Management of chronic wasting disease in ranched elk: conclusions from a longitudinal three-year study.

Chronic wasting disease is a fatal, horizontally transmissible prion disease of cervid species that has been reported in free-ranging and farmed animals in North America, Scandinavia, and Korea. Like other prion diseases, CWD susceptibility is partly dependent on the sequence of the prion protein encoded by the host's PRNP gene; it is unknown if variations in PRNP have any meaningful effects on other aspects of health. Conventional diagnosis of CWD relies on ELISA or IHC testing of samples collected post-mortem, with recent efforts focused on antemortem testing approaches. We report on the conclusions of a study evaluating the role of antemortem testing of rectal biopsies collected from over 570 elk in a privately managed herd, and the results of both an amplification assay (RT-QuIC) and conventional IHC among animals with a several PRNP genotypes. Links between PRNP genotype and potential markers of evolutionary fitness, including pregnancy rates, body condition, and annual return rates were also examined. We found that the RT-QuIC assay identified significantly more CWD positive animals than conventional IHC across the course of the study, and was less affected by factors known to influence IHC sensitivity - including follicle count and PRNP genotype. We also found that several evolutionary markers of fitness were not adversely correlated with specific PRNP genotypes. While the financial burden of the disease in this herd was ultimately unsustainable for the herd owners, our scientific findings and the hurdles encountered will assist future CWD management strategies in both wild and farmed elk and deer.

Cross-validation of the RT-QuIC assay for the antemortem detection of chronic wasting disease in elk.

Chronic wasting disease is a progressively fatal, horizontally transmissible prion disease affecting several members of the cervid species. Conventional diagnosis relies on ELISA or IHC evaluation using tissues collected post-mortem; however, recent research has focused on newly developed amplification techniques using samples collected antemortem. The present study sought to cross-validate the real-time quaking-induced conversion assay (RT-QuIC) evaluation of rectal biopsies collected from an elk herd with endemic CWD, assessing both binary positive/negative test results as well as relative rates of amplification between laboratories. We found that results were correlative in both categories across all laboratories performing RT-QuIC, as well as to conventional IHC performed at a national reference laboratory. A significantly higher number of positive samples were identified using RT-QuIC, with results seemingly unhindered by low follicle counts. These findings support the continued development and implementation of amplification assays in the diagnosis of prion diseases of veterinary importance, targeting not just antemortem sampling strategies, but post-mortem testing approaches as well.

T1ρ MRI of the talar articular cartilage is increased in those with chronic ankle instability.

OBJECTIVE: To determine if individuals with chronic ankle instability (CAI) demonstrate different talar cartilage T1ρ relaxation times compared to uninjured controls. DESIGN: Fifteen CAI (21.13 ± 1.81 years, 4.00 ± 2.07 previous ankle sprains) and fifteen controls (21.07 ± 2.55 years, no previous ankle sprains) participated. CAI inclusion criteria was in accordance with the International Ankle Consortium guidelines. Greater T1ρ relaxation times were interpreted as greater degenerative changes. Participants were non-weight bearing for 30-minutes prior to scanning to unload the cartilage. Voxel by voxel T1ρ relaxation times were calculated from a five image sequence. Segmentation of the talar cartilage was performed manually using ITK-SNAP software. T1ρ relaxation time means and variability across the entire talus and in the anteromeidal, anterolateral, posteromedial, and posterolateral regions of interest (ROIs) were compared between groups using mean differences and effect sizes (ES) with their corresponding 95% confidence intervals (95%CI). RESULTS: Individuals with CAI demonstrated higher T1ρ relaxation times (mean ± standard deviation) across the entire talus (CAI: 65.97 ± 10.45 ms, Control: 58.84 ± 7.68 ms; ES = 0.76, 95%CI = 0.02-1.50), in the anterolateral (ES = 1.00, 95%CI = 0.24-1.48), posteromedial (ES = 0.74, 95%CI = 0.01-1.49), and posterolateral region of interest (ES = 3.84, 95%CI = 2.63-5.04). The T1ρ relaxation time variability (mean ± standard deviation) also differed across the overall talus (CAI: 32.78 ± 4.06 ms, Control: 28.23 ± 4.45 ms; ES = 1.04, 95%CI = 0.28-1.80), in the anteriolateral, (ES = 1.07, 95%CI = 0.31, 1.84) and posteriolateral (ES = 1.00, 95%CI = 0.24-1.75) ROIs. CONCLUSIONS: Individuals with CAI demonstrate greater T1ρ relaxation times and higher T1ρ variability compared to uninjured controls. This finding supports the existing literature illustrating early degenerative joint tissue changes consistent with early onset posttraumatic osteoarthritis in individuals with CAI.

PO and ID BCG vaccination in humans induce distinct mucosal and systemic immune responses and CD4+ T cell transcriptomal molecular signatures.

Protective efficacy of Bacillus Calmette-Guérin (BCG) may be affected by the methods and routes of vaccine administration. We have studied the safety and immunogenicity of oral (PO) and/or intradermal (ID) administration of BCG in healthy human subjects. No major safety concerns were detected in the 68 healthy adults vaccinated with PO and/or ID BCG. Although both PO and ID BCG could induce systemic Th1 responses capable of IFN-γ production, ID BCG more strongly induced systemic Th1 responses. In contrast, stronger mucosal responses (TB-specific secretory IgA and bronchoalveolar lavage T cells) were induced by PO BCG vaccination. To generate preliminary data comparing the early gene signatures induced by mucosal and systemic BCG vaccination, CD4+ memory T cells were isolated from subsets of BCG vaccinated subjects pre- (Day 0) and post-vaccination (Days 7 and 56), rested or stimulated with BCG infected dendritic cells, and then studied by Illumina BeadArray transcriptomal analysis. Notably, distinct gene expression profiles were identified both on Day 7 and Day 56 comparing the PO and ID BCG vaccinated groups by GSEA analysis. Future correlation analyses between specific gene expression patterns and distinct mucosal and systemic immune responses induced will be highly informative for TB vaccine development.

Zinc regulates the function and expression of the iron transporters DMT1 and IREG1 in human intestinal Caco-2 cells.

Trace metals influence the absorption of each other from the diet and it has been suggested that the divalent metal transporter (DMT1) represents a common uptake pathway for these important micronutrients. However, compelling evidence from our laboratory suggests that DMT1 is predominantly an iron transporter, with lower affinity for other metals. Several studies have shown that increasing dietary iron downregulates DMT1. Interestingly, our current data indicate that zinc upregulates DMT1 protein and mRNA expression and also pH-dependent iron uptake. Transepithelial flux of iron was also increased and was associated with a rise in IREG1 mRNA expression.

Breakdown of extracellular ATP by the prostatic and epididymal ends of the guinea pig vas deferens.

Adenosine 5'-triphosphate (ATP) has a higher potency at the prostatic than at the epididymal end of the guinea pig vas deferens. We bisected the tissue and measured the breakdown of ATP by each half, and although the half-lives differed, the rate constants per gram of tissue were not significantly different. For a range of tissue portions, a correlation was found between the portion weight and the half-life of ATP. The difference in half-life at the two ends is therefore due to the different weights of the tissues, and the difference in potency of ATP cannot be explained by differences in degradation.

Effects of allopurinol, erythro-9-(2-hydroxy-3-nonyl)adenine and S-(4-nitrobenzyl)-6-thioinosine on the degradation of adenosine 5'-triphosphate in the rat colon muscularis mucosae.

The effects on ATP breakdown of some modulators of adenosine transport or metabolism were studied in the rat colon muscularis mucosae, a tissue which contracts to ATP and is thought to contain P2Y1 receptors. The compounds tested were the xanthine oxidase inhibitor allopurinol, the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and the adenosine uptake blocker S-(4-nitrobenzyl)-6-thioinosine (NBTI). The degradation of adenosine 5'-triphosphate (ATP) (100 microM) and the appearance of metabolites was followed by high pressure liquid chromatography during incubation of isolated tissue preparations alone or in the presence of the drugs, following preincubation with the drugs for 1 h. In the absence of drugs ATP was rapidly degraded by the rat colon muscularis mucosae with a half-life of 6.1 +/- 0.7 min, the major breakdown product being inosine rather than adenosine. Allopurinol (1 microM) and NBTI (10 microM) had no effect on the rate of breakdown of ATP or on the pattern of metabolites produced. EHNA (1 or 10 microM) also had no effect on the half-life of ATP, but in the presence of EHNA (1 microM) the rate of production of inosine was significantly reduced and some adenosine was detected, while in the presence of 10 microm EHNA the production of inosine was abolished and adenosine became the final breakdown product. These results indicate that allopurinol (1 microM) and NBTI (10 microM) have no detectable effect on extracellular purine metabolism in this tissue, and that the build-up of adenosine produced by treatment with EHNA does not have a feedback effect on ATP breakdown.

Effects of noradrenaline, the calcium ionophore A23187, forskolin, sodium nitroprusside and glibenclamide on the degradation of extracellular adenosine 5'-triphosphate by the rat isolated vas deferens.

1. The effects of noradrenaline (NA), the calcium ionophore A23187, forskolin, sodium nitroprusside (SNP) and the K+-channel blocker glibenclamide on the degradation by ectonucleotidases of extracellular adenosine 5'-triphosphate (ATP) were studied in the rat vas deferens. 2. ATP (100 microM) was rapidly broken down by the rat vas deferens with a half-life of 5.83 +/- 0.40 min via adenosine 5'-diphosphate (ADP) and adenosine 5'-monophosphate (AMP), with the final degradation product being inosine and with little adenosine being detected in the samples. 3. Preincubation for 1 h with NA (10 microM), A23187 (10 microM), or glibenclamide (100 microM) had no significant effect on the breakdown of ATP or the production of metabolites. However, both forskolin (10 microM) and SNP (1 microM) significantly increased the concentrations of AMP detected with time. In the case of SNP (1 microM) there was also a significant reduction in the rate of production of inosine, while in the case of forskolin (10 microM) there was a significant increase in the rate of removal of ATP. 4. These results suggest that preincubation with SNP may inhibit 5'-nucleotidase and so reduce the metabolism of AMP, while preincubation with forskolin may increase the activity of the ectonucleotidases responsible for production of AMP from ATP.

Persistence and boosting of bacille Calmette-Guérin-induced delayed-type hypersensitivity.

BACKGROUND: Bacille Calmette-Guérin (BCG) vaccination may induce persistent and booster purified protein derivative (PPD) responses that complicate tuberculosis screening efforts. OBJECTIVES: To investigate the effects of BCG vaccination on serial PPD tests and to study correlations between persistent delayed-type hypersensitivity and other potential surrogate markers of protective immunity. DESIGN: Cohort study. SETTING: Midwestern urban university. PARTICIPANTS: 69 healthy adults. INTERVENTIONS: BCG vaccination, blood samples drawn for immunologic studies, and PPD tests done sequentially over 1 to 3 years. MEASUREMENTS: Serial PPD induration, lymphoproliferation, and interferon-gamma responses. RESULTS: 10% of participants (95% CI, 4% to 20%) had persistent PPD responses of 15 mm or greater, and 3% (CI, 0% to 10%) demonstrated PPD boosting of 15 mm or greater 1 to 3 years after BCG vaccination. Intradermal BCG vaccination induced a larger number of persistent responses that were 10 mm or greater than did percutaneous BCG vaccination (12 of 46 participants compared with 1 of 23 participants; P = 0.05). Persistent and boosted delayed-type hypersensitivity correlated with mycobacterial-specific lymphoproliferation and interferon-gamma responses. CONCLUSIONS: Previous BCG vaccination reduces the predictive value of serial PPD testing. The lowest PPD predictive values will occur in persons without known tuberculosis exposure who were vaccinated recently or many times with intradermal BCG. In addition, BCG-related persistence and boosting of delayed-type hypersensitivity responses correlate with other potential surrogate markers of protective mycobacterial immunity.

Clinical reactogenicity of intradermal bacille Calmette-Guérin vaccination.

Clinical, microbiological, and immunologic responses were evaluated in volunteers vaccinated intradermally with bacille Calmette-Guérin (BCG). Most volunteers (98%) developed ulcerative lesions that drained for a mean +/- SE of 4.3 +/- 0.29 weeks. Mycobacterial DNA was detected by a polymerase chain reaction-based amplification technique in biopsy specimens from BCG ulcers 2 weeks after vaccination and in blood specimens 3 days after vaccination. Mycobacteria were cultured from ulcer drainage 2 months after vaccination, demonstrating a prolonged potential risk of contact spread of the vaccine strain. The duration of ulcer drainage was inversely correlated with prevaccination lymphoproliferative (r = -0.515; P < .002) and interferon gamma (r = -0.841; P < .002) responses specific to mycobacteria and directly correlated with postvaccination increases in lymphoproliferative (r = 0.498; P < .002) and interferon gamma (r = 0.688; P < .02) responses specific to mycobacteria. These results demonstrate the clinical reactogenicity of BCG and the potential risk of contact spread of the vaccine strain and suggest that clinical reactogenicity is a trade-off for the induction of protective mycobacterial immunity.

Effects of adenosine 5'-triphosphate, uridine 5'-triphosphate, adenosine 5'-tetraphosphate and diadenosine polyphosphates in guinea-pig taenia caeci and rat colon muscularis mucosae.

The functional effects of adenosine 5'-triphosphate (ATP), uridine 5'-triphosphate (UTP), adenosine 5'-tetraphosphate (AP4) and the diadenosine polyphosphates P1,P3-diadenosine triphosphate (Ap3A), P1,P4-diadenosine tetraphosphate (Ap4A) and P1,P5-diadenosine pentaphosphate (Ap5A) were studied in two isolated smooth muscle preparations thought to contain P2Y (P2Y1) receptors, the guinea-pig taenia caeci (which relaxes to ATP) and the rat colon muscularis mucosae (which contracts to ATP). In addition, the breakdown of these compounds by the rat colon muscularis mucosae was investigated by high pressure liquid chromatography. In the guinea-pig taenia caeci all the purine nucleotides caused relaxation with a potency order of Ap3A=Ap4A> ATP>AP4=Ap5A, and these relaxations were antagonised by suramin with apparent pA2 values in the region of 5, consistent with activation of a P2Y1 receptor. In the rat colon muscularis mucosae the nucleotides caused contraction with a potency order of Ap3A = Ap4A>ATP=AP4 =Ap5A >UTP. However, while suramin (100 microM) inhibited responses to ATP and UTP at all concentrations of agonist, it only inhibited contractions induced by the higher concentrations of AP4, Ap3A and Ap4A and had little effect on contractions induced by Ap5A. A higher concentration of suramin (1 mM) enhanced contractions induced by ATP but greatly inhibited those induced by UTP and had no effect on responses to the other agonists. The A1 adenosine receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX; 10 nM) had no effect on responses to ATP or UTP but inhibited responses to Ap3A, Ap4A, Ap5A and AP4. A combination of suramin (1 mM) and DPCPX (10 nM) almost abolished responses to all the agonists. ATP and UTP were rapidly degraded by the rat colon muscularis mucosae while AP4, Ap3A, Ap4A and Ap5A were degraded more slowly, and the major product detected after breakdown of the purine nucleotides was inosine rather than adenosine. The breakdown of all the nucleotides was inhibited by suramin (1 mM), although this inhibition did not achieve statistical significance in the case of ATP. These results show that while the diadenosine polyphosphates appear to act as P2 agonists in the taenia caeci, in the rat colon muscularis mucosae their major action is via adenosine A1 receptors rather than via P2 receptors. In addition, although they are more stable than ATP or UTP, their action in this tissue is clearly affected by their degradation which complicates the effects of suramin.

Participant assessment of a nonmedical breast cancer support group.

BACKGROUND: This study was designed to examine how a particular format of a support group for women with breast cancer was related to the use of the group and experiences of the participants. SUBJECTS: A total of 131 women who participated in a breast cancer support group under the auspices of the YWCA were interviewed. RESULTS: Findings replicated existing studies indicating such generic benefits of support group participation as enhanced sense of control, sharing of information and experiences, and acquaintance with positive role models. In addition to these benefits, participants reacted favorably to the opportunities provided by a drop-in format, an exercise component, and opportunities for sharing in a nonmedical setting. Active use of the group was associated with lack of emotional support from family. Active use also was associated with favorable reported outcomes for well-being and health.

An innovative night service program in home care.

Comprehensive night service by a home care agency has resulted in increased customer satisfaction and referrals to the agency. The description of this innovative night nurse program includes the job requirements and duties of a visiting night nurse and the tools the nurse uses to accomplish the task of nighttime care delivery. The use of self-directed work groups and mentorship to manage the demands of night service are discussed. Continuity of care, nurse safety, professional development, and program cost also are addressed.

Environmental assessment.

The significance, standard elements, components, and documentation of an environmental assessment in home care are discussed. This assessment is delineated within Maslow's Hierarchy of Needs and from a functional perspective. An Environmental Assessment Form that can be used as a documentation tool is included.

Achieving success in home care through the self-directed work group approach.

Self-directed work groups (SDWGs) have become the hallmark of many successful corporations. This work-management philosophy is a cost-effective way to respond quickly to the challenges of today's rapidly changing home healthcare environment. To enable nurses to develop SDWGs, the five key characteristics of these groups--coaching, communication, commitment, cooperation, and contribution--are described.

Forward light scatter at one month after photorefractive keratectomy.

BACKGROUND: Although it is known that backward light scatter increases transiently following most excimer laser photorefractive keratectomies (PRKs), it is not clear that there is a significant increase in forward light scatter, which is of primary concern for the patient. The object of this study was to determine if there is a significant change of forward light scatter at 1 month after (PRK) with an ablation zone diameter of 6 mm. METHODS: Overlapping subsets of 24 normal myopic eyes were tested before (on the day of surgery) and 1 month after PRK, using three instruments: a Stray Light Meter (16 eyes); a Computerized Stray Light Meter (14 eyes); and a mesopic Increment Threshold-Glare Paradigm (six eyes). Differences between the two eyes before PRK were compared with the differences between the same eye before and after PRK, using repeated measured analysis of variance. In addition, increment threshold data obtained from 22 eyes after PRK were compared with those of 60 controls of the same age range and distribution by a t test. RESULTS: None of the statistical comparisons approached significance at the alpha = 0.05 level. Changes in light scatter as small as a factor of 1.95 (Stray Light Meter) and 1.55 (Increment Threshold-Glare Paradigm) could be detected as significant with a high power (0.8). Changes larger than a factor of 21 could be detected with a power of 0.8 for the Computerized Stray Light Meter. CONCLUSIONS: In these data, there is no support for the hypothesis that forward light scatter increases significantly 1 month after PRK with an ablation zone of 6 mm. Any increases in forward light scatter are unlikely to be greater than a factor of 1.5 to 2 under daytime or nighttime illumination conditions.