Medicine and Health Sciences Commons^™

Late Summer Planted Oat-Brassica Forage Quality Changes During Winter Grazing, Mary E. Lenz, Jordan L. Cox, Kristin E. Hales Hales, Hannah C. Wilson, Mary E. Drewnoski

Nebraska Beef Cattle Reports

Oat, radish, and turnip samples were taken on pivots being grazed from November through January in Clay Center, Nebraska. The objective was to determine how the quality changed throughout the winter. The forage was observed to be high-quality (highly digestibility with moderate CP content). Digestibility did decline over this period but minimal changes in CP content were observed. From early November to early January, the digestibility of oats appeared to decline more (10% unit decline) than turnip and radish leaves (5% unit decrease). However, digestibility (67% IVOMD) and CP content (15%) of oats in early January were still as high …

Comparison Of Two Alternate Prostaglandin Products In Yearling Beef Heifers, Alicia C. Lansford, Tonya L. Meyer, Richard N. Funston Funston

Nebraska Beef Cattle Reports

Yearling heifers were administered 1 of 2 alternate prostaglandin products (Lutalyse vs. Lutalyse HighCon), which differ in concentration of active ingredient and administration route. Timing of estrus, pregnancy rate to AI, and final pregnancy rate did not differ between treatments. Body weight and ADG were also not affected by prostaglandin treatment. These results indicate producers can utilize Lutalyse HighCon, administered subcutaneously (s.c.), to avoid injection site blemishes and reduce carcass discounts with no impact on estrus synchronization or pregnancy rates.

Impact Of Syngenta Enogen Feed Corn On Finishing Cattle Performance And Carcass Characteristics, Melissa L. Jolly-Breithaupt, Curtis J. Bittner, F. Henry Hilscher Hilscher, Galen E. Erickson, James C. Macdonald, Matt K. Luebbe

Nebraska Beef Cattle Reports

Two finishing experiments were conducted to evaluate Syngenta Enogen Feed Corn containing an alpha amylase enzyme trait compared to the near negative isoline control corn at two locations on cattle performance and carcass characteristics. No statistical differences were observed for final BW, DMI, ADG, or F:G for steers fed Syngenta Enogen Feed Corn versus the near negative isoline control corn. Fat depth and calculated yield grade were greater for steers fed Syngenta Enogen Feed Corn; however, HCW and marbling scores were not different. Previous research has observed a 2.6% to 16.4% decrease in F:G when Syngenta Enogen Feed Corn was …

Chopped Sugar Beets As A Component Of Beef Cow Diets, Karla H. Jenkins, Jacob A. Hansen

Nebraska Beef Cattle Reports

Sugar beets not accepted for human consumption were evaluated as an energy source in a limit fed ration for confined beef cows. Initial and final body weight and body condition score were not different between wheat straw based diets containing wet distillers grains and either chopped sugar beets or corn. Body condition score change tended to be greater for the diet containing corn. However, cows on both treatments finished the study with a body condition score of 5.7. Chopped sugar beets mixed with wheat straw and stored in an agricultural bag underwent ensiling and did not result in choking issues. …

Evaluation Of Brown Midrib Corn Silage For Growing And Backgrounding Beef Steers, F. Henry Hilscher, Curtis J. Bittner, John N. Anderson Anderson, Galen E. Erickson

Nebraska Beef Cattle Reports

A growing study evaluated three corn silage hybrids for growing crossbred steers. The three hybrids were: a standard corn silage hybrid which served as the control, a brown midrib hybrid, and an experimental brown mid rib hybrid with a softer endosperm. Intake, ADG, and ending BW were greater for steers fed either brown mid rib silage compared to control, but not different between the brown mid rib or experimental brown mid rib silage. While brown mid rib hybrids had greater DMI and ADG, there was no difference in F:G between all three treatments. Feeding brown mid rib hybrids as corn …

Evaluation Of Corn Silage Hybrids With The Brown Midrib Trait And Silage Inclusion For Finishing Cattle, F. Henry Hilscher, Curtis J. Bittner, John N. Anderson Anderson, Galen E. Erickson

Nebraska Beef Cattle Reports

A finishing study evaluated three corn silage hybrids fed at either 15 or 45% of diet DM for finishing steers. The three hybrids were a standard corn silage hybrid which served as the control, a brown midrib hybrid and an experimental brown midrib hybrid with a softer endosperm. An interaction was observed between hybrid and silage inclusion. Gain and HCW were greater for steers fed the experimental brown midrib compared to other two hybrids when fed at 15%. Feeding brown midrib hybrids at 45% of the diet DM resulted in greater ADG and HCW when compared to a control corn …

Nutrient Digestibility And Fermentation Of Brown Midrib Corn Fed To Growing Beef Steers, F. Henry Hilscher, Melissa L. Jolly-Breithaupt, Jana L. Gramkow Gramkow, Hannah C. Wilson, Mitch M. Norman, John N. Anderson, Galen E. Erickson

Nebraska Beef Cattle Reports

A digestion study evaluated three corn silage hybrids for growing crossbred steers. The three hybrids were: a standard corn silage hybrid which served as the control, a brown midrib hybrid (bm3), and an experimental bm3 hybrid with a softer endosperm. Both bm3 hybrids had greater organic matter and fiber digestibility compared to the control corn silage. However, no differences were observed between the two bm3 hybrids. Rumen pH was reduced for BM3 and BM3-EXP compared to the control suggesting greater rumen fermentation. In vitro gas production was increased for the bm3 hybrids compared to the control further supporting greater rumen …

The Relationship Between Marbling, Superoxide Dismutase, And Beef Tenderness, Nicolas J. Herrera, Emery K. Wilkerson, Katherine I. Domenech-Perez Perez, Felipe A. Ribeiro, Michael D. Chao, Chris Calkins

Nebraska Beef Cattle Reports

This study was conducted to evaluate the relationships between animal oxidative status (as indicated by superoxide dismutase [SOD] activity) to marbling and beef tenderness. Prime and Select-grade strip loins were selected and aged for 2, 7, 14, 21, and 28 days for Warner Bratzler shear force, Troponin-T, and SOD activity. Results showed that meat exhibiting higher levels of marbling had lower shear force values and thus were more tender. Low-marbled samples tended to have a greater tenderness response to aging. The effect of oxidative stress, however, was not evident in this study as SOD values were similar. Although the effects …

Evaluation Of Corn Distillers Solubles On Finishing Steer Performance, Benjamin H. Hansen, Curtis J. Bittner, Bradley M. Boyd Boyd, F. Henry Hilscher, Jim C. Macdonald, Galen E. Erickson

Nebraska Beef Cattle Reports

A finishing study was conducted to evaluate the effects of feeding 0, 8, 16, or 20% corn distillers solubles (CDS), as well as the effects of feeding a combination of 16% CDS and 20% wet distillers grains plus solubles (WDGS) to replace a blend of dry-rolled and high-moisture corn on finishing steer performance. As inclusions of CDS increased, ADG linearly increased and F:G linearly decreased. Feeding value of CDS at 20% inclusion was determined to be 147% compared to the corn blend. The addition of WDGS resulted in a decrease in DMI with similar ADG, resulting in a decrease in …

Effect Of Myoglobin State On Color Stability Of High Pressure Processed Ground Beef, Jhinuk Gupta, Chad G. Bower, George A. Cavender Cavender, Gary A. Sullivan

Nebraska Beef Cattle Reports

High pressure processing, a non-thermal pasteurization technique, can reduce E. coli in beef but the use is limited due to discoloration of raw beef after high pressure processing. Different states of myoglobin have inherently different color stability. The objective of this study was to determine the impact of myoglobin state on color stability of raw beef patties treated with high pressure processing. Modified atmosphere packaging (high oxygen-oxymyoglobin, carbon monoxide-carboxymyoglobin), vacuum packaging (deoxymyoglobin) or added potassium ferricyanide (metmyoglobin) treatments were used to prepare patties with desired myoglobin states. Color was measured (CIE L*, a*, b*) before and after high pressure processing …

Effect Of Ingredients And Packaging On Color Of High Pressure Processed Ground Beef, Jhinuk Gupta, Chad G. Bower, George A. Cavender Cavender, Gary A. Sullivan

Nebraska Beef Cattle Reports

High pressure processing is a non-thermal pasteurization technique to control pathogens, like E. coli. However, color changes in raw beef induced by processing restrict high pressure processing’s use within the beef industry. The objectives of this study were to investigate the effects of adding curing agents (nitrite) and packaging with or without reducing compounds (ascorbic acid/erythorbate) on color retention in high pressure processed ground beef. High pressure processing resulted in a detrimental effect on the color of the beef patties for all treatments. Lightness and yellowness increased and redness decreased after high pressure processing. The effect remained the same throughout …

Effects Of Production System On Cow And Calf Performance, Shelby E. Gardine, Jason M. Warner, Robert G. Bondurant Bondurant, F. Henry Hilscher, Karla H. Jenkins, Galen E. Erickson, Terry J. Klopfenstein

Nebraska Beef Cattle Reports

Limited traditional forage resources have prompted interest for alternative cow-calf production systems. This study evaluated the effects of two winter cow-calf production systems (cornstalk grazing and dry-lot feeding) on cow-calf performance in a summer-calving, intensively managed cowherd at two locations. Grazing cow-calf pairs on cornstalks resulted in similar or lower ending BW of cows and lower ADG of calves when compared to cow-calf pairs wintered in the dry-lot. A partial budget of incorporating winter cornstalk grazing into an intensive production system suggests that cows wintered on cornstalks may be $137 more profitable compared to cows wintered in the dry-lot.

Effects Of Production System And Post-Weaning Management On Finishing Performance And Carcass Characteristics Of Steer And Heifer Calves, Shelby E. Gardine, Bradley M. Boyd, Curtis J. Bittner Bittner, F. Henry Hilscher, Karla H. Jenkins, Galen E. Erickson, Terry J. Klopfenstein

Nebraska Beef Cattle Reports

This study evaluated the effects of cow-calf production system and post-weaning management on finishing performance and carcass characteristics of steer and heifer calves. Calves that were wintered on cornstalks prior to weaning had lighter initial BW compared to calves that were wintered in the dry-lot; however, final BW and carcass weight were similar between treatments. Post-weaning management was either adapting calves to a finishing diet following weaning or feeding a grower diet prior to the finishing phase. When harvested at similar back fat, calves that were fed a grower diet for 76 days prior to the finishing phase had 71 …

Evaluation Of Protein From Distillers Grains In Finishing Diets On Nutrient Digestibility, Zachary E. Carlson, Jana L. Gramkow, Hannah C. Wilson Wilson, Melissa L. Jolly-Breithaupt, Galen E. Erickson, Jim C. Macdonald, Matt K. Luebbe

Nebraska Beef Cattle Reports

A metabolism trial was conducted to evaluate protein from modified distillers grains plus solubles (MDGS) in finishing diets on nutrient digestibility and ruminal fermentation characteristics. Isolated protein from corn was not different than MDGS for dry matter, organic matter, or neutral detergent fiber digestibility. However, steers fed MDGS tended to have lower total tract organic matter digestibility compared to corn and protein from corn. Protein had greater total tract organic matter and starch digestibility than MDGS. Protein from corn did not contribute towards the lower digestibility of MDGS. Protein is more easily digestible than the other components in distillers grains …

Production Of High-Quality Beef–The Nebraska Advantage, Chris R. Calkins

Nebraska Beef Cattle Reports

Here is a clear description of the Nebraska Advantage in producing high-quality beef: the right genetics, numerous pastures, a multitude of cattle, great water resources, locally-grown corn, the availability of ethanol byproducts for feed, and young cattle that grow fast, with sufficient marbling and subcutaneous fat to ensure tender, flavorful, beef. The science of beef quality supports this production system. The state supports its citizens and agriculture through the University of Nebraska and the Nebraska Department of Agriculture, augmenting the infrastructure. If one could invent an ideal place to produce high-quality beef, it would look much like Nebraska! The impact …

Potential Variation In Determination Of Longissimus Muscle Area In Carcasses From Heifers Fed With Or Without Zilpaterol Hydrochloride, Chris R. Calkins, Jenna J. O'Daniel, Govindarajan Konda Naganathan Naganathan, Kellen B. Hart, Katherine I. Domenech-Perez, T.B. Schmidt

Nebraska Beef Cattle Reports

This study was conducted to evaluate some sources of potential variation in determination of longissimus muscle area between the 12th and 13th ribs of carcasses from heifers fed with or without zilpaterol hydochloride. Cross sections of the rib-loin were taken cranial to the 11th rib and caudal to the 13th rib, about 3–4 millimeters thick at 90 degrees perpendicular to the long axis of the longissimus muscle. Potential variation of longissimus muscle area can arise from the natural variation of muscle size when cutting between the 12th and 13th ribs (up to 9%). Deviation in cutting perpendicular to the long …

Impact Of Feeding Distillers Grains With Or Without Oil Removal As Well As Supplemental Corn Oil On Nutrient Digestibility By Finishing Cattle, Jordan E. Burhoop, Melissa L. Jolly-Breithaupt, Jana L. Gramkow Gramkow, Matthew K. Luebbe, Jim C. Macdonald, Galen E. Erickson

Nebraska Beef Cattle Reports

A digestion trial was conducted to determine the effects of the removal of corn oil from modified distillers grains plus solubles (MDGS) and the impact of supplemental corn oil on finishing cattle nutrient digestion. Four treatments were evaluated: a corn control diet (CON), 40% de-oiled MDGS (DO MDGS), or 38% de-oiled MDGS plus 2% corn oil (MDGS + Oil) formulated to equal the fat content of FF MDGS, or 40% full fat MDGS (FF MDGS). Treatment differences were observed for digestibility of dry matter, organic matter, and fiber, but not for fat. When oil was added to de-oiled MDGS, digestibility …

Effects Of Late Gestation Supplementation, Synchronization, And Creep Feeding In A Spring Calving Beef Herd In The Nebraska Sandhills, Devin Broadhead, L. A. Stalker, Jacki A. Musgrave Musgrave, Richard N. Funston

Nebraska Beef Cattle Reports

Supplementation during winter grazing increased cow BW and BCS but had no effect on reproduction or calf performance, thus increasing production costs without increasing returns. Using a CIDR for estrus synchronization in a herd with existing acceptable reproductive performance did not increase cow pregnancy rate or advance calving date. Feeding creep feed to calves is an effective means of increasing weaning BW which resulted in increased live and carcass weight. Under the conditions of this study, when additional feed and price slide for heavier calves was considered, feeding creep feed did not result in added profits.

Field Evaluations Of Insecticide Modes Of Action Classes For Control Of Horn Flies In Nebraska, David J. Boxler, Gary J. Brewer, Richard N. Funston Funston, Jacqueline A. Musgrave

Nebraska Beef Cattle Reports

Insecticides of different Mode of Action (MoA) classes were tested for their ability to reduce horn fly populations on cattle in Nebraska pastures between 2009 and 2016. Macrocyclic lactone products were the most efficacious, reducing horn fly numbers by an average of 93% over ten location years of testing. Organophosphate and pyrethroid MoA products, tested in 7 and 12 location years, reduced fly numbers by 75% and 73%. Classes tested only once were METI (88% reduction) and a combination of organophosphate + pyrethroid (64%).

Evaluation Of Revalor-Xh For Beef Heifers Fed Different Days On Feed, Robert G. Bondurant, Marshall N. Streeter, Galen E. Erickson Erickson, James C. Macdonald, Matt K. Luebbe

Nebraska Beef Cattle Reports

Heifers were treated with either no implant, an initial implant of Revalor-200 and re-implanted with Revalor-200, or Revalor-XH and assigned to one of four serial slaughter harvests at 151, 165, 179, and 193 days on feed to determine the effects on feedlot performance and carcass characteristics. Implanting heifers increased final BW, ADG and HCW while decreasing marbling score and improving feed efficiency compared to non-implanted heifers. Increasing days on feed decreased ADG while increasing feed efficiency, HCW, fat thickness, marbling score, and calculated yield grade. By increasing HCW sold, implanting revenue can be maximized, assuming added risk for YG discounts.

Effect Of Supplementation During The Breeding Season On A May-Calving Herd In The Nebraska Sandhills, Alicia C. Lansford, Jacki A. Musgrave, Richard N. Funston Funston

Nebraska Beef Cattle Reports

Females in their first and second breeding season received either a 32% crude protein (DM) supplement or no supplement throughout the breeding season. Supplementation did not affect heifer BW, BCS, and pregnancy rate at pregnancy diagnosis. Supplementation impacted primiparous cow BW and BCS at pregnancy diagnosis, but did not affect pregnancy rates. Calf birth weight and dystocia rates were unaffected by supplementation for both heifers and primiparous cow. Calves nursing supplemented dams were heavier at weaning. Greater supplementation may be needed to affect pregnancy rate.

Sivcpz Closely Related To The Ancestral Hiv-1 Is Less Or Non-Pathogenic To Humans In A Hu-Blt Mouse Model, Zhe Yuan, Guobin Kang, Lance Daharsh, Wenjin Fan, Qingsheng Li

Nebraska Center for Virology: Faculty Publications

The HIV-1 pandemic is a consequence of the cross-species transmission of simian immunodeficiency virus in wild chimpanzees (SIVcpz) to humans. Our previous study demonstrated SIVcpz strains that are closely related to the ancestral viruses of HIV-1 groups M (SIVcpzMB897) and N (SIVcpzEK505) and two SIVcpz lineages that are not associated with any known HIV-1 infections in humans (SIVcpzMT145 and SIVcpzBF1167), all can readily infect and robustly replicate in the humanized-BLT mouse model of humans. However, the comparative pathogenicity of different SIVcpz strains remains unknown. Herein, we compared the pathogenicity of the above four SIVcpz strains with HIV-1 using humanized-BLT mice. …

Identification Of Unequally Represented Founder Viruses Among Tissues In Very Early Siv Rectal Transmission, Jian Chen, Yanqin Ren, Lance Daharsh, Lu Liu, Guobin Kang, Qingsheng Li, Qiang Wei, Yanmin Wu, Jianqing Xu

Nebraska Center for Virology: Faculty Publications

Characterizing the transmitted/founder (T/F) viruses of multi-variant SIV infection may shed new light on the understanding of mucosal transmission.We intrarectally inoculated six Chinese rhesus macaques with a single high dose of SIVmac251 (3.1 × 10⁴ TCID₅₀) and obtained 985 full-length env sequences from multiple tissues at 6 and 10 days post-infection by single genome amplification (SGA). All 6 monkeys were infected with a range of 2 to 8 T/F viruses and the dominant variants from the inoculum were still dominant in different tissues from each monkey. Interestingly, our data showed that a cluster of rare T/F viruses …

Analytical Performance Of A Low‐Cost Multiplex Polymerase Chain Reaction Human Papillomavirus Genotyping Assay For Use In Sub‐Saharan Africa, Kandali Samwel, Crispin Kahesa, Julius Mwaiselage, Daniela Gonzalez, John T. West, Charles Wood, Joel Palefsky, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

We have tested a multiplex polymerase chain reaction (PCR) human papillomavirus (HPV) genotyping assay to fill the need for rapid and low‐cost HPV detection in Sub‐ Saharan Africa. This method allows high throughput genotyping and simultaneous detection of 14 high‐risk and two low‐risk HPV types, by PCR amplification of HPV DNAs in a single reaction tube. In this study, we describe stepwise experiments to validate the multiplex HPV PCR assay for determination of HPV genotypes from 104 cervical brush samples from Tanzanian women. Assay performance was evaluated by determination of intra‐laboratory reproducibility, sensitivity, and specificity. Further performance was assessed by …

Discovery Of A Non-Nucleoside Rna Polymerase Inhibitor For Blocking Zika Virus Replication Through In Silico Screening, Aryamav Pattnaik, Nicholas Palermo, Bikash R. Sahoo, Zhe Yuan, Duoyi Hu, Arun S. Annamalai, Hiep L. X. Vu, Ignacio Correas, Pavan Kumar Prathipati, Christopher J. Destache, Qingsheng Li, Fernando A. Osorio, Asit K. Pattnaik, Shi-Hua Xiang

Nebraska Center for Virology: Faculty Publications

Zika virus (ZIKV), an emerging arbovirus, has become a major human health concern globally due to its association with congenital abnormalities and neurological diseases. Licensed vaccines or antivirals against ZIKV are currently unavailable. Here, by employing a structure-based approach targeting the ZIKV RNA-dependent RNA polymerase (RdRp), we conducted in silico screening of a library of 100,000 small molecules and tested the top ten lead compounds for their ability to inhibit the virus replication in cell-based in vitro assays. One compound, 3-chloro-N-[({4-[4-(2-thienylcarbonyl)-1-piperazinyl]phenyl}amino)carbonothioyl]-1-benzothiophene-2-carboxamide (TPB), potently inhibited ZIKV replication at submicromolar concentrations. Molecular docking analysis suggests that …

Impact Of Pre-And Postpartum Nutrition On March-Calving Cow And Progeny Productivity, Jacki A. Musgrave, Devin L. Broadhead, L. A. Stalker Stalker, Richard N. Funston

Nebraska Beef Cattle Reports

March-calving cows were fed hay or grazed corn residue prepartum, and then either fed hay or grazed subirrigated meadow postpartum. Combinations of these different feeding strategies impacted body weight (BW) and body condition score (BCS) during the pre-and postpartum period; however, resulted in similar pregnancy rates. Although differences were not observed in cow pregnancy rates, a benefit in pre-weaning growth was observed for calves from the dams in postpartum meadow treatment. A tendency for an improvement in marbling score was observed for steers born to cows fed hay prepartum, perhaps indicating a higher plane of nutrition prepartum may improve quality …

Efficacy Of Oral Administration Of Sodium Iodide To Prevent Bovine Respiratory Disease Complex, B. M. Shoemake, B. L. Vander Ley, B. W. Newcomer, M. C. Heller

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: The prevention of bovine respiratory disease complex (BRD) in beef cattle is important to maintaining health and productivity of calves in feeding operations.

Objective: Determine whether BRD bacterial and viral pathogens are susceptible to the lactoperoxidase/hydrogen peroxide/ iodide (LPO/H₂O₂/I^-) system in vitro and to determine whether the oral administration of sodium iodide (NaI) could achieve sufficient concentrations of iodine (I) in the respiratory secretions of weaned beef calves to inactivate these pathogens in vivo.

Animals: Sixteen weaned, apparently healthy, commercial beef calves from the University of Missouri, College of Veterinary Medicine teaching herd. …

Genes For Membrane Transport Proteins: Not So Rare In Viruses, Timo Greiner, Anna Moroni, James L. Van Etten, Gerhard Thiel

James Van Etten Publications

Some viruses have genes encoding proteins with membrane transport functions. It is unknown if these types of proteins are rare or are common in viruses. In particular, the evolutionary origin of some of the viral genes is obscure, where other viral proteins have homologs in prokaryotic and eukaryotic organisms. We searched virus genomes in databases looking for transmembrane proteins with possible transport function. This effort led to the detection of 18 different types of putative membrane transport proteins indicating that they are not a rarity in viral genomes. The most abundant proteins are K+ channels. Their predicted structures vary between …

Size-Dependent Catalysis Of Chlorovirus Population Growth By A Messy Feeding Predator, John Delong, Zeina Al-Ameeli, Shelby Lyon, James L. Van Etten, David Dunigan

James Van Etten Publications

Many chloroviruses replicate in endosymbiotic zoochlorellae that are protected from infection by their symbiotic host. To reach the high virus concentrations that often occur in natural systems, a mechanism is needed to release zoochlorellae from their hosts. We demonstrate that the ciliate predator Didinium nasutum foraging on zoochlorellae-bearing Paramecium bursaria can release live zoochlorellae from the ruptured prey cell that can then be infected by chloroviruses. The catalysis process is very effective, yielding roughly 95% of the theoretical infectious virus yield as determined by sonication of P. bursaria. Chlorovirus activation is more effective with smaller Didinia, as larger …

A Rapid, Handheld Device To Assess Respiratory Resistance: Clinical And Normative Evidence, Aaron B. Holley, Wesley D. Boose, Michael Perkins, Karen L. Sheikh, Nancy P. Solomon, Angela M. Dietsch, Jafar Vossoughi, Arthur T. Johnson, Jacob F. Collen

Department of Special Education and Communication Disorders: Faculty Publications

Introduction: Following reports of respiratory symptoms among service members returning from deployment to South West Asia (SWA), an expert panel recommended pre-deployment spirometry be used to assess disease burden. Unfortunately, testing with spirometry is high cost and time-consuming. The airflow perturbation device (APD) is a handheld monitor that rapidly measures respiratory resistance (APD-R_r) and has promising but limited clinical data. Its speed and portability make it ideally suited for large volume pre-deployment screening. We conducted a pilot study to assess APD performance characteristics and develop normative values.

Materials and Methods: We prospectively enrolled subjects and derived …