The colour and colour stability of beef Longissimus dorsi and Semimembranosus muscles after effective electrical stimulation

The influence of effective low voltage electrical stimulation (ES) of beef on the colour and colour stability of longissimus dorsi (LD) and semimembranosus (SM) muscles, during storage and retail display was studied by tristimulus colorimetry and reflectance spectrophotometry. ES had no significant effect on the colour of the LD muscles, but some significant effects on SM muscles of ultimate pH 5·5-5·7. Three hours after slicing into steaks at 5 days post mortem, stimulated SM muscles had a paler/lighter colour than non-stimulated controls. During a retail display of 3 days, all steaks exhibited a loss of colour quality manifested in loss of redness and decreases in both hue and chroma (saturation). These changes were most marked in the stimulated SM muscles, and analysis indicated that they were due almost exclusively to the formation of metmyoglobin (metMb). Ageing the meat, as primals cuts, for 33 days at 0°C led to no significant differences in the perceived colour three hours after slicing. The colour changes observed during the 3-day retail display of steaks occurred more rapidly in both (ES and non-ES) 33 day-aged samples than in the 5 day-aged ones. The result of this was that the colour stability of non-stimulated steaks prepared at 33 days was similar to that of ES steaks prepared fresh (5 day post mortem). In SM muscles of pH 5·8-6·0 the apparent differences in colour of the ES and non-ES samples were not significant. However, meat of pH > 5·8, although darker than meat of lesser pH, had less tendency to form metmyoglobin during retail display.

The present work also confirmed that seemingly small differences in display conditions, especially temperature, have a marked effect on metmyoglobin formation.     

Fecal shedding of Salmonella spp. by dairy cows on farm and at cull cow markets

As part of a national study of the U.S. dairy cow population, fecal samples were collected from representative cows on 91 dairies and 97 cull dairy cow markets in 19 states. Salmonella spp. were recovered from 5.4% of milk cows, 18.1% of milk cows expected to be culled within 7 days, and 14.9% of culled dairy cows at markets. On a premise basis, Salmonella shedding in milk cows was detected on 21.1% of dairies and 66% of cull dairy cow markets. The percentage of herds with at least one cow with detectable Salmonella fecal shedding was higher during the sampling period from May through July, in herds with at least 100 milk cows, and in herds in the South region. The most common Salmonella serogroups isolated were E (30.8% of isolates) and C1 (28.6%); the most common serotypes isolated were Salmonella Montevideo (21.5% of isolates), Salmonella Cerro (13.3%), and Salmonella Kentucky (8.5%). Fecal shedding of Salmonella Typhimurium or Salmonella Typhimurium var. copenhagen was infrequent (2.8% of isolates). Most isolates (88.9%) were susceptible to all 17 antimicrobials evaluated; multiple resistance was an infrequent occurrence. This study provides information describing the distribution of Salmonella fecal shedding from dairy cows on farm and at markets and will serve as a baseline for future studies.     

Electrostatics at the nanoscale

Electrostatic forces are amongst the most versatile interactions to mediate the assembly of nanostructured materials. Depending on experimental conditions, these forces can be long- or short-ranged, can be either attractive or repulsive, and their directionality can be controlled by the shapes of the charged nano-objects. This Review is intended to serve as a primer for experimentalists curious about the fundamentals of nanoscale electrostatics and for theorists wishing to learn about recent experimental advances in the field. Accordingly, the first portion introduces the theoretical models of electrostatic double layers and derives electrostatic interaction potentials applicable to particles of different sizes and/or shapes and under different experimental conditions. This discussion is followed by the review of the key experimental systems in which electrostatic interactions are operative. Examples include electroactive and "switchable" nanoparticles, mixtures of charged nanoparticles, nanoparticle chains, sheets, coatings, crystals, and crystals-within-crystals. Applications of these and other structures in chemical sensing and amplification are also illustrated.     

Distinguishing Defensive Characteristics in the Phloem of Ash Species Resistant and Susceptible to Emerald Ash Borer

We examined the extent to which three Fraxinus cultivars and a wild population that vary in their resistance to Emerald Ash Borer (EAB) could be differentiated on the basis of a suite of constitutive chemical defense traits in phloem extracts. The EAB-resistant Manchurian ash (F. mandshurica, cv. Mancana) was characterized by having a rapid rate of wound browning, a high soluble protein concentration, low trypsin inhibitor activities, and intermediate levels of peroxidase activity and total soluble phenolic concentration. The EAB-susceptible white ash (F. americana, cv. Autumn Purple) was characterized by a slow wound browning rate and low levels of peroxidase activity and total soluble phenolic concentrations. An EAB-susceptible green ash cultivar (F. pennsylvanica, cv. Patmore) and a wild accession were similar to each other on the basis of several chemical defense traits, and were characterized by high activities of peroxidase and trypsin inhibitor, a high total soluble phenolic concentration, and an intermediate rate of wound browning. Lignin concentration and polyphenol oxidase activities did not differentiate resistant and susceptible species. Of 33 phenolic compounds separated by HPLC and meeting a minimum criterion for analysis, nine were unique to Manchurian ash, five were shared among all species, and four were found in North American ashes and not in the Manchurian ash. Principal components analysis revealed clear separations between Manchurian, white, and green ashes on the basis of all phenolics, as well as clear separations on the basis of quantities of phenolics that all species shared. Variation in some of these constitutive chemical defense traits may contribute to variation in resistance to EAB in these species.     

Noncanonical amino acids in the interrogation of cellular protein synthesis

Proteins in living cells can be made receptive to bioorthogonal chemistries through metabolic labeling with appropriately designed noncanonical amino acids (ncAAs). In the simplest approach to metabolic labeling, an amino acid analog replaces one of the natural amino acids specified by the protein's gene (or genes) of interest. Through manipulation of experimental conditions, the extent of the replacement can be adjusted. This approach, often termed residue-specific incorporation, allows the ncAA to be incorporated in controlled proportions into positions normally occupied by the natural amino acid residue. For a protein to be labeled in this way with an ncAA, it must fulfill just two requirements: (i) the corresponding natural amino acid must be encoded within the sequence of the protein at the genetic level, and (ii) the protein must be expressed while the ncAA is in the cell. Because this approach permits labeling of proteins throughout the cell, it has enabled us to develop strategies to track cellular protein synthesis by tagging proteins with reactive ncAAs. In procedures similar to isotopic labeling, translationally active ncAAs are incorporated into proteins during a "pulse" in which newly synthesized proteins are tagged. The set of tagged proteins can be distinguished from those made before the pulse by bioorthogonally ligating the ncAA side chain to probes that permit detection, isolation, and visualization of the labeled proteins. Noncanonical amino acids with side chains containing azide, alkyne, or alkene groups have been especially useful in experiments of this kind. They have been incorporated into proteins in the form of methionine analogs that are substrates for the natural translational machinery. The selectivity of the method can be enhanced through the use of mutant aminoacyl tRNA synthetases (aaRSs) that permit incorporation of ncAAs not used by the endogenous biomachinery. Through expression of mutant aaRSs, proteins can be tagged with other useful ncAAs, including analogs that contain ketones or aryl halides. High-throughput screening strategies can identify aaRS variants that activate a wide range of ncAAs. Controlled expression of mutant synthetases has been combined with ncAA tagging to permit cell-selective metabolic labeling of proteins. Expression of a mutant synthetase in a portion of cells within a complex cellular mixture restricts labeling to that subset of cells. Proteins synthesized in cells not expressing the synthetase are neither labeled nor detected. In multicellular environments, this approach permits the identification of the cellular origins of labeled proteins. In this Account, we summarize the tools and strategies that have been developed for interrogating cellular protein synthesis through residue-specific tagging with ncAAs. We describe the chemical and genetic components of ncAA-tagging strategies and discuss how these methods are being used in chemical biology.     

A hierarchical zero-inflated log-normal model for skewed responses

Although considerable attention has been given to zero-inflated count data, research on zero-inflated lognormal data is limited. In this article, we consider a study to examine human sperm cell DNA damage obtained from single-cell electrophoresis (COMET assay) experiment in which the outcome measures present a typical example of log-normal data with excess zeros. The problem is further complicated by the fact that each study subject has multiple outcomes at each of up to three visits separated by six-week intervals. Previous methods for zero-inflated log-normal data are based on either simple experimental designs, where comparison of means of zero-inflated log-normal data across different experiment groups is of primary interest, or longitudinal measurements, where only one observation is available for each subject at each visit. Their methods cannot be applied when multiple observations per visit are possible and both inter- and intra-subject variations are present. Our zero-inflated model extends the previous methods by incorporating a hierarchical structure using latent random variables to take into account both inter- and intra-subject variations in zero-inflated log-normal data. An EM algorithm has been developed to obtain the Maximum likelihood estimates of the parameters and their standard errors can be estimated by parametric bootstrap. The model is illustrated using the COMET assay data.     

Parallel Rosenbrock methods for chemical systems

Parallel Rosenbrock methods are developed for systems with stiff chemical reactions. Unlike classical Runge-Kutta methods, these linearly implicit schemes avoid the necessity to iterate at each time step. Parallelism across the method allows for the solution of the linear algebraic systems in essentially Backward Euler-like solves on concurrent processors. In addition to possessing excellent stability properties, these methods are computationally efficient while preserving positivity of the solutions. Numerical results confirm these characteristics when applied to problems involving stiff chemistry, and enzyme kinetics.     

trans-10, cis-12 conjugated linoleic acid decreases lipogenic rates and expression of genes involved in milk lipid synthesis in dairy cows

Feeding conjugated linoleic acid (CLA) reduces milk fat synthesis in lactating dairy cows, and the effect has been shown to be specific for the trans-10, cis-12 CLA isomer. Our objectives were to examine potential mechanisms by which trans-10, cis-12 CLA inhibits milk fat synthesis. Multiparous Holstein cows (n = 4) in late lactation were used in a balanced 2 x 2 crossover design. Treatments consisted of a 5 d abomasal infusion of either skim milk (control) or purified trans-10, cis-12 CLA (13.6 g/d) emulsified in skim milk. On d 5 of infusion, mammary gland biopsies were performed and a portion of the tissue analyzed for mRNA expression of acetyl CoA carboxylase, fatty acid synthetase, delta 9-desaturase, lipoprotein lipase, fatty acid binding protein, glycerol phosphate acyltransferase and acylglycerol phosphate acyltransferase. Lipogenic capacity was evaluated with another portion of the tissue. Infusion of trans-10, cis-12 CLA decreased milk fat content and yield 42 and 48%, respectively and increased the trans-10, cis-12 CLA content in milk fat from < 0.1 to 4.9 mg/g. Reductions in milk fat content of C4 to C16 fatty acids contributed 63% to the total decrease in milk fat yield (molar basis). Analysis of the ratios of specific fatty acid pairs indicated trans-10, cis-12 CLA also shifted fatty acid composition in a manner consistent with a reduction in delta 9-desaturase. Mammary explant incubations with radiolabeled acetate established that lipogenic capacity was decreased 82% and acetate oxidation to CO2 was reduced 61% when cows received trans-10, cis-12 CLA. Infusing trans-10, cis-12 CLA also decreased the mRNA expression of all measured enzymes by 39 to 54%. Overall, data demonstrated the mechanism by which trans-10, cis-12 CLA inhibits milk fat synthesis includes decreasing expression of genes that encode for enzyme involved in circulating fatty acid uptake and transport, de novo fatty acid synthesis, desaturation of fatty acids and triglyceride synthesis.     

Nitrogen isotopes as indicators of NO(x) source contributions to atmospheric nitrate deposition across the midwestern and northeastern United States

Global inputs of NO(x) are dominated by fossil fuel combustion from both stationary and vehicular sources and far exceed natural NO(x) sources. However, elucidating NO(x) sources to any given location remains a difficult challenge, despite the need for this information to develop sound regulatory and mitigation strategies. We present results from a regional-scale study of nitrogen isotopes (delta15N) in wet nitrate deposition across 33 sites in the midwestern and northeastern U.S. We demonstrate that spatial variations in delta15N are strongly correlated with NO(x) emissions from surrounding stationary sources and additionally that delta15N is more strongly correlated with surrounding stationary source NO(x) emissions than pH, SO4(2-), or NO3- concentrations. Although emission inventories indicate that vehicle emissions are the dominant NO(x) source in the eastern U.S., our results suggest that wet NO3- deposition at sites in this study is strongly associated with NO(x) emissions from stationary sources. This suggests that large areas of the landscape potentially receive atmospheric NO(y) deposition inputs in excess of what one would infer from existing monitoring data alone. Moreover, we determined that spatial patterns in delta15N values are a robust indicator of stationary NO(x) contributions to wet NO3- deposition and hence a valuable complement to existing tools for assessing relationships between NO3- deposition, regional emission inventories, and for evaluating progress toward NO(x) reduction goals.