bluuebunny/crossref_metadata_2025 huggingface dataset:
Direct-to-Implant Versus 2-Stage Breast Reconstruction
Authors: Piper Merisa L. , Rios-Diaz Arturo J. , Kimia Rotem , Cunning Jessica , Broach Robyn , Wu Liza , Serletti Joseph , Fosnot Joshua
Published: June 2022

Background

Two-stage (TS) implant-based reconstruction is the most commonly performed method of reconstruction after mastectomy. A growing number of surgeons are offering patients direct-to-implant (DTI) reconstruction, which has the potential to minimize the number of surgeries needed and time to complete reconstruction, as well as improve health care utilization. However, there are conflicting data regarding the outcomes and complications of DTI, and studies comparing the 2 methods exclusively are lacking.

Methods

Patients undergoing implant-based reconstruction after mastectomy within a large interstate health system between 2015 and 2019 were retrospectively identified and grouped by reconstruction technique (TS and DTI). The primary outcomes were a composite of complications (surgical site occurrences), health care utilization (reoperations, unplanned emergency department visits, and readmissions), and time to reconstruction completion. Risk-adjusted logistic and generalized linear models were used to compare outcomes between TS and DTI.

Results

Of 104 patients, 42 underwent DTI (40.4%) and 62 underwent TS (59.6%) reconstruction. Most demographic characteristics, and oncologic and surgical details were comparable between groups (P > 0.05). However, patients undergoing TS reconstruction were more likely to be publicly insured, have a smoking history, and undergo skin-sparing instead of nipple-sparing mastectomy. The composite outcome of complications, reoperations, and health care utilization was higher for DTI reconstruction within univariate (81.0% vs 59.7%, P = 0.03) and risk-adjusted analyses (odds ratio, 3.78 [95% confidence interval [CI], 1.09–13.9]; P < 0.04). Individual outcome assessment showed increased mastectomy flap necrosis (16.7% vs 1.6%, P < 0.01) and reoperations due to a complication (33.3% vs 16.1%; P = 0.04) in the DTI cohort. Although DTI patients completed their aesthetic revisions sooner than TS patients (median, 256 days vs 479 [P < 0.01]; predicted mean difference for TS [reference DTI], 298 days [95% CI, 71–525 days]; P < 0.01), the time to complete reconstruction (first to last surgery) did not differ between groups (median days, DTI vs TS, 173 vs 146 [P = 0.25]; predicted mean difference [reference, DTI], −98 days [95% CI, −222 to 25.14 days]; P = 0.11).

Conclusions

In this cohort of patients, DTI reconstruction was associated with higher complications, reoperations, and health care utilization with no difference in time to complete reconstruction compared with TS reconstruction. Further studies are warranted to investigate patient-reported outcomes and cost analysis between TS and DTI reconstruction.
A Transcriptomic Liquid Biopsy Assay for Predicting Resistance to Neoadjuvant Therapy in Esophageal Squamous Cell Carcinoma
Authors: Okuno Keisuke , Tokunaga Masanori , Kinugasa Yusuke , Baba Hideo , Kodera Yasuhiro , Goel Ajay
Published: June 2022

Objective:

The aim of this study was to establish a liquid-biopsy assay to predict response to neoadjuvant therapy (NAT) in esophageal squamous cell carcinoma (ESCC) patients.

Summary Background Data:

Pretreatment prediction of resistance to NAT is of great significance for the selection of treatment options in ESCC patients. In this study, we comprehensively translated tissue-based microRNA (miRNA) and messenger RNA (mRNA) expression biomarkers into a liquid biopsy assay.

Methods:

We analyzed 186 clinical ESCC samples, which included 128 formalin-fixed paraffin-embedded and a matched subset of 58 serum samples, from 2 independent institutions. We performed quantitative reverse-transcription polymerase chain reaction, and developed a resistance-prediction model using the logistic regression analyses.

Results:

We first evaluated the potential of 4-miRNAs and 3-mRNAs panel, which robustly predicted resistance to NAT [area under the curve (AUC): 0.85]. Moreover, addition of tumor size to this panel increased predictive potential to establish a combination signature (AUC: 0.92). We successfully validated this signature performance in independent cohort, and our model was more accurate when the signature was combined with clinical predictors (AUC: 0.81) to establish a NAT resistance risk (NATRR) model. Finally, we successfully translated our NATRR model into a liquid biopsy assay (AUC: 0.78), and a multivariate regression analysis revealed this model as an independent predictor for response to NAT (odds ratio: 6.10; P < 0.01).

Conclusions:

We successfully developed a liquid biopsy-based assay that allows robust prediction of response to NAT in ESCC patients, and our assay provides fundamentals of developing precision-medicine.
Design and Implementation of a Career Development Program for Physician-Scientists: Lessons Learned
Authors: Kameny Rebecca R. , Amundsen Cindy L.
Published: June 2022

Importance

Although skills in health services research and data science have great potential to advance the field of urogynecology, few clinical researchers obtain such training.

Objectives

The aim of the R25 UrogynCREST Program is to prepare the next generation of physician-scientists for a successful career in urogynecologic health services research through skilled mentoring and advanced training. The purpose of this report is to describe program implementation and lessons learned.

Study Design

Administered through the program institution and in partnership with the American Urogynecologic Society, this program provided junior faculty with advanced online training and, through a core facility, access to health care databases for research projects. Participants received individualized mentoring and biostatistical support. Anonymous surveys captured actionable, real-time feedback from participants as they moved through the program.

Results

Despite a limited budget, UrogynCREST maintained a core of excellent faculty, high-quality biostatistical support, and engaged, knowledgeable advisors and mentors. This allowed for similar experiences across cohorts while permitting program improvements between cohorts in faculty-participant interactions, team dynamics, and data and regulatory support. Administrative management by a single institution facilitated responses to fiscal and regulatory changes. Asynchronized learning and partnering with a society attracted a diverse group of physician-scientists.

Conclusions

Career development programs that incorporate online education, mentoring, database access, and biostatistical support must be prepared for midprogram changes. Regular communication among stakeholders was vital. Working with a core facility provided efficient database access, but evolving regulatory and administrative processes and costs presented challenges. Our experiences implementing this program can benefit similar programs that train early-career physician-scientists.
Patient Satisfaction, Provider Productivity, and Efficacy of a Standardized Preparatory Toolkit With Broad Telemedicine Adaptation in an Outpatient Urology Clinic
Authors: Overholt Tyler L. , Dutta Rahul , Smith Whitney R. , Velet Liliya P. , Xu Raymond , Matthews Catherine A.
Published: June 2022

Importance

The COVID-19 pandemic prompted telemedicine adoption. In March 2020, we developed an implementation toolkit with a nursing protocol for patient preparation before tertiary care urology clinic visits.

Objectives

Our primary objective was to determine patient satisfaction after implementation of a telemedicine toolkit. Our secondary objective was to assess downstream productivity generated from telehealth visits.

Study Design

We prospectively conducted a postvisit survey that included the Telehealth Usability Questionnaire, a validated survey assessing patient satisfaction, for all patients with documented completion of the nursing protocol to assess patient experience and satisfaction. We then performed a retrospective chart review of all telemedicine visits to determine downstream outcomes, including imaging and procedure scheduling.

Results

Between April and May 2020, 1,422 visits were completed, of which 265 had complete nursing protocol documentation. Eighteen of 265 (6.8%) reported setup assistance. Four (1.8%) were unsuccessful and converted to a nonvisual phone visit. Overall, 186 (70.1%) completed the Telehealth Usability Questionnaire with a mean score of 118.31 ± 23.44. High satisfaction was reported regardless of race, marital status, income, education, employment status, or travel distance. Younger age (P = 0.017) and female sex (P = 0.017) were associated with greater satisfaction. Of 1,422 total visits, imaging was ordered in 29%, office procedures scheduled in 14%, and surgery scheduled in 14%. New visits were more likely to result in procedure and surgery scheduling than returns (P < 0.0001).

Conclusions

Our telemedicine toolkit designed to maximize patient engagement was successful in achieving patient-provider connectivity in 98% of patients with high satisfaction. Telemedicine visits are effective to provide comprehensive urologic care with implications beyond the pandemic.
Linguistic Differences by Gender in Letters of Recommendation for Female Pelvic Medicine and Reconstructive Surgery Fellowship Applicants From 2010 to 2020
Authors: Tappy Erryn , Pan Evelyn , Brown Larry Steven , Wang Angela , Verma Diksha , Florian-Rodriguez Maria
Published: June 2022

Importance

Linguistic differences suggestive of gender bias have been detected in letters of recommendation (LOR) for female and male residency and fellowship program applicants within multiple medical specialties.

Objective

The aim of the study was to determine whether linguistic differences exist in LOR for female and male physicians applying to female pelvic medicine and reconstructive surgery (FPMRS) fellowship.

Study Design

A retrospective analysis of FPMRS fellowship applications submitted to a university-affiliated academic center from 2010 to 2020 was performed. Linguistic Inquiry and Word Count, a text analysis software, was used to characterize the linguistic content of letters. Multivariable analysis was used to compare letter characteristics with applicant and letter writer demographics.

Results

Of 306 applications reviewed, 221 (72.2%) applicants were female and 85 (27.8%) were male. Of the 1,062 letters analyzed, 457 (43.0%) were written by female letter writers, 586 (55.2%) by males, and 19 (1.8%) were a combination. Multivariable analysis controlling for race, Step 1 score, and letter writer gender demonstrated more frequent use of affiliation words for female applicants compared with males (3.1% ± 0.3 vs. 2.9% ± 0.3, P = 0.02). No additional differences were noted in average letter length or all other linguistic categories analyzed. Multiple differences were detected between writing styles of female and male letter writers, including average letter length, use of multiple word categories, and use of communal (relationship-oriented) language. Data were stratified into 2-year periods and no longitudinal trends in linguistic differences were detected.

Conclusions

No linguistic differences, suggestive of gender bias, were found between female and male applicants to FPMRS fellowship.
Patients With Better Outcome Have Higher ERP Response to Emotional Auditory Stimuli
Authors: Portnova Galina V. , Podlepich Vitaliy V. , Skorokhodov Ivan V.
Published: June 2022

Purpose:

Accuracy of outcome prognosis is one of the most important tasks of coma arousal therapy. Reactions toward sensory stimuli are the most significant predictor of conscience and cognitive functions restoration after a brain injury. A paradigm that includes ERP registration has the advantage of detailed stimuli processing visualization. The authors aimed to investigate perception and distinguishing of emotionally significant sounds (crying and laughter) in coma patients with different consciousness restoration prognosis.

Methods:

EEG was recorded in 24 comatose patients with different outcomes (scored with Glasgow Outcome Scale-Extended) and 32 healthy volunteers. The authors presented sounds of crying and laughter. ERPs for sound stimulation were calculated.

Results:

An analysis of the correlation of ERP components and Glasgow Outcome Scale-Extended score was performed. P200 (r = 0.6, P = 0.0014) and N200 amplitudes (r = −0.56, P = 0.0037) for emotional sounds correlated with the Glasgow Outcome Scale-Extended score. The significant differences of P300 and N400 amplitudes corresponded to differences of response between sounds of crying and laughter in subjects of the control group. Unlike the control group, comatose participants with good outcome produced similar electrical activity toward pleasant and unpleasant emotional stimuli.

Conclusions:

Comatose patients with good outcome produced more prominent ERP for emotional sounds. Even the good outcome participants were unable to distinguish emotional sounds of different moods, which indicate the preservation of solely robust mechanisms of sound processing. N200 and P200 amplitudes for emotional stimuli correlated significantly with outcome prognosis in coma patients.
Identifying Suspected Volume Conduction Contamination of External Anal Sphincter Motor Evoked Potentials in Lumbosacral Spine Surgery
Authors: Wilkinson Marshall , Ajoku Uchenna , Pederson Kristine , McIntrye Ian , Zarrabian Mohammad
Published: June 2022

Purpose:

Iatrogenic injury to sacral nerve roots poses significant quality of life issues for patients. Motor evoked potential (MEP) monitoring can be used for intraoperative surveillance of these important structures. We hypothesized that volume conducted depolarizations from gluteus maximus (GM) may contaminate external anal sphincter (EAS) MEP results during lumbosacral spine surgery.

Methods:

Motor evoked potential from the EAS and medial GM in 40 patients were prospectively assessed for inter-muscle volume conduction during lumbosacral spine surgeries. Peak latency matching between the EAS and GM MEP recordings conditionally identified volume conduction (VC+) or no volume conduction (VC−). Linear regression and power spectral density analysis of EAS and medial GM MEP amplitudes were performed from VC+ and VC− data pairs to confirm intermuscle electrical cross-talk.

Results:

Motor evoked potential peak latency matching identified putative VC+ in 9 of 40 patients (22.5%). Mean regression coefficients (r
2) from peak-to-peak EAS and medial GM MEP amplitude plots were 0.83 ± 0.04 for VC+ and 0.34 ± 0.06 for VC− MEP (P < 0.001). Power spectral density analysis identified the major frequency component in the MEP responses. The mean frequency difference between VC+ EAS and medial GM MEP responses were 0.4 ± 0.2 Hz compared with 3.5 ± 0.6 Hz for VC− MEP (P < 0.001).

Conclusions:

Our data support using peak latency matching between EAS and GM MEP to identify spurious MEP results because of intermuscle volume conduction. Neuromonitorists should be aware of this possible cross-muscle conflict to avoid interpretation errors during lumbosacral procedures using EAS MEP.
Critically Appraising the Quality of Reporting of American College of Surgeons TQIP Studies in the Era of Large Data Research
Authors: Gebran Anthony , Bejjani Antoine , Badin Daniel , Sabbagh Hadi , Mahmoud Tala , El Moheb Mohamad , Nederpelt Charlie J , Joseph Bellal , Nathens Avery , Kaafarani Haytham MA
Published: June 2022

BACKGROUND:

The American College of Surgeons-Trauma Quality Improvement Program (ACS-TQIP) database is one of the most widely used databases for trauma research. We aimed to critically appraise the quality of the methodological reporting of ACS-TQIP studies.

STUDY DESIGN:

The ACS-TQIP bibliography was queried for all studies published between January 2018 and January 2021. The quality of data reporting was assessed using the Strengthening the Reporting of Observational studies in Epidemiology-Reporting of Studies Conducted Using Observational Routinely Collected Health Data (STROBE-RECORD) statement and the JAMA Surgery checklist. Three items from each tool were not applicable and thus excluded. The quality of reporting was compared between high- and low-impact factor (IF) journals (cutoff for high IF is >90th percentile of all surgical journals).

RESULTS:

A total of 118 eligible studies were included; 12 (10%) were published in high-IF journals. The median (interquartile range) number of criteria fulfilled was 5 (4–6) for the STROBE-RECORD statement (of 10 items) and 5 (5–6) for the JAMA Surgery checklist (of 7 items). Specifically, 73% of studies did not describe the patient population selection process, 61% did not address data cleaning or the implications of missing values, and 76% did not properly state inclusion/exclusion criteria and/or outcome variables. Studies published in high-IF journals had remarkably higher quality of reporting than those in low-IF journals.

CONCLUSION:

The methodological reporting quality of ACS-TQIP studies remains suboptimal. Future efforts should focus on improving adherence to standard reporting guidelines to mitigate potential bias and improve the reproducibility of published studies.
Does Adding a Measure of Social Vulnerability to a Surgical Risk Calculator Improve Its Performance?
Authors: Dyas Adam R , Carmichael Heather , Bronsert Michael R , Henderson William G , Madsen Helen J , Colborn Kathryn L , Velopulos Catherine G , Meguid Robert A
Published: June 2022

BACKGROUND:

Emerging literature suggests that measures of social vulnerability should be incorporated into surgical risk calculators. The Social Vulnerability Index (SVI) is a measure designed by the CDC that encompasses 15 socioeconomic and demographic variables at the census tract level. We examined whether adding the SVI into a parsimonious surgical risk calculator would improve model performance.

STUDY DESIGN:

The eight-variable Surgical Risk Preoperative Assessment System (SURPAS), developed using the entire American College of Surgeons (ACS) NSQIP database, was applied to local ACS-NSQIP data from 2012 to 2018 to predict 12 postoperative outcomes. Patient addresses were geocoded and used to estimate the SVI, which was then added to the model as a ninth predictor variable. Brier scores and c-indices were compared for the models with and without the SVI.

RESULTS:

The analysis included 31,222 patients from five hospitals. Brier scores were identical for eight outcomes and improved by only one to two points in the fourth decimal place for four outcomes with addition of the SVI. Similarly, c-indices were not significantly different (p values ranged from 0.15 to 0.96). Of note, the SVI was associated with most of the eight SURPAS predictor variables, suggesting that SURPAS may already indirectly capture this important risk factor.

CONCLUSION:

The eight-variable SURPAS prediction model was not significantly improved by adding the SVI, showing that this parsimonious tool functions well without including a measure of social vulnerability.
Effect of Lactate Export Inhibition on Anaplastic Thyroid Cancer Growth and Metabolism
Authors: Zhao Bixiao , Aggarwal Abha , Im Soo-Yeon , Viswanathan Keya , Landa Iñigo , Nehs Matthew A
Published: June 2022

BACKGROUND:

Anaplastic thyroid cancer (ATC) is an aggressive malignancy without effective treatments. ATC cells demonstrate upregulated glycolysis (Warburg effect), generating lactate that is subsequently exported by monocarboxylate transporter 4 (MCT4). This study aims to determine whether MCT4 inhibition can suppress ATC growth.

STUDY DESIGN:

ATC cell lines 8505C, JL30, and TCO1 were grown in low (3 mmol/L; LG) or high (25 mmol/L; HG) glucose medium containing the lactate shuttle inhibitors acriflavine (10–25 μmol/L; ACF), syrosingopine (100 µmol/L; SYR), or AZD3965 (20 µmol/L; AZD). Lactate level and cell proliferation were measured with standard assays. Seahorse analysis was performed to determine glycolytic response.

RESULTS:

Compared with HG, addition of ACF to LG decreased lactate secretion for both 8505C (p < 10-5) and JL30 (p < 10-4) cells, whereas proliferation was also reduced (p < 10-4 and 10-5, respectively). During Seahorse analysis, addition of oligomycin increased acidification by 84 mpH/min in HG vs 10 mpH/min in LG containing ACF (p < 10-5). Treatment with LG and SYR drastically diminished 8505C and TCO1 growth vs HG (p < 0.01 for both). LG and AZD treatment also led to reduced proliferation in tested cell lines (p ≤ 0.01 for all) that was further decreased by addition of ACF (p < 10-4 vs HG, p ≤ 0.01 vs LG and AZD).

CONCLUSION:

Inhibition of lactate shuttles significantly reduced proliferation and glycolytic capacity of ATC cells in a low-glucose environment. Targeting suppression of glycolytic and lactate processing pathways may represent an effective treatment strategy for ATC.
Contemporary Matched-Cohort Comparison of Surgical Approach to Inguinal Hernia Repair: Are Minimally Invasive Approaches Associated with Higher Rates of Recurrence?
Authors: Tonelli Celsa M , Lorenzo Isabela , Bunn Corinne , Kulshrestha Sujay , Abdelsattar Zaid M , Cohn Tyler , Luchette Frederick A , Baker Marshall S
Published: June 2022

BACKGROUND:

Current studies evaluating outcomes for open, laparoscopic, and robotic inguinal hernia repair, in general, include small numbers of robotic cases and are not powered to allow a direct comparison of the 3 approaches to repair.

STUDY DESIGN:

We queried the Veterans Affairs Surgical Quality Improvement Program Database to identify patients undergoing initial elective inguinal hernia repair between 2013 and 2017. Propensity score matching and multivariable logistic regression were used to make risk-adjusted assessments of association between surgical approach and outcome.

RESULTS:

A total of 39,358 patients underwent initial elective inguinal hernia repair; 32,881 (84%) underwent an open approach, 6,135 (16%) underwent a laparoscopic approach, and 342 (1%) underwent a robotic-assisted approach. Two hundred sixty-six (1%) patients had a recurrent repair performed during follow-up. On univariate comparison, patients undergoing a robotic-assisted approach had longer operative times for unilateral repair than those undergoing either an open or laparoscopic (73 ± 31 vs 74 ± 29 vs 107 ± 41 minutes; p < 0.001) approach. On multivariable logistic regression, patients with a higher BMI had an increased adjusted risk of a postoperative complication, but there was no association between surgical approach and complication rate. Three hundred forty-two patients undergoing robotic repair were 1:3:3 propensity score matched to 1,026 patients undergoing laparoscopic and 1,026 undergoing open repair. On comparison of matched cohorts, there were no statistical differences between approaches regarding recurrence (0.6% vs 0.8% vs 0.6%, p > 0.05) or complication rate (0.6% vs 1.2% vs 1.2%, p > 0.05).

CONCLUSIONS:

In patients undergoing initial elective inguinal hernia repair, rates of hernia recurrence are low independent of surgical approach. Both robotic and laparoscopic approaches demonstrate rates of early postoperative morbidity and recurrence similar to those for the open approach. The robotic approach is associated with longer operative time than either laparoscopic or open repair.
Are Residents Able to Perform a Laparoscopic Cholecystectomy with Acceptable Outcomes When the Attending Is Not Scrubbed?
Authors: Tonelli Celsa M , Lorenzo Isabela , Bunn Corinne , Kulshrestha Sujay , Cohn Tyler , Abdelsattar Zaid , Luchette Frederick A , Baker Marshall S
Published: June 2022

BACKGROUND:

Recent socioeconomic pressures in healthcare and work hour resections have limited opportunities for resident autonomy and independent decision-making. We sought to evaluate whether contemporary senior residents are being given the opportunity to operate independently and whether patient outcomes are affected when the attending is not directly involved in an operation.

STUDY DESIGN:

The VA Surgical Quality Improvement Program (VASQIP) Database was queried to identify patients undergoing elective laparoscopic cholecystectomy between 2004 and 2019. Cases were categorized as “attending” or “resident” depending on whether the attending surgeon was scrubbed. Cohorts were 1:1 propensity score–matched (PSM) for demographics, comorbidities, and facility case-mix. Clinical outcomes for matched cohorts were compared by standard methods.

RESULTS:

There were 23,831 records for patients who underwent laparoscopic cholecystectomy; 20,568 (86%) performed with the attending scrubbed, and 3,263 (14%) without the attending scrubbed. Over time there was a significant decrease in the proportion of cases without the attending scrubbed, 18% in 2004–2009 to 13% in 2015–2019 (p < 0.001). On PSM, 3,263 patients undergoing laparoscopic cholecystectomy by the residents without the attending scrubbed were successfully matched (1:1) to cases with the attending scrubbed. On comparison of matched cohorts, procedures performed without the attending scrubbed were statistically longer (102 vs 98 minutes, p = 0.001) but with no difference in rates of postoperative complications (5% vs 5%, p = 0.9).

CONCLUSION:

In comparison with cases done with more direct attending involvement, residents perform laparoscopic cholecystectomies efficiently without increased complications. Over time, attendings are more frequently scrubbed for the operation.
Giant gar from directly above the Cretaceous–Palaeogene boundary suggests healthy freshwater ecosystems existed within thousands of years of the asteroid impact
Authors: Brownstein Chase Doran , Lyson Tyler R.
Published: June 2022

The Cretaceous–Palaeogene (K–Pg) mass extinction was responsible for the destruction of global ecosystems and loss of approximately three-quarters of species diversity 66 million years ago. Large-bodied land vertebrates suffered high extinction rates, whereas small-bodied vertebrates living in freshwater ecosystems were buffered from the worst effects. Here, we report a new species of large-bodied (1.4–1.5 m) gar based on a complete skeleton from the Williston Basin of North America. The new species was recovered 18 cm above the K–Pg boundary, making it one of the oldest articulated vertebrate fossils from the Cenozoic. The presence of this freshwater macropredator approximately 1.5–2.5 thousand years after the asteroid impact suggests the rapid recovery and reassembly of North American freshwater food webs and ecosystems after the mass extinction.
Quantity over quality? Prey-field characteristics influence the foraging decisions of little penguins (
Eudyptula minor
)
Authors: Sutton G. J. , Arnould J. P. Y.
Published: June 2022

Quantifying prey characteristics is important for understanding the foraging behaviour of predators, which ultimately influence the structure and function of entire ecosystems. However, information available on prey is often at magnitudes which cannot be used to infer the fine-scale behaviour of predators, especially so in marine environments where direct observation of predator–prey interactions is rarely possible. In the present study, animal-borne video data loggers were used to determine the influence of prey type and patch density on the foraging behaviour of the little penguin (
Eudyptula minor
), an important predator in southeastern Australia. We found that numerical density positively influenced time spent foraging at a patch. However, when accounting for calorific value in density estimates, individuals spent longer at dense patches of low-quality prey. This may reflect a trade-off between capture effort and calorific gain as lower quality prey were captured at higher rates. During the breeding season, foraging trip distance and duration is constrained by the need to return to the colony each day to feed offspring. The results of the study suggest that, under these spatio-temporal constraints, little penguins maximize foraging performance by concentrating efforts at larger quantities of prey, irrespective of their calorific quality.
Wing-feather loss in white-feathered laying hens decreases pectoralis thickness but does not increase risk of keel bone fracture
Authors: Garant Renée , Tobalske Bret W. , Sassi Neila Ben , van Staaveren Nienke , Widowski Tina , Powers Donald R. , Harlander-Matauschek Alexandra
Published: June 2022

Feather loss in domestic chickens can occur due to wear and tear, disease or bird-to-bird pecking. Flight feather loss may decrease wing use, cause pectoral muscle loss and adversely impact the keel bone to which these muscles anchor. Feather loss and muscle weakness are hypothesized risk factors for keel bone fractures that are reported in up to 98% of chickens. We used ultrasound to measure changes in pectoral muscle thickness and X-rays to assess keel bone fracture prevalence following symmetric clipping of primary and secondary feathers in white- and brown-feathered birds. Four and six weeks after flight feather clipping, pectoralis thickness decreased by approximately 5%, while lower leg thickness increased by approximately 5% in white-feathered birds. This pectoralis thickness decrease may reflect wing disuse followed by muscle atrophy, while the increased leg thickness may reflect increased bipedal locomotion. The lack of effect on muscle thickness in brown-feathered hens was probably due to their decreased tendency for aerial locomotion. Finally, pectoralis thickness was not associated with keel bone fractures in either white- or brown-feathered birds. This suggests that the white-feathered strain was more sensitive to feather loss. Future prevention strategies should focus on birds most susceptible to muscle loss associated with flight feather damage.
Of fruits and fats: high-sugar diets restore fatty acid profiles in the white adipose tissue of captive dwarf lemurs
Authors: Blanco M. B. , Greene L. K. , Ellsaesser L. N. , Schopler B. , Davison M. , Ostrowski C. , Klopfer P. H. , Fietz J. , Ehmke E. E.
Published: June 2022

Fat-storing hibernators rely on fatty acids from white adipose tissue (WAT) as an energy source to sustain hibernation. Whereas arctic and temperate hibernators preferentially recruit dietary polyunsaturated fatty acids (PUFAs), tropical hibernators can rely on monounsaturated fatty acids that produce fewer lipid peroxides during oxidation. Nevertheless, compositional data on WAT from tropical hibernators are scant and questions remain regarding fat recruitment and metabolism under different environmental conditions. We analyse fatty acid profiles from the WAT of captive dwarf lemurs (
Cheirogaleus medius
) subjected to high-sugar or high-fat diets during fattening and cold or warm conditions during hibernation. Dwarf lemurs fed high-sugar (compared to high-fat) diets displayed WAT profiles more comparable to wild lemurs that fatten on fruits and better depleted their fat reserves during hibernation. One PUFA, linoleic acid, remained elevated before hibernation, potentially lingering from the diets provisioned prior to fattening. That dwarf lemurs preferentially recruit the PUFA linoleic acid from diets that are naturally low in availability could explain the discrepancy between captive and wild lemurs' WAT. While demonstrating that minor dietary changes can produce major changes in seasonal fat deposition and depletion, our results highlight the complex role for PUFA metabolism in the ecology of tropical hibernators.
DNA damage-induced block of sporulation in Streptomyces venezuelae involves downregulation of ssgB
Authors: Falguera Jan V. T. , Stratton Kathryn J. , Bush Matthew J. , Jani Charul , Findlay Kim C. , Schlimpert Susan , Nodwell Justin R.
Published: June 2022

DNA damage often causes an arrest of the cell cycle that provides time for genome integrity to be restored. In bacteria, the classical SOS DNA damage response leads to an inhibition of cell division resulting in temporarily filamentous growth. This raises the question as to whether such a response mechanism might similarly function in naturally filamentous bacteria such as Streptomyces. Streptomyces exhibit two functionally distinct forms of cell division: cross-wall formation in vegetative hyphae and sporulation septation in aerial hyphae. Here, we show that the genotoxic agent mitomycin C confers a block in sporulation septation in

Streptomyces venezuelae

in a mechanism that involves, at least in part, the downregulation of ssgB. Notably, this DNA damage response does not appear to block cross-wall formation and may be independent of canonical SOS and developmental regulators. We also show that the mitomycin C-induced block in sporulation can be partially bypassed by the constitutive expression of ssgB, though this appears to be largely limited to mitomycin C treatment and the resultant spore-like cells have reduced viability.
Emergency department care transition barriers: a qualitative study of care partners of older adults with cognitive impairment
Authors: Gettel Cameron J. , Serina Peter T. , Uzamere Ivie , Hernandez-Bigos Kizzy , Venkatesh Arjun K. , Cohen Andrew B. , Monin Joan K. , Feder Shelli L. , Fried Terri R. , Hwang Ula
Published: June 2022

INTRODUCTION

After emergency department (ED) discharge, persons living with cognitive impairment (PLWCI) and their care partners are particularly at risk for adverse outcomes. We sought to identify the barriers experienced by care partners of PLWCI during ED discharge care transitions.

METHODS

We conducted a qualitative study of 25 care partners of PLWCI discharged from four EDs. We used the validated 4AT and care partner-completed AD8 screening tools, respectively, to exclude care partners of older adults with concern for delirium and include care partners of older adults with cognitive impairment. We conducted recorded, semi-structured interviews using a standardized guide, and two team members coded and analyzed all professional transcriptions to identify emerging themes and representative quotations.

RESULTS

Care partners’ mean age was 56.7 years, 80% were female, and 24% identified as African American. We identified four major barriers regarding ED discharge care transitions among care partners of PLWCI: 1) unique care considerations while in the ED setting impact the perceived success of the care transition, 2) poor communication and lack of care partner engagement was a commonplace during the ED discharge process, 3) care partners experienced challenges and additional responsibilities when aiding during acute illness and recovery phases, and 4) navigating the health care system after an ED encounter was perceived as difficult by care partners.

DISCUSSION

Our findings demonstrate critical barriers faced during ED discharge care transitions among care partners of PLWCI. Findings from this work may inform the development of novel care partner-reported outcome measures as well as ED discharge care transition interventions targeting care partners.
Assessment of urinary exosome NHE3 as a biomarker of acute kidney injury
Authors: Yu Yanting , Ren Zhiyun , Xie Anni , Jia Yutao , Xue Ying , Wang Ping , Ji Daxi , Wang Xiaoyan
Published: June 2022

The diagnosis of acute kidney injury (AKI) traditionally depends on the serum creatinine (SCr) and urine output, which lack sufficient sensitivity and specificity. Urinary exosome as a biomarker has unique advantages. To assessed whether urinary exosome Na+/H+ exchanger isoform 3 (NHE3) protein as a nonivasive diagnostic biomarker of AKI, we constructed 4 AKI rat models: cisplatin (7.5mg/kg) injection intraperitoneally (IP), furosemide (20mg/kg, IP) with low NaCl (0.03%) diet, low NaCl (0.03%) diet with candesartan (1mg/kg, IP) and bilaterally ischemia and reperfusion(I/R) injury for 40 minutes. Besides, we assessed 6 sepsis associated AKI patients and 6 healthy volunteers. Urinary exosomes were extracted by ultra-centrifugation and NHE3 protein abundance was tested by immunoblotting in all AKI rats and human subjects. The isolated cup-shape particles with an average diameter of 70nm and enrichment in CD63 were identified as exosomes. NHE3 abundance was 6 times higher in exosome than in original urine. In cisplatin induced AKI rats, urinary exosome NHE3 was increased at day 2, 1 day earlier than the increases of serum creatinine creatinine (SCr) and blood urea nitrogen (BUN). In additional rats, urinary exosome NHE3 decreased along with the decline of SCr after EPO pretreatment. In volume depletion AKI induced by furosemide injection with low NaCl diet, urinary exosome NHE3 expression was higher than control. In low NaCl diet with candesartan related AKI, urinary exosome NHE3 was elevated at day 5, 2 days earlier than SCr. In I/R injury AKI, urinary exosome NHE3 was also raised compared with control. In humans, the urinary exosome NHE3 level was also elevated in sepsis associated AKI patients in comparison with the healthy volunteers. Then urinary exosome NHE3 protein may be used as a noninvasive diagnostic biomarker of AKI.

Impact Statement

The high non-recognition acute kidney injury (AKI) is due to lacking of alarming symptoms or specific early biomarkers. Urinary exosome as a biomarker has unique advantages. In our study, we detected urinary exosome NHE3 protein abundance in 4 different cause of AKI rat model. Urinary exosome NHE3 was increased in all 4 AKI, and even elevated earlier than SCr in some cases. Another novel point was we established a new AKI model of low NaCl diet with candesartan injection, which was common in patients with hypertension or proteinuria clinically. The detailed methods and mechanisms of this new AKI model were presented in another article being submitted. Third, we are not limited to animals, but also selected sepsis associated AKI patients to study. The conclusion that urinary exosome NHE3 may be used as a diagnosis biomarker of AKI has important clinical application value.
Acute sleep deprivation induces synaptic remodeling at the soleus muscle neuromuscular junction in rats
Authors: Sharma Binney , Roy Avishek , Sengupta Trina , Vishwakarma Lal Chandra , Singh Anuraag , Netam Ritesh , Nag Tapas Chandra , Akhtar Nasreen , Mallick Hruda Nanda
Published: June 2022

Sleep is important for cognitive and physical performance. Sleep deprivation not only affects neural functions but also results in muscular fatigue. A good night’s sleep reverses these functional derangements caused by sleep deprivation. The role of sleep in brain function has been extensively studied. However, its role in neuromuscular junction or skeletal muscle morphology is sparsely addressed although skeletal muscle atonia and suspended thermoregulation during rapid eye movement sleep possibly provides a conducive environment for the muscle to rest and repair; somewhat similar to slow-wave sleep for synaptic downscaling. In the present study, we have investigated the effect of 24 h sleep deprivation on the neuromuscular junction morphology and neurochemistry using electron microscopy and immunohistochemistry in the rat soleus muscle. Acute sleep deprivation altered synaptic ultra-structure viz. mitochondria, synaptic vesicle, synaptic proteins, basal lamina, and junctional folds needed for neuromuscular transmission. Further acute sleep deprivation showed the depletion of the neurotransmitter acetylcholine and the overactivity of its degrading enzyme acetylcholine esterase at the neuromuscular junction. The impact of sleep deprivation on synaptic homeostasis in the brain has been extensively reported recently. The present evidence from our studies shows new information on the role of sleep on neuromuscular junction homeostasis and its functioning.

Statement of significance

Sleep causes synaptic downscaling in the brain, and allows the brain to carry out various housekeeping functions. Here we have reported that the function of the sleep-wake cycle on the synaptic homeostasis extends beyond the brain. Acute sleep deprivation caused significant alteration at ultra and macrostructure of antigravity muscle and the neuromuscular junction along with adaptation to new fiber type in rats. These morpho-functional changes were well correlated with the biochemical assessment of the acetylcholine at the neuromuscular junction. These changes were partially recovered when the rats were allowed to recover from sleep deprivation. The findings suggest a new avenue for a sleep study; employing the neuromuscular junction for exploring the effect of sleep at energy and synaptic homeostasis levels.
Allele-specific gene regulation, phenotypes, and therapeutic vulnerabilities in estrogen receptor alpha mutant endometrial cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E. K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: June 2022

Activating estrogen receptor alpha (ER) mutations are present in primary endometrial and metastatic breast cancers, promoting estrogen-independent activation of the receptor. Functional characterizations in breast cancer have established unique molecular and phenotypic consequences of the receptor, yet the impact of ER mutations in endometrial cancer has not been fully explored. In this study, we used CRISPR-Cas9 to model the clinically prevalent ER-Y537S mutation and compared results to ER-D538G to discover allele-specific differences between ER mutations in endometrial cancer. We found that constitutive activity of mutant ER resulted in changes in the expression of thousands of genes, stemming from combined alterations to ER binding and chromatin accessibility. The unique gene expression programs resulted in ER mutant cells developing increased cancer associated phenotypes, including migration, invasion, anchorage independent growth, and growthin vivo. To uncover potential treatment strategies, we identified ER associated proteins via Rapid Immunoprecipitation and Mass Spectrometry of Endogenous Proteins (RIME) and interrogated two candidates, CDK9 and NCOA3. Inhibition of these regulatory proteins resulted in decreased growth and migration, representing potential novel treatment strategies for ER mutant endometrial cancer.

Implications

This study provides insight into mutant ER activity in endometrial cancer and identifies potential therapies for women with ER mutant endometrial cancer.

STATEMENT OF SIGNIFICANCE

Activating estrogen receptor alpha (ER) mutations promote ligand-independent activity of the receptor. This study evaluates ER-Y537S and ER-D538G mutations in primary endometrial cancer, revealing their effects on gene regulation and cancer-associated phenotypes. By identifying ER associated proteins, we also uncover potential novel treatments for women with ER mutant endometrial cancer.
Assembly of multi-subunit fusion proteins into the RNA-targeting type III-D CRISPR-Cas effector complex
Authors: Schwartz Evan A. , Bravo Jack P.K. , Macias Luis A. , McCafferty Caitlyn L. , Dangerfield Tyler L. , Walker Jada N. , Brodbelt Jennifer S. , Fineran Peter C. , Fagerlund Robert D. , Taylor David W.
Published: June 2022

CRISPR (Clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems are a type of adaptive immune response in bacteria and archaea that utilize crRNA (CRISPR RNA)-guided effector complexes to target complementary RNA or DNA for destruction. The prototypical type III-A and III-B CRISPR-Cas systems utilize multi-subunit effector complexes composed of individual proteins to cleave ssRNA targets at 6-nt intervals, as well as non-specifically degrading ssDNA and activating cyclic oligoadenylate (cOA) synthesis. Recent studies have shown that type III systems can contain subunit fusions yet maintain canonical type III RNA-targeting capabilities. To understand how a multi-subunit fusion effector functions, we determine structures of a variant type III-D effector and biochemically characterize how it cleaves RNA targets. These findings provide insights into how multi-subunit fusion proteins are tethered together and assemble into an active and programmable RNA endonuclease, how the effector utilizes a novel mechanism for target RNA seeding, and the structural basis for the evolution of type III effector complexes. Furthermore, our results provide a blueprint for fusing subunits in class 1 effectors for design of user-defined effector complexes with disparate activities.

Important note

While this manuscript was in preparation, a manuscript describing the structure of the type III-E effector was published1. We reference these important findings; however, a careful comparison of the structures will follow once the coordinates have been released by the PDB.
Basis for antibody- and hormone-mediated activation of TSHR in Graves’ disease
Authors: Duan Jia , Xu Peiyu , Luan Xiaodong , Ji Yujie , Yuan Qingning , He Xinheng , Jin Ye , Cheng Xi , Jiang Hualiang , Zhang Shuyang , Jiang Yi , Xu H. Eric
Published: June 2022

Thyroid stimulating hormone (TSH), through activation of its G protein-coupled receptor TSHR, controls the synthesis of thyroid hormone (TH), an essential metabolic hormone. Aberrant signaling of TSHR by autoantibodies causes Graves’ disease and hypothyroidism that affect millions of patients worldwide. Here we report the active structures of TSHR with TSH and an activating autoantibody M22, both bound to an allosteric agonist ML-109, as well as an inactive TSHR structure with inhibitory antibody K1-70. Both TSH and M22 push the extracellular domain (ECD) of TSHR into the upright active conformation. In contrast, K1-70 blocks TSH binding and is incapable of pushing the ECD to the upright conformation. Comparisons of the active and inactive structures of TSHR with those of the luteinizing hormone–choriogonadotropin receptor (LHCGR) reveal a universal activation mechanism of glycoprotein hormone receptors, in which a conserved 10-residue fragment (P10) from the hinge C-terminal loop mediated interactions from the receptor ECD to its transmembrane domain. One surprisingly feature is that there are over 15 cholesterols surrounding TSHR, supporting its preferential location in lipid rafts. These structures also highlight a common mechanism for TSH and autoantibody M22 to activate TSHR, thus providing the molecular basis for Graves’ disease.
Structural analysis of SALL4 zinc-finger domain reveals a link between AT-rich DNA binding and Okihiro syndrome
Authors: Watson James A. , Pantier Raphaël , Jayachandran Uma , Chhatbar Kashyap , Alexander-Howden Beatrice , Kruusvee Valdeko , Prendecki Michal , Bird Adrian , Cook Atlanta G.
Published: June 2022

Spalt-like 4 (SALL4) maintains vertebrate embryonic stem cell identity and is required for the development of multiple organs, including limbs. Mutations in SALL4 are associated with Okihiro syndrome and SALL4 is also a known target of thalidomide. SALL4 protein has a distinct preference for AT-rich sequences, recognised by a pair of zinc fingers at the C-terminus. However, unlike many characterised zinc finger proteins, SALL4 shows flexible recognition with many different combinations of AT-rich sequences being targeted. SALL4 interacts with the NuRD corepressor complex which potentially mediates repression of AT-rich genes. We present a crystal structure of SALL4 C-terminal zinc fingers with an AT-rich DNA sequence, which shows that SALL4 uses small hydrophobic and polar side chains to provide flexible recognition in the major groove. Missense mutations reported in patients that lie within the C-terminal zinc fingers reduced overall binding to DNA but not the preference for AT-rich sequences. Furthermore, these mutations altered association of SALL4 with AT-rich genomic sites, providing evidence that these mutations are likely pathogenic.