bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of Optic Disc Edema in Long-Duration Spaceflight Crewmembers Using Retinal Photography
Authors: Valencia William E. , Mason Sara S. , Brunstetter Tyson J. , Sargsyan Ashot E. , Schaefer Caroline M. , Tarver William J. , Van Baalen Mary G. , Gibson Charles R. , Lee Andrew G. , Danilichev Sergey N. , Hinton Patricia V. , Makarov Igor A. , Matveev Vladimir P. , Stern Claudia H. , Taniguchi-Shinojima Ari , Feldon Steven E.
Published: February 2023

Background:

Long-duration spaceflight crewmembers are at risk for spaceflight-associated neuro-ocular syndrome (SANS). One of the earliest manifestations of SANS is optic disc edema (ODE), which could be missed using the subjective Frisén scale. The primary objective of this study is to determine the inter-rater and intrarater reliability of Frisén grade for SANS-induced ODE among a trained observer cohort. The secondary objective is to propose a standardized evaluation process for SANS-induced ODE across International Space Station Partner Agencies.

Methods:

Retrospective, double-blinded diagnostic study. Preflight and postflight fundus photographs were presented to subject matter experts who identified and graded ODE. Pairs of images were also compared side-by-side for disc ranking. Grader concordance was assessed for Frisén grading and disc ranking.

Results:

Expert graders identified Grade 1 ODE in 17.35% of images from 62 crewmembers (9 female, mean [SD] age, 47.81 [5.19] years). Grades 2 and 3 were identified less than 2% of the time. Concordance in Frisén grades among pairs of graders was 70.99%. Graders identified a difference in preflight and postflight fundus photographs 17.21% of the time when using disc ranking. Pairs of graders had complete concordance in disc ranking 79.79% of the time. Perfect intrarater agreement between Frisén grade and disc ranking occurred 77.7% of the time.

Conclusions:

These findings demonstrate intergrader and intragrader variability when using the Frisén scale to identify SANS-induced ODE, which is typically milder in presentation than terrestrial cases of idiopathic intracranial hypertension. It is possible to miss early ODE on fundoscopy alone, making it insufficient as a sole criterion for the diagnosis of SANS. A more sensitive and objective method of surveillance is necessary to monitor international crewmembers for ODE, perhaps using a multimodal approach that includes technology such as optical coherence tomography.
Intracranial Pressure Fluctuations Mimicking Seizures in a Patient With Shunt Malfunction
Authors: Abramovici Sergiu , Blackburn Kaitlyn , Phinney Trevor , Mayoglou Lazarus
Published: February 2023

This case describes a patient with new onset of multiple daily paroxysmal stereotyped episodes with alteration of arousal and responsiveness. These episodes pose a diagnostic challenge because they may be misdiagnosed as epileptic seizures because of stereotypy, similarities in semiology, risk factors such as structural abnormality, and rhythmic EEG pattern. A 20-year-old woman with Chiari malformation, spina bifida, and ventriculoperitoneal shunt presented with paroxysmal episodes of change in responsiveness, concerning for seizure activity. Anti-seizure medication was started without amelioration. During the stereotyped episodes, she suffered from paroxysmal headache, drowsiness, poor responsiveness, and relative bradycardia. EEG confirmed relative bradycardia and revealed rapid buildup of generalized rhythmic delta activity without ictal features. Improvement with ICP lowering, and lack of epileptiform activity on EEG or localizing ictal semiology, prompted obtaining shunt imaging and brain MRI. The patient was subsequently diagnosed with shunt malfunction and underwent shunt revision, resulting in complete resolution of her paroxysmal spells. Although common differential diagnosis of brief paroxysmal stereotyped spells includes seizures or psychogenic nonepileptic attacks, this patient's case demonstrates clinical paroxysms caused by intracranial pressure fluctuations (plateau waves). When evaluating patients with possible intracranial pressure abnormality for paroxysmal spells, shunt malfunction should be considered as well.
Silent information regulator 1 mediates H2S-inhibited chronic restraint stress-induced depressive-like behaviors by regulating hippocampal autophagy
Authors: Du Lv , Chen Lei , Luo Bang , Chen Yong-Jun , Zhang Ping , Tang Yi-Yun , Jiang Jia-Mei , Zou Wei
Published: February 2023

Objectives

Our previous study has demonstrated that hydrogen sulfide (H2S), a novel gasotransmitter, attenuates excessive autophagy and depressive-like behaviors in chronic restraint stress (CRS)-exposed rats, but the underlying molecular mechanism remains to be elucidated. Silent information regulator 1 (SIRT1), a deacetylase at the consumption of NAD+ plays an important regulatory role in depression. Hence, this study aimed to investigate whether SIRT1 mediates the protective effect of H2S on CRS-induced depressive-like behaviors by regulating hippocampal autophagy.

Methods

Adult male Sprague-Dawley (SD) rats were subjected to CRS (6 h × 28 days) to induce depression-like behavior. Rats were injected with sodium hydrosulfate (NaHS, 100 μmol/kg/d, i.p.), as a donor of H2S, alone or in combination with Sirtinol (a SIRT1 inhibitor; 10 nmol, i.c.v.) during CRS process. The depression-like characteristics of rats were assessed by the novelty-suppressed feeding test (NSFT), tail suspension test (TST), forced swimming test (FST) and open field test (OFT). The number of hippocampal autophagosomes was detected by transmission electron microscopy. The expressions of hippocampal autophagy-related proteins were measured by western blotting analysis.

Results

Sirtinol blocked the inhibitory effect of H2S on depressive-like behaviors in CRS-exposed rats according to NSFT, TST, FST and OFT. In addition, sirtinol reversed the protective response of H2S to CRS-induced excessive autophagy, as proved by the increases in the number of autophagosomes and the expression of Beclin-1 as well as a decrease in the expression of P62 in the hippocampus.

Conclusion

These results indicated that SIRT1 contributes to the antidepressant-like function of H2S during CRS via reducing hippocampal autophagy.
Symptom Burden in Patients with Malignant Bowel Obstruction Treated With or Without Surgery
Authors: Ayabe Reed , Mendoza Tito , Yennu Sriram , Bruera Eduardo , Williams Loretta A , Badgwell Brian
Published: February 2023

Background:

Gastrointestinal obstruction is the most common indication for palliative surgical consultation. We sought to assess patient-reported outcomes (PRO) and survival after surgical and non-surgical treatment of malignant bowel obstruction.

Study Design:

This was a prospective observational study enrolling patients with advanced malignancy who underwent surgical consultation at a tertiary cancer center. PROs were evaluated using a previously validated inventory, the MDASI-GIO, administered at enrollment and 7 other time points for up to 90 days.

Results:

We enrolled 125 patients, of whom 37 underwent surgery and 88 did not. Patients treated non-surgically were more likely to have carcinomatosis on imaging (71% vs 49%, P=0.02). Pain medicine, palliative care, and chaplaincy consultations occurred in 17%, 30%, and 15% of patients within the first month of enrollment. Higher mean symptom scores were noted by surgical patients although the only single scores with effect sizes ≥0.5 were symptom interference with general activity and work. The composite score for interference in work, activity, and walking had the largest effect size at -0.37, indicating greater interference in patients undergoing surgery. Patients selected for surgery had extended overall survival (median 15 vs 3 months, P<0.01). Carcinomatosis, palliative care evaluation, and venting gastrostomy tube were associated with increased risk of death, while ability to receive subsequent chemotherapy and surgical management were positive prognostic indicators.

Conclusion:

In this first study evaluating PROs after treatment for malignant bowel obstruction we found that selection for surgical treatment was associated with improved survival, but also more symptom interference in general activities and work. These results may be useful in palliative surgical decision-making and informing patients during consultation for malignant bowel obstruction.
Less Is More: Parathyroidectomy and the Association with Postoperative Hypocalcemia in Dialysis Patients
Authors: Wang Rongzhi , Disharoon Mitchell , Frazier Rachel , Xie Rongbing , Moses Cara , Gillis Andrea , Fazendin Jessica , Lindeman Brenessa , Gutierrez Orlando M , Chen Herbert
Published: February 2023

Background:

Parathyroidectomy (PTx) is the most effective treatment for secondary hyperparathyroidism (HPT). Literature regarding the effect of surgical approaches on postoperative hypocalcemia is limited and mainly focuses on postoperative calcium levels. This study aims to evaluate the association of subtotal PTx and total PTx with auto-transplantation (AT) for secondary HPT with postoperative hypocalcemia.

Study Design:

We reviewed all dialysis patients who underwent PTx (n=143) at our institution from 2010 to 2021. Postoperative hypocalcemia adverse events were defined as postoperative intravenous (IV) calcium requirement or 30-day readmission due to hypocalcemia. Postoperative hypocalcemia adverse events, length of stay, and oral calcium requirement at 1-month follow-up were compared between the two groups.

Results:

Of the 143 patients, 119(83.2%) underwent total PTx with AT, and 24(16.8%) underwent subtotal PTx. Patients who underwent subtotal PTx had shorter mean (±SD) length of stay (1.8 ± 1.7 vs. 3.5 ± 3.2, p=0.002), were less likely to develop hypocalcemia adverse events (8.3% vs. 47.1%, p<0.001) and required less median elemental calcium supplementation at 1-month follow-up (1558 vs.3193 mg, p<0.001). There was no significant difference in surgical success between the two groups (91.7% vs. 89.1%, p=0.706). Stepwise multivariable regression demonstrated that patients who underwent total PTx with AT were 11.9 times more likely to develop hypocalcemia adverse events (aOR 11.9 95%CI 2.2-66.2, p=0.004), had 1.24-day longer length of stay (95%CI 0.04-2.44, p=0.044) and required 1776.1 mg more elemental calcium (95%CI 661.5-2890.6 mg, p=0.002).

Conclusion:

Subtotal parathyroidectomy is associated with less postoperative hypocalcemia and provides similar surgical cure for dialysis patients with secondary hyperparathyroidism.
Thirty-Day Perioperative Clinical Outcomes of Transcarotid Artery Revascularization vs Carotid Endarterectomy in a Single-Center Experience
Authors: AbuRahma Ali F , Santini Adrian , AbuRahma Zachary T , Lee Andrew , Seal Kimberly , Veith Christina , Dean Scott , Davis Elaine
Published: February 2023

Background:

Transcarotid Artery Revascularization (TCAR) has been proposed as a alternative to carotid endarterectomy (CEA) and transfemoral carotid artery stenting in high-risk patients. Recently CMS expanded coverage for TCAR to include standard surgical risk patients within the SVS Vascular Quality Initiative TCAR surveillance project (TSP). Few single centers compared the clinical outcome of TCAR to CEA. This study compares 30-day perioperative clinical outcomes between TCAR and CEA.

Study Design:

This is retrospective analysis of prospectively collected data from TSP of TCAR patients enrolled in our institution and compared with CEAs done in the same time/providers. The primary outcome was stroke and/or death. Secondary outcome included stroke, death, MI, cranial nerve injury (CNI), bleeding, etc. Propensity matching was done to analyze outcomes.

Results:

The study analyzed 501 patients (347 CEA, 154 TCAR). There were no significant differences in symptomatic status (43% for CEA vs 38% for TCAR, p 0.3032). TCAR had higher patients with hypertension (p 0.0395), CAD (p 0.028), CHF (p 0.0392). The 30-day perioperative complication rates for CEA vs TCAR were: stroke 1% vs 3% (p 0.1416), stroke/death 1% vs 3% (p 0.1854), MI 0.6% vs 0.7% (p 1), death 0.6% vs 0% (p 1), stroke/death/MI 2% vs 4% (p 0.233), CNI 4% vs 2% (p 0.4117), major hematoma (requiring re-intervention) 2% vs 3% (p 1). After matching 154 CEA patients and 154 TCAR, 30-day perioperative complication rates were: stroke 2% vs 3% (p 0.7229), stroke/death 3% vs 3% (p 1), death 1.3% vs 0% (p 0.4984), MI 0.7% vs 0.7% (p 1), stroke/death/MI 3% vs 4% (p 0.7588).

Conclusion:

This study showed that using propensity match analysis both CEA and TCAR have similar 30-day perioperative outcomes. Further long-term data is needed.
Combined Tumor Burden Score and Carbohydrate Antigen 19-9 Grading System to Predict Outcomes Among Patients with Intrahepatic Cholangiocarcinoma
Authors: Moazzam Zorays , Alaimo Laura , Endo Yutaka , Lima Henrique A , Ruzzenente Andrea , Guglielmi Alfredo , Aldrighetti Luca , Weiss Matthew , Bauer Todd W , Alexandrescu Sorin , Poultsides George A , Maithel Shishir K , Marques Hugo P , Martel Guillaume , Pulitano Carlo , Shen Feng , Cauchy François , Koerkamp Bas Groot , Endo Itaru , Cloyd Jordan , Ejaz Aslam , Pawlik Timothy M
Published: February 2023

Background:

The interplay of CA19-9 and tumor burden score (TBS) in intrahepatic cholangiocarcinoma (ICC) remains ill-defined. We evaluated the role of TBS and CA19-9 relative to overall survival (OS) and recurrence, as well as the predictive ability of the Combination of TBS and CA 19-9 (CTC) grading system.

Study Design:

Patients who underwent liver resection for ICC between 2000-2020 were identified using a multi-institutional database. The impact of CA19-9 and TBS on 5-year OS and 3-year recurrence was assessed, along with the prognostic accuracy of the CTC system (a composite score of CA19-9 and TBS).

Results:

Among 831 patients, the median age was 58.2 years and 482 (58.0%) were male. The median CA19-9 level was 49.7 (17.0–221.0) U/mL, while median TBS was 6.1 (IQR 4.1–8.3). Median and 5-year OS were 36.9 (IQR 32.3–43.1) months and 38.9%, respectively; overall 3-year recurrence was 68.9%. Survival varied relative to CA19-9 (low: 49.0% vs. high: 19.7%) and TBS (low: 53.6% vs. high: 26.9%) had worse 5-year survival (both p<0.001). On multivariable analysis, high CA19-9 (HR 2.02, 95%CI 1.64–2.49) and high TBS (HR1.64, 95%CI 1.32–2.06) remained independently associated with OS. In turn, the CTC grading score stratified 5-year OS (low CTC: 57.7% vs. intermediate CTC: 39.9% vs. high CTC: 12.6%; p<0.001), and remained an independent prognostic factor (referent: low CTC; intermediate CTC HR 1.54, 95%CI 1.18–2.01; high CTC: HR 3.28, 95%CI 2.47–4.36).

Conclusion:

The interplay between tumor morphology and biology dictated long-term prognosis after liver resection for ICC. Prognostic models such as the CTC grading system may inform discussions around prognosis, as well as help identify which patients with ICC may benefit more from neoadjuvant chemotherapy rather than up-front surgery.
Improving Billing and Collections in a High-Volume Pediatric Surgery Practice: Denials-Based Approach
Authors: Ryan Mark L , Mutore Kevin T , DeLeon Jennifer , Gillory Lauren , Chung Dai H , Pandya Samir
Published: February 2023

Introduction:

Despite minimal coding and billing training, surgeons are frequently tasked with both in clinical practice. This often results in denials for reimbursement based upon incorrect or insufficient documentation, and reduced collections for work performed. We sought to evaluate how to correct these deficits while improving reimbursement for the most frequently rejected procedures at a high-volume academic center.

Methods:

Hospital billing data was analyzed for a four-year period (2018-2021) to determine the CPT® code denials with the largest overall cost. The denials were then stratified according to payor, reason for denial, and preventability. Assigned ICD-10 codes were categorized based on specificity as related to the procedure. The distribution of denials according to ICD-10 specificity was evaluated using the Chi-Squared test.

Results:

8,833 denials totaling $11,009,108 in billing were noted over the study period. The CPT® code 44970 (laparoscopic appendectomy) was the code associated with the largest financial impact, representing 12.8% of the total denied amount ($1.41M). Of the 823 denials for CPT® 44970, 93.3% were associated with nonspecific ICD-10 codes, whereas only 42.0% had been associated with procedure-specific ICD-10 codes. Of the patients with nonspecific codes, 80.7% of denials were due to criteria that could be remedied with supplemental information or timely filing, representing $1,059,968 in collections.

Conclusions:

This is the first study to systematically evaluate a pathway for using denial data to improve collections for work performed at a high-volume academic pediatric surgery practice. Using this methodology, targets for improvement in coding and/or documentation can be identified to improve the financial performance of a surgical department. This study also provides evidence that association with nonspecific diagnostic codes is correlated with initial denial of payment by insurance companies.
Electroconvulsive Therapy Anesthetic Choice and Clinical Outcomes
Authors: Li Kevin J. , Slama Natalie E. , Hirschtritt Matthew E. , Anshu Prachi , Iturralde Esti
Published: February 2023

Objective

Etomidate and methohexital are the 2 commonly used anesthetics for electroconvulsive therapy (ECT) in the United States. The objective of this study was to examine how anesthetic choice between etomidate and methohexital is associated with real-world clinical outcomes.

Methods

This naturalistic retrospective cohort study examined longitudinal electronic health records for 495 adult patients who received 2 or more ECT treatments from 2010 to 2019 in Kaiser Permanente North California, a large integrated health care system. Study outcomes included 12-month posttreatment depression remission as measured by the 9-item Patient Health Questionnaire, psychiatric and all-cause emergency department visits, and psychiatric and all-cause hospitalizations.

Results

Anesthetic choice was not significantly related to depression severity, emergency department visits, or psychiatric hospitalizations at 12 months after completing ECT. In exploratory analyses, we found that etomidate compared with methohexital was associated with higher rates of patient discomfort adverse effects—postictal agitation, phlebitis, and myoclonus (2.4% vs 0.4%; P < 0.001).

Conclusions

We present the first large comparison of etomidate and methohexital as anesthetics for ECT and their associations with real-world outcomes. Our study showed no significant difference on depression remission, emergency department visits, or hospitalizations 12-months posttreatment. Thus, clinicians should focus on other patient or treatment characteristics when deciding on anesthetics for ECT. Further investigation is needed to confirm our exploratory findings that etomidate use was correlated with a higher rate of patient discomfort adverse effects relative to methohexital.
Wilms tumor mutational subclasses converge to driveCCND2overexpression
Authors: Xu Lin , Desai Kavita , Kim Jiwoong , Zhou Qinbo , Guo Lei , Xiao Xue , Zhang Yanfeng , Zhou Li , Yuksel Aysen , Catchpoole Daniel R. , Amatruda James F. , Chen Kenneth S.
Published: February 2023

Wilms tumor, the most common kidney cancer in pediatrics, arises from embryonic renal progenitors. Although many patients are cured with multimodal therapy, outcomes remain poor for those with high-risk features. Recent sequencing efforts have provided few biological or clinically actionable insights. Here, we performed DNA and RNA sequencing on 94 Wilms tumors to understand how Wilms tumor mutations transform the transcriptome to arrest differentiation and drive proliferation. We show that most Wilms tumor mutations fall into four classes, each with unique transcriptional signatures: microRNA processing, MYCN activation, chromatin remodeling, and kidney development. In particular, the microRNA processing enzymeDROSHAis one of the most commonly mutated genes in Wilms tumor. We show thatDROSHAmutations impair pri-microRNA cleavage, de-repress microRNA target genes, halt differentiation, and overexpress cyclin D2 (CCND2). Several mutational classes converge to driveCCND2overexpression, which could render them susceptible to cell-cycle inhibitors.

Significance

Through integrated DNA and RNA sequencing analyses, we demonstrate novel genotype-transcriptome associations in Wilms tumors that converge to driveCCND2overexpression. Thus, although Wilms tumor mutations are not currently targetable, their effects on the transcriptome may be.
Occurrence of enteroparasites in vegetables commercialized in São Paulo, Brazil
Authors: Boccia Thais , Lopes Andrea Bento
Published: February 2023

Parasitic diseases constitute an important public health problem. Their transmission can occur by ingestion of water contaminated with human feces and/or parasitized animals, and by poorly washed fruits and vegetables. Considering the scarcity of national studies reporting the degree of contamination of vegetables, the present study investigated and compared the contamination of enteroparasites in 32 samples of each of the types: lettuce (Lactuca sativa) of the smooth and curly variety and watercress (Nasturtium officinale), commercialized in the city of São Paulo. For this purpose, the samples were washed with distilled water. The product of the washings was decanted in conical sedimentation vessels for 24 hours by the spontaneous sedimentation method, and the sediment was analyzed under an optical microscope. The results showed that of the 96 samples analyzed, 78 (82%) presented some parasitic form. Vegetables purchased from supermarkets and street markets showed a contamination rate of 80% (48/60) and 83% (30/36), respectively. The parasitic forms found were: 42%Ancylostoma splarvae,15.94% Ancylostoma speggs, 7.24%Ascaris speggs, 8.69%Enterobius speggs, 8.69 %Strongyloides splarvae, 7.24%Hymenolepis speggs, 5.79%Taenia speggs and 4.34%Entamoeba spcysts. These results demonstrate the need to develop a more efficient health education program aimed at horticulturists and vegetable handlers, with the intention of preventing the occurrence of these parasites, and further contamination of the population.
A septo-hypothalamic-medullary circuit directs stress-induced analgesia
Authors: Shah Devanshi Piyush , Sharma Pallavi Raj , Agarwal Rachit , Barik Arnab
Published: February 2023

Abstract

Stress is a potent modulator of pain. Specifically, acute stress due to physical restraint induces stress-induced analgesia (SIA). However, where and how acute stress and pain pathways interface in the brain are poorly understood. Here, we describe how the dorsal lateral septum (dLS), a forebrain limbic nucleus, facilitates SIA through its downstream targets in the lateral hypothalamic area (LHA) of mice. Taking advantage of transsynaptic viral-genetic, optogenetic, and chemogenetic techniques, we show that the dLS→LHA circuitry is sufficient to drive analgesia and is required for SIA. Further, our results reveal that the dLS→LHA pathway is opioid-dependent and modulates pain through the pro-nociceptive neurons in the rostral ventromedial medulla (RVM). Remarkably, we found that the inhibitory dLS neurons are recruited specifically when the mice struggle to escape under restraint and, in turn, inhibit excitatory LHA neurons. As a result, the RVM neurons downstream of LHA are disengaged, thus suppressing nociception. Together, we delineate a poly-synaptic pathway that can transform escape behavior in mice under restraint to acute stress into analgesia.
How the dominant reading direction changes parafoveal processing: A combined EEG/eye-tracking study
Authors: Huang Xin , Ng Hezul Tin-Yan , Lin Chien Ho , Yan Ming , Dimigen Olaf , Sommer Werner , Maurer Urs
Published: February 2023

Reading directions vary across writing systems. Through long-term experience readers adjust their visual systems to the dominant reading direction in their writing systems. However, little is known about the neural correlates underlying these adjustments because different writing systems do not just differ in reading direction, but also regarding visual and linguistic properties. Here, we took advantage that Chinese is read to different degrees in left-right or top-down directions in different regions. We investigated visual word processing in participants from Taiwan (both top-down and left-right directions) and from mainland China (only left-right direction). Combined EEG/eye tracking was used together with a saccade-contingent parafoveal preview manipulation to investigate neural correlates, while participants read 5-word lists. Fixation-related potentials (FRPs) showed a reduced late N1 effect (preview positivity), but this effect was modulated by the prior experience with a specific reading direction. Results replicate previous findings that valid previews facilitate visual word processing, as indicated by reduced FRP activation. Critically, the results indicate that this facilitation effect depends on experience with a given reading direction, suggesting a specific mechanism how cultural experience shapes the way people process visual information.
A Dual Function Antibody Conjugate Enabled Photoimmunotherapy Complements Fluorescence and Photoacoustic Imaging of Head and Neck Cancer Spheroids
Authors: Saad Mohammad A. , Grimaldo-Garcia Stacey , Sweeney Allison , Mallidi Srivalleesha , Hasan Tayyaba
Published: February 2023

Several molecular-targeted imaging and therapeutic agents are in clinical trials for image-guided surgery and photoimmunotherapy (PIT) of head and neck cancers. In this context, we have previously reported the development, characterization, and specificity of a dual function antibody conjugate (DFAC) for multi-modal imaging and photoimmunotherapy (PIT) of EGFR over-expressing cancer cells. The DFAC reported previously and used in the present study, comprises of an EGFR targeted antibody – Cetuximab conjugated to Benzoporphyrin derivative (BPD) for fluorescence imaging and PIT, and a Si-centered naphthalocyanine dye for photoacoustic imaging. We report here the evaluation and performance of DFAC in detecting microscopic cancer spheroids by fluorescence and photoacoustic imaging along with their treatment by PIT. We demonstrate that while fluorescence imaging can detect spheroids with volumes greater than 0.049 mm3, photoacoustic imaging-based detection was possible even for the smallest spheroids (0.01 mm3), developed in the study. When subjected to PIT, the spheroids showed a dose-dependent response with smaller spheroids (0.01 and 0.018 mm3) showing a complete response with no recurrence when treated with 100 J/cm2. Together our results demonstrate the complementary imaging and treatment capacity of DFAC. This potentially enables fluorescence imaging to assess tumor presence on a macroscopic scale followed by photoacoustic imaging for delineating tumor margins guiding surgical resection and elimination of any residual microscopic disease by PIT, in a single intra-operative setting.
CD39+conventional CD4+T cells with exhaustion traits and cytotoxic potential infiltrate tumors and expand upon CTLA-4 blockade
Authors: Bossio Sabrina N. , Abrate Carolina , Tosello Boari Jimena , Rodriguez Constanza , Canale Fernando P. , Ramello María C. , Richer Wilfrid , Rocha Dario , Sedlik Christine , Vincent-Salomon Anne , Borcoman Edith , Del Castillo Andres , Gruppi Adriana , Fernandez Elmer , Acosta Rodríguez Eva V. , Piaggio Eliane , Montes Carolina L.
Published: February 2023

Background

Conventional CD4+T (Tconv) lymphocytes play important roles in tumor immunity; however, their contribution to tumor elimination remains poorly understood.

Methods

Here we describe a subset of Tconv cells characterized by the expression of CD39. The phenotype, the effector function and transcriptional profile of tumor-infiltrating CD39+Tconv lymphocytes from different mouse cancer models and breast cancer patients were studied by multiparametric flow cytometry and RNA sequencing. The impact of thein vivoCTLA-4 blockade on the tumor-infiltrating CD39+Tconv population was assessed in mice grafted with the immunogenic MC38 colorectal tumor. Overall survival was evaluated in a cohort of patients from the TCGA consortium.

Results

In mouse cancer models, we observed that CD39+Tconv cells accumulated in tumors as they grew but were absent in lymphoid organs. Compared to tumor CD39−counterparts, CD39+Tconv cells exhibited a cytotoxic and exhausted signature at the transcriptomic level, confirmed by high protein expression of inhibitory receptors and transcription factors related to the exhaustion phenotype. Additionally, CD39+Tconv cells showed increased production of IFN gamma, granzyme B, perforin and CD107a expression, but reduced production of TNF.In vivoCTLA-4 blockade induced the expansion of tumor CD39+Tconv cells, which maintained their cytotoxic and exhausted features. In breast cancer patients, CD39+Tconv cells were found in tumors and in metastatic lymph nodes but were less frequent in adjacent non-tumoral mammary tissue and not detected in non-metastatic lymph nodes and blood. Human tumor CD39+Tconv cells constituted a heterogeneous cell population with features of exhaustion, impaired TNF production, and high expression of inhibitory receptors and CD107a. We found that high CD4 and ENTPD1 (CD39) gene expression in human tumor tissues correlated with a higher overall survival rate in breast cancer patients.

Conclusions

We found that CD39 acts as a biomarker of Tconv cells with characteristics of both exhaustion and cytotoxic potential. CTLA-4 blockade expands CD39+CD4+T cells which may contribute to the reduction of tumor development. Discovering the role of CD39-expressing CD4+T cells in the tumor microenvironment should help design new strategies to manipulate them and improve the efficacy of current immunotherapies.
IL-15 synergizes with CD40 agonist antibodies to induce durable immunity against bladder cancer
Authors: Wong Jeffrey L. , Smith Patrick , Angulo-Lozano Juan , Ranti Daniel , Bochner Bernard H. , Sfakianos John P. , Horowitz Amir , Ravetch Jeffrey V. , Knorr David A.
Published: February 2023

CD40 is a central co-stimulatory receptor implicated in the development of productive anti-tumor immune responses across multiple cancers, including bladder cancer. Despite strong preclinical rationale, systemic administration of therapeutic agonistic antibodies targeting the CD40 pathway have demonstrated dose limiting toxicities with minimal clinical activity to date, emphasizing an important need for optimized CD40-targeted approaches, including rational combination therapy strategies. Here, we describe an important role for the endogenous IL-15 pathway in contributing to the therapeutic activity of CD40 agonism in orthotopic bladder tumors, with upregulation of trans-presented IL-15/IL-15Rα surface complexes, particularly by cross-presenting cDC1s, and associated enrichment of activated CD8 T cells within the bladder tumor microenvironment. In bladder cancer patient samples, we identify DCs as the primary source of IL-15, however, they lack high levels of IL-15Rα at baseline. Using humanized immunocompetent orthotopic bladder tumor models, we demonstrate the ability to therapeutically augment this interaction through combined treatment with anti-CD40 agonist antibodies and exogenous IL-15, including the fully-human Fc-optimized antibody 2141-V11 currently in clinical development for the treatment of bladder cancer. Combination therapy enhances the crosstalk between Batf3-dependent cDC1s and CD8 T cells, driving robust primary anti-tumor activity and further stimulating long-term systemic anti-tumor memory responses associated with circulating memory-phenotype T and NK cell populations. Collectively, these data reveal an important role for IL-15 in mediating anti-tumor CD40 agonist responses in bladder cancer and provide key proof-of-concept for combined use of Fc-optimized anti-CD40 agonist antibodies and agents targeting the IL-15 pathway. These data support expansion of ongoing clinical studies evaluating anti-CD40 agonist antibodies and IL-15-based approaches to evaluate combinations of these promising therapeutics for the treatment of patients with bladder cancer.
Spatial uncertainty and environmental geometry in navigation
Authors: Kang Yul HR , Wolpert Daniel M , Lengyel Máté
Published: February 2023

Variations in the geometry of the environment, such as the shape and size of an enclosure, have profound effects on navigational behavior and its neural underpinning. Here, we show that these effects arise as a consequence of a single, unifying principle: to navigate efficiently, the brain must maintain and update the uncertainty about one’s location. We developed an image-computable Bayesian ideal observer model of navigation, continually combining noisy visual and self-motion inputs, and a neural encoding model optimized to represent the location uncertainty computed by the ideal observer. Through mathematical analysis and numerical simulations, we show that the ideal observer accounts for a diverse range of sometimes paradoxical distortions of human homing behavior in anisotropic and deformed environments, including ‘boundary tethering’, and its neural encoding accounts for distortions of rodent grid cell responses under identical environmental manipulations. Our results demonstrate that spatial uncertainty plays a key role in navigation.
EXPANSION OF ANTIGEN- SPECIFIC MEMORY CELLS AS A POTENTIAL BOOSTER FOR FOOD TOLERANCE INDUCTION
Authors: Pereira e Silva Airton , Marmello Bárbara Oliveira , Letícia Bentes Ana , Faccini Claudia Regina Josetti das Neves , Rodrigues Sónia Kristy Pinto Melo , Soares João Ricardo Almeida , Nicolau Campos Sylvia Maria , Teixeira Gerlinde Agate Platais Brasil
Published: February 2023

Approximately 3% of children in Western countries are diagnosed with peanut allergy, a likely lifelong disease. The preferred treatment for food allergy is allergen avoidance. However, oral immunotherapy is an FDA-approved treatment to re-induce tolerance, still, not all patients respond as expected. Thus, the aim of this work is to evaluate whether the association of an antigen-specific tolerogenic (oral tolerance) bystander effect can ameliorate the recovery of inflamed intestinal mucosa. Adult male C57BL/6 mice were divided into five groups, four of which were submitted to an intestinal inflammation induction protocol to peanuts. After sensitization, experimental groups were orally challenged with either peanuts or a hybrid diet (peanuts + mouse chow). In a second stage, groups were sensitized, challenged with peanuts, and then received either peanuts, hybrid diet, or ovalbumin chow during the recovery period of the inflamed mucosa. Results showed no changes in diet intake and body weight. Polyisotypic anti-peanut IgG and IgG1 were significantly increased in the serum from animals in allergic groups. The group that received the hybrid diet showed an increase in CD4+CD25+Foxp3+regulatory T cells, as well as in B220+CD3-CD27+memory B cells. Histology of the duodenum showed a decrease in intraepithelial leukocytes in animals who received hybrid diet. Together, our results show that when the tolerogen is added to a diet containing the allergen, it can ameliorate the induction of local inflammation. Simultaneously offering the allergen with a tolerated food increased the mucosal recovery due to the expansion of previously induced memory cells.
Kv12-Encoded K+Channels Drive the Day-Night Switch in the Repetitive Firing Rates of SCN Neurons
Authors: Hermanstyne Tracey O. , Yang Nien-Du , Granados-Fuentes Daniel , Li Xiaofan , Mellor Rebecca L. , Jegla Timothy , Herzog Erik D. , Nerbonne Jeanne M.
Published: February 2023

Considerable evidence suggests that day-night rhythms in the functional expression of subthreshold potassium (K+) channels regulate daily oscillations in the rates of spontaneous action potential firing of neurons in the suprachiasmatic nucleus (SCN), the master circadian pacemaker in mammals. The K+conductance(s) driving these daily rhythms in repetitive firing rates, however, have not been identified. To test the hypothesis that subthreshold Kv12.1/Kv12.2-encoded K+channels play a role, we obtained current-clamp recordings from SCN neurons in slices prepared from adult mice harboring targeted disruptions in theKcnh8(Kv12.1−/−) orKcnh3(Kv12.2−/−) locus. We found that mean nighttime repetitive firing rates were higher in Kv12.1−/−and Kv12.2−/−, than in wild type (WT), SCN neurons. In marked contrast, mean daytime repetitive firing rates were similar in Kv12.1−/−, Kv12.2−/−and WT SCN neurons, and the day-night difference in mean repetitive firing rates, a hallmark feature of WT SCN neurons, was eliminated in Kv12.1−/−and Kv12.2−/−SCN neurons. Similar results were obtained within vivoshRNA-mediated acute knockdown of Kv12.1 or Kv12.2 in adult SCN neurons. Voltage-clamp experiments revealed that Kv12-encoded current densities in WT SCN neurons are higher at night than during the day. In addition, pharmacological block of Kv12-encoded currents increased the mean repetitive firing rate of nighttime, but not daytime, in WT SCN neurons. Dynamic clamp-mediated subtraction of modeled Kv12-encoded currents also selectively increased the mean repetitive firing rates of nighttime WT SCN neurons. Despite the elimination of nighttime decrease in the mean repetitive firing rates of SCN neurons, however, locomotor (wheel-running) activity remained rhythmic in Kv12.1−/−, Kv12.2−/−, Kv12.1-targeted shRNA-expressing, and Kv12.2-targeted shRNA-expressing animals.
Pancreatic Deletion of Mitogen-inducible Gene 6 Promotes Beta Cell Proliferation Following Destruction
Authors: El Kimberley , Bauer Brandon M. , Chen Yi-Chun , Jeong Jae-Wook , Fueger Patrick
Published: February 2023

Type 1 Diabetes (T1D) is caused by autoimmune-mediated beta cell destruction. Following beta cell injury, the pancreas attempts to launch a cellular repair and regenerative program, yet it fails to completely restore functional beta cell mass. One component of this regenerative program is epidermal growth factor receptor (EGFR) signaling. However, upon irreparable beta cell damage, EGFR signaling is dampened, disrupting attempts to restore functional beta cell mass and maintain normoglycemia. We previously demonstrated that the negative feedback inhibitor of EGFR, Mitogen-inducible gene 6 (Mig6), is induced by the pro-inflammatory cytokines central to the autoimmune-mediated beta cell destruction. We also established that pro-inflammatory cytokines suppress EGFR activation, and siRNA-mediated suppression of Mig6 restores EGFR signaling. Thus, we hypothesized that pro-inflammatory cytokines induce nitric oxide production and that in turn induced Mig6, disrupting EGFR repair mechanisms. We determined that NO induces Mig6, attenuating EGFR signaling, and NO synthase inhibition blocks the cytokine-mediated induction of Mig6, thereby restoring cytokine-impaired EGFR signaling. To that end, we treated mice lacking pancreatic Mig6 and control mice with a streptozotocin (STZ) to induce beta cell death and diabetes in a way that mimics the onset and progression of T1D. Whereas STZ-treated control mice became hyperglycemic and had reduced beta cell mass, STZ-treated Mig6 pancreas-specific knock out (PKO) mice remained euglycemic and glucose tolerant due to preserved beta cell mass. The restoration of beta cell mass in PKO mice was accompanied by enhanced beta cell proliferation. Thus, our work suggests that Mig6 is a promising target to preserve beta cell mass before overt T1D.
Analysis of nucleotide variations in human g-quadruplex forming regions associated with disease states
Authors: Neupane Aryan , Chariker Julia H. , Rouchka Eric C.
Published: February 2023

While the role of G4 G quadruplex structures has been identified in cancers and metabolic disorders, single nucleotide variations (SNVs) and their effect on G4s in disease contexts have not been extensively studied. The COSMIC and CLINVAR databases were used to detect SNVs present in G4s to identify sequence level changes and their effect on alteration of G4 secondary structure. 37,515 G4 SNVs in the COSMIC database and 2,115 in CLINVAR were identified. Of those, 7,236 COSMIC (19.3%) and 416 (18%) of the CLINVAR variants result in G4 loss, while 2,728 (COSMIC) and 112 (CLINVAR) SNVs gain a G4 structure. The gene ontology term “GnRH (Gonadotropin-releasing hormone) secretion” is enriched in 21 genes in this pathway that have a G4 destabilizing SNV. Analysis of mutational patterns in the G4 structure show a higher selective pressure (3-fold) in the coding region on the template strand compared to the non-template strand. At the same time, an equal proportion of SNVs were observed among intronic, promoter and enhancer regions across strands. Using GO and pathway enrichment, genes with SNVs for G4 forming propensity in the coding region are enriched for Regulation of Ras protein signal transduction and Src homology 3 (SH3) domain binding.
Temporal regulation of AgRP neurons mediates context-induced feeding
Authors: Reed Felicia , Dempsey Harry , Clarke Rachel E , Reichenbach Alex , Mequinion Mathieu , Stark Romana , Rawlinson Sasha , Foldi Claire J , Lockie Sarah H. , Andrews Zane B.
Published: February 2023

An environment can have a powerful influence over appetite and feeding behaviour. For example, an environmental context, which reliably predicts food, will increase the appetitive food drive to the same environment context. Interestingly, mice are required to be hungry to develop such a context-induced feeding (CIF) response, suggesting the neural circuits sensitive to hunger play an important role to associate an internal energy state with a particular environment context. Hunger-sensing Agouti related peptide (AgRP) neurons are activated by circulating signals of energy deficit and reset to a silenced state by gut feedback mechanisms following food consumption. We hypothesised that AgRP neurons are both necessary and sufficient to drive CIF in the absence of hunger. While fasting increased CIF, chemogenetic inhibition of AgRP neurons during context acquisition prevented this effect. Intriguingly, chemogenetic activation of AgRP neurons during context acquisition did not increase CIF, suggesting precise temporal firing properties may be required. Indeed, photostimulation of AgRP neurons, only during context exposure (ON-OFF in context), increased CIF. Moreover, AgRP photostimulation prior to context exposure, coupled with the termination of photostimulation in the context in the absence of food consumption, was sufficient to drive a subsequent CIF. Our results suggest that AgRP neurons regulate the acquisition of CIF when the temporal firing properties are matched to context exposure. These results further highlight that acute AgRP inhibition is a salient neural event underscoring the effect of hunger on associative learning.
The combined impact of killer whale consumptive and non-consumptive effects on northern sea otter population viability in the Western Aleutians Archipelago, Alaska
Authors: Tinker Tim , Schuette Paul , Weitzman Benjamin , Eisaguirre Joseph , Beatty William
Published: February 2023

Predators can alter the abundance, distribution, and behavior of prey populations through consumptive and non-consumptive effects. In the Aleutian Archipelago of Alaska, killer whales (Orcinus orca) are considered the most probable cause of northern sea otter (Enhydra lutris kenyoni) population declines in the southwestern Alaska stock, which led to their listing as a threatened distinct population segment under the Endangered Species Act. Much of the research attention in the Aleutian Archipelago region has focused on the consumptive effects of killer whales on sea otter population dynamics. Here, we explore non-consumptive effects by accounting for restricted sea otter habitat use within discrete predation refuges characterized by areas of shallow, complex reef habitats close to shore. We constructed Population Viability Analysis (PVA) models that incorporated sea otter count data collected by aerial and skiff-based methods over six decades (1959 – 2021) to inform uplisting (to endangered) and downlisting (delisting from ESA) criteria. Our models incorporated both density-dependent effects and density-independent effects on sex and age structure, which we termed predation hazards. Prior to 1990, predation hazards were negligible, fluctuated at high values between 1990 and the early 2000s, and then declined as sea otter populations reached low densities. We estimated a current regional abundance of 2,405 sea otters (95%CI = 1,734 – 3,238) in the Western Aleutians Management Unit. Our base PVA model that considered only inter-island fragmentation indicated the risk of the regional sea otter population becoming endangered was <5% when there were at least 1,500 otters (95%CI = 1,200 – 2,100), and provided a delisting threshold of 2,100 sea otters. A PVA model that accounted for restricted habitat use of sea otters within discrete predation refuges (i.e. inter-island and intra-island fragmentation) indicated a less encouraging potential for sea otter recovery. The probability of the population becoming endangered increased to >10% and the delisting threshold increased to >10,000 sea otters (nearly 5x higher). Our results indicate sea otters within fragmented predation refuges could be more susceptible to the effects of stochastic processes with potentially limited ability for rescue effects. Overall, our research reveals the importance of evaluating both consumptive and non-consumptive effects when considering conservation and management plans for at-risk populations thought to be limited by a predator.
Diversity of ribosomes at the level of rRNA variation associated with human health and disease
Authors: Rothschild Daphna , Susanto Teodorus Theo , Sui Xin , Spence Jeffrey P. , Rangan Ramya , Genuth Naomi R. , Sinnott-Armstrong Nasa , Wang Xiao , Pritchard Jonathan K. , Barna Maria
Published: February 2023

Ribosomal DNA and RNA (rDNA and rRNA) sequences are usually discarded from sequencing analyses. But with hundreds of copies of rDNA genes it is unknown whether they possess sequence variations that form different types of ribosomes that affect human physiology and disease. Here, we developed an algorithm for variant-calling between paralog genes (termed RGA) and compared rDNA variations found in short- and long-read sequencing data from the 1,000 Genomes Project (1KGP) and Genome In A Bottle (GIAB). We additionally developed a novel protocol for long-read sequencing full-length rRNA (RIBO-RT) from actively translating ribosomes. Our analyses identified hundreds of rDNA variants, most of which, surprisingly, are short insertion-deletions (indels) and dozens of highly abundant rRNA variants that are incorporated into translationally active ribosomes. To visualize variant ribosomes at the single cell level, we developed anin-siturRNA sequencing method (SWITCH-seq) which revealed that variants are co-expressed within individual cells. Strikingly, by analyzing rDNA, we found that variants assemble into distinct ribosome subtypes. We discovered that these subtypes acquire different rRNA structures by successfully employing dimethyl sulfate (DMS) probing of full length rRNA. With this atlas we investigated rRNA variation changes across human tissues and cancer types. This revealed tissue-specific rRNA subtype expression in endoderm/ectoderm-derived tissues. In cancer, low abundant rRNA variants can become highly expressed, which suggests the presence of cancer-specific ribosomes. Together, this study identifies and comprehensively characterizes the diversity of ribosomes at the level of rRNA variants which is dominated by indel variants, their chromosomal location and unique structure as well as the association of ribosome variation with tissue-specific biology and cancer.