Detection of prions from spiked and free-ranging carnivore feces.

Chronic wasting disease (CWD) is a highly contagious, fatal neurodegenerative disease caused by infectious prions (PrPCWD) affecting wild and captive cervids. Although experimental feeding studies have demonstrated prions in feces of crows (Corvus brachyrhynchos), coyotes (Canis latrans), and cougars (Puma concolor), the role of scavengers and predators in CWD epidemiology remains poorly understood. Here we applied the real-time quaking-induced conversion (RT-QuIC) assay to detect PrPCWD in feces from cervid consumers, to advance surveillance approaches, which could be used to improve disease research and adaptive management of CWD. We assessed recovery and detection of PrPCWD by experimental spiking of PrPCWD into carnivore feces from 9 species sourced from CWD-free populations or captive facilities. We then applied this technique to detect PrPCWD from feces of predators and scavengers in free-ranging populations. Our results demonstrate that spiked PrPCWD is detectable from feces of free-ranging mammalian and avian carnivores using RT-QuIC. Results show that PrPCWD acquired in natural settings is detectable in feces from free-ranging carnivores, and that PrPCWD rates of detection in carnivore feces reflect relative prevalence estimates observed in the corresponding cervid populations. This study adapts an important diagnostic tool for CWD, allowing investigation of the epidemiology of CWD at the community-level.

Ticks harbor and excrete chronic wasting disease prions.

Chronic wasting disease (CWD) is a fatal neurodegenerative disease caused by infectious prions (PrPCWD) affecting cervids. Circulating PrPCWD in blood may pose a risk for indirect transmission by way of hematophagous ectoparasites acting as mechanical vectors. Cervids can carry high tick infestations and exhibit allogrooming, a common tick defense strategy between conspecifics. Ingestion of ticks during allogrooming may expose naïve animals to CWD, if ticks harbor PrPCWD. This study investigates whether ticks can harbor transmission-relevant quantities of PrPCWD by combining experimental tick feeding trials and evaluation of ticks from free-ranging white-tailed deer (Odocoileus virginianus). Using the real-time quaking-induced conversion (RT-QuIC) assay, we show that black-legged ticks (Ixodes scapularis) fed PrPCWD-spiked blood using artificial membranes ingest and excrete PrPCWD. Combining results of RT-QuIC and protein misfolding cyclic amplification, we detected seeding activity from 6 of 15 (40%) pooled tick samples collected from wild CWD-infected white-tailed deer. Seeding activities in ticks were analogous to 10-1000 ng of CWD-positive retropharyngeal lymph node collected from deer upon which they were feeding. Estimates revealed a median infectious dose range of 0.3-42.4 per tick, suggesting that ticks can take up transmission-relevant amounts of PrPCWD and may pose a CWD risk to cervids.

Economics of timed artificial insemination with unsorted or sexed semen in a high-producing, pasture-based dairy production system.

This study used a stochastic simulation model to estimate the potential economic benefit of using timed artificial insemination (TAI) in combination with conventional unsorted (TCONV) and sexed (TSEX) semen in heifers only (TCONV-H, TSEX-H) and in both heifers and lactating cows (TCONV-HC, TSEX-HC) in a high-producing, pasture-based production system. The scenarios were compared with a conventional reproductive policy (CONV) in which heifers and cows were inseminated with conventional unsorted semen after estrus detection. Sensitivity analysis was also used to estimate the effect of hormone costs from TAI use on the profitability of each program relative to CONV. The mean annual (± standard deviation) profit advantage (ΔPROF) over CONV for TCONV-H, TCONV-HC, TSEX-H, and TSEX-HC scenarios were €3.90/cow ± 4.65, €34.11/cow ± 25.69, €13.96/cow ± 6.83, and €41.52/cow ± 42.86, respectively. Combined application of both technologies was shown to return a greater annual ΔPROF on average compared with that achievable from TAI alone. However, the risk of not returning a positive annual ΔPROF varied across the scenarios with higher risk in TCONV-H and TSEX-HC. Specifically, TCONV-H and TSEX-HC had a 24 and 18% chance, respectively, of not returning a positive annual ΔPROF. Sensitivity analysis showed that when hormone costs increased by €10/cow TCONV-H and TSEX-HC had a 38 and 23% chance, respectively, of not returning a positive annual ΔPROF. The range in ΔPROF for TCONV policies was most sensitive to the TAI pregnancy rate and TSEX policies were most sensitive to the relative fertility achieved with sexed compared with unsorted semen. This study has shown TAI and sexed semen are complementary technologies that can increase genetic gain and profitability in a pasture-based, dairy production system.

Effects of herd fertility on the economics of sexed semen in a high-producing, pasture-based dairy production system.

This study used a stochastic simulation model to estimate the potential economic benefit of using sexed semen in heifers only and in heifers and lactating cows in a high-producing, pasture-based system under 3 fertility scenarios. Three breeding strategies were modeled: (1) only heifers inseminated with sexed semen and cows inseminated with conventional unsexed semen (SSH); (2) both heifers and cows inseminated with sexed semen (SSHC); and (3) a reference scenario in which all females were inseminated with conventional, unsexed semen (CONV). Each scenario was evaluated under 3 herd fertility states: high (HF), medium (MF), and low (LF), which, under the reference scenario, corresponded to herd replacement rates of 21, 25, and 31%, respectively. The model estimated the economic profit, including the net present value of the genetic gain from selection intensity. The economic return from adoption of sexed semen strategies declined, with reduced levels of baseline herd fertility turning negative in the LF state. The mean (±SD) sexed semen advantage (SSA) per cow for HF-SSH, MF-SSH, and LF-SSH scenarios were €30.61 ± 8.98, €27.45 ± 7.19, and €14.69 ± 11.06, respectively. However, the SSA per cow for HF-SSHC, MF-SSHC, and LF-SSHC scenarios were €49.14 ± 15.43, €18.46 ± 30.08, and -€19.30 ± 57.11. The range in economic profit for SSA for SSH was most sensitive to calf prices in HF-SSH and the pregnancy rate of sexed semen as a percentage of conventional unsorted semen in MF-SSH and LF-SSH. The range in economic profit for SSA for SSHC scenarios was most sensitive to the pregnancy rate of sexed semen as a percentage of conventional unsorted semen in HF-SSHC, MF-SSHC, and LF-SSHC. This study highlights the effect of baseline herd fertility state on the financial advantage of adopting sexed semen in a pasture-based dairy production system.

An efficient, non-viral dendritic vector for gene delivery in tissue engineering.

Recent developments within the field of tissue engineering (TE) have shown that biomaterial scaffold systems can be augmented via the incorporation of gene therapeutics. The objective of this study was to assess the potential of the activated polyamidoamine dendrimer (dPAMAM) transfection reagent (SuperfectTM) as a gene delivery system to mesenchymal stem cells (MSCs) in both monolayer and 3D culture on collagen based scaffolds. dPAMAM-pDNA polyplexes at a mass ratio (M:R) 10:1 (dPAMAM : pDNA) (1 ug pDNA) were capable of facilitating prolonged reporter gene expression in monolayer MSCs which was superior to that facilitated using polyethylenimine (PEI)-pDNA polyplexes (2 ug pDNA). When dPAMAM-pDNA polyplexes (1 ug pDNA) were soak loaded onto a collagen-chondroitin sulphate (CS) scaffold prolonged transgene expression was facilitated which was higher than that obtained for a PEI-pDNA polyplex (2 ug pDNA) loaded scaffold. Transgene expression was dependent on the composite nature of the collagen scaffold with varying expression profiles obtained from a suite of collagen constructs including a collagen alone, collagen-CS, collagen-hydroxyapatite, collagen-nanohydroxyapatite and collagen-hyaluronic acid scaffold. Therefore, the dPAMAM vector described herein represents a biocompatible, effective gene delivery vector for TE applications which, via matching with a particular composite scaffold type, can be tailored for regeneration of various tissue defects.

Recent advances in small molecule microarrays: applications and technology.

The field of Small Molecule Microarray's (SMM's) is an ever-expanding part of the larger microarray field. SMM's are array based detection systems that use small molecules as probes immobilized on a variety of microarray surfaces that are screened against a number of targets for purposes including, but not limited to, protein-small molecule ligand recognition and protein function profiling. This review covers the recent advances in the field with particular emphasis on the successful applications of SMM's, as well as technical advances in platform optimization and novel small molecule immobilization strategies.

Systematic Study of Chromatographic Behavior vs Alkyl Chain Length for HPLC Bonded Phases Containing an Embedded Carbamate Group.

A series of HPLC bonded phases containing an internal carbamate group were studied by changing the terminal N-alkyl group from C(8)H(17) to C(18)H(37) in increments of two methylene units, i.e., C(8), C(10), C(12), C(14), C(16), and C(18). Each material was prepared via bonding of silica with the respective 3-(chlorodimethylsilyl)propyl N-alkylcarbamate silane. The materials were compared under reversed-phase conditions using a test mixture of nonpolar, polar, and basic compounds in a 65:35 (v/v) methanol/20 mM KH(2)PO(4)/K(2)HPO(4), pH 7, mobile phase. Retention factors were found to generally increase from the C(8) length to the C(12)-C(16) lengths but decreased for the C(18) length. Retention factors were also measured as a function of three ligand surface concentrations for the C(12), C(14), and C(18) materials. In this study, retention generally decreased with increasing surface concentration, especially for the C(18) chain length. Changes in particle surface area and porosity caused by bonding did not fully account for the observed changes in retention factors. Peak shapes for the basic analytes propranolol and amitriptyline were also studied as a function of N-alkylcarbamate chain length and surface concentration. Tailing factors were unaffected by chain length and only weakly dependent on surface concentration. By comparison, tailing factors decreased significantly as surface concentration increased for a set of conventional C(18) alkyl packings.

The effectiveness of the advanced trauma life support system in a mass casualty situation by non-trauma-experienced physicians: Grenada 1983.

Mass casualty is a sporadic event precipitated by natural or man made causes which can be defined as the need for medical care exceeding the ability to provide it. Many literature reports of mass casualty evolutions depict scenes of chaos and confusion, leading to a need for a standardized approach to assessment, triage, and initial resuscitation. Even with the advent of trained emergency medicine specialists to direct these activities, such a framework would seem highly desirable for other participating primary care specialists. Additionally, a uniform system might be particularly useful in the mass casualty situation where international rescue teams converge on one disaster site. Advanced Trauma Life Support (ATLS) is a standardized approach easily adaptable to triage and resuscitation of multiple patients. Its use and effectiveness in mass casualty, however, has not had prior mention in the literature. This paper presents the first reported adaptation of ATLS principles to mass casualty during the invasion of Grenada. The bulk of 76 patients brought to the Primary Casualty Receiving and Treatment Center (PCRTC) were triaged, stabilized, and resuscitated by three PGY-1 trained, non-trauma-experienced physicians. During the primary survey, 8 major life threatening problems were identified and immediately corrected without loss of life. The ATLS system seemed to provide a comfortable framework for these partially trained physicians. Arguments for its adaptation and use as an international system approach are discussed.