Pharmacokinetic modelling of orally administered cannabidiol and implications for medication control in horses.

Cannabidiol (CBD) products gain increasing popularity amongst animal owners and veterinarians as an alternative remedy for treatment of stress, inflammation or pain in horses. Whilst the use of cannabinoids is banned in equine sports, there is limited information available concerning CBD detection times in blood or urine. The aim of this study was to determine the pharmacokinetic properties of CBD following oral administration in the horse to assist doping control laboratories with interpreting CBD analytical results. Part 1: dose escalation study: Single oral administration of three escalating doses of CBD paste (0.2 mg/kg, n = 3 horses; 1 mg/kg, n = 3; 3 mg/kg, n = 5) with >7 days wash-out periods in between. Part 2: multiple dose study: oral administration of CBD paste (3 mg/kg, n = 6) twice daily for 15 days. Multiple blood and urine samples were collected daily throughout both studies. Following study part 2, blood and urine samples were collected for 2 weeks to observe the elimination phase. Concentrations of CBD, its metabolites and further cannabinoids were evaluated using gas-chromatography/tandem-mass-spectrometry. Pharmacokinetic parameters were assessed via two approaches: population pharmacokinetic analysis using a nonlinear mixed-effects model and non-compartmental analysis. AUC0-12 h and Cmax were tested for dose proportionality. During the elimination phase, the CBD steady-state urine to serum concentration ratio (Rss) was calculated. Oral CBD medication was well-tolerated in horses. Based on population pharmacokinetics, a three-compartment model with zero-order absorption most accurately described the pharmacokinetic properties of CBD. High volumes of distribution into peripheral compartments and high concentrations of 7-carboxy-CBD were observed in serum. Non-compartmental analysis identified a Cmax of 12.17 ± 2.08 ng/mL after single administration of CBD (dose: 3 mg/kg). AUC0-12 h showed dose proportionality, increase for Cmax leveled off at higher doses. Following multiple doses, the CBD terminal half-life was 161.29 ± 43.65 h in serum. Rss was 4.45 ± 1.04. CBD is extensively metabolized and shows high volumes of tissue distribution with a resulting extended elimination phase. Further investigation of the potential calming and anti-inflammatory effects of CBD are required to determine cut-off values for medication control using the calculated Rss.

Ionophore coccidiostats - disposition kinetics in laying hens and residues transfer to eggs.

Poultry production is linked with the use of veterinary medicinal products to manage diseases. Ionophore coccidiostats have been permitted for use as feed additives within the European Union (EU) for the prevention of coccidiosis in various species of poultry with except of laying hens. The presence of chemical residues in eggs is a matter of major concern for consumers' health. Despite such prohibition of use in laying hens, they were identified as the most common non-target poultry species being frequently exposed to these class of coccidiostats. Many factors can influence the presence of residues in eggs. Carryover of these class of coccidiostat feed additives in the feed of laying hens has been identified as the main reason of their occurrence in commercial poultry eggs. The physicochemical properties of individual compounds, the physiology of the laying hen, and the biology of egg formation are believed to govern the residue transfer rate and its distribution between the egg white and yolk compartments. This paper reviews the causes of occurrence of residues of ionophore coccidiostats in eggs within the EU with special emphasis on their disposition kinetics in laying hens, and residue transfer into eggs. Additional effort was made to highlight future modeling perspectives on the potential application of pharmacokinetic modeling in predicting drug residue transfer and its concentration in eggs.

The effects of cyclosporine A or activated charcoal co-administration on the pharmacokinetics of enrofloxacin in chickens.

The study aimed to investigate the possible role of efflux transporter proteins in the pharmacokinetics of enrofloxacin (ENR) in broilers in the model of co-administration of activated charcoal (AC) or cyclosporine A (CsA). The concentrations of enrofloxacin and its metabolite ciprofloxacin were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS) and population approach was used for pharmacokinetic analysis. It was found that body weight has a significant effect on the volume of distribution in the central compartment and on the systemic clearance. Oral AC increased the systemic clearance of intravenously administered ENR suggesting some role of enterohepatic recirculation. For orally administered ENR, CsA increased the area under the curve which can be explained by the inhibition of efflux transporters. Metabolism of the antibacterial drug was not affected by cyclosporine. The data suggest a role of efflux transporter proteins in the pharmacokinetics of drugs in chickens and drug-drug interactions have to be considered when substrates and modulators of these transporters are co-administered.

Predicting the efficacy of opioid sequestration by intravenous lipid emulsion using biologically relevant in vitro models of drug distribution.

Intravenous lipid emulsions (ILE), among other uses, are utilized in the treatment of poisonings caused by lipophilic substances. The body of evidence regarding the benefits of this treatment is growing but information about opioids-ILE interaction is still very scarce. In this work, the impact of ILE on the distribution of buprenorphine, fentanyl and butorphanol used in various concentrations (100-500 ng/ml) was investigated. Two different in vitro models were used: disposition of the drugs in plasma after ultracentrifugation and distribution into the simulated biophase (cell monolayer of 3T3 fibroblasts or J774.E macrophages). We confirmed the ability of ILE to sequester the three drugs of interest which results in their decrease in the aqueous part of the plasma by 34.2-38.2%, 11.7-28.5% and 6.0-15.5% for buprenorphine, fentanyl and butorphanol, respectively. Moreover, ILE affected the drug distribution to the biophase in vitro, however, in this case the drug concentration in cells decreased by 97.3 ± 3.1%, 28.6 ± 5.4% and 13.0 ± 7.5% for buprenorphine, fentanyl and butorphanol, respectively. The two models revealed notable differences in ILE's potential for drug sequestration, especially for buprenorphine. Similar, but not as pronounced tendencies were observed for the two other drugs. These discrepancies may result from the difference in protein abundance and resulting drug-protein binding in both systems. Nevertheless, the results obtained with both in vitro models correlated well with the partition coefficient (logP) values for these drugs.

Pharmacokinetics and therapeutic efficacy of levamisole in Ascaridia galli experimentally infected ducks.

This study investigated the pharmacokinetics and therapeutic efficacy of levamisole (LVS) after intravenous (i.v.) and oral administrations to healthy and Ascaridia galli-infected ducks by developing an infection model. Twenty-four two-week old ducklings were experimentally infected with A. galli. The ducks were monitored for the development of infection and after 8 weeks they were administered with LVS at a single dose of 30 mg/kg by oral or i.v. administration. Sixteen healthy ducks were subjected to the same treatment and served as control. Serial blood samples were taken for LVS determination with HPLC-UV and pharmacokinetic analysis was carried out based on the non-compartmental approach. The LVS therapeutic efficacy was determined 1 week post drug administration by intestinal worm count at necropsy. In vivo data on development of ascariasis in ducks showed that 8 weeks post inoculation the number of eggs per gram of feces reached at least 100 in each bird. After a single dose of LVS, no parasites were recovered upon necropsy. Results of the pharmacokinetic study showed no statistical differences between infected and non-infected birds for both routes of administration. The mean oral bioavailability was slightly below 50% in both experimental groups. In conclusion, the pharmacokinetics of LVS in ducks was not affected by experimentally-induced ascariasis. A single dose of LVS was found to be efficient against experimental ascariasis in ducks induced by in field isolates of A. galli.

Multifunctionality of Nanosized Calcium Apatite Dual-Doped with Li+/Eu3+ Ions Related to Cell Culture Studies and Cytotoxicity Evaluation In Vitro.

Li+/Eu3+ dual-doped calcium apatite analogues were fabricated using a microwave stimulated hydrothermal technique. XRPD, FT-IR, micro-Raman spectroscopy, TEM and SAED measurements indicated that obtained apatites are single-phased, crystallize with a hexagonal structure, have similar morphology and nanometric size as well as show red luminescence. Lithium effectively modifies the local symmetry of optical active sites and, thus, affects the emission efficiency. Moreover, the hydrodynamic size and surface charge of the nanoparticles have been extensively studied. The protein adsorption (lysozyme, LSZ; bovine serum albumin, BSA) on the nanoparticle surface depended on the type of cationic dopant (Li+, Eu3+) and anionic group (OH-, Cl-, F-) of the apatite matrix. Interaction with LSZ resulted in a positive zeta potential, and the nanoparticles had the lowest hydrodynamic size in this protein medium. The cytotoxicity assessment was carried out on the human osteosarcoma cell line (U2OS), murine macrophages (J774.E), as well as human red blood cells (RBCs). The studied apatites were not cytotoxic to RBCs and J774.E cells; however, at higher concentrations of nanoparticles, cytotoxicity was observed against the U2OS cell line. No antimicrobial activity was detected against Gram-negative bacteria with one exception for P. aeruginosa treated with Li+-doped fluorapatite.

Tylosin Dosage Adjustment Based on Allometric Scaling in Male Turkeys.

Turkeys' body weight (BW) increases 10-fold within only 2.5 months, leading to a change in the pharmacokinetics (PK) of drugs according to allometric principles. Thus, the same dosage may lead to age-dependent variability in efficacy, in particular, to treatment failure and/or selection for resistance. The study aimed to investigate whether a non-linear dosage based on a published allometric model for tylosin clearance, may optimize the internal exposure in growing turkeys. The single dose PK study was performed on turkeys aged 6, 9.5, 13 and 17 weeks (BW from 1.75 kg to 15.75 kg). Tylosin was administered intravenously (i.v.) or orally (p.o.) according to following protocols: Dose = 31.6 × BW0.58 or Dose = 158 × BW0.58, respectively. Plasma tylosin was measured using high-performance liquid chromatography and non-compartmental PK analysis was performed. The area under the curve (AUClast) after i.v. administration was 8.90 ± 1.01; 7.51 ± 1.11; 6.54 ± 1.20 and 8.01 ± 1.75 mg × h/L in 6-; 9.5-; 13- and 17-week-old turkeys, respectively. After p.o. administration AUClast was 4.80 ± 2.92; 4.60 ± 2.45; 3.00 ± 1.49 and 3.24 ± 2.00 mg × h/L in respective age groups indicating high variability. For i.v. administration, the non-linear dosage allowed to minimize the age-dependent variability in AUC. However, due to low oral bioavailability (8-12%) and resulting interindividual variability, the proposed approach may not improve tylosin efficacy in turkeys under farm conditions.

Antitrichomonal activity of metronidazole-loaded lactoferrin nanoparticles in pigeon trichomoniasis.

In recent years, increasing attention has been paid to the novel drug delivery systems to reduce the dose of the drug and avoid side effects. Metronidazole has been used for many years in the treatment of anaerobic bacterial and protozoal infections. Nanolactoferrin, a newly developed antibacterial agent originated from lactoferrin, is applied both as an active therapeutic and a drug nanocarrier. The present study describes the development and characterization of metronidazole-loaded lactoferrin nanoparticles (nano-MTZ) as well as reports their antitrichomonal activity on Trichomonas gallinae, the protozoal causative agent of pigeon trichomoniasis. The activity of the nano-MTZ is compared with the regular metronidazole formulation (MTZ) under in vitro and in vivo conditions. Additionally, cytotoxicity of the nano-MTZ to fibroblast cell line and possible hepatotoxicity in treated pigeons were evaluated. Nano-MTZ was prepared based on the thermal treatment method and the average size and surface charge of the dispersion were 30.6 nm and - 44.6 mv, respectively. No significant cytotoxicity was noted for the nano-MTZ in comparison to the MTZ. Loading efficiency in nano-MTZ was calculated as 55%. In vitro susceptibility results demonstrated 24 h 90% lethal concentration values of 4.23 and 6.64 µg/mL for MTZ and nano-MTZ, respectively. Oral treatment of the pigeons experimentally infected with T. gallinae resulted in the earlier eradication of the infection in the nano-MTZ-treated pigeons. No adverse effects on the liver function have been observed for the nano-MTZ. These findings suggest that nanolactoferrin is a promising platform for the development of novel MTZ formulations with improved antitrichomonal activity.

Lipemia in the Plasma Sample Affects Fentanyl Measurements by Means of HPLC-MS2 after Liquid-Liquid Extraction.

Examination of fentanyl levels is frequently performed in certain scientific evaluations and forensic toxicology. It often involves the collection of very variable blood samples, including lipemic plasma or serum. To date, many works have reported the methods for fentanyl detection, but none of them have provided information about the impact on the assay performance caused by an excessive amount of lipids. This aspect may be, however, very important for highly lipophilic drugs like fentanyl. To address this issue, we developed the liquid chromatography method with mass spectrometry detection and utilized it to investigate the impact of lipids presence in rabbit plasma on the analytical method performance and validation. The validation procedure, conducted for normal plasma and lipemic plasma separately, resulted in good selectivity, sensitivity and linearity. The limits of detection and quantification were comparable between the two matrices, being slightly lower in normal plasma (0.005 and 0.015 µg/L) than in lipemic plasma (0.008 and 0.020 µg/L). Liquid-liquid extraction provided a low matrix effect regardless of the lipid levels in the samples (<10%), but pronounced differences were found in the recovery and accuracy. In the normal plasma, this parameter was stable and high (around 100%), but in the lipemic matrix, much more variable and less efficient results were obtained. Nevertheless, this difference had no impact on repeatability and reproducibility. In the present work, we provided reliable, convenient and sensitive method for fentanyl detection in the normal and lipemic rabbit plasma. However, construction of two separate validation curves was necessary to provide adequate results since the liquid-liquid extraction was utilized. Therefore, special attention should be paid during fentanyl quantification that involves lipemic plasma samples purified by this technique.

Cytotoxicity Study of UV-Laser-Irradiated PLLA Surfaces Subjected to Bio-Ceramisation: A New Way towards Implant Surface Modification.

In this research we subjected samples of poly(L-lactide) (PLLA) extruded film to ultraviolet (193 nm ArF excimer laser) radiation below the ablation threshold. The modified film was immersed in Simulated Body Fluid (SBF) at 37 °C for 1 day or 7 days to obtain a layer of apatite ceramic (CaP) coating on the modified PLLA surface. The samples were characterized by means of optical profilometry, which indicated an increase in average roughness (Ra) from 25 nm for the unmodified PLLA to over 580 nm for irradiated PLLA incubated in SBF for 1 day. At the same time, the water contact angle decreased from 78° for neat PLLA to 35° for irradiated PLLA incubated in SBF, which suggests its higher hydrophilicity. The obtained materials were investigated by means of cell response fibroblasts (3T3) and macrophage-like cells (RAW 264.7). Properties of the obtained composites were compared to the unmodified PLLA film as well as to the UV-laser irradiated PLLA. The activation of the PLLA surface by laser irradiation led to a distinct increase in cytotoxicity, while the treatment with SBF and the deposition of apatite ceramic had only a limited preventive effect on this harmful impact and depended on the cell type. Fibroblasts were found to have good tolerance for the irradiated and ceramic-covered PLLA, but macrophages seem to interact with the substrate leading to the release of cytotoxic products.

High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Buprenorphine Evaluation in Plasma-Application to Pharmacokinetic Studies in Rabbits.

The precise and reliable determination of buprenorphine concentration is fundamental in certain medical or research applications, particularly in pharmacokinetic studies of this opioid. The main challenge is, however, the development of an analytical method that is sensitive enough, as the detected in vivo concentrations often fall in very low ranges. Thus, in this study we aimed at developing a sensitive, repeatable, cost-efficient, and easy HPLC analytical protocol for buprenorphine in rabbit plasma. In order to obtain this, the HPLC-MS2 system was used to elaborate and validate the method for samples purified with liquid-liquid extraction. Fragment ions 468.6→396.2 and 468.6→414.2 were monitored, and the method resulted in a high repeatability and reproducibility and a limit of quantification of 0.25 µg/L with a recovery of 98.7-109.0%. The method was linear in a range of 0.25-2000 µg/L. The suitability of the analytical procedure was tested in rabbits in a pilot pharmacokinetic study, and it was revealed that the method was suitable for comprehensively describing the pharmacokinetic profile after buprenorphine intravenous administration at a dose of 300 µg/kg. Thus, the method suitability for pharmacokinetic application was confirmed by both the good validation results of the method and successful in vivo tests in rabbits.

Allometric Optimization of Enrofloxacin Dosage in Growing Male Turkeys: Empirical Evidence for Improved Internal Exposure.

Rapid weight gain in turkeys causes a major change in the pharmacokinetics of drugs, leading to age-dependent variability in the internal exposure and, possibly, treatment failure and/or selection for antimicrobial resistance in young individuals. The aim of the study was to investigate whether a non-linear dosing protocol that accounts for the previously established allometric relation between enrofloxacin clearance and body weight (BW) may optimize the internal exposure to enrofloxacin in growing male turkeys. Enrofloxacin was administered four times, between the age of 5 and 16.5 weeks, when the turkeys' BW increased from 1.47 to 14.92 kg. Enrofloxacin was given intravenously (i.v.) or orally at the dose calculated as follows: Dose = 30 × BW0.59. After i.v. administration, the internal exposure to the drug-quantified as the area under the concentration-time curve (AUC)-was showing little age-related variation. The coefficient of variation (CV) for AUC in all individuals (15.7%) was only slightly higher than within the age groups (5.4-13.7%). After oral drug administration, CV for AUC in all individuals (22.1%) was similar as within the age groups (8.7-32.2%). These results show that intra-species allometric scaling may be efficiently implemented in the non-linear approach to enrofloxacin dosage in turkeys in order to obtain a precise internal exposure for the optimal antimicrobial effect.

Pharmacokinetics of xanthohumol in rats of both sexes after oral and intravenous administration of pure xanthohumol and prenylflavonoid extract.

BACKGROUND: Female inflorescences of hops (Humulus lupulus L.) are wildly used in the brewing industry. Hops have been also used for ages in folk medicine. Xanthohumol (XN) is a most abundant prenylated flavonoid present in hops. OBJECTIVES: To determine pharmacokinetic parameters and bioavailability of pure XN and XN given in prenylflavonoid extract obtained from spent hops (HOP). MATERIAL AND METHODS: Fifty-six Wistar rats (28 females and 28 males) were administered with XN or HOP. Xanthohumol was administered either intravenously (iv.) (10 mg/kg) or orally (per os (p.o.)) (40, 100 and 200 mg/kg). Extract obtained from spent hops was administered p.o. and its doses were based on XN content (doses were equivalent to XN dose of 40, 100 and 200 mg/kg, respectively). After administration of XN or HOP serum, XN concentration was measured at different time points (0, 0.25, 0.5, 1, 2, 4, 6, 12, 24, 48, 72, and 96 h). Non-compartmental analysis was used to assess the pharmacokinetics (PK) of XN in rats. RESULTS: The XN PK in rats after intravenous administration is characterized by extensive distribution followed by delayed elimination from the body. Enterohepatic recirculation is likely to play a role in XN PK. Some fraction of the orally administered XN reaches central compartment rapidly; however, the overall absorption is very limited and probably saturable. The formulation-dependent factors also play an important role in the bioavailability of the drug. Although the CMAX concentration was higher in female rats receiving XN orally comparing to males, the other pharmacokinetic parameters were unaffected by the rats' sex. CONCLUSIONS: The same doses of XN may be administered to male and female subjects, as its pharmacokinetics is not affected by sex.

Allometric analysis of tylosin tartrate pharmacokinetics in growing male turkeys.

BACKGROUND: Despite common use of tylosin in turkeys, the pharmacokinetic (PK) data for this drug in turkeys is limited. Within a few months of growth, PK of drugs in turkeys undergoes changes that may decrease their efficacy due to variable internal exposure. OBJECTIVES: The objective of this study was to investigate the influence of age on the PK of a single intravenous (i.v.) and oral administration of tylosin to turkeys at a dose of 10 and 50 mg/kg, respectively. METHODS: Plasma drug concentrations were measured using high-performance liquid chromatography with UV detection. The PK parameters were assessed by means of non-compartmental approach and were subjected to allometric analysis. RESULTS: During a 2.5-month-long period of growth from 1.4 to 14.7 kg, the median value for area under the concentration-time curve after i.v. administration increased from 2.61 to 7.15 mg × h/L and the body clearance decreased from a median of 3.81 to 1.42 L/h/kg. Over the same time, the median elimination half-life increased from 1.03 to 2.96 h. For the oral administration a similar trend was noted but the differences were less pronounced. Bioavailability was variable (5.76%-21.59%) and age-independent. For both routes, the plasma concentration of the major tylosin metabolite, tylosin D, was minimal. Protein binding was age-independent and did not exceed 50%. Allometric analysis indicated a relatively poor predictivity of clearance, volume of distribution and elimination half-life for tylosin in turkeys. CONCLUSIONS: Age has a significant impact on tylosin PK in turkeys and dosage adjustment may be needed, particularly in young individuals.

LiAl5O8:Fe3+ and LiAl5O8:Fe3+, Nd3+ as a New Luminescent Nanothermometer Operating in 1st Biological Optical Window.

New types of contactless luminescence nanothermometers, namely, LiAl5O8:Fe3+ and LiAl5O8:Fe3+, Nd3+ are presented for the first time, revealing the potential for applications in biological systems. The temperature-sensing capability of the nanocrystals was analyzed in wide range of temperature (-150 to 300 °C). The emission intensity of the Fe3+ ions is affected by the change in temperature, which induces quenching of the 4T1 (4G) → 6A1 (6S) Fe3+ transition situated in the 1st biological window. The highest relative sensitivity in the temperature range (0 to 50 °C) was found to be 0.82% °C (at 26 °C) for LiAl5O8: 0.05% Fe3+ nanoparticles that are characterized by long luminescent lifetime of 5.64 ms. In the range of low and high temperatures the Smax was calculated for LiAl5O8:0.5% Fe3+ to be 0.92% °C at -100 °C and for LiAl5O8:0.01% Fe3+ to be 0.79% °C at 150 °C. The cytotoxicity assessment carried out on the LiAl5O8:Fe3+ nanocrystals, demonstrated that they are biocompatible and may be utilized for in vivo temperature sensing. The ratiometric luminescent nanothermometer, LiAl5O8:Fe3+, Nd3+, which was used as a reference, possesses an Smax = 0.56%/°C at -80 °C, upon separate excitation of Fe3+ and Nd3+ ions using 266 nm and 808 nm light, respectively.

The influence of age and body weight gain on enrofloxacin pharmacokinetics in turkeys-Allometric approach to dose optimization.

Enrofloxacin is a concentration-dependent antimicrobial used in bacterial infections in poultry. During a few months of a turkey's life, pharmacokinetics of drugs undergoes substantial changes which may compromise their efficacy due to variability in internal exposure (measured by area under the concentration-time curve, AUC). The aim of this study was to describe the effects of age on the pharmacokinetics of a single intravenous (i.v.) and oral administration of enrofloxacin at a dose of 10 mg/kg to turkeys. It was found that during a 2.5-month-long period of growth from 1.4 to 14.6 kg, the AUC after i.v. administration increased almost threefold due to a significant decrease in the body clearance (from a mean of 0.76-0.28 L hr-1  kg-1 ). Over the same period, the mean elimination half-life was prolonged from 2.65 to 7.03 hr. Oral administration resulted in a similar trend in pharmacokinetic parameters. For both routes, formation of the major metabolite, ciprofloxacin, was marginal. Protein binding was not age-dependent and never exceeded 50%. Body clearance, volume of distribution and elimination half-life were subjected to an allometric analysis and a novel, nonlinear dosage protocol has been proposed to improve the internal exposure to the drug in different age groups of turkeys.

Preparation and preliminary evaluation of bio-nanocomposites based on hydroxyapatites with antibacterial properties against anaerobic bacteria.

Hexagonal nanocrystalline powders of the non-doped Ca10(PO4)6(OH)2 as well as activated with Ag+ and Eu3+ ions were synthesized by using different wet chemistry methods. Moreover, the obtained hydroxyapatite was loaded with Ag0, as well as nitroimidazole antimicrobials: metronidazole and tinidazole. The structural properties of the products were analyzed by X-ray diffraction (XRD), scanning (SEM) and transmission (TEM) electron microscopy as well as infrared (IR) and Raman spectroscopy. The photoluminescence properties of the Eu3+ and Ag+ co-doped Ca10(PO4)6(OH)2 were characterized via the PL emission, excitation spectra and the luminescence decay curve. The antimicrobial activity of the obtained materials against Prevotella bivia and Parabacteroides distasonis was studied. The cytotoxicity assessment was carried out on the human osteosarcoma cell line (U2OS) as well as human red blood cells (RBC). The choice of the in vitro model was based on the fact that U2OS is a cancer cell line derived from bone tissue which is rich in apatites that play a pivotal role in the extracellular matrix formation. RBCs are the most abundant blood cells and they are used as a cell model in the study of biocompatibility of new prepared biocompounds with potential medical applications. The obtained multifunctional materials do not exhibit the haemolytic activity, therefore, they could be used as a promising antimicrobial agent and for anaerobic bacteria.

Exercise-induced changes in skin temperature and blood parameters in horses.

The aim of the study was to assess the effects of training on haematological and biochemical blood parameters as well as on the changes in body surface temperature in horses. In order to identify the predictive value of surface temperature measurements as a marker of animal's performance, their correlations with blood parameters were investigated. The study was carried out on nine horses divided into two groups: routinely ridden and never ridden. Infrared thermography was used to assess surface temperature changes before (BT) and just after training (JAT) on a treadmill. Seven regions of interest (ROIs) located on the neck, shoulder, elbow, back, chest, gluteus and quarter were analysed. The blood samples were taken BT, JAT and 30 min after training (30AT). Haematological parameters including white blood cells, lymphocytes (LYMs), monocytes (MONOs), granulocytes (GRAs), eosinophils (EOSs), haematocrit (HCT) and platelets (PLTs) as well as biochemical parameters such as glucose (GLUC), urea, Na + , K + and Ca 2 + , and creatine phosphokinase (CPK) were analysed. Our results indicated a significant increase in surface temperature JAT ( p = 0.043 ) in the neck, shoulder, elbow, gluteus and quarter in routinely ridden horses. Significant changes in EOS ( p = 0.046 ) and HCT ( p = 0.043 ) in the case of the never-ridden and routinely ridden group, respectively, were found between the times of blood collection. In addition, there was a significant effect of the horse group and the time of blood collection on the CPK activity ( p = 0.025 to p = 0.045 ) and urea concentrations ( p = 0.027 to p = 0.045 ). In the routinely ridden horses, there were significant correlations between the changes in MONO ( ρ = 0.40 ), GRA ( ρ = - 0.40 ), PLT ( ρ = - 0.77 ), HCT ( ρ = - 0.36 ), GLUC ( ρ = 0.56 ) and urea ( ρ = 0.56 ) and the total ROI temperature changes. Moreover, significant correlations between the changes in MONO ( ρ = - 0.86 ) , EOS ( ρ = - 0.65 ), GLUC ( ρ = 0.85 ), urea ( ρ = 0.85 ), Na + ( ρ = 0.59 ) and K + ( ρ = - 0.85 ) and the total ROI temperature changes were found in never-ridden horses. Different changes in body surface temperature and blood parameters in routinely ridden and never-ridden horses could be associated with different conditioning and performance. A significantly higher surface temperature in routinely ridden horses, as well as the dynamics of changes in HCT, CPK and urea after training indicate better performance of these horses. Significant correlations between MONO, GLUC, and urea and a total ROI surface temperature as well as a negative correlation between MONO and the total ROI temperature in never-ridden horses indicated poor performance.

Protective effects of betulinic acid on intestinal mucosal injury induced by cyclophosphamide in mice.

BACKGROUND: Betulinic acid (BA) is a plant-derived pentacyclic triterpenoid with a variety of biological activities. The purpose of this study was to assess the potential protective role of BA against intestinal mucosal injury induced by cyclophosphamide (CYP) treatment. METHODS: Mice were pretreated with BA daily (0.05, 0.5, and 5.0 mg/kg) for 14 days, then injected intraperitoneally with CYP (50 mg/kg) for 2 days. RESULTS: BA pretreatment reduced the contents of malondialdehyde (MDA) and glutathione (GSH), decreased the activity of superoxide dismutase (SOD) in small intestine, increased villus hight/crypt depth ratio and restored the morphology of intestinal villi in CYP-induced mice. Moreover, BA pretreatment could significantly down-regulate the levels of pro-inflammatory cytokines interleukin-5 (IL-5), IL-17, IL-12 (P70) and tumor necrosis factor α (TNF-α), reduced production of chemokines macrophage inflammatory protein-1α (MIP-1α), macrophage inflammatory protein-1ß (MIP-1ß) and regulated upon activation, normal T-cell expressed and secreted (RANTES), and enhanced the levels of anti-inflammatory such as IL-2 and IL-10 in serum, and decreased the mRNA expressions of IL-1ß and TNF-α in intestine of CYP-induced mice. Furthermore, RT-PCR demonstrated that BA improved intestinal physical and immunological barrier in CYP-stimulated mice by enhancing the mRNA expressions of zonula occluden 1 (ZO-1) and Claudin-1. CONCLUSIONS: BA might be considered as an effective agent in the amelioration of the intestinal mucosal resulting from CYP treatment.