bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of autophagy in prostate cancer cells stimulates Tau accumulation and aberrant mitotic spindle
Authors: Martellucci Stefano , Clementi Letizia , Colapietro Alessandro , Sabetta Samantha , Mattei Vincenzo , Angelucci Adriano
Published: April 2020

INTRODUCTION

Autophagy is a self‐degrading adaptation mechanism in which cytoplasmic macromolecules and organelles are delivered to the lysosome for degradation and recycling. In a physiologic context autophagy could be both a cytoprotective driver and a programmed death facilitator. Sustained autophagy has been frequently detected in neoplastic transformation, even though its promoting role in cancer growth is not fully elucidated. In recent years, a significant correlation has emerged between molecular dysfunction leading to Tau protein accumulation and interference with mitosis progression. In our study we investigated the potential association between autophagy and Tau accumulation and its role during cell mitosis.

METHODS

We evaluated in vitro the role of autophagy in modulating the expression of Tau protein in prostate cancer cell lines, analyzing morphological and molecular consequences during mitosis.

RESULTS

Tau protein was expressed at low level in prostate cancer lines and western blot analysis showed in these cells the presence of several phosphorylated and oligomeric forms. The pharmacologic inhibition of autophagy by treatment with chloroquine, induced in cancer cells a reduction in proliferation with an increment of number of cells in G2/M phase of cell cycle. In addition, in these cells we observed an accumulation of Tau protein, with an increased expression of both phosphorylated and oligomeric forms. Immunofluorescence analysis of untreated cells revealed that Tau was expressed mainly in dividing cells where it was localized on mitotic spindle. Inhibition of autophagy determined an evident upregulation of Tau signal in dividing cells with a diffuse protein localization in cytoplasm. The accumulation of Tau protein was associated with the presence of aberrant mitotic spindles, with monoastral spindle as major unusual figure.

CONCLUSIONS

Our data indicate that autophagy could exert a promoting role in cancer growth facilitating degradation of Tau protein and thus blocking the inhibitory effect of accumulated Tau forms on mitosis. Thus, the understanding of molecular mechanisms underlying the homeostatic control of Tau protein degradation during cell mitosis could represent an important goal also in oncology research.
PHARMACOLOGICAL EVALUATION OF BETA‐HYDROXIPHOSPHOCARNITINE IN NO‐ALCOHOLIC ESTEATOHEPATITIS INDUCED IN RATS
Authors: Sánchez-Quevedo Janet , Reyes-Esparza Jorge , Duarte-Vazquez Miguel Angel , Rodriguez-Fragoso Lourdes
Published: April 2020

Objective

To evaluate the pharmacological effect of Β‐Hydroxyphosphocarnitine (β‐HFC) on non‐alcoholic steatohepatitis induced in rats.

Introduction

Non‐alcoholic fatty liver disease (EHGNA) is a disease with high prevalence, usually accompanied by metabolic syndrome. Currently the treatment to EHGNA is pioglitazone. However, it is reported that pioglitazone causes several side effects reason to stop treatment. L‐carnitine has pharmacological effect in the metabolic syndrome, but it is poorly absorbed. The β‐HFC is an analogue of L‐carnitine that has shown effect on metabolic síndrome and maintaining its pharmacological properties.

Method

EHGNA was developed in male Wistar rats with a mixed diet based on coconut oil, carbon tetrachloride and high fructose drink. The treatments were as follows: Without treatment, pioglitazone and β‐HFC in healthy groups and with EHGNA. Histological sections of the liver were made and stained with Masson's Trichromic Stain to see collagen fibers and Hematoxylin & Eosin to see hepatocyte morphology. Quantities of liver enzymes Aspartate aminotransferase (AST), alanine aminotransferase (ALT), Gamma glutamyl transpeptidase (GGT) and alkaline phosphatase (ALP) and triglycerides were performed.

Results

In the histology of the healthy control rats, the hepatocyte architecture is well observed, no liver damage characteristics are observed, for example: fat drops, inflammatory cells, necrosis, apoptosis and hepatocytes in the balloon. Healthy groups administered with pioglitazone and β‐HFC as well as the control show no characteristics of liver damage. Histologies of rats with EHGNA showed the characteristics of liver damage and greater amount of collagen fibers. Rats with steatophepatitis treated with β‐HFC showed reduced hepatocyte damage and fibrosis compared to EHGNA control rats. Liver enzyme levels increased 2.5 times more in the steatohepatitis group compared to those in control rats. Liver enzyme levels showed considerable reduction in those rats with statohepatitis and were treated with β‐HFC, showing values for the enzymes ALT, AST, GGT and ALP 85.79, 101.3, 3.2 and 556.4 U/L, respectively for group with steatohepatitis and 74.34, 77.7, 1.0 and 327 U/L, respectively for the group with steatohepatitis treated with β‐HFC. Triglyceride levels increased in rats with steatohepatitis compared to those in control rats and reduction in levels was observed for those who were treated with β‐HFC showing values of 63.7 (control), 255.1 (steatohepatitis) and 48.4 mg/dL (steatohepatitis + β‐HFC). The same behavior showed liver levels 0.04, 0.14 and 0.04 mg/dL respectively.

Conclusion

B‐HFC reduced liver damage levels in rats with EHGNA.
Model‐Based Analysis of Lower Thoracic High‐Frequency Spinal Cord Stimulation (HF‐SCS) to Restore Effective Cough
Authors: Zander Hans J. , Kowalski Krzysztof E. , DiMarco Anthony F. , Lempka Scott F.
Published: April 2020

Background

Spinal cord stimulation (SCS, 50 Hz) is a useful method to restore an effective cough in persons with spinal cord injury (SCI). In animal models, dorsal HF‐SCS (~500 Hz) produces similar level of expiratory muscle activation with much lower stimulus amplitudes. Currently, the mechanism of this behavior is unknown.

Objective

The aim of the present study was to develop a computational model of lower thoracic SCS to investigate the mechanisms of the expiratory muscle activation via HF‐SCS.

Methods

We created a computer model of an anatomically‐accurate finite element model (FEM) of the lower thoracic (T9) canine spinal cord and surrounding anatomy with an implanted SCS electrode array. This model consisted of two main components: 1) multicompartment cable models of dorsal afferent fibers and 2) ventral motor axons. Model analysis allowed calculation of the extracellular voltages generated during SCS and the corresponding activation thresholds of each fiber type.

Results

In our simulations, we calculated the activation thresholds for several electrode combinations and stimulation frequencies (50 Hz, 500 Hz). As expected, the activation thresholds of the ventral motor axons were significantly higher than the activation thresholds of dorsal root axons. Therefore, low‐frequency SCS likely functions through the direct activation of ventral motor axons, while high‐frequency SCS operates via recruitment of myelinated afferent fibers at significantly lower stimulation amplitudes. In our simulations, the stimulation frequency did not significantly alter activation thresholds in dorsal or ventral axons.

Conclusions

Our results suggest that both high‐ and low‐frequency SCS directly activate dorsal rootlets at low amplitudes (~1 mA); however, the trans‐synaptic spinal circuits responsible for eliciting cough are only effective at higher stimulation frequencies. Consequently, low‐frequency stimulation results in effective cough by bypassing this circuitry entirely to directly activate the ventral motor axons at the cost of requiring higher stimulation amplitudes. In future work, we will perform simulations directly assessing the ability of SCS to engage these spinal circuits. We will compare our model results to experimental measurements in an animal model of SCS to produce cough.

Disclosure

Dr. DiMarco holds two U.S. patents for technology related to the content of this abstract: Method and Apparatus for Electrical Activation of the Expiratory Muscles to Restore Cough (5,999,855) and Bipolar Spinal Cord Stimulation to Activate the Expiratory Muscles to Restore Cough (8,751,004).

Support or Funding Information

Merit Review Award I01‐RX‐001488 from the U.S. Department of Veterans Affairs, Rehabilitation Research and Development Service.
Methylphenidate Does Not Hypersensitize the Rat Heart to Ischemic Injury
Authors: Rorabaugh Boyd R. , Seeley Sarah L. , Stoop Thorne S. , D'Souza Manoranjan S.
Published: April 2020

Background

Methylphenidate is commonly used for the treatment of attention deficit hyperactivity disorder. The cardiovascular safety of methylphenidate has been a subject of debate with some studies indicating that methylphenidate increases the likelihood of experiencing a myocardial infarction. However, it is unknown whether methylphenidate alters the extent of myocardial injury during an ischemic insult. Previous work demonstrated that a history of methamphetamine exposure increases infarct size and postischemic recovery of contractile function in the ischemic heart in a sex‐dependent manner. Methylphenidate and methamphetamine both increase synaptic concentrations of dopamine and norepinephrine, resulting in enhanced adrenergic and dopaminergic signaling. However, it is unknown whether methylphenidate also hypersensitizes the heart to ischemia. The purpose of this study was to determine whether methylphenidate increases the extent of myocardial injury during an ischemic insult.

Methods

Male and female rats were treated with methylphenidate (5 mg/kg/day) or saline for 10 days. Hearts were subjected to a 20 min ischemic insult and 2 hours of reperfusion on a Langendorff isolated heart apparatus on day 11. Cardiac contractile function was monitored via an intraventricular balloon, and myocardial injury was assessed by triphenyltetrazolium chloride staining.

Results

Methylphenidate significantly increased locomotor activity in male and female rats. Male hearts had significantly larger infarcts than female hearts, but methylphenidate had no impact on infarct size or postischemic recovery of contractile function in hearts of either sex.

Conclusion

These data indicate that methylphenidate does not increase myocardial sensitivity to ischemic injury.

Support or Funding Information

This work was supported by 1R15HL145546 to BRR.
Scarring and Arrhythmia in the Aged Infarcted Heart: The Role of Senescent Fibroblasts
Authors: Baggett Brett , Murphy Kevin , Cao Yueming , Turan Nilufer N. , Lu Yichun , Schofield Lorraine , Kim Tae Yun , Terentyev Dmitry , Sedivy John , Choi Bum-Rak , Koren Gideon
Published: April 2020

Arrhythmogenic tissue remodeling after myocardial infarction (MI) has been well described in young animals, but how this wound healing process changes with age and leads to increased risk of sudden cardiac death (SCD) post‐MI is not well understood. Cellular senescence is an irreversible cell cycle arrest process which regulates tumor suppression and wound healing, however accumulation of senescent cells in various tissues is thought to drive aging pathology. We hypothesize that with age, persistent cardiac myofibroblast (CMF) senescence promotes persistent inflammation in the scar and thereby facilitating arrhythmogenesis. Using the New Zealand White Rabbit as a model of the aging heart, we show that aged infarcted rabbits display increased peri‐procedural arrhythmic death. Optical mapping studies at 3‐ and 12‐weeks post MI revealed prolonged action potential duration (APD) in the infarct border zone (IBZ) as compared to the remote zone (RZ) in aged rabbits while in young rabbits IBZ’s APD was shorter than RZ’s APD. We also observed slowed conduction and APD alternans at the IBZ which correlated with anchoring of higher frequency ventricular fibrillation in the IBZ of aged rabbits. We show no change in scar size or geometry between young and aged rabbits over the first 12 weeks post‐MI. However, we observed delayed and persistent CMF senescence in the aged scar, even until 12 weeks post‐MI. Using immunohistochemistry and immunofluorescence, we show most senescent cells in the scar are myofibroblasts. We also demonstrate increased expression of senescence‐associated markers in the aged IBZ and scar compared to young, and colocalization of senescent CMFs and inflammatory macrophages in the aged scar and IBZ. Finally, we observe an increase in circulating inflammatory factors from aged rabbit blood samples compared to young, some of which further increased post‐MI. In conclusion, we demonstrate the infarcted aged heart heals differently than young, including delayed and persistent scar CMF senescence, increased VT/VF, and electrophysiological remodeling. Our data support our hypothesis that altered CMF senescence with age induces pathological tissue remodeling post‐MI, underlying the increased risk of SCD.

Support or Funding Information

The authors would like to acknowledge our supporting NIH funding sources, R21 grant 1R21AG049608‐01 and R01 grant R01HL139467
Consumption Pattern of Fruits and Vegetables among solar market garden women’s groups of Kalalé district of Northern Benin
Authors: Valenti Michelle , Alaofè Halimatou
Published: April 2020

BACKGROUND

Anemia, iron deficiency (ID) and vitamin A deficiency (VAD) continue to be significant public health issues in northern Benin despite the installation of solar market gardens (SMGs), suggesting an urgent need of implementing nutrition programs to benefit the full impact of SMGs. One way to enhance the benefit is to know which fruits and vegetables (F&V) are currently consumed to determine their suitability to promote as a crop.

OBJECTIVE

To assess the consumption pattern of F&V in the Kalalé district of Northern Benin in order to identify which F&V could be promoted for improving anemia and micronutrients status.

METHODS

A cross‐sectional survey was conducted among 250 mothers/caregivers aged 19 years and above. Questionnaires were used to assess socio‐demographic characteristics and types of staples foods consumed including the types of F&V consumed and their frequency of consumption. Descriptive analyses were tabulated using STATA version 14.

RESULTS

Several F&V are eaten in the district, but their frequency of daily consumption is low. However, mangoes are abundant during the two fruiting seasons and could be easily accessed by the participating women. Likewise, Moringa oleifera leaves are available all year round and could be incorporated into selected recipes.

CONCLUSIONS & IMPLICATIONS

Mangoes and Moringa oleifera leaves are potential sources of β‐carotene to be promoted on a sustainable basis among SMG women’s groups of Kalalé district of Northern Benin. Additionally, nutrition educational messages should include behavior change campaigns that will make the women see the need for consuming F&V on a daily basis.

Support or Funding Information

University of Arizona Mel and Enid Zuckerman College of Public Health Dean’s Annual Fund University of Arizona Graduate & Professional Student Council Travel Grant
Acute blood glucose and vascular endothelial responses to passive heating in individuals with type 2 diabetes
Authors: Behzadi Parya , Gravel Hugo , Ravanelli Nicholas , Gagnon Daniel
Published: April 2020

Introduction

Insulin resistance and hyperglycemia predispose individuals with type 2 diabetes mellitus (T2DM) to endothelial dysfunction and a greater risk of cardiovascular diseases. Numerous animal studies suggest that heat exposure improves glucose control and human studies suggest that it improves markers of vascular health. However, the physiological responses to heat exposure remain understudied in individuals with T2DM, particularly within a therapeutic context.

Objective

To characterize blood glucose and vascular endothelial responses to heat exposure in T2DM and determine if heating acutely improves glucose control and endothelial function in this population.

Methods

On separate randomized days, six adults (2 males/4 females, 60 ± 12 years) with T2DM (≥1 year, HbA1c: 7.0 ± 0.5%, fasting glucose: 7.79 ± 1.13 mmol/L) were immersed in control (34°C, 90 minutes) or hot (40–41°C, core temperature ≥38.5°C for 60 minutes) water. Participants were fasted (12 hours) and withheld anti‐diabetic medication (≥24 hours). Blood glucose and insulin as well as brachial artery shear rates were measured at baseline, during immersion and 45–60 minutes after immersion. Brachial artery flow‐mediated dilation (FMD) was measured at baseline and 45 minutes after immersion. Blood glucose control was evaluated during an oral glucose tolerance test (OGTT, 75 g) 60 minutes after immersion. Core (Tcore, rectal) and mean skin (Tskin) temperatures, heart rate (HR), systolic (SBP) and diastolic (DBP) blood pressures were measured throughout the protocol.

Results

Compared to control, hot water immersion increased Tcore (+1.44°C [1.05, 1.83], P<0.01), Tskin (+2.36°C [1.21, 3.51], P<0.01), HR (+35 bpm [26, 43], P<0.01), and decreased DBP (−7 mmHg [−2, −13], P<0.01) and SBP (−9 mmHg [−1, −16], P=0.03). Hot water immersion increased antegrade (+71 s−1 [35, 107], P<0.01) and decreased retrograde (−15 s−1 [0.9, 3], P=0.06) shear rates. Nonetheless, FMD was unchanged during both control (−0.74% [−4.33, 2.86], P=0.81) and hot (−1.93% [−5.53, 1.66], P=0.30) water immersion. Blood glucose decreased progressively throughout the control protocol (−1.3 mmol/L [−0.8, −1.9], P<0.01), whereas hot water immersion attenuated this decrease (+0.1 mmol/L [−0.5, +0.6], P=0.93). Hot water immersion did not alter glucose (P=0.17) and insulin (P=0.99) area under the curve during the OGTT.

Conclusion

These preliminary results suggest that the physiological responses associated with heat exposure do not translate into acute improvements in blood glucose control or brachial artery endothelial function in individuals with T2DM.

Support or Funding Information

Cardiometabolic Health, Diabetes and Obesity Research Network and Diabetes Quebec.
Glutamine Synthetase and Cytochrome P‐4502E1 Characterize Metabolic Zonation in Human Liver: Immunohistochemical Study
Authors: Mak Ki Mark
Published: April 2020

Hepatic metabolic zonation endows metabolic homeostasis and proper functioning of the liver under various physiological conditions. The zonation reflects the heterogeneity of gene expression and functions of hepatocytes along the centro‐portal axis of the liver lobule. The Wnt/ß‐catenin signaling pathway is the master regulator of hepatic zonation. The pathway maintains the metabolic zonation of glutamine synthetase (GS) and cytochrome P‐4502E1 (2E1) —among others—in the centrilobular parenchyma. GS detoxifies ammonia entering the liver as well as serves as an immunohistological marker of perivenous hepatocytes that make up the first three cell layers around the central vein. 2E1 mediates metabolism of xenobiotics and has a broader centrilobular expression than that of GS. While the zonation of GS and 2E1 has been documented in animal models, the phenomenon remains to be evaluated in human liver. This study assessed expression of GS and 2E1 in human cadaveric livers by immunohistochemistry. Ten livers of cadavers (mean age, 79.9 years) with minimal fibrotic changes were studied. Consecutive paraffin liver sections were stained with a polyclonal GS antibody or 2E1 antibody using an immunoperoxidase method. The central vein was defined as a hepatic vein located at the center of the classic liver lobule and showed a cross‐sectional diameter not exceeding 150 μm. In each liver, five to six veins were examined for the distribution of GS+ hepatocytes in the centrilobular area. In most cases, GS+ hepatocytes were arranged in a band of 1 to 3 cells wide in the perivenous area around the central veins (perivenous cells). In some instances, GS staining was observed in hepatocytes beyond the perivenous area (recorded as non‐perivenous cells). Midlobular and periportal hepatocytes lacked GS staining. Expressed as per central vein, counts of perivenous GS+ cells were 14.9 ± 2.2 and non‐perivenous cells were 5.6 ± 1.8. The mean 2E1 zonal score was 1.35 ± 0.35. Total GS+ cell numbers (perivenous plus non‐perivenous) were found to correlate with 2E1 zonal scores (P > 0.05, N = 10).

Conclusions

Expression of GS and 2E1 characterizes the metabolic zonation in the liver of aged cadavers, in accordance with the findings of animal studies. The correlation of GS with 2E1 expression suggests a link between the hepatic zonation of GS and 2E1, which is consistent with the notion that the zonation is mediated by the Wnt/ß‐catenin signaling pathway.
Integrating Clinical Imaging into Anatomy Curriculum to Increase Clinical Preparedness
Authors: Altshuler Paulina , Tchen-Tomasino Emily , Jorgensen Mike
Published: April 2020

The anatomy curriculum at Rocky Vista University College of Osteopathic Medicine (RVUCOM) has historically placed emphasis on cadaver‐based lectures and laboratory sessions with little lecture time devoted to radiology. Beginning with the Class of 2020, we implemented an additional “Integrated Anatomy” lecture series with eleven lectures detailing normal anatomy on radiographs, CTs, and MRIs during the first year and six review lectures with board‐style practice questions during the second year. The goal of this lecture series was to increase exposure to radiology while also increasing students’ confidence interpreting imaging studies. Students’ perceived confidence was evaluated with anonymous surveys using the Likert scale, and were analyzed with the Kruskal‐Wallis test to assess our hypothesis that increasing pre‐clinical radiology exposure will increase confidence interpreting images and overall preparedness for second‐year courses and third‐year clerkships. We found statistical significance to support our hypothesis that students who received the “Integrated Anatomy” lecture series felt more prepared for second‐year courses, third‐year clerkships, identifying anatomical structures on radiographs, and identifying abnormalities on MRIs.
Cadherin‐3 and Catenin‐α2 are novel MMP‐12 substrates
Authors: Chalise Upendra , Lindsey Merry L.
Published: April 2020

Adverse cardiac remodeling after myocardial infarction (MI) is a major cause of heart failure. Degradation of the existing extracellular matrix (ECM) and subsequent formation of a new ECM scar are prominent components of cardiac remodeling. Matrix metalloproteinases (MMPs) increase after MI to dictate the degradation component. MMP‐12 inhibition after MI worsens dilation of the left ventricle by increasing its wall thinning, suggesting a possible protective role for MMP‐12 in MI remodeling. We hypothesized that identifying MMP‐12 substrates could provide mechanistic insight into MI roles. We used the Mouse Heart Attack Research Tool 1.0 (mHART 1.0) database and performed regression analysis to identify ECM genes that correlated with MMP‐12 expression. Of the 83 ECM genes compared to MMP‐12, 41 showed significant regressions (all p<0.05). This indicates a strong relationship between ECM breakdown after MI and MMP‐12. Synaptotagmin‐1, Contactin‐1, MMP‐10, MMP‐13, and Catenin‐α2 were the top 5 genes with significant positive correlation (Pearson coefficient r>0.99 and p<0.0001 for all). By Enrichr analysis for transcription factor pathway exploration, Yap1 signaling had a top combined score of 9.37 and p‐value 0.0033, indicating the upregulation of HIPPO signaling as the most enriched pathway. Catenin‐α2(Ctnna2), Cadherin‐1 (Cdh1), and Cadherin‐3 (Cdh3) were the three genes linked to Yap1‐ Hippo signaling. By substrate cleavage assay using 0.1 μg MMP‐12 enzyme to 0.5 μg substrate, Cdh3 and Ctnna2 but not Cdh1 were identified as novel MMP‐12 substrates. Cdh3 and Ctnna2 showed robust cleavage at 15 min, and Ctnna2 was completely processed by 3h. Our combined results reveal that MMP‐12 actions in MI remodeling could be attributed in part to Cdh3 and Ctnna2 proteolytic processing.

Support or Funding Information

We acknowledge funding from the National Institutes of Health under Award Numbers HL075360, HL129823, and HL137319, and the Biomedical Laboratory Research and Development Service of the Veterans Affairs Office of Research and Development under Award Numbers 5I01BX000505.
a‐arrestin ARRDC3 is a Multifunctional Adaptor That Regulates G Protein‐Coupled Receptor Signaling and Breast Cancer Invasion
Authors: Pan Wen-An , Arakaki Aleena , Wedegaertner Helen , Kufareva Irina , Trejo JoAnn
Published: April 2020

Aberrant G protein‐coupled receptor (GPCR) signaling drives cancer initiation and progression, but the mechanism underlying dysregulated GPCR signaling is unknown and represents a major gap in knowledge. Protease‐activated receptor‐1 (PAR1) is proteolytically activated and requires lysosomal degradation for signal termination. PAR1 overexpression correlates with high tumor grade and poor prognosis in breast cancer patients. Increased PAR1 expression results from altered receptor trafficking, which causes persistent signaling and increased invasiveness. We recently discovered that loss of the tumor suppressor α‐arrestin domain‐containing protein‐3 (ARRDC3) is responsible for dysregulated PAR1 trafficking, persistent signaling and breast cancer invasion. This work establishes for the first time a link between ARRDC3 and GPCR dysfunction in cancer. However, it is not known how ARRDC3 alters the dynamics of PAR1 signaling via receptor trafficking or if other determinants are important. We found that ARRDC3 regulates PAR1‐induced non‐canonical endosomal p38α MAPK signaling, but not PAR1‐rapid initiation of Gα12/13‐RhoA signaling from the plasma membrane. Interestingly, ARRDC3 interacts with NEDD4 ubiquitin E3 ligases via C‐terminal PPXY motifs, which is required for agonist‐induced PAR1 degradation, but not essential for regulation of p38α MAPK signaling. However, deletion of the C‐terminus tail abrogated both processes, suggesting that an additional determinant within the C‐terminustail distinct from the PPxY motifs is also important for mediating ARRDC3 function. In addition, PAR1‐stimulated invasion is dependent on p38a MAPK and regulated by ARRDC3, which requires the C‐terminal domain but is only partially suppressed by ARRDC3 PPXY mutant. We further discovered a novel phosphorylated Ser/Thr cluster in the C‐terminus that is critical for ARRDC3 function including modulation of ARRDC3 homo‐oligomerization and basal conformation. Meta‐analysis of breast cancer patients indicates that ARRDC3 and genes involved in PAR1‐activated p38α MAPK are associated with poor prognosis in the aggressive subtype of breast cancer patients. Currently, we are determining the molecular mechanisms by which phospho‐Ser/Thr cluster effects ARRDC3 conformation and function. In conclusion, this work has provided critical insight that improves our understanding of ARRDC3 function in spatiotemporal regulation of GPCRs signaling in invasive breast carcinoma, which may provide new therapeutic options for treating metastatic breast cancer.

Support or Funding Information

NIH/NIGMS R35 GM127121
Myeloid mineralocorticoid receptor activation alters metabolism in HFD‐salt fed mice
Authors: Lefranc Clara , Gabarre Paul , Genty Marie , Ramirez Roberto Palacios , Feraco Alessandra , Caprio Massimiliano , Jaisser Frederic
Published: April 2020

Context and Objectives

In obese mice and patients, mineralocorticoid receptor (MR) activation has deleterious consequences on metabolic and cardiovascular parameters. In mice, adipose tissue MR triggers mitochondrial and vascular dysfunction. Adipocyte MR activation induces metabolic and vascular dysfunction in a context of obesity, however, obese adipose tissue is also infiltrated with macrophages, which are capable of expressing MR. Thus, we hypothesised that macrophage MR could also be responsible for metabolic and cardiovascular complications in high‐fat high‐salt diet‐fed mice.

Methods

We fed myeloid cell‐specific MR knock‐out male and female mice (My‐MRKO) with a 6‐week high fat – high salt diet (60% kcal from fat, 1.2% salt in drinking water) and studied weight gain, whole body and specific organs composition by nuclear magnetic resonance and insulin sensitivity to explore the metabolic phenotype. Blood pressure measurements by plethysmography and vascular reactivity studies by wire myography were also performed to characterise the cardiovascular phenotype.

Results

My‐MRKO male mice under high fat – high salt diet displayed reduced weight (37.8 g [interquartile range 34.1–40.8] vs 42.5 g [39.5–45.2]; P = 0.0128), preserved muscle mass (61.5 % [55.8–63.3] vs 56.8 % [54.1–58.5]; P = 0.0337) and a strong tendency to decreased fat mass (26.1 % [23.7–33.9] vs 31.7 % [27.1–35.4]; P = 0.0728). Moreover, liver fat content at sacrifice was decreased (5.8 % [2.1–9.1] vs 9.3 % [5.9–13.4]; P = 0.0491). Those effects were not observed in female mice. Fasting glycaemia was decreased in male My‐MRKO mice (9.2 mmol/L [6.4–12.8] vs 13.2 mmol/L [10.4–16.1]; P = 0.0005) and insulin resistance was decreased in both gender (area under curve 9915 [9398–10043] vs 11550 [9795–12848]; P = 0.0458 in males, 8291 [7812–8884] vs 9544 [8918–9735]; P = 0.0177 in females), as assessed by an insulin tolerance test. A clear effect was observed on blood pressure only in female mice (blood pressure 96 mmHg [94–97] vs 106 mmHg [104–109]; P = 0.0004), whereas there was no modification in mesenteric arteries vascular reactivity in both gender.

Conclusions

Thus, our findings show that myeloid cell‐MR regulates systemic metabolic parameters, highlighting a new role in obesity and its associated complications for MR expressed in myeloid cells.

Support or Funding Information

Sorbonne Université, Inserm, CARMMA‐RHU
Design of first in class bitopic ligands targeting the sodium binding pocket in opioid receptors
Authors: Faouzi Abdelfattah , Zaidi Saheem , Che Tao , Kormos Chad , Zhang Tiffany , Madasu Manish , Eans Shainnel , Simons Chloe , Pan Ying-Xian , McLaughlin Jay , Carroll Ivy , Hassani Ream Al , Roth Bryan , Katritch Vsevolod , Majumdar Susruta
Published: April 2020

Opioid agonists have been used in the field of pain management for centuries, but are considered a major public health concern, especially in the US. One of the biggest challenges consists in understanding the ability to target specifically the different subtypes of opioid receptors and to trigger a specific functional response correlating with an in vivo analgesic response devoid of side‐effects.

These opioid receptors are part of the GPCRs family, which is known to have common features across different subfamilies. One important feature of these receptors is the presence of allosteric binding pockets, which is progressively gaining momentum as a novel target for current therapeutic strategies. We hypothesized that targeting an allosteric site could potentially allow for receptor functional selectivity, helping to reduce the overall side effects profile. One site, in particular, known as the Na+ binding pocket, is known to play a key role in GPCR function and is thought to be the center of the functional mechanism, with several bias switches characterized in the pocket residues (Katritch et al., Trends Biochem Sci. 2014).

Here we report structure‐based rational drug design of bitopic ligands targeting the Na+ pocket of mu (MOR) and kappa opioid receptors (KOR). Our designed bitopics show high affinity, subtype selectivity, G‐protein bias and analgesic actions in mice. Mutations in Na+ binding pocket diminished functional potency validating allosteric site targeting. Together, results suggest the possibility of achieving subtype selectivity and identification of an additional subpocket in both MOR and KOR to reduce βarrestin‐2 recruitment by targeting an allosteric pocket in the opioid receptors. The highly conserved constitution of the Na+ binding pocket opens the opportunity for rational discovery of new GPCR modulators with desired functional, and potentially therapeutic profile for opioids as well as other class A GPCRs.

Support or Funding Information

We gratefully acknowledge the financial support from Faculty Research Incentive Fund, NIDA, NIAAA and US department of defense. We also would like to thank the Philippe Foundation for their financial support.
Use of visual and performing arts to raise cultural awareness through obesity course within pharmacy education
Authors: Jungsuwadee Paiboon , Iwuchukwu Otito , Patel Shreya
Published: April 2020

Introduction

The 2016 Accreditation Council for Pharmacy Education Standards call for exposure to cultural factors within pharmacy curriculum. Current efforts to meet these standards using common pedagogical techniques have had some success; however, there is still a need to identify different ways of teaching and learning cultural competency. An obesity elective course offered in the third professional year used visual and performing arts as a student‐centered learning tool to raise cultural awareness and advocate for culturally inclusive treatment.

Research question

Can visual and performing arts be used as a student‐centered learning tool to raise cultural awareness and advocate for culturally inclusive treatment approaches?

Study design

Students were randomized into groups and charged with selecting one patient group at risk for weight based biases from a pre‐specified list. The groups were then required to use one form of visual and performing arts (e.g., song, print media, etc.) to raise cultural awareness and present culturally inclusive treatment options for their chosen at‐risk patient group.

Methods

The use of visual and performing arts was assessed on creativity, attractiveness, and content accuracy in the context of obesity. An online audience response system and a multiple choice quiz was utilized to assess student learning. Students’ perception of how well the activity met its intended aims was assessed through a survey questionnaire.

Results

Six groups consisting of 4 to 5 students completed the activity. Student scores on creativity, attractiveness and content accuracy ranged from 97.3% to 77.8%, with an average of 83.8%. On the quiz, 90% of students achieved a score of 85% or higher. For students’ perception, 86% of students agreed that the activity improved their understanding and 91% agreed the activity improved their awareness of cultural differences associated with weight biases.

Conclusion

Visual and performing arts can be used as a student‐centered learning tool to expose students to cultural disparities within pharmacy curriculum.
Activation of Autophagy Restores Nitric Oxide as the Primary Mechanism of Flow‐Mediated Dilation in the Absence of Telomerase Reverse Transcriptase Activity
Authors: Hughes William Edward , Gutterman David D. , Beyer Andreas M.
Published: April 2020

Arteriolar vasodilation to shear stress, termed flow‐mediated dilation, is endothelium‐dependent, primarily mediated by nitric oxide (NO) in health, and hydrogen peroxide (H2O2) in subjects with coronary artery disease (CAD). Previous evidence has demonstrated that autophagy and the catalytic subunit of telomerase, telomerase reverse transcriptase (TERT), govern the mediator of arteriolar FMD such that inhibition of telomerase activity or autophagy initiate a switch in mechanism of dilation from NO to H2O2 in non‐CAD, while activation of either telomerase or autophagy switch the mechanism of dilation from H2O2 to NO in CAD arterioles. We examined whether autophagy is necessary for the vascular effects of TERT utilizing a rat loss of function model that phenocopies the vascular changes observed in subjects with CAD. We hypothesized that activation of autophagy will switch the mechanism of dilation from H2O2 to NO, despite absence of TERT. Trehalose, an autophagy activator was administered in drinking water (2%) for 4 weeks. Mesenteric arterioles (3rd or 4th order) were isolated from TERT KO and wild‐type (WT) littermates and prepared for video and fluorescent microscopy. Control KO and WT animals received normal drinking water. Internal diameters were measured following increases in intraluminal flow (pressure gradient) in the presence/absence of a NOS inhibitor (L‐NAME; 100μM) or H2O2 scavenger (Peg‐Catalase, Peg‐Cat; 500 units/mL). Shear‐induced autophagy was examined using Lysotracker Red DND‐99 in response to sustained shear (100 cmH2O) for 1 hour. FMD is expressed as % maximal dilation, while Lysotracker data is expressed as relative fold change in fluorescent signal above static. In arterioles from untreated KO rats H2O2 is the primary mechanism of FMD (Vehicle: 64±9% vs. L‐NAME: 70±8% vs. Peg‐Cat: 16±4%; n=9; P<0.05), while NO predominates in WT arterioles (P<0.05). 4 weeks of 2% trehalose supplementation initiated a switch to NO as the primary mechanism of dilation in KO arterioles (Vehicle: 55±9% vs. L‐NAME: 19±6% vs. Peg‐Cat: 58±13%; n= 6–8; P<0.05). Similarly, in WT arterioles, NO is maintained as the primary mechanism of dilation (P<0.05). In KO rats shearinduced autophagy was reduced, while trehalose augmented the shear‐induced activation of autophagy in both KO and WT relative to untreated KO and WT animals (P<0.05). Our data demonstrate that trehalose initiates a switch from H2O2 to NO in KO rats, and augments shearinduced autophagy in both KO and WT. Taken together, autophagy is necessary for maintenance of NO as the primary mechanism of FMD in the absence of TERT activity.

Support or Funding Information

Funded by T32GM089586‐09 (WEH), R01‐HL135901‐02 (DDG), and R01‐HL133029‐02 (AMB)
Neuroanesthesia Practice During the COVID-19 Pandemic: Recommendations From Society for Neuroscience in Anesthesiology and Critical Care (SNACC)
Authors: Flexman Alana M. , Abcejo Arnoley S. , Avitsian Rafi , De Sloovere Veerle , Highton David , Juul Niels , Li Shu , Meng Lingzhong , Paisansathan Chanannait , Rath Girija P. , Rozet Irene
Published: April 2020

The pandemic of coronavirus disease 2019 (COVID-19) has several implications relevant to neuroanesthesiologists, including neurological manifestations of the disease, impact of anesthesia provision for specific neurosurgical procedures and electroconvulsive therapy, and health care provider wellness. The Society for Neuroscience in Anesthesiology and Critical Care appointed a task force to provide timely, consensus-based expert guidance for neuroanesthesiologists during the COVID-19 pandemic. The aim of this document is to provide a focused overview of COVID-19 disease relevant to neuroanesthesia practice. This consensus statement provides information on the neurological manifestations of COVID-19, advice for neuroanesthesia clinical practice during emergent neurosurgery, interventional radiology (excluding endovascular treatment of acute ischemic stroke), transnasal neurosurgery, awake craniotomy and electroconvulsive therapy, as well as information about health care provider wellness. Institutions and health care providers are encouraged to adapt these recommendations to best suit local needs, considering existing practice standards and resource availability to ensure safety of patients and providers.
Treatment of Tibial and Femoral Bone Defects With Bone Transport Over an Intramedullary Nail
Authors: Bas Ali , Daldal Fehmi , Eralp Levent , Kocaoglu Mehmet , Uludag Serkan , Sari Seckin
Published: April 2020

Objectives:

To evaluate the results of the bone transport over an intramedullary nail (BTON) technique for the treatment of segmental bone defects.

Design:

Retrospective review of case series.

Setting:

Level 1 trauma center.

Patients/Participants:

We included 40 patients who underwent reconstruction of the lower limb with BTON technique between 2000 and 2018. The technique was performed in the tibial segments in 21 patients and in the femoral segments in 19 patients.

Intervention:

The surgical technique was performed in 2 stages for patients with infected nonunion. Infection was eradicated in all patients at the first stage. For the BTON at the second stage, monolateral external fixators and circular external fixators were used for femoral and tibial defects, respectively. In cases of defects without any infection, debridement with a single-stage BTON was performed.

Main Outcome Measurements:

Complications as well as radiological and clinical results were evaluated according to the criteria of Paley–Maar.

Results:

Minor complications occurred in 11 patients: pin site problems (9), cellulitis (1), and skin detachment due to Schanz screw (1). Major complications occurred in 8 patients: docking site nonunion (4), early consolidation and Schanz screw failure (1), knee flexion contracture (1), and ankle equinus contracture (2). Four patients had osteomyelitis as residual sequelae. Bone score was excellent in 27 patients. Excellent functional results were obtained in 31 patients.

Conclusions:

The BTON technique is associated with low cost because of the short treatment period, low complication risk, and rapid rehabilitation and is not limited by the amount of bone transport.

Level of Evidence:

Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.
Clinical assessment of generalized ligamentous laxity using a single test: is thumb-to-forearm apposition enough?
Authors: Cahill Sean V. , Sharkey Melinda S. , Carter Cordelia W.
Published: April 2020

The complete Beighton criteria, commonly used to establish the diagnosis of generalized ligamentous laxity (GLL), include nine discrete examination maneuvers. However, busy examiners may perform only a single maneuver (e.g. passive apposition of the thumb to the forearm) as a rapid method of assessment. We hypothesize that the use of a single-joint hypermobility test does not reliably identify the presence of GLL. Healthy patients 2–18 years old presenting to a general pediatric orthopaedic clinic were screened for participation. Exclusion criteria included the presence of a systemic illness, neuromuscular disease, and inability to complete the examination. Subjects were assessed for GLL according to the Beighton criteria, using a cutoff score of 5. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and likelihood ratio were calculated for the thumb-to-forearm apposition test with the composite Beighton score used as the gold standard. Two hundred and four patients were included in the study, 111 females and 93 males, with an average age of 10.7 years. The prevalence of GLL was 13.3%. When thumb-to-forearm apposition was performed unilaterally, the PPV was poor (34%). Conversely, the NPV was excellent (99%). Sensitivity of thumb-to-forearm motion was extremely high (99%), although the specificity of this test was modest (67%). The likelihood ratio was fair (+3.3). Performing the test bilaterally did not significantly change its utility. Thumb-to-forearm apposition testing was equally effective in identifying the presence of GLL in males and females. When performed in isolation, assessment of thumb-to-forearm apposition has a poor PPV, excellent NPV, and modest likelihood ratio. It is an extremely sensitive test, with only fair specificity. Other upper extremity tests of GLL perform similarly. Therefore, while single tests like thumb-to-forearm apposition may be helpful for ‘ruling out’ GLL, they are less reliable at identifying it correctly. When single tests are used to identify GLL in either clinical or research settings, the results should be interpreted with caution. Level of evidence Level I (diagnostic).
Serial Casting in Neuromuscular and Syndromic Early-onset Scoliosis (EOS) Can Delay Surgery Over 2 Years
Authors: LaValva Scott , Adams Alexander , MacAlpine Elle , Gupta Purnendu , Hammerberg Kim , Thompson George H. , Sturm Peter , Garg Sumeet , Anari Jason , Sponseller Paul , Flynn John , Cahill Patrick J.
Published: April 2020

Background:

The primary goal in managing early-onset scoliosis (EOS) is delaying/preventing surgical intervention while allowing improved spinal growth and chest wall and lung development to improve life expectancy. The effectiveness of serial casting for patients with neuromuscular and syndromic EOS is unclear.

Methods:

Patients from 2 multicenter registries who underwent serial casting for nonidiopathic scoliosis (NIS) were reviewed retrospectively. Comparisons were made between precasting and postcasting major and compensatory curves and spine height. The need for surgical intervention and any treatment complications were documented. Risk factors for major curve progression from baseline to casting cessation were evaluated via univariate analysis.

Results:

Forty-four patients (23 females; 21 males) with NIS (26 syndromic, 18 neuromuscular) and a mean age of 3.2 years at baseline were included. Mean follow-up and casting duration was 3.9 and 2.0 years, respectively. There were no statistically significant differences between mean precasting and postcasting major curve (55 vs. 60 degrees; P=0.348), minor curve (31 vs. 33 degrees; P=0.510), or rib-vertebra angle difference (18 vs. 29 degrees; P=0.840). However, thoracic height (15.5 vs. 16.8 cm; P=0.031) and lumbar height (8.9 vs. 9.8 cm; P=0.013) were significantly greater upon casting cessation. Currently, 13 patients (30%) have had successful casting (improvement of major curve ≥10 degrees) while 24 patients (55%) experienced major curve progression (worsening), and 19 patients (43%) required surgical intervention. Mean time from first casting to surgery was 34.5±15.1 months. There were no statistically significant predictors for major curve progression on univariate analysis.

Conclusions:

Spinal deformity progression despite casting and the subsequent need for surgical intervention for NIS were significantly higher compared with those reported for idiopathic EOS. However, serial casting did afford a substantial delay in surgical intervention. Ultimately, serial casting for neuromuscular or syndromic EOS is an effective strategy for delaying surgical intervention, despite suboptimal radiographic outcomes.

Level of Evidence:

Level III.
Alterations of B Cells in Immunosuppressive Phase of Septic Shock Patients*
Authors: Dong Xijie , Liu Qinxin , Zheng Qiang , Liu Xinghua , Wang Yuchang , Xie Zhenxing , Liu Tao , Yang Fan , Gao Wei , Bai Xiangjun , Li Zhanfei
Published: April 2020

Objectives:

Septic shock is a subset of sepsis related to acute circulatory failure characterized by severe immunosuppression and high mortality. Current knowledge about B-cell status in the immunosuppressive phase of septic shock is sparse. The aim of this study was to investigate the alterations of B Cells in the immunosuppressive phase of septic shock.

Design:

Prospective cohort study.

Setting:

Adult ICUs at a university hospital.

Patients:

Adult septic shock patients without any documented immune comorbidity.

Interventions:

None.

Measurements and Main Results:

The absolute counts of lymphocytes and B cells of 81 patients and 13 healthy controls, and serum immunoglobulin levels of 64 patients and 10 healthy controls were determined by clinical laboratory. The percentages and counts of B-cell subsets of 33 patients and 10 healthy controls and the immunoglobulin M expression on B-cell subsets of 20 patients and five healthy controls were quantified by flow cytometry. Immunoglobulin levels produced by B cells after stimulation in vitro of 20 patients and five healthy controls were tested by enzyme-linked immunosorbent assay. Redistribution and selective depletion of B-cell subsets in septic shock patients were discovered, and a decrease in immunoglobulin M levels was associated with a reduction in resting memory B-cell counts. These alterations were more pronounced in nonsurvivors compared with survivors. Additionally, receiver operating characteristic curve analysis showed that the data of B-cell subsets had the best predictive value for mortality risk.

Conclusions:

Severe B-cell abnormalities are present in the immunosuppressive phase of septic shock and are associated with prognosis.