bluuebunny/crossref_metadata_2025 huggingface dataset:
Epigallocatechin‐3‐Gallate inhibits the effects of TGF‐Beta and IL‐1B on ENaC expression in human alveolar epithelial cells.
Authors: Phillips Oluwakemi , Cox Ruan , Fukumoto Jutaro , Fukumoto Itsuko , Parthasarathy Prasanna Tamarapu , Delic Vedad , Luong Tran , Lockey Richard , Kolliputi Narasaiah
Published: June 2021

Acute Lung Injury (ALI) is major clinical problem in the United States, affecting over 200,000 Americans a year. Increased mortality in ALI is attributed to pulmonary edema and decreased net fluid transport. Previous clinical studies from ALI patients have correlated the decrease net fluid transport with decreased expression of epithelium sodium channels (ENaC). Therefore drugs that enhance ENaC expression offer considerable therapeutic promise in ALI. Epigallocatechin‐3‐Gallate (EGCG), a green tea extract, has been shown to have potent anti‐inflammatory effects. However, its effect on ENaC expression in response to proinflammatory cytokines in ALI has not been shown. In this study, we investigated the hypothesis that EGCG rescues human alveolar epithelial cell (A549) ENaC expression following proinflammatory cytokine (IL‐1β and TGFβ) treatment. To test this, A549 cells were treated with TGF‐β and IL‐1β in the presence or absence of EGCG. Cell lysates as well as supernatants were collected for protein analysis. EGCG treatments resulted in increased expression of the alpha and beta subunits of ENaC in response to both TGF‐ β and IL‐ β. Further, EGCG treatments resulted in decreased cytokine secretion mediated through decreased NF‐kappaB and p‐38 MAP kinase activity in comparison to control. Our data suggests that EGCG up regulates ENaC, a major sodium channel involved in resolution of pulmonary edema in ALI.
Microtubule stability is affected by oxidative stress and contributes to lung injury and barrier dysfunction
Authors: Birukova Anna A , Tian Yufeng , Sarich Nicolene , Meliton Angelo
Published: June 2021

Pathologic activation of redox signaling by inflammatory agents causes endothelial cell (EC) activation and barrier dysfunction. Microtubules (MT) play an important role in the regulation of vascular endothelial permeability, but their impact in modulation of inflammation and EC barrier is not well understood. This study examined effects of LPS‐induced oxidative stress on MT dynamics and involvement of MT in the LPS‐induced mechanisms of Rho GTPase activation, EC permeability and lung injury. LPS treatment of pulmonary vascular EC induced elevation of reactive oxygen species (ROS) and oxidative stress leading to cytoskeletal remodeling, EC hyper‐permeability, increased neutrophil adhesion and activation of inflammatory signaling. LPS also triggered ROS‐dependent disassembly of the microtubule network leading to activation of MT‐dependent signaling. Stabilization of MT or inhibition of MT‐associated Rho‐specific GEF activity suppressed LPS‐induced EC barrier dysfunction in vitro and attenuated vascular leak and lung inflammation in vivo. These studies demonstrate for the first time the mechanism of ROS‐induced Rho activation via destabilization of MT and GEF‐H1‐dependent activation of Rho signaling leading to pulmonary EC barrier dysfunction and exacerbation of LPS‐induced inflammation.

Funding: NHLBI HL089257, HL107920
Hyperoxia induced lung injury is associated with alterations in circadian clock genes in mice
Authors: Lagishetty Venu , Parthasarathy Prasanna Tamarapu , Huynh Bao , Fukumoto Jutaro , Cox Ruan , Lockey Richard , Kolliputi Narasaiah
Published: June 2021

Prolonged exposure to hyperoxia leads to inflammation and hyperoxia‐induced acute lung injury(HALI).The master regulators of circadian rhythm, clock genes, have been implicated in inflammation and chronic lung diseases. However the role of clock genes in HALI has not been studied. This study was designed to determine whether HALI is mediated by altered expression of clock genes.To test our hypothesis, mice were exposed to hyperoxia or room air(control)for 72h. Lung clock gene mRNA and protein levels were analyzed by qPCR and western blot analysis respectively. DNA methylation of Per1 gene promoter was measured by methyl specific PCR. mRNA and protein levels of clock genes(CLOCK, Bmal1, Per1&2, Cry1&2) and their target genes(Rev‐erba, PPARg) were significantly elevated in mice exposed to hyperoxia compared to controls. Alterations in clock genes were associated with increased inflammatory markers (Nfkb, IL1b, TNFa and NALP3) in hyperoxic mice compared to controls. Further, promoter region of Per1 gene in hyperoxic mice was hypomethylated compared to control. Together, our data demonstrates that increased inflammation in HALI may be partly due to hypomethylation of Per1 and increased clock gene expression.
Angiotensin Receptor Blockade Reduces Oxidative Stress while Improving Redox Signaling and Mitochondrial Function in the Heart of Diet‐induced Obese Insulin Resistant Rats
Authors: Vazquez‐Medina Jose Pablo , Popovich Irina , Thorwald Max A , Rodriguez Ruben , Viscarra Jose A , Lam Lisa , Peti‐Peterd Janos , Nakano Daisuke , Nishiyama Akira , Ortiz Rudy M
Published: June 2021

The contributions of angiotensin receptor type 1 (AT1) activation to NADPH oxidase (Nox)‐derived oxidative stress during hypertension and diabetes are well established. The role of AT1 in modulating redox signaling, mitochondrial function and oxidative stress in the heart during metabolic syndrome, however, remains elusive. To test the hypothesis that AT1 activation increases oxidative stress while impairing mitochondrial function and redox signaling in the heart during metabolic syndrome, diet‐induced obese, insulin resistant (OLETF) rats were treated with the AT1 blocker (ARB) Olmesartan for 6 weeks. Cardiac Nox2 expression increased 40% in OLETF compared to age‐matched lean (LETO) rats while expression of the H2O2‐producing Nox4 increased 40%. ARB treatment prevented the increase in Nox2 without altering Nox4. ARB treatment also normalized increased protein and lipid oxidation and increased the redox‐sensitive transcription factor Nrf2 30%, and antioxidant enzymes by 50–70%. Mitochondrial aconitase activity increased 60% in OLETF compared to LETO whereas succinate decreased 35%; ARB treatment normalized both. These data demonstrate that AT1 activation contributes to increased Nox2‐derived oxidative stress and to impaired redox signaling and mitochondrial activity, and suggests that, besides its anti‐hypertensive effects, ARB treatment may improve myocardial function during metabolic syndrome.
MCPIP‐1 deficiency induces hepatic inflammation and impairs insulin signaling in mice
Authors: CHANG YINGZI , Holdaway Brett Benson , Moody Joshua Dan , Fu Mingui , Kolattukudy Pappachan
Published: June 2021

Emerging evidence suggests that elevated inflammatory/immune events are associated with pathogenesis of diabetes. MCPIP1 (monocyte chemotactic protein [MCP]–induced protein 1; also known as ZC3H12A), an immune response modifier expressed in mice macrophage in response to LPS and MCP‐1 stimulation, plays a negative role in the process of inflammation. Our current study was designed to test if MCPIP‐1 deficiency induced metabolic dysfunction by affecting the insulin signaling transduction pathway. Wild‐type C57/BL6 mice and MCPIP‐1 knockout mice with the C57/BL6 background were used. We found that MCPIP‐1‐deficient mice showed severe hepatic inflammation and remodeling, lower fasting plasma insulin levels (p<0.05), delayed response to the glucose challenge (p<0.05), and reduced insulin tolerance (p<0.05). Our results also revealed that insulin‐induced phosphorylation of insulin receptor (IR), insulin receptor substrate 1(IRS‐1), and AKT were substantially decreased in the liver from MCPIP1‐deficient mice. In conclusion, MCPIP‐1 deficiency impairs insulin production, insulin signaling transduction, and the ability of enduring metabolic challenges by inducing hepatic inflammation.

Funded by Warner's grant, A.T. Still University of Health Sciences.
Diurnal Variation in Mouse Urinary Bladder Smooth Muscle (UBSM) Contractility
Authors: Herrera Gerald M. , White Rachel , Meredith Andrea L.
Published: June 2021

Physiological output varies by time of day, coordinated by the central circadian clock in the hypothalamus. Recently, a circadian rhythm in micturition was demonstrated in rodents, correlated with a day‐night change in the storage and voiding of the bladder. The goal of this study was to determine whether isolated UBSM exhibited an intrinsic difference in contractility between day and night. Mouse UBSM strips were harvested at 4 time points over the circadian cycle, zeitgeber time (ZT) 2, 8, 14, and 20, and phasic and nerve‐evoked (EFS) contractions were investigated using isometric tension recordings. We hypothesized that UBSM harvested from mice during the active period when micturition peaks (ZT8–14) would demonstrate stronger contractile activity than strips harvested during the rest period (ZT20–2), when the bladder relaxes to store urine. Compared to ZT20, at ZT8 UBSM strips exhibited larger KCl‐induced (10 ± 0.4 versus 7 ± 0.4 mN), phasic (0.08 ± 0.02 versus 0.03 ± 0.01 mN), and EFS‐induced contractions (28 ± 3 versus 22 ± 2 mN, n = 6 mice per condition). Increased contractile amplitudes at ZT8 were correlated with lower expression of the BK K+ channel, a regulator of UBSM excitability. These results identify a diurnal difference in UBSM contractility in the absence of central control, and suggest that daily regulation of bladder excitability occurs locally within smooth muscle tissue. Funded by NIDDK R21089337.
CaV3.2 Channels and the Induction of Negative Feedback in Cerebral Arterial Smooth Muscle
Authors: El‐Rahman Rasha Abd , Harraz Osama , Bigdely‐Shamloo Kamran , Mufti Rania , Gonzales Albert , Earley Scott , Vigmond Edward , Wilson Sean , Welsh Donald
Published: June 2021

CaV1.2 (L‐type) along CaV3.1/CaV3.2 (T‐type) are the principal subtypes of voltage‐gated Ca2+ channels (VGCC) expressed in cerebral arterial smooth muscle. While studies have long discerned the functional role of CaV1.2, the physiological significance of CaV3.x expression is uncertain. Recent immunohistochemical analysis noted that CaV3.2 localizes in close proximity to ryanodine receptors (RyR) on the sarcoplasmic reticulum. From these observations, we hypothesized that CaV3.2 triggers Ca2+ ‐induced Ca2+ release (RyR), activating large‐conductance Ca2+‐activated K+ channels (BKCa) to attenuate arterial constriction. Structural analysis involving immuno‐labeling approaches, electron microscopy, and 3D‐tomomography revealed a microdomain structure in cerebral arteries comprised of CaV3.2 and RyR. Using mathematical techniques, a microdomain model was subsequently developed and it revealed that Cav3.2 was capable of activating RyR and induce repetitive CICR‐like events. In keeping with these theoretical observations, perforated patch clamp electrophysiology revealed that Ni2+ (50 μM, Cav3.2 inhibitor) attenuated the frequency and amplitude of BKCa‐mediated spontaneous transient outward currents (STOCs). Pressurized cerebral arteries were also shown to depolarized and constricted to micromolar Ni2+. The magnitude of these functional responses was comparable to paxilline, a BKCa channel inhibitor. In summary, findings indicate for the first time that CaV3.2 channels are capable of driving a CICR‐like process that moderates arterial constriction through a feedback response involving BKCa channels.
Can the study of individuals with autonomically complete spinal cord injuries help clarify the role of sympathetic nerves in cerebrovascular reactivity?
Authors: Inskip Jessica , Ravensbergen Rianne H.J.C. , O'Connor Shawn M. , Claydon Victoria E.
Published: June 2021

Cerebral blood vessels are richly innervated by sympathetic nerves; however, their role in the control of the cerebral vasculature remains controversial. Individuals with spinal cord injury (SCI) provide a unique opportunity to study cerebrovascular control in the absence of sympathetic control. Here we studied cerebrovascular reactivity to CO2 in individuals with cardiovascular autonomically complete and incomplete SCI, and neurologically intact controls. Cardiovascular autonomic completeness was assessed on the basis of supine plasma noradrenaline levels, blood pressure variability and lesion level. Supine beat‐to‐beat blood pressure and middle cerebral artery velocity (MCAv) were continuously recorded, as were breath‐by‐breath partial pressures of end‐tidal O2 and CO2 (PETCO2), during manipulation of PETCO2. Low levels of PETCO2 were obtained by voluntary hyperventilation, and high levels by hypoventilation with increased inspired CO2, during normoxia (controlled on a breath‐by‐breath basis using dynamic end‐tidal forcing). MCAv reactivity was significantly different between groups (control 11.2±2.4; autonomically incomplete SCI 5.8±0.6; autonomically complete SCI 6.2±0.7 %/mmHg, p=0.025, mean±sem). These preliminary data suggest that there is impaired cerebrovascular reactivity in individuals with SCI that is independent of injury to cardiovascular autonomic pathways.

This research is supported by the Heart and Stroke Foundation of B.C. and Yukon (V.E.C).
ATP‐induced sympathoexcitation involves purinergic and glutamatergic mechanisms in the RVLM‐projecting PVN neurons
Authors: Neto Hildebrando Candido Ferreira , Antunes Vagner Roberto
Published: June 2021

We have shown that ATP applied into the PVN induces a dose‐dependent increase in the lumbar sympathetic nerve activity (LSNA), such effect attenuated by the antagonism of P2 and/or ionotropic glutamate receptors within the PVN. In this study we aimed to evaluate whether the sympathoexcitatory responses elicited by purinergic activation within the PVN would involve the release of ATP and/or glutamate at the RVLM level. Recordings of LSNA were monitored decorticated and arterially perfused rat preparation. Three‐barrelled glass pipettes were filled with ATP (2.5 nmol), PPADS (0.5 nmol) and kynurenic acid (KYN, 10 nmol) for injection into the PVN and/or into the RVLM. Unilateral microinjection of ATP into the PVN induced an increase in the LSNA (control 100%), which was significantly attenuated 5 min after ipsilateral injection of KYN into the RVLM (59±7%, n=6, p<0.05), with recovery to control level 30 min later (100±15%). ATP‐induced sympathoexcitation in the PVN was also significantly attenuated 10 min after ipsilateral injection of PPADS within the RVLM (65±9%, n=9, p<0.05) with recovery 30 min later (98±14%). Our data suggests that P2 and glutamatergic receptors in the RLVM participate of the ATP‐induced sympathoexcitation into the PVN and this purino‐glutamatergic neurotransmission in the hypothalamic‐brainstem circuitry in a physiological context awaits further investigation. Sponsored by CAPES, CNPq, FAPESP.
DELETION OF PROTEIN TYROSINE PHOSPHATASE 1B PROTECTS AGAINST ENDOPLASMIC RETICULUM STRESS‐INDUCED ENDOTHELIAL DYSFUNCTION
Authors: Norman Joseph Blake , Herren David , Butler Benjamin , Tremblay Michel L , Belin de Chantemele Eric J.
Published: June 2021

Protein Tyrosine Phosphatase 1B (PTP1B) modulates endoplasmic reticulum stress (ERS). However, whether PTP1B regulates ERS‐nduced endothelial dysfunction is unknown. To test the hypothesis that PTP1B deletion protects against ERS‐mediated endothelial dysfunction, ERS was induced with either tunicamycin (TUNICA) in aortic rings from PTP1B null (KO) and control mice (WT) or by treating WT and KO mice with STZ for 4 weeks. Assessment of the vascular function demonstrated that TUNICA reduced endothelial function by 30% in WT mice while endothelial function remained intact in KO mice. ACh relaxation was completely abolished by LNAME but neither tempol nor indomethacin improved ERS‐mediated endothelial dysfunction, suggesting that neither ROS nor COX derivatives are involved in the dysfunction. TUNICA blunted Ca2+‐mediated relaxation in WT mice only, suggesting that PTP1B deletion preserves eNOS sensitivity to Ca2+. Four weeks of STZ treatment significantly reduced endothelial function in WT mice. Acute treatment of aortic rings with the ERS inhibitor TUDCA restored endothelial function and confirmed that type I diabetes induces endothelial dysfunction via ERS. Despite a higher glycemia compared to WT mice, PTP1B KO mice did not develop an endothetlial dysfunction in response to STZ and TUDCA did not affect vascular relaxation. Consequently, PTP1B appears to be a key target to prevent endothelial function.
Spinal PKC‐θ activity is required for phrenic long‐term facilitation after acute intermittent hypoxia
Authors: Devinney Michael J , Fields Daryl P , Mitchell Gordon S
Published: June 2021

Acute intermittent hypoxia (AIH) elicits spinal, serotonin‐dependent long‐term facilitation of phrenic motor output (pLTF). Because the necessary serotonin type 2 receptors are Gq‐coupled metabotropic receptors, we hypothesized that spinal protein kinase C (PKC) activation is necessary for pLTF. PKC inhibitors were delivered intrathecally at C4 in anesthetized, paralyzed, and ventilated rats 5–15 min before AIH. Vehicle‐treated rats exposed to AIH exhibited pLTF (48±3%, P<0.05, n=15) vs. time controls (10±2%, n=21). pLTF was unaffected by the broad spectrum PKC inhibitors NPC‐15437 (NPC; 54±5%, P>;0.05, n=6) or Go 6983 (52±4%, P>;0.05, n=7), but was attenuated by bisindolymalemide I (BIS; 22±6%, P<0.05, n=5). Because BIS is documented to inhibit PKC‐θ, while NPC and Go 6983 are not, we tested a refined hypothesis that pLTF requires PKC‐θ activity. Pretreatment with either sotrastaurin, a broad‐spectrum inhibitor with potent PKC‐θ effects (17±6%, P<0.05, n=4), or a myristolated pseudosubstrate peptide targeting PKC‐θ (10±6%, P<0.05, n=5) attenuated pLTF. Preliminary studies identified PKC‐θ in ventral cervical spinal homogenates via immunoblot; immunofluorescence indicates expression within identified phrenic motor neurons. Thus, PKC‐θ activity is uniquely required for pLTF, a novel insight into mechanisms of AIH‐induced spinal, respiratory plasticity.

Supported by NIH HL080209, T32 007654, T32GM007507
O‐GlcNAcylation Modulates Hippocampal Plasticity and Alters Hippocampal Dependent Learning and Memory
Authors: Wang Kai , Taylor Erica , Nelson Amy , Bredemann Teruko , Clinton Sarah , Marchase Richard , Chatham John , McMahon Lori
Published: June 2021

O‐GlcNAcylation is a dynamic posttranslational modification involving O‐linked addition of β‐N‐acetylglucosamine (GlcNAc) to serine/threonine residues on nucleocytoplasmic proteins. Hippocampal neurons express high levels of OGT and OGA, enzymes that add and remove O‐GlcNAc moieties. We investigated whether increasing O‐GlcNAcylation, with glucosamine or the OGA inhibitor Thiamet‐G, modulates synaptic transmission at CA3‐CA1 synapses in hippocampus, a brain region required for learning and memory. We find that increased O‐GlcNAcylation induces a novel long‐term depression (glucosamine: 85% ± 2.6% and Thiamet‐G: 81% ± 4.1% of baseline) and causes decrementing long‐term potentiation (control: 137 ± 4%, Thiamet‐G: 122 ± 6% of baseline). We also find that AMPA receptor GluA2, but not GluA1, subunits are O‐GlcNAcylated. We further demonstrate that increasing O‐GlcNAcylation in vivo interferes with novel object recognition while contextual fear conditioning is intact. These findings support the notion that an acute change in O‐GlcNAcylation of synaptic proteins is a novel fundamental mechanism capable of modulating synaptic efficacy and memory processing.
Muscle RING finger‐1 (MuRF1) inhibits thyroid hormonedependent cardiomyocyte growth in vitro and in vivo
Authors: Wadosky Kristine M. , Hite Rebecca L. , Portman Michael A. , Gerdes A. Martin , Willis Monte S.
Published: June 2021

Recent studies in our laboratory have established that MuRF1 regulates the nuclear export of critical nuclear receptors. To define MuRF1's regulation of other relevant nuclear receptors, we investigated its role in thyroid hormone (TH)‐dependent cardiac hypertrophy. The cardiomyocyte‐derived HL‐1 cell line was treated with 1μM triiodothyronine (T3) after MuRF1 expression was increased or transiently knocked down using adenovirus constructs. HL‐1 cells with MuRF1 knockdown exhibited a 48.0% increase in area, compared to control cells having a 17.5% increase (p<0.05). Conversely, increased MuRF1 expression completely inhibited the T3 response in HL‐1 cells (p<0.05 vs. control cells). To determine the applicability of these findings in vivo, MuRF1 transgenic mice were challenged with 1 mg/kg T3 daily for 14 days. Echocardiographic analysis demonstrated that T3 enhanced the anterior and posterior left‐ventricular wall hypertrophy (+37.3% and +39.1%) in wild type mice, while sibling MuRF1 Tg+ mice exhibited a significantly less robust response (+14.6% and +15.8%, respectively; p<0.05). Heart weight to body weight analysis determined that wild type hearts increased 92.1%, while MuRF1Tg+ hearts only increased 56.4% (p<0.05). Understanding MuRF1's regulation of TH‐dependent cardiac hypertrophy may help develop strategies to enhance the beneficial effects of TH repletion therapy in heart failure.
Growth dynamics of the medullary cavity in the rostrum of juvenile paddlefish (Polyodon spathula)
Authors: Mooney Rachel J. , Robertson John
Published: June 2021

The prominent rostrum of the paddlefish has electroreceptive and mechanosensory functions. Not present in newly hatched paddlefish, the rostrum quickly grows to about one‐third the total body length of juvenile fish. Internally, the rostrum has a hyaline cartilage core that contains an adipose‐filled medullary cavity – thus resembling the long bones of vertebrates that have cavities filled with yellow marrow. To better understand formation, growth and function of the rostrum and medullary cavity, we compared cavity volume to rostrum length and total body length in six groups of size‐matched juvenile paddlefish. Medullary cavity volume was modeled as a simple cone. Each rostrum was mid‐sagittally sectioned and imaged, and cavity length and width measurements were obtained using ImageJ. Results show significant differences in ratios of cavity volume:rostrum length and cavity volume:body length between the developmental groups. In early development the medullary cavity increases in size rapidly, later the rate of cavity volume increase slows relative to body and rostrum growth. Elucidating the pattern of growth of the medullary cavity contributes to understanding of the functional anatomy of the rostrum and the biology of paddlefish.
Beneficial effect of melatonin and forced exercise on degeneration of lower motor neuron after focal cerebral ischemia in rats
Authors: Lee Seunghoon , Hong Yunkyung , Hong Yonggeun
Published: June 2021

The stroke lesion primarily affects the territory of the upper motor neuron, but these clinical signs take shape by means of the skeletal muscle controlled by the lower motor neuron. Therefore, we elucidated that melatonin and exercise is a possible intervention on the rescue of lower motor neuron degeneration after focal cerebral ischemia. Following the MCAo, the 8‐week‐old Sprague‐Dawley (SD) rats were treated twice daily with MT (10 mg/kg) at 7:00 and 19:00 hr and control group were treated with physiological saline (vehicle) instead of MT. Also, the rats were trained twice daily with Ex (20 m/min) at 17:00 and 22:00 hr. MT+Ex is significantly inhibited the attenuation of lower motor neuron during 7 days in both cervical and lumbar spinal cord (p<0.01). Analysis of gene expression in gastroncnemius, MT+Ex is not only downregulated the atrophy‐related genes, including MuRF1 and MAFbx but also upregulated the hypertrophic signal upstream molecule, such as IGF‐1 via overexpression of IGF‐1R (p<0.05, p<0.01, respectively). Moreover, there is significantly inhibited the reduction of muscle fiber diameter following MT and MT+Ex (p<0.01). In here, we found that melatonin and forced exercise could be suppressed the lower motor neuron degeneration as well as stroke‐induced muscle atrophy on early stage after MCAo. Accordingly, we suggest that melatonin has a possible rehabilitative agent as well as the synergistic effect with therapeutic exercise.
Regulation of Tfam expression at the onset of muscle disuse
Authors: Tryon Liam Dennis , Pastore Steven F. , Hood David A.
Published: June 2021

Chronic muscle disuse, such as that produced by long‐term denervation, results in a loss of muscle mass in addition to reductions in mitochondrial content and function. However, acute alterations in the expression of factors regulating mitochondrial content, namely mitochondrial transcription factor A (Tfam), have yet to be elucidated following denervation. Thus, to investigate changes in Tfam expression, we employed the in vivo transfection of a Tfam promoter‐luciferase reporter construct into rat tibialis anterior muscle denervated for 0, 8, 16, 24, 72 or 120 hours. Tfam transcription was reduced by 40–70% between 8 and 24 hours of denervation. This was reflected in a 30% reduction in Tfam mRNA content by 24 hours of denervation, which occurred without any notable changes in Tfam mRNA stability. These changes in Tfam transcription and mRNA preceded alterations in mitochondrial content and muscle mass, first noted by 72 and 120 hours, respectively. Our data suggest that denervation results in early changes in gene expression regulating mitochondrial content, which precede subsequent decrements in muscle mass during disuse conditions. Further investigation of the upstream signals governing Tfam expression in muscle is warranted. Supported by NSERC.
Chronic tadalafil treatment ameliorates functional muscle ischemia and exercise‐induced muscle injury in dystrophindeficient mdx mice
Authors: Li Liang , Zepeda Nancy , Victor Ronald G , Thomas Gail D
Published: June 2021

The dystrophin‐deficient muscles of patients with Duchenne or Becker muscular dystrophy and mdx mice are susceptible to ischemia during exercise due to loss of sarcolemmal neuronal nitric oxide synthase (nNOS). We showed that functional muscle ischemia is alleviated in patients and mice by acute treatment with the phosphodiesterase 5A (PDE5A) inhibitor tadalafil to boost NO‐cGMP signaling in the diseased muscles. We now asked if this anti‐ischemic effect is sustained during chronic PDE5A inhibition. We fed mdx mice control or medicated diets (tadalafil, 4 mg/kg) for 3 months and then evaluated norepinephrine (NE)‐induced hindlimb vasoconstriction. NE evoked similar decreases in femoral vascular conductance (FVC) in resting and contracting hindlimbs of untreated mdx mice, indicating functional muscle ischemia (ΔFVC contraction/rest, 1.07 ± 0.13; n=10). NE‐induced ischemia was attenuated in tadalafil‐treated mice (ΔFVC contraction/rest, 0.61 ± 0.06; n=9; P<0.05 vs untreated) and was similar to C57BL10 controls (ΔFVC contraction/rest, 0.50 ± 0.08; n=10). Serum creatine kinase activity was elevated 6‐fold post‐exercise in untreated mice, but only 2.5‐fold in treated mice (P<0.05). These findings indicate that chronic PDE5A inhibition counteracts functional muscle ischemia in mdx mice, which may reduce injury of the vulnerable dystrophin‐deficient muscles during exercise.

Supported by MDA, 158944.
Temporal effects of a severe‐heat load on intestinal integrity
Authors: Pearce Sarah C. , Sanz‐Fernandez Maria Victoria , Baumgard Lance H. , Gabler Nicholas K.
Published: June 2021

Acute heat stress (HS) and heat‐stroke can be detrimental to health and well‐being of humans and animals. Objectives were to chronologically characterize a severe heat‐load's impact on intestinal integrity. Crossbred gilts (n=40; 65 ± 2.7 kg BW) were subjected to HS conditions (37°C; 40% humidity) with ad‐libitum intake for either 0, 2, 4, 6, or 12 h (n=8/time point). Rectal temperature (Tr) and respiration rates (RR) were determined every 2 h. Pigs were euthanized at each time point and fresh ileum and colon samples were mounted into modified Ussing chambers. Transepithelial electrical resistance (TER) and fluorescein isothiocyanate (FITC)‐labeled dextran permeability (APP) were assessed. Additionally, ileum heat shock protein (HSP) 70 protein expression was determined. As expected, Tr increased linearly over time (P < 0.01) with the highest body temperature difference observed at 12 h (2.6°C). Compared to the 0 h group, RR increased (230%; P < 0.01) in the first 2 h and remained elevated during the 12 h (P <0.05). Ileal TER quadratically decreased (P < 0.01), reaching its minimum at 4h (40% decrease relative to 0h). Dextran APP quadratically increased with time (P < 0.05) peaking at 4h (1200%). Colon TER and dextran APP changed due to HS in quadratic responses over time (P = 0.05) similar to the ileal measurements, but were less pronounced. The stress marker HSP 70 increased in a linear fashion over time due to HS (P < 0.05). These data indicate that severe heat compromises intestinal integrity within 2 h and the maximum deleterious effects occur at 4 h.
Mechanism of replication‐coupled DNA interstrand cross‐link repair
Authors: Walter Johannes
Published: June 2021

Interstrand cross‐links (ICLs) are extremely toxic DNA lesions since they prevent any processes that require strand separation including DNA replication and transcription. In higher eukaryotes, the repair of these lesions takes place mainly in S‐phase, suggesting a connection to DNA replication. Many years of predominantly genetic research has shown that ICL repair requires the action of several classes of enzymes, including translesion DNA polymerases, structure‐specific endonucleases, recombinases, and numerous Fanconi anemia (FA) proteins. Using a Xenopus egg extract‐based ICL repair assay, we showed that lesion bypass during ICL repair occurs in several discrete steps that are coupled to DNA replication. We will present our latest results on how the FA proteins, BRCA1, and the recombination machinery promote replication‐coupled ICL repair.
Microbiota and glucocorticoids regulate postnatal developmental of intestinal Fut2 gene expression
Authors: Nanthakumar Nanda N , Meng Di , Newburg David S.
Published: June 2021

Intestinal maturation is regulated by intrinsic (glucocorticoid) and extrinsic (resident microflora) factors and the goal of this study is to determine the relative contribution of these two regulators in the postnatal development of α1,2‐fucosyltransferase II (Fut2) and sucrase‐isomaltase (SI) gene expression. In conventional raised (CONV) mice, ontogeny of Fut2 and SI gene expression was completed by fourth week. In the absence of colonization in germ‐free (GF) and bacteria‐depleted (BD) mice, intestinal Fut2 expression returned to low suckling levels. Ten days after colonization with adult, but not with suckling, gut microbiome, expression of Fut2 mRNA, α1,2‐fucosyltransferase activity and fucose α1,2‐linked glycoconjugates expression is fully recovered. In the GF mice, the rate of Fut2 and SI ontogeny was significantly delayed. Cortisone acetate (CA) precociously accelerated both Fut2 and SI gene expression in conventionally raised suckling pups. In the GF suckling mice, CA treatment precociously induced development of SI but not Fut2 gene expression. Thus, both microbes and glucocorticoids regulate gut ontogeny, but only suckling gut responds to CA and Fut2 expression remains primarily sensitive to adult microbiome. Complete recovery from mucosal injury caused by DSS‐treatment was observed in the CONV, but not in BD mice. Lack of recovery in BD mice correlated with the level of Fut2 expression in the gut. Thus, colonization by adult microbiota stimulated mature Fut2 gene expression and fucosylated gut phenotype; this colonization‐dependent Fut2 induction is necessary for full recovery from intestinal injury. This may explain the significant association observed between individuals expressing low Fut2 gene expression because of specific Fut2 polymorphic alleles and Crohn's disease and necrotizing enterocolitis.

Supported by D059126, HD013021, AI075563, and DK070260.
COMT rs4680 variant and cardiometabolic side‐effects in children treated with second‐generation antipsychotics
Authors: Cote Anita T , Panagiotopoulos Constadina , Devlin Angela M
Published: June 2021

Second‐generation antipsychotics (SGA) are used to treat children for a wide‐range of mental health conditions but are associated with side effects such as rapid weight gain; increased waist circumference; and elevated fasting plasma glucose, blood pressure, and metabolic syndrome (MetS). The aim of this study is to investigate the interaction of the rs4680 variant in the catechol‐ O‐methyltransferase gene (COMT) and SGA treatment on cardiometabolic side‐effects in a cross‐sectional population of SGA‐treated (n=99) and SGA–naïve (n=114) children. SGA‐treated children had higher (P<0.05) BMI z‐scores, systolic blood pressure (SBP) z‐scores, and fasting plasma glucose and total and LDL‐cholesterol concentrations than SGA‐naïve children. Fifteen percent of SGA‐treated children had MetS compared to 2% of SGA‐naïve children (P=0.001). The COMT rs4680 variant genotype frequencies were not different between SGA‐treated (AA 24.2%, AG 50.5%, GG 25.3%) and SGA‐naïve (AA 24.6%, GA 45.6%, GG 29.8%) children. An interaction between SGA‐treatment and COMT rs4680 genotype was observed such that SGA‐treated children with the AA genotype had higher SBP, fasting plasma insulin, and HOMA‐IR than those with the GG genotype. The opposite effect was observed in SGA‐naïve children. These findings suggest that the COMT rs4680 variant may interact with SGAs and contribute to cardiometabolic side‐effects in children.
The New Anatomist: Preparing to Teach in an Integrated Medical Curriculum.
Authors: Sorrentino Gina M.
Published: June 2021

The recent trend of curriculum integration in undergraduate medical education has resulted in a wide range of curricular designs. Many new graduates in the anatomical sciences receive their doctorate degrees from research‐centered institutions, and the teaching methods of the schools to which they are applying for faculty positions may be quite different and require significant adjustment. With proper guidance, graduates of the anatomical sciences can not only ease their transition into a new institution, but prepare themselves before graduation to take advantage of the job market. The author presents a personal account of the transition from a research‐centered PhD program in Anatomical Sciences to a fully integrated curriculum at a new medical school. Topics discussed here, with relevance to an integrated curriculum in particular, include: understanding the philosophies of the curriculum, understanding the role of anatomy and the anatomy professor, and creating opportunities for educational research in your new institution. Real life examples of how the missions and values of an integrated institution influence teaching are given. This account is aimed at anatomists who are about to graduate or have just graduated, and may be used a guide for degree granting programs in preparing their students for the variety of positions available.

Grant Funding Source: N/A
Addressing common and specific educational goals of the medical illustration discipline‐based anatomical courses that partially overlap with system‐based modules in the medical school curriculum
Authors: Stefan Cristian , Andrews William M , Harrison Steven J
Published: June 2021

The positive current trend of integrated curricula in most medical schools has resulted in fading borders between traditional disciplines. This situation offers extremely valuable opportunities as well as logistic challenges for programs in which students participate in selected curricular components. The junior students in our Medical Illustration (MI) Graduate Program take the Human Gross Anatomy and Neuroanatomy courses with the first‐year medical students. There are multiple advantages posed by this interaction. However, the MI discipline‐based courses have to follow the timeframes of the system‐based modules of the medical school curriculum. This situation raises aspects regarding: (a) scheduling, (b) overall content, (c) addressing common and specific goals and needs for each program, (d) providing the MI students with additional sessions and experiences that focus on creativity, critical thinking, and visual problem‐solving abilities, and (e) preparing appropriate exams. Our strategies in the academic years 2011–2013 have focused on the early shift from descriptive anatomy to deep learning based on functional, radiological and clinical applications to increasingly prepare the MI professionals for working with concepts, as their main task is to illustrate the complexity of the body, always with respect and reverence for the beauty and diversity of the human form in health and sickness.
Effect of Time Limits during Anatomy Practical Exams on Student Performance
Authors: Zhang Guiyun , Fenderson Bruce A , Schmidt Richard R , Veloski J Jon
Published: June 2021

We tested the hypothesis that timed anatomy practical exams are inherently more difficult than untimed exams. Students in the DPT program were assessed on their understanding of anatomic relationships using multiple‐choice questions. For the class of 2012 (n=46), students were allowed to circulate freely among 40 testing stations during the 40‐minute testing session. For the class of 2013 (n=46), students were required to move sequentially through the 40 testing stations (one minute per item). Students in both years were given three practical exams. An identical set of questions was used for both groups of students (untimed and timed exams). Our results indicate that there is no significant difference between student performance on untimed and timed exams. This result also held true for students in the top and bottom 20% of the class. Moreover, time limits did not lead to errors on even the most difficult, higher‐order questions. Thus, limiting time at testing stations during an anatomy practical exam does not adversely affect student performance. We believe that timed, multiple‐choice exams provide efficient and valid method available for assessing student understanding of anatomy.
Student perceptions and preferences of two TBL modalities in the second year medical curriculum
Authors: Waggoner Jane Suzanne , Braun Mark W. , O'Loughlin Valerie Dean
Published: June 2021

Since 2002, team‐based learning (TBL) has been an increasing component of US medical curricula. While numerous studies have examined TBL efficacy in medical curricula, little research has been done comparing differences in TBL modality (implementation and design).

This study examines the impact of TBL modality on 2nd year medical students’ perceived learning at Indiana University School of Medicine, Bloomington. Students participated in both medicine TBLs (traditional, standardized TBLs that used assigned groups and graded IRATs and GRATs) and pathology TBLs (nontraditional TBLs that used self‐selected groups and did not have an IRAT/GRAT). At the end of the academic year, students completed an anonymous survey comparing and contrasting the two TBL modalities. The survey contained both quantifiable Likert‐scale questions and open‐ended (qualitative) written comments. The written comments were examined for common theme, using a grounded theory approach.

Results showed that students preferred the non‐traditional TBLs to the standardized TBLs. In addition students felt the non‐traditional TBL approach allowed for a greater understanding of material and operated in a more efficient manner. Our research indicates TBL design and implementation impact student perceptions of this curricular tool, which should be taken into account in future TBL design and evaluation.
AN INTRODUCTION TO ULTRASONOGRAPHY: A NOVEL TEACHING SESSION FOR FINAL YEAR MEDICAL STUDENTS
Authors: Subbu Raj , Mackay Nicola , Abrahams Peter
Published: June 2021

The use of ultrasonography is increasingly being brought into clinical practice with certain procedures being carried out under ultrasound guidance; for example the insertion of central lines and chest drains. An understating of basic anatomy is essential to perform these procedures safely. The West Midlands Surgical Training Centre has a large collection of plastinated anatomy prosections which are used to improve anatomical understanding. The purpose of this session was to combine anatomy teaching and ultrasonography in a clinical setting. This teaching session was based around five separate stations focussing on anatomy of the neck, upper and lower limbs and consolidating this knowledge by identifying relevant structures using an ultrasound scanner. The clinical component was achieved with clinical examinations and procedures, for example obtaining an arterial blood gas sample. The interpretation and management of abnormal arterial blood gas results was then assessed. In this way we are able to illustrate the relevance of applying clinical anatomy, the use of ultrasonography, practical procedures and basic science as a combined set of skills essential for a junior doctor. The session is run for twenty students three times during the academic year. None of the students had any previous formal teaching in using ultrasonography. All students stated the set‐up of this teaching session highlighted the importance of combining clinical and basic science knowledge. This approach gives students the opportunity to improve their anatomy knowledge and develop skills using ultrasonography whilst maintaining its clinical importance.
3D Reconstructions of Early Heart Development as an Educational Resource
Authors: Piet Valerie Marie , Cork Robert John
Published: June 2021

Three‐dimensional reconstructions are a particularly useful tool for the visualization and understanding of complex subjects like embryo development. Early heart development is one such area. Our work has produced 3D reconstructions of human embryos from Carnegie stages 8, 9 and 10. Working with transverse serial‐sections provided by the Virtual Human Embryo databases we have used the computer programs Amira and Winsurf to produce our 3D reconstructions. Animations generated from these reconstructions are available in pdf form, or online, and can be incorporated into teaching material.

The Carnegie staging system divides the embryonic period into 23 stages based on the appearance of notable characteristics. Stages 8, 9, and 10 correspond roughly to 17–23 days of development. During this time period several heart development milestones occur. Beginning at stage 8, angioblastic clusters exist in horseshoe shaped cardiac mesoderm at the cranial end of the embryo. During stage 9, a pericardial cavity is present and by stage 10, an endocardial plexus is coalescing within the myocardium. Our reconstructions include the cardiogenic mesoderm, blood vessel primordia, endothelial tubes, myocardium, epicardium, and doral aorta.

The focus on early heart development allows us to visualize the important steps of forming the endothelial tubes and the relative position of the heart primordium in the embryo.

Grant Funding Source: NIH HD37811
Teaching Medicine through Media: An Interprofessional and Interdisciplinary Activity with Neuroscience and Film
Authors: Poznanski Ann , Afonso Nelia , Edwards Kyle , Vaughan Hunter , Bulgarelli Nancy , Krug Ernest , Gould Doug
Published: June 2021

Increasingly, the goal of medical training is to cultivate a well‐rounded professional with skills in both technical and humanistic realms. Toward this end, Oakland University William Beaumont School of Medicine (OUWB) faculty members have joined with colleagues at Oakland University (OU) for an interprofessional activity, which is part of our Neuroscience course. The learning experience involves a: Neuroanatomist, Medical Ethicist, Primary Care physician and Cinema Studies faculty at OU. The goal is to explore scientific, clinical, moral and aesthetic aspects of stroke by screening and discussing the film “The Diving Bell and the Butterfly”. Second year OUWB medical students and OU undergraduates in the Neurology Club, began with a reading assignment on “locked in syndrome”. Subsequently, the film was screened, then followed by a discussion with participants. This contextual, integrative learning has benefits for students, including: improved understanding of pathophysiological mechanisms and the moral dimensions of medicine and increased motivation and satisfaction; benefits for participating faculty include enhanced reflection on the moral, ethical and cultural aspects of medical practice, while liberal arts faculty gain exposure to the scientific basis of neural damage. The effectiveness of the activity was evaluated through participant assessment captured using an exit survey.
Active learning Cycle of Anatomy Laboratory for the Net Generation
Authors: De Leon Elizabel , Veras Wilson R
Published: June 2021

In America currently there are about 88 million members of the Net Generation (N‐Geners). The generation X (past generation) conducted their business mostly with the telephone and television. Today the “N‐Geners” not only has access to those devices but also super‐realistic video games, the internet, e‐mail, instant messaging, online communities, and videos and music that can be downloaded over a computer. This Generation is optimistic, positive, and driven to succeed. High achievers crave rewards and accolades for their hard work. They know that there are many significant problems affecting the world. They also know that the solutions to these problems are located in the youthful innovation and ever‐improving technology. Since 2010, we have been focused in developing a series of innovative strategies geared toward improving our teaching process in the Anatomy Laboratory in order to promote life long learning through interpersonal training and collaboration. They included dissection videos, Inatomy (muscuoskeletal application for classic Ipod), Ianatomy (Ipod Touch), Ipad iniative, virtual dissection, Virtual Reality, osteology, Imagenology, prosecction, plastination models, anatomical variation, superficial anatomy, muck simulation, self assessment, proctoring to proctor, and reciprocal peer teaching in the context of promoting spirituality, and professionalism, emphasizing the deep respect and gratitude for the human body that is made available to train the future generation of humanistic physicians.
MedStart: A Hands‐on Anatomical Experience for Middle School Students
Authors: Laitman Benjamin M , Witkin Ariana , Parikh Amar , Armstrong Anne , Zuckerman Samantha , Bhattacharya Romt , Chu Edward , Schneiderman Melissa , Palermo Ann‐Gel , Factor Stephanie H , Reidenberg Joy S , Laitman Jeffrey T
Published: June 2021

The goal of MedStart, a medical student run, week‐long science education summer camp, is to provide East Harlem middle school students with an appreciation of cardiovascular, pulmonary, and gastrointestinal system disease pathogenesis, treatment and prevention. Introducing students to the anatomical basis of health and disease is a vital component of our program. MedStart takes an interactive approach to education, dividing the camp up into five segments—vital signs, pulmonary, gastrointestinal, cardiovascular, and CPR/first aid—with strong hands‐on components during the organ system‐based days. In the summer of 2012 our program conducted a series of activities in which campers were able to examine manually normal and pathologic human cadaveric hearts, lungs, stomachs, intestines, and livers, in small group sessions led by Mount Sinai School of Medicine medical and graduate students. Campers were given assessments before and after the camp, which, in previous summers, have demonstrated increased knowledge gains in the organ systems we focused on. Along with its other hands‐on activities, MedStart is working to increase health knowledge and empower East Harlem youth to take control of their own health.

Grant Funding Source: American Association of Anatomists Educational Outreach Grant