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Objective: Behavioral parent training has been proven effective through years of research with a variety of groups. However, little research has been conducted to systematically evaluate the extent to which behavioral parent training may improve parenting skills of foster and other caregivers of dependent children. The Behavior Analysis Services Program (BASP) has been charged with providing behavioral parent training to foster parents and other caregivers of dependent children throughout the state of Florida through a variety of services focusing primarily on a group-class-based training curriculum. This article examines the extent to which behavioral parent training was effective in increasing skills of caregivers who were trained in 2001 and 2002. Methods: Study 1 evaluated skills acquisition via a large-scale analysis of precourse and postcourse assessments, as well as observations of in-home parent-child interactions. Study 2 assessed skills acquisition via controlled analyses using repeated measures and multiple baseline designs. Results: Results showed increases in parenting skills in both studies as a function of curriculum training. Discussion: Implications of the results, limitations, and directions for future research are discussed. [ABSTRACT FROM AUTHOR]

Background: The w Mel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and subsequently shown to reduce transmission of dengue and other pathogens, under both laboratory and field conditions. Here we describe the entomological outcomes of w Mel Wolbachia mosquito releases in two small communities in Nha Trang City in central Vietnam. Methods: The w Mel strain of Wolbachia was backcrossed into local Aedes aegypti genotype and mosquito releases were undertaken by community members or by staff. Field monitoring was undertaken to track Wolbachia establishment in local Ae. aegypti mosquito populations. Ecological studies were undertaken to assess relationships between environmental factors and the spatial and temporal variability in Wolbachia infection prevalence in mosquitoes. Results: Releases of w Mel Wolbachia Ae. aegypti mosquitoes in two small communities in Nha Trang City resulted in the initial establishment of Wolbachia in the local Ae. aegypti mosquito populations, followed by seasonal fluctuations in Wolbachia prevalence. There was significant small-scale spatial heterogeneity in Wolbachia infection prevalence in the Tri Nguyen Village site, resulting in the loss of w Mel Wolbachia infection in mosquitoes in north and center areas, despite Wolbachia prevalence remaining high in mosquitoes in the south area. In the second site, Vinh Luong Ward, Wolbachia has persisted at a high level in mosquitoes throughout this site despite similar seasonal fluctuations in w Mel Wolbachia prevalence. Conclusion: Seasonal variation in Wolbachia infection prevalence in mosquitoes was associated with elevated temperature conditions, and was possibly due to imperfect maternal transmission of Wolbachia . Heterogeneity in Wolbachia infection prevalence was found throughout one site, and indicates additional factors may influence Wolbachia establishment., Competing Interests: No competing interests were disclosed., (Copyright: © 2022 Hien NT et al.)

Objectives: Host gene expression signatures discriminate bacterial and viral infection but have not been translated to a clinical test platform. This study enrolled an independent cohort of patients to describe and validate a first-in-class host response bacterial/viral test., Design: Subjects were recruited from 2006 to 2016. Enrollment blood samples were collected in an RNA preservative and banked for later testing. The reference standard was an expert panel clinical adjudication, which was blinded to gene expression and procalcitonin results., Setting: Four U.S. emergency departments., Patients: Six-hundred twenty-three subjects with acute respiratory illness or suspected sepsis., Interventions: Forty-five-transcript signature measured on the BioFire FilmArray System (BioFire Diagnostics, Salt Lake City, UT) in ~45 minutes., Measurements and Main Results: Host response bacterial/viral test performance characteristics were evaluated in 623 participants (mean age 46 yr; 45% male) with bacterial infection, viral infection, coinfection, or noninfectious illness. Performance of the host response bacterial/viral test was compared with procalcitonin. The test provided independent probabilities of bacterial and viral infection in ~45 minutes. In the 213-subject training cohort, the host response bacterial/viral test had an area under the curve for bacterial infection of 0.90 (95% CI, 0.84-0.94) and 0.92 (95% CI, 0.87-0.95) for viral infection. Independent validation in 209 subjects revealed similar performance with an area under the curve of 0.85 (95% CI, 0.78-0.90) for bacterial infection and 0.91 (95% CI, 0.85-0.94) for viral infection. The test had 80.1% (95% CI, 73.7-85.4%) average weighted accuracy for bacterial infection and 86.8% (95% CI, 81.8-90.8%) for viral infection in this validation cohort. This was significantly better than 68.7% (95% CI, 62.4-75.4%) observed for procalcitonin (p < 0.001). An additional cohort of 201 subjects with indeterminate phenotypes (coinfection or microbiology-negative infections) revealed similar performance., Conclusions: The host response bacterial/viral measured using the BioFire System rapidly and accurately discriminated bacterial and viral infection better than procalcitonin, which can help support more appropriate antibiotic use., Competing Interests: Dr. Tsalik received in-kind support from BioFire Diagnostics by way of consumables and test instruments; received funding from Predigen, Inc.. BioFire, Inc. provided in-kind support for test development reagents used in this study. Drs. Tsalik, Henao, McClain, Ginsburg, Burke, and Woods disclosed filing for a patent pertaining to the signatures discussed in this study (WO 2017/004390 A1). Dr. Montgomery was an employee of BioFire Diagnostics, LLC. Dr. Nawrocki disclosed he has shares in BioMérieux. Dr. Lydon was supported by the Eugene A. Stead Scholarship from Duke University School of Medicine and the Infectious Diseases Society of America Medical Scholars Program. Drs. Tsalik, Ginsburg, and Woods disclosed that they are cofounders of Predigen, Inc. Drs. Tsalik, Ko, Petzold, Cairns, Kingsmore, Fowler, Ginsburg, Burke, and Woods received support for article research from the National Institutes of Health (NIH). Drs. Nawrocki and Hemmert received funding from BioFire Diagnostics, LLC.; and disclosed that they are employees of BioFire Diagnostics, LLC. Dr. Cairns is a consultant for BioMérieux, Inc. Dr. Ko’s institution received funding from the Antibiotic Resistance Leadership Group; disclosed the off-label product use of diagnostic tests. Dr. Petzold received support for article research from the Defense Advanced Research Projects Agency (DARPA) (NIH National Institute of Allergy and Infectious Diseases (NIAID) U01AI066569 and UM1AI104681 U.S. DARPA contract—N66001-09-C2082). Dr. Cairns’ institution received funding from the NIH (NIAID) and the DARPA; and received funding from BioMérieux. Dr. Kingsmore’s institution received funding from the NIH. Dr. Fowler received funding from the NIH, MedImmune, Allergan, Pfizer, Advanced Liquid Logics, Theravance, Novartis, Merck; Medical Biosurfaces; Locus; Affinergy; Contrafect; Karius; Genentech, Regeneron, Basilea, and Janssen; received funding from Basilea, Affinergy, Janssen, Basilea, Integrated Biotherapeutics; C3J, Armata, Valanbio; Akagera, Aridis, Novartis, Novadigm, Durata, Debiopharm, Genentech, Achaogen, Affinium, Medicines Co., Cerexa, Tetraphase, Trius, MedImmune, Bayer, Theravance, Basilea, Affinergy, Janssen, xBiotech, Contrafect, Regeneron, Destiny, UpToDate; Stock options Valanbio; a patent for sepsis diagnosis (US9850539B2). Dr. McClain disclosed he has patents pending on diagnostic signatures for respiratory infections. Dr. Crisp was an employee of BioFire Diagnostics and is currently an employee of BioMérieux, Inc. Dr. Ginsburg’s institution received funding from DARPA; received support for article research from the Bill & Melinda Gates Foundation. Dr. Burke is a consultant for and holds equity in Predigen, Inc. Dr. Burke’s institution received funding from the NIH; received funding from Predigen, Inc.; disclosed he is a coinventor on patents pending on Molecular Methods to Diagnose and Treat Respiratory Infections. Dr. Hemmert disclosed the off-label product use of BioFire FilmArray System. The remaining authors have disclosed that they do not have any potential conflicts of interest., (Copyright © 2021 by the Society of Critical Care Medicine and Wolters Kluwer Health, Inc. All Rights Reserved.)

Background: The w Mel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and subsequently shown to reduce transmission of dengue and other pathogens, under both laboratory and field conditions. Here we describe the entomological outcomes of w Mel Wolbachia mosquito releases in two small communities in Nha Trang City in central Vietnam. Methods: The w Mel strain of Wolbachia was backcrossed into local Aedes aegypti genotype and mosquito releases were undertaken by community members or by staff. Field monitoring was undertaken to track Wolbachia establishment in local Ae. aegypti mosquito populations. Ecological studies were undertaken to assess relationships between environmental factors and the spatial and temporal variability in Wolbachia infection prevalence in mosquitoes. Results: Releases of w Mel Wolbachia Ae. aegypti mosquitoes in two small communities in Nha Trang City resulted in the initial establishment of Wolbachia in the local Ae. aegypti mosquito populations, followed by seasonal fluctuations in Wolbachia prevalence. There was significant small-scale spatial heterogeneity in Wolbachia infection prevalence in the Tri Nguyen Village site, resulting in the loss of w Mel Wolbachia infection in mosquitoes in north and center areas, despite Wolbachia prevalence remaining high in mosquitoes in the south area. In the second site, Vinh Luong Ward, Wolbachia has persisted at a high level in mosquitoes throughout this site despite similar seasonal fluctuations in w Mel Wolbachia prevalence. Conclusion: Seasonal variation in Wolbachia infection prevalence in mosquitoes was associated with elevated temperature conditions, and was possibly due to imperfect maternal transmission of Wolbachia . Heterogeneity in Wolbachia infection prevalence was found throughout one site, and indicates additional factors may influence Wolbachia establishment., Competing Interests: No competing interests were disclosed., (Copyright: © 2021 Hien NT et al.)

Background: Critical illness such as sepsis is a life-threatening syndrome defined as a dysregulated host response to infection and is characterized by patients exhibiting impaired immune response. In the field of diagnosis, a gap still remains in identifying the immune profile of critically ill patients in the intensive care unit (ICU)., Methods: A new multiplex immune profiling panel (IPP) prototype was assessed for its ability to semiquantify messenger RNA immune-related markers directly from blood, using the FilmArray System, in less than an hour. Samples from 30 healthy volunteers were used for the technical assessment of the IPP tool. Then the tool was clinically assessed using samples from 10 healthy volunteers and 20 septic shock patients stratified using human leukocyte antigen-DR expression on monocytes (mHLA-DR)., Results: The IPP prototype consists of 16 biomarkers that target the immune response. The majority of the assays had a linear expression with different RNA inputs and a coefficient of determination (R2) > 0.8. Results from the IPP pouch were comparable to standard quantitative polymerase chain reaction and the assays were within the limits of agreement in Bland-Altman analysis. Quantification cycle values of the target genes were normalized against reference genes and confirmed to account for the different cell count and technical variability. The clinical assessment of the IPP markers demonstrated various gene modulations that could distinctly differentiate 3 profiles: healthy volunteers, intermediate mHLA-DR septic shock patients, and low mHLA-DR septic shock patients., Conclusions: The use of IPP showed great potential for the development of a fully automated, rapid, and easy-to-use immune profiling tool. The IPP tool may be used in the future to stratify critically ill patients in the ICU according to their immune status. Such stratification will enable personalized management of patients and guide treatments to avoid secondary infections and lower mortality., (© The Author(s) 2020. Published by Oxford University Press for the Infectious Diseases Society of America.)

Vietnam is endemic for dengue. We conducted a series of retrospective and prospective studies to characterize the epidemiology of dengue and population mobility patterns in Nha Trang city, Vietnam, with a view to rational design of trials of community-level interventions. A 10-year time series of dengue case notifications showed pronounced interannual variability, as well as spatial heterogeneity in ward-level dengue incidence (median annual coefficient of variation k = 0.47). Of 451 children aged 1-10 years enrolled in a cross-sectional serosurvey, almost one-third had evidence of a past dengue virus (DENV) infection, with older children more likely to have a multitypic response indicative of past exposure to ≥ 1 serotype. All four DENV serotypes were detected in hospitalized patients during 8 months of sampling in 2015. Mobility data collected from 1,000 children and young adults via prospective travel diaries showed that, although all ages spent approximately half of their daytime hours (5:00 am-9:00 pm) at home, younger age groups (≤ 14 years) spent a significantly greater proportion of their time within 500 m of home than older respondents. Together these findings inform the rational design of future trials of dengue preventive interventions in this setting by identifying 1) children < 7 years as an optimal target group for a flavivirus-naive serological cohort, 2) children and young adults as the predominant patient population for a study with a clinical end point of symptomatic dengue, and 3) substantial spatial and temporal variations in DENV transmission, with a consequent requirement for a trial to be large enough and of long enough duration to overcome this heterogeneity.

Extension rates of a thermostable, deletion-mutant polymerase were measured from 50°C to 90°C using a fluorescence activity assay adapted for real-time PCR instruments. Substrates with a common hairpin (6-base loop and a 14-bp stem) were synthesized with different 10-base homopolymer tails. Rates for A, C, G, T, and 7-deaza-G incorporation at 75°C were 81, 150, 214, 46, and 120 seconds(-1). Rates for U were half as fast as T and did not increase with increasing concentration. Hairpin substrates with 25-base tails from 0% to 100% GC content had maximal extension rates near 60% GC and were predicted from the template sequence and mononucleotide incorporation rates to within 30% for most sequences. Addition of dimethyl sulfoxide at 7.5% increased rates to within 1% to 17% of prediction for templates with 40% to 90% GC. When secondary structure was designed into the template region, extension rates decreased. Oligonucleotide probes reduced extension rates by 65% (5'-3' exo-) and 70% (5'-3' exo+). When using a separate primer and a linear template to form a polymerase substrate, rates were dependent on both the primer melting temperature (Tm) and the annealing/extension temperature. Maximum rates were observed from Tm to Tm - 5°C with little extension by Tm + 5°C. Defining the influence of sequence and temperature on polymerase extension will enable more rapid and efficient PCR., (Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.)

Background: Radioactive DNA polymerase activity methods are cumbersome and do not provide initial extension rates. A simple extension rate assay would enable study of basic assumptions about PCR and define the limits of rapid PCR., Methods: A continuous assay that monitors DNA polymerase extension using noncovalent DNA dyes on common real-time PCR instruments was developed. Extension rates were measured in nucleotides per second per molecule of polymerase. To initiate the reaction, a nucleotide analog was heat activated at 95 °C for 5 min, the temperature decreased to 75 °C, and fluorescence monitored until substrate exhaustion in 30-90 min., Results: The assay was linear with time for over 40% of the reaction and for polymerase concentrations over a 100-fold range (1-100 pmol/L). Extension rates decreased continuously with increasing monovalent cation concentrations (lithium, sodium, potassium, cesium, and ammonium). Melting-temperature depressors had variable effects. DMSO increased rates up to 33%, whereas glycerol had little effect. Betaine, formamide, and 1,2-propanediol decreased rates with increasing concentrations. Four common noncovalent DNA dyes inhibited polymerase extension. Heat-activated nucleotide analogs were 92% activated after 5 min, and hot start DNA polymerases were 73%-90% activated after 20 min., Conclusions: Simple DNA extension rate assays can be performed on real-time PCR instruments. Activity is decreased by monovalent cations, DNA dyes, and most melting temperature depressors. Rational inclusion of PCR components on the basis of their effects on polymerase extension is likely to be useful in PCR, particularly rapid-cycle or fast PCR.

DNA polymerase activity was measured by a stopped-flow assay that monitors polymerase extension using an intercalating dye. Double-stranded DNA formation during extension of a hairpin substrate was monitored at 75°C for 2 min. Rates were determined in nucleotides per second per molecule of polymerase (nt/s) and were linear with time and polymerase concentration from 1 to 50 nM. The concentrations of 15 available polymerases were quantified and their extension rates determined in 50 mM Tris, pH 8.3, 0.5 mg/ml BSA, 2 mM MgCl₂, and 200 μM each dNTP as well as their commercially recommended buffers. Native Taq polymerases had similar extension rates of 10-45 nt/s. Three alternative polymerases showed faster speeds, including KOD (76 nt/s), Klentaq I (101 nt/s), and KAPA2G (155 nt/s). Fusion polymerases including Herculase II and Phusion were relatively slow (3-13 nt/s). The pH optimum for Klentaq extension was between 8.5 and 8.7 with no effect of Tris concentration. Activity was directly correlated to the MgCl2 concentration and inversely correlated to the KCl concentration. This continuous assay is relevant to PCR and provides accurate measurement of polymerase activity using a defined template without the need of radiolabeled substrates., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: High-resolution melting of PCR products is an efficient and analytically sensitive method to scan for sequence variation, but detected variants must still be identified. Snapback primer genotyping uses a 5' primer tail complementary to its own extension product to genotype the resulting hairpin via melting. If the 2 methods were combined to analyze the same PCR product, the residual sequencing burden could be reduced or even eliminated., Methods: The 27 exons and neighboring splice sites of the CFTR [cystic fibrosis transmembrane conductance regulator (ATP-binding cassette sub-family C, member 7)] gene were amplified by the PCR in 39 fragments. Primers included snapback tails for genotyping 7 common variants and the 23 CFTR mutations recommended for screening by the American College of Medical Genetics. After symmetric PCR, the amplicons were analyzed by high-resolution melting to scan for variants. Then, a 5-fold excess of H2O was added to each reaction to produce intramolecular hairpins for snapback genotyping by melting. Each melting step required <10 min. Of the 133 DNA samples analyzed, 51 were from CFTR patient samples or cell lines., Results: As expected, the analytical sensitivity of heterozygote detection in blinded studies was 100%. Snapback genotyping reduced the need for sequencing from 7.9% to 0.5% of PCR products; only 1 amplicon every 5 patients required sequencing to identify nonanticipated rare variants. We identified 2 previously unreported variants: c.3945A>G and c.4243-5C>T., Conclusions: CFTR analysis by sequential scanning and genotyping with snapback primers is a good match for targeted clinical genetics, for which high analytical accuracy and rapid turnaround times are important.

Evolutionary processes can interact with the mechanisms of steroid hormone action to drive interspecific variation in behavioural output, yet the exact nature of these interactions is poorly understood. To investigate this issue, we compare the endocrine machinery underlying the winner effect (an ability to increase winning behaviour in response to past victories) in two closely related species of Peromyscus mice. Typically, after winning a fight, California mice (Peromyscus californicus) experience a testosterone (T) surge that helps enhance their future winning behaviour, whereas white-footed mice (Peromyscus leucopus) experience neither a T surge nor a change in subsequent winning behaviour. However, our results indicate that when the post-victory T response of male white-footed mice is phenotypically engineered to resemble that of California mice, individuals are capable of developing a strong and lasting winner effect. Moreover, this 'induced' winner effect in white-footed mice qualitatively matches the winner effect that develops naturally in California mice. Taken together, these findings suggest that white-footed mice have the physiological machinery necessary to form a robust winner effect comparable to that formed by California mice, but are unable to endogenously activate this machinery after achieving winning experiences. We speculate that evolutionary processes, like selection, operate on the physiological substrates that govern post-victory T release to guide divergence in the winner effect between these two species.

Women who are diagnosed with breast cancer often self-administer complementary and alternative medicines to augment their conventional treatments, improve health, or prevent recurrence. Flor-Essence® herbal tonic is a complex mixture of eight herbal extracts used by cancer patients because of anecdotal evidence that it can treat or prevent disease. In this study four experimental groups of female MMTV-Neu mice were left untreated or treated with 3% Flor-Essence® in utero, from birth until 5 weeks of age, or throughout their lifetime. Palpable mammary tumor incidence and body weight was determined weekly for each group. The mice were sacrificed at 28 weeks of age and mammary tumors were enumerated to determine average tumor incidence and multiplicity for each group. Female mice exposed to Flor-Essence® herbal tonic in utero weighed significantly more than the control group (p < 0.001). The average tumor incidence and tumor multiplicity in the experimental mice treated with Flor-Essence® herbal tonic did not differ from the control animals. Flor-Essence® does not inhibit mammary tumor incidence or mammary tumor multiplicity in MMTV-Neu transgenic mice. Flor-Essence® exposure in utero causes increased body weight in experimental animals. This conclusion challenges widely available anecdotal information as well as the hopes of the consumer that this product will inhibit or suppress tumor development.

Among nucleic acid analytical methods, high-resolution melting analysis is gaining more and more attention. High-resolution melting provides simple, homogeneous solutions for variant scanning and genotyping, addressing the needs of today's overburdened laboratories with rapid turnaround times and minimal cost. The flexibility of the technique has allowed it to be adopted by a wide range of disciplines for a variety of applications. In this review we examine the broad use of high-resolution melting analysis, including gene scanning, genotyping (including small amplicon, unlabeled probe and snapback primers), sequence matching and methylation analysis. Four major application arenas are examined to demonstrate the methods and approaches commonly used in particular fields. The appropriate usage of high-resolution melting analysis is discussed in the context of known constraints, such as sample quality and quantity, with a particular focus placed on proper experimental design in order to produce successful results.

The objective was to evaluate the effects of an extended withdrawal period after feeding the beta-adrenergic agonist zilpaterol hydrochloride (ZH) for 20 d at the end of the feeding period. Three hundred eighty-four crossbred beef steers were blocked by BW and randomly allocated into 64 pens (6 steers/pen). Pens were assigned to treatments in a 2 x 4 factorial arrangement in a randomized complete block design. Main effects were the addition of 0 (control) or 8.3 mg/kg of ZH (DM basis) to the finishing diet for 20 d before estimated average slaughter date and paired withdrawal periods of 3, 10, 17, or 24 d before slaughter. Individual BW were measured initially, 1 d before ZH feeding, and 1 d before slaughter. The ZH feeding period was initiated so that control cattle in the 3-d withdrawal group would be expected to average 65% USDA Choice Quality grade and have 1.27 cm of 12th-rib fat based on visual appraisal. Carcass data were collected at slaughter. For the 3-d withdrawal steers, 2 steers from each pen were selected to determine visceral organ and total offal mass at slaughter. The ZH x withdrawal day interaction was not significant (P > 0.10) for the majority of variables. There was no difference (P > or = 0.12) due to ZH feeding for final BW, carcass-adjusted final BW, or ADG. However, DMI was decreased (P = 0.02) and G:F increased (P = 0.01) in steers fed ZH vs. control steers. As day after withdrawal of ZH increased, there was a linear increase (P < 0.001) in final BW and carcass-adjusted final BW, but a linear decrease (P < 0.001) in ADG over the finishing period and over the ZH plus withdrawal period. Overall, HCW was 380 and 369 kg (P < 0.001) for ZH and control steers, respectively. However, the difference between ZH and control was 14, 17, 5, and 6 kg with 3, 10, 17, and 24 d withdrawal, respectively (ZH x withdrawal day, P = 0.09). Feeding ZH increased dressing percentage (65.8 vs. 64.6%; P < 0.001) and LM area (94.8 vs. 89.7 cm(2); P < 0.001), and decreased calculated yield grade (2.69 vs. 2.91; P = 0.03) and percentage of cattle grading USDA Choice (31.1 vs. 42.3%; P = 0.03) compared with controls. Small intestinal mass (g/kg of empty BW) was greater (P = 0.03) for steers fed ZH compared with controls. There were no other differences (P > or = 0.11) in mass of body components, expressed in kilograms or as a fraction of empty BW. In this experiment, improvements in animal performance and HCW due to feeding ZH were generally maintained when withdrawal was extended through 10 d.

Experiments were conducted at 3 US locations (California, Idaho, and Texas) to determine the effects of dietary zilpaterol hydrochloride and duration of zilpaterol feeding on carcass composition and beef palatability. At each site, 160 steers and 160 heifers were stratified within sex by initial BW (study d -1) and assigned randomly within BW strata to 1 of 4 treatments in a randomized complete block design (4 blocks/treatment for each sex). The 4 treatments were arranged in a 2 (no zilpaterol vs. zilpaterol) x 2 (20- or 40-d duration of zilpaterol feeding) factorial. When included in the diet, zilpaterol was supplemented at 8.3 mg/kg (DM basis). Each pen consisted of 10 animals. After slaughter 2 carcasses per pen (n=64 per trial site) were selected. The entire right side of the selected carcasses was collected for dissection and chemical analysis of the soft tissue. Additionally, the left strip loin was collected for Warner-Bratzler shear force determinations and aged to 28 d postmortem. Sensory analysis was conducted on the Idaho trial site samples only. All data were pooled for analyses. Feeding zilpaterol hydrochloride increased carcass muscle deposition (P<0.01) of both steer and heifer carcasses. However, carcass percentage fat of steers and heifers was not affected (P>0.11) by the zilpaterol treatment. In heifer carcasses, carcass moisture percentage was increased (P=0.04) and bone percentage was decreased (P=0.02), whereas in steer carcasses, carcass moisture and bone percentage were not affected (P>0.10). In heifer carcasses, carcass ash percentage was not affected (P=0.61) by zilpaterol, whereas in steer carcasses, carcass ash percentage tended (P=0.07) to be increased. The protein-to-bone ratio was increased (P<0.001) by zilpaterol hydrochloride treatment in both steers and heifers, whereas the protein-to-fat ratio was not affected (P=0.10). Cooking loss of the LM was not affected (P=0.41) by zilpaterol treatment of steers or heifers. However, LM Warner-Bratzler shear force was increased (P=0.003) on average (3.3 vs. 4.0 kg) due to zilpaterol hydrochloride treatment of both steers and heifers. In both steers and heifers, LM sensory panel scores of overall juiciness (6.2 vs. 6.0), tenderness (6.2 vs. 6.0), and flavor intensity (6.2 vs. 6.0) tended (P=0.06) to be decreased in cattle supplemented with zilpaterol. Zilpaterol hydrochloride is a repartitioning agent that seems to affect carcass composition primarily through protein deposition. However, zilpaterol treatment can adversely affect tenderness and other palatability traits.

Experiments were conducted at 3 US locations (CA, ID, and TX) to determine the effects of dietary zilpaterol hydrochloride (Zilmax, Intervet Inc., Millsboro, DE) and duration of zilpaterol feeding on performance and carcass merit of finishing steers and heifers. At each site, 160 steers and 160 heifers were stratified within sex by initial BW (study d -1) and assigned randomly within BW strata to 1 of 4 treatments in a randomized complete block design (4 blocks/treatment for each sex). The 4 treatments were arranged in a 2 (no zilpaterol vs. zilpaterol) x 2 (20 or 40 d duration of zilpaterol feeding) factorial arrangement of treatments. When included in the diet, zilpaterol was supplemented at 8.3 mg/kg of DM. Each pen consisted of 10 animals. Each animal was individually weighed unshrunk on d 1, 21 or 41, and 66 of the experiment. Following d 66, cattle were slaughtered and carcass data collected. Feeding zilpaterol increased (P<0.01) final BW of steers and heifers by 11.6 and 6.7 kg, respectively. In addition, feeding zilpaterol hydrochloride increased (P or= 0.12) and KPH (P >or= 0.70) were not affected by feeding zilpaterol to steers or heifers. Feeding zilpaterol decreased (i.e., improved; P=0.02) calculated yield grade of steer and heifer carcasses. Marbling score (P=0.002) and quality grade (P=0.002) were decreased when zilpaterol hydrochloride was fed to steers, and the decrease in marbling score and quality grade tended to be greater when zilpaterol was fed for 40 compared with 20 d (zilpaterol x duration interaction, P=0.07). For heifers, marbling score tended (P=0.07) to be decreased and quality grade was decreased (P=0.05) when zilpaterol hydrochloride was fed. In general, it appears from these data that zilpaterol hydrochloride fed for 20 to 40 d at the end of the finishing period enhances growth performance and carcass muscle deposition for steers and heifers.

An experiment was conducted using 200 beef carcasses to evaluate the effects of feeding zilpaterol hydrochloride with or without monensin and tylosin on carcass cutability and meat sensory variables. The experiment was conducted using a randomized complete block design with treatments arranged as a 2 (no zilpaterol vs. zilpaterol) x 2 (monensin and tylosin withdrawn vs. monensin and tylosin fed) factorial. Cattle (n=3,757) were fed zilpaterol hydrochloride, a beta(2)-adrenergic agonist, for 30 d at the end of the finishing period and withdrawn from zilpaterol hydrochloride for the last 5 d on feed. Five carcasses (weighing between 305 and 421 kg and free of slaughter defects) were selected from each of 40 feedlot treatment pens. Strip loins from the left sides were collected for sensory analysis and Warner-Bratzler shear force (WBSF) testing, and the rib was collected for 9th, 10th, 11th-rib dissections. A subsample of 3 carcass right sides per pen was fabricated into boneless subprimals according to Institutional Meat Purchase Specifications. Carcasses from zilpaterol-fed steers had greater (P or= 0.26). For the main effect of monensin and tylosin, withdrawal of monensin and tylosin decreased (P=0.01) consumer juiciness scores, although other yield and compositional measurements were not affected (P >or= 0.07). Zilpaterol is a strong repartitioning agent that increases meat yield through increased protein and decreased fat deposition.

Forty-eight Bonsmara steers were assigned to three treatment groups and one control group consisting of 12 animals each. The control (C) received no β-agonist, while the three treatment groups received zilpaterol (6ppm) (Z), ractopamine (30ppm) (R) or clenbuterol (2ppm) (Cl) for the last thirty days on feed. Growth performance (final 30 days), USDA quality and yield grades and meat quality (shear force, chemical, histological and biochemical) were compared for the three β-agonist and control groups. Animals responded negatively to Cl treatment during initial stages of supplementation, which was evident in lower feed consumption and initial growth rates. For carcass growth and yield, Cl had greater and more efficient growth rates, higher dressed out yields (proportional), lower USDA yield grades, and reduced marbling compared with C (P<0.05). For meat quality measurements, the M. longissimus (LL) and M. semitendinosus (ST) were sampled. Cl had the greatest effect (P<0.05) on WBSF, especially on the LL, followed by Z. Variation in tenderness and ageing effects corresponded with variation in calpastatin activity and myofibrillar fragmentation between treatment groups. While zilpaterol and ractopamine are currently the only products registered for cattle in different countries, it seems that zilpaterol has an advantage in carcass growth efficiency and yield without showing any adaptation problems for animals such as experienced by the more aggressive β-agonist clenbuterol.

A feedlot experiment was conducted under commercial conditions in the Texas Panhandle using 3,757 feedlot steers (average of 94 steers/pen) to evaluate the effects of feeding zilpaterol hydrochloride with or without monensin and tylosin on feedlot performance and carcass characteristics. The experiment was conducted using a randomized complete block design. Treatments were arranged as a 2 (no zilpaterol vs. zilpaterol) x 2 (monensin and tylosin withdrawn vs. monensin and tylosin fed during the final 35 d on feed) factorial. Steers were fed for a total of 161 to 167 d, and treatments were administered during the final 35 d that cattle were on feed. When included in the diet, zilpaterol, monensin, and tylosin were supplemented at 8.3, 33.1, and 12.2 mg/kg (DM basis), respectively. Zilpaterol was included in the diet for 30 d at the end of the finishing period and withdrawn from the diet for the last 5 or 6 d cattle were on feed. Cattle were harvested and carcass data collected. There were no zilpaterol x monensin/tylosin interactions (P >or= 0.12) for ADG or G:F. Feeding zilpaterol increased ADG (P < 0.001) by 0.20 kg and G:F (P < 0.001) by 0.029 kg/kg during the last 35 d on feed. Likewise, when feedlot variables were measured throughout the entire 161- to 167-d feeding trial, ADG (3.4%; P < 0.001) and G:F (3.9%; P < 0.001) were increased. Feeding zilpaterol increased (P < 0.001) dressing percent and HCW and decreased (P < 0.001) total liver abscess rate compared with controls. In addition, zilpaterol increased (P < 0.001) LM area by an average of 8.0 cm(2). There was a zilpaterol x monensin/tylosin interaction (P = 0.03) for marbling score. Zilpaterol decreased (P < 0.001) marbling score regardless of monensin and tylosin treatment, although withdrawal of monensin and tylosin for 35 d decreased marbling to a greater extent (31 vs. 17 degrees). Zilpaterol decreased (i.e., improved; P < 0.001) calculated yield grade regardless of monensin and tylosin treatment, but feeding zilpaterol in combination with the withdrawal of monensin and tylosin for 35 d decreased calculated yield grade to a greater extent (0.49 vs. 0.29) compared with the zilpaterol, monensin, and tylosin combination treatment (zilpaterol x monensin/tylosin interaction, P = 0.03). Results suggest that monensin and tylosin can be withdrawn from the diet during the zilpaterol feeding period (final 35 d on feed) with minimal effect on animal performance, although feeding zilpaterol in combination with monensin and tylosin seemed to moderate effects on carcass quality.

Two hundred crossbred cattle (Bos indicus x Bos taurus) were supplemented with 2% Tasco (Ascophyllum nodosum) in a commercial finishing facility to evaluate marbling score, USDA quality grade, sensory traits, and retail display shelf life. Treatment animals (n = 100) received a steam-rolled corn (Zea mays)-based diet containing 2% Tasco meal (DM basis), for 14 d beginning at d 45 of the finishing period and again 14 d before slaughter. Control animals (n = 100) received a steam-rolled corn (Zea mays)-based diet without Tasco at identical feeding periods. Carcasses from Tasco-fed cattle exhibited greater marbling scores (P = 0.003) than controls. There were no treatment effects (P > 0.05) on sensory, shear, or purge attributes of striploin or inside round steaks with the exception of inside round steaks from Tasco animals having a greater initial tenderness (P = 0.03) and lower off-flavor score (P = 0.002) than control steaks. The LM samples from Tasco-fed cattle had a greater percentage of ether extractable fat (P = 0.001) and lower percentage of protein (P = 0.001) than controls. Inside round samples from treated animals exhibited a greater percentage of moisture (P = 0.03) than control steaks. Visual lean color of striploin steaks was not affected by Tasco supplementation (P = 0.26); however, steaks from Tasco-treated animals were more uniform and had less discoloration and browning than those from controls (P = 0.005, 0.04, and 0.05, respectively). Inside round visual scores and instrument values reflected similar treatment responses (P < 0.05), with a majority of the effects on muscle redness (CIE a*, hue angle) and measures of discoloration. Tasco steaks were generally more red and less discolored during extended postmortem aging and retail exposure (P < 0.05). The results from this study indicate that short-term supplementation of 2% Tasco meal in feedlot cattle increases carcass quality and prolonged retail shelf life.

Background: People diagnosed with cancer often self-administer complementary and alternative medicines (CAMs) to supplement their conventional treatments, improve health, or prevent recurrence. Flor-Essence and Essiac Herbal Tonics are commercially available complex mixtures of herbal extracts sold as dietary supplements and used by cancer patients based on anecdotal evidence that they can treat or prevent disease. In this study, we evaluated Flor-Essence and Essiac for their effects on the growth of human tumor cells in culture., Methods: The effect of Flor-Essence and Essiac((R)) herbal tonics on cell proliferation was tested in MCF-7, MDA-MB-436, MDA-MB-231, and T47D cancer cells isolated from human breast tumors. Estrogen receptor (ER) dependent activation of a luciferase reporter construct was tested in MCF-7 cells. Specific binding to the ER was tested using an ICI 182,780 competition assay., Results: Flor-Essence and Essiac herbal tonics at 1%, 2%, 4% and 8% stimulated cell proliferation relative to untreated controls in both estrogen receptor positive (MCF-7 and T47D) and estrogen receptor negative (MDA-MB-231 and MDA-MB-436) cell lines. Exposure to the tonics also produced a dose-dependent increase in ER dependent luciferase activity in MCF-7 cells. A 10(-7) M concentration of ICI 182,780 inhibited the induction of ER dependent luciferase activity by Flor-Essence and Essiac, but did not affect cell proliferation., Conclusion: Flor-Essence and Essiac Herbal Tonics can stimulate the growth of human breast cancer cells through ER mediated as well as ER independent mechanisms of action.

We use time-of-flight secondary ion mass spectrometry (TOF-SIMS) to image and classify individual cells on the basis of their characteristic mass spectra. Using statistical data reduction on the large data sets generated during TOF-SIMS analysis, similar biological materials can be differentiated on the basis of a combination of small changes in protein expression, metabolic activity and cell structure. We apply this powerful technique to image and differentiate three carcinoma-derived human breast cancer cell lines (MCF-7, T47D, and MDA-MB-231). In homogenized cells, we show the ability to differentiate the cell types as well as cellular compartments (cytosol, nuclear, and membrane). These studies illustrate the capacity of TOF-SIMS to characterize individual cells by chemical composition, which could ultimately be applied to detect and identify single aberrant cells within a normal cell population. Ultimately, we anticipate characterizing rare chemical changes that may provide clues to single cell progression within carcinogenic and metastatic pathways.

Many carcinogens have been shown to cause tissue specific tumors in animal models. The mechanism for this specificity has not been fully elucidated and is usually attributed to differences in organ metabolism. For heterocyclic amines, potent carcinogens that are formed in well-done meat, the ability to either bind to the estrogen receptor and activate or inhibit an estrogenic response will have a major impact on carcinogenicity. Here, we describe our work with the human estrogen receptor alpha (ERalpha), the mutagenic/carcinogenic heterocyclic amines PhIP, MeIQx, and IFP, and the hydroxylated metabolite of PhIP, N2-hydroxy-PhIP. We demonstrate both by computational docking and NMR analysis that PhIP binds with the ligand binding domain (LBD). This binding competes with estradiol (E2) in the native E2 binding cavity of the receptor. In vitro assays show that PhIP, in contrast to the other heterocyclic amines, increases cell proliferation in MCF-7 human breast cancer cells and activates the ERalpha receptor. We also find that other heterocyclic amines and N2-hydroxy-PhIP inhibit ERalpha activation. We propose that the mechanism for the tissue-specific carcinogenicity seen in the rat breast tumors and the presumptive human breast cancer associated with the consumption of well-done meat maybe mediated by this receptor activation.

Background: Women who are diagnosed with breast cancer often self-administer complementary and alternative medicines to augment their conventional treatments, improve health, or prevent recurrence. Flor-Essence tonic is a complex mixture of herbal extracts used by cancer patients because of anecdotal evidence that it can treat or prevent disease., Methods: Female Sprague-Dawley rats were given water or exposed to 3 or 6% Flor-Essence beginning at 1 day of age. Mammary tumors were induced with a single oral 40 mg/kg/bw dose of dimethyl-benz[a]anthracene at 50 days of age and sacrificed at 23 weeks. Rats were maintained on AIN-76A diet., Results: Control rats had palpable mammary tumor incidence of 51.0% at 19 weeks of age compared to 65.0 and 59.4% for the 3 and 6% Flor-Essence groups respectively. Overall, no significant difference in time until first palpable tumor was detected among any of the groups. At necropsy, mammary tumor incidence was 82.5% for controls compared to 90.0 and 97.3% for rats consuming 3 and 6% Flor-Essence, respectively. Mean mammary tumor multiplicity (+/-SES) for the controls was 2.8 (+/-0.5) and statistically different from the 3 or 6% Flor-Essence groups with 5.2 (+/-0.7), and 4.8 (+/-0.6), respectively (p < or = 0.01). As expected, the majority of isolated tumors were diagnosed as adenocarcinomas., Conclusions: Flor-Essence can promote mammary tumor development in the Sprague-Dawley rat model. This observation is contrary to widely available anecdotal evidence as well as the desire of the consumer that this commercially available herbal tonic will suppress and/or inhibit tumor growth.

The effect of supplementing diets with various levels of vitamin D3 to provide 0, 0.5, 1, and 5 million IU/(steer x d) for 8 d before slaughter on the mineral content and localization of Ca in LM and muscle fragments was studied during the postmortem aging process. Twelve feedlot steers of three biological types were given access to the four levels of vitamin D for 8 d before slaughter. Differential centrifugation techniques were used to determine the concentrations of minerals relative to protein in different muscle fragments on d 3 and 21 postmortem. Electron microscopy visualization of bound Ca indicated that vitamin D3 mobilized Ca from the sarcoplasmic reticulum and transverse tubule system into the myofibrils. Bound Ca was concentrated near the Z-line at the A-band/I-band juncture within the sarcomere. Supplementing steers with 1 and 5 million IU/(steer x d) of vitamin D3 increased (P < 0.05) Ca, P, and Mg concentrations per unit of protein in the cytosol. Soluble cytosolic Ca concentrations were greater (P < 0.05) on d 21 than on d 3 postmortem only when steers were supplemented with 5 million IU/d. Concentrations of Ca, P, and Mg in isolated tissues were increased (P < 0.05) in nuclei and myofibrilar proteins by supplementing steers with 1 and 5 million IU/ (steer x d) of vitamin D3. All supplemental vitamin D3 treatments also increased (P < 0.001) Mg concentrations in the cytosol, regardless of aging treatment, and increased Mg concentrations (P < 0.04) within the mitochondria at d 3 postmortem. Thus, supplementation of feedlot steers with vitamin D3 at levels of 0.5 to 5 million IU/(steer x d) increased Ca concentrations within respiring muscle, resulting in increased bound tissue Ca concentrations. When the respiring muscle was converted to meat, the increased bound tissue Ca resulting from vitamin D3 treatment released Ca concentrations into the cytosol during aging (P < 0.05). Results of this study indicate that vitamin D3 supplementation increased total cytosolic Ca, P, and Mg concentrations in meat.

Because of the Ca dependency of the calpains, oral supplementation of vitamin D3 (VITD) can increase the Ca content of muscle to activate the calpains and improve tenderness. Feedlot steers (n = 142) were arranged in a 4 x 3 factorial arrangement consisting of four levels of VITD (0, 0.5, 1, and 5 million IU/[steer x d]) for eight consecutive days antemortem using three biological types (Bos indicus, Bos taurus-Continental, and Bos taurus-English). Feedlot performance factors of ADG, DMI, and G:F were measured, and carcass quality, yield, and color data were collected. Plasma Ca and P concentrations were measured during d 4 to 6 of supplementation and at exsanguination, and carcass pH and temperature were measured in the LM at 3 and 24 h postmortem. Vitamin D3 treatment at 5 million IU/(steer x d) decreased ADG (P < 0.05) over the supplementation and feed intake for the last 2 d of feeding compared with untreated control steers. Likewise, G:F was decreased (P = 0.03) in steers supplemented with 5 million IU/d compared with controls. Overall, there was a linear decrease (P < 0.01) in ADG and G:F as a result of VITD supplementation. Plasma concentrations of Ca and P were increased (P < 0.05) by VITD concentrations of 1 and 5 million IU/(steer x d). All VITD treatments increased (P < 0.05) LM temperature at 3 h postmortem and pH at 24 h postmortem. Vitamin D3 treatments did not affect (P = 0.07) any other carcass measurements, including USDA yield and quality grade; thus, any improvements in meat tenderness as a result of VITD supplementation can be made without adversely affecting economically important carcass factors. Biological type of cattle did not interact with VITD treatment for any carcass or feedlot performance trait. Although feeding 5 million IU/(steer x d) of VITD for eight consecutive days had negative effects on performance, supplementing VITD at 0.5 million IU/ (steer x d) did not significantly alter feedlot performance.

Vitamin D3 was orally supplemented to determine the supplemental dose that improved beef tenderness in different cattle breed types. Feedlot steers (n = 142) were arranged in a 4 x 3 factorial arrangement consisting of four levels of supplemental vitamin D3 (0, 0.5, 1, and 5 million IU/steer daily) administered for eight consecutive days antemortem using three biological types (Bos indicus, Bos Taurus-Continental, and Bos Taurus-English). Warner-Bratzler shear force (WBSF) was measured at 3, 7, 10, 14, and 21 d postmortem, and trained sensory analysis was conducted at 7 d postmortem on LM, semimembranosus, gluteus medius, and supraspinatus steaks. Concentrations of vitamin D3 and the metabolites 25-hydroxyvitamin D3, and 1,25-dihydroxyvitamin D3 were determined in the LM, liver, kidney, and plasma. Biological type of cattle did not interact (P > 0.10) with vitamin D3 supplementation for sensory or tenderness traits, suggesting that feeding vitamin D3 for 8 d before slaughter affected the different biological types of cattle similarly. Supplementing steers with 0.5, 1, or 5 million IU/(steer(d) decreased (P < 0.05) LM WBSF at 7, 10, 14, and 21 d postmortem compared with controls, and vitamin D3 treatments of 0.5, 1, and 5 million IU decreased (P < 0.05) semimembranosus WBSF at 3, 7, and 14 d postmortem. In general, vitamin D3-induced improvements in WBSF were most consistent and intense in LM steaks. Sensory panel tenderness was improved (P < 0.05) by all vitamin D3 treatments in LM steaks. Sensory traits ofjuiciness, flavor, connective tissue, and off-flavor were not (P > 0.05) affected by vitamin D3 treatments. All vitamin D3 treatments decreased micro-calpain activity and increased muscle Ca concentrations (P < 0.05). Vitamin D3 concentrations were increased (P < 0.05) by supplementation in all tissues tested (liver, kidney, LM, and plasma); however, cooking steaks to 71 degrees C decreased (P < 0.05) treatment residue effects. The vitamin D metabolite 1,25-dihydroxyvitamin D3 was increased (P < 0.05) only in plasma samples as a result of the vitamin D3 treatments. These results indicate that supplementation with vitamin D3 at 0.5 million IU/steer daily for eight consecutive days before slaughter improved tenderness in steaks from different subprimal cuts by affecting muscle Ca concentrations, micro-calpain activities, and muscle proteolysis, with only a small effect on tissue residues of vitamin D3.

Feedlot steers (n = 36) from three biological types (Bos indicus, Bos taurus-Continental, and Bos taurus-English) were used to determine the Ca, P, and vitamin D3 status of feedlot cattle. The USDA yield and quality grade traits were measured at slaughter, and the concentrations of vitamin D3 (VITD) and the metabolites 25-hydroxyvitamin D3 (25-OH D) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D) were determined in LM, liver, kidney, and plasma. Plasma and muscle Ca and P concentrations also were determined. Biological type of cattle affected a number of carcass traits. Carcasses from Bos taurus-English cattle had more marbling, resulting in higher quality grades (P < 0.05). Carcasses from Bos taurus-Continental cattle had lower calculated yield grades (P < 0.05) than did carcasses from cattle in the other biological types. In general, differences in carcass traits resulting from biological type were consistent with other reports. Plasma and LM Ca and P concentrations were not affected (P = 0.06) by biological type of cattle, indicating that Ca and P homeostasis is a conserved trait across the different types of cattle. Plasma VITD and 25-OH D concentrations were not affected (P = 0.41) by biological type, whereas plasma 1,25-(OH)2 D concentration was lower (P < 0.05) in Bos taurus-English cattle than in Bos taurus-Continental and Bos indicus cattle. Liver VITD and 25-OH D were not affected by biological type (P = 0.76), but liver 1,25-(OH)2 D concentration was greater (P < 0.05) in Bos indicus cattle than in Bos taurus-Continental cattle. Kidney vitamin D metabolite concentrations were not affected by biological type of cattle (P = 0.21). Muscle VITD concentration was greater (P < 0.05) in Bos taurus-English cattle than in the other two biological types, and muscle 25-OH D concentrations were greater (P < 0.05) in Bos taurus-English cattle than in Bos indicus cattle. Muscle 1,25-(OH)2 D concentration was less (P < 0.05) in the Bos taurus-Continental cattle than in the other two biological types. Cooking eliminated vitamin D metabolite differences among the biological types. Our results suggest that Bos indicus cattle had greater 1,25-(OH)2 D (the biologically active form) in tissues, and greater 1,25-(OH)2 D plasma concentrations than Bos taurus cattle. Thus, the need for VITD supplementation and optimal levels of Ca and P in feedlot diets might differ between Bos indicus and Bos taurus cattle.

Effects of irradiation (2 kGy) of ground beef patties from trimmings stored aerobically for 0 or 6 days on lean color, odor, and sensory attributes were investigated. Beef trimings were coarse ground and split into 2 groups. Group one was fine ground, pattied, and packaged immediately; group-two was stored 6 days then fine ground, pattied, and packaged. Irradiated beef patties had greater (P<0.05) off-odors, and off-flavors, lower (P<0.05) CIE L(∗), a(∗) and b(∗) and saturation indexes values (P<0.05) after four days of storage at 0±1 °C. Irradiation of patties produced from trimmings aged an extra 6 days resulted in increased (P<0.05) saturation indexes and b(∗) values, but not off-odors when compared to non-aged and irradiated patties. Thus, the production of irradiated beef patties should utilize beef trimmings with the shortest postmortem aging time and a dose of less than 2 kGy to minimize discoloration and off-odors.

Primary bovine muscle cell culture studies were conducted to determine whether implanting heifers had a direct effect on in vitro protein synthesis and degradation and to determine the effect of implanting heifers on longissimus muscle palatability. Feedlot heifers (n = 96) were administered one of six implant regimens to characterize their effect on in vitro amino acid uptake and protein degradation. Treatments consisted of: 1) a nonimplanted control (NI/NI); 2) no implant on d 1 and Revalor-H administered on d 84 of the experiment (NI/Rev); 3) Revalor-H on d 1, but no implant given at d 84 (Rev/NI); 4) Revalor-H administered on d 1 and d 84 (Rev/Rev); 5) Revalor-IH administered on d 1 and Revalor-H at d 84 (RIH/Rev); and 6) Synovex-H given at d 1 and Revalor-H administered at d 84 (Syn/Rev). Blood and longissimus lumborum muscle were collected 20 min postmortem, and serum and muscle extracts were incubated with primary bovine muscle cells. Implant treatments had minimal effects on shear force and sensory traits; however, steaks from Rev/Rev heifers were 0.31 kg more tender (P < 0.05) than steaks from NI/NI heifers. Serum protein synthesis and degradation were not affected (P > 0.10) by any implant treatment. When primary bovine muscle cells were treated with muscle extract, amino acid uptake was greater for heifers implanted with Rev/ Rev than for the average of all other treatments (P < 0.01). The Rev/Rev implant regimen also increased (P < 0.05) amino acid uptake compared with heifers treated with RIH/Rev, Syn/Rev, NI/NI, NI/Rev, or Rev/NI. Cellular protein degradation of the muscle cell culture treated with muscle extract tended (P < 0.10) to be higher in NI/NI-treated cells compared with the average of all implant treatments. In addition, cells treated with muscle extract from heifers implanted with Rev/Rev had lower (P < 0.05) protein degradation than the NI/NI control heifers. These results indicate that anabolic implant strategies can directly affect both muscle protein synthesis and degradation via effects that seem to be more autocrine than paracrine in nature.

Boneless top loin subprimals (n = 320) from Slight and Small marbled carcasses were fabricated into 2.54-cm thick steaks to determine core location effects on tenderness. In Exp. 1, top loins were aged to 7 d before steaks were cut and cooked to an internal temperature of 71 degrees C. After cooking, a maximum of 15 1.27-cm diameter cores were removed and sheared with a Warner-Bratzler shear force (WBSF) device. There was not a marbling score x core location interaction (P = 0.36). However, there was a main effect of core location (P < 0.01). Cores from the medial, middle, and lateral portion of the longissimus muscle (LM) aged for 7 d differed, with less resistance (P < 0.05) in the medial than the lateral end. Also, there was an effect of marbling score on WBSF, with Small-marbled steaks having lower (P < 0.02) WBSF values than Slight-marbled steaks. In a second experiment, steaks were removed from the middle of the top loin subprimals and aged an additional 7 d to produce 14-d aged steaks. Shear values decreased (P < 0.05) from Exp. 1 to 2 for all core locations. Neither the main effect of marbling score nor the core location x marbling score interaction was significant (P > 0.40); however, the same lateral to medial gradient in WBSF values was discovered again in Exp. 2. Both experiments indicated there were regions of WBSF values that differed (P < 0.05) across the cross section of the LM producing a shear-force/tenderness gradient, with the most medial cores having the lowest WBSF values in both experiments independent of marbling score. Regression analyses indicated the middle and center portions of LM steaks tended to have the most predictive capacity of average WBSF. Because of the variability in tenderness caused by location within the LM, care should be exercised when selecting sampling areas for the measurements of tenderness using the WBSF measure.

The objective of this experiment was to determine the effect of different doses of vitamin D3 (VITD) on beef feedlot performance, plasma and muscle Ca2+, tissue residues, and improvement of Warner-Bratzler shear force (WBS) and panel tenderness. A total of 167 steers were fed one of six levels of VITD. The VITD treatments (28 steers/treatment) were 0, 0.5 x 10(6), 1 x 10(6), 2.5 x 10(6), 5 x 10(6), and 7.5 x 10(6) IU/steer daily of VITD fed nine consecutive days before slaughter. Feedlot performance and plasma Ca2+ were measured during the last 21 days on feed. Warner-Bratzler shear force was measured on strip loin and top round steaks at 7, 10, 14, and 21 d postmortem. The VITD treatments of 5 and 7.5 x 10(6) IU/steer daily decreased (P < 0.05) ADG, and VITD supplementation of 2.5, 5, and 7.5 x 10(6) IU/steer daily decreased average dry matter feed intake (P < 0.05) at the end of the feeding trial. Plasma Ca2+ increased linearly with VITD treatment (P < 0.01). Calpastatin and calpain activity were not influenced by treatment (P > 0.05), but muscle Ca2+ was increased (P < 0.05) by VITD treatments of 1, 2.5, 5, and 7.5 10(6) IU/steer daily. Feeding VITD did not influence (P > 0.05) carcass quality or yield traits. Supplementing VITD at levels of 1, 2.5, 5, and 7.5 10(6) IU/steer daily increased (P < 0.05) VITD concentrations in strip loin and liver samples. Cooking liver decreased VITD concentrations 10 to 28%. Vitamin D3 treatments of 0.5 and 7.5 x 10(6) IU/d reduced strip loin steak WBS at d 7 (P < 0.05), but VITD treatments did not decrease strip loin steak WBS at any other time postmortem. The VITD treatments of 0.5, 1, and 5 x 10(6) IU/steer daily decreased top round steak WBS at 7 d, and all VITD treatments decreased 10-d top round steak WBS (P < 0.05). Supplementing steers with 0.5 x 10(6) IU/steer daily of VITD also decreased (P < 0.05) top round steak WBS at 21 d postmortem compared with controls. Sensory tenderness at 7 d postmortem was increased (P < 0.05) by all VITD treatments in top round steaks, yet strip loin tenderness scores were not affected (P > 0.05) by VITD treatment. Treatment with VITD quadratically decreased (P < 0.05) round WBS. Thus, VITD treatment will effectively improve tenderness when cattle tend to be tough and have no impact on cattle that produce tender beef. Feeding steers 0.5 x 10(6) IU of VITD daily for 9 d improved tenderness in two muscles without negatively affecting feedlot performance or tissue residues.

A detailed methodology is described for determination of treatment effects on muscle cell protein synthesis and muscle cell protein degradation in a cell culture system. C(2)C(22) mouse myoblasts were treated with growth media containing muscle extracts from bovine treated with different pharmaceutical agents. Radiolabeled amino acids were added to the growth media to determine treatment effects on protein synthesis and protein degradation. Percent protein synthesis was calculated by measuring amino acid uptake as a percentage over internal control. Percent protein degradation was measured using a pulse chase technique. These procedures will allow researchers to determine treatment effects on overall protein synthesis and degradation in vitro in a relatively short amount of time without excessive costs. A second benefit is that animals do not have to be fed radiolabeled feedstuffs. These procedures are not intended to elucidate the mechanisms behind pharmaceutical enhancement of muscle cell protein synthesis or protein degradation.

Tall fescue (Festuca arundinacea Schreb.) infected with the endophyte Neotyphodium coenophialum ([Morgan-Jones and Gams] Glenn, Bacon, and Hanlin) causes fescue toxicosis in cattle grazing the forage, but effects of the endophyte were considered to be abated soon after removal of the animals from pastures. Tasco-Forage, a proprietary extract from the brown seaweed Ascophyllum nodosum, is a known source of cytokinins and has increased antioxidant activity in both plants and the animals that graze the forage. Tasco was applied at 0 and 3.4 kg/ha to infected and uninfected tall fescue pastures in Virginia and Mississippi. Forty-eight steers grazed the pastures at each location during each of 2 yr (n = 192) before being transported to Texas for feedlot finishing. On arrival at the feedlot, steers from Tasco-treated pastures had higher (P < 0.01) monocyte phagocytic activity and tended (P < 0.07) to have higher major histocompatibility complex class II expression than steers that grazed the untreated pastures. A depression (P < 0.05) in monocyte immune cell function due to grazing infected fescue was detected throughout the feedlot finishing period but was reversed by Tasco. Rectal temperatures were elevated (P < 0.07) in steers that had grazed the infected tall fescue when they arrived in Texas, but by d 14 no difference was detected. However, by d 28 the temperature effects of infected tall fescue were reversed. Steers that had grazed infected fescue had lower (P < 0.01) rectal temperatures on d 112 of the feedlot period, demonstrating a much longer-lasting effect of the endophyte on thermoregulatory mechanisms than previously thought. Steers that had grazed Tasco-treated pastures had higher (P < 0.01) rectal temperatures on d 56 than steers that had grazed untreated fescue. Steers that had grazed the Tasco-treated pastures had higher marbling scores (P < 0.05) regardless of the endophyte, but no effect of Tasco or endophyte on gain was measured. Our data suggest that Tasco application to tall fescue pastures alleviated some of the negative effects of tall fescue toxicity.

Tasco-Forage is an Ascophyllum nodosum seaweed-based product that has increased antioxidant activity in both plants and animals. Endophyte (Neotyphodium coenophialum ([Morgan-Jones and Gams] Glenn, Bacon, and Hanlin)-infected and uninfected tall fescue (Festuca arundinacea Schreb.) pastures in Virginia and Mississippi during 1997 were treated or not with 3.4 kg Tasco/ha in April and July. There were two replications of each treatment at each location. Forty-eight steers (6/replication) grazed pastures at each location (n = 96) from April to October prior to transportation to Texas Tech, Lubbock, for finishing during a 160-d period in the feedlot. Blood (antemortem) and liver (postmortem) samples were collected. After slaughter and chilling, the left strip loins (IMPS #180) were collected from three randomly selected steers from within each pasture replication (n = 48). Strip loins were vacuum-packaged and stored at 2 degrees C. At postmortem d 7, 14, 21, and 28, strip loins were removed from packaging and fabricated into 2.54-cm steaks. Following each fabrication day postmortem, the strip loins were repackaged and stored at 2 degrees C until the following postmortem time. After the prescribed fabrication, steaks were overwrapped with polyvinyl chloride film, subjected to simulated retail display at 2 degrees C for up to 3 d, and subjective and objective color were evaluated daily by a trained panel. Steaks from Mississippi steers that had grazed Tasco-treated fescue retained higher (P < 0.05) CIE a* color scores throughout retail display. Steaks were more uniform and had less discoloration and less browning (P < 0.05) if they were from steers that had grazed Tasco-treated fescue, and the effect was greatest for steers from Mississippi (location x Tasco interaction; P < 0.05). The endophyte in tall fescue may decrease uniformity and increase lean discoloration and two-toning of beef steaks when removed from vacuum packaging on or beyond d 21 postmortem (endophyte x Tasco x postmortem day interaction: P < 0.05). Vitamin E in liver was increased (P < 0.06) and serum vitamin E was decreased (P < 0.09) in steers that had grazed the treated pastures. These experiments indicated that Tasco applied to tall fescue during the grazing season can improve color stability and extend beef shelf-life, particularly in cattle grazing infected tall fescue. The mode of action of Tasco is not clear, but antioxidants and specific vitamins may be involved.

An experiment was designed to test the hypothesis that short-term oral administration of dietary vitamin D3 to beef cattle before slaughter would increase beef tenderness through greater calcium-activated calpain activity in postmortem aged skeletal muscle. Thirty continental crossbred steers were allotted randomly to three treatment groups housed in one pen. One group served as a control; two other groups were administered boluses with either 5 x 10(6) or 7.5 x 10(6) IU of vitamin D3 daily for 9 d. Cattle were slaughtered 1 d later. The longissimus lumborum was excised from each carcass 72 h postmortem and steaks removed at 3, 7, 14, and 21 d postmortem. The semimembranosus muscle (top round) was excised from each carcass 72 h postmortem and steaks removed at 7, 14, and 21 d postmortem. Blood plasma calcium concentration of cattle treated with 5 or 7.5 x 10(6) IU of vitamin D3 was higher (P < .05) than that of controls. Strip loin and top loin steaks from cattle fed supplemental doses of vitamin D3 had lower (P < .05) Warner-Bratzler (W-B) shear values at 14 d postmortem but were not significantly different from controls at 3, 7, or 21 d (strip loins) or 7 or 21 d (top rounds). No significant difference in strip loin steak tenderness was observed by sensory panel at 14 d postmortem (P < .17) between steaks from control and vitamin D3-treated steers. At 14 d postmortem, strip loin and top round steaks from cattle fed 5 x 10(6) IU of vitamin D3, but not from those given 7.5 x 10(6) IU, showed more proteolysis (P < .05) than did steaks from control cattle, based on Western blotting analysis. Therefore, the use of supplemental dietary vitamin D3 given daily for 9 d before slaughter did improve tenderness (lower W-B shear values) of 14-d postmortem aged beef. Increased proteolysis seems to be the mechanism of tenderization.

Effects of preslaughter rest (0, 1, 2 or 3 h) and hot-fat trimming (HFT, one side) on muscle quality characteristics of hams and loins were tested with 120 pigs. Center-cut loins from each carcass side were vacuum packaged and stored for 7 or 14 days at 2°C. Pigs not rested before slaughter had a skin temperature 3°C higher and ham muscle temperatures 1°C higher than pigs rested 1-3 h (P<0.05). Three hour muscle pH was highest if pigs were rested for 1 h. HFT significantly accelerated carcass chilling rate, but did not affect rate of muscle pH decline. After 14 days of aging, pigs not rested had lower sensory panel ratings for juiciness, tenderness, flavor and overall mouthfeel of loin chops than pigs rested 1 or 2 h. Percentage bound water of hams was highest (P<0.05) if pigs rested 1 or 2 h rather than a shorter or longer times. Pigs should be rested 1 h before slaughter and HFT carried out to allow more rapid carcass chilling and improve muscle quality.

It has been known for almost a decade and a half that in trypanosomes all mRNAs are trans-spliced by addition to the 5' end of the spliced leader (SL) sequence. During the same time period the conviction developed that classical cis-splicing introns are not present in the trypanosome genome and that the trypanosome gene arrangement is highly compact with small intergenic regions separating one gene from the next. We have now discovered that these tenets are no longer true. Poly(A) polymerase (PAP) genes in Trypanosoma brucei and Trypanosoma cruzi are split by intervening sequences of 653 and 302 nt, respectively. The intervening sequences occur at identical positions in both organisms and obey the GT/AG rule of cis-splicing introns. PAP mRNAs are trans-spliced at the very 5' end as well as internally at the 3' splice site of the intervening sequence. Interestingly, 11 nucleotide positions past the actual 5' splice site are conserved between the T. bruceiand T. cruzi introns. Point mutations in these conserved positions, as well as in the AG dinucleotide of the 3' splice site, abolish intron removal in vivo. Our results, together with the recent discovery of cis-splicing introns in Euglena gracilis, suggest that both trans- and cis-splicing are ancient acquisitions of the eukaryotic cell.

Preview A patient with a suspected cervical spine injury must be handled with the utmost care until fractures and other causes of instability have been excluded. In this article, the authors describe essential imaging techniques and discuss the evaluation of common- and some uncommon- injuries.

The drug genistein, a tyrosine (Tyr) kinase inhibitor, was used to define a role for Tyr phosphorylation in regulation of basal and stimulated neutral NaCl absorption in rabbit ileum. Brush-border vesicles contain Tyr-phosphorylated peptides. Genistein freeze-thawed into the vesicles caused a concentration-dependent inhibition of at least three peptides with M(r) 111,000, 83,000, and 80,000. Studied with the Ussing chamber-voltage clamp technique, genistein added to the ileal mucosal surface inhibited neutral NaCl absorption. Direct addition of genistein to brush-border vesicles made from ileal villus cells inhibited brush-border Na(+)-H+ exchange but not D-glucose-stimulated Na+ uptake. These effects were not duplicated by genistin, a drug with similar structure to genistein but lacking Tyr kinase inhibiting properties. Serosal but not mucosal epidermal growth factor (EGF) stimulated NaCl absorption. Mucosal genistein but not genistin also altered second-messenger regulation of neutral NaCl absorption, inhibiting the effect of Ca2+ ionophore A-23187 and of serosal EGF but not affecting the transport changes caused by 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). In contrast, the Cl secretory effects indicated by the increase in short-circuit current for all three agents, A-23187, EGF, and 8-BrcAMP, were inhibited by mucosal genistein. These results suggest that 1) a Tyr kinase is involved in basally stimulating ileal neutral NaCl absorption and brush-border Na(+)-H+ exchange; 2) EGF stimulates NaCl absorption by an effect exerted from the serosal surface, but the effect also involves a brush-border Tyr kinase; 3) brush-border Tyr kinase is involved in the ability of Ca2+ ionophore A-23187 to inhibit neutral NaCl absorption but is not involved in the transport effects of cAMP. This study suggests that Tyr kinase(s) acting over short time periods is involved in stimulation of neutral NaCl absorption and brush-border Na(+)-H+ exchange and also in Ca(2+)-induced inhibition of NaCl absorption. These studies represent the first example of a brush-border Tyr kinase being involved in short-term signal transduction in intestinal epithelial cells.