bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and functional analysis of variants in ISL1 promoter from patients with isolated atrial septal defects
Authors: Yin Xiu‐Yun , Chen Huan‐Xin , Chen Zhuo , Han Jun , Yang Qin , He Guo‐Wei
Published: May 2022

Objective

Atrial septal defect (ASD) is one of the most common types of congenital heart disease (CHD). As a transcription factor, ISL1 is expressed specifically in second heart field during embryogenesis and plays a crucial role in heart development and cardiovascular morphogenesis. Studies have shown association of ISL1 multiple variants with diverse type of CHD including ASD, whereas the variants of ISL1 gene promoter region in ASD patients have not been reported. We hypothesized that variants in ISL1 promoter may be involved in the formation of ASD.

Methods

Blood samples from 593 subjects, including 300 patients with isolated ASD and 293 healthy controls were studied. Total DNA from all subjects was extracted and the PCR combined with Sanger sequencing was used to identify ISL1 gene promoter variants. Functional analysis of DNA sequence variants was performed using a dual luciferase reporter assay. Electrophoretic mobility shift assay (EMSA) was selected to examine the influence of the variants in ISL1 gene promoter on binding of transcription factors. All possible binding sites of transcription factor influenced by the identified variants were predicted by the PROMO database.

Results

Four variants in ISL1 gene promoter were found only in patients with ASD by sequencing. Three of the 4 variations [g.4923 G>C (rs541081886), g.5079 A>G (rs1371835943), g.5309 G>A (rs116222082)] significantly decreased the transcriptional activities compared with the wild‐type ISL1 gene promoter (P < 0.05). The EMSA revealed that these variants [g.4923 G>C (rs541081886), g.5079 A>G (rs1371835943), g.5309 G>A (rs116222082)] in ISL1 promoter affected the number and affinity of binding sites of transcription factors. Further analysis with the online PROMO database demonstrated that a cluster of putative binding sites for transcription factors may be altered by these variants.

Conclusion

These sequence variants identified from the promoter region of ISL1 gene in ASD patients are likely involved in the development of ASD by affecting the transcriptional activity and altering ISL1 levels. Therefore, these findings may provide new insights into the molecular etiology and potential therapeutic strategy of ASD.
Communicating Science at 24 fps
Authors: Khadria Ambalika S. , Rajan Ranga
Published: May 2022

Video storytelling is a powerful way to share science with the research community and general public. Films can illustrate complex scientific concepts and democratize access to high quality science. In this work, we describe three categories of films created by us in collaboration with students, postdocs and principal investigators. First, behind‐the‐scenes videos of in‐progress experiments, shot in the lab and annotated with relevant context, provide global audiences with a candid view of the scientific process and personas. Second, promotional films created for well‐defined R&D stakeholder groups such as academic departments and graduate programs serve to highlight research philosophies, core values and technical strengths. Third, showcase videos on state‐of‐the‐art instrumentation facilities enable leadership and potential donors to appreciate shared‐resource models that exemplify collaboration and facilitate ambitious research programs. These videos address important goals for the research community such as faculty/student recruiting, fundraising and science advocacy. The biomedical research community would be well‐served in its overarching goals by embracing films as essential components of an increasingly complex science communication ecosystem.

At KathASK Films, we combine more than 25 years of life sciences research experience with a passion for creative art and storytelling to produce impactful science videos. Our interdisciplinary background in basic research and sensitivity towards under‐represented attributes such as gender bias, racial disparity and mental health stigma allows us to weave the science and its underlying human element into engaging stories.
Time‐of‐Day Influence on Central Nervous System Autoimmunity
Authors: Barnoud Coline , Wang Chen , Ince Louise M. , Scheiermann Christoph
Published: May 2022

Circadian rhythms are a characteristic of most living organisms and describe biological processes experiencing an oscillation of about 24h. These rhythms regulate multiple physiological systems, including innate and adaptive immune responses. Circadian disruption is an important risk factor in neuroinflammation such as multiple sclerosis (MS), a T‐cell mediated autoimmune disease affecting around 2.8 million people worldwide. In the central nervous system (CNS), autoimmune T cells attack oligodendrocytes leading to progressive demyelination and neurological impairment. Studies performed by our lab demonstrate that the circadian clock plays an important role in the severity of experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis. However, the mechanisms responsible for this time‐of‐day difference are unknown. We investigated time‐of‐day differences in adhesion molecule expression and immune cell infiltration to the CNS that could explain differences in disease phenotype. Our results indicate a time‐of‐day‐dependent infiltration of cells into spinal cords during EAE development that correlates with expression of the adhesion molecules VCAM‐1 and ICAM‐1 in the CNS vasculature. Targeting leukocyte‐endothelial cell interactions in a time‐of‐day dependent manner with an anti‐VLA4 antibody that blocks VLA‐4/VCAM‐1 engagement reduces leukocyte infiltration and disease severity. This study shows the importance of circadian oscillations in adhesion molecule expression and cell infiltration in the CNS, and identifies an axis to target for chronotherapy purposes in MS patients.
Central Adenosine A1 Receptors Arbitrate the Nicotine Counteraction of Cardiovascular and Autonomic Dysfunctions in Septic Rats
Authors: El‐Naggar Amany E. , Helmy Mai M. , El‐Gowilly Sahar M. , El‐Mas Mahmoud M.
Published: May 2022

The cholinergic anti‐inflammatory pathway plays a crucial role in improving end‐organ damage provoked by inflammatory disorders. Given the interplay between cholinergic and adenosinergic neuronal circuits in the central nervous system, we tested the hypothesis that central adenosine A1 receptors (A1Rs) modulate the offsetting action of nicotine on cardiovascular and inflammatory insults induced by sepsis. Pharmacologic agonistic and antagonistic studies were conducted in conscious rats, pre‐instrumented with femoral vessel and intracisternal (i.c.) catheters. Sepsis was induced by cecal ligation and puncture (CLP) 24 hr before cardiovascular measurements. Nicotine (25 or 100 µg/kg i.v.) dose‐dependently reversed septic manifestations of hypotension and concomitant reductions in time and spectral measures of heart rate variability (HRV) and cardiac sympathovagal balance (low‐frequency/high‐frequency ratio). Like nicotine, the i.c. administration of N(6)‐cyclopentyladenosine (CPA, AR1 agonist) to CLP rats caused significant increases in indices of HRV and sympathovagal balance and greater surges in these parameters were noted upon simultaneous administration of CPA plus nicotine. We also show that the favorable influences of nicotine on blood pressure and HRV in septic rats were significantly inhibited after central blockade of A1Rs by i.c. 8‐Cyclopentyl‐1,3‐dipropylxanthine (DPCPX). Immunohistochemically, the alleviation by nicotine of the upregulated myocardial expression of NFκB in septic rats disappeared upon prior exposure to DPCPX. The data suggest that central adenosine A1Rs act tonically to facilitate the ameliorative effect of nicotine against the inflammatory response of sepsis and interrelated cardiomyopathic and neuropathic hitches.
Anatomical Variants of the Intercostobrachial Nerve and their Clinical Implications in Axillary Dissection
Authors: Olivencia‐Delgado Mariana N. , Jusino‐Álamo Javier , De Miranda‐Sánchez Emanuel , Jiménez‐Dietsch Sofía , Quiñones‐Rodríguez Jailenne
Published: May 2022

Currently, there are multiple treatment options for breast cancer including chemo‐ and radiotherapy. When these treatment options do not work, a radical mastectomy may be considered. This procedure includes an axillary lymph node dissection. During this type of dissection, the intercostobrachial nerve (ICBN) is commonly encountered. Originating from the T2 spinal level, the ICBN is a lateral cutaneous branch of the second intercostal nerve. After piercing the intercostal and the serratus anterior muscles, it traverses the axilla to the medial side of the arm and joins the medial cutaneous nerve of the arm. Here, we report ICBN variations on two distinct cadavers encountered during dissection of the axilla at the Department of Anatomy of the Universidad Central del Caribe School of Medicine. One of the ICBN has three branches on the left upper extremity, while another cadaver has two branches on the right upper extremity. It is important to be aware of these variations while performing an axillary lymph node dissection for breast cancer or any other procedure that involves the axillary region (e.g., regional nerve blocks). Lesions of the ICBN have been associated to post‐operative pain, paresthesia, and loss of upper extremity sensation in the dermatome supplied by this nerve. Therefore, preservation of the ICBN is an important aspect of axillary region procedures.
Loss of ciliary adenylyl cyclase 3 in the ventromedial hypothalamus resulted in a sex‐dependent obese phenotype
Authors: Deng Guorui , Saito Kenji , Singh Uday , Deng Yue , Dickey Jacob , Jiang Jingwei , Cui Huxing
Published: May 2022

Obesity, a metabolic disorder characterized by an excessive accumulation of body fat, continues to be a major economic and health burden to the global populace, as it is the most significant risk factor for heart disease, diabetes, and hypertension. Genome‐wide association studies have identified several genes involved in obesity development, including ADCY3, a gene encoding for the enzyme adenylyl cyclase 3 (AC3). AC3 is a critical mediator of the cyclic adenosine monophosphate (cAMP) signaling pathway and is enriched in the primary cilium, a solitary immotile organelle protruding from most mammalian cells, including neurons. The primary cilia act as the cellular antenna to sense and relay changes in the extracellular microenvironment and mounting evidence support a role for the primary cilia as a critical regulator of the energy homeostasis. Patients carrying loss‐of‐function ADCY3variants are obese and are at increased risk of developing type II diabetes mellitus (T2DM). Consistently, whole‐body Adcy3knockout mice also develop obesity; however, the underlying mechanisms by which AC3 affects metabolic homeostasis remain unclear. Here, we examined the impact of AC3 loss within the ventromedial nucleus of hypothalamus (VMH), a brain region critical for coordinated control of energy and glucose homeostasis, on energy metabolism. We generated mice with VMH‐specific Adcy3loss (Adcy3SF‐1KO mice) by crossing SF‐1 Cre+ mice with Adcy3F/F mice and subjected them to metabolic phenotyping to assess energy balance and glucose homeostasis. We observed a sexually dimorphic effect on body weight under standard chow diet; male Adcy3SF‐1KO mice displayed normal body weight gain comparable to control littermates, whereas female Adcy3SF‐1KO mice gained a significant more weight compared to control littermates (Con 24.34g v KO 30.61g, p < 0.0001). Strikingly, however, both sexes of Adcy3SF‐1KOmice exhibited improved glucose tolerance (AUC: M Con 17,826 v KO 15,165, p = 0.02; F Con 15,263 v KO 13,238, p = 0.03) without affecting fed and overnight fasting blood glucose levels. Taken together, these results underscore the importance of AC3‐mediated ciliary cAMP signaling in VMH neurons for the regulation of energy balance and glucose homeostasis in a sex‐dependent manner.
Pathophysiological role of NNAT in ER+ breast cancer
Authors: Semenikhina Marharyta , Plasterer Cody , Tsaih Shirng‐Wern , Flister Michael , Palygin Oleg
Published: May 2022

Neuronatin (NNAT) was recently identified as a novel mediator of estrogen receptor‐positive (ER+) breast cancer cell proliferation and migration, which correlated with decreased tumorigenic potential and prolonged patient survival. However, despite these observations, the molecular and pathophysiological mechanism(s) of NNAT in ER+ breast cancer remain unclear. Based on high protein homology with phospholamban, we hypothesized that NNAT mediates homeostasis of intracellular calcium [Ca2+]i) levels and endoplasmic reticulum (EndoR) function, which is frequently disrupted in ER+ breast cancer and other malignancies.

To evaluate the role of NNAT on [Ca2+]i homeostasis, we used intracellular labeling and subcellular confocal imaging of multiple breast cancer models (ER+ breast cancer cell lines T47D and ZR75, MCF10A normal mammary epithelial cells, and MDA‐MB‐231 triple‐negative breast cancer cell lines originally obtained from ATCC (American Type Culture Collection) to better understand the role of NNAT in Ca2+‐mediated cellular functions.

Our data indicate that NNAT localizes predominantly to EndoR and lysosome, and genetic manipulation of NNAT levels demonstrated that NNAT modulates [Ca2+]i influx and maintaining Ca2+homeostasis. The application of thapsigargin and consecutive inhibition of SERCA in ER+ breast cancer cells overexpressed NNAT show a significant elevation in the amount of Ca2+ in EndoR (184 ± 5 %, p<0.001 comparing to GFP). At the same time, overexpression of NNAT increased basal cytosolic Ca2+ concentration up to 50% (p<0.001 comparing to GFP). Pharmacological inhibition of calcium channels revealed that NNAT regulates [Ca2+]i levels in breast cancer cells through the interaction with ORAI1 but not the TRPC3 signaling cascade. Furthermore, NNAT transcriptionally regulated by NRF1, PPARα, and PPARγ and is strongly upregulated by oxidative stress via the ROS and PPAR signaling cascades.

Collectively, these data suggest that NNAT regulates Ca2+homeostasis in response to oxidative stress, thus providing a molecular link between the longstanding observation that accumulating ROS and altered Ca2+signaling are key oncogenic drivers of cancer.
Optimization and Validation of Nanopore Based Sequencing Method for Molecular Testing of CNS Tumours
Authors: Mimosa Mashiat L. , Al‐ameri Wafa , Nakhla Michael , Simpson Jared T. , Boissinot Karel , Munoz David G. , Das Sunit , Fattouh Ramzi , Saleeb Rola
Published: May 2022

Background

The World Health Organization (WHO) introduced molecular identifiers for the diagnosis and prognosis of CNS tumors including the mutational status of isocitrate dehydrogenase or IDHgenes in glial tumors. Currently used immunohistochemistry (IHC) is not capable of detecting the non‐canonical mutations, and sequencing is often required as a follow‐up. Current next‐generation sequencing (NGS) technologies used in tumor molecular marker detection introduce key challenges including high capital cost, complex infrastructure requirements, and long turnaround times. These challenges considerably limit the ability to perform NGS testing in many pathology laboratories. In this study, we aimed to use third generation nanopore sequencing technology to resolve these limitations. The Oxford Nanopore MinION, a pocket‐sized nanopore sequencing device, has minimal capital costs and infrastructural requirements, and shorter turnaround times. However, the nanopore technology has not been validated in clinical practice and has not been optimized on formalin‐fixed paraffin‐embedded (FFPE) tissue.

Methods

DNA extraction of selective tumor areas was performed from the corresponding FFPE tissue blocks from a cohort of gliomas with confirmed IDH1and IDH2gene statuses (n=65). A PCR amplicon‐based approach was used to amplify hot spots of the IDH1and IDH2genes, starting with 30ng DNA material. The amplicon libraries were sequenced for 2 hours in multiplex on the MinION device and IDHSNPs were called with the Nanopolish software. ASIP Abstract Mashiat Mimosa

Results

26 IDH mutant samples were identified: 21 IDH1R132H, 2 IDH1R132G, 2 IDH2R172G, and 1 IDH2D177H. All cases showed concordant IDHmutational status when compared to the reference methods (IHC or NGS) and both analytical sensitivity and specificity were 100%. Precision analysis of variant allele frequency (VAF) showed the coefficient of variation was less than 5% (both inter and intra runs), and the limit of detection for VAF was 3%. The range of read depth obtained was 882X to 43,000x with an average of 20,000x. This assay revealed a $50‐$100 material cost per sample, and the time taken from extracted nucleic acid to final result generation was 1‐2 business days.

Conclusion

This project is the first to optimize and validate an approach to detect SNP mutations in FFPE samples using nanopore technology. It has demonstrated the feasibility and efficacy of the nanopore amplicon sequencing method in cancer FFPE tissue with excellent test performance characteristics, significantly shorter turnaround times at considerably lower costs and without any infrastructural needs. Thus, it can be used to circumvent challenges to current NGS testing platforms and can be the milestone that would make cancer NGS testing available for every laboratory.
Safety and Efficacy of a Novel Dietary Pyrroloquinoline Quinone Disodium Salt on Cognitive Functions in Healthy Volunteers: A Clinical Investigation
Authors: Shiojima Yoshiaki , Takahashi Megumi , Takahashi Ryohei , Moriyama Hiroyoshi , Bagchi Debasis , Bagchi Manashi , Akanuma Masanori
Published: May 2022

Cognitive dysfunctions are increasing alarmingly around the world, and researchers are exploring preventive measures for improving brain performance. Pyrroloquinoline quinone (PQQ), a naturally occurring coenzyme in foods, exhibits potent antioxidant activity, and improves diverse functions which includes mitochondrial activation, growth, repair, protection of nerve cells by increased expression of nerve growth factor (NGF) and NGF receptors; and suppression of fibril formation and aggregation of amyloid β. A randomized, double‐blind, placebo‐controlled, parallel‐group clinical investigation (RCT) was conducted over a period of 12 weeks of supplementation of PQQ disodium salt (mnemoPQQ®; p.o.; 21.5 mg/day) in 64 healthy Japanese male and female volunteers (age 40 ‐ <80 Y) to determine the safety and efficacy on improved cognitive function and performance. In another independent study, mnemoPQQ® exhibited broad spectrum safety in a battery of toxicological assessments. No mutagenic potentials were observed. The efficacy of mnemoPQQ® on cognitive performance (memory, attention, judgment, and cognitive flexibility) was examined using Cognitrax as the primary outcome (endpoint), and forgetfulness questionnaire (DECO: Deterioration Cognitive Observee) and Mini‐Mental State Examination‐Japanese (MMSE‐J) as the secondary outcomes (endpoints). A total of 58 subjects (Placebo group = 31; Age = 70.91 ± 3.06 Y, and mnemoPQQ® group = 27; Age = 72.10 ± 3.77 Y) completed the study over a period of 12 weeks. Significant improvements were observed on the Cognitrax’s cognitive function domain score on “composite memory”, “verbal memory”, “reaction time”, “complex attention”, “cognitive flexibility”, “executive function”, and “motor speed” in the mnemoPQQ® group as compared to the placebo group. The DECO index and the MMSE‐J score were also significantly improved in the mnemoPQQ® group. No adverse events were reported. This investigation demonstrates that supplementation of mnemoPQQ® is safe and efficacious in improving memory, attention, judgment, and cognitive function, in middle‐aged to elderly population.
Cerebrovascular Carbon Dioxide Reactivity with Hyperoxia and Hypoxia in Humans with Treated Hypertension
Authors: Fan Jui‐Lin , Sayegh Ana L. C. , Dawes Mathew , Paton Julian F. R. , Fisher James P.
Published: May 2022

Hypertension increases the risk of cerebrovascular diseases such as stroke and dementia, but the underlying mechanisms are currently unclear. While chronic hypertension can lead to anatomical and functional adaptations within the cerebral circulation, its effect on cerebral blood flow (CBF) control remains poorly understood. We tested the hypothesis that the CBF responses to CO2 with either high or low oxygen levels are impaired in treated hypertensive patients compared to normotensive aged‐matched controls.

In 14 treated hypertensive patients (age: 68±5 years, mean±SD) and 8 normotensive age‐matched controls (age: 66±6 years), we assessed cardiorespiratory and cerebrovascular variables at rest, during voluntary hyperventilation (partial pressure of end‐tidal CO2 [PETCO2] ~25 mmHg), and hyperoxic and hypoxic rebreathing. Using transcranial Doppler ultrasound and finger photoplethysmography, we measured beat‐to‐beat middle cerebral artery blood velocity (MCAv: index of CBF) and BP, respectively, while PETCO2 was assessed using a gas analyzer. Linear regression was used to determine the MCAv‐CO2 slope (i.e., cerebrovascular CO2 reactivity).

At rest, mean BP was higher in the hypertensive patients (108±9 mmHg) compared to normotensive controls (93±10 mmHg, p=0.001), while MCAv (46.8±11.4 vs. 53.3±10.4 cm/s) and PETCO2 (41±4 vs. 42±3 mmHg) were not different between groups (p<0.05). Hyperventilation‐induced hypocapnia elicited similar reductions in MCAv in the hypertensive and normotensive groups (Δ14.8±4.7 vs. Δ16.5±5.7 cm/s, p=0.484). Similarly, the MCAv‐CO2slope was not different between the hypertensive and normotensive groups during either hyperoxic rebreathing (1.53±1.05 vs. 1.98±0.56 cm/s/mmHg, p=0.320) or hypoxic rebreathing (1.94±1.04 vs. 2.21±1.13 cm/s/mmHg, p=0.815).

These preliminary findings indicate that in treated hypertensive patients, the MCAv responses to hypocapnia, and hypercapnia administrated with either high or low oxygen levels, do not differ from that observed in age‐matched normotensive controls.
PDE4D inhibitor: a novel weapon against mild cognitive impairment and Alzheimer’s disease?
Authors: Xu Ying
Published: May 2022

A classical hallmark of Alzheimer’s disease (AD) is the accumulation of amyloid‐β (Aβ), which correlates significantly with progressive cognitive and learning and memory impairment. BPN14770, an allosteric inhibitor of PDE4D, has been proposed to improve memory deficits induced by Aβ oligomers (AβOs) and provide overall AD pathology improvement. However, the causal relationship between PDE4D‐related cAMP signaling and the prevalence of AβOs‐induced cognitive deficits has not been investigated. The present study examined whether the memory and cognitive enhancing effects of BPN14770 in humanized PDE4D‐APP/PS1 (hPDE4D/Tg‐AD) mice, which are engineered to express a primate‐specific N‐terminal region of PDE4D and crossed with transgenic APP/PS1 mice, were directly involved in reduced cAMP signaling dependent amyloid pathway. The results suggested that the PDE4D inhibitor BPN14770 significantly improved cognitive index in novel object recognition task, and memory acquisition and retrieval in Morris water maze test in h hPDE4D/Tg‐AD mice. Pretreatment with protein kinase A (PKA) inhibitor H89 completely blocked this BPN14770‐induced memory and cognitive enhancement, suggesting the key role of PDE4D dependent cAMP signaling in cognitive processes. Furthermore, the hPDE4D/Tg‐AD mice showed increases in Aβ plaque and soluble/insoluble Aβ concentration both in the cortex and hippocampus. While treatment of BPN14770 for two weeks (via gavage) prevented these pathological changes, which can be blocked by H89. The subsequent studies suggested that BPN14770 affected the synthesis and degradation of Aβ as evidenced by decreases in amyloidgenic and non‐amyloidgenic pathways such as BACE‐1, sAPPβ, ADAM10 and sAPPα protein expression, and increases in Aβ degrading enzymes including IDE and NEP levels, and downstream neuroprotective proteins such as pCREB/CREB and BDNF expression in hAPP/PS1‐PDE4D mice. Further chromatin co‐immunoprecipitation assays showed cAMP‐dependent CREB directly increased ADAM10 and IDE expression, and inhibited BACE‐1 expression through NF‐κB, resulting in reduction of Aβ production and increase in Aβ degradation. These findings reveal a causal relationship between PDE4D signaling and the progress of AD and demonstrate the clinical potential of BPN14770 in the treatment of AD.
Synthetic Cannabidiol Reduced Body Mass and Visceral Adiposity but Not Blood Pressure in Rats With Advanced Metabolic Syndrome
Authors: Wilson Jessica N. , Mendez Dora , Mascal Mark , Fitzgerald Robert , Ortiz Rudy M.
Published: May 2022

Very little is known about the effects of CBD on age‐associated cardiovascular and metabolic dysfunction. The prevalence of metabolic syndrome (MetS) is 54.9% among people ≥60 years of age, which increases the risk of the development of associated pathologies like cardiovascular disease and type II diabetes. Additionally, synthetic CBD alternatives offer a chemically pure product, which is free of plant‐associated contaminants. To assess the effects of the synthetic CBD, H4CBD, on advanced metabolic dysfunction, a cohort of 41‐week‐old lean strain Long‐Evans Tokushima Otsuka (LETO) rats (n=8) and MetS strain Otsuka Long‐Evans Tokushima Fatty (OLETF) rats were administered sesame oil vehicle (n=8) or 200 mg H4CBD/kg (OLETF n=8) by oral gavage for 4 weeks. Animals were fed standard chow ad libitum and monitored for systolic blood pressure (SBP), activity, body mass (BM), food intake and urinary excretion. After 4 weeks, H4CBD tended to increase activity scores in OLETF (p=0.099) and reduced BM to control levels within the first week. Retroperitoneal and epidydimal fat masses were reduced 25% compared to untreated OLETF. However, the H4CBD‐induced loss of visceral adiposity was not associated with reduced arterial blood pressure. Clinical indicator for muscle wasting, urinary 3‐methylhistidine (3MH), ratioed to urinary creatinine (Cr) (3MH/Cr), suggested the primary mass lost was adipose. An oral glucose tolerance test (oGTT) was performed after 4 weeks of treatment. Glucose response was reduced (AUCglucose) 29% (p<0.001) by H4CBD treatment compared to control and reduced insulin resistance index (IRI) by 25% (p<0.05). However, H4CBD‐induced improvements to glucose tolerance did not associate with reduced basal blood glucose or plasma insulin, despite compensatory increases in insulin receptor (IR) protein expression (54%; p<0.05) and membrane‐bound glucose transporter protein 4 (GLUT4) protein expression (41.5%). The results suggest that a chronic, high dose of CBD may improve glucose tolerance, independent of insulin signaling, without caloric restriction via increased activity. H4CBD reduced plasma triglycerides by 45% (p<0.05) but not plasma NEFA suggesting that de novo TG synthesis was reduced. The results suggest that a chronic, high dose of CBD may drive fat loss via increased activity and reduced TG synthesis, but the mass loss was not sufficient to correct the associated hypertension or impaired substrate metabolism common in MetS for this duration of treatment and advanced stage of dysregulated metabolism.
Middle‐Aged and Older Adults with Greater White Matter Hyperintensity Burden Display Attenuated Blood Pressure and Cerebral Blood Velocity Responses During Orthostatic Tilt
Authors: Iftikhar Rehan , Pearson Andrew , Miller Kathleen , Corkery Adam , Eisenmann Nicole , Howery Anna , Chin Nathaniel , Johnson Sterling , Barnes Jill
Published: May 2022

Cerebral autoregulation (CA) is the process by which the brain maintains adequate blood flow despite sudden changes to perfusion pressure. White matter hyperintensities (WMH) are associated with increased risk of cerebrovascular disease and dementia. This study investigated whether dynamic CA is reduced in cognitively unimpaired middle‐aged and older adults. Additionally, we evaluated dynamic CA and cerebrovascular responses at rest and in response to orthostatic tilt. We hypothesized that CA would be impaired during orthostatic tilt and that diminished blood pressure and cerebral artery blood velocity changes would be associated with greater WMH fraction. We studied 33 middle‐aged and older adults (age = 63 ± 4 y). Intracranial volume (ICV) and WMH lesion volume were measured using a T1 and FLAIR scan, respectively, on a 3T MRI scanner. WMH fraction was calculated as the cube root of (WMH lesion volume/ICV) *100. CA was measured using middle cerebral artery blood velocity (MCAv) from transcranial Doppler ultrasound and beat‐to‐beat mean arterial pressure (MAP) from a finger cuff. Five minutes of data were collected at rest and during 60º orthostatic tilt. Transfer function analysis was used to quantify dynamic CA and reported as gain and phase at low frequency (LF) (0.07‐0.20 Hz). MCAv, MAP and cerebrovascular resistance index (CVRi; calculated as MAP/MCAv) were used to quantify the cerebrovascular response to orthostatic tilt. LF gain was greater at baseline compared to 60º tilt (0.52 cm/s/mmHg vs 0.46 cm/s/mmHg, p < 0.05); however, there were no differences in LF phase between baseline and 60º tilt (29.70 degrees vs 29.42 degrees, p = 0.95). During 60º tilt, MAP and MCAv decreased compared to baseline (MAP: 106 mmHg vs 101 mmHg, p = 0.02; MCAv: 59 cm/s vs 53 cm/s, p < .01). CVRi was higher at 60º tilt compared with baseline (1.90 mmHg/cm/s vs 2.02 mmHg/cm/s, p < 0.01). There were significant inverse associations between the percent change from baseline to 60º tilt in MAP (r = ‐0.4, p = 0.03), MCAv (r = ‐0.39, p = 0.03), but not CVRi (r = 0.13, p = 0.50), and WMH fraction. There were no associations at baseline or 60º tilt between gain (Baseline: r = ‐0.18, p = 0.33; 60º: r = ‐0.31, p = 0.09) or phase (Baseline: r = 0.07, p = 0.71; 60º: r = ‐0.29, p = 0.12) and WMH fraction. Our results indicate that dynamic CA is preserved in cognitively unimpaired adults during tilt. However, individuals with diminished MAP and MCAv responses to orthostatic tilt displayed greater WMH fraction. Future studies are needed to investigate the relationship between blood pressure and cerebral blood flow in individuals with cognitive impairment to further understand the role of cerebral blood flow regulation in the development of cognitive decline.
A selenium‐iron axis dictates cancer cell sensitivity to pharmacologic ascorbate
Authors: Jankowski Connor S. , Rabinowitz Josh
Published: May 2022

High‐dose ascorbate (vitamin C) has shown promising anti‐cancer activity. We sought to distinguish the mechanism of cancer cell ascorbate toxicity between two proposed models: hydrogen peroxide (H2O2) generation by ascorbate itself or glutathione depletion by its oxidized form, dehydroascorbate. Using a combination of metabolic and genetic approaches, we show that ascorbate kills cancer cells through a free radical mechanism that is promoted by iron and suppressed by selenium. High‐dose ascorbate’s metabolic effects and cytotoxicity result from hydrogen peroxide independent of dehydroascorbate. Cytotoxicity further depends on iron via a route distinct from canonical ferroptosis, as the hydrogen peroxide‐detoxifying selenoenzyme GPX1 is critical while the ferroptosis‐suppressing GPX4 is dispensable. Selenium‐mediated protection from ascorbate is powered by NADPH from the pentose phosphate pathway. In a mouse model of glioblastoma, dietary selenium deprivation enhances the efficacy of ascorbate as an anti‐cancer agent. These data establish iron and selenium as opposing mediators of high‐dose ascorbate’s pharmacological activity. More generally, they suggest that cancer sensitivity to free‐radical therapies depends on mineral bioavailability.
Uncovering the neural circuits that drive food odor detection in drosophila
Authors: Mostyn Jack
Published: May 2022

The ability to correctly identify food is critical to survival. Smell is a primary tool used by many animals to find food, yet much is still unknown about the process. Our research is aimed at uncovering the neural circuits that drive food odor detection. We used optogenetic manipulations, a variety of behavioral assays, and immunostaining to study food odor detection in fruit flies. Our results indicate that food odor detection involves both learned and innate components; challenging the notion that food odor detection is wholly innate.
Sympathetic Neurocirculatory Responses to Chemoreflex Activation in Young Women and Men
Authors: Sayegh Ana Luiza C. , Fan Jui‐Lin , Dawes Mathew , Paton Julian F. , Fisher James P.
Published: May 2022

Sex‐differences in the ventilatory responses to central and peripheral chemoreflex activation have been extensively examined. Along with the control of respiration, chemoreflexes play a key role in sympathetic regulation. Herein, we determined whether central, peripheral, and combined central and peripheral chemoreflex activation evoke differing sympathetic neurocirculatory responses in young women and men.

In ten women (29±6 years, 23±2kg/m2: mean±SD) and ten men (30±7 years, 25±3kg/m2), we recorded muscle sympathetic nerve activity (MSNA), minute ventilation (V̇E) and mean arterial pressure (MAP) during 5‐min trials of breathing room‐air (eucapnia), isocapnic hypoxia (10% oxygen [O2]; peripheral chemoreflex activation), hypercapnic hyperoxia (7% carbon dioxide [CO2], 50% O2; central chemoreflex activation) and hypercapnic hypoxia (7% CO2, 10% O2). Trials were randomized. Women were studied in the early follicular phase of their menstrual cycle.

During eucapnia and isocapnic hypoxia, no sex‐differences were observed in MSNA, V̇E and MAP (P>0.05). During hypercapnic hyperoxia (∆278±215 vs. ∆158±107%, P=0.05) and hypercapnic hypoxia (∆507±290 vs. ∆262±268%, P=0.05) MSNA responses were augmented in women compared to men, however V̇E responses were attenuated (hypercapnic hyperoxia ∆15.2±6.7 vs. ∆25.6±6.7L/min, P=0.007; hypercapnic hypoxia ∆28.8±9.3 vs. ∆39.9±12.5L/min, P=0.004). MAP responses were similar in women and men (hypercapnic hyperoxia ∆10±2 vs. ∆23±2mmHg, P=0.35; hypercapnic hypoxia ∆16±3 vs. ∆25±3mmHg, P=0.35).

In summary, young women have greater MSNA response to central chemoreflex activation than young men, which explains their augmented sympathetic response to combined central and peripheral chemoreflex activation. Future studies should determine the effect of menopause on these sex‐differences in the sympathetic neurocirculatory responses to chemoreflex activation.
Altered Mitochondrial Dynamics in Hepatocytes Induces Nonalcoholic Fatty Liver Disease Progression
Authors: Miyao Masashi , Kawai Chihiro , Kotani Hirokazu , Tamaki Keiji , Eguchi Satoru
Published: May 2022

Nonalcoholic fatty liver disease (NAFLD) is a hepatic manifestation of metabolic syndrome that affects approximately 25% of adults globally. Although several pharmacological therapies have been reported to improve NAFLD, the therapeutic ranges of the drugs are limited. Mitochondrial dysfunction could be a key factor involved in NAFLD progression. However, the association between mitochondrial dynamics, which involves mitochondrial fission and fusion, and NAFLD development has not been fully characterized to date. Herein, we examined the effects of alterations of mitochondrial dynamics in hepatocytes and liver sinusoidal endothelial cells (LSECs), the most abundant non‐parenchymal cells in the liver, on NAFLD development and hypothesized that rebalancing of mitochondrial dynamics in these cells could attenuate NAFLD progression. Mice were fed a choline‐deficient, L‐amino acid‐defined diet (CDAA) for 1 or 4 weeks to recapitulate human NAFLD development. Malformed mitochondria, such as megamitochondria and mitochondria with deformed cristae, were substantially increased in both, hepatocytes and LSECs, of mice that were fed CDAA diet, and were associated with NAFLD development. In addition, decreased mitochondrial content and increased mitochondrial number, suggesting excessive mitochondrial fission, were induced in hepatocytes, but not in LSECs, after four weeks of CDAA feeding. These unfavorable changes in the mitochondria of hepatocytes, but not of LSECs, were attenuated by mdivi1, a chemical mitochondrial fission inhibitor. Liver inflammation and fibrosis were also attenuated by mdivi1, as assessed by serum biochemical and histochemical analyses. The expression of several inflammatory and fibrogenic genes (Tnfα, Il6, and Col3α1) tended to decrease with mdivi1 treatment. The expression of genes related to mitochondrial biogenesis, fission, fusion, and mitophagy was also significantly reduced by mdivi1 (p<0.05), suggesting rebalanced mitochondrial dynamics. Ultrastructural analyses revealed that endoplasmic reticulum stress and autophagy in hepatocytes were mitigated by mdivi1 administration. Taken together, these findings suggest that excessive mitochondrial fission in hepatocytes induces alterations in mitochondrial dynamics and other organellar injuries, and subsequently, liver inflammation and fibrogenesis. Therefore, rebalancing mitochondrial dynamics in hepatocytes by inhibiting excessive fission is a promising therapeutic strategy for NAFLD.
The Design of an Antagonist that Improves Cardiac Function through Ligand‐Directed Signaling
Authors: Pittala Karthik , Nichols Ruthann , Maynard Benjamin , Leander Megan , Creech Jeff , da Rocha Andre Monteria , Heron Todd
Published: May 2022

The signaling pathway of a peptide that impacts cardiovascular physiology may be targeted to influence cardiac function. Through previous studies, we discovered that human RFRP‐1 (RFamide‐related peptide‐1; MPHSFANLPLRF‐NH2) decreases cardiac contractility. The structure and activity of RFRP‐1 are conserved across phylogeny, suggesting it plays a role in physiology. We determined RFRP‐1 structure‐activity relationship (SAR) in cardiomyocytes and demonstrated that it acts through RFRP‐1R, a G protein‐coupled receptor (GPCR). Based on RFRP‐1 SAR data, we designed LPLAF‐NH2 and found that it acts as a RFRP‐1R antagonist in cardiomyocytes and in animals. The alanyl‐substituted analog diminished the effects of the full‐length peptide and active core, but not at equimolar concentrations; the antagonist was less effective. In order to design a better antagonist, we turned to molecular docking software to investigate ligand‐receptor binding. As a result, we designed an antagonist with increased receptor interactions that improved cardiac function in human cardiomyocytes and consistent with ligand‐directed RFRP‐1R signaling.
An Online Magnetic Resonance Imaging Approach Using Augmented Reality Visualizations for Introducing Students to Their “Silent Teachers”
Authors: Aytac Gunes , Thompson Jesse , Oktay Cemil , Rettenheimer Chris , Labrash Steven , Lee U‐Y , Lozanoff Scott
Published: May 2022

The initial interaction between medical students and their “Silent Teacher” in the dissection lab is an important experience in the process of establishing a professional ethos. However, COVID19 complicated this process and the optimal approach to achieve this introduction remains unclear. The purpose of this study was to develop an online case‐based approach as the basis for donor introduction to medical students based on Magnetic resonance imaging (MRI) reports. MRI scans of 16 body donors were assessed by a radiologist and observations were uploaded to a web‐based DICOM viewer (www.rad3d.com). A list of learning objectives were generated for each donor and radiology reports were constructed based on: chief complaints, medical history, radiology and pathology that were reviewed prior to the class. Within the lab and following COVID19 restriction policy, students read through the radiology reports and viewed the augmented reality (AR) visualizations of the MRI scans using z‐space computers providing additional dissection team interactions. Finally, surface anatomy exercises were undertaken that corresponded to the primary pathology providing a direct interaction with the donor. This process included Sketchfab and other links to underscore anatomical correlations (Figure 1). A survey was conducted to assess student perceptions of the experience. The study design comprised a voluntary survey tool to assess student level of anxiety and whether the online approach was effective. Response scales were either bimodal or Likert‐base (5 scale) and analyzed with Fisher's exact test. Follow up open ended questions were also presented to the students. The response rate was 33/80 (41.25%). Most students (82.7%) felt anxious first, but the anxiety decreased in time (60.7%). They mostly accomplished the tasks (surface exercises 97%; reviewing MRIs 87.9%) and found surface exercises useful in terms of reducing anxiety (90.6%), and initiating team work (100%). A total of 79.3% of the students found MR useful for learning radiology (Table 1). More than half of the students (55.6%) indicated that the first encounter was not as they expected and stated that they thought it would be more intimidating. Several emotions were indicated such as: confused, uncomfortable, somber, excited, grateful, thankful, professional. They all agreed that using a case based approach as an introductory laboratory prior to first dissection is useful. There was one prominent suggestion almost all students stated: video resources prior to the lab. Results suggest that humanizing the donor, implementing clinical reasoning, and initiating teamwork provided a positive first lab experience using online resources. Future work will be aimed at further development of online resources organized around clinical presentation using MR scans.
LC‐MS based urine metabolomic profiling indicate altered gut microbiome and host lipid metabolites in Parkinson’s disease
Authors: Jewell Shannon , Roper Kerry , Woodhouse Helen , Adam Robert , O'Sullivan John , Gordon Richard
Published: May 2022

Evidence indicates that host and gut microbial metabolism are dysregulated prior to the onset of motor symptoms, in the prodromal phase of Parkinson’s disease (PD). There is currently no definitive test for the diagnosis of Parkinson’s disease; this represents an urgent, unmet medical need to enable the effectiveness of current and future therapeutic interventions for PD. The aim of this study was to understand changes in host and microbial metabolism in PD patient biofluids which could reflect a transition from the healthy to diseased state. We performed a comprehensive untargeted metabolomics analysis of urinary lipid metabolites together with altered metabolites associated with the gut microbiota. A total of 64 urine samples consisting of 31 from age matched healthy controls and 33 from PD patients were used in this study. Samples were deproteinized and analysed by reverse phase (RP)/UPLC‐MS/MS methods with positive and negative ion mode electrospray ionization (ESI) and HILIC/UPLC‐MS/MS with negative ion mode ESI. 14 lipid metabolites were significantly altered in PD patients. Five metabolites were associated with bile acid metabolism and nine were from the cholesterol pathway. Changes in secondary bile acid metabolism, particularly secondary bile acids indicative of microbial metabolism changes, were evident in PD patients, with 11 altered metabolites associated with the gut microbiota being significantly altered. Interestingly, several lipid metabolites that were altered in PD patients correlated with specific gut microbiome metabolites. Our results provide new insights into altered lipid metabolism and gut dysbiosis in PD, however, confirmation in a larger cohort would be needed.
Anti‐inflammatory activity of xylopic acid may be attributed to the modulation of Nrf2 and NF‐kB activities
Authors: Osafo Newman , Boakye Yaw D.
Published: May 2022

For a long time, non‐resolving inflammation has been the bane of our existence [1]. Currently, there are varying treatment and management options for inflammatory conditions. However, the search for newer, alternative management options continues unabated. A key focus of our research is the identification of lead compounds and therapeutically viable compounds from plants. One such compound isolated from Xylopia aethiopica is the kaurene diterpene, xylopic acid (XA). The anti‐inflammatory potential of XA has been established from our previous studies [2, 3]. This current work sets out to identify potential molecular targets of xylopic acid in in vitro and cell‐based assays. The selection of these investigated targets was based on the proven bioactivity of xylopic acid as a potential anti‐inflammatory agent and on predictions from the Spider target prediction webtool. Reporter gene assays were used to test for altered NF‐kB and Nrf2 activities in transfected HEK or CHO cells, respectively. XA at 30 µM significantly (p<0.001) inhibited the NF‐kB‐dependent reporter gene expression (Fig. 1) and enhanced activation of Nrf2 in a concentration‐dependent manner when compared to the control (Fig. 2). Our findings suggest that the anti‐inflammatory mechanism of XA entails the inhibitory effect on NF‐kB and an increased activity of Nrf2.
Fibroblast Senescence: A Risk Factor for Remodeling, Inflammation, and Arrhythmias in the Post‐MI Heart
Authors: Sengun Elif , Baggett Brett , Turan Nilufer , Mi Eric , Kun Bianca , Sedivy John , Koren Gideon
Published: May 2022

Objectives

Cellular senescence is a stress response involving permanent replicative arrest. It is accompanied by a complex senescence‐associated secretory phenotype (SASP), which is characterized by the presence of pro‐inflammatory cytokines and chemokines, growth factors, and tissue‐remodeling metalloproteinases. Little is known regarding the role of senescence and the SASP in the context of arrhythmias post myocardial infarction (MI). Here, we characterized the arrhythmogenic cardiac consequences and molecular mechanisms underlying tissue remodeling changes in impaired or accelerated cardiac myofibroblast (CMF) senescence and the pro‐inflammatory SASP during post‐MI wound healing process in genetically engineered mice.

Material‐Methods

Impaired or accelerated CMF‐specific senescence was achieved by using cre‐lox mouse models to delete p53 or MDM2 genes. MI surgery via direct ligation of the left anterior descending artery (LAD) was performed in 4‐month‐old wild‐type (WT), Pstn‐Cre(+/‐) p53(fl/fl), Pstn‐MerCreMer(+/‐) MDM2(fl/fl), and respective littermate control (LMC) mice of both sexes. Changes in cardiac structure and function post‐MI were assessed by echocardiography. Senescence‐associated β‐galactosidase (SA‐βgal) staining was applied to detect and quantify senescent cells in three zones: infarct zone (IZ), infarct border zone (IBZ), and remote zone (RZ). Immunofluorescence staining identified the types of senescent cells. Masson’s trichrome staining was used for fibrosis assessment. RT‐qPCR was performed to quantitate the expression of known senescence genes and canonical SASP factors.

Results

Pstn‐Cre(+/‐) p53(fl/fl) mice displayed significantly decreased CMF senescence two weeks post‐MI in the scar compared to littermate control p53(fl/fl) and WT mice. We noticed a similar trend in the IBZ, but this did not reach statistical significance. Pstn‐MCM(+/‐) MDM2(fl/fl) mice showed significantly increased CMF senescence in the IZ compared littermate control MDM2(fl/fl) mice. We observed a similar trend in the IBZ, but this did not reach statistical significance. In all mice, we observed essentially no senescence in the RZ. Our data did not show any significant difference in fibrosis between any genotype in any zone. We did not observe any overall trend in differential expression in senescence and SASP genes between p53(fl/fl) and Pstn‐Cre(+/‐) p53(fl/fl) mice in any zone. There was no evidence of any sex‐specific differences in aforementioned results.

Conclusions

A significant increase and persistence of CMF senescence likely contributes to pathological tissue remodeling during the post‐MI wound healing process. This is likely due to SASP‐induced pro‐arrhythmogenic chronic inflammation or via direct cell‐cell interactions between senescent CMFs and myocytes.