Beluga whale and bottlenose dolphin ACE2 proteins allow cell entry mediated by spike protein from three variants of SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viruses infect numerous non-human species. Spillover of SARS-CoV-2 into novel animal reservoirs may present a danger to host individuals of these species, particularly worrisome in populations already endangered or threatened by extinction. In addition, emergence in new reservoirs could pose spillback threats to humans, especially in the form of virus variants that further mutate when infecting other animal hosts. Previous work suggests beluga whales (Delphinapterus leucas) and bottlenose dolphins (Tursiops truncatus) may be at risk owing to their formation of social groups, contact with humans, exposure to contaminated wastewater, and structure of their angiotensin-converting enzyme 2 (ACE2) proteins, which SARS-CoV-2 uses as a cellular receptor. We examined marine-mammal susceptibility to virus infection by challenging 293T cells expressing beluga or dolphin ACE2 with pseudovirions bearing the SARS-CoV-2 spike protein. Beluga and dolphin ACE2 were sufficient to allow cell entry by an early pandemic isolate (Wuhan-Hu-1) and two evolved variants (Delta B.1.617.2 and Omicron BA.1 strains). We conclude that SARS-CoV-2 poses a potential threat to marine mammal reservoirs that should be considered in surveillance efforts.

Influence of season, age, sex, and time of day on the endocrine profile of the common bottlenose dolphin (Tursiops truncatus).

Understanding baseline hormone levels, the magnitude of intra-individual variability, and their variation as a function of life history is difficult in toothed whales (e.g. dolphins and porpoises) because of the effects of capture stress. To determine the endocrine profile of the common bottlenose dolphin (Tursiops truncatus) as a function of season, time of day (TOD), age, sex, and reproductive status, blood corticosteroids, thyroid hormones, and catecholamines were repeatedly measured in a managed-care population exposed to ambient light and water temperatures of San Diego Bay. Additionally, fecal hormone metabolites were assessed for cortisol, aldosterone, and triiodothyronine. Samples were collected at two to four-week intervals over a period of two years, and multiple times within a day at monthly intervals over a year. Samples were collected through the voluntary participation of the dolphins in the blood draws and fecal collections in order to avoid the effects of handling stress. All serum hormones except aldosterone significantly varied with season and all serum hormones except total thyroxine significantly varied as a function of TOD. Fecal glucocorticoid metabolites significantly correlated with circulating cortisol levels, and there was a significant seasonal effect on triiodothyronine fecal metabolites. Strong seasonal effects demonstrated complex interactions with age and sex suggesting that contextual information is critical to interpreting differences in endocrine profiles. Strong circadian patterns further suggest that sampling design is important to the interpretation of blood or fecal collections, particularly since diurnal changes in some serum hormone levels are similar to the magnitude of seasonal differences. Despite potential impacts of feeding schedules on diurnal patterns, managed care populations can provide important insights into seasonal and age-related endocrine changes in toothed whales.

Sodium selenate treatment mitigates reduction of bone volume following traumatic brain injury in rats.

OBJECTIVES: Administration of sodium selenate to rats given traumatic brain injury (TBI) attenuates brain damage and improves long-term behavioural outcomes. We have previously provided evidence that TBI causes bone loss in rats, however the effect of sodium selenate treatment on bone quantity following TBI is unknown. METHODS: Rats were randomly assigned into sham injury or fluid percussion injury (FPI) groups and administered saline or sodium selenate for 12 weeks post-injury. Femora were analysed using histomorphometry, peripheral quantitative computed tomography (pQCT) and biomechanical testing. RESULTS: Distal metaphyseal trabecular bone volume fraction of FPI-selenate rats was higher than FPI-vehicle rats (41.8%; p<0.01), however, femora from selenate-treated groups were shorter in length (4.3%; p<0.01) and had increased growth plate width (22.1%; p<0.01), indicating that selenate impaired long bone growth. pQCT analysis demonstrated that distal metaphyseal cortical thickness was decreased in TBI rats compared to shams (11.7%; p<0.05), however selenate treatment to TBI animals offset this reduction (p<0.05). At the midshaft we observed no differences in biomechanical measures. CONCLUSION: These are the first findings to indicate that mitigating TBI-induced neuropathology may have the added benefit of preventing osteoporosis and associated fracture risk following TBI.

Forward shift of feeding buzz components of dolphins and belugas during associative learning reveals a likely connection to reward expectation, pleasure and brain dopamine activation.

For many years, we heard sounds associated with reward from dolphins and belugas. We named these pulsed sounds victory squeals (VS), as they remind us of a child's squeal of delight. Here we put these sounds in context with natural and learned behavior. Like bats, echolocating cetaceans produce feeding buzzes as they approach and catch prey. Unlike bats, cetaceans continue their feeding buzzes after prey capture and the after portion is what we call the VS. Prior to training (or conditioning), the VS comes after the fish reward; with repeated trials it moves to before the reward. During training, we use a whistle or other sound to signal a correct response by the animal. This sound signal, named a secondary reinforcer (SR), leads to the primary reinforcer, fish. Trainers usually name their whistle or other SR a bridge, as it bridges the time gap between the correct response and reward delivery. During learning, the SR becomes associated with reward and the VS comes after the SR rather than after the fish. By following the SR, the VS confirms that the animal expects a reward. Results of early brain stimulation work suggest to us that SR stimulates brain dopamine release, which leads to the VS. Although there are no direct studies of dopamine release in cetaceans, we found that the timing of our VS is consistent with a response after dopamine release. We compared trained vocal responses to auditory stimuli with VS responses to SR sounds. Auditory stimuli that did not signal reward resulted in faster responses by a mean of 151 ms for dolphins and 250 ms for belugas. In laboratory animals, there is a 100 to 200 ms delay for dopamine release. VS delay in our animals is similar and consistent with vocalization after dopamine release. Our novel observation suggests that the dopamine reward system is active in cetacean brains.

Single port (SP) robotic pyeloplasty in a pediatric patient.

We began using the da Vinci single port (SP) robot for pediatric urologic surgeries at our institution due to limited access to the multiport robot. Availability of this technology has allowed us to schedule cases in a timelier fashion and increase access to minimally invasive urologic surgery for children in our area. Here, we report our technique for transperitoneal SP robotic pyeloplasty in the case of a 7 year-old boy with left ureteropelvic junction obstruction. Our technique was refined over a series of 10 patients under the age of 18. Highlights of the SP RALP technique include one 3 cm, concealed incision over the pubic tubercle, gentle frog leg positioning and burping of the boom to create optimal angle for robotic docking, and use of a "floating dock" to obtain 10 cm distance from target anatomy which is essential in smaller pediatric patients. SP pyeloplasty is safe and feasible in children and offers a concealed single incision alternative to the multiport approach.

Bilateral proximal ureteral and ureterovesical junction obstruction in a child.

The most common sites of ureteral obstruction in children are at the level of the ureteropelvic junction (UPJ) and ureterovesical junction (UVJ). Bilateral hydronephrosis or hydroureteronephrosis due to varying degrees of obstruction at the UPJ or UVJ is common in children and typically improves with time. Clinically significant obstruction at both locations in an ipsilateral ureter occurs less commonly and rarely requires both dismembered pyeloplasty and ureteral reimplantation. We believe this case report is the first description of bilateral proximal and distal ureteral obstruction requiring both dismembered pyeloplasty and ureteral reimplantation.

Naphthoquinone-tyrptophan reduces neurotoxic Aß*56 levels and improves cognition in Alzheimer's disease animal model.

An increasing body of evidence indicates a role for oligomers of the amyloid-ß peptide (Aß) in the neurotoxicity of this peptide and the pathology of Alzheimer's disease (AD). Several neurotoxic oligomeric forms of Aß have been noted ranging from the larger Amyloid ß-Derived Diffusible Ligands (ADDLs) to smaller trimers and dimers of Aß. More recently a dodecameric form of Aß with a 56 kDa molecular weight, denoted Aß*56, was shown to cause memory impairment in AD model mice. Here, we present for the first time a potential therapeutic strategy for AD that targets the early stages in the formation of neurotoxic Aß*56 oligomers using a modified quinone-Tryptophan small molecule N-(3-chloro-1,4-dihydro-1,4-dioxo-2-naphthalenyl)-L-Tryptophan (Cl-NQTrp). Using NMR spectroscopy we show that this compound binds the aromatic recognition core of Aß and prevents the formation of oligomers. We assessed the effect of Cl-NQTrp in vivo in transgenic flies expressing Aß(1-42) in their nervous system. When these flies were fed with Cl-NQTrp a marked alleviation of their Aß-engendered reduced life span and defective locomotion was observed. Finally, intraperitoneal injection of Cl-NQTrp into an aggressive AD mouse model reduced the level of the Aß*56 species in their brain and reversed their cognitive defects. Further experiments should assess whether this is a direct effect of the drug in the brain or an indirect peripheral effect. This is the first demonstration that targeted reduction of Aß*56 results in amelioration of AD symptoms. This second generation of tryptophan-modified naphthoquinones could therefore serve as potent disease modifying therapeutic for AD.

Ovotesticular Disorder Mimicking Acute Spermatic Cord Torsion.

Ovotesticular disorder is a rare form of sexual development in which a patient may have one ovary and one testis, or more commonly a gonad or gonads containing both ovarian and testicular tissue. Patients with this condition typically present in infancy with ambiguous genitalia. Delayed presentations of clinically symptomatic, older patients with normal external genitalia are extremely rare. We present a case of a 14-year-old male with normal external genitalia who presented with symptoms and signs consistent with spermatic cord torsion but found to have ovotesticular disorder on evaluation.

A narrative review of the history and evidence-base for the timing of orchidopexy for cryptorchidism.

INTRODUCTION: Consensus recommendations for surgical management of cryptorchidism recommend orchidopexy between 6 and 18 months of age. The COVID-19 pandemic has impacted elective surgical scheduling. OBJECTIVE: In response to the COVID-19 pandemic, we sought to review the available data regarding the natural history, surgical management, and infertility- and cancer-related risks associated with cryptorchid testes. The purpose of this review is to provide parents, referring providers, and surgeons with information to inform their decisions to proceed with or delay orchidopexy. METHODS: A retrospective review and analysis of all available articles relevant to the natural history, surgical management, and infertility- and cancer-related risks of cryptorchidism present on PubMed, SCOPUS, and Cochrane Library was conducted. RESULTS: The quality of historic literature pertaining to the effect of cryptorchidism on fertility and malignancy differ, with poorer data available on fertility. Cryptorchid testes may show histologic differences as early as birth, and some of these changes may have prognostic value in future fertility. Formerly unilateral cryptorchid men have slightly but not significantly reduced paternity rates compared to the general population. Cryptorchid testes have an increased risk of germ cell carcinogenesis, and robust data suggest the risk for malignancy in cryptorchid testes increases substantially after puberty. CONCLUSION: The current body of evidence regarding the risks for future infertility and testicular cancer support the consensus recommendations for surgical correction of cryptorchidism between 6 and 18 months of age. During the uncertain time of the COVID-19 pandemic, decision for orchidopexy is a shared-decision between physician and parent. For an infant or young boy with a unilateral undescended testes, delaying orchidopexy several months until a time of decreased exposure risk is unlikely to result in substantial or sustained fertility or malignant risks.

Bilateral Calculi in a Child With Bilateral Single System Ureteroceles.

Ureteroceles in children are detected with prenatal sonography and less commonly during the evaluation following a urinary tract infection. Rarely do ureteroceles in the pediatric population present with stones, particularly in a bilateral fashion. We present a case of a 5-year-old boy found to have bilateral intravesical single system ureteroceles harboring multiple large calculi treated successfully with a staged endoscopic approach.

Impact of hospital transfer on testicular torsion outcomes: A systematic review and meta-analysis.

INTRODUCTION: Testicular torsion is an emergent condition requiring prompt treatment. Previous studies have suggested transfer of pediatric testicular torsion cases may be detrimental to patient outcomes. Findings have not reached statistical significance. No study has quantitatively analyzed all literature reporting outcomes for transferred torsion patients. The aim of this study was to elucidate the impact of hospital transfer on pediatric testicular torsion outcomes through a systematic review and meta-analysis. METHODS: A predefined study protocol registered with PROSPERO was developed according to PRISMA. A comprehensive literature review of articles investigating outcomes for pediatric testicular torsion for transferred and non-transferred (treated "directly" at presentation institution) patients with orchiectomy as the primary outcome was conducted by systematically searching PubMed and Embase. Potential studies were screened against a predefined study protocol. Meta-analysis using a random effects model with transferred status as the "intervention" was performed using Review Manager 5.3. RESULTS: Of 18 eligible studies, 9 retrospective studies comprised of 2564 patients (532 transferred and 2032 direct) were suitable for quantitative analysis. Main analysis showed transfer status does not have a significant effect on torsion outcomes (RR 0.96 [95% CI 0.78-1.17]; I2 = 44%). Subgroup analysis for torsion patients presenting within 24 h of symptom onset shows patients who are transferred to another facility for management are more likely to undergo orchiectomy than those treated at their presenting institution (RR 0.35 [95% CI 0.24-0.51]; I2 = 4%). CONCLUSIONS: In this meta-analysis, hospital transfer does not affect orchiectomy rate in pediatric patients with testicular torsion when pooling data from all presentation timeframes. Subgroup analysis of patients presenting with testicular torsion in an acute setting (<24 h of symptom onset) suggests the delay associated with hospital transfer has a deleterious effect on testicular viability.

Selective viral immunosuppression of the graft-versus-host reaction.

Graft-vs.-host (GVH) reactivity of parental lymph node (LN) cells was assayed by measurements of 3H-thymidine incorporation in vivo in spleens of irradiated F1 recipients. Preincubation of parental LN cells with vesicular stomatitis virus (VSV) for 2 h at 37 degrees C followed by washing resulted in an 85-90% reduction in splenic radioactivity, as did injection of VSV on days 0-2 after recipients received untreated parental LN cells. In contrast, 3H-thymidine incorporation in the spleens or irradiated F1 hosts was not affected by VSV when F1 bone marrow cells were incubated with the virus. In addition, preincubation of F1 B cells with VSV still allowed these syngeneic B cells to be recruited into proliferation by mitomycin-treated parental LN cells. The inhibitory effect of VSV, thus, seems to be specific for T-cell proliferation. These observations suggest that viral immunosuppression might be capable of being developed into a useful strategy for selective deletion of lymphocytes capable of reacting against histocompatibility antigens and initiating GVH reactions.

Endodermal origin of bladder trigone inferred from mesenchymal-epithelial interaction.

PURPOSE: In the classic view of bladder development the trigone originates from the mesoderm derived wolffian ducts while the remainder of the bladder originates from the endoderm derived urogenital sinus. Recent molecular developmental studies have questioned the veracity of this received wisdom, suggesting an endodermal origin for the trigone. To shed further light on this issue we observed mesenchymal-epithelial interactions between trigone epithelium and fetal urogenital sinus mesenchyma to infer the trigonal germ layer of origin. MATERIALS AND METHODS: Mouse trigone epithelium was recombined with fetal rat urogenital sinus mesenchyma in tissue recombinant grafts that were placed beneath the renal capsule of athymic mouse hosts. Grafts were harvested at 4 weeks. Control grafts with bladder dome and ureteral epithelium were also examined. Tissues were evaluated with hematoxylin and eosin, and Hoechst dye 33258 to confirm cell species origin. Immunohistochemistry was done with androgen receptor, broad spectrum uroplakin, dorsolateral prostate secretions and seminal vesicle secretions to differentiate prostatic and seminal vesicle differentiation. RESULTS: Grafts of mouse trigone epithelium with fetal rat urogenital sinus mesenchyma yielded epithelial tissue that stained for dorsolateral prostate secretions but not for seminal vesicle secretions. Control grafts of bladder dome epithelium yielded the expected endodermal prostate differentiation. Control grafts of ureteral epithelium yielded the expected mesodermal seminal vesicle differentiation. CONCLUSIONS: The consistent finding of prostatic epithelium in tissue recombinants of trigone epithelium and fetal urogenital sinus mesenchyma reinforces the hypothesis that the trigone is derived from the endoderm and not from the mesoderm, as commonly accepted.

Laparoscopic versus open pyeloplasty in children: preliminary report of a prospective randomized trial.

PURPOSE: Pyeloplasty for ureteropelvic junction obstruction in children has traditionally been performed using an open technique. Although laparoscopic pyeloplasty has been shown to be comparable and possibly superior to open pyeloplasty in adult studies, such results in the pediatric population are limited. We evaluated outcomes between transperitoneal laparoscopic and open pyeloplasty in children. MATERIALS AND METHODS: All children 1 to 18 years old with ureteropelvic junction obstruction requiring operative repair were offered enrollment in the study. Patients were prospectively randomized to either transperitoneal laparoscopic or open pyeloplasty through a flank incision. RESULTS: We reviewed 20 patients (mean age 7.8 years) who underwent laparoscopy and 19 (7.2 years) who underwent open surgery (p = 0.48). Mean followup was similar between the groups (laparoscopic 8.1 months vs open 11.1 months, p = 0.38). Mean operative time was 151 minutes (range 94 to 213) for laparoscopy and 130 minutes (83 to 225) for open surgery (p = 0.09). Mean hospitalization was 29.3 hours (range 20.5 to 48) for laparoscopy and 36.2 hours (24 to 73) for open surgery (p = 0.06). Analgesic usage was similar between the groups. One failure in the open arm required a revision. Operative, hospital, anesthetic and total charges were similar between the groups. CONCLUSIONS: Laparoscopic pyeloplasty appears to be a safe and effective alternative to open pyeloplasty in children. Although the cost is similar, there is a trend toward longer operative times in the laparoscopic group but a shorter overall hospitalization. As more patients are enrolled in the study, these differences may prove significant.

Large-scale comparison of four binding site detection algorithms.

A large-scale evaluation and comparison of four cavity detection algorithms was carried out. The algorithms SiteFinder, fpocket, PocketFinder, and SiteMap were evaluated on a protein test set containing 5416 protein-ligand complexes and 9900 apo forms, corresponding to a subset of the set used earlier for benchmarking the PocketFinder algorithm. For the holo structures, all four algorithms correctly identified a similar amount of pockets (around 95%). SiteFinder, using optimized parameters, SiteMap, and fpocket showed similar pocket ranking performance, which was defined by ranking the correct binding site on rank 1 of the predictions or within the first 5 ranks of the predictions. On the apo structures, PocketFinder especially and also SiteFinder (optimized parameters) performed best, identifying 96% and 84% of all binding sites, respectively. The fpocket program predicts binding sites most accurately among the algorithms evaluated here. SiteFinder needed an average calculation time of 1.6 s compared with 2 min for SiteMap and around 2 s for fpocket.