M1 Concentrations Research Articles

Biological System Responses of Dairy Cows to Aflatoxin B1 Exposure Revealed with Metabolomic Changes in Multiple Biofluids.

Research on mycotoxins now requires a systematic study of post-exposure organisms. In this study, the effects of aflatoxin B1 (AFB1) on biofluids biomarkers were examined with metabolomics and biochemical tests. The results showed that milk concentration of aflatoxin M1 changed with the addition or removal of AFB1. AFB1 significantly affected serum concentrations of superoxide dismutase (SOD) and malon dialdehyde (MDA), SOD/MDA, and the total antioxidant capacity. Significant differences of volatile fatty acids and NH3-N were detected in the rumen fluid. Eighteen rumen fluid metabolites, 11 plasma metabolites, and 9 milk metabolites were significantly affected by the AFB1. These metabolites are mainly involved in the pathway of amino acids metabolism. Our results suggest that not only is the study of macro-indicators (milk composition and production) important, but that more attention should be paid to micro-indicators (biomarkers) when assessing the risks posed by mycotoxins to dairy cows.

Single- and multiple-dose pharmacokinetics of the peripheral non-narcotic antitussive moguisteine in healthy Chinese volunteers

BackgroundMoguisteine is a non-narcotic peripheral antitussive drug that has been effective and well-tolerated in clinical studies. The aim of the present work was to investigate the pharmacokinetics of moguisteine given as single or multiple doses to healthy Chinese subjects. MethodsIn Stages 1–3 of this study, 12 healthy Chinese subjects (6 males and 6 females) participated in a randomized, open-label, single-dose, 3-period, 3-way crossover study, with a 24-h washout period between each treatment. Eligible subjects were randomized to receive a single dose of 100, 200 or 400 mg moguisteine. Blood was sampled before and up to 10 h after administration. In those receiving 200 mg moguisteine, urine was sampled at intervals of 0–2, 2–4, 4–6, 6–10, and 10–24 h. In Stage 4, subjects received a moguisteine tablet containing 200 mg three times daily for five consecutive days. Blood was sampled for up to 10 h after the last dose. HPLC-tandem mass spectrometry was used to determine concentrations of the moguisteine metabolite M1 in serum, while HPLC-UV was used to determine concentrations of M1 in urine. Safety of the dosing schedules was assessed based on physical examination, recording of adverse events, 12-lead electrocardiography, and laboratory tests. ResultsAll subjects completed all four stages of the study. M1 was detectable at the shortest time points after moguisteine administration; the time to achieve peak concentration was 0.5–1.0 h in single dosing and 1.5 h in multiple dosing. Elimination half-life (t1/2) was 0.91–1.54 h in single dosing and 1.57 h in multiple dosing. AUC increased roughly proportionally with dose, while Cmax increased much more gradually with dose. During 5-day dosing of three tablets per day, a steady state concentration was reached on day 3, and the mean accumulation ratio was 0.87. At 24 h after a single dose of 200 mg moguisteine, approximately 34.0% of the resulting M1 was recovered in urine. Pharmacokinetics of moguisteine did not differ significantly between men and women, except among those receiving a single dose of 100 mg (P < 0.05). Mild adverse events (nausea, loose stool, abdominal distention, or dizziness) occurred in six subjects and resolved without treatment, while no serious adverse events were observed. ConclusionMoguisteine was safe and well-tolerated by our healthy subjects, and it exhibited dose linearity but not proportionality when a single dose of 100–400 mg was given. M1 did not accumulate in subjects after multiple doses of moguisteine.

Pharmacokinetics and Safety of Enzalutamide in Healthy Chinese Male Volunteers

PurposeThis open-label, single-dose study evaluated the pharmacokinetic profiles of enzalutamide and its major metabolites and the safety of enzalutamide in healthy, Chinese male volunteers. MethodsFourteen volunteers (median age, 28.5 years) received a single oral dose of enzalutamide (160 mg) under fasting conditions on day 1 and were followed for 50 days. Pharmacokinetic profiles were obtained for enzalutamide and its major metabolites, carboxylic acid metabolite (M1; inactive metabolite) and N-desmethyl enzalutamide (M2; active metabolite), on day 1 up to 1176 hours (49 days). Safety data were also collected. FindingsEnzalutamide plasma concentration rapidly increased (median Tmax, 1.5 hours) followed by a slow decrease (mean t½, 90.7 hours). M1 and M2 plasma concentrations increased gradually with a median Tmax of 72.0 and 121 hours, respectively. M1 and M2 mean metabolite-to-parent ratios were 0.2 and 1.3, respectively. Mean AUC0–∞ of enzalutamide plus M2 was 828 μg h/mL versus 368 μg h/mL for enzalutamide alone. Mean t½, maximum concentration, and Tmax of enzalutamide plus M2 were comparable with those of enzalutamide. Drug-related treatment-emergent adverse events were reported in 4 men (28.6%): 1 each of upper respiratory tract infection, chest discomfort, increased blood bilirubin, and decreased white blood cell count. No deaths or serious treatment-emergent adverse events were observed. ImplicationsThe pharmacokinetic profiles of enzalutamide, M1, M2, and enzalutamide plus M2 in healthy Chinese men were generally consistent with those in white men. No new safety concerns were found. Chinese Clinical Trial Registration identifier: CTR20150635.

Towards Understanding the Anticorrosive Mechanism of Novel Surfactant Based on Mentha pulegium Oil as Eco-friendly Bio-source of Mild Steel in Acid Medium: a Combined DFT and Molecular Dynamics Investigation

Today, due to the increasingly stringent European directives concerning the use of molecules with certain toxicities towards the environment or their users, the essential oils, extracts, and molecules derived from plants exhibiting the characteristic of being biodegradable can be considered as a source of green corrosion inhibitors instead of harmful synthetic chemicals. The present work was devoted to testing the essential oil extracted from Mentha pulegium leaves(M1) as a corrosion inhibitor for C-steel in 1 mol/L HCl solution using both electrochemical techniques and gravimetric measurements for the evaluation of the inhibition efficiencies at different temperatures. The results obtained showed that the inhibition efficiency increased with an increase in M1 concentration to reach a maximum value of 92.21%. We sought to determine the molecule responsible for this high efficiency, starting with the analysis of oil chemical composition by gas chromatography coupled with mass spectrometry. This analysis revealed that menthol(M2) and isomenthol(M3) were the principal constituents. In order to identify the molecule responsible for the inhibition and explain the protection mechanism involved, quantum chemical calculations and Monte Carlo simulations were used to explain the interaction of menthol, the major constituent of M1 with the Fe-surface. To practically confirm these results, we studied the action of 1 mol/L HCl on steel with and without the addition of M2 by both methods(gravimetric and electrochemical study). A very high efficiency was obtained, an efficiency of 94.90% at 10‒3 mol/L, which was retained for a long exposure time, and slightly decreased in function of temperature. Finally, a good correlation between the experimental data, theoretical calculations, and SEM studies was obtained, which denied that the M1 efficiency was only a result of a synergy effect and confirmed the high efficiency of Mentha oil and its main component(menthol) as a strong ecological inhibitor of corrosion.

Endogenous Omega (n)-3 Fatty Acids in Fat-1 Mice Attenuated Depression-Like Behavior, Imbalance between Microglial M1 and M2 Phenotypes, and Dysfunction of Neurotrophins Induced by Lipopolysaccharide Administration.

n-3 polyunsaturated fatty acids (PUFAs) have been reported to improve depression. However, PUFA purities, caloric content, and ratios in different diets may affect the results. By using Fat-1 mice which convert n-6 to n-3 PUFAs in the brain, this study further evaluated anti-depressant mechanisms of n-3 PUFAs in a lipopolysaccharide (LPS)-induced model. Adult male Fat-1 and wild-type (WT) mice were fed soybean oil diet for 8 weeks. Depression-like behaviors were measured 24 h after saline or LPS central administration. In WT littermates, LPS reduced sucrose intake, but increased immobility in forced-swimming and tail suspension tests. Microglial M1 phenotype CD11b expression and concentrations of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-17 were elevated, while M2 phenotype-related IL-4, IL-10, and transforming growth factor (TGF)-β1 were decreased. LPS also reduced the expression of brain-derived neurotrophic factor (BDNF) and tyrosine receptor kinase B (Trk B), while increasing glial fibrillary acidic protein expression and pro-BDNF, p75, NO, and iNOS levels. In Fat-1 mice, LPS-induced behavioral changes were attenuated, which were associated with decreased pro-inflammatory cytokines and reversed changes in p75, NO, iNOS, and BDNF. Gas chromatography assay confirmed increased n-3 PUFA levels and n-3/n-6 ratios in the brains of Fat-1 mice. In conclusion, endogenous n-3 PUFAs may improve LPS-induced depression-like behavior through balancing M1 and M2-phenotypes and normalizing BDNF function.

Designing Metal-Free Frustrated Lewis Pairs Catalyst for the Efficient Dehydrogenation of Ammonia Borane.

Ammonia borane (AB) has been in the spotlight for the chemical storage of hydrogen over the past decade. However, the development of methods for efficient and controlled hydrogen release from AB under mild conditions is still underway. Herein, using density functional theory (DFT) computations, we designed a metal-free frustrated Lewis pair (FLP) catalyst o-(BPh2 )C6 H4 (NiPr2 ) (M1) that can efficiently dehydrogenate AB to release more than two equivalents of H2 under mild conditions. Catalyst M1 can dehydrogenate not only AB to H2 N=BH2 (AOB) and H2 , but also oligomers of AOB with rather low free-energy barriers. The high dehydrogenation activity of M1 is the key of new oligomerization routes to the efficient dehydrogenation of AB to borazine (BZ) or H2 B-(NH=BH)n -NH2 (PIB) and finally to polyborazylene (PBZ) so that more than two equivalents of H2 can be released. A first-principle kinetic Monte Carlo (KMC) study reveals that the activity of our catalytic system can be tuned by varying the initial concentration of M1 and AB. This work can guide the design of catalyst for the highly efficient utilization of AB as a hydrogen storage material.

The prevalence of aflatoxin M1 in milk of Middle East region: A systematic review, meta-analysis and probabilistic health risk assessment

The current investigation was undertaken to take a review of the performed studies regarding the concentration and prevalence of aflatoxin M1 (AFM1) of the consumed cow milk in the Middle East. In this context, all available studies published in databases include Science Direct, PubMed, Scopus, and Web of Science among 1995 to December 2017; were screened accordingly. Also, the carcinogenic risk was estimated by calculating hazard index (HI) using the Monte Carlo simulation (MCS). The result of conducted meta-analysis for 49 articles containing 7484 data indicated that the rank order of type of milk based on the concentration of aflatoxin M1 was Ultra-high temperature processing (UHT) milk (82.57 ng/kg) > raw milk (60.37 ng/kg) > pasteurized milk (PAS) (45.81 ng/kg). The pooled concentration of aflatoxin M1 in raw and UHT milk was higher than EC (European Committee, 50 ng/kg) standard limit. The rank order of countries based on the concentration of aflatoxin M1 in raw milk was Syria > Turkey > Iran > Egypt > Lebanon > Palestine; pasteurized milk, Turkey > Iran > Lebanon; and UHT milk, Iran > Turkey > Saudi Arabia. The overall prevalence of aflatoxin M1 in the raw milk of Iran, Turkey, Lebanon, Palestine, Egypt, and Syria was identified as 76%, 12%, 67%, 85%, 38%, and 14%; pasteurized milk, in the Iran, Lebanon, and Turkey was 77%, 36%, and 11%; and finally UHT milk in Iran, Saudi Arabia, and Turkey was 81%, 82%, and 62%, respectively. HI in the adult's consumer's raw milk in the Iran, Turkey, Syria, Palestine, Lebanon, and Egypt were calculated as 0.26, 0.47, 0.52, 0.34, 0.23 and 0.18; However, the HI for adult consumers of pasteurized milk in the Iran, Turkey, and Lebanon were 0.28, 0.31 and 0.11. Also, the measured HI for adult consumers of UHT milk in the Saudi Arabia, Turkey, and Iran was 0.20, 0.33 and 0.50, respectively. The obtained HI for consumers of raw milk in the children age group of the Iran, Turkey, Syria, Palestine, Lebanon, and Egypt were 1.03,2.20, 2.42, 1.59, 1.05, and 0.84. The calculated HI for consumers of pasteurized milk in the children age group of the pasteurized milk in of Iran, Turkey, and Lebanon were 1.30, 1.56, and 0.50. Finally, in term of UHT milk in Saudi Arabia, Turkey, and Iran were 0.94, 1.44 and 2.35, respectively. Unlike Adults, children consumers in the several Middle East countries are at considerable cancer risk due to consumption of raw, pasteurized and UHT milk contain AFM1(HI > 1).

Rapid LC-MS/MS method for quantification of vinorelbine and 4-O-deacetylvinorelbine in human whole blood suitable to monitoring oral metronomic anticancer therapy.

A rapid and sensitive LC-MS/MS method for therapeutic drug monitoring oral vinorelbine (VRL) metronomic anticancer chemotherapy has been developed and validated. Analysis of VRL and its main active metabolite 4-O-deacetylvinorelbine (M1) was performed in whole blood matrix. Both analytes were extracted by protein precipitation and separated on an Onyx monolith C18 , 50 × 2 mm column then quantified by positive electrospray ionization and multiple reaction monitoring mode. The LLOQ was 0.05 ng/mL for both VRL and M1. Linearity was up to 25ng/mL with R2 ≥ 0.994. The intra- and inter-assay precisions were ≤ 11.6 and ≤ 10.4% while the ranges of accuracy were [-8.7%; 10.3%] and [-10.0; 7.4%] for VRL and M1, respectively. The clinical suitability of the method has been proved by the determination of the CTrough blood concentrations of VRL and M1 in 64 nonsmall cell lung cancer elderly patients. The analytical performance of the assay was suitable for pharmacokinetic monitoring of VRL and M1, allowing the personalization of the VRL metronomic treatments.

Correlation between plasma concentrations of tramadol and its metabolites and the incidence of seizure in tramadol-intoxicated patients.

Seizure is one of the important symptoms of tramadol poisoning, but its causes are still unknown. The aim of this study is to find a relationship between tramadol and the concentrations of its metabolites versus the incidence of seizures following the consumption of high doses of tramadol. For this purpose, the blood samples of 120 tramadol-intoxicated patients were collected. The patients were divided in two groups (seizure and non-seizure). The concentrations of tramadol and its metabolites (M1, M2 and M5) were measured by using a high-performance liquid chromatography method. The relationship between tramadol and the levels of its metabolites and seizure incidences was also investigated. In 72% of the patients, seizures occurred in the first 3 h after the ingestion of tramadol. The seizure incidences were significantly correlated with the patients' gender, concentrations of tramadol, M1 and M2 and the history of previous seizures (p<0.001). The average concentration of M2 was significantly higher in males (p=0.003). A previous history of the use of sedative-hypnotics and the co-ingestion of benzodiazepines and other opioids were shown to significantly decrease the rate of seizure. The rate of seizure was directly related to the concentrations of tramadol and its metabolites. Higher M2 concentration in males can be considered a reason for increased incidences of seizures in males. The plasma concentration of M1 affected the onset of seizure. Therefore, it can be concluded that differences in the levels of the metabolites can affect the threshold of seizure in tramadol-intoxicated patients.

Concentration of Aflatoxin M1 and selected heavy metals in mother milk samples from Pakistan

Mother milk is the primary food source for neonates that if contaminated with certain toxic compounds may result in lifelong complications in the breastfeed infants. Present study was designed to evaluate the concentration of Aflatoxin M1 (AFM1) and the four most toxic heavy metals i.e. lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As) in mother milk samples. AFM1 was found in the range of <0.001–0.044 μg/L and 6.4% mother milk samples were found above the EU permissible limit (0.025 μg/L). Pb and Cd concentrations were found above the normal ranges proposed by WHO while Hg was found slightly above the WHO proposed level.

Pharmacokinetic and Safety Evaluation of a Transscleral Sustained Unoprostone Release Device in Monkey Eyes.

We evaluate the ocular tissue distribution and retinal toxicity of unoprostone (UNO) during 12 months, after transscleral sustained-UNO administration using a drug delivery device in monkey eyes. The device consisted of a reservoir, controlled-release cover, and a drug formulation of photopolymerized polyethylene glycol dimethacrylate. Six mg UNO was loaded into the device (length, 17 mm; width, 4.4 mm; height, 1 mm). The concentrations of M1, a primary metabolite of UNO, in the retina, choroid, vitreous, lens, aqueous humor, iris, ciliary body, and plasma were determined by liquid chromatography-tandem mass spectrometry at 3, 6, and 12 months after implantation. Retinal toxicity was evaluated by electroretinography (ERG), optical coherence tomography (OCT), and IOP at preimplantation, and at 6, 9, and 12 months after implantation. Focal ERGs were performed at 9 and 12 months after implantation. M1 was detected in the choroid and retina with maximum peaks of 243.2 and 8.41 ng/g at 6 months, respectively. M1 in the ciliary body and iris was detected with maximum peaks of 7.66 and 10.4 ng/g at 6 and 12 months, respectively. Less than 1 ng/mL or ng/g of M1 was detected in the aqueous humor, vitreous, and lens. No changes were observed in retinal function as assessed by ERG, IOP, or macula thickness and retinal histology by OCT examinations during the 12-month period. No differences in focal ERG amplitudes, especially in the macula, were observed. The device provided intraocular sustained delivery of UNO for 12 months without producing severe retinal toxicity.

Reduction in aflatoxin M1 concentration during production and storage of selected fermented milks

This study focused on monitoring changes in aflatoxin M1 (AFM1) concentrations during production and storage of different fermented milks using selected probiotic and nonprobiotic combined cultures. Milk samples intended for fermentation were intentionally contaminated by adding a standard of AFM1. All of the tested cultures caused remarkable reductions in AFM1 concentrations during the fermentation process. Probiotic cultures were more effective than nonprobiotic cultures, with Lactobacillus caseiLC‐01 strain being the most efficient, achieving a reduction level of approximately 58%. Among the nonprobiotic cultures, yoghurt culture YC‐380 was the most efficient, with a reduction level of approximately 41%.

Physiological responses to a yeast and clay-based adsorbent during an aflatoxin challenge in Holstein cows

The objective of this study was to determine the effects of 2 different adsorbents (AD) composed of yeast fractions and bentonite (AD1 and AD2) during an aflatoxin (AF) challenge on the health and performance of lactating dairy cows. Lactating Holstein cows [(n=76); BW (mean±SD)=698±72kg; DIM=153±83 d] were assigned to 1 of 5 treatments in a randomized complete block design. The trial lasted 28 d and measurements were made from d 22 to 28. From d 22 to 24 cows received an AF challenge (100μg of AFB1/kg of diet DM administered orally). Treatments were: control (CON), no AD or AF; positive control (POS), no AD plus AF challenge; 30g per cow per day of AD1 and AF challenge (P30); 60g per cow per day of AD1 and AF challenge (P60); and 60g per cow per day of AD2 and AF challenge (PROT). The appearance and disappearance of AF excreted in milk was tested at each milking from d 22 to 28 using SNAP tests (SNP; IDEXX, Inc.). Blood was sampled on d 22 and 26 (n=2 per cow), and analyzed for superoxide dismutase (SOD) concentration. Milk samples from d 22 to 26 were analyzed for AFM1 concentrations by HPLC. Fecal samples collected from the rectum on d 22 and 24 were analyzed for AFB1 concentrations via HPLC. Statistical analysis was performed using the MIXED procedure of SAS. A quadratic treatment effect (P<0.001) was observed for plasma SOD concentrations at 2.77, 1.99, and 1.97±0.05U/mL for POS, AD30, and AD60 treatments, respectively. Aflatoxin M1 transfer (11.4 and 0.00±1.60g/kg), excretion (29.52 and 0.00±4.58μg/d), and concentrations in milk (0.76 and 0.00±0.16μg/kg) were greater for POS than CON, respectively (P<0.001) but no differences were observed among other treatments. A tendency for a quadratic treatment effect (P=0.08) was observed for fecal AFB1 concentrations at 6.78, 8.55, and 5.07±1.17μg/kg for the POS, P30, and P60 treatments, respectively. Oral supplementation of yeast and bentonite clay-based adsorbents during an AF challenge resulted in quadratic changes in plasma SOD and fecal AFB1 concentrations; however, no differences were observed for DMI or milk yield. In conclusion, yeast cell wall and bentonite-based adsorbent may be beneficial in reducing inflammation during an AF challenge but there were no treatment differences regarding its adsorbent capacity.

Immobilized aptamer on gold electrode senses trace amount of aflatoxin M1

An electrochemical aptasensor for detection of trace amounts of aflatoxin M1 was developed. This required immobilization of aptamer on screen printed gold electrode comprising of working electrode, counter electrode and reference electrode and was achieved by sequentially layering dithiodipropionic acid, streptavidin and biotinylated-tetraethylene glycol-aptamer. Immobilization of aptamer was monitored by cyclic voltammetry. Peak current in square wave voltammogram was inversely related to logarithmic concentration of aflatoxin M1. Dynamic range of sensor was 1–105 ppt aflatoxin M1. Sensor can be regenerated by treating electrode with 10% sodium dodecyl sulfate or 40 mM tris-HCl (pH 8.0) containing 10 mM ethylenediaminetetraacetic acid and 0.02% tween-20.

Preliminary pharmacokinetics of tramadol hydrochloride after administration via different routes in male and female B6 mice

Objective1) To determine the pharmacokinetics of tramadol hydrochloride and its active metabolite, O-desmethyltramadol (M1), after administration through different routes in female and male C57Bl/6 mice; 2) to evaluate the stability of tramadol solutions; and 3) to identify a suitable dose regimen for prospective clinical analgesia in B6 mice. Study designProspective, randomized, blinded, parallel design. AnimalsA total of 18 male and 18 female C57Bl/6 mice (20–30 g). MethodsMice were administered 25 mg kg−1 tramadol as a bolus [intravenously (IV), intraperitoneally (IP), subcutaneously (SQ), orally per gavage (OSgavage)] over 25 hours [orally in drinking water (OSwater) or Syrspend SF (OSSyrsp)]. Venous blood was sampled at six predetermined time points over 4 to 31 hours, depending on administration route, to determine tramadol and M1 plasma concentrations (liquid chromatography and tandem mass spectrometry detection). Pharmacokinetic parameters were described using a noncompartmental model. The stability of tramadol in water (acidified and untreated) and Syrspend SF (0.20 mg mL−1) at ambient conditions for 1 week was evaluated. ResultsAfter all administration routes, Cmax was >100 ng mL−1 for tramadol and >40 ng mL−1 for M1 (reported analgesic ranges in man) followed by short half-lives (2–6 hours). The mean tramadol plasma concentration after self-administration remained >100 ng mL−1 throughout consumption time. M1 was found in the OSSyrs group only at 7 hours, whereas it was detectable in OSwater throughout administration. Tramadol had low oral bioavailability (26%). Short-lasting side effects were observed only after IV administration. Water and Syrspend SF solutions were stable for 1 week. Conclusions and clinical relevance1) At the dose administered, high plasma concentrations of tramadol and M1 were obtained, with half-life depending on the administration route. 2) Plasma levels were stable over self-consumption time. 3) Solutions were stable for 1 week at ambient conditions.

TSG-6 is highly expressed in human abdominal aortic aneurysms

BackgroundThe formation of abdominal aortic aneurysms (AAA) is characterized by a dominance of proinflammatory forces that result in smooth muscle cell apoptosis, extracellular matrix degradation, and progressive diameter expansion. Additional defects in the antiinflammatory response may also play a role but have yet to be fully characterized. TSG-6 (TNF-stimulated gene-6) is a potent antiinflammatory protein involved in extracellular matrix stabilization and cell migration active in many pathological conditions. Here, we describe its role in AAA formation. MethodsBlood and/or aortic tissue samples were collected from organ donors, subjects undergoing elective AAA screening, and open surgical AAA repair. Aortic specimens collected were preserved for IHC or immediately assayed after tissue homogenization. Protein concentrations in tissue and plasma were assayed by ELISA. All immune cell populations were assayed using FACS. In vitro, macrophage polarization from monocytes was performed with young, healthy donor PBMCs. ResultsTSG-6 was found to be abnormally elevated in both the plasma and aortic wall of patients with AAA compared with healthy and risk-factor matched non-AAA donors. We observed the highest tissue concentration of TSG-6 in the less-diseased proximal and distal shoulders compared with the central aspect of the aneurysm. IHC localized most TSG-6 to the tunica media with minor expression in the tunica adventitia of the aortic wall. Higher concentrations of both M1 and M2 macrophages where also observed, however M1/M2 ratios were unchanged from healthy controls. We observed no difference in M1/M2 ratios in the peripheral blood of risk-factor matched non-AAA and AAA patients. Interesting, TSG-6 inhibited the polarization of the antiinflammatory M2 phenotype in vitro. ConclusionsAAA formation results from an imbalance of inflammatory forces causing aortic wall infiltration of mononuclear cells leading to the vessel breakdown. In the AAA condition, we report an elevation of TSG-6 expression in both the aortic wall and the peripheral circulation.

Pharmacokinetics and Pharmacodynamics Evaluation of Tramadol In Thermoreversibles Gels

In this study, new drug delivery formulations of tramadol (TR) in thermoreversible gels were studied with regard to the pre-clinical pharmacokinetics (PK) and pharmacodynamics (PD). The poloxamer (PL)-TR systems were prepared by direct dispersion of the drug (20 mg.mL-1) in solutions with PL 407 and PL 188. The formulations used in this study were: F1- TR 2% in aqueous solution; F2- PL 407 (20%) + PL 188 (10%) + TR 2%; F3- PL 407 (25%) + PL 188 (5%) + TR 2% and F4 - PL 407 (20%) + TR 2%. New Zealand white rabbits were divided in four groups (n=6) and treated by subcutaneous route with F1, F2, F3 or F4 (10 μg.kg-1). Blood samples (1 mL) from an ear vein were collected pre dose (0 min) and at 15, 30, 45, 60, 90, 120, 180, 240, 300, 360, 420 and 480 minutes after the injection of formulations. TR and its main metabolite (M1) plasma levels were determinate using a triple stage quadrupole mass spectrometer. PD evaluation was performed with the measurement of both pupils’ diameters using callipers in the same periods of blood collection. F2 showed higher TR plasma concentration than all other formulations after 180 minutes and presented lower M1 concentrations at almost all evaluated periods (P<0.05). Areas under the curve and cleareance of TR in F2 presented differences compared to F1 (P<0.05). F2 presented significant Pearson correlation between the enhancement of TR / M1 concentrations and miosis (P<0.05). Thus, the association of PL407 (20%) and PL188 (10%) in F2 altered pharmacokinetics and pharmacodynamics of TR, since this formulation was effective to enhance the bioavailability and this effect was correlated with a more intense biologic effect.

The effects of ketoconazole and cimetidine on the pharmacokinetics of oral tramadol in greyhound dogs.

Tramadol is administered to dogs for analgesia but has variability in its extent of absorption, which may hinder its efficacy. Additionally, the active opioid metabolite (M1) occurs in low concentrations. The purpose of this study was to determine if administration of oral tramadol with suspected metabolism inhibitors (ketoconazole, cimetidine) would lead to improved bioavailability of tramadol and M1. Six healthy Greyhounds were included. They were administered tramadol orally and intravenously, M1 intravenously, oral tramadol with oral ketoconazole and oral tramadol with oral cimetidine. Oral tramadol bioavailability was low (2.6%). Ketoconazole and cimetidine significantly increased tramadol bioavailability to 18.2% and 20.3%, respectively. The mean maximum plasma concentration of tramadol alone was 22.9ng/ml, and increased to 109.9 and 143.2μg/ml with ketoconazole and cimetidine, respectively. However, measured tramadol plasma concentrations were below the minimum concentration considered effective in humans (228μg/ml). In all treatment groups, measured M1 concentrations (<7μg/ml) were below concentrations associated with efficacy in humans. To conclude, tramadol and M1 concentrations were low and variable in dogs after oral dosing of tramadol, even in combination with cimetidine or ketoconazole, but effective concentrations in dogs have not been defined.

Pharmacokinetics (PK) and safety of enzalutamide in healthy Chinese males.

e14073 Background: Enzalutamide (ENZ), an androgen receptor inhibitor, is approved in over 60 countries except China for use in metastatic castration-resistant prostate cancer. The PK of ENZ have been assessed in healthy Caucasian males (Gibbons 2015). This open-label, single-dose study evaluated the PK of ENZ and its major metabolites, and safety of ENZ, in healthy Chinese male volunteers. Methods: Non-elderly healthy Chinese male volunteers received a single oral dose of ENZ (160 mg) under fasting conditions on day 1 and were followed for 50 days. Full PK profiles were obtained for ENZ and its major metabolites, carboxylic acid metabolite (M1; inactive metabolite) and N-desmethyl enzalutamide (M2; active metabolite), on day 1 up to 1176 hours (49 days). Safety data were collected from screening until the end of study visit. Results: 14 healthy Chinese male volunteers were enrolled and received a single ENZ dose (mean age, 29.0 years; mean BMI, 21.7 kg/m2). All volunteers receiving study drug were included in the PK and safety analyses; two subjects withdrew from the study during the study period. ENZ plasma concentration rapidly increased (median tmax of 1.5 hours) followed by a slow decrease (mean t1/2 of 90.7 hours). Plasma concentrations of M1 and M2 increased gradually with a median tmax of 72.0 and 121 hours, respectively. M1 and M2 mean metabolite to parent ratio was 0.206 and 1.32, respectively. Mean AUC0-inf of the sum of ENZ and M2 was 828 μg·h/mL compared to 368 μg·h/mL for ENZ. Cmax and tmax of the sum of ENZ and M2 were comparable to ENZ. Mean t1/2 of the sum of ENZ and M2 was 155 hours, which was similar to that of M2 (162 hours). Treatment-emergent adverse events (TEAEs) were reported in seven out of 14 males (50.0%). Possible/probable drug-related TEAEs were reported in four males (28.6%); one each of upper respiratory tract infection, chest discomfort, increased bilirubin, and decreased white blood cell count. No deaths, serious TEAEs, or TEAEs of grade ≥3 were observed. Conclusions: The PK profiles of ENZ, M1, M2, and the sum of ENZ and M2 in healthy Chinese males were generally consistent with Caucasian males. No clinically relevant safety concerns were reported. Safety and PK profiles indicate that ENZ 160 mg/day is appropriate in a Chinese population.

Abstract 109: Tumor Necrosis Factor Inducible Gene 6 Protein (TSG-6) is Highly Expressed in Human Abdominal Aortic Aneurysms

Objective: The formation of an abdominal aortic aneurysm (AAA) is characterized by a dominance of pro-inflammatory forces that result in smooth muscle cell apoptosis, extra-cellular matrix degradation, and progressive diameter expansion. Additional defects in the anti-inflammatory response may also contribute to AAA progression, however have yet to be characterized robustly. Here, we describe the role of the anti-inflammatory cytokine TSG-6 (TNF-stimulated gene-6) in AAA formation. Methods: Blood and aortic tissue samples were collected from patients undergoing elective AAA screening and open surgical AAA repair. Aortic specimens collected were preserved for IHC or immediately assayed after tissue homogenization. Cytokine concentrations in tissue and plasma were assayed by ELISA. All immune cell populations were assayed using FACS analysis. In vitro, macrophage polarization from monocytes were performed with young, healthy donor PBMCs. Results: TSG-6 was found to be abnormally elevated in both the plasma and aorta of patients with AAA compared to healthy and risk-factor matched non-AAA donors. We observed the highest tissue concentration of TSG-6 in the less diseased proximal and distal shoulders compared to the central aspect of the aneurysm. IHC localized the majority of TSG-6 to the tunica media with minor expression in the tunica adventitia of the aortic wall. Higher concentrations of both M1 and M2 macrophages where also observed in the aortic wall, however M1/M2 ratios were unchanged from healthy controls. Additionally, we observed no difference in M1/M2 ratios in the peripheral blood of risk-factor matched non-AAA and AAA patients. Interesting, TSG-6 inhibited the polarization of the anti-inflammatory M2 phenotype in vitro . Conclusions: AAA formation results from an imbalance of inflammatory forces causing aortic wall infiltration of mononuclear cells leading to resultant vessel breakdown. From our results, we suggest TSG-6 is elevated in the AAA patient as a compensatory anti-inflammatory feedback mechanism. However, it’s effects may be abrogated by defects in CD44, its cognate receptor or downstream signaling pathways, future areas for investigation.