Understanding the [NiFe] Hydrogenase Active Site Environment through Ultrafast Infrared and 2D-IR Spectroscopy of the Subsite Analogue K[CpFe(CO)(CN)2] in Polar and Protic Solvents.

The [CpFe(CO)(CN)2]- unit is an excellent structural model for the Fe(CO)(CN)2 moiety of the active site found in [NiFe] hydrogenases. Ultrafast infrared (IR) pump-probe and 2D-IR spectroscopy have been used to study K[CpFe(CO)(CN)2] (M1) in a range of protic and polar solvents and as a dry film. Measurements of anharmonicity, intermode vibrational coupling strength, vibrational relaxation time, and solvation dynamics of the CO and CN stretching modes of M1 in H2O, D2O, methanol, dimethyl sulfoxide, and acetonitrile reveal that H-bonding to the CN ligands plays an important role in defining the spectroscopic characteristics and relaxation dynamics of the Fe(CO)(CN)2 unit. Comparisons of the spectroscopic and dynamic data obtained for M1 in solution and in a dry film with those obtained for the enzyme led to the conclusion that the protein backbone forms an important part of the bimetallic active site environment via secondary coordination sphere interactions.

Masked alkynes for synthesis of threaded carbon chains.

Polyynes are chains of sp1 carbon atoms with alternating single and triple bonds. As they become longer, they evolve towards carbyne, the 1D allotrope of carbon, and they become increasingly unstable. It has been anticipated that long polyynes could be stabilized by supramolecular encapsulation, by threading them through macrocycles to form polyrotaxanes-but, until now, polyyne polyrotaxanes with many threaded macrocycles have been synthetically inaccessible. Here we show that masked alkynes, in which the C≡C triple bond is temporarily coordinated to cobalt, can be used to synthesize polyrotaxanes, up to the C68 [5]rotaxane with 34 contiguous triple bonds and four threaded macrocycles. This is the length regime at which the electronic properties of polyynes converge to those of carbyne. Cyclocarbons constitute a related family of molecular carbon allotropes, and cobalt-masked alkynes also provide a route to [3]catenanes and [5]catenanes built around cobalt complexes of cyclo[40]carbon and cyclo[80]carbon, respectively.

Optical Screening and Classification of Drug Binding to Proteins in Human Blood Serum.

Protein-drug interactions in the human bloodstream are important factors in applications ranging from drug design, where protein binding influences efficacy and dose delivery, to biomedical diagnostics, where rapid, quantitative measurements could guide optimized treatment regimes. Current measurement approaches use multistep assays, which probe the protein-bound drug fraction indirectly and do not provide fundamental structural or dynamic information about the in vivo protein-drug interaction. We demonstrate that ultrafast 2D-IR spectroscopy can overcome these issues by providing a direct, label-free optical measurement of protein-drug binding in blood serum samples. Four commonly prescribed drugs, known to bind to human serum albumin (HSA), were added to pooled human serum at physiologically relevant concentrations. In each case, spectral changes to the amide I band of the serum sample were observed, consistent with binding to HSA, but were distinct for each of the four drugs. A machine-learning-based classification of the serum samples achieved a total cross-validation prediction accuracy of 92% when differentiating serum-only samples from those with a drug present. Identification on a per-drug basis achieved correct drug identification in 75% of cases. These unique spectroscopic signatures of the drug-protein interaction thus enable the detection and differentiation of drug containing samples and give structural insight into the binding process as well as quantitative information on protein-drug binding. Using currently available instrumentation, the 2D-IR data acquisition required just 1 min and 10 µL of serum per sample, and so these results pave the way to fast, specific, and quantitative measurements of protein-drug binding in vivo with potentially invaluable applications for the development of novel therapies and personalized medicine.

Unraveling Excited State Dynamics of a Single-Stranded DNA-Assembled Conjugated Polyelectrolyte.

Conformational templating of conjugated polyelectrolytes with single-stranded DNAs (ssDNAs) has the prospect of tailoring excited state dynamics for specific optoelectronic applications. We use ultrafast time-resolved infrared spectroscopy to study the photophysics of a cationic polythiophene assembled with different ssDNAs, inducing distinct conformations (flexible disordered structures vs more rigid complexes with increased backbone planarity). Intrachain polarons are always produced upon selective excitation of the polymer, the extent being dependent on backbone torsional order. Polaron formation and decay were monitored through evolution of IR-active vibrational modes that interfere with mid-IR polaron electronic absorption giving rise to Fano-antiresonances. Selective UV excitation of ssDNAs revealed that stacking interactions between thiophene rings and nucleic acid bases can promote the formation of an intermolecular charge transfer complex. The findings inform designers of functional conjugated polymers by identifying that involvement of the scaffold in the photophysics needs to be considered when developing such structures for optoelectronic applications.

Modulation of IL-17 backbone dynamics reduces receptor affinity and reveals a new inhibitory mechanism.

Knowledge of protein dynamics is fundamental to the understanding of biological processes, with NMR and 2D-IR spectroscopy being two of the principal methods for studying protein dynamics. Here, we combine these two methods to gain a new understanding of the complex mechanism of a cytokine:receptor interaction. The dynamic nature of many cytokines is now being recognised as a key property in the signalling mechanism. Interleukin-17s (IL-17) are proinflammatory cytokines which, if unregulated, are associated with serious autoimmune diseases such as psoriasis, and although there are several therapeutics on the market for these conditions, small molecule therapeutics remain elusive. Previous studies, exploiting crystallographic methods alone, have been unable to explain the dramatic differences in affinity observed between IL-17 dimers and their receptors, suggesting there are factors that cannot be fully explained by the analysis of static structures alone. Here, we show that the IL-17 family of cytokines have varying degrees of flexibility which directly correlates to their receptor affinities. Small molecule inhibitors of the cytokine:receptor interaction are usually thought to function by either causing steric clashes or structural changes. However, our results, supported by other biophysical methods, provide evidence for an alternate mechanism of inhibition, in which the small molecule rigidifies the protein, causing a reduction in receptor affinity. The results presented here indicate an induced fit model of cytokine:receptor binding, with the more flexible cytokines having a higher affinity. Our approach could be applied to other systems where the inhibition of a protein-protein interaction has proved intractable, for example due to the flat, featureless nature of the interface. Targeting allosteric sites which modulate protein dynamics, opens up new avenues for novel therapeutic development.

Temperature-Induced Effects on the Structure of Gramicidin S.

We report on the structure of Gramicidin S (GS) in a model membrane mimetic environment represented by the amphipathic solvent 1-octanol using one-dimensional (1D) and two-dimensional (2D) IR spectroscopy. To explore potential structural changes of GS, we also performed a series of spectroscopic measurements at differing temperatures. By analyzing the amide I band and using 2D-IR spectral changes, results could be associated to the disruption of aggregates/oligomers, as well as structural and conformational changes happening in the concentrated solution of GS. The ability of 2D-IR to enable differentiation in melting transitions of oligomerized GS structures is attributed to the sensitivity of the technique to vibrational coupling. Two melting transition temperatures were identified; at Tm1 in the range 41-47 °C where the GS aggregates/oligomers disassemble and at Tm2 = 57 ± 2 °C where there is significant change involving GS ß-sheet-type hydrogen bonds, whereby it is proposed that there is loss of interpeptide hydrogen bonds and we are left with mainly intrapeptide ß-sheet and ß-turn hydrogen bonds of the smaller oligomers. Further analysis with quantum mechanical/molecular mechanics (QM/MM) simulations and second derivative results highlighted the participation of active GS side chains. Ultimately, this work contributes toward understanding the GS structure and the formulation of GS analogues with improved bioactivity.

A review of the effect of nutrient and energy restriction during late gestation on beef cattle offspring growth and development.

Changes in the environment, including nutritional changes, can influence fetal and postnatal development of the offspring, which can result in differences in growth, metabolism, reproduction, and health later in life. In beef cattle research on energy and protein restriction during late gestation appears to be contradictory. Therefore, in this review, we will examine the nutrient requirements recommended for this period. We are summarizing contradictory data on effects on offspring performance with possible explanations of the reason for why the data seems contradictory. We will finish by discussing some areas that we consider important for further research to increase the knowledge on how maternal nutrition influences offspring development. In particular, suggestions are provided on the need for more accurately measuring nutrient and energy supply and use and the impact on subsequent epigenetic effects. This will improve understanding of nutritional effects during gestation on offspring performance.

The objective of this review is to summarize the current knowledge on the effect of protein and energy restriction during the last third of gestation to beef cows on growth performance of offspring, focusing principally on Bos taurus cows. We divided the review in three sections: 1) evaluating the current beef cow requirements, highlighting areas in need of a more current research, 2) demonstrating some inconsistency on growth and carcass characteristics variables due to energy and protein restriction in late gestation, and 3) offering our opinion on future research in the area.

Environmental stress during the last trimester of gestation in pregnant cows and its effect on offspring growth performance and response to glucose and adrenocorticotropic hormone.

Winter and spring precipitation are predicted to increase in the Midwest region of the United States, causing muddy conditions. In a previous experiment, Angus cows (8 per treatment) were paired based on initial body weight (BW) and one cow from each pair was randomly allocated to either the mud or control treatment. Though cows consumed the same amount of dry matter, cows in the mud treatment weighed 37.4 kg less than cows in the control treatment by day 269 of gestation. The objective of this experiment was to evaluate developmental programming effects of steers born to cows in the mud treatment (MUD; n = 7) or the control treatment (CON; n = 6). Steers were weighed at birth and then weekly from approximately 56 d of age until weaning and were subjected to a glucose tolerance test (GTT) and adrenocorticotropic hormone (ACTH) challenge after weaning. Steers were then placed in the feedlot for an 84-d growing phase and were weighed weekly and 12th rib back fat (BF) and ribeye area (REA) were imaged every 28 d using ultrasonography. Data were analyzed as a randomized complete block design with repeated measurements when appropriate (SAS 9.4). Although there was a 37.4 kg decrease in BW of cows by the end of gestation, there was no evidence of a pen treatment effect on calf birth weight (P = 0.60) or weaning weight (P = 0.99). Additionally, there was no evidence of a pen treatment × day effect for steer BW from birth to weaning (P = 0.67) or growing phase BW (P = 0.60). There was evidence of a treatment × day of growing phase effect (P = 0.02) for BF, such that CON steers had greater BF on day 28 of the growing phase; however, there was no evidence of a treatment × day effect for REA (P = 0.20). Furthermore, there was no evidence of a pen treatment effect for the growing phase average daily gain (P = 0.74), dry matter intake (P = 0.65), gain:feed (P = 0.48), plasma glucose concentration (P = 0.67) or plasma insulin concentration (P = 0.61) in response to the GTT, or plasma cortisol concentration in response to the ACTH challenge (P = 0.51). These results indicate that while mud increased net energy requirements for cows in the MUD treatment, there were no subsequent effects observed for steer BW, gain:feed, or response to glucose and ACTH during the growing phase.

Predictions for the Midwest U.S. indicate that both winter and spring temperatures and precipitation will increase. These climatic changes could result in muddier conditions during winter and spring in the Midwest. It has previously been demonstrated that a muddy environment increases the net energy requirements of mature cow's by approximately 3.9 Mcal/d. If cows are not provided this extra energy and are exposed to mud during late gestation, it is likely that this will cause an adverse environment for the conceptus that could result in compromised growth and metabolism later in life. This study evaluated the developmental programming effects after birth and into the growing phase with beef steers born to cows that were housed in a muddy environment during late gestation compared with steers born to cows that were housed in pens bedded with wood chips during late gestation. Based on the present results, the mature cows housed in the muddy conditions weighed approximately 37.4 kg less than cows housed in pens bedded with wood chips during late gestation, however, calf birth weight, postnatal growth, postnatal feed intake and gain:feed, and postnatal response to glucose and adrenocorticotropic hormone was not affected. This indicates that the mature cows were able to mobilize body stores and supplied the fetus with adequate nutrients during gestation without impairment of growth or postnatal response to glucose and adrenocorticotropic hormone.

Chemical Markers of Human Tendon Health Identified Using Raman Spectroscopy: Potential for In Vivo Assessment.

The purpose of this study is to determine whether age-related changes to tendon matrix molecules can be detected using Raman spectroscopy. Raman spectra were collected from human Achilles (n = 8) and tibialis anterior (n = 8) tendon tissue excised from young (17 ± 3 years) and old (72 ± 7 years) age groups. Normalised Raman spectra underwent principal component analysis (PCA), to objectively identify differences between age groups and tendon types. Certain Raman band intensities were correlated with levels of advanced glycation end-product (AGE) collagen crosslinks, quantified using conventional destructive biochemistry techniques. Achilles and tibialis anterior tendons in the old age group demonstrated significantly higher overall Raman intensities and fluorescence levels compared to young tendons. PCA was able to distinguish young and old age groups and different tendon types. Raman intensities differed significantly for several bands, including those previously associated with AGE crosslinks, where a significant positive correlation with biochemical measures was demonstrated. Differences in Raman spectra between old and young tendon tissue and correlation with AGE crosslinks provides the basis for quantifying age-related chemical modifications to tendon matrix molecules in intact tissue. Our results suggest that Raman spectroscopy may provide a powerful tool to assess tendon health and vitality in the future.

Measuring proteins in H2O using 2D-IR spectroscopy: pre-processing steps and applications toward a protein library.

The ability of two-dimensional infrared (2D-IR) spectroscopy to measure the amide I band of proteins in H2O rather than D2O-based solvents by evading the interfering water signals has enabled in vivo studies of proteins under physiological conditions and in biofluids. Future exploitation of 2D-IR in analytical settings, from diagnostics to protein screening, will, however, require comparisons between multiple datasets, necessitating control of data collection protocols to minimize measurement-to-measurement inconsistencies. Inspired by analytical spectroscopy applications in other disciplines, we describe a workflow for pre-processing 2D-IR data that aims to simplify spectral cross-comparisons. Our approach exploits the thermal water signal that is collected simultaneously with, but is temporally separated from the amide I response to guide custom baseline correction and spectral normalization strategies before combining them with Principal Component noise reduction tools. Case studies show that application of elements of the pre-processing workflow to previously published data enables improvements in quantification accuracy and detection limits. We subsequently apply the complete workflow in a new pilot study, testing the ability of a prototype library of 2D-IR spectra to quantify the four major protein constituents of blood serum in a single, label-free measurement. These advances show progress toward the robust data handling strategies that will be necessary for future applications of 2D-IR to pharmaceutical or biomedical problems.

Ultrafast 2D-IR spectroscopy of [NiFe] hydrogenase from E. coli reveals the role of the protein scaffold in controlling the active site environment.

Ultrafast two-dimensional infrared (2D-IR) spectroscopy of Escherichia coli Hyd-1 (EcHyd-1) reveals the structural and dynamic influence of the protein scaffold on the Fe(CO)(CN)2 unit of the active site. Measurements on as-isolated EcHyd-1 probed a mixture of active site states including two, which we assign to Nir-SI/II, that have not been previously observed in the E. coli enzyme. Explicit assignment of carbonyl (CO) and cyanide (CN) stretching bands to each state is enabled by 2D-IR. Energies of vibrational levels up to and including two-quantum vibrationally excited states of the CO and CN modes have been determined along with the associated vibrational relaxation dynamics. The carbonyl stretching mode potential is well described by a Morse function and couples weakly to the cyanide stretching vibrations. In contrast, the two CN stretching modes exhibit extremely strong coupling, leading to the observation of formally forbidden vibrational transitions in the 2D-IR spectra. We show that the vibrational relaxation times and structural dynamics of the CO and CN ligand stretching modes of the enzyme active site differ markedly from those of a model compound K[CpFe(CO)(CN)2] in aqueous solution and conclude that the protein scaffold creates a unique biomolecular environment for the NiFe site that cannot be represented by analogy to simple models of solvation.

Open Reduction Internal fixation of Incarcerated Greater Tuberosity Avulsion Fracture: A Case Report.

Introduction: Proximal humerus fractures are common injuries of the upper extremity. These fractures are frequent in the elderly population due to low-energy trauma and in the younger patient secondary to high-energy trauma that is associated with shoulder dislocations. Proximal humerus fractures are frequently classified according to the Neer classification which is defined by the number of segments and the amount displacement required to define the fracture as displaced. Segments include the greater tuberosity (GT), the lesser tuberosity, the humeral head, and the humeral shaft. Fractures are considered displaced if there is at least 45° of angulation or 1 cm of displacement. Case Report: We present a case of a 42-year-old Caucasian patient with a displaced and incarcerated GT avulsion fracture following a motorcycle accident. In this report, we describe the patient positioning, technique using open reduction internal fixation utilizing multiple anchors and suture to treat this injury and the successful outcome of our patient 1 year from the injury. Conclusion: While standard open reduction internal fixation techniques using plates and screws are appropriate for treating these injuries, we chose to pursue a method that involved only bio-absorbable suture anchors. Using these anchors, we were able to achieve an anatomic reduction of the GT fragment that ultimately went on to achieve full healing.

Energy restriction and housing of pregnant beef heifers in mud decreases body weight and conceptus free live weight.

Average temperatures in the Midwest, USA are predicted to increase 2-9°C by the end of the century; resulting in muddy pastures for spring calving beef heifers as they enter late gestation. The objective of this study was to evaluate the effects of muddy conditions on heifer body weight (BW), body condition score (BCS), conceptus free live weight (CFLW), and fetal growth when heifers were energy restricted during late gestation. Eighteen Angus heifers (n = 9/treatment) were paired based on initial BW. One heifer from each BW pair was randomly allocated to either the mud (MUD) or control (CON) treatment on day 196 of gestation. Pens in the CON treatment were bedded with wood chips, while pens in the MUD treatment were filled with mud (average depth of 19.5 ±â€…7.9 cm). Heifers were housed individually and fed the same diet that consisted of a limit-fed total mixed ration from day 196 to 266 of gestation that was formulated to meet 66% of the net energy for maintenance, growth, and gestation requirements. Requirements and the amount of the diet offered were adjusted weekly, and heifers were weighed and sampled for blood metabolites weekly. Data were analyzed as a randomized complete block design with repeated measurements. There was a treatment × day of gestation interaction, such that heifers had similar BW, BCS, and CFLW on day 196 of gestation. By day 266 of gestation; however, heifers in the MUD treatment weighed 43.5 kg less (P < 0.01) and were 1.8 BCS units less (P < 0.01) than heifers in the CON treatment. This is further supported by the treatment × day effects we observed for back fat (BF) and rump fat (RF) thickness, such that the MUD heifers had less BF (P = 0.02) and RF (P < 0.01) by day 266 of gestation. There was a marginally significant difference for gestation length (P = 0.06), such that heifers in the MUD treatment calved approximately 3.1 days before the heifers in the CON treatment. Though heifers in the MUD treatment decreased their BW and CFLW during the treatment period, we did not observe a difference in calf birth weight (P = 0.34), calf plasma IgG concentration (P = 0.37), or calf weaning weight (P = 0.63). Despite heifers in the MUD treatment having greater BW, CFLW, and BCS losses compared with the heifers in the CON treatment, the heifers in the MUD treatment seemed to prioritize fetal growth, as they mobilized their body tissues to meet the energetic demands of pregnancy.

Effects of calcium salts of palm oil inclusion and ad libitum feeding regimen on growth performance, carcass characteristics, and plasma glucose-dependent insulinotropic polypeptide concentration of feedlot steers.

Sixty Angus × SimAngus-crossbred steers (body weight [BW] 279 ± 16 kg) were used to evaluate the effect of calcium salts of palm oil inclusion (CPO) and the amount of feed offered (AFO) on plasma glucose-dependent insulinotropic polypeptide (GIP) concentration and its association with energy metabolism and marbling score (MS) in feedlot steers. Steers were blocked by BW and gain to feed (G:F) and randomly assigned to individual feedlot pens. Treatments (2 × 2 factorial) consisted of ad libitum-fed steers without (ANF) or with (AWF) the inclusion of CPO or restricted-fed steers (85% of the ad libitum intake of ANF) without (RNF) or with the inclusion of CPO (RWF). After weaning, steers were adapted to individual pens and fed a corn silage-based diet for 30 d and subsequently placed in a ground corn (GC)-based diet. Diets were given ad libitum or at 85% of the ANF intake and with or without CPO. After 59 d on the finishing diet, all steers had ad libitum access to the finishing diet until harvest. Measurements of CO2 emission and O2 consumption to estimate respiratory quotient (RQ) were taken (n = 9/treatment). Correlations between plasma GIP and insulin concentrations and RQ were analyzed. A linear regression was performed to evaluate the association of plasma GIP and MS. All data were analyzed using the PROC MIXED procedure of SAS. During the first 103 d of the trial, there were AFO × CPO interactions (P ≤ 0.01) for BW, dry matter intake (DMI), average daily gain (ADG), and net energy for maintenance (NEm) intake. Ad libitum-fed steers without CPO presented the greatest DMI among dietary treatments and had greater BW and ADG compared with steers in the RWF and RNF treatments. After all steers had ad libitum access to dietary treatments, steers that were previously restricted showed a 30% and 19% increase (P ≤ 0.01) in ADG and G:F, respectively. There was a three-way interaction time × CPO × AFO (P = 0.04) for plasma GIP concentration. There was no correlation (P = 0.96) of GIP with RQ, whereas insulin demonstrated marginal significance for a positive (P = 0.07) and negative (P = 0.08) correlation with plasma GIP and RQ, respectively. There was no association (P = 0.30) between GIP and MS. These data indicate that GIP secretion results from an interaction between CPO and energy intake depending on the time relative to feed intake that GIP might indirectly regulate energy metabolism through insulin secretion, and that GIP does not appear to be associated with MS.

In ruminants, the role of glucose-dependent insulinotropic polypeptide (GIP) in energy metabolism has not been widely studied. This experiment is aimed to evaluate the effect of lipids, as calcium salts of palm oil (CPO), and amount of feed offered on plasma GIP concentration and its association with the energy metabolism, measured as respiratory quotient (RQ), and marbling score (MS) in feedlot steers. Steers were fed a corn silage-based diet for 30 d and subsequently placed in a ground corn-based diet until harvest. Diets were offered at different amounts (ad libitum or restricted) with or without the inclusion of CPO. Growth performance, blood samples, and gas exchange were measured over time. Overall, steers fed diets with CPO gained more body weight when they were restrict-fed but not when they were ad libitum fed. There was no association between GIP and RQ. Insulin tended to be positively and negatively correlated with plasma GIP and RQ, respectively. There was no association between GIP and MS. Our results suggest that GIP secretion results from an interaction between dietary CPO and energy intake depending on the time relative to feeding and that GIP is not associated with MS.

Short communication: A comparison between two glucose measurement methods in beef steers during a glucose tolerance test.

Glucose tolerance tests (GTT) are commonly performed in beef cattle to evaluate the glucose-insulin signaling pathway. Blood samples are obtained via a catheter and then transferred back to the laboratory for further analysis. A hand-held glucometer used chute-side can make performing GTT's and quantifying blood glucose concentration much easier and faster for research purposes. The purpose of this study was to evaluate the agreement between a hand-held electronic glucometer (Precision Xtra; Abbott Diabetes Care Inc., Mississauga, ON, Canada) for chute-side use in beef cattle compared with a colorimetric assay in the laboratory (Stanbio Glucose LiquiColor; Stanbio Laboratory, Boerne, TX, USA). A GTT was performed on 13 Simmental × Angus steers during the growing phase. Blood samples were obtained via a jugular catheter. Glucometer readings were taken immediately after blood was sampled from the jugular with no preservative, and laboratory measurements were conducted on plasma preserved with sodium fluoride. A paired t-test (P = 0.40), Pearson's correlation (P < 0.001; r = 0.95), Bland-Altman plot, and Lin's concordance correlation coefficient (LCCC = 0.90) were completed to evaluate the performance of the glucometer relative to the results from the laboratory assay. Based on the results, we conclude that the glucometer is an acceptable method for measuring blood glucose concentration in beef cattle under field conditions.

Detection of paracetamol binding to albumin in blood serum using 2D-IR spectroscopy.

Binding of drugs to blood serum proteins can influence both therapeutic efficacy and toxicity. The ability to measure the concentrations of protein-bound drug molecules quickly and with limited sample preparation could therefore have considerable benefits in biomedical and pharmaceutical applications. Vibrational spectroscopies provide data quickly but are hampered by complex, overlapping protein amide I band profiles and water absorption. Here, we show that two-dimensional infrared (2D-IR) spectroscopy can achieve rapid detection and quantification of paracetamol binding to serum albumin in blood serum at physiologically-relevant levels with no additional sample processing. By measuring changes to the amide I band of serum albumin caused by structural and dynamic impacts of paracetamol binding we show that drug concentrations as low as 7 µM can be detected and that the availability of albumin for paracetamol binding is less than 20% in serum samples, allowing identification of paracetamol levels consistent with a patient overdose.

Beef cows housed in mud during late gestation have greater net energy requirements compared with cows housed on wood chip bedding.

Mud increases net energy requirements for cattle because mud and precipitation compromise the ability of the hair coat to insulate and maintain core body temperature of the cow. The increase in energy required for a gestating cow to compensate for a muddy environment is unknown. The objective of this study was to evaluate effects of muddy conditions on cow body weight (BW) and fetal growth during late gestation. Sixteen multiparous Angus cows (n = 8/treatment) were paired based on initial BW and one cow from each pair was randomly allocated to either the mud (MUD) or control (CON) treatment on day 213 of gestation. Pens in the CON group were bedded with wood chips, while pens in the MUD group were designed to create a muddy lot (average depth of 23.6 ± 5.8 cm). Cows were housed outdoors individually and fed the same diet that consisted of a limit-fed total mixed ration. Each pair was fed to meet energy and protein requirements for maintenance and gestation. From day 213 to 269 of gestation, cows were weighed and sampled for blood metabolites weekly. Data were analyzed as a randomized complete block design with repeated measurements (SAS 9.4). Though cows consumed the same amount of dry matter, cows in the MUD treatment weighed 37.4 kg less than cows in the CON treatment (P < 0.01) by day 269 of gestation. Cows in the MUD treatment decreased approximately half a body condition score (BCS), while cows in the CON treatment gained approximately 1 BCS during the treatment period (P < 0.01). There was no evidence of a treatment × day of gestation effect for 12th rib back fat (P = 0.85), rump fat (P = 0.48), total plasma protein concentrations (P = 0.85), or plasma 3-methylhistidine (P = 0.84); however, there was a marginally significant treatment × day of gestation effect for plasma non-esterified fatty acid concentration (P = 0.09). Despite differences in cow BW at the end of the treatment period, calf birth weight (P = 0.66) and calf total plasma protein (P = 0.27) were not different; however, the divergence in cow BW remained marginally significant at parturition (P = 0.06). These results indicate that mud increased net energy requirements for cows in the MUD treatment, as calf birth weight was not different but maternal BW was decreased compared with cows in the CON treatment.

Maternal supply of a source of omega-3 fatty acids and methionine during late gestation on the offspring's growth, metabolism, carcass characteristic, and liver's mRNA expression in sheep.

The objective of the present experiment was to evaluate the effect of maternal supplementation with fatty acids (FAs) and methionine (Met) during late gestation on offspring growth, energy metabolism, plasma resolvin (RvD1) concentration, carcass characteristics, and hepatic mRNA expression. Ewes (5 pens/treatment; 3 ewes/pen) blocked by body weight (BW) were assigned to one of four treatments from day 100 of gestation until lambing. The treatments were: basal diet (NS) without FAs or Met supplementation; FA supplementation (FS; 1.01 % of Ca salts, containing n-3 FA); Met supplementation (MS; 0.1 % of rumen-protected methionine); and FS and MS (FS-MS). At birth (day 0), ewes and lambs were placed in a common pen. On day 60, lambs were weaned, sorted by sex, blocked by BW, and placed on a common finishing diet for 54 d (FP). A lamb per pen was used for a glucose tolerance test (GTT) after the FP. Carcass characteristics were recorded on day 56. Lamb data were analyzed as a randomized complete block design with a 2 × 2 × 2 factorial arrangement, with repeated measurements when needed (SAS 9.4). At weaning, lambs born to MS- or FS-fed ewes were heavier than lambs born from FS-MS ewes (FS × MS × Time; P = 0.02). A marginal significant FS × MS interaction (P = 0.09) was also observed on RvD1; lambs born to ewes in the NS and FS-MS treatments showed a lower RvD1 plasma concentration when compared with lambs born to FS- or MS-fed ewes. Lambs born to dams fed FA showed an increase (P = 0.05) in liver COX-2 mRNA relative expression. Lambs born to ewes supplemented with Met showed an increase (P = 0.03) in liver FABP4 mRNA expression. An FS × MS × Time interaction (P = 0.07) was observed in plasma glucose during the GTT; lambs born from FS-fed ewes showed lower plasma glucose concentration than lambs born to Met-supplemented ewes at 2 min after bolus administration. During the GTT, a marginal significant effect (P = 0.06) was observed for the lamb average insulin concentration due to maternal Met supplementation during late gestation, where these lambs had the lowest plasma concentration. Contrary to our hypothesis, the interaction of FA and Met supplementation during late gestation did not show a greater positive effect on offspring postnatal growth and metabolism. However, the individual supplementation of each nutrient has an effect on offspring development with a concomitant change in markers involved in the inflammatory response and energy metabolism.

Late gestation supplementation of omega-3 fatty acids (FAs) or methionine (Met) alters the offspring's development. However, the effect of both nutrients on the physiology and growth of the progeny has not been explored. The experiment's objective was to evaluate the effect of dam supplementation with a Met and the omega-3 FAs, such as eicosapentaenoic and docosahexaenoic acids, during late gestation on growth, energy metabolism, and inflammatory response markers of the lamb. Ewes received one of the four following treatments: basal diet without FA or Met, FA supplementation, Met supplementation, or FA and Met supplementation. Supplementation of omega-3 FAs and Met did not show a greater effect on postnatal growth and metabolism of the offspring compared with the supplementation of each nutrient individually. However, individual supplementation influences offspring development with a concomitant change in markers involved in the inflammatory response and energy metabolism. Results of the present experiment suggest that offspring born to mothers that were fed FAs or Met on late gestation could have the ability to better cope with inflammatory processes, which could improve their long-term growth performance. Moreover, maternal supplementation of Met during late gestation could modulate offspring's glucose­insulin system, which may also affect offspring's growth.

Digital storytelling within the Australian mining industry: worker engagement and health literacy indicator effects.

The mining industry is a demanding context for workplace health education due to a range of factors including productivity targets, workforce diversity and work roster schedules. This project investigated the impact of digital story health communication on worker engagement and its effect on interactive and critical health literacy indicators. The study comprised a quasi-experimental parallel time series research design, with control and intervention groups at each of the mine sites (n = 2). Workers in the intervention group (n = 85) received a 'toolbox talk' presentation incorporating a digital story featuring a mining industry worker and a leading cardiovascular health expert. The control group (n = 90) received equivalent health information communicated in a non-narrative manner, reflective of typical practices within the mining industry. A significantly greater effect was evident for worker engagement within the intervention group, with substantial maintenance over the follow-up period, compared with no significant effect at follow-up within the control group. Significant effects on interactive health literacy indicators (n = 3) were evident for the intervention group with corresponding lower level or nil effects within the control group. The findings highlight the benefits of evidence-based digital stories as an efficient and efficacious worker-centred health communication strategy for complex industrial workplace environments.

This quasi-experimental intervention study sought to test the effectiveness of digital stories featuring mining workers and leading health experts as a workplace health education strategy. The mining industry is a challenging setting for health education due to a range of reasons including productivity targets, workforce diversity and work roster schedules. Workers in the intervention group received a 'toolbox talk' presentation including a digital story. Workers in the control group received the same health information without the digital story, reflecting common health communication practices in the industry. Outcomes of this research demonstrate that digital storytelling was more effective on worker engagement and interactive health literacy, with a promising level of maintenance over a follow-up period. The findings show the benefits of digital storytelling as a worker focused health education strategy for complex industrial settings where time efficient communication methods are required.

Social Network Differences Between American Indian Youth Who have Attempted Suicide and Have Suicide Ideation.

Suicide is a crucial public health concern for American Indian and Alaska native (AIAN) communities. AIANs have the highest suicide rate compared to all other ethnic groups in the United States. Social relations are a salient fixture of AIAN culture. The primary aims of this study were to describe the personal networks of AI youth that have recently had a suicide attempt or suicidal ideation and to identify key network differences between the two groups. This study uses personal networks collected among AIs living on a reservation in the Southwest. Our sample included 46 American Indians that have recently attempted suicide or had suicidal ideation. We explored social network characteristics of the two groups descriptively as well as comparatively (t-tests). Our findings suggest that AI youth that have attempted suicide nominate more friends in their networks that have used alcohol and drugs compared to the networks of AI youth that have recent suicide ideation. Additionally, AI youth that recently attempted suicide have used alcohol and drugs with their network peers at a higher rate than youth that have had recent suicide ideation. Lastly, AI youth that have attempted suicide recently were significantly more likely to have more nominated friends in their networks that they had reached out to when they were struggling with suicide compared to their peers that have experienced recent suicide ideation. These results indicate a promising method moving forward to identify unique intervention strategies that extend beyond the individual.