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9 ARTICLES PUBLISHED IN JoVE

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Biology

In vivo Liver Endocytosis Followed by Purification of Liver Cells by Liver Perfusion
Sandhya Gopalakrishnan 1, Edward N. Harris 1
1Department of Biochemistry, University of Nebraska, Lincoln

The study of liver sinusoidal endothelial cells (SECs) must be performed with primary cells obtained from the animal as no cell lines exist. This method relies on liver digestion and differential centrifugation for SEC purification for subsequent culturing and experimentation.

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Biology

Introducing an Angle Adjustable Cutting Box for Analyzing Slice Shear Force in Meat
Tom Whitesell 1, Carmen Avilés 2, Jennifer L. Aalhus 1, Chris R. Calkins 3, Ivy L. Larsen 1, Manuel Juárez 1
1Lacombe Research Centre, Agriculture and Agri-Food Canada, 2Grupo de investigación MERAGEM, Universidad de Córdoba, 3Department of Animal Science, University of Nebraska

Slice shear force is a reference method for beef texture analysis. Using an angle adjustable cutting box could increase its accuracy for research purposes. The results from different locations within the longissimus muscle show a high correlation with Warner-Bratzler shear force methodology and high potential adaptability for different muscles.

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Biochemistry

Purification of Hepatocytes and Sinusoidal Endothelial Cells from Mouse Liver Perfusion
Fatima Cabral 1, Colton M. Miller 1, Katrina M. Kudrna 1, Blake E. Hass 1, Jocelyn G. Daubendiek 1, Brianna M. Kellar 1, Edward N. Harris 1
1Department of Biochemistry, University of Nebraska

The goal of this protocol is to obtain high viability and high yield of hepatocytes and sinusoidal endothelial cells from liver. This is accomplished by perfusing the liver with a type IV collagenase solution via the portal vein, followed by differential centrifugation to obtain hepatocytes and sinusoidal endothelial cells.

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Developmental Biology

Isolation of Specific Neuron Populations from Roundworm Caenorhabditis elegans
Edward M. Germany 1, Nataliya Zahayko 1, Oleh Khalimonchuk 1,2,3,4
1Department of Biochemistry, University of Nebraska, 2Nebraska Redox Biology Center, University of Nebraska, 3Nebraska Center for Integrated Biomolecular Communication, University of Nebraska, 4Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center

Here, we present a protocol for simple isolation of specific groups of live neuronal cells expressing green fluorescent protein from transgenic Caenorhabditis elegans lines. This method enables a variety of ex vivo studies focused on specific neurons and has the capacity to isolate cells for further short-term culturing.

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Chemistry

Chromatographic Fingerprinting by Template Matching for Data Collected by Comprehensive Two-Dimensional Gas Chromatography
Federico Stilo 1, Chiara Cordero 1, Carlo Bicchi 1, Daniela Peroni 2, Qingping Tao 3, Stephen E. Reichenbach 3,4
1Dipartimento di Scienza e Tecnologia del Farmaco, Università degli Studi di Torino, 2SRA Instruments, 3GC Image LLC, 4Computer Science and Engineering Department, University of Nebraska

This protocol presents an approach to fingerprint and explore multi-dimensional data collected by comprehensive two-dimensional gas chromatography coupled to mass spectrometry. Dedicated pattern recognition algorithms (template matching) are applied to explore the chemical information encrypted in the extra-virgin olive oil volatile fraction (i.e., volatilome).

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Medicine

Utilizing Percutaneous Ventricular Assist Devices in Acute Myocardial Infarction Complicated by Cardiogenic Shock
Shuktika Nandkeolyar 1, Poonam Velagapudi 2, Mir B. Basir 3, Aditya S. Bharadwaj 1
1Division of Cardiology, Loma Linda University Medical Center, 2Division of Cardiology, University of Nebraska, 3Division of Cardiology, Henry Ford Health System

Percutaneous ventricular assist devices are increasingly being utilized in patients with acute myocardial infarction and cardiogenic shock. Herein, we discuss the mechanism of action and hemodynamic effects of such devices. We also review algorithms and best practices for the implantation, management and weaning of these complex devices.

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Medicine

The Intra-Aortic Balloon Pump
Ganesh Gajanan 1, Emmanouil S. Brilakis 2, Jolanta M. Siller-Matula 3,4,5, Ronald L. Zolty 1, Poonam Velagapudi 1
1Division of Cardiovascular Medicine, University of Nebraska Medical Center, 2Minneapolis Heart Institute, 3Department of Internal Medicine II, Division of Cardiology, Medical University of Vienna, 4Department of Experimental and Clinical Pharmacology, Medical University of Warsaw, Center for Preclinical Research and Technology CEPT, 5Department of Internal Medicine II, Division of Cardiology, Medical University of Vienna

We describe the steps for the percutaneous implantation of the intra-aortic balloon pump (IABP), a mechanical circulatory support device. It acts by counterpulsation, inflating at the onset of diastole, augmenting diastolic aortic pressure and improving coronary blood flow and systemic perfusion, and deflating before systole, reducing left ventricular afterload.

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Medicine

Use of a Percutaneous Ventricular Assist Device/Left Atrium to Femoral Artery Bypass System for Cardiogenic Shock
Swethika Sundaravel 1, Poonam Velagapudi 1, Mamas Mamas 2, Sandeep Nathan 3, Alexander Truesdell 4
1University of Nebraska Medical Center, 2Keele Cardiovascular Research Group, Keele University, 3University of Chicago Medicine, 4Virginia Heart/Inova Heart and Vascular Institute

The following article describes the stepwise procedure for placement of a device (e.g., Tandemheart) in cardiogenic shock (CS) that is a percutaneous left ventricular assist device (pLVAD) and a left atrial to femoral artery bypass (LAFAB) system that bypasses and supports the left ventricle (LV) in CS.

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Biochemistry

Recombinant Production of Bifidobacterial Endoglycosidases for N-glycan Release
Berfin Sucu 1, Ayşe Bayraktar 1,2, Hatice Duman 1, Ayşenur Arslan 1, Merve Kaplan 1, Melda Karyelioğlu 1, Eda Ntelitze 1, Taner Taştekin 1, Seray Yetkin 1, Melih Ertürk 2, Steven A. Frese 3,4, Bethany M. Henrick 4,5, Hacı Mehmet Kayili 6, Bekir Salih 7, Sercan Karav 1
1Department of Molecular Biology and Genetics, Çanakkale Onsekiz Mart University, 2Uluova Dairy Farm, 3Department of Nutrition, University of Nevada, 4Department of Food Science and Technology, University of Nebraska, 5Evolve BioSystems Inc., 6Department of Biomedical Engineering, Karabuk University, 7Department of Chemistry, Hacettepe University

Bifidobacteria possess a unique genomic capability for N-glycan cleavage. Recombinantly producing these enzymes would be a promising novel tool to release bioactive N-glycans from glycoprotein-rich substrates such as colostrum.

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