bluuebunny/crossref_metadata_2025 huggingface dataset:
Salivary melatonin levels and quality of sleep in second year medical students Antenor Orrego Private University of Trujillo ‐ Peru (LB753)
Authors: Fernández Rodríguez Lissett , Bardales Zuta Victor , Hilario Vargas Julio
Published: June 2021

The goals were to determine the salivary melatonin levels and sleep quality in second year students of Human Medicine of the Antenor Orrego Private University. Of the 268 medical students enrolled in the semester 2013 ‐ I, 40 students were selected by simple random method and met the inclusion criteria. Saliva samples were taken at the beginning, middle and end of the academic semester on Wednesday and Thursday at home at 23:30 and 24 hours. Melatonin levels were determined by ELISA and the Pittsburgh Sleep Quality Index (PSQI) was applied to students. A total of 40 individuals were included in the study: 27 women, 13 men with mean age of 18.45 years (range 16 ‐ 24). Melatonin levels at the start ‐ half 23:30 to 24 hours (p <0.05), start ‐ end 23:30 (p <0.05) and mid ‐ end (p <0.01). The PSQI presented a Cronbach's alpha of 0.553 and averages were 5.9 at the beginning of the semester, middle 7.175 and end 7.475 (p <0.05). Melatonin levels decreased at the beginning and end of the academic semester and according to the PSQI, medical students have poor sleep quality.

Grant Funding Source: Antenor Orrego Private University
Test‐retest reliability of eucapnic voluntary hyperpnea test performance and pre‐post spirometry in elite swimmers (LB783)
Authors: Kennedy Michael , Steinback Craig , Skow Rachel , Parent Eric
Published: June 2021

Test‐retest reliability of eucapnic voluntary hyperpnea test performance and pre‐post spirometry in elite swimmers Michael D. Kennedy1, Craig D. Steinback1, Rachel Skow1, Eric C. Parent2. 1Faculty of Physical Education and Recreation, 2Department of Physical Therapy, University of Alberta, Edmonton, Alberta, Canada.

Exercise induced bronchoconstriction (EIB) is common in “high ventilation” athletes and a 6 min Eucapnic Voluntary Hyperpnea (EVH) airway provocation (30 breaths/min; Tidal Volume (TV) of 85% FEV1; 100% dry medical air) is the standard EIB screen. Sensitivity of EVH is well documented; however, test‐retest reliability of spirometry measurements and EVH performance has not been determined.

Swimmers (n=11; 5/6 m/f) with 蠅13 yrs of competitive experience completed spirometry (FEV1) before and 45 sec, 5, 10, 20 min after an EVH test on 2 consecutive days.

Overall pre‐test FEV1 was decreased on Day 2 (Day 1 4.6±0.8 vs. Day 2 4.5±0.7L, p = 0.04) and this pre‐test FEV1 decrease occurred in 9/11 swimmers (range ‐ 370 (6.8%) to ‐ 30ml (0.6%); with excellent agreement in pre‐test FEV1 between days (ICC 0.97 (0.90 ‐ 0.99, 95% confidence interval). Swimmers achieved 85±9% and 87±9% of target TV on Day 1 and 2 respectively (p = 0.45; ICC 0.57 (0.00 – 0.86)) resulting in a total ventilation of 687 vs 684L (p = 0.89, ICC 0.89 (0.65 – 0.97)) equating to 82±8% and 84±9% of predicted total volume (ICC 0.54 (0.00 – 0.85) between Day 1 and Day 2. Day 1 and 2 post‐test FEV1 were not different at any time point however 4 swimmers were positive for EIB (> 15% fall in pre‐post FEV1) and these swimmers post‐test FEV1 was 10.6 to 1.9% less on Day 2. Post‐test ICC for FEV1 were 0.97 (0.91‐0.99), 0.97 (0.89‐0.99), 0.97 (0.88‐0.99) and 0.94 (0.81‐0.98) at 45s, 5min, 10min, and 20 min respectively.

Overall, strong test‐retest reliability was found for FEV1 and total ventilation; despite a significant decrease in pre‐test FEV1 on Day 2. Post‐test FEV1 is likely more reliable early post‐test (45s, 5min, 10min) than at 20min post‐test.
Adenosine receptor blockade impairs the ability of rat pups to autoresuscitate from primary apnea during repeated exposure to hypoxia (LB790)
Authors: Lun Rongzhi , Zhang Chunfen , Fewell James
Published: June 2021

Autoresuscitation (AR) failure from hypoxic‐induced apnea by gasping has been suggested to play a role in sudden infant death. Little is known, however, about the factors that influence gasping and its ability to effect autoresuscitation in the newborn. The present experiments were carried out on 18 5‐6 day‐old rat pups to investigate the role of adenosine acting via A1 receptors in mediating AR from hypoxic‐induced apnea by gasping. DPCPX (20 mg/kg; a selective adenosine A1‐receptor antagonist) attenuated the heart rate response to hypoxia and decreased the number of hypoxic episodes tolerated before AR failure (DPCPX 10±2 (n=9) versus vehicle 17±5 (n=9); p<0.001). Following vehicle, death was preceded most often by AV dissociation whereas following DPCPX death was preceded most often by an isoelectric electrocardiogram. Thus, our data provide evidence that adenosine acting via A1 receptors plays an important role in promoting AR from hypoxic‐induced apnea by gasping.

Grant Funding Source: CIHR
NF‐κB activation in hindlimb muscles from adult and old mice at rest and following contractile activity (LB814)
Authors: Pollock Natalie , Vasilaki Aphrodite , Lightfoot Adam , Jackson Malcolm , McArdle Anne
Published: June 2021

Skeletal muscle adapts following contractions by increases in reactive oxygen species (ROS) leading to the detection of activation of redox‐responsive transcription factors (TFs) such as nuclear factor kappa B (NF‐κB) by electrophoresis mobility shift assay (EMSA). This TF activation is reduced in muscles from old mice and humans1,2. Additional studies from our laboratory have demonstrated that NF‐κB binding activity of quiescent muscles from old wild‐type (WT) mice was significantly increased compared with adult WT mice1. However, the significance of these findings to changes in transcriptional processes is unclear due to the ex‐vivo nature of EMSA analyses. This study used NF‐κB1 luciferase reporter mice to determine the transcriptional effect of this NF‐κB activation in hindlimb muscles from adult and old mice at rest or following an isometric contraction protocol 1 using immunofluorescence antibody staining of luciferase in transverse frozen sections of tibialis anterior (TA) muscles. Muscles were also examined for evidence of changes in the expression of cytokines known to be under the transcriptional control of NF‐κB. Luciferase staining was observed in all muscle sections with a patchwork distribution between fibres, showing as more intense overall staining in the quiescent muscles from old compared with adult mice. Immediately following the contraction protocol, the luciferase staining was increased in muscles from adult NF‐κB1 mice. In contrast, no increase was seen in the muscles from old mice following contractions compared with quiescent muscle from old mice. The chronic activation of NF‐κB was associated with increased muscle mRNA levels of a number of cytokines. These data confirm previous EMSA observations and suggest that the chronic activation of NF‐κB in muscle from old mice is associated with the generation of a pro‐inflammatory environment and this may restrict the ability in muscles of old mice to further adapt following contractile activity.

Grant Funding Source: Supported by BBSRC and NIA/NIH (AG‐020591)
TWEAK causes cell death in cultured primary myotubes (LB816)
Authors: Ogura Yuji , Kumar Ashok
Published: June 2021

Tumor necrosis factor (TNF)‐like weak inducer of apoptosis (TWEAK) is a proinflammatory cytokine belonging to TNF super family. TWEAK produces a variety of cellular responses through binding to fibroblast growth factor inducible 14 (Fn14), a member of TNF receptor superfamily. Although Fn14 lacks a death domain, TWEAK has been found to induce apoptosis in some cell types by perturbing the activity of certain pathway such as TNFα‐TNF receptor signaling. TWEAK is also known to regulate proliferation and differentiation of myogenic cells. We have previously reported that TWEAK‐Fn14 system causes muscle atrophy both in vitro and in vivo. Moreover, it has been reported that TWEAK mediates muscle atrophy in disuse conditions such as denervation. However, it remains unknown whether TWEAK can affect the viability of muscle cells. To address this issue, we have studied the effects of recombinant TWEAK protein on survival of cultured mouse primary myotubes. Our results demonstrate that TWEAK reduces myotube viability in a dose‐dependent manner evident by increased levels of lactate dehydrogenase (LDH) in culture supernatants. TWEAK also increased the number of the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)‐positive nuclei in cultured myotubes. Furthermore, we have found that the levels of cleaved poly ADP ribose polymerase (PARP) are increased upon treatment with TWEAK for 48 and 72h. These results provide initial evidence that in addition to causing atrophy, TWEAK can also diminish skeletal muscle mass by affecting the survival of myofibers in different catabolic conditions.

Grant Funding Source: Supported by NIH RO1AG029623
Transcriptional regulation of dual‐specificity phosphatase 4 (Dusp4) by muscle‐specific RING finger 1 (MuRF1) (LB822)
Authors: Haddock Ashley , Waddell David
Published: June 2021

Skeletal muscle wasting is a consequence of numerous physiological conditions, including denervation, corticosteroid treatment, and aging. The E3 ubiquitin ligase, MuRF1, is induced under most atrophy conditions and is believed to play a key role in protein degradation in atrophying muscle. However, the preliminary data described in this study provides new evidence that MuRF1 may also act as a transcriptional modulator of atrophy‐induced gene activity, including the regulation of Dusp4 expression. In order to characterize the transcriptional regulation of Dusp4, reporter gene constructs containing fragments of the proximal promoter region of this gene were developed, transfected into C2C12 cells with or without a MuRF1 expression plasmid and analyzed for differences in reporter gene activity.

The Dusp4 reporters showed repressed activity in cells ectopically expressing MuRF1 compared to cells that did not overexpress MuRF1. Furthermore, ectopic expression of the myogenic regulatory factors (MRFs), MyoD1 and myogenin, also caused repression of the Dusp4 reporter constructs, while co‐overexpression of MuRF1 with MyoD1 or Myogenin resulted in additional cooperative repression of reporter gene activity. To further characterize the role of MuRF1 in the repression of Dusp4 expression, a MuRF1 RING domain mutant was created. The MuRf1 RING mutant was overexpressed in combination with MyoD1 or Myogenin and Dusp4 reporter gene activity was assessed. The MuRF1 RING mutant failed to cooperate with the MRFs to repress the Dusp4 reporter gene, suggesting that ubiquitin ligase activity is required for MuRF1 transcriptional regulatory effects. These data offer evidence that MuRF1 may act as a transcriptional modulator of atrophy‐induced gene expression.
The nuclear protein Fam60a is essential for proper her6 and ascl1b expression and thalamus development in zebrafish (LB825)
Authors: Ricciardi Filomena , Patra Chinmoy , Engel Felix
Published: June 2021

Vertebrate organogenesis is a highly complex and strictly orchestrated process which is dependent on the correct spatiotemporal expression of genes. Gene expression can be regulated by a variety of proteins such as histone acetyltransferases and histone deacetylases; however the conduction of these processes during organ development is still poorly understood. The small nuclear protein FAM60A has been described to be a component of the SIN3‐HDAC1 complex regulating gene expression via histone deacetylation. Here we describe the role of Fam60a during zebrafish development where its depletion leads to severe brain and heart defects. Hdac1 has been shown to promote proneural gene expression such as ascl1b and neurod4. We find that morpholino‐mediated loss of fam60a causes expanded her6 and reduced ascl1b expression in the zebrafish brain. Furthermore, we show that FAM60A possesses a functional Nuclear Localization Signal (NLS) which is essential for its correct function in brain development.

Grant Funding Source: Universitätsklinikum Erlangen
The effect of exercising intensity on learning and memory in humans (LB828)
Authors: Doyle Brendan , Zimmer Mary
Published: June 2021

There is conflicting research regarding the effect of exercise on learning and memory; some studies show no effect, enhanced effects or impaired effects on learning and memory. The objective of this study was to determine if there was an optimal target heart rate range which would provide the greatest improvements in learning and memory. Human subjects (ages 18‐35) were divided into 4 groups: control, low intensity (35‐50% of maximum heart rate (MxHR)), medium intensity (50‐65% MxHR), and high intensity (65‐80% MxHR). Subjects performed a 30min exercise bout on an ergometer within their specified HR range or controls rested. Verbal and visual memory tests were given immediately following the exercise (or rest period in controls), and again 30min later to measure long term memory recall. Heart rate, blood pressure and perceived rate of exertion were measured throughout the protocol. Preliminary results suggest that medium intensity exercise improves immediate learning of verbal associations and the retrieval of word associations from long term storage compared to controls. High intensity exercise appears to impair learning new verbal associations immediately after exercise, but retrieval of long term memory was not affected compared to controls. Finally, preliminary results suggest that medium intensity exercise may slightly improve the ability to recall visual information. These results suggest that high intensity exercise impairs learning and memory, whereas moderate exercise may be beneficial in terms of enhancing memory.

Grant Funding Source: Supported by a Ferris Faculty Research Grant
Effect of upper body resistance training on memory (LB829)
Authors: Jackowski Aaron , Harp Sarah , Zimmer Mary
Published: June 2021

Multiple studies have shown that cardiovascular exercise improves both short and long‐term memory retention, however, very few studies have examined the effect of resistance training or callisthenic exercises on short and long‐term memory. The objective of this study was to determine if upper body resistance training, otherwise known as weight lifting, and calisthenics, which are physical exercises using one’s natural body weight, possess the ability to enhance cognition performance and improve memory retention in humans. Human subjects were placed into either an exercise or control group. The exercise group underwent 3 rounds of upper‐body weight lifting/calisthenics circuit training; each round included a max set of pull‐ups, ten bicep curls per arm, 15 standing dumbbell upright rows, 10 bent over one arm dumbbell rows per arm, 15 dumbbell bench‐press, and a max set of pushups. Exercise subjects had a break of 2 minutes between each round and then repeated the circuit two more times. After exercise, the participants were given a series of verbal and visual memory tests. After a 30 minute rest period another series of verbal and memory tests were given to measure delayed recall. The memory tests measured immediate recall and delayed recall using a verbal paragraph test, and the visual memory test measured immediate and delayed content and spatial memory. Control subjects took the memory tests separated by a 30 minute rest period. Preliminary results suggest that upper‐body resistance training improves both immediate and delayed recall of verbal information. The preliminary results also suggest that upper‐body resistance training may enhance visual content memory but spatial memory was not affected. These results suggest that upper body resistance training may improve learning and memory.

Grant Funding Source: Supported by a Ferris Faculty Research Grant
G protein‐coupled estrogen receptor (GPER) mediates the inhibitory action on P2Y receptor‐mediated Ca2+ signalling and cytokine secretion in human bronchial epithelia (LB831)
Authors: HAO Yuan , CHOW Wai Ming Alison , YIP Chung Yin Wallace , CHAN Wood Yee Woody , CHENG Hon Ki Christopher , KO Wing Hung
Published: June 2021

The airway epithelium plays a central role in respiratory physiology through its transport and immunological functions. Our previous study suggested that P2Y purinergic receptors are expressed in airway epithelia and play a significant role in regulating transepithelial ion transport. P2Y receptors are G protein‐coupled receptors classically signal through Gq, resulting in an increase of intracellular Ca2+ concentration and activation of Ca2+‐dependent downstream signaling pathway. In addition, P2Y receptors are also involved in asthmatic inflammation.

Estrogen (E2) is an important hormone in human physiology. In addition to the classical nuclear hormone receptors ERα and ERβ, a novel estrogen receptor, G‐protein coupled estrogen receptor (GPER/GPR30), was recently identified and found to be involved in both rapid signaling and transcriptional regulations. The action of GPER is unclear but it has been implicated in mediating anti‐inflammatory responses. Our study aimed to investigate the pro‐inflammatory action of P2Y receptors and the interaction between GPER and P2Y receptor‐mediated signaling pathways.

Our results demonstrated the expression and localization of GPER in both primary normal human bronchial epithelial cells (obtained from ScienCell Research Laboratories, San Diego, CA, USA) and 16HBE14o‐ cell line using qPCR, western blotting, and immunofluorescence. Activation of GPER by E2 or its specific agonist, G1, rapidly attenuated a nucleotide‐evoked Ca2+ increase, while the specific GPER antagonist, G15, reversed this GPER‐mediated inhibition. Furthermore, E2 and G1 also inhibited nucleotide‐induced cytokine release. Our results provide the first evidence that human bronchial epithelia express GPER, which interact with the P2Y receptor‐mediated calcium signalling pathway and cytokine secretion. Moreover, the anti‐inflammatory role of GPER may be due to its opposing effect on the pro‐inflammatory pathway activated by the P2Y receptors in inflamed airway epithelia.

Grant Funding Source: Supported by Research Grant Council General Research Fund (#2140730) awarded to W. H. Ko.
Unoprostone protects against cell death, reactive oxygen species, and apoptosis in a model retinitis pigmentosa system using expression of wild type and mutant rhodopsin (LB832)
Authors: Cuppoletti John , Chakrabarti Jayati , Malinowska Danuta , Ueno Ryuji
Published: June 2021

Objective. This study was to determine if unoprostone protects against deleterious effects evident in retinitis pigmentosa (RP). With the P23H mutation in rhodopsin, it is a dominant negative disease, causing the unfolded protein response (UPR) and ER stress. This leads to reactive oxygen species (ROS) production, apoptosis and ultimately cell death.

Methods. Flp‐In T‐Rex‐293 cells were transfected with wild type (WT) or P23H mutant (MUT) rhodopsin in pcDNA5/FRT/TO with pOG44 under control of a tetracycline (tet) promoter. Cell growth, ROS and caspase were measured in non, mock, WT and MUT transfected cells with 1 µg/ml tet ± 100 nM unoprostone. ROS was measured with (5&6)‐chloromethyl‐2’,7’‐dichlorodihydrofluorescein diacetate,acetyl ester and caspase was detected using FITC‐VAD‐FMK.

Results. There were no significant differences between non‐, mock‐ and WT (WT) cells ± unoprostone. At 120 h (n=3) the number of WT cells increased from 54.6 ± 1.2 to 111.7 ± 3.1, while the number of MUT cells decreased from 56.7 ± 3.6 to 19.0 ± 1.3. Unoprostone increased significantly (P<0.005) the number of MUT cells to 32.0 ± 0.5. ROS (FU, n=6) increased from 332.9 ± 0.7 in WT cells to 2816.8 ± 6.4 in MUT cells and this was significantly decreased (P<0.0005) to 1331.0 ± 3.0 with unoprostone. Caspase (FU, n=6) at 120 h increased from 365.7 ± 5.3 in WT cells to 895.5 ± 10.2 in MUT cells and this was significantly reduced (P<0.0005) to 523.2 ± 11.3 with unoprostone.

Conclusions. The data shows that unoprostone can slow the pathological events (ER stress, ROS production, and apoptosis) and cell death seen in RP with UPR. Increased ROS generation and apoptosis are also seen in age related macular degeneration (AMD), suggesting unoprostone should also be investigated in dry AMD.

Grant Funding Source: Supported by Sucampo AG
Metastatic tumor cells induce a crosstalk between endothelial p38 and ERK1/2 MAPK pathways activating endothelial MMP system (LB838)
Authors: Glinskii Olga , Glinsky Vladislav
Published: June 2021

Our recent results demonstrate that metastatic cell adhesive interactions with endothelial cells (EC) are stabilized by the endothelial integrin α3β1 leading to the activation of p38 and ERK1/2 pathways in endothelial cells when they interact with tumor cells. Here we demonstrate that, while FAK/Src/RhoA/ROCK/MKK3/6/p38 pathway is fully activated downstream of α3β1 in EC, endothelial ERK1/2 is progressively dephosphorylated despite time‐dependent MEK1/2 activation. We further demonstrate that ERK1/2 dephosphorylation results from a crosstalk between p38 and ERK1/2 pathways whereby p38‐dependent increase in the expression of dual specificity phosphatase MKP‐3 causes progressive ERK1/2 deactivation. The resulting outcome of this complex crosstalk between the two major endothelial MAP kinases is p38‐dependent activation of MT1‐MMP, increase in MMP‐9 expression, and progressive release of MMP‐2 from ECs. We conclude that upon interaction with ECs, metastatic cells highjack EC MMP system via α3β1‐induced crosstalk between major MAPK pathways to facilitate tumor cell extravasation.

Grant Funding Source: Supported by the VA BLR&D Service Award 1I01BX000609 and NIH Grant R01CA160461 to VV Glinsky
High glucose stress induces mesenchymal stromal cell senescence through up‐regulating autophagy (LB842)
Authors: Chang Tzu‐Ching , Hsu Min‐Fen , Wu Kenneth K.
Published: June 2021

Diabetic hyperglycemia was reported to induce impairment of human bone marrow (BM) hematopoietic function and cause niche dysfunction. As multi‐potent mesenchymal stromal cells (MSC) are a key constituent of BM hematopoietic niche, we determined the influence of high glucose (HG) on cultured MSCs. MSCs were cultured in medium containing 5.5 mM glucose (low glucose, LG) or 25 mM glucose (HG) for up to 28 days and cell proliferation and morphology were analyzed. MSCs cultured under HG exhibited reduced cumulative population doubling and BrdU incorporation and increased p16 and p21 in a time‐dependent manner. Senescence‐associated β‐galactosidase (SA‐β‐Gal) positive cells and interleukin‐6 (IL‐6) production were increased. Cells underwent morphological changes consistent with senescence. Since cellular senescence is controlled by autophagy, we determined whether HG suppresses autophagy thereby inducing premature senescence. Interestingly, MSC cultured in HG medium exhibited enhancement of beclin‐1 expression and LC3‐II generation when compared to MSC cultured in LG. HG increased beclin‐1 (Atg 6) as well as Atg 5, Atg 7, and Atg 12 mRNA when compared to LG. HG also significantly increased autophagosome formation. To elucidate the relationship between autophagy and senescence, we treated MSCs with 3‐methyladenine (3‐MA), an inhibitor of autophagy and analyzed changes in senescence. 3‐MA prevented growth arrest and p21 elevation and abrogated SA‐β‐Gal and IL‐6 expressions. These results indicate that HG induces senescence through activation of autophagy. It was reported that HG induces cell injury via generation of reactive oxygen species (ROS). We evaluated the effect of N‐acetylcysteine (NAC), an antioxidant, on MSC autophagy and senescence. NAC reduced Atg expressions, LC3‐II generation and autophagosome formation accompanied by preventing senescence. DPI, an inhibitor of NADPH oxidase, exerted a comparable effect as NAC.

These findings indicate that HG induces premature senescence by ROS‐mediated autophagy upregulation. MSC senescence contributes to niche dysfunction and bone marrow inflammation.
Virtual Humans and Avatars ‐ Healthcare Personal Agents in the Palm of Your Hand (LB855)
Authors: Federico Niksha , Federico Tessya , Federico Francesca , Sternik Gabriel , Esmaeli‐Azad Babak
Published: June 2021

The advent of wireless sensors and miniaturized wearables have made the possibility of home diagnostics prior to a doctor visit and connected healthcare a reality. Engagement of everyday people for self diagnosis, however, remains a continued challenge as in most cases the much needed user friendly and engaging user interface for wireless devices are lacking.

Recent advancements in the fields of physiology, psychology, artificial intelligence, speech recognition, natural language learning, computer graphics, gamification and cinematography have made the development of virtual human/ avatar personal health agents that are engaging, wireless smart phone and tablet native, and personalized possible.

We here describe development of a personalized virtual human/ avatar health agent for incentivized, gamified, home self testing of 21 indications using FDA approved miniaturized wearable medical diagnostic devices. The avatars are designed to have multiple characters capable of understanding 15 different languages. Data obtained from patient interactions and device testing is stored in the cloud for analysis. Aesthetics of patient engagement and gamification will be presented. Development of disease specific scripts to determine different stages of a particular disorder including live physician telemedicine capabilities and connected healthcare will be described. Our findings are used as a solution for the development of team SMART McCoy's solution for the Qualcomm Medical Tricorder X Prize competition. In full development this system will assist in lessening the unnecessary primary care doctor visits (i.e., more than $10 billion annual cost in US alone) and provides the mean for making connected healtcare and precision medicine a reality.
Metabolics of PH – an update
Authors: Hughes Allyson L. , Hemnes Anna R.
Published: June 2021

Purpose of review

While there has been a longstanding interest in metabolic disease in pulmonary hypertension, publications in the last several years have translated basic science findings to human disease and even led to recently published studies of metabolic therapy in pulmonary arterial hypertension that are discussed here.

Recent findings

Progress has been made in four key areas including mechanisms of insulin resistance in pulmonary arterial hypertension, the role of obesity in pulmonary vascular disease, novel clinical trials targeting metabolism in pulmonary hypertension, and the role of metabolism in chronic thromboembolic pulmonary hypertension.

: Insulin resistance in pulmonary arterial hypertension is primarily in the lipid axis. There are systemic manifestations of insulin resistance including right ventricular lipotoxicity. Obesity is associated with elevation of right ventricular systolic pressure even in a healthy population and therapies in pulmonary arterial hypertension that target metabolism hold promise for improving exercise, right ventricular function, and visceral adiposity. Finally, there are emerging data that chronic thromboembolic pulmonary hypertension is similarly characterized by metabolic alterations, though the specific metabolites may be different from pulmonary arterial hypertension.
Effects of Gluten-free Diet on Lipid Profile Risk Factors in Celiac Children
Authors: Angi Alessia , Polidori Nella , Cerruto Marina , Chiarelli Francesco , Mohn Anna Angelika
Published: June 2021

Many studies raise concerns about the nutritional consequences of gluten-free diet. It has been documented that gluten-free (GF) foods have a higher glycemic index, saturated fats, and lower content of micronutrients determining important health implications. In this retrospective study, we evaluated the change in cardiometabolic risk factors in prepubertal celiac children in remission following different gluten-free diet regimes. Patients using processed GF foods showed a significant increase over time for standard deviation score-total cholesterol, standard deviation score-low density lipoprotein cholesterol, and fasting glycemia. These alterations were not confirmed in patients using naturally GF foods. Therefore, processed GF foods might promote unfavorable alterations of metabolic parameters, especially those associated with increased risk of cardiovascular diseases. Supervision of a dietitian and medical practitioner is recommended to ensure nutritional adequacy and monitoring of cardiovascular risk factors in this population.
Urine Test Predicts Kidney Injury and Death in COVID-19
Authors: Xu Katherine , Shang Ning , Levitman Abraham , Corker Alexa , Kudose Satoru , Yaeh Andrew , Neupane Uddhav , Stevens Jacob , Mohan Sumit , Sampogna Rosemary , D’Agati Vivette , Kiryluk Krzysztof , Barasch Jonathan
Published: June 2021

Background

Kidney injury is common in COVID-19 infection, but serum creatinine (SCr) is not a sensitive or specific marker of kidney injury. We hypothesized that molecular markers of tubular injury could diagnose COVID-19 associated kidney damage and predict its clinical course.

Methods

This is a prospective cohort study of 444 consecutive COVID-19 patients (43.9% females, 20.5% African American, 54.1% Latinx) in Columbia University’s Emergency Department at the peak of the New York pandemic (March-April 2020). Urine and blood were collected simultaneously at admission (median time of day 0, IQR 0-2 days) and within 1 day of a positive SARS-CoV-2 test in 70% of patients. Biomarker assays were blinded to clinical data.

Results

Urinary NGAL (uNGAL) was strongly associated with AKI diagnosis (267±301 vs. 96±139 ng/mL, P=1.6×10−10). uNGAL >150ng/mL had 80% specificity and 75% sensitivity to diagnose AKIN stage 2 or higher. uNGAL quantitatively predicted the duration of AKI and outcomes, including death, dialysis, shock, and longer hospital stay. The risk of death increased 73% per standard deviation of uNGAL [OR (95%CI): 1.73 (1.29-2.33), P=2.8×10−4] and was independent of baseline SCr, co-morbidities, and proteinuria [adjusted OR (95%CI): 1.51 (1.10-2.11), P=1.2×10−2]. Proteinuria and uKIM-1 also indicated tubular injury but were not diagnostic of AKI. Typically, distal nephron segments transcribe NGAL, but in COVID-19 biopsies with widespread acute tubular injury (ATI), NGAL expression overlapped KIM-1 in proximal tubules.

Conclusion

uNGAL predicted diagnosis, duration, and severity of AKI and ATI, as well as hospital stay, dialysis, shock, and death in patients with acute COVID-19.
SARS-CoV-2 shedding, infectivity and evolution in an immunocompromised adult patient
Authors: Mendes-Correa Maria Cassia , Ghilardi Fábio , Salomão Matias Chiarastelli , Villas-Boas Lucy S. , Vicente de Paula Anderson , Tozetto-Mendoza Tania Regina , Freire Wilton , Sales Flavia C. , Romano Camila M. , Claro Ingra M. , de Souza Leandro Menezes , Ramos Jessica F. , Paiao Heuder Gustavo Oliveira , Szor Roberta Shcoinik , Kallas Esper Georges , Sabino Ester C. , Witkin Steven S. , Faria Nuno R.
Published: June 2021

This report describes a persistent SARS-CoV-2 infection of at least 218 days in a male, in his 40’s who had undergone a prior autologous hematopoietic stem cell transplant due to a diffuse large B-cell lymphoma. He did not manifest a humoral immune response to the virus. Whole-genome sequencing and viral cultures confirmed a continual infection with a replication-positive virus that had undergone genetic variation for at least 196 days following symptom onset.
Natural language inference for clinical registry curation
Authors: Percha Bethany , Pisapati Kereeti , Gao Cynthia , Schmidt Hank
Published: June 2021

Clinical registries - structured databases of demographic, diagnosis, and treatment information for patients with specific diseases or phenotypes - play vital roles in high-quality retrospective studies, operational planning, and assessment of patient eligibility for research, including clinical trials. However, registries are extremely time and resource intensive to curate. Natural language processing (NLP) can help, but standard NLP methods require specially annotated training sets or the construction of separate models for each of dozens or hundreds of different registry fields, rendering them insufficient for registry curation at scale. Natural language inference (NLI), a specific branch of NLP focused on logical relationships between statements, presents a possible solution, but NLI methods are largely unexplored in the clinical domain outside the realm of conference shared tasks and computer science benchmarks. Here we convert registry curation into an NLI problem, applying five state-of-the-art, pretrained, deep learning based NLI models to clinical, laboratory, and pathology notes to infer information about 43 different breast oncology registry fields. We evaluate the models’ inferences against a manually curated, 7439 patient breast oncology research database. The NLI models show considerable variation in performance, both within and across registry fields. One model, ALBERT, outperforms the others (BART, RoBERTa, XLNet, and ELECTRA) on 22 out of 43 fields. A detailed error analysis reveals that incorrect inferences primarily arise through models’ misinterpretations of temporality--they interpret historical findings as current and vice versa--as well as confusion based on subtle terminology and abbreviation variants common in clinical notes. However, modern NLI methods show promise for increasing the efficiency of registry curation, even when used “out of the box” with no additional training.
A pilot feasibility study on SARS-CoV-2 detection method based on nasopharyngeal lavage fluid
Authors: Frezza Daniele , Fabbris Cristoforo , Franz Leonardo , Vian Elisa , Rigoli Roberto , De Siati Rosalba , Emanuelli Enzo , Luigi Bertinato , Boscolo-Rizzo Paolo , Spinato Giacomo
Published: June 2021

Objective

Nose and nasopharyngeal swab is the preferred and worldwide accepted method to detect the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) within the nose and nasopharynx. This method may be linked with possible difficulties, such as patient’s discomfort or complications. This paper shows a pilot study of SARS-CoV-2 detection with nasal and nasopharyngeal lavage fluids.

Methods

Nasal lavage fluid was collected from patients who were submitted to SARS-CoV-2 screening test, due to a preceding positive rapid antigen test. A control group was enrolled among healthcare professionals whose nasopharyngeal swab tested negative. Nasal lavages were performed using isotonic saline solution injected through a nasal fossa. Both lavage fluid and traditional nasopharyngeal swab were analyzed by real-time PCR and antigenic test.

Results

A total of 49 positive subjects were enrolled in the study. Results of the analysis on lavages and nasopharyngeal swabs were concordant for 48 cases, regardless of the antigenic and molecular test performed. RT-PCR resulted weakly positive at swab in one case and negative at lavage fluid. Among the control group (44 subjects) nasopharyngeal swab and lavage fluid analyses returned a negative result. Sensitivity of the molecular test based on nasal lavage fluid, compared to traditional nasal swab, was 97.7%, specificity was 100%, and accuracy was 98.9%, with high agreement (Cohen’s k, 0.978).

Conclusion

Nasal and nasopharyngeal lavages resulted to be highly reliable and well tolerated. A larger series is needed in order to confirm these results. This approach may potentially represent a valid alternative to the traditional swab method in selected cases.
A palmitoylation code controls PI4KIIIα complex formation and PI(4,5)P2 homeostasis at the plasma membrane
Authors: Batrouni Alex G. , Bag Nirmalya , Phan Henry , Baird Barbara A. , Baskin Jeremy M.
Published: June 2021

PI4KIIIα is the major enzyme responsible for generating the phosphoinositide PI(4)P at the plasma membrane. This lipid kinase forms two multicomponent complexes, both including a palmitoylated anchor, EFR3. Whereas both PI4KIIIα complexes support production of PI(4)P, the distinct functions of each complex and mechanisms underlying the interplay between them remain unknown. Here, we present roles for differential palmitoylation patterns within a tri-Cys motif in EFR3B (Cys5/Cys7/Cys8) in controlling the distribution of PI4KIIIα between these two complexes at the plasma membrane and corresponding functions in phosphoinositide homeostasis. Spacing of palmitoyl groups within three doubly palmitoylated EFR3B “lipoforms” affects both its interactions with TMEM150A, a transmembrane protein governing formation of a PI4KIIIα complex functioning in rapid PI(4,5)P2 resynthesis following PLC signaling, and its partitioning within liquid-ordered and -disordered regions of the plasma membrane. This work identifies a palmitoylation code in controlling protein–protein and protein–lipid interactions affecting a plasma membrane-resident lipid biosynthetic pathway.

SUMMARY STATEMENT

Different palmitoylation patterns on a lipid kinase adaptor protein control partitioning of the kinase between two spatiotemporally and functionally distinct complexes within the plasma membrane.
Expanding Clinical Phage Microbiology: Simulating Phage Inhalation for Respiratory Tract Infections
Authors: Porat Shira Ben , Gelman Daniel , Yerushalmy Ortal , Alkalay-Oren Sivan , Coppenhagen-Glazer Shunit , Cohen-Cymberknoh Malena , Kerem Eitan , Amirav Israel , Nir-Paz Ran , Hazan Ronen
Published: June 2021

Phage therapy is a promising antibacterial strategy for resistant respiratory tract infections. Phage inhalation may serve this goal; however, it requires a careful assessment of their delivery by this approach. Here we present anin-vitromodel to evaluate phage inhalation.

Eight phages, most of which target CF-common pathogens, were aerosolized and administered to a real-scale CT□derived 3D airways model with a breathing simulator. Viable phage loads reaching the output of the nebulizer and the tracheal level of the model were determined and compared to the loaded amount.

Phage inhalation resulted in a diverse range of titer reduction, primarily associated with the nebulization process. No correlation was found between phage delivery to the phage physical or genomic dimensions. These findings highlight the need for tailored simulations of phage delivery, ideally by a patient-specific model in addition to proper phage matching, to increase the potential of phage therapy success.

Take-Home Message

Phage therapy can be used against infectious diseases if personally tailored. Using a 3D airways model, we show that phage delivery by inhalation to the respiratory tract is unpredictable and also requires a precise evaluation.
OsciDrop: A Versatile On-demand Droplet Generator
Authors: Ye Shun , Zheng Xu , Li Caiming , Huang Weihang , Tao Yi , Yu Yanghuan , Yang Limin , Lan Ying , Ma Liang , Bian Shengtai , Du Wenbin
Published: June 2021

Droplet microfluidics is a powerful tool in many biological and clinical applications. Microfluidic chips, such as flow-focusing droplet generators, have been extensively used to high-throughput encapsulate reactions with single-cell and single-molecular resolutions. However, microfabrication is expensive and precision-demanding, preventing it from widespread use in biomedical laboratories and clinical facilities. Herein, we present a versatile chip-free droplet generator, OsciDrop, for generating size-tunable droplets on demand, with high uniformity. OsciDrop segments the fluid flowing out of the orifice of a micropipette tip into droplets by oscillating the tip under the surface of a continuous oil phase. We investigated the factors influencing droplet generation by examining several control parameters. Results show that flow rate, oscillating amplitude, and frequency are key parameters to generate monodisperse droplets on demand. And OsciDrop is able to generate droplets in a flexible and repeatable manner. Importantly, using an optimal asymmetrical oscillation waveform, OsciDrop can controllably generate monodisperse droplets spanning a wide volume range (200 pL - 2 μL). To demonstrate the ability of OsciDrop for chip-free droplet assays, a digital loop-mediated isothermal amplification (dLAMP) was performed to absolutely quantify African swine fever virus (ASFV). The OsciDrop method opens up a feasible and versatile avenue to perform droplet-based assays, exhibiting full accessibility for chip-free droplet microfluidics.
Differential repression of Otx2 underlies the capacity of NANOG and ESRRB to induce germline entry
Authors: Vojtek Matúš , Zhang Jingchao , Sun Juanjuan , Zhang Man , Chambers Ian
Published: June 2021

Primordial germ cells (PGCs) are induced in the embryo by signals, including BMP emanating from extra-embryonic ectoderm, that act on cells in the post-implantation epiblast. PGC development can be recapitulated in vitro through the exposure of epiblast-like cells (EpiLCs) to appropriate cytokines, resulting in differentiation into PGC-like cells (PGCLCs). Interestingly, the requirement for cytokines to induce PGCLCs can be bypassed by enforced expression of the transcription factor (TF) NANOG. However, the underlying mechanisms are not fully elucidated. Here, we show that Otx2 downregulation is essential to enable NANOG to induce PGCLC formation. Moreover, while previous work has shown that the direct NANOG target gene Esrrb can substitute for several functions of NANOG, enforced expression of ESRRB cannot promote PGCLC specification from EpiLCs. This appears to be due to differential downregulation of Otx2 by NANOG and ESRRB, since induction of ESRRB in Otx2+/- EpiLCs activates expression of the core PGC TFs, Blimp1, Prdm14 and Ap2γ and emergence of PGCLCs. This study illuminates the interplay of TFs occurring at the earliest stages of PGC specification from a state of competence.
X-ray induced bio-acoustic emissions from cultured cells
Authors: Matarèse Bruno F.E. , Rahmoune Hassan , Vo Nguyen T. K. , Seymour Colin B. , Schofield Paul N. , Mothersill Carmel
Published: June 2021

Purpose

We characterise for the first time the emission of bio-acoustic waves from cultured cells irradiated at doses of X-ray photon radiation relevant to medical and accidental exposure.

Methods and materials

Human cancer cell lines (MCF-7, HL-60) and control cell-free media were exposed to 1 Gy X-ray photons while recording the sound generated before, during and after irradiation. Cellular cytotoxicity following photon irradiation was determined by extracellular LDH levels, and irradiated cell conditioned media were tested for their ability to elicit a bystander effect in reporter cells.

Results

We report the first recorded acoustic signals captured from a collective pressure wave response to ionising irradiation. The signature of the collective acoustic peaks was temporally wider and with higher acoustic power for irradiated HL-60 than for irradiated MCF-7.

Conclusions

We show that at doses of X-ray irradiation capable of producing the release of bystander effect-inducing activity, both cell types emit a characteristic acoustic signal for the duration of the radiation pulse. The rapid signal decay is consistent with a passive rather than an active acoustic signal generation. This preliminary study suggests that further work on the potential role of radiation induced acoustic emission (RIAE) in the cellular bystander effect is merited.
Managing African Swine Fever: Assessing the Potential of Camera Traps in Monitoring Wild Boar Occupancy Trends in Infected and Noninfected Zones, Using Spatio-temporal Statistical Models
Authors: Bollen Martijn , Neyens Thomas , Fajgenblat Maxime , De Waele Valérie , Licoppe Alain , Manet Benoît , Casaer Jim , Beenaerts Natalie
Published: June 2021

The recent spreading of African swine fever (ASF) over the Eurasian continent has been acknowledged as a serious economic threat for the pork industry. Consequently, an extensive body of research focuses on the epidemiology and control of ASF. Nevertheless, little information is available on the combined effect of ASF and ASF-related control measures on wild boar (Sus scrofa) population abundances. This is crucial information given the role of the remaining wild boar that act as an important reservoir of the disease. Given the high potential of camera traps as a non-invasive method for ungulate trend estimation, we assess the effectiveness of ASF control measures using a camera trap network. In this study, we focus on a major ASF outbreak in 2018-2020 in the South of Belgium. This outbreak elicited a strong management response, both in terms of fencing off a large infected zone as well as an intensive culling regime. We apply a Bayesian multi-season site-occupancy model to wild boar detection-nondetection data. Our results show that (1) occupancy rates at the onset of our monitoring period reflect the ASF infection status; (2) ASF-induced mortality and culling efforts jointly lead to decreased occupancy over time; and (3) the estimated mean total extinction rate ranges between 22.44% and 91.35%, depending on the ASF infection status. Together, these results confirm the effectiveness of ASF-control measures implemented in Wallonia (Belgium), which has regained its disease-free status in December 2020, as well as the usefulness of a camera trap network to monitor these effects.
Longitudinal Developmental Trajectories Do Not Follow Cross-Sectional Age Associations in Hippocampal Subfield and Memory Development
Authors: Keresztes Attila , Raffington Laurel , Bender Andrew R. , Bögl Katharina , Heim Christine , Shing Yee Lee
Published: June 2021

Many cross-sectional findings suggest that volumes of specific hippocampal subfields increase in middle childhood and early adolescence. In contrast, a small number of available longitudinal studies observed decreased volumes in most subfields over this age range. Further, it remains unknown whether structural changes in development are associated with corresponding gains in children’s memory. Here we report cross-sectional age differences in children’s hippocampal subfield volumes together with longitudinal developmental trajectories and their relationships with memory performance. In two waves, 109 healthy participants aged 6 to 10 years (wave 1: MAge=7.25, wave 2: MAge=9.27) underwent high-resolution magnetic resonance imaging to assess hippocampal subfield volumes, and completed cognitive tasks assessing hippocampus dependent memory processes. We found that cross-sectional age-associations and longitudinal developmental trends in hippocampal subfield volumes were highly discrepant, both by subfields and in direction. Further, volumetric changes were largely unrelated to changes in memory, with the exception that increase in subiculum volume was associated with gains in spatial memory. Importantly, the observed longitudinal patterns of brain-cognition coupling could not be inferred from cross-sectional findings. We discuss potential sources of these discrepancies. This study underscores that children’s structural brain development and its relationship to cognition cannot be inferred from cross-sectional age comparisons.

Highlights

The subiculum undergoes volumetric increase between 6-10 years of age

Change across two years in CA1-2 and DG-CA3 was not observed in this age window

Change across two years did not reflect age differences spanning two years

Cross-sectional and longitudinal slopes in stark contrast for hippocampal subfields

Longitudinal brain-cognition coupling cannot be inferred from cross-sectional data