The backbone NMR resonance assignments of the stabilized E. coli ß clamp.

The 81 kDa E. coli ß clamp is a ring-shaped head-to-tail homodimer that encircles DNA and plays a central role in bacterial DNA replication by serving as a processivity factor for DNA polymerases and a binding platform for other DNA replication and repair proteins. Here we report the backbone 1H, 15N, and 13C NMR resonance assignments of the stabilized T45R/S107R ß clamp variant obtained using standard TROSY-based triple-resonance experiments (BMRB 52548). The backbone assignments were aided by 13C and 15N edited NOESY experiments, allowing us to utilize our previously reported assignments of the ß clamp ILV side-chain methyl groups (BMRB 51430, 51431). The backbone assignments of the T45R/S107R ß clamp variant were transferred to the wild-type ß clamp using a minimal set of TROSY-based 15N edited NOESY, NHCO and NHCA experiments (BMRB 52549). The reported backbone and previous ILV side-chain resonance assignments will enable NMR studies of the ß clamp interactions and dynamics using amide and methyl groups as probes.

Public health use and lessons learned from a statewide SARS-CoV-2 wastewater monitoring program (MiNET).

The global SARS-CoV-2 monitoring effort has been extensive, resulting in many states and countries establishing wastewater-based epidemiology programs to address the spread of the virus during the pandemic. Challenges for programs include concurrently optimizing methods, training new laboratories, and implementing successful surveillance programs that can rapidly translate results for public health, and policy making. Surveillance in Michigan early in the pandemic in 2020 highlights the importance of quality-controlled data and explores correlations with wastewater and clinical case data aggregated at the state level. The lessons learned and potential measures to improve public utilization of results are discussed. The Michigan Network for Environmental Health and Technology (MiNET) established a network of laboratories that partnered with local health departments, universities, wastewater treatment plants (WWTPs) and other stakeholders to monitor SARS-CoV-2 in wastewater at 214 sites in Michigan. MiNET consisted of nineteen laboratories, twenty-nine local health departments, 6 Native American tribes, and 60 WWTPs monitoring sites representing 45 % of Michigan's population from April 6 and December 29, 2020. Three result datasets were created based on quality control criteria. Wastewater results that met all quality assurance criteria (Dataset Mp) produced strongest correlations with reported clinical cases at 16 days lag (rho = 0.866, p < 0.05). The project demonstrated the ability to successfully track SARS-CoV-2 on a large, state-wide scale, particularly data that met the outlined quality criteria and provided an early warning of increasing COVID-19 cases. MiNET is currently poised to leverage its competency to complement public health surveillance networks through environmental monitoring for new and emerging pathogens of concern and provides a valuable resource to state and federal agencies to support future responses.

Revisiting the Roles of Catalytic Residues in Human Ornithine Transcarbamylase.

Human ornithine transcarbamylase (hOTC) is a mitochondrial transferase protein involved in the urea cycle and is crucial for the conversion of toxic ammonia to urea. Structural analysis coupled with kinetic studies of Escherichia coli, rat, bovine, and other transferase proteins has identified residues that play key roles in substrate recognition and conformational changes but has not provided direct evidence for all of the active residues involved in OTC function. Here, computational methods were used to predict the likely active residues of hOTC; the function of these residues was then probed with site-directed mutagenesis and biochemical characterization. This process identified previously reported active residues, as well as distal residues that contribute to activity. Mutation of active site residue D263 resulted in a substantial loss of activity without a decrease in protein stability, suggesting a key catalytic role for this residue. Mutation of predicted second-layer residues H302, K307, and E310 resulted in significant decreases in enzymatic activity relative to that of wild-type (WT) hOTC with respect to l-ornithine. The mutation of fourth-layer residue H107 to produce the hOTC H107N variant resulted in a 66-fold decrease in catalytic efficiency relative to that of WT hOTC with respect to carbamoyl phosphate and a substantial loss of thermal stability. Further investigation identified H107 and to a lesser extent E98Q as key residues involved in maintaining the hOTC quaternary structure. This work biochemically demonstrates the importance of D263 in hOTC catalytic activity and shows that residues remote from the active site also play key roles in activity.

Fit-for-Purpose Ki-67 Immunohistochemistry Assays for Breast Cancer.

New therapies are being developed for breast cancer, and in this process, some "old" biomarkers are reutilized and given a new purpose. It is not always recognized that by changing a biomarker's intended use, a new biomarker assay is created. The Ki-67 biomarker is typically assessed by immunohistochemistry (IHC) to provide a proliferative index in breast cancer. Canadian laboratories assessed the analytical performance and diagnostic accuracy of their Ki-67 IHC laboratory-developed tests (LDTs) of relevance for the LDTs' clinical utility. Canadian clinical IHC laboratories enrolled in the Canadian Biomarker Quality Assurance Pilot Run for Ki-67 in breast cancer by invitation. The Dako Ki-67 IHC pharmDx assay was employed as a study reference assay. The Dako central laboratory was the reference laboratory. Participants received unstained slides of breast cancer tissue microarrays with 32 cases and performed their in-house Ki-67 assays. The results were assessed using QuPath, an open-source software application for bioimage analysis. Positive percent agreement (PPA, sensitivity) and negative percent agreement (NPA, specificity) were calculated against the Dako Ki-67 IHC pharmDx assay for 5%, 10%, 20%, and 30% cutoffs. Overall, PPA and NPA varied depending on the selected cutoff; participants were more successful with 5% and 10%, than with 20% and 30% cutoffs. Only 4 of 16 laboratories had robust IHC protocols with acceptable PPA for all cutoffs. The lowest PPA for the 5% cutoff was 85%, for 10% was 63%, for 20% was 14%, and for 30% was 13%. The lowest NPA for the 5% cutoff was 50%, for 10% was 33%, for 20% was 50%, and for 30% was 57%. Despite many years of international efforts to standardize IHC testing for Ki-67 in breast cancer, our results indicate that Canadian clinical LDTs have a wide analytical sensitivity range and poor agreement for 20% and 30% cutoffs. The poor agreement was not due to the readout but rather due to IHC protocol conditions. International Ki-67 in Breast Cancer Working Group (IKWG) recommendations related to Ki-67 IHC standardization cannot take full effect without reliable fit-for-purpose reference materials that are required for the initial assay calibration, assay performance monitoring, and proficiency testing.

Towards understanding and improving medication safety for patients with mental illness in primary care: A multimethod study.

INTRODUCTION: Medication safety incidents have been identified as an important target to improve patient safety in mental healthcare. Despite this, the causes of preventable medication safety incidents affecting patients with mental illness have historically been poorly understood, with research now addressing this knowledge gap through a healthcare professional lens. However, patients and carers can also provide complimentary insight into safety issues, and as key stakeholders in healthcare, it is vital to consider their needs when designing effective interventions. METHODS: A two-stage approach was adopted by (i) conducting three focus groups (FG) comprising 13 patients with mental illness and their carers to develop a holistic picture of medication safety in primary care with extraction of themes guided by the P-MEDS framework; (ii) conducting two separate nominal group consensus workshops with seven patients with mental illness/carers and seven healthcare professionals to identify priority areas for targeted interventions. RESULTS: Seven themes were identified in the FGs: communication; trust, involvement and respect; continuity and support; access; the healthcare professional; the patient and carer; and the organisation. Priority areas identified for intervention by key stakeholders included improving communication within and between clinical services, enhancing patient support with holistic continuity of care, maximising shared decision-making and empowerment, ensuring timely access to medicines and services, strengthening healthcare professional knowledge regarding mental illnesses and associated medications, and increasing patient dignity and respect. CONCLUSION: This study has developed a holistic picture of contributors to medication safety incidents affecting patients with mental illness in primary care. This theory was then used by key stakeholders to inform and generate priority recommendations for targeted interventions. These findings can be used to inform future intervention research, as they consider the needs of those who access or work within primary care services. PATIENT OR PUBLIC CONTRIBUTION: An advisory group consisting of three expert patients with lived experience of mental illness was consulted on the design of both stages of this study. Patients with mental illness and/or their carers were recruited and participated in both stages of this study. Patients/carers aided with data analysis and interpretation during the patient/carer nominal group consensus workshop.

Cooled feline intestine and fresh intestine did not differ in enterotomy leak pressure testing or in gross wall thickness measurement.

OBJECTIVE: To report gross anatomical gastrointestinal measurements and compare enterotomy leak pressures between fresh and cooled feline cadavers. STUDY DESIGN: Ex vivo, randomized study. ANIMALS: Fresh feline cadavers (n = 20). METHODS: Jejunal segments (8 cm) were harvested on the same day as euthanasia. From each cadaver, one segment was randomly assigned to control (C), fresh enterotomy (FE), and cooled enterotomy (CE) groups. Enterotomy construction and leak testing were performed within 12 h of euthanasia for the C and FE groups and after 17-29 h of cooling for the CE group. Initial leak pressure (ILP) and maximum intraluminal pressure (MIP) were compared. Gastrointestinal wall thickness and intraluminal diameter were measured on harvested applicable gastrointestinal divisions at up to three time points: day 1 fresh, day 2 cooled, and day 3 cooled. RESULTS: The mean (± SD) ILPs for the C, FE, and CE constructs were 600 (± 0.0), 200.3 (± 114.7), and 131.3 (± 92.6) mmHg, respectively. The C ILP was higher (p < .001) than the FE and CE ILP. The ILP (p = .11) and the MIP (p = .21) did not differ between the FE and CE constructs. Wall thickness (measured in mm) did not differ between duodenum day 1 fresh and day 2 cooled groups (p = .18) or between any jejunum day groups (p = .86). The intraluminal diameters (mean ± SD) for the duodenum, jejunum, and ileum were 5.7 (± 0.7), 5.8 (± 0.8), and 7.2 (± 2.2) mm, respectively. CONCLUSION: No difference was appreciated between FE and CE ILP and MIP. Wall thickness measurements did not differ between days for duodenum or jejunum. CLINICAL RELEVANCE: Cadaveric feline intestine cooled for up to 29 h may be used for determining intestinal leak pressures.

Clinical manifestations of chronic pancreatitis in English cocker spaniels.

BACKGROUND: Chronic pancreatitis (CP) is common in English cocker spaniels (ECS). It is histologically similar to IgG4-related disease (IgG4-RD) in humans and is characterized by duct destruction, interlobular fibrosis, and dense periductular and perivenous lymphocytic aggregates. However, the clinical manifestations of CP in ECS have not been previously described. OBJECTIVES: Characterize the clinical manifestations of CP in a group of ECS, including similarities and differences to IgG4-RD in humans. ANIMALS: One-hundred four ECS with CP and 44 client owned control ECS without CP (both healthy and diseased controls). METHODS: Affected dogs were divided into 2 groups according to the methods used to diagnose CP. Case records were searched for signalment, clinical, and clinicopathological findings, and evidence of keratoconjunctivitis sicca (KCS), proteinuria, other immune-mediated diseases, and anal sacculitis. RESULTS: Involvement of other organs was common. Affected ECS presented with a high frequency of KCS (n = 49), proteinuria (n = 47), anal gland disease (n = 36), atopy (n = 21), and other immune-mediated diseases (n = 16). Those with parti-color hair coats, particularly blue roan, had a strong association with CP, suggesting a link between coat color and autoimmune conditions in this breed. CONCLUSIONS AND CLINICAL IMPORTANCE: English cocker spaniels with CP show clinical similarities to humans with IgG4-RD and common involvement of other organs. Clinicians should evaluate affected Cocker Spaniels for proteinuria, keratoconjunctivitis sicca, and other potential immune-mediated diseases.

A numerical study of dehydration induced fracture toughness degradation in human cortical bone.

A 2D plane strain extended finite element method (XFEM) model was developed to simulate three-point bending fracture toughness tests for human bone conducted in hydrated and dehydrated conditions. Bone microstructures and crack paths observed by micro-CT imaging were simulated using an XFEM damage model. Critical damage strains for the osteons, matrix, and cement lines were deduced for both hydrated and dehydrated conditions and it was found that dehydration decreases the critical damage strains by about 50%. Subsequent parametric studies using the various microstructural models were performed to understand the impact of individual critical damage strain variations on the fracture behavior. The study revealed the significant impact of the cement line critical damage strains on the crack paths and fracture toughness during the early stages of crack growth. Furthermore, a significant sensitivity of crack growth resistance and crack paths on critical strain values of the cement lines was found to exist for the hydrated environments where a small change in critical strain values of the cement lines can alter the crack path to give a significant reduction in fracture resistance. In contrast, in the dehydrated state where toughness is low, the sensitivity to changes in critical strain values of the cement lines is low. Overall, our XFEM model was able to provide new insights into how dehydration affects the micromechanisms of fracture in bone and this approach could be further extended to study the effects of aging, disease, and medical therapies on bone fracture.

Performance time and leak pressure of hand-sewn and skin staple intestinal anastomoses and enterotomies in cadaveric cats.

OBJECTIVE: To compare time to construct completion and leak testing between hand-sewn and skin staple anastomoses and enterotomies in cats. STUDY DESIGN: Ex vivo, randomized study. ANIMALS: Fresh feline cadavers (n = 20). METHODS: Jejunal segments (8 cm) were harvested and tested on the same day as euthanasia. From each cadaver, one segment was randomly assigned to control (C), hand-sewn enterotomy (HSE), and skin staple enterotomy (SSE) groups, and two segments were randomly assigned to hand-sewn anastomosis (HSA) and skin staple anastomosis (SSA) groups. Construct completion time, initial leak pressure (ILP), and maximum intraluminal pressure were compared. Leakage location was reported. RESULTS: Mean time (s) ± SD was longer (p < .001) for HSA (317.0 ± 50.9) than SSA (160.8 ± 13.1) and for HSE (172.0 ± 36.5) than SSE (20.3 ± 5.0). ILP (mean ± SD) for C (600.0 mmHg ±0.0) was higher (p < .001) than all constructs. ILP (mean ± SD) for SSA (124.2 mmHg ±83.7) was not different (p = .49) than HSA (86.1 ± 51.9), but HSE (200.3 ± 114.7) was higher (p < .001) than SSE (32.2 ± 39.7). Immediate leakage from the center of enterotomy closure was observed in 7/20 SSE. CONCLUSIONS: HSA construct completion took twice as long as SSA with no difference in intraluminal pressures. Although HSE construct completion took 8x as long as SSE, HSE had higher intraluminal pressures. CLINICAL SIGNIFICANCE: In cats, SSA may be an alternative to HSA for intestinal anastomosis, but SSE is not recommended as an alternative to HSE for intestinal enterotomy closure.

Impact of heavy alcohol consumption on cortical bone mechanical properties in male rhesus macaques.

Chronic heavy alcohol consumption may influence the skeleton by suppressing intracortical bone remodeling which may impact the quality of bone and its mechanical properties. However, this aspect has not been thoroughly assessed in either humans or animal models whose cortical bone microstructure resembles the microstructure of human cortical bone. The current study is the first to investigate the effects of chronic heavy alcohol consumption on various mechanical properties of bone in a non-human primate model with intracortical remodeling. Male rhesus macaques (5.3 years old at the initiation of treatment) were induced to drink alcohol and then given the choice to voluntarily self-administer water or ethanol (4 % w/v) for approximately 14 months, followed by three abstinence phases (lasting 34, 41, and 39-46 days) with approximately 3 months of ethanol access in between. During the initial 14 months of open-access, monkeys in the alcohol group consumed an average of 2.9 ± 0.8 g/kg/d ethanol (mean ± SD) resulting in a blood ethanol concentration of 89 ± 47 mg/dl in longitudinal samples taken at 7 h after the daily sessions began. To understand the impact of alcohol consumption on material properties, various mechanical tests were conducted on the distal tibia diaphysis of 2-5 monkeys per test group, including dynamic mechanical analysis (DMA) testing, nano-indentation, microhardness testing, compression testing, and fracture resistance curve (R-curve) testing. Additionally, compositional analyses were performed using Fourier-transform infrared (FTIR) spectroscopy. Significant differences in microhardness, compressive stress-strain response, and composition were not observed with alcohol consumption, and only minor differences were detected in hardness and elastic modulus of the matrix and osteons from nanoindentation. Furthermore, the R-curves of both groups overlapped, with similar crack initiation toughness, despite a significant decrease in crack growth toughness (p = 0.032) with alcohol consumption. However, storage modulus (p = 0.029) and loss factor (p = 0.015) from DMA testing were significantly increased in the alcohol group compared to the control group, while loss modulus remained unchanged. These results indicate that heavy alcohol consumption may have only a minor influence on the material properties and the composition of cortical bone in young adult male rhesus macaques.

CE: Our Journey to Pass a Surgical Plume Evacuation Law.

ABSTRACT: Surgical plume is produced when heat-generating instruments are used to cut, cauterize, or vaporize tissue during surgery. Surgical plume can rapidly diffuse, and without the use of plume evacuators to ensure it is captured, filtered, and removed, health care workers and patients are at risk for inhalation exposure to airborne contaminants, including smoke and biological hazards. The use of surgical plume evacuators in health care facilities is inconsistent across the United States. This article describes the experience of two nurses who led a grassroots coalition to pass a surgical plume evacuation law in Illinois. It also outlines strategies they used in the legislative process that nurses can apply to their own efforts to advocate for health policy change at the state level.

Effect of gamma irradiation and supercritical carbon dioxide sterilization with Novakill™ or ethanol on the fracture toughness of cortical bone.

Sterilization of structural bone allografts is a critical process prior to their clinical use in large cortical bone defects. Gamma irradiation protocols are known to affect tissue integrity in a dose dependent manner. Alternative sterilization treatments, such as supercritical carbon dioxide (SCCO2 ), are gaining popularity due to advantages such as minimal exposure to denaturants, the lack of toxic residues, superior tissue penetration, and minor impacts on mechanical properties including strength and stiffness. The impact of SCCO2 on the fracture toughness of bone tissue, however, remains unknown. Here, we evaluate crack initiation and growth toughness after 2, 6, and 24 h SCCO2 -treatment using Novakill™ and ethanol as additives on ~11 samples per group obtained from a pair of femur diaphyses of a canine. All mechanical testing was performed at ambient air after 24 h soaking in Hanks' balanced salt solution (HBSS). Results show no statistically significant difference in the failure characteristics of the Novakill™-treated groups whereas crack growth toughness after 6 and 24 h of treatment with ethanol significantly increases by 37% (p = .010) and 34% (p = .038), respectively, compared to an untreated control group. In contrast, standard 25 kGy gamma irradiation causes significantly reduced crack growth resistance by 40% (p = .007) compared to untreated bone. FTIR vibrational spectroscopy, conducted after testing, reveals a consistent trend of statistically significant differences (p < .001) with fracture toughness. These trends align with variations in the ratios of enzymatic mature to immature crosslinks in the collagen structure, suggesting a potential association with fracture toughness. Additional Raman spectroscopy after testing shows a similar trend with statistically significant differences (p < .005), which further supports that collagen structural changes occur in the SCF-treated groups with ethanol after 6 and 24 h. Our work reveals the benefits of SCCO2 sterilization compared to gamma irradiation.

Gastrointestinal thickness, duration, and leak pressure of five intestinal anastomosis techniques in cats.

OBJECTIVE: To compare time to construct completion and resistance to leakage for five intestinal anastomosis techniques in cats and to report normal feline gastrointestinal thickness. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Grossly normal intestinal segments (n = 120) from 10 fresh cat cadavers. METHODS: A total of 8 cm segments of fresh feline cadaveric intestine were collected, and mural thickness was recorded. Segments were randomly allocated between a control group (n = 20 segments) and five treatment groups (20 segments/group with 2 segments/construct = 10 constructs per group): (1) hand-sewn anastomosis - simple interrupted (HSA-SI), (2) hand-sewn anastomosis - simple continuous (HSA-SC), (3) functional end-to-end stapled anastomosis (FEESA), (4) functional end-to-end stapled anastomosis with oversew (FEESA-O), (5) skin stapled anastomosis (SS). Time to construct completion, leakage location, initial leak pressure (ILP), and maximum intraluminal pressure (MIP) were compared. RESULTS: Mean mural thickness ± SD (mm) for the stomach, duodenum, jejunum, and ileum were 1.66 ± 0.28, 2.05 ± 0.18, 2.28 ± 0.30, and 2.11 ± 0.39, respectively. ILPs (mean ± SD) for HSA-SI (165 ± 122 mmHg), HSA-SC (149 ± 83), FEESA-O (63 ± 25, FEESA (84 ± 59), SS (77 ± 56), and control segments (>500) were compared. There was no statistically significant difference in ILP (p > .08) or MIP (p > .084) between any treatment groups. Nonoversewn FEESAs were 2.4 times faster to perform compared to oversewn FEESA and SS groups, and 4.7 times faster than HSA (p < .001). CONCLUSION: All anastomosis techniques provide resistance to leakage that is supraphysiological to that of the normal maximum intraluminal pressure. HSA take longer to complete than stapled anastomoses. CLINICAL SIGNIFICANCE: All anastomotic techniques may be appropriate in cats. Hand-sewn anastomoses result in a longer surgical time.

Exploring the challenges faced by foundation doctors when prescribing high risk medicines safely during the on-call period: A qualitative study.

AIMS: Errors with prescribing high-risk medicines (HRMs) have a greater propensity to cause harm than with non-HRMs. Prescribing errors arise due to multiple factors and it can be particularly challenging for junior doctors to prescribe safely during the on-call period. Knowledge regarding the challenges of prescribing HRM during the on-call period would be useful to target preventative interventions. The aim of this study was to explore the challenges encountered by foundation doctors (doctors who have graduated medical school within the last 2 years) when prescribing specific HRMs (anticoagulants, insulin and opioids) safely during the on-call period. METHODS: Six focus groups exploring the challenges of prescribing HRMs safely during the on-call period were conducted, 3 with foundation year 1 and 3 with foundation year 2 doctors from across 3 different hospitals. A thematic framework analysis based on the London Protocol was conducted. RESULTS: Doctors described multiple challenges to prescribing HRMs safely during the on-call period including a lack of prescribing support, nursing pressure, complex prescribing tasks, unknown patients as well as individual factors such as lack of knowledge and tiredness. Many of these factors exist to some extent during the day, yet the nature of the on-call period as a fast-paced environment heightened the challenges that prescribers faced. CONCLUSION: There are multiple challenges experienced by foundation doctors when prescribing HRMs during the on-call period. The potentially devastating consequences of errors with HRMs means that closer attention and more concern from healthcare professionals, researchers and policymakers is required to improve safe prescribing of HRMs in hospitals.

Complications and outcome following staphylectomy and folded flap palatoplasty in dogs with brachycephalic obstructive airway syndrome.

OBJECTIVE: To compare the prevalence of pre-, intra-, and postoperative variables and complications associated with staphylectomy (S) and folded flap palatoplasty (FFP). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Client-owned dogs (n = 124). METHODS: Medical records of S and FFP dogs from a veterinary teaching hospital were reviewed between July 2012 and December 2019. Signalment, clinical pre-, intra-, and postoperative data were collected and reviewed. Median (interquartile range) was reported. RESULTS: A total of 124 dogs among 14 breeds underwent surgical treatment for an elongated soft palate with either a S (n = 64) or FFP (60). FFP dogs without concurrent non-airway procedures were associated with longer duration of surgery (p = .02; n = 63; S, median = 51 min [34-85]; FFP, median = 75 min [56.25-94.5]) and anesthesia (p = .02; n = 63; S, median = 80 min [66-125]; FFP, median = 111 min [91-140.8]). Neither soft palate surgery was associated with the occurrence of anesthetic complications (p = .30; 99/120; S, 49; FFP, 50), postoperative regurgitation (p = .18; 27/124; S, 17; FFP, 10), or with hospitalization duration (p = .94; n = 124; S, median = 1 day [1]; FFP, median = 1 [1]). Postoperative aspiration pneumonia (9/124; S, 4; FFP, 5) and major complications were rare (5/124; S, 3; FFP, 2). CONCLUSION: S and FFP had similar anesthetic and perioperative complications, although FFP dogs had longer anesthetic and operative times. CLINICAL SIGNIFICANCE: Although FFP took longer, no other clinically significant differences were appreciated between S and FFP procedures. Because of limitations inherent in study design, surgeons should continue to use clinical judgment when deciding on a procedure.

Implementing flexible endoscopic evaluation of swallow screening within annual cancer surveillance appointments to monitor for late-stage radiation-induced dysphagia: A feasibility study.

BACKGROUND: Late-stage progressive decline of swallowing function after radiotherapy for head and neck cancer (HNC) is often difficult to monitor. This study examined the feasibility and clinical outcomes of speech-language pathology implementing flexible endoscopic evaluation of swallow (FEES) screening during annual cancer surveillance visits to monitor late-stage swallowing function. METHODS: Patients >2 years post treatment who attended routine oncological visits underwent FEES screening. Feasibility (service data, stakeholder survey) and swallowing outcomes (oral intake, secretions, internal lymphedema, penetration-aspiration, and residue) were collected. RESULTS: Screening was completed with 70% (50/71) of eligible patients. Medical staff and speech-language pathologists indicated the protocol was worthwhile and achievable to incorporate into practice. Almost all patients were willing to complete the protocol annually. FEES outcomes identified 84% with dysphagia versus only 26% self-reported dysphagia. CONCLUSION: Findings indicate FEES screening incorporated into annual oncological reviews is feasible and effective at monitoring late-stage swallowing function following HNC.

Use of real-time near-infrared fluorescence to assess gastric viability in dogs with gastric dilatation volvulus: A case-control study.

OBJECTIVE: To describe near-infrared fluorescence (NIRF) for assessment of gastric viability and describe NIRF's influence on the surgeon's operative strategy in dogs with gastric dilatation and volvulus (GDV). STUDY DESIGN: Prospective clinical trial. ANIMALS: Twenty dogs with GDV and 20 systemically healthy dogs. METHODS: Following gastric derotation, the surgeon's subjective assessment of gastric viability was recorded prior to near-infrared imaging. Changes in the surgeon's initial assessment of viability based on the visual pattern of gastric fluorescence was recorded. If nonviable (lack of defined vessels), a partial gastrectomy was performed and submitted for histopathology. The stapled gastrectomy line was imaged. Viable (defined vessels) and nonviable fluorescence intensities were compared with healthy dogs undergoing surgery for nongastrointestinal disease. RESULTS: Subjective assessment diagnosed 17 viable and three nonviable GDVs (2 fundi; 1 cardia). Near-infrared imaging demonstrated nonviable gastric fluorescence in 4 dogs (3 fundi/cardia; 1 fundus). The surgeon's margins for resection were altered in 3/20 dogs. Fluorescence intensity (cardia, fundus, body, pylorus) was lower in GDV viable (30.59%, p = .04; 38.17%, p < .01; 51.18%, p < .01; 44.12%, p= .01) and nonviable (11.00%, p < .01; 4.33%, p < .01; 57.67%, p = .22; 54.33%, p = .72) dogs compared to healthy controls (44.7%, 70.05%, 84.00%, 63.95%). Fundic fluorescence was less in nonviable gastric tissue in comparison with viable gastric tissue (p = .03). Fluorescence of the gastrectomy staple line approximated that of viable tissue. CONCLUSION: Near-infrared fluorescence can identify histologically confirmed nonviable gastric tissue. CLINICAL SIGNIFICANCE: These results provide enough evidence to support the implementation of NIRF as an adjunct to gross examination of the gastric wall in dogs with GDV.

unDerstandIng the cauSes of mediCation errOrs and adVerse drug evEnts for patients with mental illness in community caRe (DISCOVER): a qualitative study.

Background: It is estimated that 237 million medication errors occur in England each year with a significant number occurring in the community. Our understanding of the causes of preventable medication errors and adverse drug events (ADE) affecting patients with mental illness is limited in this setting. Better understanding of the factors that contribute to errors can support the development of theory-driven improvement interventions. Methods: Remote qualitative semi-structured interviews with 26 community-based healthcare professionals in England and Wales were undertaken between June-November 2022. Recruitment was undertaken using purposive sampling via professional networks. Interviews were guided by the critical incident technique and analysed using the framework method. Any data that involved speculation was not included in the analysis. Independent analysis was carried out by the research team to extract themes guided by the London Protocol. Results: A total of 43 medication errors and 12 preventable ADEs were discussed, with two ADEs having an unknown error origin. Prescribing errors were discussed most commonly (n = 24), followed by monitoring errors (n = 8). Six contributory factor themes were identified: the individual (staff); the work environment; the teams/interfaces; the organisation and management; the patient; and the task and technology. The individual (staff) factors were involved in just over 80% of all errors discussed. Participants reported a lack of knowledge regarding psychotropic medication and mental illnesses which accompanied diffusion of responsibility. There were difficulties with team communication, particularly across care interfaces, such as ambiguity/brevity of information being communicated and uncertainty concerning roles which created confusion amongst staff. Unique patient social/behavioural contributory factors were identified such as presenting with challenging behaviour and complex lifestyles, which caused difficulties attending appointments as well as affecting overall clinical management. Conclusion: These findings highlight that the causes of errors are multifactorial with some unique to this patient group. Key areas to target for improvement include the education/training of healthcare professionals regarding neuropharmacology/mental illnesses and enhancing communication across care interfaces. Future research should explore patient perspectives regarding this topic to help develop a holistic picture. These findings can be used to guide future intervention research to ameliorate medication safety challenges for this patient group.

Biochemical Activity of 17 Cancer-Associated Variants of DNA Polymerase Kappa Predicted by Electrostatic Properties.

DNA damage and repair have been widely studied in relation to cancer and therapeutics. Y-family DNA polymerases can bypass DNA lesions, which may result from external or internal DNA damaging agents, including some chemotherapy agents. Overexpression of the Y-family polymerase human pol kappa can result in tumorigenesis and drug resistance in cancer. This report describes the use of computational tools to predict the effects of single nucleotide polymorphism variants on pol kappa activity. Partial Order Optimum Likelihood (POOL), a machine learning method that uses input features from Theoretical Microscopic Titration Curve Shapes (THEMATICS), was used to identify amino acid residues most likely involved in catalytic activity. The µ4 value, a metric obtained from POOL and THEMATICS that serves as a measure of the degree of coupling between one ionizable amino acid and its neighbors, was then used to identify which protein mutations are likely to impact the biochemical activity. Bioinformatic tools SIFT, PolyPhen-2, and FATHMM predicted most of these variants to be deleterious to function. Along with computational and bioinformatic predictions, we characterized the catalytic activity and stability of 17 cancer-associated DNA pol kappa variants. We identified pol kappa variants R48I, H105Y, G147D, G154E, V177L, R298C, E362V, and R470C as having lower activity relative to wild-type pol kappa; the pol kappa variants T102A, H142Y, R175Q, E210K, Y221C, N330D, N338S, K353T, and L383F were identified as being similar in catalytic efficiency to WT pol kappa. We observed that POOL predictions can be used to predict which variants have decreased activity. Predictions from bioinformatic tools like SIFT, PolyPhen-2, and FATHMM are based on sequence comparisons and therefore are complementary to POOL but are less capable of predicting biochemical activity. These bioinformatic and computational tools can be used to identify SNP variants with deleterious effects and altered biochemical activity from a large data set.