bluuebunny/crossref_metadata_2025 huggingface dataset:
Testing a Feedback‐controlled Heated Vest to Treat Thermoregulatory Dysfunction in Persons with Spinal Cord Injury
Authors: Ortiz Vasquez Luis , Parada Wilber , Polizzi Jessica , Lee Ingrid , Bang Charlene , Bauman William , Handrakis John P.
Published: May 2022

Study Objective

After spinal cord injury (SCI), The ability to maintain a constant core body temperature (Tcore) is compromised. Persons with higher levels of SCI (Hi‐SCI: above the first thoracic vertebrae) are far more vulnerable to hypothermia (Tcore <35°C) when exposed to cool ambient temperatures. A decrease in Tcore in persons with Hi‐SCI has been associated with impaired cognitive performance after limited exposure to cool environmental conditions (2 hours at 18°C). The objective of this pilot study is to determine the efficacy of a feedback‐controlled heated vest (HV) to minimize the decrease in Tcore and associated cognitive impairment in persons with Hi‐SCI when exposed to cool ambient conditions.

Methods

Three participants with Hi‐SCI (C4‐C7, AIS A‐B) were recruited to study under two conditions [HV and non‐heated vest (NHV)] in random order on 2 separate visits when exposed to a cool challenge (CC). To obtain Tcore in a continuous manner, participants ingested a CorTempR capsule 1‐3 hours before initiation of the study. Abdominal (Tabd) and distal (Tdsk) skin temperatures were measured. Fifteen minutes of baseline (BL) data were collected while seated at thermoneutral (25°C). At the end of BL, a cognitive battery that consisted of the Montreal Cognitive Assessment (MoCA) and Stroop Color and Word Test (SCWT) was administered. Participants were then fitted with either HV or NHV and transferred into an 18°C room for 2 hours, after which the cognitive battery was once again administered.

Results: During Cool Challenge (values are expressed as BL

‐ CC): The decrease in Tcore was less when wearing the HV (‐0.22°C) than when wearing the NHV (‐0.54°C). The increase in Tabd was greater when wearing the HV (4.34°C) than when wearing the NHV (0.90°C). When wearing the HV, Tdsk of the fingers was higher (6.48°C HV vs. 4.20°C NHV), but not of the toes (6.46°C HV vs. 7.92°C NHV). The decrease in performance for the MoCA was less when wearing the HV (‐8.33%) than when wearing the NHV (‐43.33%). The decrease in score for the Stroop word test was less when wearing the HV (‐0.17%) than when wearing the NHV (‐3.32%). Of note, the Stroop Color and Color‐Word score increased when wearing the HV (10.12% and 5.73%, respectively) but decreased when wearing the NHV (‐0.97% and ‐4.9%, respectively).

Conclusion

In persons with Hi‐SCI, our preliminary findings demonstrate the ability of the HV to minimize the expected decline in Tcore when exposed to the CC and decreased the decline in cognitive performance. While our results are quite encouraging, they require confirmation in a larger number of participants. However, once confirmed, our HV may offer persons with Hi‐SCI protection against adverse declines in Tcore when exposed to cool environmental conditions.
Pre‐fatiguing Isometric Quadriceps Exercise Impairs Contralateral Quadriceps W’ During All‐out and Not Target Torque Time to Task Failure Exercise
Authors: Lewis Matthew T. , Broxterman Ryan M. , Craig Jesse C. , Weavil Joshua C. , Laginestra Fabio G. , Jarrett Catherine L. , Bunsawat Kanokwan , Amann Markus , Richardson Russell S.
Published: May 2022

Purpose

A crossover effect of pre‐fatiguing dynamic quadriceps contractions, between contralateral legs, was recently documented, evidencing a centrally mediated negative impact on performance, assessed by time to task failure (TTF) at a target intensity. This has since been reproduced, however, whether a crossover effect exists during isometric exercise, which permits exercise in a restricted space for a more intricate assessment of muscle function, for example, by magnetic resonance imaging (MRI), has not been determined. Therefore, this study interrogated the crossover effect of isometric pre‐fatiguing quadriceps exercise with the hypothesis that a crossover would impair TTF during isometric exercise.

Methods

Eight healthy participants (5M/3F) performed all‐out isometric exercise tests (60 maximal voluntary contractions, 3 s contraction, 2 s relaxation) to determine the impulse above end test torque (W′) and critical torque (CT) to establish a target torque that invoked a TTF between 8‐15 min, as defined by the torque‐duration relationship (TTF = W’/(target torque – CT)). Once established, target torque TTF and all‐out exercise CT and W’ were measured with and without pre‐fatiguing all‐out quadriceps exercise in the contralateral leg to assess the crossover effect.

Results

TTF averaged 9.8 ± 0.4 min and, interestingly, was not altered by pre‐fatigue in the contralateral leg (11.9 ± 1.0 min, p = 0.34). CT was not different in control vs. pre‐fatigue (73 ± 8 vs. 76 ± 10 N*m, p= 0.34) but W′, an indicator of exercise capacity, was reduced by 25% (6200 ± 1200 vs. 4700 ± 900 N*m*s, p = 0.02).

Conclusions

These data document a crossover effect of pre‐fatiguing isometric quadriceps exercise on subsequent contralateral quadriceps isometric exercise, but only during all‐out exercise and not target torque TTF. Furthermore, as a consequence of the unique experimental approach, utilizing a dual‐leg isometric ergometer designed to fit in a MRI, the negative impact of contralateral limb pre‐fatigue (e.g. increased afferent feedback) on W′ has been documented for the first time.
Examining Vascular Function and Oxidative Stress in Young Individuals with Generalized Anxiety Disorder
Authors: Richardson Jake , Weggen Jennifer , Darling Ashley , Chiu Alex , Decker Kevin , Garten Ryan
Published: May 2022

Generalized anxiety disorder (GAD) is a disabling psychiatric condition that can result in physiological manifestations linked to a greater risk for cardiovascular disease (CVD). Peripheral vascular dysfunction may be one mechanism augmenting CVD risk as GAD is linked to an overactivation of the sympathetic nervous system, a phenomenon that can result in excessive vasoconstriction and increases in oxidant production in the vasculature and bloodstream. Excessive oxidant production can overpower the body’s endogenous antioxidant defense system resulting in oxidative stress, a known modulator of peripheral vascular function.

Purpose

The purpose of this study was to evaluate peripheral vascular function and oxidant production in young individuals with GAD. We hypothesized that young individuals with GAD would exhibit blunted vascular function and greater oxidant production when compared to age‐ and sex‐matched individuals without GAD.

Methods

Fifteen individuals with GAD (11 females, 4 males; age 22 ± 2 years), and 13 healthy controls (CTRL; 9 females, 4 males, 23 ± 4 years), participated in the study. The 7‐item Generalized Anxiety Disorder (GAD‐7) self‐report scale was utilized to screen for GAD with a GAD‐7 score of ≥ 10 used as the cut‐off point for the GAD group and a score of ≤ 5 used as the cut‐off point for the control group. Arm vascular function was assessed via the reactive hyperemia‐induced flow mediated dilation of the brachial artery (BAFMD) technique and evaluated with Doppler ultrasonography. Brachial artery and arm microvascular function were determined by percent change of diameter from baseline normalized for BA shear rate (BAD/Shear) and blood flow area under the curve (BF AUC), respectively. Whole blood superoxide concentration was assessed via electron paramagnetic resonance (EPR) to determine potential differences in oxidant production.

Results

Young individuals with GAD had greater GAD‐7 scores (p < 0.001) when compared to controls (GAD: 15 ± 3; range 12 – 21; CTRL: 2 ± 2; range 0 – 5). BAD/Shear (GAD: 0.11 ± 0.06; CTRL: 0.10 ± 0.05 %/sec; p = 0.6), BF AUC (GAD: 21029 ± 11320; CTRL: 21727 ± 14493 milliliters; p = 0.9), and oxidant production (GAD: 52463 ± 72474; CTRL: 83472 ± 185805; p = 0.6) were not significantly different between groups.

Conclusion

Contrary to our hypothesis, young individuals with GAD demonstrated similar macro‐ and microvascular function as well as similar whole blood oxidant production when compared to age‐ and sex‐matched individuals without GAD. This may highlight preserved cardiovascular health in young individuals with GAD.
Prolactin Directs Branchial and Intestinal Gene Expression of Solute Transporters and Aquaporins in Mummichog (Fundulus heteroclitus)
Authors: Bradley Serena , Puterbaugh Katie , Hageman Annie , Verspyck Adrian , Shaw Lydia , Danielson Libby , Hou Yubo , Breves Jason
Published: May 2022

Mummichogs (Fundulus heteroclitus) can inhabit a wide range of environmental salinities because of their capacity to modulate the expression of genes that encode solute transporters and water channels within key osmoregulatory organs. Over 60 years ago, the pituitary hormone, prolactin (Prl), was identified as an essential factor that enables mummichogs to tolerate freshwater (FW) environments; nonetheless, the molecular targets of Prl signaling in this model teleost remain unknown. In the current study, we determined that two Prl receptors, designated prlra and prlrb, are expressed in osmoregulatory organs in salinity‐dependent fashions. We administered ovine Prl (oPrl; 1 or 5 mg g‐1 body weight) by intraperitoneal injection to mummichogs held in brackish water (12 ppt) and first characterized the expression of genes associated with FW‐type (ion absorptive) and seawater (SW)‐type (ion secretory) ionocytes. Within FW‐type ionocytes, oPrl stimulated the expression of Na+/Cl‐ cotransporter 2, aquaporin 3, and prlra. Branchial Na+/H+ exchanger 2and ‐3 gene expression did not respond to oPrl. The expression levels of genes associated with SW‐type ionocytes, such as Na+/K+/2Cl‐cotransporter 1and cystic fibrosis transmembrane regulator 1, were reduced following oPrl administration. Given the role of aquaporins in supporting fluid absorption by the intestine, we then assessed whether multiple aquaporin encoding genes were responsive to oPrl. We observed that aquaporin 1a and ‐8 were diminished by oPrl. As in the gill, oPrl stimulated the expression of prlra. Our collective data indicate that Prl promotes the survival of mummichogs in FW by simultaneously modulating a suite of genes within the gill and intestine that underlie critical ion‐ and water‐transporting processes.
Evaluating the Association of Sleep & Physical activity with Prolonged Concussion Symptoms
Authors: Neely Linda , Smulligan Katherine L. , Wingerson Mathew J. , Seehusen Corrine , Simon Stacey , Wilson Julie C. , Howell David R.
Published: May 2022

Introduction

Adolescents with concussion will typically experience symptom resolution within 4‐weeks of injury; however, persistent post‐concussion symptoms (PPCS) may develop in approximately 30% of adolescents. Concussion management guidelines currently recommend a brief (24‐48 hour) rest period followed by sub‐symptom exercise to facilitate recovery. Additionally, poor sleep is associated with longer recovery, and early physical activity (PA) may accelerate symptom resolution. By understanding the individual association of both sleep and PA with symptom recovery, a basis can be provided for future recommendations on concussion intervention. Therefore, the purpose of this study was to determine whether sleep characteristics (duration, timing) and/or PA (steps/day, exercise frequency, duration, exercise intensity) in the first month after concussion are associated with PPCS.

Methods

We performed a prospective study of adolescent athletes who sustained a concussion and were evaluated within 14 days of injury. Participants received a Fitbit Charge 3TM to track PA and sleep parameters during recovery. The Post‐Concussion Symptom Inventory (PCSI) was used to determine symptom severity at the initial visit and the clearance visit. Participants were followed until symptom resolution, and the main outcome interest was development of PPCS (symptom duration ≥ 28 days). We then used a logistic regression model to examine associations between PA and sleep parameters with PPCS.

Results

Of the 49 patients included in the analysis, 47% (n=23, symptom resolution=57±23 days post‐injury) developed PPCS and 53% (n=26, symptom resolution=15±7 days post‐injury) did not. There was a significant difference between the PPCS and no PPCS groups in three different variables: steps/day (7526±2975 vs. 9803±3786; p=0.02), exercise days/week (2.5±2.2 vs. 4.4±2.1; p=0.005), and time in bed awake (1.2±0.3 hours/night vs. 0.8±0.3 hours/night; p=0.03). In our multivariable model, the risk of developing PPCS decreased by 1.6x with each additional exercise session/week (adjusted odds ratio=0.616, 95% confidence interval=0.389, 0.974, p=0.038).

Conclusions

The results suggest that participating in multiple exercise sessions per week during concussion recovery is associated with a lower risk of developing PPCS. Furthermore, taking more steps/day and spending less time awake in bed are associated with a shorter recovery period. Counseling patients on optimizing sleep and PA during concussion recovery may be one method for clinicians to promote recovery for their patients.
Dsk2 Induces Aggregation of Polyubiquitinated Proteins
Authors: Mallon Erica L. K. , Kraut Daniel A.
Published: May 2022

The ubiquitin‐proteasome system (UPS) is an intricate system responsible for protein degradation in eukaryotic cells. Within the UPS, proteins targeted for degradation are tagged with polyubiquitin chains and subsequently recognized and degraded by the 26S proteasome. The three proteasome‐associated shuttle proteins Dsk2, Rad23, and Ddi1 mediate substrate degradation by helping deliver ubiquitin‐tagged substrates to the proteasome via a UBL domain that binds the proteasome and a UBA domain that binds to polyubiquitinated proteins. The importance of these shuttle proteins and the specific roles they play in proteasomal degradation are still being explored. To investigate the role of Dsk2 in proteasomal degradation, we examined degradation of model substrates by ∆Dsk2 proteasome in the presence or absence of purified Dsk2 protein. Surprisingly, we discovered that purified Dsk2 protein was able to remove ubiquitinated substrates from solution as determined using sedimentation assays. Dsk2’s ability to sediment substrates was dependent on ubiquitin chain linkage, with K63 linkages giving greater sedimentation than K48 or mixed linkages. Based on these findings, Dsk2‐induced substrate aggregation may play a role in the cellular fates of ubiquitinated proteins.
Lineage Tracking and Differentiation of Neuro 2A cells (N2a) upon exposure to Dopamine, Monosodium Glutamate and Corticosterone
Authors: Mathew Smita , Keerikkattil Joy P.
Published: May 2022

Cell to cell communication is critical for survival of an organism and it is achieved through signal transduction pathways. For the transmission of signal, molecules such as hormones can bind to a receptor on or inside the cell membrane. Dopamine (DA) a monoamine catecholamine neurotransmitter and hormone binds to dopamine receptor, inturn, affect movement, emotions and the reward system in the brain. The present study tested the hypothesis that Dopamine differentiates the N2a cells into neuronal lineage, and corticosterone (CORT) and monosodium glutamate (MSG) alter the differentiation in either way. Mouse neural crest‐derived cell line (Neuro2A cells, N2a cells) were treated with 10µM DA, MSG and CORT. Cells were seeded for different time points to track the lineage pattern of differentiation. Treatment was done for 24 hrs, 48 hrs, 72hrs and 96 hrs with respective control groups. Phase contrast microscopic images revealed that Dopamine treated cells showed neurite extensions and differentiation as time passed whereas CORT and MSG treatments suppressed the differentiation; the cells were round and showed signs of stress, cell death and autophagy. Immunocytochemical analysis with D1 receptor antibody indicated enhanced expression in the DA groups and diminished D1 receptor expression in CORT and MSG groups. Furthermore, quantitative analysis revealed that D1 receptor expression was significantly enhanced in DA group compared to the control group (medium containing ascorbic acid and phosphate buffered saline) and untreated control group (medium only). Interestingly D1 receptor expression in the DA group was significantly upregulated compared to the CORT group but not significantly different in the MSG group. Furthermore, inorder to assess apoptosis AnnexinV/Propidium Iodide (PI) staining was done using Fluorescence Assorted single cell sorter (FACS) Analysis. Immunolabeling with Caspase‐3 marker gene was done to detect apoptosis. N2a cells were treated with DA, CORT and MSG at 10 µM concentration for 24 hrs. Propidium iodide labelled cells that were already dead or in the late stage of apoptosis whereas Annexin V bound to the cells early in apoptosis and continued to be bound through cell death. Counting of the cells showed that MSG induced 1.8 % apoptosis, CORT induced 1.2 % and DA induced 1.1%, as against control group cells of 0.7 %. The total cell death including apoptosis and necrosis was highest in the MSG group (3.4%) compared to the control (1.7 %), DA (2.1%) and CORT (1.9%) treated groups. This data suggest differential effects elicited by DA (happy hormone) versus stress hormone. Our results also raise the question whether DA induces catecholaminergic pathway of differentiation, CORT stimulates glial type differentiation and MSG stimulates glutamate or GABA differentiation ‐ questions to be answered in future studies.
S‐acylation of STIM1 regulates store‐operated calcium entry
Authors: Kodakandla Goutham , West Savannah , Wang Qioachu , Tewari Ritika , Zhu Michael , Akimzhanov Askar , Boehning Darren F.
Published: May 2022

The initiating events happening during store‐operated calcium entry are not completely understood despite the tremendous importance of this process in immune responses. Upon endoplasmic reticulum (ER) calcium depletion, STIM1 and Orai1 form calcium release‐activated calcium (CRAC) channels to allow calcium influx from the extracellular milieu. S‐acylation is a reversible post‐translational modification of cysteine residues that affects protein structure, stability, and targeting to subdomains in the plasma membrane. Previously, we have shown that Orai1 is rapidly S‐acylated following T cell activation in Jurkat cells as well as ER calcium store depletion in HEK293 cells. Here we demonstrate that STIM1 is S‐acylated at C437 residue upon T cell activation and ER calcium store depletion. This is an interesting observation because very little is known about S‐acylation and its consequences in ER‐resident proteins. Upon store depletion, STIM1 adopts an extended conformation that permits binding to Orai1. We hypothesize that S‐acylation of STIM1 promotes and stabilizes the extended conformation of STIM1. Consistent with this hypothesis, we show that substitution of cysteine 437 in the STIM1 Orai1 activating region to a serine decreases calcium entry after store depletion. We further show that the ability of STIM1 to form puncta depends on its S‐acylation at this residue. Together with S‐acylation of Orai1, these data demonstrate that CRAC channel components are rapidly S‐acylated after ER calcium store depletion. In summary, based on our previous published literature on Orai1 S‐acylation, and current discovery of S‐acylation of STIM1, it is evident that S‐acylation recruits Orai1 to specialized membrane subdomains to form puncta with STIM1, whose extended conformation is stabilized by S‐acylation at its 437 residue. Hence, S‐acylation of Orai1 and STIM1 play a crucial role facilitate store‐operated calcium entry.
In Vivo Crosslinking and Mass Spectral Analysis of the Yeast Nucleosomal Protein Interactome
Authors: Fisher Raven
Published: May 2022

Chromatin is essential for proper DNA storage and gene regulation. The repeating unit of chromatin, the nucleosome, is composed of DNA wrapped around an octameric core of histone proteins. Histones interact with DNA, but they also make contacts to chromosomal regulating proteins. Histones undergo an array of dynamic posttranslational modifications (PTMs), each having an essential role in chromosomal structuring, function, and maintenance. To understand how nucleosomes regulate chromatin processes and how the dynamic PTM landscape influences histone‐protein interactions, it is important to identify interacting partners. We use UV crosslinking unnatural amino acids that allow for site‐selective photo‐crosslinking from histones, in vivo. This allows for the study of chromatin protein‐protein interactive dynamics in the physiological environment of the nucleus. We pair crosslinking techniques with mass spectral (MS) analysis to identify histone interacting proteins from cell lysates providing an unbiased overview of the site‐specific interactome. Our assays reveal many well‐established chromatin partners; however, they also provided insight to nucleosomal contacts that have been less reported in the literature. Of interest, we identify several histone contacts to ribosomal subunits, both large and small. We are interested in the extra ribosomal function of these proteins and their roles in chromatin regulation. To verify MS hits, we crosslink in yeast strains harboring a C‐terminal fusion tag (Myc) to the gene of interest, while expressing the crosslinking histone with a separate tag (HA). Products are precipitated and analyzed via western blot by decorating with antibodies to the opposing tag. We have verified several histone‐ribosomal protein interactions, including Rpl7A, Rpl3, and Rpl10. Using this technique, we investigate PTM mutations and their effects on the ability to form crosslinked products, providing insight on sequestering and regulatory mechanisms. These initial investigations suggest that UV crosslinking paired with MS analysis is an effective tool for studying yeast nucleosomal protein contacts.
SUMO 2 the rescue: how SUMO2 regulates the mitochondria via Drp1 modification
Authors: DeHaas Allison H. , Matunis Michael J.
Published: May 2022

Mitochondria are dynamic powerhouses of cells and their fission and fusion must be tightly regulated for normal cell function. Mitochondrial fission is mediated by the GTPase, Drp1, and mis‐regulation of Drp1 leads to mitochondrial hyper‐fragmentation, a known marker of disease. It has been reported that Drp1 is modified by SUMO1 and SUMO2/3, however, the mechanisms of Drp1 regulation by individual SUMO paralogs remain to be fully understood. Here, we have used CRISPR/Cas9 derived SUMO1 and SUMO2 knockout (KO) cell lines, to perform a systematic investigation of paralog‐specific effects on mitochondrial maintenance and function. In contrast to expectations, we observed multiple mitochondrial defects specifically in SUMO2 KO, but not SUMO1 KO, cells. SUMO2 KO cells had reduced mitochondrial activity based on results of MTT assays. By immunofluorescence microscopy, we also observed an increase in mitochondrial fragmentation in SUMO2 KO cells. Paradoxically, increased fragmentation occurred despite reduced levels of Drp1 protein expression and sequestration of Drp1 in large cytosolic foci. Taken together, our findings indicate that SUMO2 plays a non‐redundant, paralog‐specific role in regulating mitochondrial function, in part through effects on Drp1. We anticipate that a more detailed understanding of the molecular effects of SUMO2 on Drp1 may lead to novel therapeutic approaches to treat or prevent ischemic injury, neurodegeneration, and other mitochondrial‐related maladies.
Protease‐Activated Receptor‐2 Differentially Regulates Vascular Smooth Muscle Cell Proliferation in Cyclic AMP‐Dependent Protein Kinase/Phosphoinositide 3‐Kinase‐Dependent Manner
Authors: Williams Madison D. , Bullock Michael T. , Holland Nathan A. , Johnson Sean C. , Vuncannon Danielle M. , Oswald Joani Z. , Adderley Shaquria P. , Tulis David A.
Published: May 2022

Cardiovascular disease (CVD) remains the leading cause of morbidity and mortality worldwide, despite decades of extensive research. One key facet in CVD pathogenesis is the phenotypic transition of affected vascular smooth muscle (VSM) from a quiescent, contractile state to a synthetic, proliferative state. Protease‐activated receptors (PARs) are of emerging interest in CVD; however, their potential impact on VSM phenotypic switching inherent in CVD is unclear. The overall objective of the current study was to determine the role of PAR2 and PAR4 in phenotypic switching and VSM growth. We hypothesized that PAR2 and PAR4 increase following arterial injury in vivo and after growth stimulation in vitro, that they serve to protect against VSM cell growth, and that they signal primarily through specific protein kinases in phenotype‐specific fashion. In rat carotid arteries, PAR2 expression and activity increased following mechanical distension injury compared to uninjured controls. In rat primary VSM cells, PAR2 expression also increased in response to growth stimulation. Differential expression of cyclic GMP‐dependent protein kinase PKG‐1 and the PKG target VASP distinguished low passage (P3‐P6) from high P (P10‐P15) VSM cells, with low P cells showing elevated PKG‐1, phosphorylated (Ser239) VASP and total VASP compared to high P cells. Pharmacologic PAR2 agonism promoted DNA synthesis and proliferation in low P cells but decreased DNA synthesis and proliferation in high P cells. Western blotting analyses showed increased expression of MEK1/2 MAPK, PKA, and PI3K in low P cells versus high P cells. Specific protein kinase blockade revealed that PAR2 differentially controls proliferation of phenotypically distinct low versus high P VSM cells predominantly through PKA and PI3K. Our findings highlight potential (patho)physiological relevance of PAR2 and PAR2‐driven PKA/PI3K in the control of VSM cell growth and provide evidence for continued study of VSM PAR2 in CVD.
Intranasal Exposure to Hog Dust Extract Promotes Endotoxemia and Increases Gene Expression of TNF‐α and Secretary Epithelial Cell Markers in Mouse Intestine
Authors: Crawford Meli'sa S. , Ulu Arzu , Nordgren Tara M. , McCole Declan F.
Published: May 2022

Agriculture‐related dust exposures to antimicrobial‐resistant pathogens can have significant health implications for animal agricultural industry workers. In addition to major respiratory complications, air pollutants provoke intestinal diseases and altered digestive tract physiology. Hog farm dust is highly abundant in gram‐positive and gram‐negative bacteria. Lipopolysaccharides (LPS; endotoxins) excised from gram‐negative bacteria can drive increased intestinal permeability and inflammation. Therefore, we hypothesized that acute intranasal exposure to aqueous extracts of dusts collected from hog confinement facilities (HDE) ‐ containing bacterial LPS ‐ cause increased intestinal inflammation and barrier dysfunction. We previously showed that intranasal exposure of 8‐week‐old male or female C57BL/6J mice (n=12) to 12.5% HDE (containing 22.1‐91.1 EU/mL) (n=6) for 3 weeks elevated bronchoalveolar lavage total cell and neutrophil levels (indicative of chronic airway inflammation), and increased intestinal permeability, in comparison to saline controls (n=6). We now report that HDE‐treated mice have elevated serum LPS levels (unpaired t‐test, p<0.0001; n=6/group) suggesting endotoxemia. To further identify intestinal epithelial responses initiated by HDE, inflammatory genes were examined in intestinal whole tissue lysates by qPCR. TNF‐α expression was increased in the ileum (unpaired t‐test, p=0.0265; n=3‐5/group) but not in the colon of HDE‐treated mice. Conversely, IL‐1B, IL‐6 and IL‐10 were not significantly altered along the intestinal tract. Following HDE treatment, mRNA expression of the goblet cell marker, Muc2, was increased in distal colon epithelial cells (unpaired t‐test, p=0.0099, n=3‐5/group). The Paneth cell‐associated marker, Lyz1 (lysozyme), was unchanged in ileum but was increased in proximal colon epithelial cells (p=0.0444, n=2‐5/group) of HDE‐treated mice. Collectively, these data indicate that agricultural dust exposure induces endotoxemia and promotes intestinal inflammation characterized by increased TNF‐α and host anti‐microbial responses.
The Impact of Levels of Religious Belief and Practices on Heart Rate Variability
Authors: Paturzo Kathryn , Vick Robert S.
Published: May 2022

Heart rate variability (HRV) is the variation between consecutive heart beats over time. Increased HRV is positively correlated with better overall cardiac health. There are many lifestyle and biological factors that affect HRV. Research has examined the effects of chanting, reading holy texts, or praying on HRV, however, there has been little research done on how religion affects HRV. The purpose of this study is to determine whether there is a correlation between levels of religious beliefs and practice and HRV. Inter‐ and intra‐religious beliefs and levels of practice were measured using a scale from 1 to 5. HRVs were generated using an HRV analysis program (LabChart, ADInstruments) from a lead I electrocardiogram (EKG), measuring from one R wave to the next R wave (R‐R). EKGs, measuring 9 time‐domain measurements of HRV, were performed on 67 participants. T‐tests and Bonferroni analyses were performed on the data to determine significance. Data was analyzed from participants identifying as members of 4 different religious traditions: atheist, Catholic, non‐denominational Christian, and Jewish. There is a significant difference (p=0.03, n=10) in SDRR (std dev of R‐R time) of Catholics with different levels of religious practice, but not atheists (p=0.5, n=15), Christians (p=0.6, n=17), or Jews (p=0.7, n=11). Similarly, there is a significant difference (p=0.006, n=15) in SDRR of atheists with different levels of religious beliefs, but not Catholics (p=0.4,n=10), Christians (p=0.7, n=17), or Jews (p=0.2, n=11). The data supports significant differences in HRV between members of the same religious traditions having differing levels of practice, however the cause of this affect remains unknown.
A Role for Gβγ in Regulating Golgi fragmentation
Authors: Rajanala Kalpana , Gorsky Joshua , Wedegaertner Philip
Published: May 2022

Gβγ subunits regulate several non‐canonical functions at distinct intracellular locations. Previous work in our lab and others has established that Golgi‐localized Gβγ regulates a signaling pathway that controls the fission of protein cargo vesicles at the trans Golgi network (TGN). We recently demonstrated that Gβγ is required for mitotic Golgi fragmentation and established that knockdown or inhibition of Gβγ results in delayed mitotic progression. This study is focused on understanding the role of Gβγ in regulating Golgi fragmentation resulting from disruption of microtubules which occurs in response to drugs and disease states. We used siRNA against Gβ or specific Gγ subunits to deplete their expression, and demonstrated that their knockdown causes a significant decrease in nocodazole‐induced Golgi fragmentation. In addition, our results show that knockdown or inhibition Gβγ causes decreased activation of protein kinase D (PKD) in response to nocodazole treatment. These results implicate a role for Gβγ in regulating Golgi structure, and future experiments will help us to understand the signaling mechanisms regulated by the Gβγ subunits at the Golgi.
Role of a conserved family of serine hydrolases in ethanol toxicity
Authors: Leaman Megan , Barton Jennifer , Johnson R. Jeremy
Published: May 2022

A recent CRISPR screen for genes involved in controlling acetaldehyde dependent ethanol toxicity, identified OVCA2, a conserved serine hydrolase, as the highest protectant against acetaldehyde toxicity. OVCA2 is a member of a larger, diverse group of proteins responsible for metabolic activities known as the family of serine hydrolases (FSH). In S. cerevisiae,three FSH homologues (FSH1, FSH2, and FSH3) are present, but their overlapping functions and their connection to acetaldehyde toxicity are unknown. In this study, we worked to understand the role of FSH proteins in controlling acetaldehyde toxicity in two divergent ways. First, we compared the substrate specificity of the three FSH homologues from S. cerevisiae against a library of ester substrates at varying pH environments. Second, we constructed combinatorial deletions of FSH genes in S. cerevisiae and screened deletion strains for changes in growth under high ethanol, acetaldehyde, and varying other toxic aldehydes. For enzyme activity, both FSH2 and FSH3 had a preference for alkaline conditions and thus the comparative substrate specificity screen was completed at pH 7.5. In this screen, FSH2 and FSH3 showed substrate preferences in line with FSH1, as they both preferred short chain esters versus longer chain esters (>8C). To further evaluate the metabolic activity of FSH enzymes in acetaldehyde toxicity, single, double, and triple deletion strains of the three FSH genes were made in S. cerevisiae. Proper gene deletions for each strain were confirmed by colony PCR. Each of the deletion strains were then grown on agar plates with varying concentrations of alcohol and aldehyde toxins. Each of the single deletion mutants showed similar growth rates to each other and to the wild‐type strain by spot dilution but exhibited slightly different colony morphology from the wild‐type strain. Further testing on the ability to withstand toxins produced by the alcohol metabolism pathway will be conducted to determine whether these enzymes and OVCA2 can metabolize the toxic byproducts of ethanol production.
Investigating the Effect of Small Extracellular Vesicles at Varying Concentrations on Wound Healing
Authors: Henretta Sara , Kasper Anna , Gaborski Thomas
Published: May 2022

Extracellular vesicles (EVs) are nanoscale lipid bilayer particles that are secreted by cells into their surrounding environment. EVs in the extracellular space taken up by nearby cells can influence the recipient cells’ phenotype. For this reason, EVs are believed by some to influence cell‐cell communication, specifically cell motility. While previous literature has claimed EVs have a significant effect of wound healing and cell recovery, we believe these results are due to cells being dosed in super supraphysiological levels of EVs. We hypothesize that cells dosed with EVs at physiological levels will have a less significant effect on wound healing properties than what has previously been seen. This study investigates the effect of adipose derived stem cell (ADSC) derived EVs on human dermal fibroblast (hDFb) migration and wound healing speeds using time lapse imaging and cell migration analysis code.

Cell number recovery (CNR) is the ratio of the number of cells which have entered the wound region to the initial number of cells in the wound region prior to scratching. Initial number of cells in the wound region was estimated from the starting density of unscratched cells outside the wound region. A CNR of 1 indicates the wound has fully recovered and reached its initial density. We compared CNR in media control (complete media) and baseline control (serum free media) to cell recovery in EV dosed media with a concentration of 5*108 EVs/mL. EV doses were calculated such that they corresponded to EVs produced by ADSCs within 24 hours at a ratio ranging from 1:1 (5*109 EVs/mL) to 1:10 (5*108 EVs/mL) ADSCs to hDFbs. These ratios were chosen as they likely mimic the upper limits of physiological ratios during wound healing. Previous studies often used very high levels of EVs that may be relevant for therapeutic approaches. A vascular endothelial growth factor (VEGF) ELISA was performed to confirm growth factors did not co‐precipitate during isolation and did not influence the scratch wound assay results. EV Depleted media, collected from the supernatant of the final ultracentrifugation spin, confirmed the presence of VEGF. Conversely, both serum free and small EVs measured below the limit of detection for VEGF. These results confirm the slight increase in CNR is not due to growth factors that co‐isolated with EVs. In conclusion, EVs may provide a therapeutic effect for wound healing when dosed at supraphysiological levels, however, at more physiological levels, only a minor difference in wound healing is seen between the EV dosed cells and baseline controls.
The Effects of Point Mutations on the Dimerization Domain of Ebola Virus Protein VP40
Authors: Conarty Jacob , Patel Himani , Wiest Olaf , Amiar Souad , Stahelin Robert
Published: May 2022

The Ebola Virus (EBOV) is a deadly virus first identified in 1976 from the Ebola River region of Zaire (presently the Democratic Republic of Congo). EBOV is known for causing viral hemorrhagic fever and it can cause a high mortality rate in infected patients. VP40 is the matrix protein of EBOV (eVP40), which has an N‐terminal domain responsible for dimerization and oligomerization of the protein to form filaments of dimers at the plasma membrane inner leaflet of the host cell. The eVP40 dimerization and filament formation is necessary for viral budding from the host cell membrane. This project aims to investigate which amino acid residues in this N‐terminal dimerization domain are necessary for the protein to be able to oligomerize at the plasma membrane inner leaflet. First, computational screens were performed to determine which residues may play a role in dimerization, and then mutant plasmids with changes at these residues harboring a green fluorescent protein tag were created. Based on the computational screen, we hypothesized amino acid residues Arg52, Ile54, Ala55, Asp56, Asp57, and His61 would be essential for dimerization of eVP40. Mutants of these residues were then transfected into Cos‐7 cells and imaged using confocal microscopy and compared to WT eVP40. Cellular images were analyzed using ImageJ to determine the amount of fluorescently‐tagged protein present at the plasma membrane. Confocal imaging confirmed our central hypothesis and the computational data as these mutations had a significant loss of eVP40 plasma membrane localization. The next steps in this project will be to perform virus‐like particle (VLP) collection to further analyze the effects of point mutations on the dimerization domain as it relates to VLP formation budding out of the host cell.
The Effect of Meditation on Heart Rate Variability
Authors: Gonzales Rhianna , Hartman Sarah R. , Luckenbill Courtney , Logan Hannah , Cassel Spencer , Kieffer H. S.
Published: May 2022

Heart rate variability (HRV) has been utilized in both medicine and sports sciences as an important marker for managing stress, indicating diseased states, and monitoring training/recovery. HRV reflects the autonomic balance of the sympathetic (SNS) and parasympathetic (PNS) nervous systems. High PNS activity is correlated to high HRV. The frequency and time domains of an ECG may be analyzed to determine the influence of PNS activity. HRV has been shown to be influenced by exercise training; however, meditation and guiding breathing techniques may also influence HRV.

PURPOSE

To evaluate the effect of a short, guided meditation session on the measures of heart rate variability of inexperienced meditators.

METHODS

39 participants (18‐21 years) volunteered and gave informed consent. During the experimental session, a Polar H10 heart rate monitor was fitted around the subject to record a 15‐minute ECG. The breathing protocol consisted of sitting in a comfortable position and continuous 5‐minute stages of spontaneous breathing (SP1), controlled breathing (CB), and returning to spontaneous breathing (SP2). During CB, a 6 breath/min guided protocol was followed using a commercially available App. A one‐way ANOVA with repeated measures was conducted for each variable using SPSS, p > 0.05.

RESULTS

CB produced a significant increase in heart rate from 70.6+11.9 bpm to 72.9+10.4 bpm (p<0.005) during CB, and then decreased to 71.7+10.3 bpm (p<0.05). Measures of the frequency domain included high frequency (HF) and low frequency (LF) power. HF significantly decreased from 49.2+21.1 nu during SP1, to 16.6+9.9 nu (p<0.005), and returned to 34.6+18.3 nu (p<0.005). For LF, SP1 was 50.9+21.0 nu, significancy increased to 83.4+9.9 nu (p<0.005), and returned to 65.4+18.3 nu (p<0.005). The measures of the time domain included rMSSD and SDNN. rMSSD at SP1 was 60.9+33.8, significantly increased to 74.2+36.6 (p<0.005), and returned to 56.0+33.1 (p<0.005). The SP1 for SDNN was 62.7+31.4, significantly increased to 110.4+39.6 (p<0.005), and returned to 66.1+29.7 (p<0.005). CONCLUSION: The direction and magnitude of change in frequency domain (HR, HF and LF) may initially suggest a dampening of the PNS. However, CB has been shown to initiate SNS activity which is represented in the frequency measures. In short‐term HRV measures, rMSSD and SDNN are the more appropriate measures to consider for the PNS activity. Thus, the results of this study indicate that a short session of CB may stimulate a PNS response.
Faster skin wound healing predicts survival after myocardial infarction
Authors: Becirovic‐Agic Mediha , Chalise Upendra , Jung Mira , Rodriguez‐Paar Jocelyn R. , Konfrst Shelby R. , Flynn Elizabeth R. , Salomon Jeffrey D. , Hall Michael E. , Lindsey Merry L.
Published: May 2022

Skin wound healing and the cardiac healing response to myocardial infarction (MI) both progress through similar pathways involving inflammation, resolution, tissue repair, and scar formation. Due to the similarities, we hypothesized that the healing response to skin wounding would predict future response to MI. Mice were given a 3 mm skin wound using a disposable biopsy punch, and the skin wound was imaged daily until closure. Wound closure was quantified using Image J software. The same set of mice was given MI by permanent coronary artery ligation 28 days later and followed for 7 days. Cardiac physiology (volume, dimension, ejection fraction, fractional shortening, wall thickness) was measured by echocardiography at baseline, and 3 and 7 days post MI. Mice that survived until MI day 7 were grouped as survivors (30 mice, 68%), and animals that died from MI were grouped as non‐survivors (14 mice, 32%). Survivors had faster skin wound healing compared to non‐survivors. Faster skin wound healing predicted MI survival better than commonly used cardiac functional variables (e.g., infarct size, fractional shortening, and end diastolic dimension). We mapped the plasma N‐glycoproteome 3 days after skin wounding and 3 days after MI using mass spectrometry. The glycoproteome profile of MI day 3 plasma revealed alpha‐2‐macroglobulin and ELL‐associated factor 1 as strong predictors of future MI death and progression to heart failure. A second cohort of MI mice validated these findings. In patients, alpha‐2‐macroglobulin was 1.7±0.3‐fold elevated 48h after presentation of MI (p=0.009, 18 controls, 41 MI patients). The glycoproteome profile of plasma collected 3 days post skin wounding identified apolipoprotein D and vitamin D binding protein to mirror skin wound healing. Apolipoprotein D detrimentally regulated both skin and cardiac wound healing in part by promoting inflammation. Plasma levels of vitamin D binding protein and galectin 3 binding after skin wounding also predicted future MI dilation and progression to heart failure. Our results reveal that the skin is a mirror to the heart and common pathways link wound healing across organs.
Can Finnish Sauna Bathing Induce Heat Acclimation in Adults with Stable Coronary Artery Disease?
Authors: Bartlett Audrey‐Ann , Garceau Laurence , Gravel Hugo , Debray Amélie , Nigam Anil , Gagnon Daniel
Published: May 2022

Introduction

Extreme heat is associated with a greater risk of mortality that particularly affects older adults with cardiovascular disease (CVD). Regular heat exposure induces an adaptation of thermoregulatory mechanisms (acclimation) that may alleviate thermal and/or cardiac strain during subsequent heat exposure. However, limited research has examined practical heat acclimation protocols that could be recommended to vulnerable populations, such as adults with CVD. The objective of this study was to test the hypothesis that 8 weeks of Finnish sauna bathing reduces core temperature and heart rate and increases sweating during heat exposure.

Method

14 participants (11 men/3 women, 63 ± 5 years, 28.7 ± 3.8 kg/m2) with stable coronary artery disease participated in an 8‐week Finnish sauna intervention (4 sessions/week, 20 to 30 min/session, at ~80°C, 20% humidity). Core temperature (ingestible telemetric pill), heart rate (Polar watch), and sweat rate (change in nude body weight) were measured during the first and last sauna sessions.

Results

Resting core temperature prior to the first sauna session was 37.4 ± 0.4°C, compared to 37.1 ± 0.3°C prior to the last session (p= 0.04). Core temperature at the end of sauna bathing did not differ between the first (37.77 ±0.34°C) and the last (37.59 ± 0.31°C, p=0.20) sessions. Heart rate at the end of sauna bathing was greater during the last (90 ± 17 bpm) compared to first (85 ± 16 bpm, p=0.13) sessions. Sweat rate was greater during the last sauna session (first session: 8.6 ± 5.9 mL/min vs. last session: 5.1 ± 3.0 mL/min, p<0.01).

Conclusion

Eight weeks of Finnish sauna bathing reduces resting core temperature and increases sweat rate during heat exposure in adults with stable coronary artery disease. However, these adaptations did not translate into reduced thermal and cardiac strain during heat exposure.
Investigating the Effect of PARP Mediated Compartments on Transcription Dynamics
Authors: Eng Vincent F. , Versluis Philip , Lis John T.
Published: May 2022

Transcription of DNA into RNA is a fundamental, universal, and highly regulated process in all living organisms. RNA Polymerase II (Pol II), is the enzyme that catalyzes this reaction and it is highly regulated by the pausing factors DRB Sensitivity Inducing Factor (DSIF) and Negative Elongation Factor (NELF), and elongation factors, including Positive Transcription Elongation Factor b (P‐TEFb) and Polymerase Associated Factor 1 (PAF1). However, most previous studies of these factors have only provided snapshots of factor binding or snapshots of mRNA production following their depletion. While valuable, these studies provide limited information about the time frames in which Pol II and key transcription factors interact with each other and with the DNA. Using live‐cell imaging of amplified and interphase‐extended Drosophila polytene chromosomes with endogenously tagged transcription factors and Pol II, we can analyze the real time behavior of these molecules at single genetic loci and at high resolution. In particular, heat shock genes, like heat shock protein 70 (Hsp70), have been shown to be a robust system for studying transcription because of their inducible nature; they are able to transition quickly from a paused to a highly active state. Previous work from our lab utilizing this system suggested the formation of a transcription compartment during prolonged bouts of transcription. We propose that transcription compartments form around Hsp70 loci during an extended heat shock due to the activity of Poly ADP Ribose Polymerase (PARP) polymerizing large PAR chains that trap Pol II and other transcription factors, restricting their movement and preventing free diffusion with other molecules in the nucleus. We are tagging proteins of interest with photo‐activatable fluorescent proteins to minimize background by selectively activating a population of molecules only in the small 3‐D volume of specific loci. Utilizing this system we have begun measuring the dynamics of Pol II at specific loci and are working toward measuring the dynamics of key transcription regulating factors and further characterizing PARP‐mediated transcription compartments.