Presentation, diagnosis, and outcomes of cats undergoing surgical treatment of ectopic ureters.

OBJECTIVE: To describe the signalment, treatment, complications, and outcomes of cats treated surgically for ectopic ureters. STUDY DESIGN: Retrospective, multi-institutional study. ANIMALS: Twelve client-owned cats. METHODS: Medical records of cats diagnosed with unilateral or bilateral ectopic ureters were reviewed and analyzed. Data reported included signalment, clinical signs, diagnostics, open celiotomy, or cystoscopic surgical interventions, and outcomes. RESULTS: Seven of the 12 cats in the study population were female or female spayed and the median age at time of presentation was 4 years, with an interquartile range (IQR) of 6 months-14 years. Presurgical diagnostic imaging diagnosed ectopic ureters by abdominal ultrasound (8/10), contrast enhanced computed tomography (3/3), fluoroscopic urography (3/4), or cystoscopy (6/7). Eight of 12 cats had extramural ectopic ureters and six cats were affected bilaterally. Eight affected cats underwent ureteroneocystostomy, one cat underwent neoureterostomy, two cats underwent cystoscopic laser ablation, and one cat underwent nephroureterostomy. Immediate postoperative complications occurred in three cats; one cat required additional surgical intervention. Short-term complications occurred in three cats, and long-term complications in two cats. All cats that underwent surgical or cystoscopic intervention had improvement of their urinary incontinence scores, with complete resolution in 11 cats. CONCLUSION: Surgical correction of ectopic ureters in cats is associated with good long-term outcomes. Ectopic ureters in cats are commonly extramural and bilateral. Postoperative outcomes were acceptable and there were few postoperative complications, with varying forms of surgical correction. CLINICAL SIGNIFICANCE: Ectopic ureters in cats are rare but urinary incontinence can be corrected or improved successfully with surgery.

Prospective, randomized, clinical trial on the effects of laparoscopic insufflation pressures on portal pressures in dogs.

OBJECTIVE: The adverse effects of intra-abdominal pressure from capnoperitoneum on cardiovascular and pulmonary systems have been well documented, but the effects on portal pressures in dogs with various insufflation pressures is poorly defined. The aim of the present study was to measure the effect of a range of insufflation pressures on the portal pressure, using direct pressure measurements in patients undergoing laparoscopy. STUDY DESIGN: Clinical randomized prospective study. ANIMALS: Nine client-owned dogs undergoing routine laparoscopy. METHODS: Two rounds of direct portal pressure assessments were performed, at insufflation pressures of 0, 6, 10, and 14 mmHg in a predetermined randomized sequence. The data were analyzed for effects of insufflation pressure, hemodynamic alterations, and round. A best-fit exponential model of the relationship between portal pressure and insufflation pressure was created. RESULTS: Portal pressure increased by 38% at 6 mmHg, 95% at 10 mmHg, and 175% at 14 mmHg compared to baseline. Portal pressure increased at an average rate of 7.45% per mmHg of insufflation pressure. Effects of weight, weight/insufflation pressure interaction, and round of insufflation were not statistically significant. No systemic hemodynamic adverse events were observed. CONCLUSION: Portal pressure increased as insufflation pressure increased. There was no clinically significant difference in baseline portal pressure between rounds of insufflation. CLINICAL SIGNIFICANCE: This exponential model of portal pressure supports the use of the minimum insufflation pressure to allow visualization during laparoscopy. The return of portal pressure to baseline following desufflation supports the comparison of portal pressure measurements before and after laparoscopic shunt attenuation.

Expanding the Tool Kit of Automated Flow Synthesis: Development of In-line Flash Chromatography Purification.

Recent advancements in in-line extraction and purification technology have enabled complex multistep synthesis in continuous flow reactor systems. However, for the large scope of chemical reactions that yield mixtures of products or residual starting materials, off-line purification is still required to isolate the desired compound. We present the in-line integration of a commercial automated flash chromatography system with a flow reactor for the continuous synthesis and isolation of product(s). A proof-of-principle study was performed to validate the system and test the durability of the column cartridges, performing an automated sequence of 100 runs over 2 days. Three diverse reaction systems that highlight the advantages of flow synthesis were successfully applied with in-line normal- or reversed-phase flash chromatography, continuously isolating products with 97-99% purity. Productivity of up to 9.9 mmol/h was achieved, isolating gram quantities of pure product from a feed of crude reaction mixture. Herein, we describe the development and optimization of the systems and suggest guidelines for selecting reactions well suited to in-line flash chromatography.

Development of autotaxin inhibitors: A series of tetrazole cinnamides.

A series of inhibitors of Autotaxin (ATX) have been developed from a high throughput screening hit, 1a, which shows an alternative binding mode to known catalytic site inhibitors. Selectivity over the hERG channel and microsomal clearance were dependent on the lipophilicity of the compounds, and this was optimised by reduction of clogD whilst maintaining high affinity ATX inhibition. Compound 15a shows good oral exposure, and concentration dependent inhibition of formation of LPA in vivo, as shown in pharmacokinetic-pharmacodynamic (PK/PD) experiments.

Outcomes after transperitoneal laparoscopic ureteronephrectomy for the treatment of primary renal neoplasia in seven dogs.

OBJECTIVE: To describe the perioperative characteristics and outcomes in dogs that underwent transperitoneal laparoscopic ureteronephrectomy (TLU) for primary renal neoplasia. STUDY DESIGN: Short case series. ANIMALS: Seven client-owned dogs. METHODS: Medical records were reviewed and data extracted regarding perioperative characteristics and animal outcomes. TLU was performed using a single-port + 1 or multiple port techniques. Hemostatic clips or a vessel-sealing device were used for occlusion of renal hilar vessels. The ureter was occluded and transected close to the ureterovesicular junction and the tumor was placed in a specimen retrieval bag for extraction from the abdomen. RESULTS: Preoperative contrast enhanced computed tomography (CECT) was performed in 6/7 dogs. Median estimated tumor volume measured from abdominal CECT removed by TLU was 32.42 cm3 (interquartile range [IQR] 14.76-94.85). Median surgery time for TLU was 90 minutes (IQR 85-105). In one dog, elective conversion to open laparotomy was performed due to large tumor size. Median time to discharge was 31 hours (IQR 24-48). No major perioperative complications occurred and all dogs survived to discharge. Progression free survival in four dogs was 422 days (IQR 119-784). CONCLUSION: TLU was performed for the extirpation of modest sized primary renal tumors with acceptable perioperative outcomes and a low complication rate. CLINICAL RELEVANCE: TLU may be considered for the treatment of selected cases of primary renal neoplasia in dogs.

Clinical findings and outcomes of 153 dogs surgically treated for intestinal intussusceptions.

OBJECTIVE: To describe perioperative characteristics and outcomes of dogs surgically treated for intestinal intussusception. STUDY DESIGN: Multi-institutional, retrospective study. ANIMALS: One hundred fifty-three client-owned dogs with intestinal intussusception. METHODS: Dogs were included when they had undergone surgical treatment of a confirmed intestinal intussusception. Medical records were reviewed for demographics and clinical data, including surgical complications (graded 1-4). Follow-up was obtained via telephone interview with owners and referring veterinarians. RESULTS: Dogs had a median age of 10 months (range, 2-156), and the most common location for intussusception was ileocolic (66/153 [43%]). Most cases had no identifiable cause (104/155 [67%]). Intestinal resection and anastomosis (IRA) was performed in 129 of 153 (84%) dogs; enteroplication was performed in 28 of 153 (18%) dogs, including 13 with and 15 without IRA. Intraoperative complications occurred in 10 of 153 (7%) dogs, all involving intestinal damage during attempted manual reduction. The median duration of follow-up after discharge was 334 days (interquartile range, 15-990; range, 1-3302). Postoperative complications occurred in 53 of 153 (35%) dogs, including 22 of 153 (14%) with severe (grade 3 or 4) events. Diarrhea, regurgitation, and septic peritonitis were the most common postoperative complications; intussusception recurred in four of 153 (3%) dogs, all within 72 hours postoperatively. Fourteen-day postoperative mortality rate was 6%. CONCLUSION: Surgical treatment of intestinal intussusception was curative in most dogs, even when an underlying cause was not identified. Surgical complications were common, including a 14% risk of life-threatening short-term complications. CLINICAL SIGNIFICANCE: Surgical treatment of intestinal intussusception offers an excellent prognosis, but the potential life-threatening complications should be considered.

Outcome after surgical and conservative treatments of canine peritoneopericardial diaphragmatic hernia: A multi-institutional study of 128 dogs.

OBJECTIVE: To compare demographics and disease characteristics in dogs in which peritoneopericardial diaphragmatic hernia (PPDH) had been diagnosed and report outcomes after surgical treatment (ST) or conservative treatment (CT). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: One hundred twenty-eight dogs (91 ST, 37 CT) in which PPDH had been diagnosed. METHODS: Medical records were reviewed for demographics, perioperative findings, and outcomes. Follow-up was obtained via telephone interview and email correspondence with owners and referring veterinarians. Baseline variables were compared between treatment groups. RESULTS: Dogs treated surgically were younger (P < .001), more likely to be sexually intact (P = .002), more likely to have clinical signs from PPDH vs an incidental diagnosis (P < .001), and more likely to have other congenital abnormalities (P = .003) compared with dogs treated conservatively. Ninety-seven percent of ST dogs were discharged from hospitals. Intraoperative and postoperative complications were reported in 22% and 41% of dogs, respectively, although most complications were classified as low grade (75% and 83%, respectively). Follow-up was available in 87 dogs, at a median of 1062 days. Hernia recurrence was not reported in any surgically treated dog. The deaths of nine dogs (five ST, four CT) could be attributed to PPDH, and long median survival times were observed in both the ST and CT groups (8.2 and 5 years, respectively). CONCLUSION: Preoperative characteristics differed between dogs treated conservatively vs surgically. Surgical treatment was associated with low operative mortality, and both ST and CT dogs had good long-term survival. CLINICAL SIGNIFICANCE: A diagnosis of PPDH can confer a good long-term prognosis for both ST and CT dogs.

Heterogeneous photocatalysis in flow chemical reactors.

The synergy between photocatalysis and continuous flow chemical reactors has shifted the paradigms of photochemistry, opening new avenues of research with safer and scalable processes that can be readily implemented in academia and industry. Current state-of-the-art photocatalysts are homogeneous transition metal complexes that have favourable photophysical properties, wide electrochemical redox potentials, and photostability. However, these photocatalysts present serious drawbacks, such as toxicity, limited availability, and the overall cost of rare transition metal elements. This reduces their long-term viability, especially at an industrial scale. Heterogeneous photocatalysts (HPCats) are an attractive alternative, as the requirement for the separation and purification is largely removed, but typically at the cost of efficiency. Flow chemical reactors can, to a large extent, mitigate the loss in efficiency through reactor designs that enhance mass transport and irradiation. Herein, we review some important developments of heterogeneous photocatalytic materials and their application in flow reactors for sustainable organic synthesis. Further, the application of continuous flow heterogeneous photocatalysis in environmental remediation is briefly discussed to present some interesting reactor designs that could be implemented to enhance organic synthesis.

Perioperative characteristics, histological diagnosis, and outcome in cats undergoing surgical treatment of primary hyperparathyroidism.

OBJECTIVE: To report perioperative characteristics, complications, histopathologic diagnosis and outcome in cats undergoing surgical treatment for primary hyperparathyroidism (PHPT). STUDY DESIGN: Multi-institutional, retrospective case series. ANIMALS: Thirty-two client-owned cats. METHODS: Medical records of cats treated with surgical removal of 1 or more parathyroid gland(s) with confirmed histopathologic evaluation were reviewed. Cats were divided into preoperative ionized calcium (iCa) groups corresponding to the 33rd, 67th, and 100th percentiles of the preoperative iCa results of the study population. Follow-up consisted of phone conversation with owners or primary veterinarian. RESULTS: Ionized calcium was above reference range in all cats (median 1.8 mmol/L [interquartile range, 1.5-1.9]). Abnormal tissue was excised after cervical exploration in all cats. The most common histopathologic diagnoses were parathyroid adenoma in 20 of 32 (62.5%) cats and parathyroid carcinoma in 7 of 32 (21.9%) cats. At discharge, 6 of 32 (18.8%) cats had hypercalcemia, 5 of 32 (15.6%) had hypocalcemia, and 21 of 32 (65.6%) were normocalcemic. Preoperative iCa did not correlate with postoperative iCa. The median follow-up time was 332 days (range, 7-3156). Overall median survival time was 1109 days (95% CI, 856-1332). Survival time was not associated with preoperative iCa group, hypocalcemia at discharge, hypercalcemia at discharge, or diagnosis of carcinoma. CONCLUSION: In this cohort of cats, parathyroid adenoma was the most common cause of PHPT, and surgical treatment resulted in very good median survival time. Preoperative iCa was not predictive of postoperative hypocalcemia. CLINICAL SIGNIFICANCE: Surgical parathyroidectomy for treatment of PHPT in cats provides a favorable prognosis.

Synthesis of a Novel Type of 2,3'-BIMs via Platinum-Catalysed Reaction of Indolylallenes with Indoles.

Optimisation, scope and mechanism of the platinum-catalysed addition of indoles to indolylallenes is reported here to give 2,3'-BIMs with a novel core structure very relevant for pharmaceutical industry. The reaction is modulated by the electronic properties of the substituents on both indoles, with the 2,3'-BIMs favoured when electron donating groups are present. Although simple at first, a complex mechanism has been uncovered that explains the different behaviour of these systems with platinum when compared with other metals (e.g. gold). Detailed labelling studies have shown Pt-catalysed 6-endo-trig cyclisation of the indollylallene as the first step of the reaction and the involvement of two cyclic vinyl-platinum intermediates in equilibrium through a platinum carbene, as the key intermediates of the catalytic cycle towards the second nucleophilic attack and formation of the BIMs.

Development of autotaxin inhibitors: A series of zinc binding triazoles.

A series of inhibitors of Autotaxin (ATX) has been developed using the binding mode of known inhibitor, PF-8380, as a template. Replacement of the benzoxazolone with a triazole zinc-binding motif reduced crystallinity and improved solubility relative to PF-8380. Modification of the linker region removed hERG activity and led to compound 12 - a selective, high affinity, orally-bioavailable inhibitor of ATX. Compound 12 concentration-dependently inhibits autotaxin and formation of LPA in vivo, as shown in pharmacokinetic-pharmacodynamic experiments.

Structure-guided design of a selective inhibitor of the methyltransferase KMT9 with cellular activity.

Inhibition of epigenetic regulators by small molecules is an attractive strategy for cancer treatment. Recently, we characterised the role of lysine methyltransferase 9 (KMT9) in prostate, lung, and colon cancer. Our observation that the enzymatic activity was required for tumour cell proliferation identified KMT9 as a potential therapeutic target. Here, we report the development of a potent and selective KMT9 inhibitor (compound 4, KMI169) with cellular activity through structure-based drug design. KMI169 functions as a bi-substrate inhibitor targeting the SAM and substrate binding pockets of KMT9 and exhibits high potency, selectivity, and cellular target engagement. KMT9 inhibition selectively downregulates target genes involved in cell cycle regulation and impairs proliferation of tumours cells including castration- and enzalutamide-resistant prostate cancer cells. KMI169 represents a valuable tool to probe cellular KMT9 functions and paves the way for the development of clinical candidate inhibitors as therapeutic options to treat malignancies such as therapy-resistant prostate cancer.

Discovery of HC-7366: An Orally Bioavailable and Efficacious GCN2 Kinase Activator.

A series of activators of GCN2 (general control nonderepressible 2) kinase have been developed, leading to HC-7366, which has entered the clinic as an antitumor therapy. Optimization resulted in improved permeability compared to that of the original indazole hinge binding scaffold, while maintaining potency at GCN2 and selectivity over PERK (protein kinase RNA-like endoplasmic reticulum kinase). The improved ADME properties of this series led to robust in vivo compound exposure in both rats and mice, allowing HC-7366 to be dosed in xenograft models, demonstrating that activation of the GCN2 pathway by this compound leads to tumor growth inhibition.

FcRγ promotes T cell apoptosis in Fas-deficient mice.

Deficiency of Fas or its ligand leads to lymphocyte accumulation, lymphadenopathy, splenomegaly, and autoimmunity in mice and humans. Although the Fas pathway is important for limiting the number of peripheral T cells, inactivation of other pro-apoptotic molecules can also perturb T cell homeostasis independently of and/or in concert with Fas deficiency. Here, we show that combined deficiency of Fas and the Fc receptor common γ signaling chain (FcRγ) results in worsened T cell accumulation in comparison with mice lacking Fas alone, with a particularly marked increase in the number of TCRαß(+)CD4(-)CD8(-) double negative (DN) T cells. LPR FcRγ(-/-) mice exhibited reduced survival due to progressive lymphadenopathy. We further investigated the mechanisms whereby FcRγ deficiency promotes lymphoproliferative disease in Fas-mutant mice. Interestingly, there were no significant differences in T cell proliferation between LPR FcRγ(+/+) and LPR FcRγ(-/-) mice in vivo and in vitro. However, FcRγ deletion resulted in significantly decreased peripheral T cell apoptosis. Importantly, the observed increase in apoptosis was restricted to a subset of FcRγ(+) T cells. These T cells, but not those lacking FcRγ expression, exhibited increased activation of caspases 3 and 9, indicating a role for FcRγ in driving their apoptosis. FcRγ(+) DN T cells also showed enhanced sensitivity to TCR restimulation-induced cell death (RICD) despite lacking Fas, suggesting that TCR stimulation of autoreactive T cells in vivo may serve to eliminate FcRγ(+) T cells and thus exert partial control over lymphoproliferative disease. Hence, our data reveal a novel role for FcRγ in promoting peripheral T cell apoptosis in Fas-deficient mice, which may be of significant value in understanding autoimmune lymphoproliferative syndromes.

Educational Policy Development as a Leadership Experience: Inclusive Institutional Change.

Medical students have a unique opportunity to advocate for educational policies that promote best practices in undergraduate medical education. At the Geisel School of Medicine at Dartmouth, students play a crucial role in the development of medical education policies. This article describes two innovative, inclusive, and effective approaches to increase student engagement: (1) restructuring Medical Education Subcommittees to diversify student perspectives and (2) including students in a values-based design thinking approach to the development of new academic advancement and promotion and conduct policies. Through deliberate participation, medical students gain valuable skills that can be applied as future educators and academic leaders.

Four-dimensional electrical resistivity imaging for monitoring pumping-induced saltwater intrusion in a coastal aquifer.

Conventional views of saltwater intrusion (SWI), where a basal saline wedge extends inland below fresh groundwater, can be complicated by the influence of saltwater cells in the upper part of aquifers in areas affected by tidal cycles. Distinguishing the contribution of each saltwater source may prove fundamental for well design and resource management. Application of time-lapse electrical resistivity imaging (ERI) during a 32-h pumping test in a pristine unconfined coastal sand aquifer, affected by strong tidal ranges (>2 m), aimed to evaluate the potential of the method to characterize the source of induced SWI in four dimensions (three dimensions and time). Water level monitoring during the test revealed that at the end of pumping, the upper 2 m of the aquifer had dewatered in the vicinity of the well field, reversing hydraulic gradients between the aquifer and the sea. This induced SI, with mixing models of well head water quality suggesting that saline water contributions to total discharge rose from 4 % to 8 %. ERI results reflected dewatering through an increase in resistivity in the upper 2-6 m of the aquifer, while a decline in resistivity, relative to background conditions, occurred immediately below this, reflecting the migration of saline water through the upper layers of the aquifer to the pumping well. By contrast no change in resistivity occurred at depth, indicating no significant change in contribution from the basal saline water to discharge. Test findings suggest that future water resource development at the site should focus on close monitoring of shallow pumping, or pumping from deeper parts of the aquifer, while more generally demonstrating the value of time-lapse geophysical methods in informing coastal water resource management.

The Photochemical Mediated Ring Contraction of 4H-1,2,6-Thiadiazines To Afford 1,2,5-Thiadiazol-3(2H)-one 1-Oxides.

1,2,6-Thiadiazines treated with visible light and 3O2 under ambient conditions are converted into difficult-to-access 1,2,5-thiadiazole 1-oxides (35 examples, yields of 39-100%). Experimental and theoretical studies reveal that 1,2,6-thiadiazines act as triplet photosensitizers that produce 1O2 and then undergo a chemoselective [3 + 2] cycloaddition to give an endoperoxide that ring contracts with selective carbon atom excision and complete atom economy. The reaction was optimized under both batch and continuous-flow conditions and is also efficient in green solvents.

Transaminases Provide Key Chiral Building Blocks for the Synthesis of Selective M1/M4 Agonists.

We have developed a chiral route toward the synthesis of muscarinic M4 agonists that was enabled by the biocatalytic synthesis of the key spirocyclic diamine building blocks 10 and 12. Using these bifunctional compounds we were able to optimize a synthetic sequence toward a collection of advanced intermediates for further elaboration. These advanced intermediates were then used as starting points for early medicinal chemistry and the identification of selective M1/M4 agonists.

Safe, efficient, and reproducible gene therapy of the brain in the dog models of Sanfilippo and Hurler syndromes.

Recent trials in patients with neurodegenerative diseases documented the safety of gene therapy based on adeno-associated virus (AAV) vectors deposited into the brain. Inborn errors of the metabolism are the most frequent causes of neurodegeneration in pre-adulthood. In Sanfilippo syndrome, a lysosomal storage disease in which heparan sulfate oligosaccharides accumulate, the onset of clinical manifestation is before 5 years. Studies in the mouse model showed that gene therapy providing the missing enzyme α-N-acetyl-glucosaminidase to brain cells prevents neurodegeneration and improves behavior. We now document safety and efficacy in affected dogs. Animals received eight deposits of a serotype 5 AAV vector, including vector prepared in insect Sf9 cells. As shown previously in dogs with the closely related Hurler syndrome, immunosuppression was necessary to prevent neuroinflammation and elimination of transduced cells. In immunosuppressed dogs, vector was efficiently delivered throughout the brain, induced α-N-acetyl-glucosaminidase production, cleared stored compounds and storage lesions. The suitability of the procedure for clinical application was further assessed in Hurler dogs, providing information on reproducibility, tolerance, appropriate vector type and dosage, and optimal age for treatment in a total number of 25 treated dogs. Results strongly support projects of human trials aimed at assessing this treatment in Sanfilippo syndrome.

Clinical validation of a next-generation sequencing-based multi-cancer early detection "liquid biopsy" blood test in over 1,000 dogs using an independent testing set: The CANcer Detection in Dogs (CANDiD) study.

Cancer is the leading cause of death in dogs, yet there are no established screening paradigms for early detection. Liquid biopsy methods that interrogate cancer-derived genomic alterations in cell-free DNA in blood are being adopted for multi-cancer early detection in human medicine and are now available for veterinary use. The CANcer Detection in Dogs (CANDiD) study is an international, multi-center clinical study designed to validate the performance of a novel multi-cancer early detection "liquid biopsy" test developed for noninvasive detection and characterization of cancer in dogs using next-generation sequencing (NGS) of blood-derived DNA; study results are reported here. In total, 1,358 cancer-diagnosed and presumably cancer-free dogs were enrolled in the study, representing the range of breeds, weights, ages, and cancer types seen in routine clinical practice; 1,100 subjects met inclusion criteria for analysis and were used in the validation of the test. Overall, the liquid biopsy test demonstrated a 54.7% (95% CI: 49.3-60.0%) sensitivity and a 98.5% (95% CI: 97.0-99.3%) specificity. For three of the most aggressive canine cancers (lymphoma, hemangiosarcoma, osteosarcoma), the detection rate was 85.4% (95% CI: 78.4-90.9%); and for eight of the most common canine cancers (lymphoma, hemangiosarcoma, osteosarcoma, soft tissue sarcoma, mast cell tumor, mammary gland carcinoma, anal sac adenocarcinoma, malignant melanoma), the detection rate was 61.9% (95% CI: 55.3-68.1%). The test detected cancer signal in patients representing 30 distinct cancer types and provided a Cancer Signal Origin prediction for a subset of patients with hematological malignancies. Furthermore, the test accurately detected cancer signal in four presumably cancer-free subjects before the onset of clinical signs, further supporting the utility of liquid biopsy as an early detection test. Taken together, these findings demonstrate that NGS-based liquid biopsy can offer a novel option for noninvasive multi-cancer detection in dogs.