Life Sciences Commons^™

Evaluation Of A Field-Deployable Reverse Transcription-Insulated Isothermal Pcr For Rapid And Sensitive On-Site Detection Of Zika Virus, Mariano Carossino, Yanqiu Li, Pei-Yu A. Lee, Chuan-Fu Tsai, Pin-Hsing Chou, Dennis Williams, Ashley Skillman, R. Frank Cook, Grayson Brown, Hsiao-Fen G. Chang, Hwa-Tang T. Wang, Udeni B. R. Balasuriya

Maxwell H. Gluck Equine Research Center Faculty Publications

Background: The recent emergence of Zika virus (ZIKV) in Brazil and its precipitous expansion throughout the Americas has highlighted the urgent need for a rapid and reliable on-site diagnostic assay suitable for viral detection. Such point-of-need (PON), low-cost diagnostics are essential for ZIKV control in vulnerable areas with limited resources.

Methods: We developed and evaluated a ZIKV-specific field-deployable RT-iiPCR reagent set targeting the E gene for rapid detection of ZIKV in ZIKV-spiked human and mosquito specimens, and compared its performance to the Center for Disease Control and Prevention (CDC) and Pan American Health Organization (PAHO) RT-qPCR assays targeting the E …

The Pul37 Tegument Protein Guides Alphaherpesvirus Retrograde Axonal Transport To Promote Neuroinvasion, Alexsia L. Richards, Patricia J. Sollars, Jared D. Pitts, Austin M. Stults, Ekaterina E. Heldwein, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

A hallmark property of the neurotropic alpha-herpesvirinae is the dissemination of infection to sensory and autonomic ganglia of the peripheral nervous system following an initial exposure at mucosal surfaces. The peripheral ganglia serve as the latent virus reservoir and the source of recurrent infections such as cold sores (herpes simplex virus type I) and shingles (varicella zoster virus). However, the means by which these viruses routinely invade the nervous system is not fully understood. We report that an internal virion component, the pUL37 tegument protein, has a surface region that is an essential neuroinvasion effector. Mutation of this region rendered …

Zika Virus Encoding Nonglycosylated Envelope Protein Is Attenuated And Defective In Neuroinvasion, Arun Saravanakumar Annamalai, Aryamav Pattnaik, Bikash R. Sahoo, Ezhumalai Muthukrishnan, Sathish Kumar Natarajan, David Steffen, Hiep Vu, Gustavo A. Delhon, Fernando Osorio, Thomas M. Petro, Shi-Hua Xiang, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

ABSTRACT Zika virus (ZIKV), a mosquito-transmitted flavivirus responsible for sporadic outbreaks of mild and febrile illness in Africa and Asia, reemerged in the last decade causing serious human diseases, including microcephaly, congenital malformations, and Guillain-Barré syndrome. Although genomic and phylogenetic analyses suggest that genetic evolution may have led to the enhanced virulence of ZIKV, experimental evidence supporting the role of specific genetic changes in virulence is currently lacking. One sequence motif, VNDT, containing an N-linked glycosylation site in the envelope (E) protein, is polymorphic; it is absent in many of the African isolates but present in all isolates from the …

Avs Corner, December 2017, Farzana Ferdous (Advisor), Caitlin Mathews, Emily Alexander, Brooke Redmond, Meghan Courey, Erika Jones, Wendy Guzman, Peyton Voirin, Morgan Forte

AVS Corner

No abstract provided.

Zika Virus Encoding Nonglycosylated Envelope Protein Is Attenuated And Defective In Neuroinvasion, Arun S. Annamalai, Aryamav Pattnaik, Bikash R. Sahoo, Ezhumalai Muthukrishnan, Sathish Kumar Natarajan, David Steffen, Hiep Vu, Gustavo Delho, Fernando Osorio, Thomas M. Petro, Shi-Hua Xiang, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Zika virus (ZIKV), a mosquito-transmitted flavivirus responsible for sporadic outbreaks of mild and febrile illness in Africa and Asia, reemerged in the last decade causing serious human diseases, including microcephaly, congenital malformations, and Guillain-Barré syndrome. Although genomic and phylogenetic analyses suggest that genetic evolution may have led to the enhanced virulence of ZIKV, experimental evidence supporting the role of specific genetic changes in virulence is currently lacking. One sequence motif, VNDT, containing an N-linked glycosylation site in the envelope (E) protein, is polymorphic; it is absent in many of the African isolates but present in all isolates from the recent …

Effect Of Endophyte-Infected Tall Fescue Toxins On Growth Performance And The Microbial Community In The Rumen And Feces In Pregnant Ewes, Saleh Alrashedi

Graduate Theses and Dissertations

Fifty pregnant Katahdin ewes grazed pastures with high (HE) and moderate (MI) levels of Neotyphodium coenophialum-infected tall fescue between February 1 and May 24 to determine the impacts of tall fescue toxins on body weight (BW), body condition scores, FAMACHA scores, fecal egg counts (FEC), and serum Cu, Zn, prolactin, and non-esterified fatty acids (NEFA). Within those 50 ewes, 15 were selected randomly for fecal and rumen sampling to assess the impacts of HE and MI on the microbiome community. Total bacterial DNA was extracted from rumen and fecal samples, and alpha and beta diversities were used to relate microbiome …

Development Of An Animal Model For Enterovirus For Evaluation D68 For Screening Of Antiviral Therapies, W. Joseph Evans

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Enterovirus D68 (EV-D68) virus has become more prevalent over the last 15 to 20 years. EV-D68 attacks the respiratory system and can cause severe disease in individuals who have underlying respiratory problems. There have also been reports of individuals with EV-D68 showing signs of neurological system problems and acute flaccid paralysis. Because of the increase in patients with EV-D68 and also the potential for neurological disease, an animal model is needed to study the disease and to evaluate experimental therapies for EV-D68 infection.

To develop the animal model, 4-week old AG129 mice that lack alpha and beta interferon receptors, making …

Genetic Signatures For Helicobacter Pylori Strains Of West African Origin, Kennady K. Bullock, Carrie L. Shaffer, Andrew W. Brooks, Ousman Secka, Mark H. Forsyth, Mark S. Mcclain, Timothy L. Cover

Veterinary Science Faculty Publications

Helicobacter pylori is a genetically diverse bacterial species that colonizes the stomach in about half of the human population. Most persons colonized by H. pylori remain asymptomatic, but the presence of this organism is a risk factor for gastric cancer. Multiple populations and subpopulations of H. pylori with distinct geographic distributions are recognized. Genetic differences among these populations might be a factor underlying geographic variation in gastric cancer incidence. Relatively little is known about the genomic features of African H. pylori strains compared to other populations of strains. In this study, we first analyzed the genomes of …

Histomonas Elisa, Jessica L Martinez

EURēCA: Exhibition of Undergraduate Research and Creative Achievement

Histomonas meleagridis is a protozoan parasite of avians and is the causative agent in Histomonosis, commonly known as Blackhead Disease. Current methods for diagnosing the presence of H. meleagridis are limited to parasite culture or Polymerase Chain Reaction (PCR) to amplify target DNA. This project aims to develop an enzyme-linked immunosorbent assay (ELISA) for faster and more sensitive diagnosis of Histomonas infections. Cultures of H. meleagridis parasites were purified, and surface antigens were extracted using a spectrum of chemical solutions. The various antigen solutions were subjected to an ELISA, with serum from birds immunized for H. meleagridis as positive controls. …

Autonomic Nervous System Reactivity In A Free-Ranging Mammal: Effects Of Dominance Rank And Personality, Elodie F. Briefer, James A. Oxley, Alan G. Mcelligott

Alan G. McElligott, PhD

Modulation of the autonomic nervous system (ANS) activity allows animals to effectively respond to internal and external stimuli in everyday challenges via changes in, for example, heart and respiration rate. Various factors, ranging from social such as dominance rank to internal such as personality or affective states can impact animal physiology. Our knowledge of the combinatory effects of social and internal factors on ANS basal activity and reactivity, and of the importance that each factor has in determining physiological parameters, is limited, particularly in nonhuman, free-ranging animals. In this study, we tested the effects of dominance rank and personality (assessed …

Effectiveness Of Composting As A Biosecure Disposal Method For Porcine Epidemic Diarrhea Virus (Pedv)-Infected Pig Carcasses, Sarah Vitosh-Sillman, John Dustin Loy, Bruce Brodersen, Clayton Kelling, Kent M. Eskridge, Amy Millmier Schmidt

School of Veterinary and Biomedical Sciences: Faculty Publications

Porcine epidemic diarrhea virus (PEDV) is an enteric disease of swine that has emerged as a worldwide threat to swine herd health and production. Substantial research has been conducted to assess viability of the virus on surfaces of vehicles and equipment, in feed and water, and on production building surfaces, but little is known about the persistence in PEDV-infected carcasses and effective disposal methods thereof. This study was conducted to quantify the persistence of PEDV RNA via quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) at various time-temperature combinations and in infected piglet carcasses subjected to composting. Although this method …

Differential Roles For Inner Membrane Complex Proteins Across Toxoplasma Gondii And Sarcocystis Neurona Development, Rashmi Dubey, Brooke Harrison, Sriveny Dangoudoubiyam, Giulia Bandini, Katherine Cheng, Aziz Kosber, Carolina Agop-Nersesian, Daniel K. Howe, John Samuelson, David J. P. Ferguson, Marc-Jan Gubbels

Maxwell H. Gluck Equine Research Center Faculty Publications

The inner membrane complex (IMC) of apicomplexan parasites contains a network of intermediate filament-like proteins. The 14 alveolin domain-containing IMC proteins in Toxoplasma gondii fall into different groups defined by their distinct spatiotemporal dynamics during the internal budding process of tachyzoites. Here, we analyzed representatives of different IMC protein groups across all stages of the Toxoplasma life cycle and during Sarcocystis neurona asexual development. We found that across asexually dividing Toxoplasma stages, IMC7 is present exclusively in the mother’s cytoskeleton, whereas IMC1 and IMC3 are both present in mother and daughter cytoskeletons (IMC3 is strongly enriched in daughter buds). In …

Rehabilitation Of An Injured Golden Eagle (Aquila Chrysaetos): A Case Study, Fida Muhammad Khan, Fakhar-I- Abbas, Abu Ul Hassan Faiz

Journal of Bioresource Management

Habitat destruction and anthropogenic activities cause wild birds to migrate towards urban areas in search of food and sometimes nesting in high-roof buildings, where they are caught by local people using different techniques and are further used for hunting or recreational purposes. An injured Golden Eagle (Aquila chrysaetos) was found entangled in bushes near the vicinity of Balkasar Research Complex, Chakwal, Pakistan. After complete physical examination, a wound on the right wing and closed leg fracture was diagnosed. The successful treatment was done by anti-parasitic and anti-bacterial drugs with careful monitoring. Splint was adjusted after aligning the fractured …

Evaluation Of The Effect Of Serum Antibody Abundance Against Bovine Coronavirus On Bovine Coronavirus Shedding And Risk Of Respiratory Tract Disease In Beef Calves From Birth Through The First Five Weeks In A Feedlot, Aspen M. Workman, Larry A. Kuehn, Tara G. Mcdaneld, Michael L. Clawson, Carol G. Chitko-Mckown, John Dustin Loy

School of Veterinary and Biomedical Sciences: Faculty Publications

Objective: To evaluate the effect of serum antibody abundance against bovine coronavirus (BCV) on BCV shedding and risk of bovine respiratory disease (BRD) in beef calves from birth through the first 5 weeks in a feedlot.

Animals: 890 natural-service crossbred beef calves from 4 research herds.

Procedures: Serial blood samples for measurement of serum anti-BCV antibody abundance by an ELISA and nasal swab specimens for detection of BCV and other viral and bacterial BRD pathogens by real-time PCR methods were collected from all calves or subsets of calves at predetermined times from birth through the first 5 weeks after feedlot …

Trends In The Environmental Health Job Market For New Graduates, Jason W. Marion, Timothy J. Murphy Phd, Anne Marie Zimeri Phd

EKU Faculty and Staff Scholarship

The question of whether the job market can support future graduates of environmental health programs remains an important and difficult question for environmental health programs, current and prospective students, parents, and other stakeholders. Our previous report using 2014 data from the U.S. Bureau of Labor Statistics demonstrated anticipated growth and higher than average pay in the profession through at least 2022 for baccalaureate degree holders (Marion & Sinde, 2015). Growth in the profession does not necessarily translate into job availability if the market is saturated with job candidates. While university programs produce graduates, local health departments (LHDs) have suffered tremendous …

Cross-Reactive Immunity To Porcine Reproductive And Respiratory Syndrome Virus And Its Contribution To Protection, Ignacio Correas

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Due to the vast geographical distribution and significant economic losses generated, porcine reproductive and respiratory syndrome virus (PRRSV) can be considered the most important swine pathogen of contemporary times. Current control and eradication strategies against PRRSV have difficulty succeeding because of their complex nature and the absence of an effective vaccine. A major obstacle for PRRSV vaccine development is the broad heterogeneity of the virus, both at the genetic and antigenic level, its rapid evolution, and an incomplete knowledge of the immune responses responsible for clearing the virus from the host. Specifically, how known correlates of protection against PRRSV—neutralizing antibodies …

Hops (Humulus Lupulus L.) Bitter Acids: Modulation Of Rumen Fermentation And Potential As An Alternative Growth Promoter, Michael D. Flythe, Isabelle A. Kagan, Yuxi Wang, Nelmy Narvaez

Animal and Food Sciences Faculty Publications

Antibiotics can improve ruminant growth and efficiency by altering rumen fermentation via selective inhibition of microorganisms. However, antibiotic use is increasingly restricted due to concerns about the spread of antibiotic-resistance. Plant-based antimicrobials are alternatives to antibiotics in animal production. The hops plant (Humulus lupulus L.) produces a range of bioactive secondary metabolites, including antimicrobial prenylated phloroglucinols, which are commonly called alpha- and beta-acids. These latter compounds can be considered phyto-ionophores, phytochemicals with a similar antimicrobial mechanism of action to ionophore antibiotics (e.g., monensin, lasalocid). Like ionophores, the hop beta-acids inhibit rumen bacteria possessing a classical Gram-positive cell envelope. This …

A Parapoxviral Virion Protein Inhibits Nf-Κb Signaling Early In Infection, Sushil Khatiwada, Gustavo A. Delhon, Ponnuraj Nagendraprabhu, Sabal Chaulagain, Shuhong Luo, Diego G. Diel, Eduardo F. Flores, D. L. Rock

School of Veterinary and Biomedical Sciences: Faculty Publications

Poxviruses have evolved unique proteins and mechanisms to counteract the nuclear factor κB (NF-κB) signaling pathway, which is an essential regulatory pathway of host innate immune responses. Here, we describe a NF-κB inhibitory virion protein of orf virus (ORFV), ORFV073, which functions very early in infected cells. Infection with ORFV073 gene deletion virus (OV-IA82Δ073) led to increased accumulation of NF-κB essential modulator (NEMO), marked phosphorylation of IκB kinase (IKK) subunits IKKα and IKKβ, IκBα and NF-κB subunit p65 (NF-κB-p65), and to early nuclear translocation of NF-κB-p65 in virusinfected cells (30 min post infection). Expression of ORFV073 alone was sufficient to …

Developing A 670k Genotyping Array To Tag ~2m Snps Across 24 Horse Breeds, Robert J. Schaefer, Mikkel Schubert, Ernest F. Bailey, Danika L. Bannasch, Eric Barrey, Gila Kahila Bar-Gal, Gottfried Brem, Samantha A. Brooks, Ottmar Distl, Ruedi Fries, Carrie J. Finno, Vinzenz Gerber, Bianca Haase, Vidhya Jagannathan, Ted Kalbfleisch, Tosso Leeb, Gabriella Lindgren, Maria Susana Lopes, Núria Mach, Artur Da Câmara Machado, James N. Macleod, Annette Mccoy, Julia Metzger, Cecilia Penedo, Sagi Polani, Stefan Rieder, Imke Tammen, Jens Tetens, Georg Thaller, Andrea Verini-Supplizi

Gluck Equine Research Center Faculty Publications

Background: To date, genome-scale analyses in the domestic horse have been limited by suboptimal single nucleotide polymorphism (SNP) density and uneven genomic coverage of the current SNP genotyping arrays. The recent availability of whole genome sequences has created the opportunity to develop a next generation, high-density equine SNP array.

Results: Using whole genome sequence from 153 individuals representing 24 distinct breeds collated by the equine genomics community, we cataloged over 23 million de novo discovered genetic variants. Leveraging genotype data from individuals with both whole genome sequence, and genotypes from lower-density, legacy SNP arrays, a subset of ~5 million high-quality, …

Developing A Plant Virus-Based Expression System For The Expression Of Vaccines Against Porcine Reproductive And Respiratory Syndrome Virus, Hong Hanh Tran

Electronic Thesis and Dissertation Repository

Virus-based expression systems have been widely exploited for the production of recombinant proteins in plants during the last three decades. Advances in technology have boosted scale-up manufacturing of plant-made pharmaceuticals to high levels, via the complementation of transient expression and viral vectors. This combination allows proteins of interest to be produced in plants within a matter of days and thus, is well suited for the development of plant-made vaccines or therapeutics against emerging infectious diseases and potential bioterrorism agents. Several plant-based products are currently in varying stages of clinical development. To investigate the viability of virus-based expression systems for plant-made …

Volume 13, Number 1 (Spring/Summer 2017), Ut Institute Of Agriculture

Tennessee Land, Life and Science Magazine

Issue Highlights:

• The four pillars of the Institute of Agriculture
• Alumna forges partnership amid Waldo Canyon fire
• Fishery biologists revive a river

Growing South Dakota (Summer 2017), College Of Agriculture &. Biological Sciences

Growing South Dakota (Publication of the College of Agriculture, Food and Environmental Sciences)

This issue contains the 2016 SDSU Extension Annual Report.

Page] 2 From the Director
[Page] 3 WaterTouches Everything, Multi-State Network Strives to Enhance Water Stewardship
[Page] 6 From RubbleTo Renewal, Unique Art Project Helps Delmont Community Heal
[Page] 7 Future Focused, Community Development Program Helps Harness Ideas Into Action
[Page] 8 Health Help, Statewide Program Strives To Help Individuals Better Manage Chronic Health Condition
[Page] 10 Making An Impact, Master Gardener Program Helps Individuals & Communities Flourish
[Page] 12 Statewide Highlights & Happenings
[Page] 14 Outreach to Youth, Programs Foster Engagement With Diverse Youth Audiences
[Page] 17 A Look At …

Outcomes Of Spatially Fractionated Radiotherapy (Grid) For Bulky Soft Tissue Sarcomas In A Large Animal Model, Michael W. Nolan, Tracy L. Gieger, Alexander A. Karakashian, Mariana N. Nikolova‑Karakashian, Lysa P. Posner, Donald M. Roback, Judith N. Rivera, Sha Chang

Physiology Faculty Publications

GRID directs alternating regions of high- and low-dose radiation at tumors. A large animal model mimicking the geometries of human treatments is needed to complement existing rodent systems (eg, microbeam) and clarify the physical and biological attributes of GRID. A pilot study was undertaken in pet dogs with spontaneous soft tissue sarcomas to characterize responses to GRID. Subjects were treated with either 20 Gy (3 dogs) or 25 Gy (3 dogs), delivered using 6 MV X-rays and a commercial GRID collimator. Acute toxicity and tumor responses were assessed 2, 4, and 6 weeks later. Acute Radiation Therapy Oncology Group grade …

Investigation Of The Impact Of Increased Dietary Insoluble Fiber Through The Feeding Of Distillers Dried Grains With Solubles (Ddgs) On The Incidence And Severity Of Brachyspira-Associated Colitis In Pigs, Bailey Lauren Wilberts, Paulo Elias Arruda, Joann M. Kinyon, Timothy S. Frana, Chong Wang, Drew Robert Magstadt, Darin M. Madson, John F. Patience, Eric Ryan Burrough

Bailey (Wilberts) Arruda

Diet has been implicated as a major factor impacting clinical disease expression of swine dysentery and Brachyspira hyodysenteriae colonization. However, the impact of diet on novel pathogenic strongly beta-hemolytic Brachyspira spp. including “B. hampsonii” has yet to be investigated. In recent years, distillers dried grains with solubles (DDGS), a source of insoluble dietary fiber, has been increasingly included in diets of swine. A randomized complete block experiment was used to examine the effect of increased dietary fiber through the feeding of DDGS on the incidence of Brachyspira-associated colitis in pigs. One hundred 4-week-old pigs were divided …

Endocrine And Metabolic Effects Of Consuming Sugar-Sweetened Beverages: Preclinical And Clinical Studies", Peter Havel

Science Seminar Series

Peter J. Havel of the School of Veterinary Medicine at UC Davis will speak on his research on Endocrine and Metabolic Effects of Consuming Sugar-Sweetened Beverages: Preclinical and clinical studies at this Science Seminar Series lecture.

Central Role Of Vasotocin In The Neuroendocrine Regulation Of Stress Responses And Food Intake In Chickens, Gallus Gallus, Gurueswar Nagarajan

Graduate Theses and Dissertations

It is well known that arginine vasotocin (AVT) in birds is involved in physiological homeostasis such as cardiovascular, osmotic regulation as well as reproductive functions. Pertinent to these physiological functions, AVT immunoreactive (-ir) neurons in the hypothalamus have been found associated with hemorrhage, dehydration, oviposition and other physiological regulation. Evidence, however, suggests that AVT also plays significant roles in modulating behavior, memory, stress, and food intake. This dissertation research addresses the latter two neuroendocrine functions of AVT in detail within the chicken brain. First, the functional role of AVT-ir neurons in conjunction with corticotropin releasing hormone (CRH)-ir neurons in the …

Developing Learning Models To Teach Equine Anatomy And Biomechanics, Zandalee E. Toothaker

Honors College

Animal owners and professionals benefit from an understanding of an animal’s anatomy and biomechanics. This is especially true of the horse. A better understanding of the horse’s anatomy and weight bearing capabilities will allow people to treat and prevent injuries in equine athletes and work horses. Currently, teaching anatomy is based on dissection of cadavers and study of figures in textbooks. Dissection is expensive and presents logistical difficulties associated with sourcing, storage, and disposal of cadavers. Also, post mortem tissue changes obscure how the muscles support the bones and make the specimens unsuitable for representing biomechanical principles. Cadavers may also …

Georgia's Collaborative Approach To Expanding Mosquito Surveillance In Response To Zika Virus: A Case Study, R. Christopher Rustin, Deonte Martin, Varadan Sevilimedu, Sarbesh Pandeya, Haresh Rochani, Rosmarie Kelly

R. Christopher Rustin

Reduction In Activity By Noxious Chemical Stimulation Is Ameliorated By Immersion In Analgesic Drugs In Zebrafish, Javier Lopez-Luna, Qussay Al-Jubouri, Waleed Al-Nuaimy, Lynne U. Sneddon

Anesthesia and Analgesia Collection

Research has recently demonstrated that larval zebrafish show similar molecular responses to nociception to those of adults. Our study explored whether unprotected larval zebrafish exhibited altered behaviour after exposure to noxious chemicals and screened a range of analgesic drugs to determine their efficacy to reduce these responses. This approach aimed to validate larval zebrafish as a reliable replacement for adults as well as providing a high-throughput means of analysing behavioural responses. Zebrafish at 5 days postfertilization were exposed to known noxious stimuli: acetic acid (0.01%, 0.1% and 0.25%) and citric acid (0.1%, 1% and 5%). The behavioural response of each …

Growing South Dakota (Spring 2017), College Of Agriculture &. Biological Sciences

Growing South Dakota (Publication of the College of Agriculture, Food and Environmental Sciences)

[Page] 3 Opportunities With Oats SDSU Researchers Strive to Improve Varieties [Page] 6 Stopping Stem Canker Plant Pathologists Making Progress To Keep This Disease At Bay
[Page] 7 Disease Detectives Vet Scientists Researching Another Emerging Virus [Page] 8 Community Support Devastating West River Fire Brings Ag Industry Together
[Page] 10 Research In Progress Dakota Lakes Research Farm Investigates Three Big Ideas
[Page] 14 Water Quality Quest Understanding E. Coli Behavior Aims To Aid Water Quality In Rivers & Streams
[Page] 15 Verifying Ag Land Valuation Researchers Assist In Reviewing Ag Land’s Classification
[Page] 16 Cattle Insights Fetal Programming Research Evaluates …