported. Conclusion: The research department has already covered more advanced technologies like mixed reality. Therefore, a question arises, whether augmented realty will continue to grow independently or will mixed reality dominate the field.

Use of Salmonella Bacteria in Cancer Therapy: Direct, Drug Delivery and Combination Approaches

Over the years, conventional cancer treatments, such as chemotherapy with only a limited specificity for tumors, have undergone significant improvement. Moreover, newer therapies such as immunotherapy have undergone a revolution to stimulate the innate as well as adaptive immune responses against the tumor. However, it has been found that tumors can be selectively colonized by certain bacteria, where they can proliferate, and exert direct oncolytic effects as well as stimulating the immune system. Bacterial-mediated cancer therapy (BMCT) is now one example of a hot topic in the antitumor field. Salmonella typhimurium is a Gram-negative species that generally causes self-limiting gastroenteritis in humans. This species has been designed and engineered in order to be used in cancer-targeted therapeutics. S. typhimurium can be used in combination with other treatments such as chemotherapy or radiotherapy for synergistic modification of the tumor microenvironment. Considerable benefits have been shown by using engineered attenuated strains for the diagnosis and treatment of tumors. Some of these treatment approaches have received FDA approval for early-phase clinical trials. This review summarizes the use of Salmonella bacteria for cancer therapy, which could pave the way towards routine clinical application. The benefits of this therapy include an automatic self-targeting ability, and the possibility of genetic manipulation to produce newly engineered attenuated strains. Nevertheless, Salmonella-mediated anticancer therapy has not yet been clinically established, and requires more research before its use in cancer treatment.

Is a timely assessment of the hematocrit necessary for cardiovascular magnetic resonance–derived extracellular volume measurements?

Background Cardiovascular magnetic resonance (CMR)–derived extracellular volume (ECV) requires a hematocrit (Hct) to correct contrast volume distributions in blood. However, the timely assessment of Hct can be challenging and has limited the routine clinical application of ECV. The goal of the present study was to evaluate whether ECV measurements lead to significant error if a venous Hct was unavailable on the day of CMR. Methods 109 patients with CMR T1 mapping and two venous Hcts (Hct0: a Hct from the day of CMR, and Hct1: a Hct from a different day) were retrospectively identified. A synthetic Hct (Hctsyn) derived from native blood T1 was also assessed. The study used two different ECV methods, (1) a conventional method in which ECV was estimated from native and postcontrast T1 maps using a region-based method, and (2) an inline method in which ECV was directly measured from inline ECV mapping. ECVs measured with Hct0, Hct1, and Hctsyn were compared for each method, and the reference ECV (ECV0) was defined using the Hct0. The error between synthetic (ECVsyn) and ECV0was analyzed for the two ECV methods. Results ECV measured using Hct1 and Hctsyn were significantly correlated with ECV0 for each method. No significant differences were observed between ECV0 and ECV measured with Hct1 (ECV1; 28.4 ± 6.6% vs. 28.3 ± 6.1%, p = 0.789) and between ECV0 and ECV calculated with Hctsyn (ECVsyn; 28.4 ± 6.6% vs. 28.2 ± 6.2%, p = 0.45) using the conventional method. Similarly, ECV0 was not significantly different from ECV1 (28.5 ± 6.7% vs. 28.5 ± 6.2, p = 0.801) and ECVsyn (28.5 ± 6.7% vs. 28.4 ± 6.0, p = 0.974) using inline method. ECVsyn values revealed relatively large discrepancies in patients with lower Hcts compared with those with higher Hcts. Conclusions Venous Hcts measured on a different day from that of the CMR examination can still be used to measure ECV. ECVsyn can provide an alternative method to quantify ECV without needing a blood sample, but significant ECV errors occur in patients with severe anemia.

Clinical and laboratory aspects of 3,3′,5′-triiodothyronine (reverse T3)

Reverse T3 (3,3′,5′-triiodothyronine or rT3) is the third most abundant iodothyronine circulating in human blood and is produced by the inner ring deiodination of the pro-hormone thyroxine (T4). Unlike the more abundant and active metabolite T3, the measurement of serum rT3 is yet to find a routine clinical application. As rT3 binds weakly to the T3 thyroid nuclear hormone receptors, it is thought to represent an inactive end-product of thyroid hormone metabolism, diverting T4 away from T3 production. The analysis of serum rT3 has, up until recently, been measured by competitive radioimmunoassay, but these methods have been superseded by mass-spectrometric methods which are less susceptible to interference from other more abundant iodothyronines. Serum rT3 concentration is increased as part of the non-thyroidal illness syndrome, and by administration of common medications such as amiodarone which inhibit the metabolism of rT3. Serum rT3 concentration is also affected by genetic conditions that affect the iodothyronine deiodinases, as well as thyroid transporters and transport proteins. Analysis of rT3 can provide a useful diagnostic fingerprint for these conditions. rT3 has been shown to bind extra-nuclear iodothyronine receptors with a potential role in cell proliferation; however, the clinical relevance of these findings awaits further study.

Ten-Month Evaluation of the Routine Application of Patient Moving Average for Real-Time Quality Control in a Hospital Setting

Background In recent years there has been renewed interest in patient-based real-time quality control (PBRTQC) techniques. This interest has been stimulated by the availability of new optimization and validation methods. Only a limited amount of research has focused on investigating the true operational value of PBRTQC. Therefore, we have evaluated the performance and value of recently implemented patient moving average quality control (MA QC) procedures. Methods The MA QC settings and protocols were as previously described (Clin Chem Lab Med 2019;57:1329–38) and included MA QCs for 10 chemistry and 6 hematological tests, all performed on duplicate analyzer systems. All MA QC alarms that occurred during the first 10 months of routine clinical application were investigated for assay-specific alarm rate and occurrence in time. Furthermore, the causes of these MA QC alarms were investigated, and alarm relevance was determined on the basis of total allowable bias (TBa) and error (TEa) derived from biological variations. Results During the 10-month period, 202 individual MA QC alarms occurred, resulting in an overall MA QC alarm rate of 0.030% and a frequency of 4.67 per week. Most alarms were triggered by sodium MA QC. Based on all available fully executed and documented MA QC alarm work-ups, MA QC detected errors that in 26.0% of the alarms exceeded the TBa and in 13.7% the TEa. In 9.2% of the alarms, MA QC alarming triggered instant (technical) corrections. Conclusions Routine clinical application of MA QC is feasible with maintaining a manageable number of alarms and enabling detection of relevant analytical errors.