Droplet nuclei are generated during colonoscopy and are decreased by the use of carbon dioxide and water immersion technique.

OBJECTIVES: The COVID-19 pandemic has raised concerns on whether colonoscopies (CS) carry a transmission risk. The aim was to determine whether CS are aerosol-generating procedures. METHODS: This was a prospective observational trial including all patients undergoing CS at the Prince of Wales Hospital from 1 June to 31 July 2020. Three particle counters were placed 10 cm from each patient's anus and near the mouth of endoscopists and nurses. The particle counter recorded the number of particles of size 0.3, 0.5, 0.7, 1, 5, and 10 µm. Patient demographics, seniority of endoscopists, use of CO2 and water immersion technique, and air particle count (particles/cubic foot, dCF) were recorded. Multilevel modeling was used to test all the hypotheses with a post-hoc analysis. RESULTS: A total of 117 patients were recruited. During CS, the level of 5 µm and 10 µm were significantly higher than the baseline period (P = 0.002). Procedures performed by trainees had a higher level of aerosols when compared to specialists (0.3 µm, P < 0.001; 0.5 µm and 0.7 µm, P < 0.001). The use of CO2 and water immersion techniques had significantly lower aerosols generated when compared to air (CO2 : 0.3, 0.5, and 0.7 µm: P < 0.001; water immersion: 0.3 µm: P = 0.048; 0.7 µm: P = 0.03). There were no significant increases in any particle sizes during the procedure at the endoscopists' and nurses' mouth. However, 8/117 (6.83%) particle count tracings showed a simultaneous surge of all particle sizes at the patient's anus and endoscopists' and nurses' level during rectal extubation. CONCLUSION: Colonoscopy generates droplet nuclei especially during rectal extubation. The use of CO2 and water immersion techniques may mitigate these risks.

Uniform amplification of phage display libraries in monodisperse emulsions.

In this paper, we describe a complete experimental setup for the uniform amplification of libraries of phage. Uniform amplification, which multiplies every phage clone by the same amount irrespective of the growth rate of the clone is essential for phage-display screening. Amplification of phage libraries in a common solution is often non-uniform: it favors fast-growing clones and eliminates those that grow slower. This competition leads to elimination of many useful binding clones, and it is a major barrier to identification of ligands for targets with multiple binding sites such as cells, tissues, or mixtures of proteins. Uniform amplification is achieved by encapsulating individual phage clones into isolated compartments (droplets) of identical volume. Each droplet contains culture medium and an excess of host (Escherichia coli). Here, we describe microfluidics devices that generate mono-disperse droplet-based compartments, and optimal conditions for amplification of libraries of different size. We also describe the detailed synthesis of a perfluoro surfactant, which gives droplets exceptional stability. Droplets stabilized by this compound do not coalesce after many hours in shaking culture. We identified a commercially available compound (Krytox), which destabilizes these droplets to recover the amplified libraries. Overall, uniform amplification is a sequence of three simple steps: (1) encapsulation of mixture of phage and bacteria in droplets using microfluidics; (2) incubation of droplets in a shaking culture; (3) destabilization of droplets to harvest the amplified phage. We anticipate that this procedure can be easily adapted in any academic or industrial laboratory that uses phage display.

Targeted degradation of PCNA outperforms stoichiometric inhibition to result in programed cell death.

Biodegraders are targeted protein degradation constructs composed of mini-proteins/peptides linked to E3 ligase receptors. We gained deeper insights into their utility by studying Con1-SPOP, a biodegrader against proliferating cell nuclear antigen (PCNA), an oncology target. Con1-SPOP proved pharmacologically superior to its stoichiometric (non-degrading) inhibitor equivalent (Con1-SPOPmut) as it had more potent anti-proliferative effects and uniquely induced DNA damage, cell apoptosis, and necrosis. Proteomics showed that PCNA degradation gave impaired mitotic division and mitochondria dysfunction, effects not seen with the stoichiometric inhibitor. We further showed that doxycycline-induced Con1-SPOP achieved complete tumor growth inhibition in vivo. Intracellular delivery of mRNA encoding Con1-SPOP via lipid nanoparticles (LNPs) depleted endogenous PCNA within hours of application with nanomolar potency. Our results demonstrate the utility of biodegraders as biological tools and highlight target degradation as a more efficacious approach versus stoichiometric inhibition. Once in vivo delivery is optimized, biodegraders may be leveraged as an exciting therapeutic modality.

Three-Dimensional Accuracy of Digital Implant Impressions: Effects of Different Scanners and Implant Level.

PURPOSE: To compare the three-dimensional (3D) accuracy of conventional direct implant impressions with digital implant impressions from three intraoral scanners, as well as different implant levels-bone level (BL) and tissue level (TL). MATERIALS AND METHODS: Two-implant master models were used to simulate a threeunit implant-supported fixed dental prosthesis. Conventional test models were made with direct impression copings and polyether impressions. Scan bodies were hand-tightened onto master models and scanned with the three scanners. This was done for the TL and BL test groups, for a total of eight test groups (n = 5 each). A coordinate measuring machine measured linear distortions (dx, dy, dz), global linear distortion (dR), angular distortions (dθy, dθx), and absolute angular distortions (Absdθy, Absdθx) between the master models, test models, and .stl files of the digital scans. RESULTS: The mean dR ranged from 35 to 66 µm; mean dθy angular distortions ranged from -0.186 to 0.315 degrees; and mean dθx angular distortions ranged from -0.206 to 0.164 degrees. Two-way analysis of variance showed that the impression type had a significant effect on dx, dz, and Absdθy, and the implant level had a significant effect on dx and Absdθx (P < .05). Among the BL groups, the mean dR of the conventional group was lower than and significantly different from the digital test groups (P = .010), while among the TL groups, there was no statistically significant difference (P = .572). CONCLUSION: The 3D accuracy of implant impressions varied according to the impression technique and implant level. For BL test groups, the conventional impression group had significantly lower distortion than the digital impression groups. Among the digital test groups, the TR system had comparable mean linear and absolute angular distortions to the other two systems but exhibited the smallest standard deviations.

Three-dimensional accuracy of a digitally coded healing abutment implant impression system.

PURPOSE: This study examined the three-dimensional (3D) accuracy of the Encode Impression System (EN) in transferring the locations of two implants from master models to test models and compared this to the direct impression (DI) technique. The effect of interimplant angulation on the 3D accuracy of both impression techniques was also evaluated. MATERIALS AND METHODS: Seven sectional polymethyl methacrylate mandibular arch master models were fabricated with implants in the first premolar and first molar positions. The implants were placed parallel to each other or angulated mesiodistally or buccolingually with total divergent angles of 10, 20, or 30 degrees. Each master model was secured onto an aluminum block containing a gauge block, which defined the local coordinate references. Encode healing abutments were attached to the implants before impressions were made for the EN test models; pickup impression copings were attached for the DI test models. For the seven test groups of each impression technique, a total of 70 test models were fabricated (n = 5). The EN test models were sent to Biomet 3i for implant analog placement. The centroid of each implant or implant analog and the angular orientation of the long axis relative to the x- and y-axes were measured with a coordinate measuring machine. Statistical analyses were performed. RESULTS: Impression technique had a significant effect on y distortion, global linear distortion, and absolute xz and yz angular distortions. Interimplant angulation had significant effects on x and y distortions. However, neither impression technique nor interimplant angulation had a significant effect on z distortion. CONCLUSIONS: Distortions were observed with both impression techniques. However, the results suggest that EN was less accurate than DI.