MIXED Procedure Research Articles

Horses with previous episodes of laminitis have altered insulin responses to seasonal oral sugar testing and grazing compared to horses with no known history of laminitis

Forage is essential for equine health and performance, but intake of elevated pasture nonstructural carbohydrates (NSC) may exacerbate metabolic disorders. This study aimed to investigate the influence of laminitis history on metabolic and morphometric responses in grazing horses. Twelve non-pregnant mares (15 ± 3.4 yrs) were selected based on previous diagnosis of laminitis (PRELAM; n=6) or not (NOLAM; n=6). Horses were maintained on 8.5-ha pasture. Weekly pasture samples were clipped at random for nutrient analysis. Monthly blood samples were collected and analyzed for glucose and insulin concentrations. Body weight (BW), body condition score (BCS), and cresty neck score (CNS) were evaluated monthly. Each month, a modified oral sugar test (OST) was used to assess basal (T0) and 75-minute (T75) insulin dynamics following 0.30 mL/kg BW Karo Light Corn Syrup. Data were analyzed using PROC MIXED with repeated measures (SAS Institute). PRELAM had greater CNS (P < 0.001), BW (P < 0.05), and BCS compared to NOLAM (P < 0.05). Non-fasted glucose was influenced by month (P < 0.001) while non-fasted insulin was affected by group by month (P = 0.004). Fasted insulin concentrations (T0) were affected by month and group (P < 0.001) where PRELAM had greater concentrations. T75 glucose and insulin concentrations were affected by month (P < 0.001 and P = 0.002, respectively) and were higher for PRELAM (P = 0.003). These data suggest horses with greater general and regional adiposity, and previous history of laminitis, often exhibit greater insulin concentrations on pasture and to OST.

Effects of cashew nutshell extract inclusion into a high-grain finishing diet on methane emissions, nutrient digestibility, and ruminal fermentation in beef steers.

By 2050, the U.S. beef industry must produce an extra 40 million tons of beef to satisfy the global demand. Such an increase in inventory will undoubtedly enhance methane (CH4) production from livestock, which should be reduced by over 20%. The addition of plant secondary metabolites, such as anacardic acid present in cashew nutshell extract (CNSE), has shown promising results in reducing CH4 yield, although its effects seemed to be diet dependent. This study evaluated the addition of CNSE to a high-grain diet (85:15 Grain: forage) on in vivo CH4 emissions, nutrients digestibility, performance, feeding behavior, and ruminal fermentation parameters of beef steers. Sixteen Angus crossbred steers [599 ± 40kg of bodyweight (BW)] and six ruminally cannulated crossbred steers (490 ± 51kg of BW) were utilized in a crossover design with 2 experimental periods of 56 d each, composed by 14 d of adaptation, 35 d of measurement, and 7 d of washout. Following adaptation, steers were sorted by BW, and assigned to receive no additive (CON) or CNSE at 5g/steer/d. Data were analyzed using the MIXED procedure of SAS. Inclusion of CNSE increased (P < 0.05) propionate concentration and molar proportion (MP; mol/100mol), tended to decrease acetate MP (P = 0.10), reduced the acetate: propionate (A:P) ratio (P = 0.05), and MP of branched chain volatile fatty acids (P < 0.01). Neither in vitro organic matter digestibility nor in vitro CH4 yield were affected by CNSE inclusion (P > 0.05). Steers receiving CNSE exhibited greater (P < 0.05) final BW, dry matter intake (DMI), and average daily gain (ADG) but lesser (P < 0.05) in vivo CH4 emission rate (g/d), yield (g/kg of DMI), and intensity (g/kg of ADG). Meal length, bunk visit duration, and apparent total tract digestibility of DM increased (P < 0.05) after CNSE addition. Considering CNSE-supplemented steers spent more time in the feedbunk and exhibited higher DMI, CH4 mitigation was unlikely associated with intake reduction. The addition of CNSE to a high-grain diet in beef steers demonstrated significant improvements in animal performance and reduced CH4 emissions, as the result of shifts in ruminal fermentation patterns, favoring propionate instead acetate concentration, leading to a reduction in the A:P ratio. CNSE shows promise as a strategy to enhance beef industry sustainability.

Comparison Between Conventional and Artificial Intelligence-Assisted Setup for Digital Implant Planning: Accuracy, Time-Efficiency, and User Experience.

To investigate the reliability and time efficiency of the conventional compared to the automatic artificial intelligence (AI) segmentation of the mandibular canal and registration of the CBCT with the model scan data, in relation to clinician's experience. Twenty clinicians, 10 with a moderate and 10 with a high experience in computer-assisted implant planning, were asked to perform a bilateral localization of the mandibular canal, followed by a registration of the intraoral model scan with the CBCT. Subsequently, for each data set and each participant, the same operations were performed utilizing the AI tool. Statistical significance was assessed via a mixed model (using the PROC MIXED statement and the compound symmetry covariance structure). The mean time for the segmentation of the mandibular canals and the registration of the models was 4.75 (2.03)min for the manual and 2.03 (0.36) min for the AI-automated operations (p < 0.001). The mean discrepancy in the mandibular canals was 0.71 (1.80) mm RMS error for the manual segmentation and 0.68 (0.36) RMS error for the AI-assisted segmentation (p > 0.05). For the registration between the CBCT and the intraoral scans, the mean discrepancy was 0.45 (0.16) mm for the manual and 0.37 (0.07) mm for the AI-assisted superimposition (p > 0.05). AI-automated implant planning tools are feasible options that can lead to a similar or better accuracy compared to the conventional manual workflow, providing improved time efficiency for both experienced and less experienced users. Further research including a variety of software and data sets is required to be able to generalize the outcomes of the present study.

Effects of cutting schedule and inoculation on chemical composition, ensiling characteristics and digestive utilisation of alfalfa silages

ABSTRACT The objective of this study was to evaluate the effects of ensiling alfalfa cut in the morning or the afternoon with or without inoculant on chemical composition, ensiling characteristics, and digestive utilisation of alfalfa silages. Fresh forage was obtained from 3 plots of alfalfa, each one subdivided into 12 subplots. The experimental design was a 2 × 2 factorial (cut AM or PM and with or without inoculant), with three replications (alfalfa plots) and three subsamples (subplots). Forage from each subplot was employed to fill one experimental silo. Variables of chemical composition, fermentation characteristics, and in vitro digestive utilisation were determined. Data obtained were analysed by a mixed procedure and least square means were compared. Correlations between variables of chemical composition of original forage and composition and fermentation of silages, as well as between variables of chemical composition and digestive utilisation of silages were analysed. PM cutting led to silages with higher content of remaining soluble sugars (P = 0.01), lower pH (P < 0.01), lower NH3–N concentration (P = 0.02), and higher IVTDMD (P = 0.03) than silages made in the morning. Providing enough WSC through cutting forage in the afternoon led to improvements in conservation quality and in vitro digestive utilisation and was more effective than inoculating alfalfa.

A pre- and postnatal immune challenge influences muscle growth and metabolism in weaned pigs.

The in-utero environment is key to both fetal and postnatal growth and development. The objective of this study was to determine if administration of an acute low-dose lipopolysaccharide (LPS) to gestating sows during mid to late gestation and post-weaning would alter the offsprings metabolomic profile of the longissimus dorsi (LD) and muscle ultrastructure. Pregnant Camborough sows were randomly assigned to receive LPS (LPS; n= 7) at a dose of 2.5 μg/kg or saline (CON; n = 7) on 78 ± 1.8 d of gestation. At weaning (21 ± 1.3 d of age), barrows (CON n = 17; LPS n = 17) from each treatment were selected to receive a secondary LPS. Barrows were administered the secondary LPS challenge at a dose of 10 μg/kg 7 d post weaning. Twenty-four h after the postnatal LPS dose, barrows (31 ± 1.3 d of age) were euthanized, and each LD was removed. The left LD was utilized for morphometric measurements. Two samples from the medial section of the right LD were preserved for immunohistochemical measurements and metabolomic analyses. Mass spectral data were deconvoluted, aligned, and annotated using MS-DIAL. Univariate and multivariate analyses were conducted using MetaboAnalyst. Pathway analysis was conducted and compared to the Homo sapiens pathway library. Morphometric and immunohistochemical measurements were analyzed using the MIXED procedure of SAS version 9.4. Significance for all analyses was declared at P ≤ 0.05 and tendencies were considered at P ≤ 0.10. Average diameter of myosin heavy chain (MHC) type I and IIB/X fibers was increased (P ≤ 0.048) in LPS offspring compared with CON. Average cross-sectional area was increased (P = 0.030) in MHC IIB/X fibers and tended to be increased (P = 0.080) in MHC I fibers of LPS offspring. There were no differences (P ≥ 0.186) between treatment groups for total nuclei or nuclei positive for MYF5, PAX7, or MYF5 and PAX7 nuclei. Metabolomic analyses identified 14 differentially expressed (P < 0.05) metabolites in the LD between treatment groups. There were 10 metabolites within the LD that tended (P ≤ 0.096) to differ between treatment groups. Thus, this study shows that in-utero immune stimulation using LPS in gestating sows and a subsequent LPS challenge postnatally alters the metabolomic profile and muscle ultrastructure of the LD in weaned pigs.

Rheology, shrinkage, mechanical properties and microstructure of ultra-light-weight concrete with fly ash cenospheres

Cenospheres are low-density and hollow microspheres derived from coal-fired power plant fly ash waste. This study aims to prepare ultra-light-weight (<1000 kg/m3 wet density) concrete using fly ash cenospheres (FAC). To begin with, FAC's shell thickness and the water absorption and desorption were characterized. A mixing procedure was designed to avoid the segregation between the FAC and cement slurry. FAC can affect the rheological properties of fresh mixture over time through absorption and desorption of free water. The presented ultra-light-weight concrete has several advantages compared to the ones prepared using foaming methods. First, shrinkage is significantly reduced due to FAC's internal restraint and curing effects. Secondly, it has good mechanical performance, especially in bending and is more environmentally friendly due to use of less cement. X-ray computed tomography illustrates that FAC ultra-light-weight concrete has smaller pores of more uniform size compared with those prepared using foaming methods. X-Ray diffraction, thermal gravimetry-derivative thermal gravimetry, fourier-transform infrared spectroscopy and scanning electron microscopy are employed for the hydration products and microstructure characterization. Outcomes prove that FAC can combine well with the cement matrix, and react with calcium hydroxide to produce C-A-S-H through pozzolanic reaction.

Impacts of replacing soybean meal with processed soybean meal on intestinal health and growth of nursery pigs challenged with F18+ Escherichia coli.

This study aimed to investigate the impact of different level of soybean meal (SBM) replaced by soy protein concentrate on intestinal health and growth performance of nursery pigs under F18+ Escherichia. coli (E. coli). Forty-eight newly weaned pigs (6.6 ± 0.3 kg) were randomly allotted to 4 treatments arranged by 2 × 2 factors using randomized complete block design with initial body weight and sex as blocks. Two factors were F18+ E. coli challenge (0 or 2.1 × 1010 CFU) and the level of SBM (24% or 12% in phase 1 and 26% or 14% in phase 2). Pigs were fed for 25 d in 2 phases (phase 1 for 11 d and phase 2 for 14 d). At the end of study, all pigs were euthanized to collect jejunal mucosa and tissues to measure parameters related to intestinal health. Data were analyzed using a MIXED procedure in SAS. The F18+ E. coli challenge decreased (p<0.05) overall average daily gain (ADG) and average daily feed intake (ADFI) and decreased (p<0.05) gain to feed ratio on d 7 to 11. The High SBM tended to have a greater overall ADG (p=0.054) and ADFI (p=0.078) compared with low SBM under F18+ E. coli challenge, but not in unchallenged conditions. The F18+ E. coli challenge increased (p<0.05) fecal score on d 7 to 18. The tumor necrosis factor-alpha and interleukin-1β in jejunal mucosa were decreased (p<0.05) in High SBM treatments. The High SBM tended to increase (p=0.085) occludin expression in jejunum. High SBM increased crypt depth in jejunum under F18+ E. coli challenge, but not in unchallenged conditions (p<0.05). High SBM in nursery diets could alleviate the detrimental effects of F18+ E. coli challenge on growth performance of pigs under compared to low SBM inclusion, which might be attributed to decreased intestinal inflammation and improved intestinal integrity.

Impacts of Protein and Energy Levels on Rumen Fermentation and Microbial Activity Under Different Incubation Temperatures.

This study aimed to explore the effects of different incubation temperatures on ruminal fermentation and rumen microorganisms and determine the appropriate protein and energy levels to enhance microbial protein synthesis using an in vitro system. Rumen inoculum was collected from two fistulated Holstein heifers (trial 1: BW: 652.3 kg ± 25.2; trial 2: BW: 683.3 kg ± 30.2) and assessed using a closed-batch culture system. The experimental model employed a 2 × 5 factorial arrangement using incubation temperatures set to 39 and 41 °C, with protein levels set to 12.0, 13.5, 15.0, 16.5, and 18.0% of DM in trial 1 or with energy levels set to 2.4, 2.5, 2.6, 2.7, and 2.8 Mcal/kg of DM in trial 2. The data were analyzed using the MIXED procedure. The results showed increased (p < 0.05) NH3-N concentrations and total volatile fatty acids (TVFAs) with higher incubation temperatures, while the liquid-associated bacterial (LAB) amounts decreased (p < 0.05) in trials 1 and 2. The interaction between the energy level and incubation temperature affected (p < 0.05) the LAB protein levels in trial 2. Higher protein levels led to increased (p < 0.05) NH3-N and acetate concentrations, but it decreased (p < 0.05) the propionate percentage. Conversely, higher energy levels decreased (p < 0.05) the amount of acetate and increased the propionate concentration, altering the acetate-to-propionate ratio. However, no interaction involving TVFA and LAB was observed between the incubation temperature and the protein or energy levels. Changes in the NH3-N, TVFAs, and LAB protein amounts were observed under different incubation temperatures and energy levels. In conclusion, these findings provide insight into the metabolic adaptation under different ruminal temperatures and the impacts of dietary adjustments on rumen fermentation and microbial activity. However, there are limitations to replicating the complex physiological responses that occur within the whole body solely through in vitro experiments.

Injectable progesterone in 13-month-old prepubertal Nellore heifers: effects on in vitro embryo production and pregnancy rate.

The aim of this study was to evaluate the effect of injectable progesterone administration before ovum pick-up (OPU) on in vitro embryo production (IVEP) and pregnancy rate in prepubertal Nellore heifers. Twenty-three Nellore females, 13.04 ± 0.8 months of age, were randomly assigned to two groups: control group (no progesterone application) and progesterone group (intramuscular administration of 1 mL [150mg/mL] progesterone 7 days before OPU). The second OPU was performed after 28 days (crossover between groups). After OPU, oocytes were selected for IVEP and a sample was fixed in 4% paraformaldehyde. Viable oocytes were subjected to immunofluorescence staining for IGFBP2 and caspase-3. For each oocyte, one image was generated per channel (A555 and A488 filters) and analyzed with the ImageJ program for the quantification of fluorescence intensity (in pixels). The produced embryos were transferred to multiparous Nellore cows and pregnancy was diagnosed by ultrasound 30 days after transfer. Data were analyzed using the MIXED and GLIMMIX procedure of SAS®. Treatment with injectable progesterone had no effect on the number or size of the antral follicles of heifers. No differences between the control and progesterone groups were observed for total number of aspirated oocytes (P = 0.78), viable oocytes (P = 0.80), cleavage rate (P = 0.60), or blastocyst rate (P = 0.56). There were no differences in pixel intensity between the two groups, nor in the levels of IGFBP2 or caspase-3. However, a difference was observed in the pregnancy rate of embryos produced from heifers of the progesterone group (42.1%; 32/76) when compared to the rate of embryos produced from control heifers (28.4%; 23/81) (P = 0.02). The use of injectable progesterone before OPU in prepubertal Nellore heifers does not improve IVEP but increases the pregnancy rate after transfer.

Standardized Ileal Digestibility of Amino Acids in Hybrid Rye Ground to Two Particle Sizes and Fed With or Without Multienzyme Supplement to Young Growing Pigs.

A newly developed hybrid rye with enhanced ergot resistance has potential as an alternative energy source for pigs. An experiment was conducted to evaluate the effects of particle size (PS) and multienzyme supplement (MES) on the apparent total tract digestibility (ATTD) of energy and nutrients and the standardized ileal digestibility (SID) of amino acids (AA) in hybrid rye fed to growing pigs. Bono and Gatano, two hybrid rye varieties, were ground to either a coarse (1111 and 1210 μm) or a fine (594 and 717 μm) PS using a hammer mill. Eighteen ileal-cannulated barrows (initial BW = 18.2 ± 1.0 kg) were randomly assigned to 1 of 9 dietary treatments in a replicated 9 × 3 incomplete Latin square design to give six observations per treatment. Among the nine experimental diets, eight featured two hybrid rye varieties (Bono or Gatano), either coarsely or finely ground, as the only source of AA with or without MES, while an N-free diet was used to estimate the endogenous losses of AA at the distal ileum. All diets contained titanium dioxide as an indigestible marker. Each period lasted 9 days, with the first 5 days being for adaptation followed by 2 days each for faecal and ileal digesta collection. Data were analysed using the MIXED procedure of SAS, with the final model having variety, PS, MES, their two-way interactions, and a three-way interaction. No interactions were noted except that the apparent ileal digestibility and SID of crude protein (CP) and most AA in Gatano had greater effects of MES than Bono (Variety × MES; p < 0.05). The ATTD of dry matter (DM), gross energy (GE) and CP were greater (p < 0.05) in Bono diets than in Gatano diets. Finely ground hybrid rye diets had higher (p < 0.05) ATTD of neutral detergent fibre assayed with a heat stable amylase and expressed inclusive of residual ash (NDF) and ether extract compared to coarsely ground hybrid rye diets. Dietary MES supplementation increased (p < 0.05) the ATTD of DM, GE, CP, NDF, acid detergent fibre expressed inclusive of residual ash, and ash in hybrid rye diets. In conclusion, the nutrients digestibility of hybrid rye can be affected by its variety. PS reduction improved the nutrient digestibility in hybrid rye, and MES supplementation improved the energy and AA digestibility in hybrid rye fed to growing pigs.

A Nonparametric Global Win Probability Approach to the Analysis and Sizing of Randomized Controlled Trials With Multiple Endpoints of Different Scales and Missing Data: Beyond O'Brien-Wei-Lachin.

Multiple primary endpoints are commonly used in randomized controlled trials to assess treatment effects. When the endpoints are measured on different scales, the O'Brien rank-sum test or the Wei-Lachin test for stochastic ordering may be used for hypothesis testing. However, the O'Brien-Wei-Lachin (OWL) approach is unable to handle missing data and adjust for baseline measurements. We present a nonparametric approach for data analysis that encompasses the OWL approach as a special case. Our approach is based on quantifying an endpoint-specific treatment effect using the probability that a participant in the treatment group has a better score than (or a win over) a participant in the control group. The average of the endpoint-specific win probabilities (WinPs), termed the global win probability (gWinP), is used to quantify the global treatment effect, with the null hypothesis gWinP = 0.50. Our approach involves converting the data for each endpoint to endpoint-specific win fractions, and modeling the win fractions using multivariate linear mixed models to obtain estimates of the endpoint-specific WinPs and the associated variance-covariance matrix. Focusing on confidence interval estimation for the gWinP, we derive sample size formulas for clinical trial design. Simulation results demonstrate that our approach performed well in terms of bias, interval coverage percentage, and assurance of achieving a pre-specified precision for the gWinP. Illustrative code for implementing the methods using SAS PROC RANK and PROC MIXED is provided.

Influence of Ensiling Timing and Inoculation on Whole Plant Maize Silage Fermentation and Aerobic Stability (Preliminary Research)

Despite efforts to prevent atypical ensiling conditions, such as delayed ensiling or sealing, these issues frequently occur in practice. This study aimed to investigate the effects of delayed ensiling (forage held for 24 h) and sealing, along with inoculation using a blend of Lentilactobacillus buchneri and Lactococcus lactis, on the characteristics of the resulting silages. Whole-plant maize (Zea mays L.) was treated with or without a commercial inoculant and ensiled (36% dry matter) for 60 days in 3.0 L glass containers. The forage was either ensiled immediately or subjected to a 24 h delay before ensiling. During the delay, the forage was either covered or left uncovered. Each treatment was replicated five times. All data were analyzed using the MIXED procedure of SAS statistical software (version 9.4; SAS Institute Inc., Cary, NC, USA). Delaying the ensiling process by 24 h worsens fermentation parameters, significantly increases dry matter (DM) losses (p &lt; 0.01), and significantly reduces aerobic stability and the hygienic quality of the silage (p &lt; 0.01), as evidenced by higher concentrations of undesirable fermentation products and elevated yeast and mold counts. The inoculation has a significant impact on both forage before ensiling and the characteristics of the resulting silage. Maize forage treated with inoculant showed a lower temperature increase by 8.2–8.1 °C (p &lt; 0.01) when delayed for 24 h before ensiling. In silages, it also resulted in a reduced pH (p &lt; 0.01); increased concentrations of lactic acid; acetic acid; and 1,2-propanediol (p &lt; 0.01); and decreased levels of negative fermentation indicators such as ammonia-N, alcohols, and butyric acid (p &lt; 0.01) During both the fermentation and aerobic exposure periods, inoculated silages exhibited up to 36% and 2.6 times lower (p &lt; 0.01) dry matter loss, while suppressing the growth of yeasts and molds by up to 2.6 and 3.1 times (p &lt; 0.01), respectively, compared to non-inoculated silages. The results of this study support the recommendation to minimize the duration of aerobic exposure of fresh forage during silo filling and to use LAB-based inoculants.

Seasonal environmental fluctuations alter the transcriptome dynamics of oocytes and granulosa cells in beef cows

BackgroundExamining the mechanistic cellular responses to heat stress could aid in addressing the increasing prevalence of decreased fertility due to elevated ambient temperatures. Here, we aimed to study the differential responses of oocytes and granulosa cells to thermal fluctuations due to seasonal differences. Dry beef cows (n = 10) were housed together, synchronized and subjected to a stimulation protocol to induce follicular growth before ovum pick-up (OPU). Two OPU’s were conducted (summer and winter) to collect cumulus-oocyte-complexes (COCs) and granulosa cells. In addition, rectal temperatures and circulating blood samples were collected during OPU. Oocytes were separated from the adherent cumulus cells, and granulosa cells were isolated from the collected OPU fluid. RNA was extracted from pools of oocytes and granulosa cells, followed by library preparation and RNA-sequencing. Blood samples were further processed for the isolation of plasma and leukocytes. The transcript abundance of HSP70 and HSP90 in leukocytes was evaluated using RT-qPCR, and plasma cortisol levels were evaluated by immunoassay. Environmental data were collected daily for three weeks before each OPU session. Data were analyzed using MIXED, Glimmix or GENMOD procedures of SAS, according to each variable distribution.ResultsAir temperatures (27.5 °C vs. 11.5 °C), average max air temperatures (33.7 °C vs. 16.9 °C), and temperature-humidity indexes, THI (79.16 vs. 53.39) were shown to contrast significantly comparing both the summer and winter seasons, respectively. Rectal temperatures (Summer: 39.2 ± 0.2 °C; Winter: 38.8 ± 0.2 °C) and leukocyte HSP70 transcript abundance (Summer: 4.18 ± 0.47 arbitrary units; Winter: 2.69 ± 0.66 arbitrary units) were shown to increase in the summer compared to the winter. No visual differences persisted in HSP90 transcript abundance in leukocytes and plasma cortisol concentrations during seasonal changes. Additionally, during the summer, 446 and 940 transcripts were up and downregulated in oocytes, while 1083 and 1126 transcripts were up and downregulated in the corresponding granulosa cells, respectively (Fold Change ≤ -2 or ≥ 2 and FDR ≤ 0.05). Downregulated transcripts in the oocytes were found to be involved in ECM-receptor interaction and focal adhesion pathways, while the upregulated transcripts were involved in protein digestion and absorption, ABC transporters, and oocyte meiosis pathways. Downregulated transcripts in the granulosa cells were shown to be involved in cell adhesion molecules, chemokine signaling, and cytokine-cytokine receptor interaction pathways, while those upregulated transcripts were involved in protein processing and metabolic pathways.ConclusionIn conclusion, seasonal changes dramatically alter the gene expression profiles of oocytes and granulosa cells in beef cows, which may in part explain the seasonal discrepancies in pregnancy success rates during diverging climatic weather conditions.

Effects of dietary Lactobacillus postbiotics and bacitracin on the modulation of mucosa-associated microbiota and pattern recognition receptors affecting immunocompetence of jejunal mucosa in pigs challenged with enterotoxigenic F18+ Escherichia coli

BackgroundEnterotoxigenic Escherichia coli (E. coli) is a threat to humans and animals that causes intestinal disorders. Antimicrobial resistance has urged alternatives, including Lactobacillus postbiotics, to mitigate the effects of enterotoxigenic E. coli.MethodsForty-eight newly weaned pigs were allotted to NC: no challenge/no supplement; PC: F18+ E. coli challenge/no supplement; ATB: F18+ E. coli challenge/bacitracin; and LPB: F18+ E. coli challenge/postbiotics and fed diets for 28 d. On d 7, pigs were orally inoculated with F18+ E. coli. At d 28, the mucosa-associated microbiota, immune and oxidative stress status, intestinal morphology, the gene expression of pattern recognition receptors (PRR), and intestinal barrier function were measured. Data were analyzed using the MIXED procedure in SAS 9.4.ResultsPC increased (P < 0.05) Helicobacter mastomyrinus whereas reduced (P < 0.05) Prevotella copri and P. stercorea compared to NC. The LPB increased (P < 0.05) P. stercorea and Dialister succinatiphilus compared with PC. The ATB increased (P < 0.05) Propionibacterium acnes, Corynebacterium glutamicum, and Sphingomonas pseudosanguinis compared to PC. The PC tended to reduce (P = 0.054) PGLYRP4 and increased (P < 0.05) TLR4, CD14, MDA, and crypt cell proliferation compared with NC. The ATB reduced (P < 0.05) NOD1 compared with PC. The LPB increased (P < 0.05) PGLYRP4, and interferon-γ and reduced (P < 0.05) NOD1 compared with PC. The ATB and LPB reduced (P < 0.05) TNF-α and MDA compared with PC.ConclusionsThe F18+ E. coli challenge compromised intestinal health. Bacitracin increased beneficial bacteria showing a trend towards increasing the intestinal barrier function, possibly by reducing the expression of PRR genes. Lactobacillus postbiotics enhanced the immunocompetence of nursery pigs by increasing the expression of interferon-γ and PGLYRP4, and by reducing TLR4, NOD1, and CD14.

Inclined treadmill short-duration exercise for walk regularity of recreation horses

Knowledge on horse symmetry and regularity is essential for every training/rehabilitation programs. Present studies compare biomechanical parameters for different treadmill positions, but not the effect of such exercise. The aim of the presented work was to study recreation horse movement parameters before and immediately after inclined short-duration exercise in walk. The hypothesis was that inclined short-duration exercise gave greater stride regularity and symmetry. Walk biomechanical parameters of 12 Warmblood recreation horses were monitored as temporal data using video analysis [50 frames/s] before and after 5-minute inclined work on a treadmill at 3% incline. Control group (6 horses) consisted from earlier evaluated horses that performed on the other day the same walk exercise without incline. Analysis of variance was conducted for temporal and spatial parameters of walk (237 measurements for incline and 109 for control) using the Mixed SAS procedure. For both data sets the statistical model with the random effect of horse, fixed effects of exercise and successive stride was applied. Control group showed not differences before/after short walk exercise. The influence of inclined exercise was statistically significant for the lateral limbs sequence, regularity and standard deviation of strides regularity (p<0.05). The lateral limbs sequence and regularity increased after inclined exercise (3–5%). The standard deviation of regularity was almost 2-fold lower. Inclined treadmill exercise has a positive effect on walk regularity.

Effect of scanning strategies on trueness and time efficiency of digital impression on paediatric models.

Various scanning methods aim to reduce intraoral scanner errors, yet no specific approach targets paediatric patients. Evaluate continuous (C) and noncontinuous (NC) scanning patterns' impact on the trueness and duration of paediatric digital impressions. A standard pedodontic typodont model was scanned with a Trios 4 Move+ IOS using four scanning strategies. C1 and C2 followed continuous scanning from the right molars, with different directions. NC1 scanned noncontinuously from the right molar, with breaks every four teeth. NC2 began at the right primary incisor, with breaks after every three teeth. Each scan, performed five times by one researcher, was timed. Industrial scanning provided reference data. Deviations were analyzed using the MIXED procedure. NC1 had higher deviations in the lower jaw, whereas C1 and NC1 showed higher deviations in the upper jaw (p < .001). C1 had significantly higher deviations in the upper jaw than the lower jaw (p = .041). NC2 had the longest scanning time in both upper and lower jaws (p = .002). Paediatric digital impressions benefit from starting at the anterior and incorporating breaks, although this increases scanning time.

Bone loss after bariatric surgery is observed mainly in the hip trabecular compartment and after hypoabsorptive techniques

We evaluated the impact of bariatric surgery on bone mineral density (BMD) and microarchitecture over one year using dual-energy X-ray absorptiometry (DXA), the trabecular bone score (TBS), and 3D-DXA to assess changes after different surgical techniques. This prospective, single-center study of 153 patients with severe obesity contrasts the effects on bone health of sleeve gastrectomy (SG), Roux-en-Y gastric bypass (RYGB), and duodenal switch/single anastomosis duodeno-ileostomy with sleeve gastrectomy (DS/SADIS). To our knowledge, this is the first study to evaluate patients undergoing DS/SADIS and to incorporate 3D-DXA analysis in the assessment of bone loss.Patients were 81 % female with a mean age of 50 ± 9 years. Fifty-four per cent underwent SG; 16 %, RYGB; and 30 %, DS/SADIS. Our findings revealed a significant decrease in areal BMD at the LS (−3.49 ± 5.44 %), FN (−5.24 ± 5.86 %), and TH (−8.06 ± 5.14 %) one year after bariatric surgery. Bone microarchitecture at the LS assessed by TBS was degraded in 30 % of patients. Proximal femur 3D-DXA analysis showed that surgery-induced bone loss predominantly affects the trabecular compartment (Trabecular volumetric (v) BMD: −8.00 ± 6.57 %) rather than the cortical compartment (Cortical vBMD: −1.37 ± 2.79 %).These results suggest hypoabsorptive and mixed techniques (DS/SADIS and RYGB) were associated with greater BMD loss and deterioration of microarchitecture than restrictive techniques (SG).The primary determinants of bone density and impairment of microarchitecture were the extent of weight loss and the type of surgical procedure. Despite overall bone loss, Z-score assessments indicated that post-surgical bone status remained within or above the average ranges compared to a healthy population, except for TH following DS/SADIS.In conclusion, our research shows differences in the impact of bariatric surgery techniques on bone density and microarchitecture, emphasizing the need for careful postoperative monitoring of bone health, particularly in patients undergoing hypoabsorptive and mixed procedures.

Validation of a Commercial ELISA Kit for Non-Invasive Measurement of Biologically Relevant Changes in Equine Cortisol Concentrations

The measurement of fecal cortisol/corticosterone metabolites (FCMs) is often used to quantify the stress response. The sampling method is relatively non-invasive, reduces concern for elevation of cortisol from the sampling method, and has been shown to measure cortisol more consistently without the daily diurnal rhythm observed in blood. Commercial ELISA (enzyme-linked immunoassay) kits offer benefits over previously validated immunoassay methods but lack validation. The objective of this study was to evaluate a commercial ELISA kit (Arbor AssaysTM DetectX® Cortisol ELISA kit, K003-H1, Ann Arbor, MI, USA) and provide analytical and biologic validation of equine fecal and plasma samples. Horses (4 male, 4 female, mean ± SD: 4 ± 5 yr) were transported for 15 min with limited physical and visual contact via a livestock trailer. Blood and fecal samples were collected pre- and post-transportation. Parallelism, accuracy, and precision tests were used to analytically validate this kit. Data were analyzed using PROC MIXED in SAS 9.4. Plasma cortisol concentrations increased in response to trailering (254.5 ± 26.4 nmol/L, 0 min post-transportation) compared to pre-transportation (142.8 ± 26.4 nmol/L). FCM concentrations increased 24 h post-trailering (10.8 ± 1.7 ng/g) when compared to pre-transportation (7.4 ± 1.7 ng/g). These data support that changes in FCMs can be observed 24 h post-stressor. In conclusion, the Arbor AssaysTM DetectX® Cortisol ELISA kit is a reliable, economic option for the measurement of biologically relevant changes in cortisol in equine plasma and FCMs.

Power ultrasound-assisted enhancement of granulated blast furnace slag reactivity in cement paste

This paper introduces a first-time investigation into the impact of power ultrasound (PUS)-assisted preparation on the physicochemical and mechanical properties of cement-granulated blast furnace slag (GBFS) composite pastes. Pastes containing deposited GFBS with varying particle size fractions, partially replacing Portland cement, were prepared using PUS (ultrasonic horn tip, 20 kHz, 700 W) in pulse mode in a vertical jacketed glass sonoreactor with closed-circuit cooling. Cement paste incorporating 20 wt.% GBFS as mass substitution with varying particle size fractions was characterized by several physicochemical techniques at different curing ages. Exploring the cement and GBFS interaction induced by PUS, the compressive and flexural strength, the elastic modulus and indentation hardness, the heat of hardening, the mineral composition of hydration products, and the specific surface area BET were evaluated for a curing time of up to 28 days. The grain size distribution of GFBS and the reaction mixture's pH were measured. Both mechanical properties, heat of hydration and nanoporosity exhibited strong sensitivity to PUS treatment. Sonofragmentation of GBFS particles (especially the 125–250 μm fraction) increased with increasing sonication time, resulting in a relative increase of fraction <63 μm and a decrease of fraction >125 μm by 275 % and 60 %, respectively. Using the obtained SEM-EDS data, a simplified mechanism is proposed to explain the effects induced by PUS treatment.

Levels of matrix metalloproteinase-1 (MMP-1), MMP-2, MMP-3, osteopontin, pentraxin-3, and thymic stromal lymphopoietin in crevicular fluid samples from peri-implantitis, periodontitis, and healthy sites.

Periodontitis and peri-implantitis are chronic inflammatory diseases characterized by the destruction of supporting tissues. Despite some similarities, it is essential to understand the differences in how these diseases elicit unique host responses within the oral tissues, including the production of selected matrix metalloproteinases (MMPs) and inflammatory mediators involved in tissue remodelling. The aim of this study was to evaluate the levels of proteolytic enzymes MMP-1, MMP-2, MMP-3, as well as the inflammatory mediators osteopontin (OPN), pentraxin-3 (PTX3), and thymic stromal lymphopoietin (TSLP) in crevicular fluid samples collected from healthy, periodontitis-affected, and peri-implantitis sites. Gingival crevicular fluid (GCF) and peri-implant crevicular fluid (PICF) samples were collected from healthy and diseased teeth and implant sites of 163 patients. The MMP-1, MMP-2, MMP-3, OPN, PTX3, and TSLP levels were determined using commercially available immunoassays. A linear mixed model procedure was adopted for multilevel analyses, using biomarker levels as the outcome variable to compare two types of sites. The diagnostic accuracy of the biomarkers was evaluated by Youden's index to estimate the sensitivity, specificity and the area under curve (AUC). The levels of MMP-1, MMP-2, MMP-3, OPN, and TSLP were higher at sites with periodontitis and peri-implantitis compared to the levels at sites with healthy teeth and healthy implants. No significant differences were observed in the levels of the measured markers between the sites diagnosed with periodontitis and those diagnosed with peri-implantitis. The highest diagnostic potential at implant sites was found for MMP-2 (AUC = 0.74) and TSLP (AUC = 0.72). The highest AUC (0.82) at tooth sites was found for OPN. The findings indicate that the proteolytic enzyme MMP-2 and the cytokine TSLP might be potential biomarkers for both periodontitis and peri-implantitis, whereas the proinflammatory cytokine OPN may serve as a biomarker for periodontitis. Further studies are required to confirm the utility of these biomarkers and explore their potential clinical applications.