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In this study, tissue samples were stress tested to determine if freezing duration and temperature alter their mechanical properties. Tissue samples taken from the small intestine of pigs were assigned to 5 groups: fresh tissue, -28.9 °C for 7 days, -62.2 °C for 7 days, -28.9 °C for 30 days, and -62.2 °C for 30 days. Tissue was stored in PBS for the assigned temperature and duration until testing occurred with the exception of fresh tissue which was tested at sample collection. Before testing, samples were thawed in a room temperature bath, and the thickness was measured. Samples were then mounted in a biaxial test system using four anchoring rakes. Each sample was pulled to a strain of 0.2 with the corresponding forces recorded. This cycle of relaxation to 0.2 strain was repeated 5 times per sample. The thickness and force values were used to find the first Piola-Kirchhoff stress experienced at 0.2 strain and the strain energy. The average stress values in the circumferential direction were: fresh tissue: 22.3 ± 9.85 kPa; -28.9 °C for 7 days: 37.8 ± 14.1 kPa; -62.2 °C for 7 days: 46.5 ± 19.0 kPa; -28.9 °C for 30 days: 46.4 ± 22.7 kPa; -62.2 °C for 30 days: 40.1 ± 19.5 kPa. The stress and strain energy values of frozen tissue were statistically higher than the fresh tissue, although no statistical difference was found by varying duration or temperature. Based on this result, we determined that freezing tissue at any of the tested temperatures or durations increases the stiffness of the thawed tissue. This possibly occurs due to the directional formation of ice, which increases ion concentrations and glycosaminoglycan (GAG) interactions near collagen fibrils.

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With a mortality rate of 46% before the onset of COVID-19, acute respiratory distress syndrome (ARDS) affected 200,000 people in the US, causing 75,000 deaths. Mortality rates in COVID-19 ARDS patients are currently at 39%. Extrapulmonary support for ARDS aims to supplement mechanical ventilation by providing life-sustaining oxygen to the patient. A new rapid-onset, human-sized pig ARDS model in a porcine intensive care unit (ICU) was developed. The pigs were nebulized intratracheally with a high dose (4 mg/kg) of the endotoxin lipopolysaccharide (LPS) over a 2 h duration to induce rapid-onset moderate-to-severe ARDS. They were then catheterized to monitor vitals and to evaluate the therapeutic effect of oxygenated microbubble (OMB) therapy delivered by intrathoracic (IT) or intraperitoneal (IP) administration. Post-LPS administration, the PaO2 value dropped below 70 mmHg, the PaO2 /FiO2 ratio dropped below 200 mmHg, and the heart rate increased, indicating rapidly developing (within 4 h) moderate-to-severe ARDS with tachycardia. The SpO2 and PaO2 of these LPS-injured pigs did not show significant improvement after OMB administration, as they did in our previous studies of the therapy on small animal models of ARDS injury. Furthermore, pigs receiving OMB or saline infusions had slightly lower survival than their ARDS counterparts. The OMB administration did not induce a statistically significant or clinically relevant therapeutic effect in this model; instead, both saline and OMB infusion appeared to lower survival rates slightly. This result is significant because it contradicts positive results from our previous small animal studies and places a limit on the efficacy of such treatments for larger animals under more severe respiratory distress. While OMB did not prove efficacious in this rapid-onset ARDS pig model, it may retain potential as a novel therapy for the usual presentation of ARDS in humans, which develops and progresses over days to weeks.

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Biological macromolecule drugs or biologics are not suited for commonly preferred oral delivery due to their intrinsic instability and physical, chemical, or immunological barriers to the gastrointestinal tract. Ingestible capsule robots (ICR) have become a versatile platform, including use for drug delivery applications for various gastrointestinal pathologies with future potential for systemic drug delivery. In this work, a tissue attachment mechanism (TAM) for a drug delivery ICR is introduced that can facilitate a non-invasive systemic delivery of unaltered biologics via direct injection through the insensate layers of the small intestine. The main prerequisite for achieving systemic drug delivery via this device is to have a strong tissue attachment of the TAM. This study aimed to optimize the attachment success rate for drug delivery and characterize attachment duration in vivo. A fractional factorial approach was used in vivo to identify and optimize factors that most influence attachment of the TAM to maximize attachment rate. Multiple in vivo optimization levels were performed using the small intestine of anesthetized pigs, and an attachment success rate of 92% was achieved. Optimal TAMs were surgically placed in vivo to determine the duration of attachment following anesthetization and surgery recovery. The average in vivo attachment duration was 32.2±9.4 hours. This work establishes a device for consistent and reliable attachment duration, making the TAM a suitable candidate for a 24-hour systemic drug delivery platform.

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Innovative swallowable capsule technologies such as drug-loaded, dissolvable microneedles, mucoadhesive patches, and various microdevices present unique drug-carrying capabilities to overcome challenges regarding oral delivery of biologics. Here, we report a swallowable capsule for intestinal drug delivery (SCIDD) with the potential of directly injecting biological therapeutics into the insensate small intestine wall. The design, optimization, and validation of the SCIDD's primary subsystems were performed both ex-vivo and in-vivo. The assembled capsule was further tested in vivo to validate the actuation sequence and showed a 70% (n = 17) success rate in an animal model. Additionally, a drug delivery study indicated systemic uptake of adalimumab via SCIDD compared with luminal delivery in the small intestine. The pilot study presented here establishes that the novel platform could be used to orally deliver systemic biologics.

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BACKGROUND: Most complications and adverse events during laparoscopic surgery occur during initial entry into the peritoneal cavity. Among them, preperitoneal insufflation occurs when the insufflation needle is incorrectly placed, and the abdominal wall is insufflated. The objective of this study was to find a range for static pressure which is low enough to allow placement of a Veress needle into the peritoneal space without causing preperitoneal insufflation, yet high enough to separate abdominal viscera from the parietal peritoneum. METHODS: A pressure test was performed on twelve fresh porcine carcasses to determine the minimum preperitoneal insufflation pressure and the minimum initial peritoneal cavity insufflation pressure. Each porcine model had five needle placement categories. One category tested the initial peritoneal cavity insufflation pressure beneath the umbilicus. The four remaining categories tested the preperitoneal insufflation pressure at four different anatomical locations on the abdomen that can be used for initial entry. The minimum initial insufflation pressures from each carcass were then compared to the preperitoneal insufflation pressures to obtain an optimal range for initial insufflation. RESULTS: Increasing the insufflation pressure increased the probability of preperitoneal insufflation. Also, there was a statistically significant difference (p < 0.05) between the initial peritoneal cavity insufflation pressures (8.83 ± 4.19 mmHg) and the lowest preperitoneal pressures (32.54 ± 7.84 mmHg) (mean ± SD). CONCLUSION: Pressures greater than 10 mmHg resulted in initial cavity insufflation and pressures greater than 20 mmHg resulted in preperitoneal insufflation in porcine models. By knowing the minimum pressure required to separate the layers of the abdominal wall, the risk of preperitoneal insufflation can be mitigated while obtaining safe and efficient entry into the peritoneal cavity. The findings in this research are not a guideline for trocar or Veress needle placement, but instead reveal preliminary data which may lead to more studies, technology, etc.

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PURPOSE: The COVID-19 pandemic threatens our current ICU capabilities nationwide. As the number of COVID-19 positive patients across the nation continues to increase, the need for options to address ventilator shortages is inevitable. Multi-patient ventilation (MPV), in which more than one patient can use a single ventilator base unit, has been proposed as a potential solution to this problem. To our knowledge, this option has been discussed but remains untested in live patients with differing severity of lung pathology. METHODS: The objective of this study was to address ventilator shortages and patient stacking limitations by developing and validating a modified breathing circuit for two patients with differing lung compliances using simple, off-the-shelf components. A multi-patient ventilator circuit (MPVC) was simulated with a mathematical model and validated with four animal studies. Each animal study had two human-sized pigs: one healthy and one with lipopolysaccharide (LPS) induced ARDS. LPS was chosen because it lowers lung compliance similar to COVID-19. In a previous study, a control group of four pigs was given ARDS and placed on a single patient ventilation circuit (SPVC). The oxygenation of the MPVC ARDS animals was then compared to the oxygenation of the SPVC animals. RESULTS: Based on the comparisons, similar oxygenation and morbidity rates were observed between the MPVC ARDS animals and the SPVC animals. CONCLUSION: As healthcare systems worldwide deal with inundated ICUs and hospitals from pandemics, they could potentially benefit from this approach by providing more patients with respiratory care.

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In single-port access surgeries, robot size is crucial due to the limited workspace. Thus, a robot may be designed under-actuated. Suturing, in contrast, is a complicated task and requires full actuation. This study aims to overcome this shortcoming by implementing an optimization-based algorithm for autonomous suturing for an under-actuated robot. The proposed algorithm approximates the ideal suturing trajectory by slightly reorienting the needle while deviating as little as possible from the ideal, full degree-of-freedom suturing case. The deviation of the path taken by a custom robot with respect to the ideal trajectory varies depending on the suturing starting location within the workspace as well as the needle size. A quantitative analysis reveals that in 13% of the investigated workspace, the accumulative deviation was less than 10 mm. In the remaining workspace, the accumulative deviation was less than 30 mm. Likewise, the accumulative deviation of a needle with a radius of 10 mm was 2.2 mm as opposed to 8 mm when the radius was 20 mm. The optimization-based algorithm maximized the accuracy of a four-DOF robot to perform a path-constrained trajectory and illustrates the accuracy-workspace correlation.

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Early identification and treatment of high-risk plaques before they rupture, and precipitate adverse events constitute a major challenge in cardiology today. Computational simulations are a time- and cost-effective way to study the performance, and to optimize a system. The main objective of this work is to optimize the flow of a novel atraumatic local drug delivery catheter for the treatment of coronary atherosclerosis. The mixing and spreading effectiveness of a drug fluid was analyzed utilizing computational fluid dynamics (CFD) in a coronary artery model. The optimum infusion flow of the nanoparticle-carrying drug fluid was found by maximizing the drug volume fraction and minimizing drug velocity at the artery wall, while maintaining acceptable wall shear stress (WSS). Drug velocities between 15 m/s and 20 m/s are optimum for local drug delivery. The resulting parameters from this study will be used to fabricate customized prototypes for future in-vivo experiments.

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Acute respiratory distress syndrome (ARDS) is a serious respiratory condition that occurs in approximately 10% of patients entering intensive care units around the world, affecting nearly 190,000 patients annually in the United States. Owing to the severity of the condition, conventional methods of oxygenation are often insufficient. However, current alternate methods of oxygenation are associated with contraindications and a mortality rate near 50%. Therefore, a need exists for a safer and more effective method of oxygenation for patients with ARDS. In this work, the feasibility of using intraperitoneal perfusions of oxygen microbubbles-peritoneal microbubble oxygenation (PMO)-to treat lipopolysaccharide-induced ARDS was explored with the objective of showing restoration of normoxic conditions after a single bolus infusion of oxygen microbubbles. Male Wistar rats induced with ARDS via lipopolysaccharide inhalation were treated with PMO at 12-h intervals over a period of 48 h. Their physiological responses were monitored throughout the study, after which necropsy was performed. Response data were then compared with saline control and untreated groups. We conclude that rats experiencing moderate to severe ARDS that were treated with PMO experienced a survival rate 37% higher than animals not given treatment and exhibited increased peripheral blood oxygen saturation when compared with untreated and saline-treated groups. Moreover, those treated with PMO experienced a lower lung wet/dry ratio and less severe lung pathology, indicating a surprising improvement in lung health. Overall, this study demonstrates the ability of PMO to deliver life-sustaining supplemental oxygen to rats suffering from ARDS and warrants further work toward clinical translation.

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INTRODUCTION: Access to high-quality emergency care in low- and middle-income countries (LMIC) is lacking. Many countries utilise a strategy known as "task-shifting" where skills and responsibilities are distributed in novel ways among healthcare personnel. Point-of-care ultrasound (POCUS) has the potential to significantly improve emergency care in LMICs. METHODS: POCUS was incorporated into a training program for a ten-person cohort of non-physician Emergency Care Providers (ECPs) in rural Uganda. We performed a prospective observational evaluation on the impact of a remote, rapid review of POCUS studies on the primary objective of ECP ultrasound quality and secondary objective of ultrasound utilisation. The study was divided into four phases over 11 months: an initial in-person training month, two middle month blocks where ECPs performed ultrasounds independently without remote electronic feedback, and the final months when ECPs performed ultrasounds independently with remote electronic feedback. Quality was assessed on a previously published eight-point ordinal scale by a U.S.-based expert sonographer and rapid standardised feedback was given to ECPs by local staff. Sensitivity and specificity of ultrasound exam findings for the Focused Assessment with Sonography for Trauma (FAST) was calculated. RESULTS: Over the study duration, 1153 ultrasound studies were reviewed. Average imaging frequency per ECP dropped 61% after the initial in-person training month (p = 0.01) when ECPs performed ultrasound independently, but rebounded once electronic feedback was initiated (p = 0.001), with an improvement in quality from 3.82 (95% CI, 3.32-4.32) to 4.68 (95% CI, 4.35-5.01) on an eight-point scale. The sensitivity and specificity of FAST exam during the initial training period was 77.8 (95% CI, 59.2-83.0) and 98.5 (95% CI, 93.3-99.9), respectively. Sensitivity improved 88% compared to independent, non-feedback months whereas specificity was unchanged. CONCLUSIONS: Remotely delivered quality assurance feedback is an effective educational tool to enhance provider skill and foster continued and sustainable use of ultrasound in LMICs.

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Despite the crucial role of RAF kinases in cell signaling and disease, we still lack a complete understanding of their regulation. Heterodimerization of RAF kinases as well as dephosphorylation of a conserved "S259" inhibitory site are important steps for RAF activation but the precise mechanisms and dynamics remain unclear. A ternary complex comprised of SHOC2, MRAS, and PP1 (SHOC2 complex) functions as a RAF S259 holophosphatase and gain-of-function mutations in SHOC2, MRAS, and PP1 that promote complex formation are found in Noonan syndrome. Here we show that SHOC2 complex-mediated S259 RAF dephosphorylation is critically required for growth factor-induced RAF heterodimerization as well as for MEK dissociation from BRAF. We also uncover SHOC2-independent mechanisms of RAF and ERK pathway activation that rely on N-region phosphorylation of CRAF. In DLD-1 cells stimulated with EGF, SHOC2 function is essential for a rapid transient phase of ERK activation, but is not required for a slow, sustained phase that is instead driven by palmitoylated H/N-RAS proteins and CRAF. Whereas redundant SHOC2-dependent and -independent mechanisms of RAF and ERK activation make SHOC2 dispensable for proliferation in 2D, KRAS mutant cells preferentially rely on SHOC2 for ERK signaling under anchorage-independent conditions. Our study highlights a context-dependent contribution of SHOC2 to ERK pathway dynamics that is preferentially engaged by KRAS oncogenic signaling and provides a biochemical framework for selective ERK pathway inhibition by targeting the SHOC2 holophosphatase.

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Purpose/Aim: In developing a novel peritoneal oxygenation therapy, catheters implanted into the peritoneal cavity became obstructed with omental tissue and prevented the infusion and removal of fluid from the peritoneal cavity. The obstruction of peritoneal catheters is a significant failure in researching various peritoneal treatments as further fluid administration is no longer possible. The purpose of this preliminary study was to determine the most effective catheter design for infusion and removal of fluid into the peritoneal cavity of rats. Materials and Methods: Four types of catheters were tested including the Jackson-Pratt, round fluted drain, flat fluted drain, and an original design. Three of each catheter type were surgically placed into the peritoneal cavity of rats (n = 12). In order to test the efficacy of each catheter, saline was infused and extracted twice daily. Catheters were scored on a weighted scale based on the amount of time they remained patent, the subjective force needed for extraction/infusion, and the amount of saline removed. Results: The round and flat fluted drain catheters remained patent for the full duration of the study (12 days) compared to the other models which failed after 7 days. These catheters also yielded a high average for extracted saline volume and an easy extraction/infusion. Conclusions: The round and flat fluted drain catheters were recognized as viable options to be used in rats for peritoneal drain studies of up to 12 days.

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Introduction : Access to high-quality emergency care in low- and middle-income countries (LMIC) is lacking. Many countries utilise a strategy known as "task-shifting" where skills and responsibilities are distributed in novel ways among healthcare personnel. Point-of-care ultrasound (POCUS) has the potential to significantly improve emergency care in LMICs.Methods:POCUS was incorporated into a training program for a ten-person cohort of non-physician Emergency Care Providers (ECPs) in rural Uganda. We performed a prospective observational evaluation on the impact of a remote, rapid review of POCUS studies on the primary objective of ECP ultrasound quality and secondary objective of ultrasound utilisation. The study was divided into four phases over 11 months: an initial in-person training month, two middle month blocks where ECPs performed ultrasounds independently without remote electronic feedback, and the final months when ECPs performed ultrasounds independently with remote electronic feedback. Quality was assessed on a previously published eight-point ordinal scale by a U.S.-based expert sonographer and rapid standardised feedback was given to ECPs by local staff. Sensitivity and specificity of ultrasound exam findings for the Focused Assessment with Sonography for Trauma (FAST) was calculated.Results:Over the study duration, 1153 ultrasound studies were reviewed. Average imaging frequency per ECP dropped 61% after the initial in-person training month (p = 0.01) when ECPs performed ultrasound independently, but rebounded once electronic feedback was initiated (p = 0.001), with an improvement in quality from 3.82 (95% CI, 3.32­4.32) to 4.68 (95% CI, 4.35­5.01) on an eight-point scale. The sensitivity and specificity of FAST exam during the initial training period was 77.8 (95% CI, 59.2­83.0) and 98.5 (95% CI, 93.3­99.9), respectively. Sensitivity improved 88% compared to independent, non-feedback months whereas specificity was unchanged.Conclusions : Remotely delivered quality assurance feedback is an effective educational tool to enhance provider skill and foster continued and sustainable use of ultrasound in LMICs

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Dephosphorylation of the inhibitory "S259" site on RAF kinases (S259 on CRAF, S365 on BRAF) plays a key role in RAF activation. The MRAS GTPase, a close relative of RAS oncoproteins, interacts with SHOC2 and protein phosphatase 1 (PP1) to form a heterotrimeric holoenzyme that dephosphorylates this S259 RAF site. MRAS and SHOC2 function as PP1 regulatory subunits providing the complex with striking specificity against RAF. MRAS also functions as a targeting subunit as membrane localization is required for efficient RAF dephosphorylation and ERK pathway regulation in cells. SHOC2's predicted structure shows remarkable similarities to the A subunit of PP2A, suggesting a case of convergent structural evolution with the PP2A heterotrimer. We have identified multiple regions in SHOC2 involved in complex formation as well as residues in MRAS switch I and the interswitch region that help account for MRAS's unique effector specificity for SHOC2-PP1. MRAS, SHOC2, and PPP1CB are mutated in Noonan syndrome, and we show that syndromic mutations invariably promote complex formation with each other, but not necessarily with other interactors. Thus, Noonan syndrome in individuals with SHOC2, MRAS, or PPPC1B mutations is likely driven at the biochemical level by enhanced ternary complex formation and highlights the crucial role of this phosphatase holoenzyme in RAF S259 dephosphorylation, ERK pathway dynamics, and normal human development.

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Receptor tyrosine kinases (RTKs) are membrane-based sensors that enable rapid communication between cells and their environment. Evidence is now emerging that interdependent regulatory mechanisms, such as membrane trafficking, ubiquitination, proteolysis and gene expression, have substantial effects on RTK signal transduction and cellular responses. Different RTKs exhibit both basal and ligand-stimulated ubiquitination, linked to trafficking through different intracellular compartments including the secretory pathway, plasma membrane, endosomes and lysosomes. The ubiquitin ligase superfamily comprising the E1, E2 and E3 enzymes are increasingly implicated in this post-translational modification by adding mono- and polyubiquitin tags to RTKs. Conversely, removal of these ubiquitin tags by proteases called de-ubiquitinases (DUBs) enables RTK recycling for another round of ligand sensing and signal transduction. The endocytosis of basal and activated RTKs from the plasma membrane is closely linked to controlled proteolysis after trafficking and delivery to late endosomes and lysosomes. Proteolytic RTK fragments can also have the capacity to move to compartments such as the nucleus and regulate gene expression. Such mechanistic diversity now provides new opportunities for modulating RTK-regulated cellular responses in health and disease states.

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We have proposed a long-term, noninvasive, nonrestrictive method of delivering and implanting a biosensor within the body via a swallowable implantation capsule robot (ICR). The design and preliminary validation of the ICR's primary subsystem-the sensor deployment system-is discussed and evidence is provided for major design choices. The purpose of the sensor deployment system is to adhere a small biosensor to the mucosa of the intestine long-term, and the modality was inspired by tapeworms and other organisms that employ a strategy of mechanical adhesion to soft tissue via the combined use of hooks or needles and suckers. Testing was performed to refine the design of the suction and needle attachment as well as the sensor ejection features of the ICR. An experiment was conducted in which needle sharpness, needle length, and vacuum volume were varied, and no statistically significant difference was observed. Finally, preliminary testing, coupled with prior work within a live porcine model, provided evidence that this is a promising approach for implanting a biosensor within the small intestine.

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A wireless medical capsule for measuring the contact pressure between a mobile capsule and the small intestine lumen was developed. Two pressure sensors were used to measure and differentiate the contact pressure and the small intestine intraluminal pressure. After in vitro tests of the capsule, it was surgically placed and tested in the proximal small intestine of a pig model. The capsule successfully gathered and transmitted the pressure data to a receiver outside the body. The measured pressure signals in the animal test were analyzed in the time and frequency domains, and a mathematic model was presented to describe the different factors influencing the contact pressure. A novel signal processing method was applied to isolate the contraction information from the contact pressure. The result shows that the measured contact pressure was 1.08 ± 0.08 kPa, and the small intestine contraction pressure's amplitude and rate were 0.29 ± 0.046 kPa and 12 min-1. Moreover, the amplitudes and rates of pressure from respiration and heartbeat were also estimated. The successful preliminary evaluation of this capsule implies that it could be used in further systematic investigation of small intestine contact pressure on a mobile capsule-shaped bolus.

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Colonoscopy is a diagnostic procedure to detect pre-cancerous polyps and tumours in the colon, and is performed by inserting a long tube equipped with a camera and biopsy tools. Despite the medical benefits, patients undergoing this procedure often complain about the associated pain and discomfort. This discomfort is mostly due to the rough handling of the tube and the creation of loops during the insertion. The overall goal of this work is to minimise the invasiveness of traditional colonoscopy. In pursuit of this goal, this work presents the development of a semi-autonomous colonoscopic robot with minimally invasive locomotion. The proposed robotic approach allows physicians to concentrate mainly on the diagnosis rather than the mechanics of the procedure. In this paper, an innovative locomotion approach for robotic colonoscopy is addressed. Our locomotion approach takes advantage of longitudinal expansion of a latex tube to propel the robot's tip along the colon. This soft and compliant propulsion mechanism, in contrast to minimally invasive mechanisms used in, for example, inchworm-like robots, has shown promising potential. In the preliminary ex vivo experiments, the robot successfully advanced 1.5 metres inside an excised curvilinear porcine colon with average speed of 28 mm/s, and was capable of traversing bends up to 150 degrees. The robot creates less than 6 N of normal force at its tip when it is pressurised with 90 kPa. This maximum force generates pressure of 44.17 mmHg at the tip, which is significantly lower than safe intraluminal human colonic pressure of 80 mmHg. The robot design inherently prevents loop formation in the colon, which is recognised as the main cause of post procedural pain in patients. Overall, the robot has shown great promise in an ex vivo experimental setup. The design of an autonomous control system and in vivo experiments are left as future work.

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