bluuebunny/crossref_metadata_2025 huggingface dataset:
DPO‐1‐sensitive K+ channels control the tone and reactivity of resistance‐sized arterioles from brain and skeletal muscle (678.5)
Authors: Butcher Joshua , Fancher Ibra , Frisbee Jefferson , Dick Gregory
Published: June 2021

Arterial smooth muscle cells possess prominent delayed rectifier K+ (KDR) currents. KDR currents may be mediated by KV1.5 channels, which are sensitive to inhibition by DPO‐1. We tested the hypothesis that DPO‐1‐sensitive KDR channels control the tone and reactivity of resistance‐sized arterioles from brain and skeletal muscle. Rat (n = 4) middle cerebral and gracilis arterioles were dissected, cannulated, and pressurized to 80% of mean arterial pressure (86 ± 3 mmHg) for video microscopy measurements. With active tone, the diameters of gracilis and cerebral arterioles were 107 ± 5 and 103 ± 4 μm. DPO‐1 (10 μM) constricted gracilis and cerebral arterioles to 79 ± 4 and 73 ± 5 μm. DPO‐1 increased the sensitivity (logEC50) to phenylephrine (6.3 ± 0.1 vs 6.9 ± 0.1) and serotonin (6.5 ± 0.1 vs 7.2 ± 0.1) in gracilis and cerebral arterioles, respectively. Increasing internal pressure from 40 to 140 mmHg in 20 mmHg steps activated myogenic responses (diameter decreased 0.18 ± 0.05 and 0.17 ± 0.05 μm/mmHg in gracilis and cerebral arterioles). DPO‐1 eliminated myogenic responsiveness, as diameter increased 0.08 ± 0.07 and 0.06 ± 0.05 μm/mmHg. Dilation to 100 μM acetylcholine was 27 ± 3 and 27 ± 1 μm in gracilis and cerebral arterioles; DPO‐1 reduced these dilations to 4 ± 2 and 10 ± 2 μm. We suggest that DPO‐1‐sensitive KDR channels (KV1.5) play a major role in determining microvascular tone and responses to vasoconstrictors and vasodilators.

Grant Funding Source: Supported by T32HL090610
Relief of endoplasmic reticulum stress in the brain rescues high fat diet‐induced hypertension but this is not due to the subfornical organ (682.3)
Authors: Young Colin , Butler Scott , Horwath Julie , Mark Allyn , Davisson Robin
Published: June 2021

We have recently demonstrated robust ER stress in central neural cardioregulatory regions of HFD animals, with particularly prominent elevations in the SFO. In line with this, global relief of brain ER stress via intracerebroventricular delivery of the chemical ER chaperone TUDCA ameliorates HFD‐induced hypertension (Δ‐6±1 mmHg, p<0.05). However, the precise neural regions involved remain unknown. We hypothesized that ER stress in the SFO mediates obesity‐induced hypertension. HFD (n=13; 20 wks) and age‐matched normal chow (n=10) fed C57Bl/6 mice were instrumented with radiotelemeters for mean arterial pressure (MAP) measurements and underwent SFO‐targeted microinjections of an adenovirus encoding the ER chaperone GRP78 (AdGRP78) to reduce ER stress. Relative to normal chow, HFD control vector treated mice demonstrated a significant increase in MAP (102±3 vs 116±1 mmHg, normal chow vs HFD, p<0.05). SFO‐targeted AdGRP78 did not influence the HFD‐induced increase in MAP (118±1 mmHg, HFD AdGRP78, p>0.05). Analysis of ER stress biomarkers revealed that AdGRP78 selectively reduced ER stress within the SFO, but not other neural regions (e.g. p58IPK mRNA SFO: 1.0±0.1 vs 0.6±0.1, HFD AdLacZ vs HFD AdGRP78, p<0.05). These findings suggest that, while brain ER stress contributes to HFD‐induced hypertension, ER stress in the SFO does not appear to be involved in the maintenance of obesity‐mediated hypertension.

Grant Funding Source: Supported by HL63887, HL96571, HL84207, AHA13POST14410020, K99HL166776
Cordyceps sinensis Cs‐5 improves memory and learning abilities in a dysmnesia model (684.1)
Authors: Dong Yunzi , Yang Jieying , Zhang Yan , Zhao Chunsheng , Wu Zimei , Tan Ningzhi , Lu Jihong , Bartlett Mark , Zhu Jia‐shi
Published: June 2021

Cordyceps sinensis Cs‐5, a mycelia fermentation product of Cordyceps sinensis (Cs), has shown its health benefits in modulating immune functions and enhancing sexual functions. This study was to examine the effects of Cs‐5 on improving memory and learning abilities in a mouse model with memory impairment. ICR male mice (2 months) were randomized into 3 groups on the basis of the pre‐study swimming latency in water maze test, and received Cs‐5 at a dose of 0.5 or 1.5 g/kg, or vehicle for 60 days. The mice received intraperitoneal injection of Scopolamine Hydrobromide at a dose of 5mg/kg for memory acquisition impairment, and were tested with water maze and step‐through tests to evaluate the memory and learning abilities. Compared to controls, mice received Cs‐5 at a dose of 0.5 g/kg (equivalent to the human recommended dose) showed significantly shortened swimming latency in water maze test, and significantly prolonged step‐through latency in step‐through test (both p=0.03). In conclusion, our results indicate that Cs‐5 is effective on improvement of memory and learning abilities in a mouse model with memory impairment, supporting its potential anti‐aging effect.
Dynamic regulation of microRNA networks in the brainstem underlie hypertension development (686.22)
Authors: DeCicco Danielle , Zhu Haisun , Brureau Anthony , Schwaber James , Vadigepalli Rajanikanth
Published: June 2021

This study aims to elucidate the mechanisms by which microRNAs (miRNA) regulate autonomic control networks driving hypertension development. Using a high‐throughput assay for 419 miRNAs in the Spontaneously Hypertensive Rat (SHR) and Wistar‐Kyoto (WKY) control at three key stages of hypertension development, significant differences appear in 23 miRNAs of the nucleus tractus solitarius (NTS) and rostral ventrolateral medulla (RVLM). Alterations were stage‐ and region‐dependent, with significant differences occurring at mid‐stage in NTS vs early‐stage in RVLM. 96 gene targets corresponding to catecholaminergic processes and neuromodulation were predicted using the miRWALK database and tested using high‐throughput qPCR. Using correlational relationships between miRNAs and mRNAs, we observed a double‐negative regulatory motif, which appears prevalent throughout the data. This motif consists of a miRNA down‐regulating a negative regulator of a pro‐hypertensive signaling pathway like Angiontensin II signaling or leukotriene‐based inflammation. We demonstrated for the first time that the broad concordance of microRNA dynamics and target gene expression compose a regulatory network in the brainstem underlying hypertension.

Grant Funding Source: Supported by NIGMS R01 GM083108, NHLBI R01 HL111621
Central projections of sodium‐sensitive afferent renal nerves in rats (687.3)
Authors: Goodwill Vanessa , Loewy Arthur , Knuepfer Mark
Published: June 2021

Renal denervation is reported to treat resistant essential hypertension. Many suggest that afferent renal nerves (ARN) are responsible for the full expression of hypertension. We hypothesized that osmosensitive ARN activate sites in the CNS that contribute to hypertension by evoking excessive sympathetic nerve activity. To determine the central projections, a dual lumen cannula was implanted through the left ureter into the renal pelvis of the rat. Several days later, renal pelvic afferents were bathed with different concentrations of NaCl (20, 308 and 500 mEq/L) using a push‐pull perfusion for 2 hrs while recording arterial pressure in conscious rats. Activation of CNS sites was identified using immunohistochemical detection of c‐Fos protein and compared to controls (no infusion or denervation). Many sites were dose‐dependently stimulated in response to intrarenal infusion of NaCl. These included the nucleus tractus solitarius, area postrema, parabrachial nucleus, paraventricular hypothalamus, and other areas. The results indicate that stimulation of osmosensitive ARN leads to activation of central autonomic sites involved in the regulation of sympathetic outflow. Thus, this work identifies likely sites mediating ARN‐induced sympathoexcitation contributing to the pathogenesis of hypertension.

Grant Funding Source: Supported by USPHS HL25449 (ADL) and DA017371 (MMK).
SMAD1 signaling mediates inhibition of store‐operated Ca2+ entry on collagen IV expression in glomerular mesangial cells (689.14)
Authors: Wu Peiwen , Wang Yanxia , Chaudhari Sarika , Zuckerman Jonathan , Davis Mark , Ma Rong
Published: June 2021

Objective: The Smad signaling pathway plays a critical role in extracellular matrix protein production in glomeruli in diabetic nephropathy. Store‐operated Ca2+ entry (SOCE) regulates protein synthesis. The aim of the present study was to determine if SOCE in mesangial cells (MCs) suppressed production of collagen type IV (Col IV) by inhibiting Smad1.

Methods: Western blot and immunohistochemistry were used to detect target protein expression in cultured MCs and mouse glomeruli, respectively. Systemic administration of nanoparticles containing siRNA against Orai1 was employed to deliver the siRNA to MCs in mice.

Results: In cultured human MCs, activation of SOCE by cyclopiazonic acid or thapsigargin (TG) significantly suppressed Ang II‐induced Smad1 phosphorylation. Overexpression of STIM1 dramatically decreased abundance of Smad1 protein. Furthermore, high glucose‐induced Col IV protein expression was significantly inhibited by TG. Inhibition of SOCE by 2‐aminoethoxydiphenyl borate or La3+ increased Col IV expression which was accompanied by an increase in Smad1 protein expression. However, the Col IV response to SOCE inhibition was not observed in MCs with knockdown of Smad1. Moreover, in vivo knockdown of Orai1 in mouse MCs increased expressions of both Smad1 and Col IV in the nanoparticle‐positive glomeruli.

Conclusion: SOCE inhibits Col IV expression by inhibiting Smad1 pathway in MCs.

Grant Funding Source: NIH/NIDDK
Multilevel synchronization of tubuloglomerular feedback is modulated by nitric oxide (692.10)
Authors: Mitrou Nick , Scully Christopher , Braam Branko , Chon Ki , Cupples William
Published: June 2021

Previously we showed that NO modulates synchronization of both mechanisms of renal autoregulation. We did not, however, provide a mechanism to explain TGF synchronization after L‐NAME. L‐NAME changed TGF synchronization in a way we did not understand. Here we show that TGF synchronization may become stronger within cortical lobules after L‐NAME, resulting in two mechanisms of synchronization within a synchronized cluster. Male Long‐Evans rats (N=8) were anaesthetized (isoflurane) and their left kidneys were exposed. Cortical perfusion during control and NOS inhibition (L‐NAME, 10mg/kg i.v.) was monitored by laser speckle perfusion imaging. The TGF frequency range (0.01‐0.06 Hz) was isolated with a band pass filter. Clusters of synchronized nephrons were identified based on phase coherence (PC) between all pairs of imaged pixels. During control, TGF was synchronized moderately over large areas (~5x7 mm). After L‐NAME, each cluster had areas with high PC (strong synchronization) in the center and lower, but significant, PC in the surround. When the PC reference pixel was moved systematically there was strong synchronization within smaller ~800 x 800 um areas. The position of the high PC areas was consistent with an anatomical basis for synchronization. We conclude that TGF synchronization becomes stronger within cortical lobules while remaining weakly synchronized among lobules within the larger clusters.

Grant Funding Source: Supported by CIHR MOP‐102694
Biotin ligase tagging identifies E‐cadherin protein neighbors, including lipoma preferred partner, which modulates epithelial cell‐cell and cell‐substrate adhesion (694.4)
Authors: Van Itallie Christina , Tietgens Amber , Aponte Angel , Fredriksson Karin , Fanning Alan , Gucek Marjan , Anderson James
Published: June 2021

Proteins known to be associated with cellular adherens junctions (AJs) include E‐cadherin, catenins and other proteins involved in actin organization, vesicular trafficking and various signaling pathways. However, it is likely that some proteins with important roles at the AJ have not yet been identified. To detect additional AJ proteins, we expressed E‐cadherin fused to an engineered biotin ligase in MDCK epithelial cells and purified proximal, biotinylated proteins; these were identified by mass spectrometry. Catenins were among the most abundant of the 300+ proteins identified, along with nearly 40 other proteins previously reported to influence the AJ functions. Other proteins could be easily understood as novel candidates for regulating the structure or function of the AJ. One protein, lipoma preferred partner (LPP) was chosen for further characterization. Knockdown of LPP in MDCK cells demonstrated it was required for normal E‐cadherin‐dependent adhesion and was likely involved in coordination of cell‐cell and cell‐substrate adhesion. The demonstration of a role for LPP in E‐cadherin‐dependent adhesion was a proof of principle that the analysis of E‐cadherin protein neighbors could generate new tools to aid in AJ investigation.

Grant Funding Source: Supported by the Intramural Research Program of the NIH.
N‐cadherin adhesion mediated signaling stabilizes VE‐cadherin‐mediated adhesion and strengthens endothelial barrier function (695.3)
Authors: Kruse Kevin , Sieracki Nathan , Komarova Yulia
Published: June 2021

N‐cadherin adhesion mediated signaling stabilizes VE‐cadherin‐mediated adhesion and strengthens endothelial barrier function Kevin Kruse, Nathan Sieracki, Yulia Komarova.

Neuronal (N)‐cadherin has been shown to be involved in maintaining endothelial barrier function by forming homophilic interaction between endothelial cells (ECs) and pericytes, although it is unknown whether this is solely due to recruitment of pericytes to the endothelium or whether there are signaling events downstream of N‐cadherin homophilic adhesions in ECs that may play a role in strengthening the endothelial barrier. Our research objective therefore is to understand the role of N‐cadherin adhesion mediated signaling in regulating endothelial barrier function. Because N‐cadherin does not form homotypic interactions in an in vitro cell culture monolayer, we have utilized surface chemistry techniques to mimic the N‐cadherin interactions found between ECs and pericytes in vivo. The extracellular domain of N‐cadherin was covalently attached to an Ni‐NTA cover slip, allowing us to study N‐cadherin‐mediated adhesion in ECs. Our data show that ECs grown on N‐cadherin coated surfaces have increased VE‐cadherin accumulation as well as decreased t½ (half‐time to full fluorescence recovery) of VE‐cadherin turnover at cell‐cell junctions as determined by immunofluorescent staining and fluorescent recovery after photobleaching (FRAP). Our data indicate that N‐cadherin outside‐in signaling promotes formation of VE‐cadherin‐mediated adhesion by possibly stabilizing VE‐cadherin trans‐interactions. We will further use VE‐ or N‐cadherin biomimetic surfaces to isolate either VE‐cadherin or N‐cadherin complexes in order to understand the common and distinct binding partners and signaling pathways downstream of VE‐ and N‐cadherin.

Grant Funding Source: Supported by NIH T32
Experience for using platelet‐rich plasma in the treatment of androgenic alopecia (695.8)
Authors: Batpenova Gulnar , Sadykova Gulnaz , Kotlyarova Tatiyana
Published: June 2021

One of new physiological and correspondingly safe treatment in dermatology is PRP (PRP). Literature data suggest a positive PRP role in hair growth.

Objective: to evaluate the efficacy and safety of monotherapy PRP in the treatment of androgenetic alopecia.

Material and Methods: 15 patients with androgenetic alopecia , including male ‐9, 6 ‐ women aged 21 to 58 years , average age ‐ 40.2 years. Duration of disease, on the average is 6.5 years. Monotherapy was carried out by intradermal injection of platelet rich plasma at intervals of 2 months. The number of procedures is 1‐4, on the average ‐ 2. Evaluation of the results was made by macrofotografic way. The observation period is 12 months.

Results: as a result of platelet‐rich plasma therapy, there is a positive trend established as the form of hair loss reduction and enhancing their growth. Patients receiving more than 3 treatments achieved the significant effect. During and after treatment the side effects for a year have not been noted.

Conclusions: The results of the treatment are definitely confirmed by the macrophotografic data. However, the present study requires a careful study the influence of platelet rich plasma, not only to the growth of hair, but also on the structure, which suggests to conduct more extensive controlled studies using methodics such as dermoscoy, video dermoscopy, trihoscopy, determination Ki‐67 markers in a large cohort of patients.
Elevated carboxyhemoglobin in a marine mammal (698.1)
Authors: Tift Michael , Ponganis Paul , Crocker Daniel
Published: June 2021

Low concentrations of endogenous carbon monoxide (CO), generated through catabolism of heme‐proteins, have been shown to protect tissues against inflammatory diseases, and ischemia‐reperfusion injuries. However, elevated carboxyhemoglobin (COHb) formed by CO binding to hemoglobin, prevents adequate O2 delivery to tissues by lowering arterial O2 content. Therefore there must be a balance in the level of endogenous CO which maximizes cytoprotective effects and minimizes inhibition of O2 delivery. The elevated heme‐protein concentrations as found in marine mammals, are likely associated with increased heme catabolism, and subsequently increased endogenous CO production and COHb values. We measured COHb in elephant seals, a species with exceptional blood volumes, hemoglobin concentrations and myoglobin content. The mean value in adult seals was 8.7 ± 0.3%, while juveniles and pups (with lower heme‐protein stores) had COHb values of 7.6 ± 0.2% and 7.1 ± 0.3%, respectively. Serial samples over several hours revealed little to no fluctuation in COHb values. This consistent elevation in COHb suggests the magnitude and/or rate of heme‐protein turnover, is higher than terrestrial mammals. While these COHb values decrease the O2 stores needed for diving, the constant presence of elevated CO in blood may provide protection against ischemia‐reperfusion events associated with breath‐holds in elephant seals.

Grant Funding Source: Supported by National Institute of Health NHLBI RO1‐HL091767
What is the role of S‐nitrosation of soluble guanylyl cyclase in the heart? (698.9)
Authors: Huang Can , Crassous Pierre‐Antoine , Beuve Annie
Published: June 2021

Nitric oxide (NO) is involved in cardioprotection against ischemia/reperfusion (I/R) injury. Soluble guanylyl cyclase (sGC) produces cGMP when it is stimulated by NO. Association of the α and β subunit of sGC is required for cGMP‐forming activity. NO protects the heart from I/R via a cGMP‐dependent and a S‐nitrosation (SNO)‐dependent pathway. Preliminary evidence showed that sGC‐overexpression increases S‐nitrosation of specific proteins (SNO‐proteins) in rat neonatal cardiomyocytes. These sGC‐dependent SNO‐proteins were identified by Mass Spectrometry and are mostly associated with intercalated disc and cytoskeleton, including integrin‐linked kinase (ILK) and desmin. sGC is heavily S‐nitrosated in the presence of nitrosating agent. Thus, we hypothesize that sGC acts as a transnitrosylase in the heart, e.g. transfers its SNO to specific proteins. Intriguingly, sGC‐α and β subunits localize differently in cardiomyocytes. β is at the nuclear envelope and sarcolemma, while α is in cytosol, suggesting that each subunit might have a separate function, independent of cGMP‐forming activity. In cardiomyocytes treated with a nitrosating agent, overexpressing a single sGC subunit, either α or β, was sufficient to increase SNO level of ILK and desmin. Hence, we propose that sGC could modulate cardioprotection by increasing protein‐SNO, in addition to the NO‐cGMP dependent pathway.

Grant Funding Source: Supported by 6R01 GM067640
Genetic deletion of Kvβ1.1 attenuates isoproterenol induced cardiac injury (699.1)
Authors: Tur Jared , Chapalamadugu Kalyan , Panguluri Siva , Tipparaju Srinivas
Published: June 2021

The physiological roles of voltage gated potassium channel subunit (Kvβ1.1) remain unknown. The β‐subunits are highly expressed in the heart and bind Kv1.x and Kv4.x channel. We hypothesize that the Kvβ1.1 plays significant roles in sensing and altering the cardiac function when subjected to isoproterenol (Iso) induced injury. Mice were exposed to saline or Iso infusion for 14 days in age matched Wild type (WT) and Kvβ1.1 knockout (KO) groups. Echocardiography data indicates that Iso induced cardiac hypertrophy in WT mice along with a significant drop in ejection fraction (64±1.6 vs. 52.7±1.6), fractional shortening (34.4±1.9 vs. 26.9±1) (p蠄0.05), and dilated LVID(d), LVID(s), however, no significant differences were observed in KO groups. Monophasic action potential (MAP’s) recordings were unchanged in the Iso‐KO mice, whereas the APD’s were prolonged in the Iso‐WT. The electrophysiological data combined with Echo data clearly reveal attenuation in Iso‐KO indicating protection. Serum cardiac troponin I and LDH levels were increased in Iso‐WT whereas no increase was noted in KO groups. Western blot analysis revealed a decrease in Kv4.2 in WT‐Iso mice whereas KO‐Iso revealed an increase in Kv4.2 (p<0.05).Taken together the study demonstrates that genetic ablation of Kvβ1.1 attenuates Iso induced cardiac injury. This is a first report clearly delineating the pathophysiological role of Kvβ1.1 in the heart.
PPAR‐gamma is required in the myocardium for outflow tract development (699.2)
Authors: Keith Benjamin , Zhou Lun , Xie Linglin
Published: June 2021

One‐third of congenital heart defects involve abnormalities in the development of the outflow tract (OFT). In our lab’s previous study, 100% of PPAR‐gamma heterozygote mouse embryos at embryonic day 14.5 (E14.5) showed OFT defects including ventricular septal defects (VSD), overriding aorta (OA), double outlet right ventricle (DORV), and common arterial trunk (CAT). These observations suggested that PPAR‐gamma plays a vital role in the development of the OFT. To identify the destined cell lineage for PPAR‐gamma in a specific cell lineage, we performed cre‐lox recombination technology based genetic study to knockdown PPAR‐gamma in specific cell lineages including the myocardium (Tnt‐cre), endocardium (Tie2‐cre), and the second heart field (Mef2c‐cre). We found that 33% (2/6) PPAR‐gammaTnt‐cre/+ embryos at E13.5 had OA, but no OFT defects were observed in either PPAR‐gammaTie2‐cre/+ or PPAR‐gammaMef2c‐cre/+. This data revealed that PPAR‐gamma is required in the myocardium, but not endocardium nor second heart field, for the development of the OFT.

Grant Funding Source: Supported by the APS STRIDE summer undergraduate fellowship and the UND faculty seed money
The impact of cycling in a hot environment on cytokine release (705.2)
Authors: Cherif Anissa , Knez Wade , Farooq Abdulaziz
Published: June 2021

The objective of this study was to determine the effect of hyperthermic environment in combination with exercise on the plasma concentration of interleukin‐1ra (IL‐1ra), interleukin‐6 (IL‐6) and C‐reactive protein (CRP), compared with exercise in a neutral condition. Nine recreational volunteers cycled in two different environmental conditions (neutral (24⁰C, 30⁰C) and hot (35⁰C, 70⁰C). Blood samples were collected before, during and after exercise (0, 10, 25, 35, 45, 55 and 65 min) to determine indirect markers of inflammation. ELISA determined the concentration of cytokines and CRP. Overall, the production of IL‐1ra was significantly higher in the hot condition (216.5±71.9 µg/ml) compared to neutral (198.3±49.5 µg/ml) (P=0.003) and increased with time (P=0.009). Similarly there was a significant effect of condition (P<0.001), IL‐6 in hot condition (663.9±270.4 µg/ml) was greater than neutral (499.5±127.8 µg/ml) and CRP in the hot condition (565.2±252.3 ng/ml) was higher than neutral (405.7±221.8 ng/ml) (P<0.001). There were significant effect of time on IL‐1ra (P=0.009) and IL‐6 (P=0.013) but not CRP. Initial analysis suggests that exercising in hyperthermic conditions increase IL‐6, which stimulates the production of the anti‐inflammatory properties of IL‐1ra and CRP.
Aerobic interval training increases circulating number and improves function of monocyte‐derived endothelial progenitor cells in sedentary men (706.5)
Authors: Tsai Hsing‐Hua , Chen Rong‐Rong , Wang Jong‐Shyan
Published: June 2021

This study investigated the effects of aerobic interval (AIT) and moderate continuous training (MCT) on circulating number and function of monocyte‐derived endothelial progenitor cells (EPCs) in sedentary men. Twenty‐four sedentary males were randomized to perform AIT (3‐minute intervals at 40% and 80% VO2 peak, n=12) or MCT (sustained 60% VO2 peak, n=12) for 30 minutes/day, 5 days/week for 6 weeks. Counts of CD34+and kinase domain receptor (KDR)+ monocytes attributed to speciﬁc monocyte subsets [CD14++/CD16‐ (Mon1), CD14++/CD16+ (Mon2), and CD14+/CD16++ (Mon3)] by four‐color flow cytometry. Endothelial function was measured by endothelium‐dependent ﬂow‐mediated vasodilatation (FMD). Late out‐growth EPCs were cultured to evaluate wounding migration and tube formation. The experimental results showed that both AIT and MCT increased circulating level of CD34+/KDR+ Mon2 cells. Moreover, FMD was markedly higher with AIT compared to MCT. However, AIT rather than MCT significantly enhanced in vitro wounding migration and tube formation. Hence we conclude that AIT effectively increases circulating number and improves function of monocyte‐derived EPCs in sedentary men.
Time in culture alters the biophysical responses of rat carotid body cells (709.1)
Authors: Kim Insook , Donnelly Dave , Fite Lasha , Carroll John
Published: June 2021

The carotid body (CB) senses blood O2, CO2, and pH. Fresh CB glomus cells depolarize and increase intracellular Ca2+ ([Ca2+]i) in response to K+, and hypoxia but not to ATP, while non‐glomus cells increase [Ca2+]i in response to extracellular ATP, alone. We determined whether these response characteristics changed in culture. Cells dissociated from postnatal day 15 rat CB were plated and cultured for 7‐8 days. Cultured CB cells were tested for [Ca2+]i responses to 20mM K+, hypoxia, anoxia, TEA(10mM)+4‐AP(5mM), and ATP(100µM) when fresh (0 day culture, C0) and at days in culture C1, C3, C5 and C7. Glomus cells were identified by PNA binding (peanut agglutinin) and absence of response to extracellular ATP. Compared to C0: 1) [Ca2+]i responses to 20mM K+ were not reduced at C1, but were significantly reduced at C3 and C7. 2) [Ca2+]i responses to hypoxia and anoxia in glomus cells were significantly reduced at C1, reduced further at C3 and remained low at C7. 3) Non‐glomus cells (negative PNA labeling, positive ATP response), which initially showed no response to anoxia on C0, developed a [Ca2+]i response to anoxia by C1. 4) [Ca2+]i responses to TEA+4‐AP in glomus cells were not changed during whole 7 day culture period, while the [Ca2+]i response to TEA+4‐AP in non‐glomus cells was significantly increased at C3 and remained so until C7. We conclude that both glomus and non‐glomus cells change their responsiveness in short‐term culture. In particular there is a dramatic decrease in the [Ca2+]i response to hypoxia of glomus cells and development of a [Ca2+]i response in non‐glomus cells to both anoxia and K+ channel blockers.

Grant Funding Source: NIH RO1 HL54621
Implementation of rapid prototyping to produce a 3‐dimensional physical model of the anatomical distribution of functionally identified cat pre‐Bötzinger complex microinjection sites (714.4)
Authors: Geritano Mariah , Lam Sharon , O'Neal Marvin , Cordovez Juan , Solomon Irene
Published: June 2021

Previous work from our laboratory has used histological analyses to confirm functionally identified sites in the cat pre‐Botzinger complex (pre‐BotC). These sites have been represented in a 2‐D fashion based on our histological sections in conjunction with plates from Berman's stereotactic atlas (1968). To gain further insight into the anatomical distribution of these sites, we have used rapid prototyping technology to generate a 3‐D physical model representing the relevant regions/nuclei of the cat brainstem (based on Berman’s atlas) and the histological/stereotaxic coordinates associated with our microinjection sites. To accomplish this, plates from Berman’s atlas were scanned and compiled into a 3‐D configuration slide box, the stereotaxic coordinates of 348 pre‐BötC microinjection sites were entered into an x/y/z matrix based on histologically confirmed coordinates with the origin (0, 0, 0) referenced to the caudal pole of the retrofacial nucleus, midline, and dorsal surface of the medulla, and a 3‐D digital representation was generated and printed. We believe that this approach compliments both the 2‐D representations (previously used) and the digital 3‐D representation, and provides additional insight into the anatomical specificity of our functionally identified and histologically confirmed microinjection sites.

Grant Funding Source: Supported by HL 63175
Ectodomain shedding in the downregulation of cellular ACE‐2 by ER stress in lung cells (716.3)
Authors: Dang Vinh , Uhal Bruce
Published: June 2021

Previous work from this laboratory has demonstrated that angiotensin (ANG) converting enzyme‐2 (ACE‐2) protects against lung fibrosis by limiting the local accumulation of the profibrotic peptide, ANGII by converting ANGII to angiotensin 1‐7 (ANG 1‐7). However, this protective enzyme is down‐regulated in human and experimental lung fibrosis. We currently know of three ways that ACE‐2 is down‐regulated in lung cells: 1) exposure to hyperoxia in fetal lung fibroblasts, 2) treatment with proteasome inhibitors, MG132 or Clasto‐Lactacystin β‐lactone (CLBL) in alveolar epithelial cells (AECs), and 3) expressing the surfactant protein C mutation G100S (which causes lung fibrosis in people). In all of these cases, ACE‐2 was only measured in cell lysates, but not in the cell culture medium to assess “shedding”. We hypothesize that the down‐regulation of cellular ACE‐2 may be mediated by ectodomain shedding by ADAM17/TACE. Western blots of cell lysates revealed a significant decrease in immunoreactive cellular ACE‐2 with CLBL (p= < 0.01) and a potential recovery in ACE‐2 with TAPI‐2, an inhibitor of ADAM17/TACE. Contrary to our hypothesis, a similar effect was also observed in serum‐free cell culture media (p= < 0.05). From this, we can speculate that ADAMS17/TACE may play a partial role in down‐regulating ACE‐2 by degrading it, in addition to other enzymes. With this model, further research can be done to investigate alternative mechanisms that lead to the down‐regulation of cellular ACE‐2 in lung cells.

Grant Funding Source: Supported by APS STRIDE Fellowship and PHS HL ‐ 45136
Neuroanatomy course: a nightmare for Nigerian medical students (719.16)
Authors: Musa Muhammad , Danfulani Muhammad , Bello Sanusi
Published: June 2021

Appreciating basic neuro‐anatomy by medical students plays a vital role in their understanding of the pool of neurological diseases and illnesses found in Nigeria and sub‐Saharan Africa. This study was conducted to ascertain the perception of Nigerian medical students about neuro‐anatomy in comparison with other aspects of human anatomy course. The study participants are third year students from 2 medical schools in Northwestern Nigeria. Anonymous and voluntary questionnaire was distributed to the randomly selected participants. To date, out of the 120 questionnaires distributed, a total of 116 have been completed and returned. Information obtained has been entered into Microsoft Excel Worksheet after which relevant descriptive and inferential statistics will be applied. Preliminary findings from the completed and returned questionnaires, revealed that majority of the medical students perceived neuro‐anatomy course as the most difficult aspect of human anatomy. So far, 60% of the respondents consider poor teaching methodology employed by the neuro‐anatomy tutors as the main reason for their negative perception and suggested the use of models as well as teaching aids to enhance learning. This study would probably give a hint as to why most medical graduates never considered picking up careers in neurology or neuroscience related fields, thus making it difficult to address the health challenges noticed in these fields of endeavors.
Incorporation of autonomic pharmacology into high fidelity simulated cardiopulmonary critical care scenarios improves cardiopulmonary pharmacology assessment performance (719.3)
Authors: Gorman A. Laurel , Gros Bernard , Johnson Teresa , Cendan Juan
Published: June 2021

Effective teaching of autonomic pharmacology is essential for understanding the pathology and treatment of cardio‐pulmonary disease. To improve comprehension, we incorporated sympathomimetic treatments into high fidelity medical simulations (HFMS) involving scenarios of shock and respiratory insufficiency. Groups of five students treated their SimMan manikin, with faculty scaffolding the experience. Multi‐disciplinary debriefings performed after HFMS focused on integrated clinical and pharmacology teaching. Students performed poorly on pre‐HFMS quizzes after didactic sessions covering autonomic pharmacology (Mean: 54%) but improved significantly after the HFMS (Mean: 84%). Students exposed to HFMS performed better on summative autonomic pharmacology questions (Mean: 89%) than a cohort population of similar M2 students not taught autonomic pharmacology in HFMS (Mean: 74%). While the cohort with the lower score received 4 hours of topic didactics, the higher scoring HFMS cohort received 1.5 hours of topic didactics. Therefore, teaching autonomic pharmacology using HFMS improved acquisition of autonomic pharmacology despite less didactic hours being allotted. Assessment outcomes support student perceptions, with 95% of students agreeing that HFMS improved pharmacology understanding. Thus, HFMS is an effective and efficient pedagogy for teaching preclinical autonomic pharmacology.
Perceptions of professional student readiness for physiology courses: can online review modules help? (720.3)
Authors: Miller Cynthia , Metz Michael
Published: June 2021

There can be a disconnect between the level of content covered in undergraduate coursework and the expectations of professional‐level faculty of their incoming students. Some basic science faculty members may assume that students have a good knowledge base in the material and neglect to appropriately review, while others may spend too much class time reviewing basic material. It was hypothesized that the replacement of introductory didactic physiology lectures with interactive online modules in a first‐year D.M.D. course could improve student preparedness prior to lectures. These modules would also allow faculty members to analyze incoming student abilities and save valuable face‐to‐face class time for alternative teaching strategies. Results indicated that the performance levels of incoming students was poor (57% average on a pre‐test), and students often under‐predicted their abilities (by 13% on average). Faculty expectations varied greatly between the different content areas, and did not appear to correlate with the actual student performance. Three review modules were created which produced a statistically significant increase in post‐test scores (46% increase, p<.0001, n=114‐115). The positive results of this study suggest a need to incorporate online review units in dental physiology courses and revise introductory material tailored to students’ strengths and needs.
Give histology some TLC: Team Laboratory Chiefs (721.15)
Authors: Reidenberg Joy , Shaw Phyllis
Published: June 2021

The Histology Team Laboratory Chief (TLC) concept was developed to foster team‐based learning, institute peer‐teaching and peer‐grading, and facilitate using the “Virtual Histology” digital microscope slide program (developed by co‐author P.A.S.). Each student led six labs to a six‐member team as one of a pair of TLCs. Presentation format was flexible, allowing creativity and incorporation of additional resources. After two “ramp up” sessions in which students worked independently to learn “Virtual Histology,” TLC partners presented assigned slides, focusing on important distinguishing structural features and functions of cells/tissues/organs. TLCs were required to teach the relevant content of the laboratory, ask four questions to facilitate discussion, and answer questions to clarify issues and assure appropriate understanding of structure and function. Faculty rotated among teams, answering questions and correcting any errors. In 2013, anonymous on‐line scoring was introduced to prevent peer‐pressure grade inflation. The average TLC score was 97.2%, indicating high satisfaction with presentations. Students agreed or strongly agreed that the TLC program increased understanding of course material (70%), and that teaching peers increased their teaching skills (67%). The TLC program contributed to a dramatic 47% increase from 2009 in satisfaction that lab sessions improved comprehension.
Sonoanatomy in a team‐based combined medical and physician assistant curriculum (722.2)
Authors: Hartley Rebecca , McGuire Paul , Kalishman Summers , Sklar David , Rosett Randy , Vagh Firoz
Published: June 2021

A curriculum in sonoanatomy was piloted in the Human Structure, Function and Development (HSFD) block at the University of New Mexico. This 10‐week block combines anatomy, histology and embryology using team‐based learning. It is the first block in the 1.5 year integrated Phase I Medical and Physician Assistant (PA) curriculum. The purpose was to expose the 111 Medical and 17 PA students to ultrasound, which is becoming essential in the clinic, by providing hands on 3D imaging to reinforce and apply anatomy knowledge. Three 1‐hour sessions were led once per week by two clinicians, with 20‐22 students per instructor per session. Sessions correlated with the weekly laboratory objectives and began with a short introduction and an ultrasound demonstration on a standardized patient. Each student then practiced ultrasonography on the standardized patient. Ultrasound images were included on weekly individual and group readiness assessment tests, clinical application cases, and three section exams. Performance on ultrasound questions varied on summative (better) and formative (worse) questions and also differed dependent on anatomical region. Student evaluations indicated that the ultrasound sessions supported better understanding of the content each week (3.23/5 points on a Likert scale of agreement). Students especially appreciated clinician interaction (4.22/5 points). Specific student comments will be used to improve and better integrate sonoanatomy with HSFD.
The integration of osteology, radiology, and surface anatomy into a Clinical Skills 101 course for first‐year medical students (723.1)
Authors: Giffin Bruce , Lowrie D.J. , Pettigrew David , Gunderson Anne
Published: June 2021

Clinical Skills 101 is a new course with the goal of imparting patient‐ centered skills fundamental to promoting health and treating disease. Teaching the physical examination requires knowledge of surface anatomy, and understanding the variation of normal visible and palpable anatomy enables the physician to determine when pathology may be responsible for abnormalities in surface anatomy or underlying structures. Since the skeletal system elements are responsible for many of the palpable bony contours seen on the surface of the body, preliminary knowledge of the skeletal system is essential for learning surface anatomy. Students are taught the basics of the physical examination using standardized patients in conjunction with surface anatomy and osteology laboratory exercises. Since medical imaging has come to occupy a central role in the delivery of patient care, imaging exercises reinforce the osteology and demonstrate important imaging features of common or important pathologies. Weekly assessment of the students' progress in the acquisition of surface anatomy and imaging skills consists of surface anatomy testing using fellow students as “patients”, osteological practical exams, and identification of normal and abnormal imaging. Evaluation of this integrated approach to the acquisition of clinical skills is highly rated by students and clinical preceptors.
Development and evaluation of a computer‐aided learning resource in anatomy (725.6)
Authors: Russell Rae , Lee Lisa
Published: June 2021

Teaching professional health science students complex anatomy effectively requires the use of visual aids such as dissection, diagrams, and atlases. The challenges of the structural and functional complexities of the central nervous system, coupled with the reduced basic science curricular hours, necessitated utilizing computer‐aided instruction. The objective of the study is to promote mastery of the spinal cord anatomy by developing an efficient online ancillary resource for studying gross anatomy. A 23‐minute online learning module on spinal cord anatomy was constructed, presenting images of cadaveric dissections and diagrams. The effectiveness of the module and its perceived educational value were assessed on a naïve population of dental students before the gross anatomy course. The results reveal a significant amount of learning occurred based on comparative pre‐quiz and post‐quiz scores. The survey yielded positive perception of educational value however a few areas of improvements were also identified. The user‐tested and accordingly modified learning modules are expected to make anatomy education more accessible and effective in the currently limited student‐instructor contact hours.
Influence neurotransplantation on the enzymatic activity of cells of the cerebral cortex of rats resuscitated different gender and age (729.1)
Authors: Tazhibaeva D.C. , Kabdualieva N.B. , Aitbayeva Zh. , Mirzakhanov N. , Mirzakhanova M.
Published: June 2021

The objectives of the research included:Study the comparative aspects of the dynamics of enzyme activity of the cerebral cortex reanimated rats of different age and sex using fetal nerve cells;Experiments have been performed on 43 rats of both sex, who turned 1,5 months and 12 months by the research beginning. A 4 minute clinical death from acute mechanical asphyxia was simulated. At the time of cardiac activity restoration we performed intraperitoneal injection of fetal cells of the nervous tissue in a dose of 1mm per 25g of the animal. Repeated injection of the cells was carried out on 14th day of the postresuscitational period. The rate of rats’ survival was determined in the postresuscitational period. For the purposes of research of the kinetics of enzymatic reactions in the homogenate of the cerebral cortex the content of the following enzymes was determined (Tell L.Z.): lactate dehydrogenase (LDH), creatine kinase (CK), alkaline phosphatase, cholinesterase (ChE). As a result of the single neurotransplantation the males has improvement in an enzymatic activity of the cerebral cortex, whereas enzymatic activity of the females get worse. Repeated neurotransplantation is accompanied by reduction in activity Che in young reanimated males, young females have declined enzymatic activity of LDH and increase ‐ CK. In adult females, along with decreased activity of creatine kinase increases the enzymatic activity cholinesterase.
Expression of functional light‐activated ion channels in embryonic chick muscle (731.1)
Authors: Whitaker Jessica , Sharp Andrew
Published: June 2021

Optogenetic tools allow for fine control of targeted populations of neurons. Muscle cells are an electrically excitable cell type that could be targeted for regulation by optogenes. Our lab studies the effects of altered embryonic movement and sensory feedback on the neural development of chick embryos. Previously, we have used electroporation of a modified transposon‐based expression system into the neural tube of embryos to introduce excitatory and inhibitory light‐activated ion channels into neural tissue (Sharp and Fromherz, 2011). Similar to neurons, muscle cells can be induced to contract in response to changes in membrane potential. Therefore, we have transformed muscle precursor cells in the somites of embryonic day (E) 3 chick embryos with the goal of being able to control the activity of muscles in ovo. We have observed expression of ChR2 in muscle of intact embryos and sectioned tissue via a fluorescent reporter molecule as late in development as E16. Sectioned muscle tissue reveals patterns of fluorescence consistent with membrane expression of ChR2. Further, we have demonstrated the ability to cause hip flexion when ChR2 is expressed in the sartorius muscle. Upon exposure to pulses of blue light, the hip flexed with and average onset latency of 0.137 s. Hip flexions stopped after five or six consecutive pulses. This approach will allow for the modulation of muscle activity during embryogenesis.

Grant Funding Source: Supported in part by a Research‐Enriched Academic Challenge Award to J.W. from SIUC.
Myofiber damage precedes macrophage infiltration after contraction‐induced injury in dysferlin‐null muscle (731.3)
Authors: Roche Joseph , Tulapurkar Mohan
Published: June 2021

Mutations in the DYSF gene that encodes the protein, dysferlin, lead to late‐onset, human muscular dystrophies known as dysferlinopathies. The A/J mouse spontaneously develops a mild dysferlinopathy after 6 months of age and at younger ages models the subclinical phase of the human disease. We subjected 3‐4 month old A/J mice to in vivo large‐strain lengthening injury (LSI) from lengthening contractions and studied the role of inflammation in the onset of muscle damage and recovery thereafter. We show that post‐LSI, myofiber damage in the tibialis anterior muscle occurs prior to inflammatory cell infiltration. Indeed, most edema and inflammation, monitored by T2‐weighted MRI and by immunofluorescence labeling of neutrophils and macrophages, respectively, develops 24‐72 hr after LSI, well after the appearance of damaged myofibers. The cytokines IL‐1A, MCP‐1, and MCSF are elevated several‐fold at 72 hr after injury, consistent with extensive macrophage infiltration. Control A/WySnJ mice show much less myofiber damage, much less inflammation and much lower cytokine levels after LSI than A/J. Depletion of monocytes and macrophages by systemic clodronate administration, fails to suppress myofiber damage or to accelerate functional recovery in A/J mice. Our studies suggest that, although macrophage infiltration is prominent in A/J muscle following LSI, it is the consequence and not the primary cause of progressive myofiber damage.

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Grant Funding Source: Supported by a grant to JAR from the Jain Foundation Inc.