bluuebunny/crossref_metadata_2025 huggingface dataset:
D‐glucose and D‐galactose affect Band 3 protein function and oxidative stress in human erythrocytes
Authors: Morabito Rossana , Remigante Alessia , Spinelli Sara , Cordaro Marika , Dossena Silvia , Marino Angela
Published: April 2020

Metabolic dysfunctions continuously expose human erythrocytes to sugars present in plasma during their circulatory life span of 120 days. Hyperglycemia in diabetes mellitus causes glycation of proteins along with oxidative stress in erythrocytes. Moreover, acute systemic administration of D‐galactose was used to artificially induce blood senescence in human models as well to mimic galactosemia. The purpose of this study is to understand the effect of D‐glucose and D‐galactose in human erythrocytes and in particular on the function of Band 3 protein, which is an excellent tool for studying the oxidative events on the erythrocyte membrane. Human erythrocytes in vitro were exposed to increasing concentrations of D‐glucose (5‐15‐35 mM) for 3h or D‐galactose (0.1‐3‐5‐10 mM) for 1h. The rate constant for SO4= uptake, accounting for the efficiency of anion exchange through Band 3 protein along with levels of TBARS levels and membrane –SH groups have been measured. When erythrocytes were exposed to high D‐glucose (15–35 mM) for 3 h, TBARS levels and membrane –SH groups were unchanged while the rate constant for SO4= uptake was significantly accelerated with respect to control (5 mM). After exposure to high D‐galactose (5–10 mM) for 1 h, TBARS levels were higher and membrane –SH levels were lower than control, while the rate constant for SO4= uptake was significantly slower with respect to control (0.1 mM). The present findings show that the measurement of the rate constant for SO4= uptake is a suitable tool to monitor the effect of sugars on erythrocytes; hyperglycemia due to high glucose applied for 3 h affects anion exchange capability without producing lipid peroxidation and oxidative protein damage. High concentrations of D‐galactose (> 5 mM) decreased the rate constant for SO4= uptake, induced lipid peroxidation and reduced total sulfhydryl content. Based on this evidence, future studies will evaluate if the acceleration of Band 3 protein in presence of high D‐glucose depends on altered Bp3 conformation, affecting crosslink with Hb, or on altered glycosylation or phosphorylation signaling underlying Band 3 protein function. In addition, alterations of parameters of oxidative stress caused by D‐galactose on the erythrocytes membrane open the view to consider a potential therapeutic strategy ameliorating the damage caused by D‐galactose.
Myoendothelial Junctions of Mature Coronary Vessels Express Notch Signaling Proteins
Authors: McCallinhart Patricia E. , Wani Elsa , Biwer Lauren A. , Wolpe Abigail G. , Isakson Brant E. , Lilly Brenda , Trask Aaron J.
Published: April 2020

The Notch signaling pathway, via cell‐cell communication, is critical during normal vascular development and homeostasis, and it is known to regulate vascular smooth muscle (VSMC) phenotype, including proliferation; however, few studies have addressed the role of Notch signaling in the mature vasculature. The current study tested the hypothesis that Notch signaling is present and active at mature myoendothelial junctions (MEJs), signaling hubs that occur at breaks in the internal elastic lamina (IEL). In mature, intact normal adult mouse coronary resistance microvessels (CRMs) embedded within heart sections, we observed Notch1, Notch2, Notch3, and Jagged1 expression within the fenestrae of the CRM IEL where MEJs are known to occur. Immunofluorescence clearly shows bridging from the VSMCs to endothelial cells (ECs). Immunofluorescence in larger mouse left anterior descending (LAD) coronary arteries demonstrate the expression Notch2 and Notch3 protein immunofluorescence within fenestrae of the IEL. We found that, on average, 39% and 33% of IEL fenestrae were positive for Notch2 and Notch3 expression, respectively. We also confirmed the presence of Notch3 expression in the MEJ by immunogold electron microscopy. In normal mouse femoral arteries, we found that, on average, 46%, 51%, and 58% of IEL fenestrae were positive for Notch1, Notch3, and Jagged1 expression, respectively. To determine whether Notch signaling components were localized to the MEJ, we co‐cultured primary human coronary ECs (hcECs) and primary human coronary VSMCs (hcVSMCs) across transwell inserts, and we utilized fluorescent microscopy to immunostain for Pai‐1, Jagged1, Notch1, and Notch2. PAI‐1 immunofluorescence was observed on the EC side of the transwells and within the transwell pores, indicating the formation of in vitro MEJs. We also identified distinct staining of Notch1, Notch2, and Jagged1 in the pores of the co‐culture transwells, which co‐localized with PAI‐1 within the pores. To assess whether Notch signaling was active at the MEJ, we co‐cultured hcECs and hcVSMCs on transwells to create in vitro MEJs. Smooth muscle cells were first transfected with a Notch‐sensor luciferase plasmid followed by transwell co‐culture of the transfected cells with endothelial cells. Notch activity was stimulated by 3‐fold across the MEJ in hcVSMC (1.00 ± 0.15 vs. 3.02 ± 0.43, p<0.001), which was completely abrogated by the non‐selective Notch γ‐secretase inhibitor, DAPT (1.15 ± 0.16, p<0.01). In this study, we demonstrate for the first time that Notch receptors and ligands are expressed within and are active at mature coronary MEJs, demonstrating a previously unrecognized mode of Notch signaling regulation between the endothelium and smooth muscle.

Support or Funding Information

Funding: EW was supported by NIH R25HD086885. NIH R00HL116769, R21EB026518, and S10OD023438 to AJT.
Lipoatrophy‐associated insulin resistance and hepatic steatosis are attenuated by intake of diet rich in omega 3 fatty acids.
Authors: Festuccia William T. , Moustaid-Moussa Naima , Moreira Rafael
Published: April 2020

We evaluated in this study glucose homeostasis and progression of nonalcoholic fatty liver disease (NAFLD) and hepatomegaly in severe lipoatrophic mice and their modulation by intake of a diet rich in omega 3 (n‐3) fatty acids. Severe lipoatrophic mice induced by adipocyte‐specific PPARγ deletion (A‐PPARγ KO) and littermate controls (A‐PPARγ WT) were evaluated for glucose homeostasis and liver mass, proteomics, lipidomics, inflammation and fibrosis after intake of a chow diet. Lipoatrophic mice were heavier than controls due to increased food intake and energy efficiency, severely glucose intolerant and hyperinsulinemic, and developed NAFLD characterized by increased liver glycogen, triacylglycerol and diacylglycerol contents, DNA and mitotic index, apoptosis, inflammation, steatosis score and fibrosis. This was associated with activation of de novo lipogenesis and liver enrichment with monounsaturated in detriment of polyunsaturated fatty acids including n‐3 fatty acids. Administration of a high‐fat diet rich in n‐3 fatty acids (HFO) inhibited de novo lipogenesis, enriched liver with n‐3 fatty acids, reduced hepatic steatosis, apoptosis, inflammation and steatosis and improved glucose homeostasis in lipoatrophic mice. In conclusion, diet enrichment with n‐3 fatty acids is an effective strategy to improve glucose homeostasis and reduce liver steatosis and inflammation without affecting hepatomegaly in severe lipoatrophic mice.

Support or Funding Information

Funded by FAPESP (2015/19530‐5).
Quantitatively defining spatial patterns of myoendothelial junctions in the arterial wall
Authors: Ruddiman Claire A. , Corliss Bruce A. , Peirce Shayn M. , Isakson Brant E.
Published: April 2020

The regulation of vasoconstriction and vasodilatory pathways is essential in resistance arteries, and can be modulated by heterocellular communication between the endothelial and smooth muscle cells. This can occur via direct endothelial‐smooth muscle cell contact at myoendothelial junctions (MEJs) through fenestrations in the internal elastic lamina (IEL). While it has been demonstrated that MEJs do not occupy every fenestration, their spatial distribution throughout the endothelium has not yet been defined and could reveal localization tendencies to cellular landmarks. We hypothesized that the MEJ and IEL fenestration spatial distribution would exhibit a non‐random distribution pattern throughout the endothelium with respect to various cellular landmarks, including the interendothelial junctions. In order to quantify this relationship, the spatial distributions of both holes and MEJs with respect to various cellular landmarks were examined. Third order mesenteric resistance arteries isolated from C57Bl6 male mice were prepared en face to view the endothelium and detect the IEL, nuclei, myoendothelial junctions, and interendothelial junctions (e.g., claudin‐5). A Matlab based analysis pipeline was developed to calculate the proximity of IEL fenestrations to cellular landmarks, such as interendothelial junctions. En face views of the IEL often have patches of varying signal intensity, posing a challenge for detecting fenestrations through basic thresholding methods. To overcome this, parameters within Image Processing Toolbox documentation were optimized for each image prior to thresholding. This allowed for accurate assignment of XY coordinates to fenestration boundaries (bwboundaries documentation) and calculation of center points. En face views of claudin‐5 were similarly thresholded and their boundaries also defined by XY coordinate matrix. Next, Euclidean distance was calculated between each fenestration center point and all claudin‐5 XY coordinates and plotted as a probability distribution. Monte Carlo simulations were also performed as a negative control. In the simulation, the fenestrations are replaced by randomly generated circles that mimic the number and radii of fenestrations in the original image. This is compared to the original distribution to assess if identified patterns are random. Our analysis of >100 simulations indicate that IEL fenestrations appear to be randomly distributed across the IEL. However, we find the MEJs within the IEL fenestrations may be strategically placed near interendothelial junctions. Preliminary data indicates that this particular MEJ localization pattern may be opportunistic because of the cytoskeletal components at interendothelial junction that can be recruited to support the signaling microdomains within MEJs.

Support or Funding Information

This work supported by 1F31HL149228‐01 (CAR) and HL088554 (BEI).
Influence of heat acclimation on spontaneous wheel‐run exercise in mice
Authors: Masuda Yuta , Marui Shuri , Kato Issei , Nagashima Kei
Published: April 2020

Introduction We previously found that, in mice, running distance on a running wheel went down in heat in mice, during which body core temperature became lower than that in no‐heat control. These results may suggest that mice determine exercise intensity depending on thermal input from the body surface. In the present study, we hypothesized that heat acclimation disinhibit such suppression of spontaneous running in heat.

Methods

Twenty two male C57BL/6J3 mice (age, 7 w) were used, which were divided to two groups: the heat acclimation and control groups (HA group and CON groups, respectively). They had surgery of intra‐abdominal placement of the IC‐chip sensor device (nanotag, Kissei, Nagano) for the measurement of abdominal temperature (Tabd) and counts of movement under inhalation anesthesia with sevoflurane and were allowed to recover for 10 d at least. They were individually housed at an ambient temperature (Ta) of 33°C and 25°C in the HA and CON groups, respectively for 2 w. Before and after the acclimation period, each mouse had two different trials on two‐separated days: exposures at 33°C and 25°C at 19:00–1:00 in a dark phase. In the trials, each mouse was moved to a semi‐closed chamber with a running wheel inside, where Tabd, running distance on the wheel, and metabolism by indirect calorimetry. After the experiments, mice were killed by overdose injection of pentobarbital, and the dorsal root ganglia (DRG, Th 5‐12) was obtained to assess protein expression of TRPV1 and V4.

Results

Before the acclimation period, the running distance was smaller in the in the 33°C trial than in the 25°C trial in both groups (P<0.05; e.g. 8.98 ± 0.86 and 13.68 ± 1.18 km, respectively, in the HA group). However, after the acclimation period, the distance in the HA group at 33°C trial became greater than the value before the acclimation period (P<0.05; 14.73 ± 1.32 km; post‐25°C, 14.50 ± 0.66 km).VO2 in the HA groups was higher in pre‐33°C trial and lower in post‐33°C trial than any other trial. Before the acclimation period, Tabd was lower in the 33°C trial than in the 25°C trial. After the acclimation period, Tabd was greater in the 33°C trial than in the 25°C trial. (pre‐33°C, 37.52 ± 0.28; pre‐25°C, 37.99 ± 0.30°C; post‐33°C, 37.93 ± 0.23; post‐25°C, 37.82 ± 0.30 °C). The expression TRPV1 on DRG tended to be lower in the HA group compared with the CON
group.

Conclusions

We found that, in mice, heat acclimation increases running distance on the wheel and decrease metabolic heat production. Based on physiological and biochemical analyses, decrease in thermal input from the skin and improve metabolic efficiency may be involved in the
mechanism.
The Role of PVN MC4R Neural Pathway in Autonomic Control of Metabolic Homeostasis and Cardiovascular Function
Authors: Singh Uday , Saito Kenji , Cui Huxing
Published: April 2020

Central melanocortin pathways, mainly via acting on melanocortin‐4 receptor (MC4R), play an important role in both energy balance and sympathetic traffics affecting cardiovascular function. MC4R deficiency in both humans and rodents causes obesity, insulin resistance and diabetes while maintaining low to normal sympathetic tone and blood pressure, suggesting a critical role of MC4R signaling in metabolic homeostasis and obesity‐associated sympathetic overdrive and hypertension. MC4R is widely expressed in the brain including hypothalamic paraventricular nucleus (PVN) which functions as an integrative center for energy metabolism and autonomic regulation. However, whether and how PVNMC4R pathways contribute to the sympathetic control of cardiovascular function and metabolic homeostasis remain incompletely understood. In present study, we first comprehensively mapped afferent projections of PVNMC4R neurons by stereotaxically delivering Cre‐dependent AAV driving expression of eYFP into the PVN of MC4R‐Cre knock‐in mice, which allow targeted anterograde tract‐tracing of PVNMC4R neurons throughout the brain. In addition to known brain region for feeding behavior (i.e. parabrachial nucleus), we found broad innervations of PVNMC4R neurons to various brain regions important for autonomic‐cardiovascular control and energy metabolism, including but not limited to nucleus of solitary tractus , dorsal motor nucleus of vagus, rostral ventrolateral medulla (RVLM) and spinal cord. Using a combination of sophisticated viral tools, we further reveal that thoracic spinal cord‐projecting PVNMC4R neurons are exclusively located in the posterolateral PVN and send collateral projections to RVLM. Functionally, microinjection of synthetic MC4R agonist (MTII) into the PVN evokes renal sympathetic nerve activity in anesthetized mice and chemogenetic activation of PVNMC4R neurons elevates blood pressure and decreases food intake in freely moving animals. Additionally, genetic ablation of PVNMC4R neurons in adult mice using Cre‐dependent AAV expressing Casp3 causes hyperphagia and massive weight gain (over 80% increase within 6 weeks). These results provide an important insight into the role of PVNMCR neural circuits in regulating metabolic homeostasis and cardiovascular function. Optogenetic and chemogenetic studies are ongoing to functionally dissect the neural circuits downstream of PVNMC4R neurons.

Support or Funding Information

National Institute of Health (R01), AHA Postdoctoral Grant Award (19POST34450083)
Instructional Hematology for Pre‐Health Undergraduates
Authors: Ogrodzinski Yvonne , Zubek John
Published: April 2020

At Michigan State University (MSU), the physiology department’s lab course for pre‐health professionals places an emphasis on laboratory exercises relevant to healthcare and the development of data analysis, professional development, and problem solving skills. As such, the goal of the hematology laboratory experience is to help students understand the ripple effect of hematology and how it can be extended to other fields of medicine. This is done by exposing students to the collection of blood samples and related laboratory tests that can be a typical part of a health screening or onsite wellness visit. Even for those students who do not become hematologists, it is important to maintain current knowledge of the best practices regarding the treatment and testing required for the best patient service and understand how various blood disorders affect certain populations.

We promote hematology learning through five blood testing stations in addition to microscope analysis. The five blood tests performed are hematocrit, blood typing, total lipid panel, prothrombin time and international normalized ratio (PT‐INR), and hemoglobin. Students will perform each of these blood tests with their own samples and are guided to the interpretation of each measurement with provided clinical standards. We also include the microscopic analysis of five live patient samples of blood with conditions including, sickle cell anemia, eosinophilia, acute myelogenous leukemia, and lymphoma. In analyzing each of these tests, students are tasked with relating their own data to overall health and clinical relevance. The learning objectives associated with these tests include the demonstration of safe handling of blood using Universal Precautions, among the ability to describe and recognize typical and pathologic values found in common blood tests. They are also required to explore the supporting physiology, such as the phases of hemostasis and consequences of incompatible blood transfusions. Students are able to practice their professional skills by teaching each other how to perform these tests after they have watched and performed their own, modeling the “see one, do one, teach one” pedagogy often found in clinical settings. Additionally, with the results and permission of our students, we are able to gather baseline blood data for this healthy age group and seek to compile and share these values as a composite look at typical data for this specific group.
Strategies for Mining Metagenomic Markers of the Gestational Diabetes Mellitus Microbiome
Authors: Bible Paul W.
Published: April 2020

Whole metagenome sequencing of pregnant mothers’ gut flora captures a snapshot of the microbial and metabolic landscape. Using case and control cohorts of pregnant Chinese women (n=60, 30‐30), metagenomic sequencing and clinical health data were mined using a simple correlation‐based coverage binning strategy to investigate microbiome features of gestational diabetes mellitus (GDM). Our approach sought to uncover novel associations between the hosts’ clinical variables of interest and specific genome regions of the resident gut bacteria. Candidate species were selected using two sample non‐parametric tests to determine differentially abundant species from sequencing. Next, available genomic reference sequences for the selected species were subdivided into 500 bp (base pairs) length sequence bins. Each bin is represented by the number of mapped sequences that overlap the location. After normalizing for depth of sequencing, each 500 bp region was assessed for its Spearman correlation with clinical variables across all samples. This method identified regions of interest that highly correlated with the results of the oral glucose tolerance tests after one hour (OGTT‐1). Using genome feature annotations of the reference sequence to identify nearby genes, our method identified the Starch‐binding protein SusD as an associated bacterial gene. Later, validation by PCR confirmed its differential abundance in GDM mothers compared to control pregnancies. These findings are to be published in an upcoming article; however, the mining strategy itself has not been fully investigates. In this work we compare the simple binning‐correlation strategy with more sophisticated de novo assembly followed by gene catalog construction and mapping. We find that the binning strategy is highly effective offering a simple but powerful data mining tool that compares favorably to more sophisticated and computationally costly methods.
Expression of Mitochondrial Complex 1 Inhibitor in Bovine Liver Tissue, Primary Hepatic Cells and Detection of its Transcript in Conditioned Media Mimicking Fatty Liver Disease
Authors: Choudhary Shanti , Lacasse Michelle , Choudhary Ratan K. , Rincon Mercedes , Beitz Donald C. , Testroet Eric D.
Published: April 2020

Mitochondrial complex I inhibitor (iC1) is a methylation‐controlled J protein that decreases oxidative phosphorylation of cellular respiration. Recent rodent studies showed loss or inhibition of iC1 was associated with prevention of lipid accumulation and T‐cell survival. One of the common metabolic disorder of dairy cattle is fatty liver disease (FLD), which often occurs during the periparturient period. Thus far, the evidence of iC1expression in bovine liver and its possible role in FLD development are not known. We hypothesized that iC1 expresses in bovine liver and associated with FLD during the periparturient period. We collected bovine liver tissue samples from abattoir and isolated primary hepatic cells using a simple nonperfusion‐based method. Primary hepatic cells were grown in presence of 0.4 mM palmitate, 20 ng/mL TNFα, and a cocktail of 0.4 mM palmitate and 20 ng/mL TNFα . The control cells grown in complete growth media (DMEM + 10% FBS +1x antibiotics) with no added treatments. For the first time, we 1) showed in vivo and in vitro expression of iC1 in bovine liver and 2) detected presence of iC1 specific transcript in aliquots of conditioned media of hepatic cells. Results of RT‐qPCR, immunocytochemistry and capillary‐based western blot showed expression of iC1 in bovine liver tissue and primary hepatic cells. Preliminary results of RT‐qPCR showed iC1 specific transcript amplification in conditioned media collected 24 h after treatments. Since the abundance of total RNA in conditioned media was lower, in comparison to the RNA from cell lysates, transcripts of iC1 was detectable at higher threshold cycle. The transcript of iC1 tended to be higher (4‐fold; p>0.05) in TNFα‐treated conditioned media vs. the control. Detection of iC1 transcript in hepatic cells and conditioned media is novel and may suggests iC1 expression by hepatic cells. Altogether, we showed expression of mitochondrial complex 1 inhibitor gene and protein in bovine liver tissue, primary hepatic cells and the conditioned media. Altering expression of iC1 to mitigate the metabolic disorder of fatty liver disease in periparturient dairy cows might be a noteworthy.
A role for Angiotensin‐(1‐7) and its receptors in sprouting angiogenesis and vascular homeostasis
Authors: Grant Gary , Bernadoni Juliana Lech , Soh Hong Jie , Walther Thomas
Published: April 2020

Background

Angiotensin converting enzyme 2 converts angiotensin (Ang) II into the biologically active heptapeptide Ang‐(1‐7). We have previously shown that Ang‐(1‐7) is involved in the stimulation and homeostasis of circulating endothelial progenitor cells, and that Ang‐(1‐7) is an endogenous ligand for the G protein‐coupled receptors Mas and MrgD. To better understand the role of the peptide in endothelial cell biology and homeostasis and the receptors involved, we aimed to investigate the effect of Ang‐(1‐7) on various functional parameters of endothelial cells.

Methods and Results

Ang‐(1‐7) was just as effective as vascular endothelial growth factor (VEGF) at stimulating endothelial tube formation in an in‐vitro Matrigel based assay in HUVEC. To investigate the effects in a more physiological model, we isolated aortas from Mas, MrgD, and Mas/MrgD deficient animals together with age and gender matched controls in an ex‐vivo model of sprouting. While aortas with a deficiency in either Mas or MrgD still showed normal number and length of sprouts as wild type mice, aortic rings deficient for Mas/MrgD showed a reduction in sprout length by 87% and no of sprouts by 77% when compared with wild type controls. We were also able to show that Mas deficient mice had a reduction in circulating CD34+ cells, a marker for heamopoeitic stem cells. Finally, we were able to show that after shunting blood flow to the hind limb of mice in an in‐vivo hind limb ischemia assay, Ang‐(1‐7) was able to restore 62.2% of blood flow in comparison with 17.1% in solvent‐treated mice.

Conclusion

All together, our data suggest a key role for Ang‐(1‐7) and its receptors in sprouting angiogenesis, vascular homeostasis, and thus might be a promising new option for the treatment of peripheral vascular disease.
Investigating the Function of Co‐activator‐associated Arginine Methyltransferase 1 (CARM1) During Denervation‐induced Skeletal Muscle Atrophy
Authors: Stouth Derek William , vanLieshout Tiffany , Ljubicic Vladimir
Published: April 2020

CARM1 is a type I protein arginine methyltransferase (PRMT) that forms asymmetric dimethylarginines (ADMA) by transferring monomethyl groups to arginine residues on target proteins. We previously observed elevated CARM1 expression and methyltransferase activity in response to neurogenic muscle disuse. Transient knockdown of CARM1 in muscle also represses the progression of muscle wasting and the expression of atrophy‐related genes following denervation. Notably, CARM1 regulates intracellular signaling molecules that are important for atrophy such as AMP‐activated protein kinase (AMPK) and downstream unc‐51 like autophagy activating kinase 1 (ULK1), forkhead box class O (FOXO), and transcription factor EB (TFEB) in non‐muscle tissues. The purpose of this study was to more comprehensively elucidate the function of CARM1 during neurogenic muscle disuse. We employed the Cre/loxP system to generate skeletal muscle‐specific CARM1 knockout (mKO) mice. Following unilateral sectioning of the sciatic nerve, mKO and wild type (WT) mice were subjected to 3 and 7 days of denervation. The contralateral limb served as an internal control. Hematoxylin and eosin staining analyses were performed to assess cross‐sectional area (CSA) in the extensor digitorum longus (EDL) muscle. We also employed Western blot and immunoprecipitation analyses to evaluate protein expression and protein interaction levels, respectively, in the non‐denervated and denervated tibialis anterior muscles. Our CSA analyses revealed that the absence of CARM1 mitigated denervation‐induced atrophy in the EDL muscle after 7 days of disuse (p < 0.05). Compared to WT animals, phosphorylation levels of AMPK and ULK1 were lower (p < 0.05) in mKO mice following 3 days of neurogenic muscle disuse. We also report that CARM1 interacts with AMPK, FOXO1, and TFEB in skeletal muscle. Relative to the contralateral, non‐denervated control (CON) limb, the interactions between CARM1‐AMPK and CARM1‐FOXO1 increased (p < 0.05) after 3 days of denervation in WT mice. When compared to the CON limb, the CARM1‐TFEB interaction was lower (p < 0.05) in WT mice following 7 days of neurogenic muscle disuse. In comparison to WT animals, the methylation of AMPK was significantly blunted in mKO mice after 3 days of denervation, suggesting that CARM1 methyltransferase activity regulates AMPK signaling. Collectively, these results suggest that CARM1 interacts with intracellular signaling molecules that are integral for muscle wasting. Importantly, targeting the interplay between CARM1‐AMPK, CARM1‐FOXO1, and CARM1‐TFEB may provide a novel therapeutic strategy for mitigating neurogenic skeletal muscle atrophy.

Support or Funding Information

Natural Sciences and Engineering Research Council of Canada

Canada Research Chairs
Dissect or not to dissect? Easy solution
Authors: Stashenko Vetaley
Published: April 2020

Throughout the recorded past (from Edwin Smith Surgical Papyrus) anatomy education did not experience dramatic changes. Practically until the last 5–10 years we were using only a text and a cadaver. Only did generation Z start to experience an intervention of AI and VR in our dissection rooms and lecture halls. Literature research exhibits convincing evidence of the positive impact of these new pedagogical tools on student’s success and overall performance (Freeman et al. 2014; Michael 2006). Some new methods of active learning, however, have been proven to be more effective than others. Lombardi et al. (2014) observed that students who used plastic models, rather than prosected specimens or virtual dissections, scored considerably higher on both initial and follow‐up exams despite student perceptions that the different forms of actual dissections were of more value. Fancovicova et al. (2014) observed that students who had admittance to multiple forms of active learning performed at a higher level than students who had access to only one form (including dissection). Herur et al. (2011) reported that students who utilized multiple forms of active learning had significantly higher levels of retention at 15 and 30 days after the class than those who learned by passive means.

There is a perception that dissection continues to be recommended as the golden standard of anatomical education. Our presentation will present our experience of exploration of the possibilities to improve student’s success through the creation of a technologically advanced anatomy classroom that allows the learning anatomy of living human beings, not only cadavers. First‐time students are able to hold their own 3D printed organs during lab exercises. We can observe anatomical variations and their effects on living specimens in real time. At the same time, traditional cadaver dissections allow us to appreciate the complexity of the human body.
Meta‐Analysis Reveals Gut Microbial Signatures in Colorectal Adenoma and A Link with Secondary Bile Acid Conversion
Authors: Wu Yuanqi , Jiao Na , Wu Dingfeng , Fang Sa , Gao Wenxing , He Xiaosheng , Zhu Lixin , Zhu Ruixin
Published: April 2020

Background

Colorectal adenoma is a common stage during the colorectal cancer (CRC) development. The early diagnosis during adenoma stage could be a key to effectively reduce CRC. Here, we examined the microbial biomarkers of adenoma aiming to identify non‐invasive early diagnosis markers for the prevention of CRC.

Design

We performed a meta‐analysis of four publicly available 16S rDNA datasets with 307 colorectal adenomas, 217 CRC and 252 control samples. Microbial compositions and functions were identified with DADA2 and PICRUSt2 in QIIME2. After adjusting confounding factors, differential analysis was conducted to determine the unbiased significant differential microbes and then Random Forest model was constructed to identify biomarkers. Finally, we applied internal and external validation to assess the diagnostic capabilities with these biomarkers.

Results

43 differential microbes between adenoma and control were observed. After adjusting the potential confounding factors, 11 of these microbes were identified as biomarkers that could distinguish adenoma from healthy status, with a robust diagnostic accuracy (AUC=0.80). Internal and external validation confirmed the robustness and consistent diagnostic capabilities of these biomarkers with an average AUC of 0.72. Meanwhile, we identified 117 differential microbes between adenoma and CRC. Among them, 26 microbes may serve as biomarkers to separate adenoma from CRC (AUC=0.8), with an average validation AUC of 0.77. Eubacterium ruminantium group was the only one species that appeared in both sets of biomarkers. In addition, multiple microbial functions were altered in microbiome, including secondary bile acid conversion as well as lipopolysaccharide metabolism.

Conclusion

Meta‐analysis strongly supports that microbial markers can distinguish among CRC, colorectal adenoma and healthy controls and therefore serve as non‐invasive screening tools for the early diagnosis and treatment of CRC. CRC and adenoma specific alterations in the microbial composition and microbial metabolic pathways provide potential targets for the prevention and intervention of CRC.

Support or Funding Information

This work was supported by National Natural Science Foundation of China 81774152 (to RZ) and 81770571 (to LZ).
Comprehensive analysis of policies of whole body donor programs within the United States
Authors: Elzie Carrie , Cruz Bianca
Published: April 2020

Many schools around the United States use cadavers in their instruction; however, the acquisition of these bodies vary immensely from school to school. Previous studies have shown that many schools around the United States lack detail in their consent process. Therefore, we looked at all US whole body bequeathal programs (state and school specific) to assess the donor programs’ processes of selecting and receiving donors. Criteria reviewed included consent for education, consent for research, bequeathal policies, weight limit, reasons for refusal, time limit after death, and other policies. Programs of interest included all full body donation programs within the United States that were affiliated with or donated to colleges/universities. A total of 114 programs were analyzed.

Programs that were no longer accepting donations were omitted. Information was gathered from published brochures and donation forms, as well as, direct inquiry with programs. Any information that could not be found through the consent forms or on the website was obtained by placing a call to the office. This was repeated for each donation program and the results were analyzed to compare the process, extent, and development of cadaver lab donation programs.

All programs provided consent forms that specifically included the use for education. Consent for research was documented in 102 (89%) of the programs. For statewide programs, donors could request a specific school, although their desire was not guaranteed. In every program, donors were able to fill out donation forms in advance. In addition to individual bequeathal, many programs also allow donation by the next of kin. The majority of programs (55%) had a weight and or a BMI limit with the average of 240 pounds. Forty‐four (39%) programs had no weight limit but denied donors based on a BMI above 30. 5. Only 4% programs had no weight requirement and would evaluate the donors’ eligibility once the body was received. The most commonly cited reasons for refusal were autopsy, infectious disease, recent surgery, trauma, and malnourishment/obesity. Costs incurred to donate ranged from being free of cost to $750. At time of death, most facilities had a regimen of when they needed to receive the donation. All facilities requested to be notified as soon as possible and preferred receipt of the body anywhere between two and 168 hours from the time of death.

A review of the donor programs within the US revealed mostly consistencies across policies. However, the biggest variable was related to cost. Future studies should look at the amount of donations each program receives and determine correlations between the aforementioned policies. Looking into ways to streamline the donation process and procedures would make it easier for potential donors to maximize their options and for all involved to be informed.
Metabolic responses to intermittent hypoxia are regulated by sex and gonadal hormones in mice.
Authors: Marcouiller François , Jochmans-Lemoine Alexandra , Ganouna Gauthier , Mouchiroud Mathilde , Laplante Mathieu , Marette André , Joseph Vincent
Published: April 2020

Sleep apnea is a respiratory disease characterized by intermittent hypoxia (IH) due to repeated obstructions of the airways during sleep. Men and women after menopause are more likely to develop sleep apnea than women before menopause. In addition to disrupting sleep quality, sleep apnea is strongly associated with metabolic syndrome and type 2 diabetes, and the metabolic consequences of sleep apnea may vary according to sex or the hormonal status. We therefore used intact males, intact females, and ovariectomized (OVX) female mice to ask if IH induces sex‐specific metabolic responses and if endogenous female hormones prevent these effects. Within each group, mice were exposed to room air (RA) or IH (nadir 6% O2, 10 cycles/hour, 12 hours/day) during 2 weeks. Body weights were measured before and after the exposures to IH. At the end of the IH exposures, mice were fasted for 6 hours, a blood sample was drawn from the saphenous vein to measure fasting plasma glucose and insulin levels. We then performed intraperitoneal insulin or glucose tolerance tests to further assess glucose homeostasis. In distinct groups, we assessed liver levels of glycogen, triglycerides (TG), and expression levels of selected genes involved in glucose metabolism. In intact males, IH reduced weight gain, lowered the fasting levels of glucose and insulin in plasma, and slightly increased glucose tolerance. In intact females, the only effect of IH was a modest decrease of glucose tolerance. In OVX females, IH exposures reduced fasting glucose and insulin levels, induced insulin resistance, and led to a major glucose intolerance. In the liver, IH exposures slightly increased glycogen levels in males and OVX females and had a profound effect to decrease TG levels only in males. The expression levels of key liver glycolytic (LDH) and mitochondrial (Citrate synthase, PDH, PDK) genes were reduced by IH in males or in OVX females. We conclude that the metabolic consequences of IH exposures are sex specific. In females, endogenous ovarian hormones appear to prevent the development of a major glucose intolerance during IH exposure and help maintaining a normal expression pattern of hepatic genes involved in glucose metabolism. Surprisingly, the effects of IH are different in males and OVX females, suggesting that endogenous testosterone might also have a role in males.

Support or Funding Information

Supported by CIHR.
Longitudinal analyses of epigenetic pathways in deployed active duty Service Members
Authors: Gautam Aarti , Yang Ruoting , Chakraborty Nabarun , Abu-Amara Duna , Hammamieh Rasha , Marmar Charles , Jett Marti
Published: April 2020

Background

Pre‐deployment identification of risk/ resilience factors is crucial in developing strategies to reduce or prevent post‐traumatic stress (PTSD) symptoms in military personnel. Management of PTSD is complicated because of the overlapping symptoms with the comorbid conditions and the diagnostic reliance on self‐report and time consuming psychological evaluation. A more comprehensive understanding of molecular pathophysiology of PTSD could facilitate an unbiased biomarker‐driven next‐generation intervention strategy. Herein, we study the epigenomic consequences of combat elicited PTSD in a longitudinal manner using blood methylation analysis.

Methods

Blood samples were collected pre‐deployment, immediately post‐deployment, and at a three month follow‐up. This study assessed participants prior to stressor exposure during the index deployment and included comprehensive whole blood analysis. We collected self‐reports such as PTSD Checklist for DSM‐5 (PCL‐5) along with clinical tests. DNA methylation assays were carried out using Illumina 450K , followed by repeated‐measures ANOVA, correlation analysis, and PTSD differential analysis to characterize methylation status alterations associated with significant PTSD Check List (PCL) changes (PCL difference >10).

Results

We identified 64 subjects with changes in PCL scores of 10 across the three phases of the sample collection. The correlation analysis between DNA methylation and the PCL5 trend identified genes from T cell pathways, dopamine receptor signaling and fatty acid activation pathways. We observed shifts in DNA methylation that distinguish PTSD from controls and also that tracks PTSD as measured by PCL5.

Conclusions

We observed shifts in DNA methylation related to deployment related adverse health outcomes. Active Duty cohort represented a real‐world scenario measured stress before and a short time after combat‐related trauma. Future work will integrate these epigenetic data with other molecular and clinical findings to better understand PTSD co‐morbidities, as well as susceptibility and resiliency.

Support or Funding Information

Research was conducted in compliance with all Federal requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army.
Effect of concentration and staining duration of iodine contrast applied to study the masticatory muscle of Wistar rats in micro‐CT images
Authors: Piau Fabio Giardiello , Haddad Juliana , Rossi Ana Cláudia , Prado Felippe Bevilacqua , Freire Alexandre Rodrigues
Published: April 2020

The challenge to study the anatomy of the muscular system is to replace destructive methods such as macroscopic dissection with non‐destructive methods involving magnetic resonance imaging and computed microtomography (micro‐CT). The aim of this study was to evaluate the effect of concentration and staining duration of iodine contrast applied to study the masticatory muscle of Wistar rats in micro‐CT images. Fifteen male Wistar rats (Rattus norvegicus albinus), 2 months old (200–250g) from CEMIB‐UNICAMP were used. After euthanasia, the animals were randomly assigned to the experiments in 3 groups of 5 rats each, with the iodine contrast solutions of 3%, 5% and 10%. Each group was subdivided into 3 subgroups according to the staining duration: 7 days, 15 days and 30 days. The pieces were scanned on a 1174 Skyscan microtomograph (Bruker, Belgium) with a voltage of 50 Kv and amperage of 800 μA at the Piracicaba Dental School‐UNICAMP. The scanning of the pieces was done with a pixel dimension equal to 31 μm. After scanning, the images were exported to the software NRecon Reconstruction (Bruker, Belgium) in which they were reconstructed three‐dimensionally in axial sections presenting the x‐ray attenuation coefficients with values relative to the muscle structure reinforced by contrast. The reconstructed images were exported to the software Materialise MIMICS v18 (Materialise, Belgium) to evaluate the anatomical characteristics as well as the density of masticatory muscles. The muscles observed were: Temporal, Masseter (superficial part), Masseter (deep part), External pterygoid, Internal pterygoid and Zigomaticomandibular. These parameters were evaluated in each piece to compare the effects of different contrast concentration and staining duration. The 5% concentration showed better characterization of muscle groups, regardless of the period. At 5%, the 30‐day period showed better definition of muscle fibers. The 3% concentration presented only muscle mass and its contours, but there was no definition in the image that could differentiate each muscle. In the group with a concentration of 10%, contrast enhancement was observed only on the muscular surfaces. Thus, the solution concentration was the most effective parameter to stain the muscle fibers. The preparation and application protocol of iodine‐based contrast solution was most effective at 5% concentration with 30 days action.

Support or Funding Information

National Council for Scientific and Technological Development (PIBIC/CNPq).
High plasma levels of exosomal miR21 and miR133a are associated with LV reverse remodelling after surgical mitral valve repair
Authors: Furini Giulia , Pizzino Fausto , Casieri Valentina , Mariani Massimiliano , Solinas Marco , Maffei Stefano , Chiappino Dante , Aquaro Giovanni D. , Lionetti Vincenzo
Published: April 2020

Mitral valve regurgitation (MR) is the most common cardiac valvular disease associated with increased morbidity and mortality. Patients are characterized by volume overload and progressive increase of left ventricular (LV) end‐diastolic volume (EDV). Surgery is recommended for patients with substantial loss of global cardiac function (LV ejection fraction, LVEF). A correctly timed surgery for MR can reverse LV remodeling. Identification of patients at high risk of post‐operative LV remodeling may help to act with preventive strategies. In this scenario, microRNAs delivered by plasma exosomes (pEXOs), smallest extracellular nanovesicles, might have a predictive value. Primary MR patients (N=19; 45–71 y.o.) underwent implantation of a prosthetic mitral ring. LV remodeling was assessed by cadiac magnetic resonance imaging and pEXOs were isolated by optimized ultracentrifugation before surgery (T0) and six months after surgery (T1). Isolated pEXOs were quantified by nanoparticle tracking analysis (NanoSight) and miR‐1, miR‐21, miR‐133a and miR‐208a were measured by RT‐qPCR. The same analysis was performed healthy subjects with normal cardiac function (Control, N=8). pEXOs levels at T0 were lower (−32%, p=0.02) in patients with worst postoperative LV function, while they were higher at T1 (+31%, p=0.03) in patients with LV reverse remodeling after surgery. At T1, the increase in pEXOs levels was associated to decreased heart mass index (−13%, p=0.02) and higher levels of exosomal miR‐21 (+78%, p=0.02) and miR‐133a (+69%, p=0.05) were detected in patients with improved LV function. In conclusion, higher postoperative levels of pEXOs and exosomal miR‐21 and 133a mark LV reverse remodeling after mitral valve repair. Combined measurements of circulating exosomes and their microRNAs cargo might represent potential novel perioperative markers in patients with MR.