Targeted EV to Deliver Chemotherapy to Treat Triple-Negative Breast Cancers

Triple-negative breast cancers (TNBCs) are heterogeneous and metastatic, and targeted therapy is highly needed for TNBC treatment. Recent studies showed that extracellular vesicles (EV) have great potential to deliver therapies to treat cancers. This study aimed to develop and evaluate a natural compound, verrucarin A (Ver-A), delivered by targeted EV, to treat TNBC. First, the surface expression of epidermal growth factor receptor (EGFR) and CD47 were confirmed with immunohistochemistry (IHC) staining of patient tissue microarray, flow cytometry and Western blotting. EVs were isolated from HEK 293F culture and surface tagged with anti-EGFR/CD47 mAbs to construct mAb-EV. The flow cytometry, confocal imaging and live-animal In Vivo Imaging System (IVIS) demonstrated that mAb-EV could effectively target TNBC and deliver the drug. The drug Ver-A, with dosage-dependent high cytotoxicity to TNBC cells, was packed in mAb-EV. The anti-TNBC efficacy study showed that Ver-A blocked tumor growth in both 4T1 xenografted immunocompetent mouse models and TNBC patient-derived xenograft models with minimal side effects. This study demonstrated that the targeted mAb-EV-Ver-A had great potential to treat TNBCs.

A Design Concept and Kinematic Model for a Soft Aquatic Robot with Complex Bio-mimicking Motion

Fish mortality assessments for turbine passages are currently performed by live-animal testing with up to a hundred thousand fish per year in Germany. A propelled sensor device could act as a fish surrogate. In this context, the study presented here investigates the state of the art via a thorough literature review on propulsion systems for aquatic robots. An evaluation of propulsion performance, weight, size and complexity of the motion achievable allows for the selection of an optimal concept for such a fish mimicking device carrying the sensors. In the second step, the design of a bioinspired soft robotic fish driven by an unconventional drive system is described. It is based on piezoceramic actuators, which allow for motion with five degrees of freedom (DOF) and the creation of complex bio-mimicking body motions. A kinematic model for the motion’s characteristics is developed, to achieve accurate position feedback with the use of strain gauges. Optical measurements validate the complex deformation of the body and deliver the basis for the calibration of the kinematic model. Finally, it can be shown, that the calibrated model presented allows the tracking of the deformation of the entire body with an accuracy of 0.1 mm.

Genetic Parameters for Growth, Ultrasound and Carcass Traits in New Zealand Beef Cattle and Their Correlations with Maternal Performance

Research has shown that enhancing finishing performance in beef cows is feasible; however, any adverse impact of selection strategies for finishing performance on the performance of the maternal herd should be taken into account. The aim of this research was to examine the inheritance of growth, ultrasound and carcass traits in finishing beef cattle and to evaluate their correlations with maternal performance traits. Data were collected from a nationwide progeny test on commercial New Zealand hill country farms comprising a total of 4473 beef cows and their progeny. Most finishing traits were moderately to highly heritable (0.28–0.58) with the exception of meat or fat colour and ossification (0.00–0.12). Ultrasound scan traits had high genetic correlations with corresponding traits measured at slaughter (rg = 0.53–0.95) and may be used as a selection tool for improved genetic merit of the beef carcass. Fat content determined via ultrasound scanning in the live animal or at slaughter in finishing cattle is positively genetically correlated with rebreeding performance (rg = 0.22–0.39) in female herd replacements and negatively correlated with mature cow live weight (rg = −0.40 to −0.19). Low-magnitude associations were observed between the genetic merit for carcass fat traits with body condition in mature cows.

Post-Traumatic Osteoarthritis Assessment in Emerging and Advanced Pre-Clinical Meniscus Repair Strategies: A Review

Surgical repair of meniscus injury is intended to help alleviate pain, prevent further exacerbation of the injury, restore normal knee function, and inhibit the accelerated development of post-traumatic osteoarthritis (PTOA). Meniscus injuries that are treated poorly or left untreated are reported to significantly increase the risk of PTOA in patients. Current surgical approaches for the treatment of meniscus injuries do not eliminate the risk of accelerated PTOA development. Through recent efforts by scientists to develop innovative and more effective meniscus repair strategies, the use of biologics, allografts, and scaffolds have come into the forefront in pre-clinical investigations. However, gauging the extent to which these (and other) approaches inhibit the development of PTOA in the knee joint is often overlooked, yet an important consideration for determining the overall efficacy of potential treatments. In this review, we catalog recent advancements in pre-clinical therapies for meniscus injuries and discuss the assessment methodologies that are used for gauging the success of these treatments based on their effect on PTOA severity. Methodologies include histopathological evaluation of cartilage, radiographic evaluation of the knee, analysis of knee function, and quantification of OA predictive biomarkers. Lastly, we analyze the prevalence of these methodologies using a systemic PubMed® search for original scientific journal articles published in the last 3-years. We indexed 37 meniscus repair/replacement studies conducted in live animal models. Overall, our findings show that approximately 75% of these studies have performed at least one assessment for PTOA following meniscus injury repair. Out of this, 84% studies have reported an improvement in PTOA resulting from treatment.

A steep early learning curve for Endoscopic Submucosal Dissection (ESD) in the live porcine model

Background: Endoscopic Submucosal Dissection (ESD) is a demanding procedure requiring high level of expertise. ESD training programs incorporate procedures with live animal models. This study aimed to assess the early learning curve for performing ESD on live porcine models by endoscopists without any or limited previous ESD experience. Methods: In a live porcine model ESD workshop, number of resections, completeness of the resections, en bloc resections, adverse events, tutor intervention, type of knife, ESD time and size of resected specimens were recorded. ESD speed was calculated. Results: A total of 70 procedures were carried out by 17 trainees. The percentage of complete resections, en bloc resections and ESD speed increased from the first to the latest procedures (88,2% to 100%; 76,5% to 100%; 8,6 to 31,4mm2/min, respectively). The number of procedures in which a trainee needed tutor intervention and the number of adverse events also decreased throughout the procedures (4 to 0 and 6 to 0, respectively). During the workshop, when participants changed to a different type of knife, ESD speed slightly decreased (18,5mm2/min to 17,0mm2/min) and adverse events increased again (0 to 2). Conclusions: Through successive procedures, complete resections, en bloc resections and ESD speed improve whereas adverse events decrease, supporting the role of the live porcine model in the preclinical learning phase. Changing ESD knives has a momentarily negative impact on the learning curve.

Endoscopic Submucosal Dissection (ESD) skills transfer to clinical practice after hands-on workshops: an international survey

Background Endoscopic submucosal dissection (ESD) is a complex procedure, requiring enhanced technical skills. Translation into clinical practice of ESD training programs has not been documented. Our aim was to assess ESD training pathways of endoscopists participating in dedicated workshops and its clinical impact on ESD outcomes. Methods Participants of live porcine models ESD workshops, from 2013 to 2019, were included. They were invited to complete a survey focusing in human ESD performance after training, prior skills/competencies, complete learning pathway and clinical outcomes. Results From 118 invited participants, 40 (34%) completed the questionnaire. Nineteen (47%) endoscopists performed human ESD after the workshop, predominantly male (89%). At the beginning of human ESD, endoscopists had a mean of 7,7 (SD 4,1) years of endoscopic experience and were all performing endoscopic mucosal resection (EMR) and emergency endoscopy. Before ESD practice, 100% trained with live animal models and 68% with ex vivo models. The majority started clinical ESD in the lower third of the stomach or rectum (90%), with lesions ≤30mm (89%). Each endoscopist performed a median of 19 (IQR 8-32) cumulative ESDs, over a mean of 3,9 (SD 2,0) years. Total en bloc resection rate was 92%, R0 resection rate 88%, curative resection rate 86%, whereas adverse events remained <10%. Endoscopists with >10 human ESD procedures achieve clinical competence thresholds. Conclusions Participants of ESD workshops are adequately skilled prior to clinical ESD, complying with recommendations for training and properly implementing the technique. Transfer to clinical practice, of prior ESD skills obtained in hands-on training courses was documented. Structured training programs achieve clinical outcomes exceeding established standards, namely in the very initial clinical phase.

A Low-Cost, Autonomous Gait Detection and Estimation System for Analyzing Gait Impairments in Mice

With the advancement in imaging technology, many commercial systems have been developed for performing motion analysis in mice. However, available commercial systems are expensive and use proprietary software. In this paper, we describe a low-cost, camera-based design of an autonomous gait acquisition and analysis system for inspecting gait deficits in C57BL/6 mice. Our system includes video acquisition, autonomous gait-event detection, gait-parameter extraction, and result visualization. We provide a simple, user-friendly, step-by-step detailed methodology to apply well-known image processing techniques for detecting mice footfalls and calculating various gait parameters for analyzing gait abnormalities in healthy and neurotraumatic mice. The system was used in a live animal study for assessing recovery in a mouse model of Parkinson’s disease. Using the videos acquired in the study, we validate the performance of our system with receiver operating characteristic (ROC) and Hit : Miss : False (H : M : F) detection analyses. Our system correctly detected the mice footfalls with an average H : M : F score of 92.1 : 2.3 : 5.6. The values for the area under an ROC curve for all the ROC plots are above 0.95, which indicates an almost perfect detection model. The ROC and H : M : F analyses show that our system produces accurate gait detection. The results observed from the gait assessment study are in agreement with the known literature. This demonstrates the practical viability of our system as a gait analysis tool.

Feasibility of near-infrared spectroscopy for species identification and parasitological diagnosis of freshwater snails of the genus Biomphalaria (Planorbidae)

Near Infrared Spectroscopy (NIRS) has been applied in epidemiological surveillance studies of insect vectors of parasitic diseases, such as the Dengue’s mosquitoes. However, regarding mollusks, vectors of important worldwide helminth diseases such as schistosomiasis, fascioliasis and angiostrongyliasis, NIRS studies are rare. This work proposes to establish and standardize the procedure of data collection and analysis using NIRS applied to medical malacology, i.e., to mollusk vectors identifications. Biomphalaria shells and live snails were analyzed regarding several operational aspects, such as: moisture, shell side and position of the live animal for acquisition of NIR spectra. Representative spectra of Biomphalaria shells and live snails were collected using an average of 50 scans per sample and resolution of 16 cm-1. For shells, the sample should first be dried for a minimum of 15 days at an average temperature of 26±1°C, and then placed directly in the equipment measurement window with its left side facing the light beam. Live animals should be dried with absorbent paper; placed into a glass jar, and analyzed similarly to the shells. Once standardized, the technique was applied aiming at two objectives: identification of Biomphalaria using only the shells and parasitological diagnosis for Schistosoma mansoni infection. The discrimination of the three Biomphalaria species intermediate hosts of S. mansoni only by shell has technical limit due to the scarcity of organic material. Nevertheless, it was possible to differentiate B. straminea from B. tenagophila and B. glabrata with 96% accuracy. As for the parasitological diagnosis, it was possible to differentiate infected mollusks shedding S. mansoni cercariae from the non-infected ones with 82, 5% accuracy. In conclusion, the Near Infrared Spectroscopy (NIR’s) technique has proven to be an innovative and sound tool to detect infection by S. mansoni in the different species of Biomphalaria intermediate hosts.

Development and validity evidence of an objective structured assessment of technical skills score for minimally invasive linear-stapled, hand-sewn intestinal anastomoses: the A-OSATS score

Introduction The aim of this study was to develop a reliable objective structured assessment of technical skills (OSATS) score for linear-stapled, hand-sewn closure of enterotomy intestinal anastomoses (A-OSATS). Materials and methods The Delphi methodology was used to create a traditional and weighted A-OSATS score highlighting the more important steps for patient outcomes according to an international expert consensus. Minimally invasive novices, intermediates, and experts were asked to perform a minimally invasive linear-stapled intestinal anastomosis with hand-sewn closure of the enterotomy in a live animal model either laparoscopically or robot-assisted. Video recordings were scored by two blinded raters assessing intrarater and interrater reliability and discriminative abilities between novices (n = 8), intermediates (n = 24), and experts (n = 8). Results The Delphi process included 18 international experts and was successfully completed after 4 rounds. A total of 4 relevant main steps as well as 15 substeps were identified and a definition of each substep was provided. A maximum of 75 points could be reached in the unweighted A-OSATS score and 170 points in the weighted A-OSATS score respectively. A total of 41 anastomoses were evaluated. Excellent intrarater (r = 0.807–0.988, p < 0.001) and interrater (intraclass correlation coefficient = 0.923–0.924, p < 0.001) reliability was demonstrated. Both versions of the A-OSATS correlated well with the general OSATS and discriminated between novices, intermediates, and experts defined by their OSATS global rating scale. Conclusion With the weighted and unweighted A-OSATS score, we propose a new reliable standard to assess the creation of minimally invasive linear-stapled, hand-sewn anastomoses based on an international expert consensus. Validity evidence in live animal models is provided in this study. Future research should focus on assessing whether the weighted A-OSATS exceeds the predictive capabilities of patient outcomes of the unweighted A-OSATS and provide further validity evidence on using the score on different anastomotic techniques in humans.