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The Hh/GLI signaling pathway was originally discovered in Drosophila as a major regulator of segment patterning in development. This pathway consists of a series of ligands (Shh, Ihh, and Dhh), transmembrane receptors (Ptch1 and Ptch2), transcription factors (GLI1–3), and signaling regulators (SMO, HHIP, SUFU, PKA, CK1, GSK3β, etc.) that work in concert to repress (Ptch1, Ptch2, SUFU, PKA, CK1, GSK3β) or activate (Shh, Ihh, Dhh, SMO, GLI1–3) the signaling cascade. Not long after the initial discovery, dysregulation of the Hh/GLI signaling pathway was implicated in human disease. Activation of this signaling pathway is observed in many types of cancer, including basal cell carcinoma, medulloblastoma, colorectal, prostate, pancreatic, and many more. Most often, the activation of the Hh/GLI pathway in cancer occurs through a ligand-independent mechanism. However, in benign disease, this activation is mostly ligand-dependent. The upstream signaling component of the receptor complex, SMO, is bypassed, and the GLI family of transcription factors can be activated regardless of ligand binding. Additional mechanisms of pathway activation exist whereby the entirety of the downstream signaling pathway is bypassed, and PTCH1 promotes cell cycle progression and prevents caspase-mediated apoptosis. Throughout this review, we summarize each component of the signaling cascade, non-canonical modes of pathway activation, and the implications in human disease, including cancer.

With rapid advances in our understanding of cancer, there is an expanding number of potential novel combination therapies, including novel–novel combinations. Identifying which combinations are appropriate and in which subpopulations are among the most difficult questions in medical research. We conducted a Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)-guided systematic review of trials of novel–novel combination therapies involving immunotherapies or molecular targeted therapies in advanced solid tumors. A MEDLINE search was conducted using a modified Cochrane Highly Sensitive Search Strategy for published clinical trials between July 1, 2017, and June 30, 2020, in the top-ranked medical and oncology journals. Trials were evaluated according to a criterion adapted from previously published Food and Drug Administration guidance and other key considerations in designing trials of combinations. This included the presence of a strong biological rationale, the use of a new established or emerging predictive biomarker prospectively incorporated into the clinical trial design, appropriate comparator arms of monotherapy or supportive external data sources and a primary endpoint demonstrating a clinically meaningful benefit. Of 32 identified trials, there were 11 (34%) trials of the novel–novel combination of anti-programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1) and anti-cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) therapy, and 10 (31%) trials of anti-PD-1/PD-L1 and anti-vascular endothelial growth factor (VEGF) combination therapy. 20 (62.5%) trials were phase II trials, while 12 (37.5%) were phase III trials. Most (72%) trials lacked significant preclinical evidence supporting the development of the combination in the given indication. A majority of trials (69%) were conducted in biomarker unselected populations or used pre-existing biomarkers within the given indication for patient selection. Most studies (66%) were considered to have appropriate comparator arms or had supportive external data sources such as prior studies of monotherapy. All studies were evaluated as selecting a clinically meaningful primary endpoint. In conclusion, designing trials to evaluate novel–novel combination therapies presents numerous challenges to demonstrate efficacy in a comprehensive manner. A greater understanding of biological rationale for combinations and incorporating predictive biomarkers may improve effective evaluation of combination therapies. Innovative statistical methods and increasing use of external data to support combination approaches are potential strategies that may improve the efficiency of trial design. Designing trials to evaluate novel–novel combination therapies presents numerous challenges to demonstrate efficacy in a comprehensive manner. A greater understanding of biological rationale for combinations and incorporating predictive biomarkers may improve effective evaluation of combination therapies. Innovative statistical methods and increasing use of external data to support combination approaches are potential strategies that may improve the efficiency of trial design.

Background: Telestroke networks have effectively increased the number of ischemic stroke patients who have access to acute stroke therapy. However, the availability of a dedicated group of stroke subspecialists is not always feasible. Hypothesis: Rates of tPA recommendation, accuracy of final diagnosis and post-tPA hemorrhagic complications do not differ significantly between neurologists and an emergency-medicine physician during telestroke consultations. Methods: Retrospective review of all telestroke consults performed at a comprehensive stroke center during one year. Statistical analysis: Chi squared test. Results: 303 consults were performed among 6 spoke sites. 16% (48/303) were completed by the emergency medicine physician; 25% (76/303) were performed by non stroke-trained neurologists, and 60% (179/303) were completed by a board-certified Vascular Neurologist. Overall rate of tPA recommendation was 40% (104/255), 38% (18/48), 41% (73/179), and 41% (31/76) among the all neurology-trained, emergency medicine- trained, stroke neurology-trained and other neurology- trained provider groups respectively (p = .427). Accuracy of final stroke diagnosis was 77% (14/18) and 72% (75/104) in the emergency-medicine trained and neurology-trained provider groups (p = 0.777) No symptomatic hemorrhagic complications following the administration of tPA via telestroke consultation occurred in any group over this time period. One asymptomatic intracerebral hemorrhage was observed (0.96% or 1/104) in the neurology-trained provider group. Conclusion: Our results did not illustrate any statistically significant difference between care provided by an Emergency Medicine-trained physician and neurologists during telestroke consultation. While our study is limited by its relatively low numbers, it suggests that identifying a non-neurologist provider who has requisite clinical experience with acute stroke patients can safely and appropriately provide telestroke consultation. The lack of formerly trained neurologists, therefore may not need to serve as an impediment to building an effective telestroke network. Future efforts should be focused on illuminating all strategies that facilitate sustainable telestroke implementation.

Background Sedentary time has been shown to have deleterious effects on health. Today's workforce spends most of the day sitting. The objective of this study was to evaluate the effect of workplace interventions to decrease sedentary time on cardiometabolic risk factors. Methods Sedentary office workers (n=127; ages 22‐64; BMI=28.5±6.1 kg/m 2 ) were recruited from three Minnesota employers and randomized to one of four intervention groups for 6 months: 1) Control, 2) Move (蠅30 min of light activity during the workday), 3) Stand (standing 蠅50% of the workday using a sit‐stand workstation), or 4) Stand+Move (combined Stand and Move). A 'metabolic risk score' (MRS) was calculated at baseline and 6 months using the following cardiometabolic risk factors: blood pressure, fasting blood glucose, log of fasting triglycerides, and HDL‐cholesterol. A z‐score was computed for each risk factor for each subject [(subject value – sample mean)/sample SD]. The MRS was derived by summing the z‐scores (HDL‐z subtracted). Results There was a significant intervention effect on MRS (p<.01). A seasonal time trend was observed, with MRS increasing significantly in Control and Move groups (p<.001), but not for the Stand and Stand+Move. Results (mean±se) are shown in Figure 1. Conclusions Sit‐stand workstations combined with an intervention to increase light physical activity at work may be an effective tool to improve cardiometabolic risk. image

Physical resilience, defined as the ability to resist and respond to challenges and return to homeostasis, has been postulated to be a determinant of healthspan and lifespan. To test this premise, we have begun to optimize a standardized, clinically relevant battery of resilience challenges, including anesthesia and cisplatin chemotherapy, and recovery outcome measures in younger (2–3 month) and older (26–29 month) mice. We have also started to test whether interventions shown to increase murine lifespan‐acarbose, rapamycin, and 17a‐estradiol‐impact the resilience of older mice. Our preliminary data demonstrate that younger mice (n = 10) have faster dorsal to sternal recumbence recovery time compared to older mice (n = 8) (39 ± 1 sec vs. 67 ± 6 sec, p < 0.001) following a 5‐minute 4% isoflurane anesthesia challenge. In response to a 5‐day cisplatin (2 mg/kg/day) challenge, both younger (n=5) and older (n=4) mice lost a significant percent of their baseline body weight (younger mice −10.6 ± 0.8% and older mice −13.2 ± 2.1%, p = 0.26), however, older mice failed to regain body weight 11 days after the challenge (younger mice −0.9 ± 0.8% and older mice −9.8 ± 0.8%, p < 0.01). Changes in body weight in younger and older mice were largely attributable to the loss/regain of fat mass. Interestingly, 8 weeks of either acarbose (1,000 ppm) or 17α‐estradiol (14.4 ppm) administration to older female mice improved body weight recovery after the cisplatin challenge, while rapamycin (14 ppm) had no appreciable effect. Our preliminary results demonstrate that younger mice, as expected, have greater resilience than older mice to clinically relevant physical challenges. The age at which physical resilience starts to decline requires further investigation. Our early observations suggest interventions that extend health‐ and lifespan may impact resilience in older mice. Ultimately, a standardized battery of resilience challenges may provide a new framework for testing and translating interventions that target the fundamental biology of aging. Support or Funding Information This work was supported by grant AG52958 from the National Institutes of Health. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Two out of every thousand newborns are born deaf or hard of hearing. Sixty percent of hearing loss has an identifiable genetic cause. One of the most common forms of genetic hearing loss is Pendred syndrome. Disease causing genetic alterations or pathogenic variants in SLC26A4 are associated with Pendred Syndrome, a disorder characterized by bilateral sensorineural hearing loss and occasionally thyroid goiter. Patients with SLC26A4 pathogenic variants have an enlarged vestibular aqueduct, a malformation of the temporal bone. Pendrin, the protein product of SLC26A4 , is a solute carrier. Pendrin is expressed in the inner ear, thyroid, and kidney. In the inner ear, pendrin modulates the transport of bicarbonate and chloride, allowing bicarbonate to leave the supporting cells of the inner ear and chloride to enter. This helps maintain a relatively neutral pH environment in the inner ear. It is not known how the abnormal SLC26A4 protein products modulate hearing loss and contribute to temporal bone abnormalities. The goal of this study was to model SLC26A4 associated genetic hearing loss in zebrafish and elucidate a mechanism responsible for hearing loss in patients with pathogenic variants in SLC26A4 . To model the human condition, slc26a4 mutations in zebrafish were generated via transcription activator like effector nucleases (TALENs) at the site of the the most common SLC26A4 pathogenic variants. We obtained a 23bp deletion mutation in exon 10 of slc26a4 . We hypothesized that fish with biallelic slc26a4 mutations would closely recapitulate the human phenotype and have significant hearing loss and structural ear malformations. To test for deafness in zebrafish, we developed a hearing assessment which exposes larval or adult fish to specific sound stimuli while recording startle responses. To quantitatively study the slc26a4 mutant ear phenotype, we took measurements of the posterior and anterior otolith area and the ear area using bright field microscopy. Results indicate that homozygous embryos tested at 5 days post fertilization, the day of hearing onset in zebrafish, have hearing that was not different that wildype, however adult fish with two copies of mutated slc26a4 have frequency specific reduced responses at 400 Hz, 80db. These results indicate that slc26a4 mutants have progressive frequency specific hearing loss. Bright field imaging and measurement analysis showed that slc26a4 mutant larvae have significantly smaller standardized posterior otolith areas, anterior otolith areas, and ear areas on average compared to wildtype at 5dpf. This closely matches the human phenotype, as humans with SLC26A4 pathogenic variants may have progressive hearing loss and temporal bone abnormalities. Support or Funding Information Mayo Center for Individualized Medicine Mayo Clinic Department of Otorhinolaryngology NIH 5R25GM075148‐11 This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Mayo Clinic annually trains over 1000 predoctoral students and postdoctoral research fellows in medicine and biomedical sciences. However, training opportunities are limited for those interested in teaching and mentoring methodologies. Therefore, Mayo Clinic predoctoral students sought to provide trainee‐led learning opportunities for fellow trainees. A survey was sent to all Mayo Clinic predoctoral and postdoctoral trainees to determine preferences for pedagogy education formats. In response to survey results, the Twin Talk Series was developed. A Twin Talk is a set of one‐hour talks comprised of a Take‐in session and one or more Try‐out session(s). At the Take‐in session, a content expert discusses a pedagogical technique, and at the following Try‐out session(s), trainee volunteers practice the technique among themselves. Eight Twin Talks have been organized to date on several teaching techniques and pedagogical development topics. The impact of each Twin Talk, including understanding, format, and interest level, was evaluated through survey tools. Survey results suggested trainees favored in‐person pedagogy expert presenters during the Take‐in session over video conferences. Furthermore, modeling of the teaching technique by the expert presenter during the Take‐in session was well‐received. However, participation in the follow‐up Try‐out practice session was limited. Overall, the Twin Talk Series is a promising method for trainee‐led, peer‐to‐peer pedagogy education, particularly at institutions with limited access to pedagogical resources. The Twin Talk Series will continue to be refined to increase trainee participation, particularly in the follow‐up Try‐out sessions, and to provide relevant opportunities for trainees to practice teaching. Support or Funding Information Mayo Foundation of Medical Education and Research This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Mono‐and polyunsaturated fats in nuts, such as pistachios (PI), may improve lipoprotein and glycemic status. Overweight persons trend for dyslipidemia and high diabetes risk. The lower limits of PI consumption required for health benefits are unknown. This study determined if PI at bedtime promotes beneficial changes in metabolic status in overweight persons. Overweight subjects (n=22; 55 ± 2 yrs; 90 ± 3.6 kg; BMI 31 ± 1) were randomized to 35.4 g PI self‐administered at bedtime or control (CO; no PI) for 6 weeks. There was no difference in physical activity level between subjects receiving PI and CO. Body weight was not different in PI and CO at wk 0, 1, 2, 4, and 6. HDL cholesterol at wk 0 and 6 in PI was 49.2 ± 3.5 to 50.3 ± 3.0, and CO was 60.8 ± 3.8 and 58.0 ± 2.7 mg/dL, at wk 6 PI had improved slightly relative to CO (P= 0.12). Plasma glucose at wk 0 and 6 in PI was 104.6 ± 2.8 and 99.6 ± 2.6 mg/dL, and CO was 102.0 ± 2.8 and 101.1 ± 2.6 mg/dL, at wk 6 PI had improved slightly relative to CO (P= 0.16). Plasma insulin at wk 0 and 6 in PI was 8.5 ± 1.0 and 8.4 ± 0.9, and CO was 7.2 ± 0.7 to 8.8 ± 0.7 μU/mL, at wk 6 PI had improved slightly relative to CO (P = 0.21). Beneficial changes in HDL, glucose, and insulin probably requires slightly more than 35.4 g/d PI for 6 wks. To increase the probability of metabolic health benefits, nutritionists may wish to encourage levels of daily PI consumption higher than those administered in this study.