Interpreting permeability as a function of free drug fraction: The case studies of cyclodextrins and liposomes.

In order to solubilize poorly soluble active pharmaceutical ingredients, various strategies have been implemented over the years, including the use of nanocarriers, such as cyclodextrins and liposomes. However, improving a drug's apparent solubility does not always translate to enhanced bioavailability. This work aimed to investigate to which extent complexation with cyclodextrins and incorporation into liposomes influence drug in vitro permeability and to find a mechanistic description of the permeation process. For this purpose, we investigated hydroxypropyl-ß-cyclodextrin (HP-ß-CD) and phosphatidylcholine liposomes formulations of three chemically diverse compounds (atenolol, ketoprofen and hydrocortisone). We studied drug diffusion of the formulations by UV-localized spectroscopy and advanced data fitting to extract parameters such as diffusivity and bound-/free drug fractions. We then correlated this information with in vitro drug permeability obtained with the novel PermeaPadⓇ barrier. The results showed that increased concentration of HP-ß-CD leads to increased solubilization of the poorly soluble unionized ketoprofen, as well as hydrocortisone. However, this net increment of apparent solubility was not proportional to the increased flux measured. On the other hand, normalising the flux over the empirical free drug concentration, i.e., the free fraction, gave a meaningful absolute permeability coefficient. The results achieved for the liposomal formulation were consistent with the finding on cyclodextrins. In conclusion, we proved the adequacy and usefulness of our method for calculating free drug fractions in the examined enabling formulations, supporting the validity of the established drug diffusion/permeation theory that the unbounded drug fraction is the main driver for drug permeation across a membrane.

Reversible Immunosensor for the Continuous Monitoring of Cortisol in Blood Plasma Sampled with Microdialysis.

Cortisol is a steroid hormone involved in a wide range of medical conditions. The level of the hormone fluctuates over time, but with traditional laboratory-based assays, such dynamics cannot be monitored in real time. Here, a reversible cortisol sensor is reported that allows continuous monitoring of cortisol in blood plasma using sampling by microdialysis. The sensor is based on measuring single-molecule binding and unbinding events of tethered particles. The particles are functionalized with antibodies and the substrate with cortisol-analogues, causing binding and unbinding events to occur between particles and substrate. The frequency of binding events is reduced when cortisol is present in the solution as it blocks the binding sites of the antibodies. The sensor responds to cortisol in the high nanomolar to low micromolar range and can monitor cortisol concentrations over multiple hours. Results are shown for cortisol monitoring in filtered and in microdialysis-sampled human blood plasma.

NR3C1 gene methylation and cortisol levels in preterm and healthy full-term infants in the first 3 months of life.

Aim: To describe NR3C1 exon-1F methylation and cortisol levels in newborns. Materials & methods: Preterm ≤1500 g and full-term infants were included. Samples were collected at birth and at days 5, 30 and 90 (or at discharge). Results: 46 preterm and 49 full-term infants were included. Methylation was stable over time in full-term infants (p = 0.3116) but decreased in preterm infants (p = 0.0241). Preterm infants had higher cortisol levels on the fifth day, while full-term infants showed increasing levels (p = 0.0177) over time. Conclusion: Hypermethylated sites in NR3C1 at birth and higher cortisol levels on day 5 suggest that prematurity, reflecting prenatal stress, affects the epigenome. Methylation decrease over time in preterm infants suggests that postnatal factors may modify the epigenome, but their role needs to be clarified.

We investigated the methylation of a gene, NR3C1 exon-1F, and cortisol levels in newborns. DNA methylation is a biochemical process that can modify gene activity. In the case of this gene, higher methylation might be associated with higher cortisol levels. We studied 46 preterm infants (born weighing 1500 g or less) and 49 full-term infants. Our results revealed that the preterm infants had hypermethylation at birth and higher cortisol levels on day 5, but decreasing methylation and stable cortisol levels over time. Meanwhile, methylation remained stable and cortisol levels increased in full-term babies with time. These unexpected results suggest that prematurity can be associated with prenatal epigenetic changes in the NR3C1 gene, but postnatal factors may induce further modifications. More research is needed to understand these findings better.

In silico formulation prediction of drug/cyclodextrin/polymer ternary complexes by machine learning and molecular modeling techniques.

Ternary cyclodextrin (CD) complexes (drug/CD/polymer) can effectively improve the solubility of water-insoluble drugs with large size than binary CD formulations. However, ternary formulations are screened by a trial-and-error approach, which is laborious and material-wasting. Current research aims to develop a prediction model for ternary CD formulations by combined machine learning and molecular modeling. 596 ternary formulations data were collected to build a prediction model by machine learning. The random forest model achieved good performance with R2 = 0.887 in ST prediction and R2 = 0.815 in ST/SB prediction. Two ternary formulations (Hydrocortisone/ß-CD/HPMC and dovitinib/γ-CD/CMC) were used to validate the prediction model. Molecular modeling results showed that HPMC not only warped around hydrocortisone but also prevented CD molecules from self-aggregation to increase solubility. In conclusion, a prediction model for the ternary CD formulations was successfully developed, which will significantly accelerate the formulation screening process to benefit the formulation development of water-insoluble drugs.

Summated training and match load predictors of salivary immunoglobulin-A, alpha-amylase, testosterone, cortisol and T:C profile changes in elite-level professional football players: A longitudinal analysis.

We examined how summated training and match load measures relate to salivary immunological and hormonal profile changes in professional football players. Data were collected from 18 elite-level professional male football players from one English Championship team across a complete 40 wk competitive season. Daily training (micro-technology) and match (computerised tracking) measures of total, high-speed and high-metabolic load running distance and sprint, acceleration, deceleration and sRPE load were converted into exponentially weighted moving average "acute" (7d), "chronic" (28d) and acute:chronic composite load measures. Bi-weekly morning saliva samples were analysed for immunoglobulin-A, alpha-amylase, testosterone, cortisol and testosterone:cortisol. A two-stage data reduction technique using partial least squares modelling and a backward stepwise selection procedure determined the most parsimonious model for each salivary variable. Testosterone had non-linear relationships with chronic total (P = 0.015; Cohen's D: large), high-metabolic load (P = 0.001;small) and high-speed (P = 0.001;trivial) running distance and linear relationships with chronic sRPE (P = 0.002;moderate ↓) and acute:chronic high-speed running distance (P = 0.001; trivial ↑). Cortisol had a non-linear relationship with chronic high-speed running distance (P = 0.001;trivial). Testosterone:cortisol had non-linear relationships with chronic decelerations (P = 0.039;small) and chronic summated acceleration and deceleration load (P = 0.039;small). Non-linear relationships typically indicated optimal hormonal responses at squad mean loads. No load variables clearly related to salivary immunoglobulin-A or alpha-amylase changes. We conclude that chronic total and high-intensity load measures relate to hormonal changes and might be useful indicators of player readiness. Acute load variables were not related to immunological or hormonal changes and consequently, should not be used as surrogate measures of player readiness in isolation.

Salivary cortisone as potential predictor of occupational exposure to noise and related stress.

Salivary cortisone strongly correlates with serum cortisol, and since it is less invasive to measure salivary cortisone than serum cortisol and easier than to measure cortisol in saliva, as its concentrations are much lower, we wanted to compare salivary cortisone and cortisol levels as markers of noise-induced stress reaction. The study included 104 participants aged 19-30 years, 50 of whom were exposed to occupational noise ≥85 dB(A) and 54 non-exposed, control students. All participants took samples of their saliva with Salivette® Cortisol synthetic swabs on three consecutive working days first thing in the morning. Salivary cortisone and cortisol levels were determined with high-performance liquid chromatography. In addition, they completed a 10-item Perceived Stress Scale (PSS-10) questionnaire, and occupationally noise-exposed participants also completed the Health and Safety Executive (HSE) questionnaire on occupational psychosocial risks. The exposed participants had significantly higher cortisone (P<0.001) and cortisol (P<0.001) levels than controls, and the correlation between cortisone and cortisol levels in the exposed participants was strong (ϱ =0.692, P<0.001), which suggests that salivary cortisone can replace cortisol measurements in saliva as a more reliable method than salivary cortisol and less invasive than serum cortisol. However, the level of perceived stress scored on PSS-10 in the exposed participants did not differ significantly from stress reported by controls, but correlated negatively with cortisone levels, which is contrary to our expectations and raises questions as to why.

Salivary cortisol and eye temperature changes during endurance competitions.

BACKGROUND: The purpose of this study was to investigate the usefulness of salivary cortisol (SC) and eye temperature measured by infrared thermography (IRTET) as biomarkers to manage competitions more effectively and monitor horse welfare in endurance competitions. Based on previous studies, it was hypothesised that pre-exercise baseline SC and IRTET would be higher in younger or less experienced horses, and that post-exercise variation from baseline would be higher in the top finishers. RESULTS: Salivary cortisol measured in 61 competing at qualifier 40 km and 80 km rides showed an abrupt variation (93-256% rise) of the baseline SC levels [median ± interquartile range (IQR) = 0.27 ng/dl ± 0.36] obtained at the Pre-Inspection (PI) into Vet Gate (VG)1 independently of the covered distance, but modest or even lower in the subsequent Vet Gates, e.g. VG2 or VG3. The IRTET measured concomitantly in 16 horses showed significant (p < 0.05) higher levels at the PI in less experienced horses participating in the 40 km ride (median ± IQR = 35.7 °C ± 1.4) than their counterparts in the 80 km ride (median ± IQR = 35.0 °C ± 1.5), but not SC. Baseline SC levels at the PI of horses classifying in the Top5 in the 40 km ride category were significantly (p < 0.05) higher median ± IQR = 0.90 ng/ml ±0.61) when compared to horses positioned from 10th position on (median ± IQR = 0.16 ng/ml ±0.40). A lower IRTET in the PI was correlated with better placement (p < 0.05) and those in the Top5 (median ± IQR = 33.9 °C ± 0.0) had a significantly (p < 0.5) higher variation (+ 10.65%) into the last VG. CONCLUSION: Pre-exercise baseline IRTET levels, but not SC, were higher in less experienced horses in the 40 compared to their counterparts in the 80 km ride competitions. SC and IRTET showed different indications according to the competition. In the40 km ride competition, higher baseline pre-exercise SC levels seemed to be linked to a better classification outcome. In contrast, in the 80 km ride horses, the higher IRTET variation from pre-exercise into final Vet Gate was the parameter associated with a better performance. A more controlled environment and a larger sample are needed to confirm these results and monitor horse welfare in competitions.

Novel 2-(Adamantan-1-ylamino)Thiazol-4(5H)-One Derivatives and Their Inhibitory Activity towards 11ß-HSD1-Synthesis, Molecular Docking and In Vitro Studies.

A common mechanism in which glucocorticoids participate is suggested in the pathogenesis of such metabolic diseases as obesity, metabolic syndrome, or Cushing's syndrome. The enzyme involved in the control of the availability of cortisol, the active form of the glucocorticoid for the glucocorticoid receptor, is 11ß-HSD1. Inhibition of 11ß-HSD1 activity may bring beneficial results for the alleviation of the course of metabolic diseases such as metabolic syndrome, Cushing's syndrome or type 2 diabetes. In this work, we obtained 10 novel 2-(adamantan-1-ylamino)thiazol-4(5H)-one derivatives containing different substituents at C-5 of thiazole ring and tested their activity towards inhibition of two 11ß-HSD isoforms. For most of them, over 50% inhibition of 11ß-HSD1 and less than 45% inhibition of 11ß-HSD2 activity at the concentration of 10 µM was observed. The binding energies found during docking simulations for 11ß-HSD1 correctly reproduced the experimental IC50 values for analyzed compounds. The most active compound 2-(adamantan-1-ylamino)-1-thia-3-azaspiro[4.5]dec-2-en-4-one (3i) inhibits the activity of isoform 1 by 82.82%. This value is comparable to the known inhibitor-carbenoxolone. The IC50 value is twice the value determined by us for carbenoxolone, however inhibition of the enzyme isoform 2 to a lesser extent makes it an excellent material for further tests.

Highly Sensitive Colorimetric Assay of Cortisol Using Cortisol Antibody and Aptamer Sandwich Assay.

In this study, cortisol, which is a key stress hormone, could be detected sensitively via the colorimetric assay of a polycarbonate (PC) and glass substrate by the sandwich assay of cortisol monoclonal antibody (c-Mab) and cortisol specific binding aptamer (c-SBA). A highly sensitive change in colorimetry with a limit of detection (LOD) of cortisol of 100 fM could be attained on the optically transparent substrate using the antibody aptamer sandwich (AAS) assay by corresponding stacks of 5 nm gold nanoparticles (Au NPs). The Au NPs were conjugated by the c-SBA and the c-Mab was tethered on the PC and glass substrates. For the AAS method, a simple UV-Vis spectrophotometer was adopted to quantify the cortisol concentrations at an absorbance wavelength of 520 nm. Therefore, this study demonstrates the versatility of the AAS method to measure very low concentrations of cortisol in diagnostic applications.

Prediction of salivary cortisol level by electroencephalography features.

Change in cortisol affects brain EEG signals. So, the identification of the significant EEG features which are sensitized to cortisol concentration was the aim of the present study. From 468 participated healthy subjects, the salivary samples were taken to test the cortisol concentration and EEG signal recording was done simultaneously. Then, the subjects were categorized into three classes based on the salivary cortisol concentration (<5, 5-15 and >15 nmol/l). Some linear and nonlinear features extracted and finally, in order to investigate the relationship between cortisol level and EEG features, the following steps were taken on features in sequence: Genetic Algorithm, Neighboring Component Analysis, polyfit, artificial neural network and support vector machine classification. Two classifications were considered as following: state 1 categorized the subjects into three groups (three classes) and the second state put them into two groups (group 1: class 1 and 3, group 2: class 2). The best classification was done using ANN in the second state with the accuracy=94.1% while it was 92.7% in the first state. EEG features carefully predicted the cortisol level. This result is applicable to design the intelligence brain computer machines to control stress and brain performance.

Cyclic Cushing's Syndrome - A Diagnostic Challenge.

Cyclic Cushing's syndrome (also known as intermittent or periodic) is a disease characterized by periods of transient hypercortisolemia shifting into periods of normo- and/or hypocortisolemia. Diagnosis of cyclic Cushing's syndrome is based on at least three periods of confirmed hypercortisolemia interspersed by two periods of normocortisolemia. Cyclic Cushing's syndrome is one of the greatest challenges in modern endocrinology due to its diverse clinical picture, unpredictable duration and frequency of phases, and various etiologies. We discuss a diagnostic algorithm for periodic hypercortisolemia with special regard to hair cortisol analysis and desmopressin stimulation test which both seem to be helpful in finding the correct answer.

Early Metabolic Benefits of Switching Hydrocortisone to Modified Release Hydrocortisone in Adult Adrenal Insufficiency.

Purpose: To compare metabolic effects of modified release hydrocortisone (MR-HC) with standard hydrocortisone (HC) therapies in adults with Adrenal Insufficiency (AI). Methods: Adult patients (n = 12) with AI, established on HC therapy, were recruited from Endocrinology clinics at University Hospitals Coventry and Warwickshire (UHCW), UK. Baseline (HC) metabolic assessments included fasting serum HbA1C, lipid and thyroid profiles, accurate measures of body composition (BodPod), and 24-h continuous measures of energy expenditure including Sleeping Metabolic Rate (SMR) using indirect calorimetry within the Human Metabolism Research Unit, UHCW. All participants then switched HC to MR-HC with repeat (MR-HC) metabolic assessments at 3 months. Paired-sample t-tests were used for data comparisons between HC and MR-HC assessments: P-value <0.05 was considered significant. Results: Following exclusion of 2 participants, analyses were based on 10 participants. Compared with baseline HC data, following 3 months of MR-HC therapy mean fat mass reduced significantly by -3.2 kg (95% CI: -6.0 to -0.4). Mean (SD) baseline HC fat mass vs repeat MR-HC fat mass: 31.9 kg (15.2) vs 28.7 kg (12.8) respectively, P = 0.03. Mean SMR increased significantly by +77 kcal/24 h (95% CI: 10-146). Mean (SD) baseline HC SMR vs repeat MR-HC SMR: 1,517 kcal/24 h (301) vs 1,594 kcal/24 h (344) respectively, P = 0.03. Mean body fat percentage reduced significantly by -3.4% (95% CI: -6.5 to -0.2). Other measures of body composition, energy expenditure, and biochemical analytes were equivalent between HC and MR-HC assessments. Conclusions: In adults with AI, switching from standard HC to MR-HC associates with early metabolic benefits of reduced fat mass and increased SMR.

Modified-Release Hydrocortisone in Congenital Adrenal Hyperplasia.

CONTEXT: Standard glucocorticoid therapy in congenital adrenal hyperplasia (CAH) regularly fails to control androgen excess, causing glucocorticoid overexposure and poor health outcomes. OBJECTIVE: We investigated whether modified-release hydrocortisone (MR-HC), which mimics physiologic cortisol secretion, could improve disease control. METHODS: A 6-month, randomized, phase 3 study was conducted of MR-HC vs standard glucocorticoid, followed by a single-arm MR-HC extension study. Primary outcomes were change in 24-hour SD score (SDS) of androgen precursor 17-hydroxyprogesterone (17OHP) for phase 3, and efficacy, safety and tolerability of MR-HC for the extension study. RESULTS: The phase 3 study recruited 122 adult CAH patients. Although the study failed its primary outcome at 6 months, there was evidence of better biochemical control on MR-HC, with lower 17OHP SDS at 4 (P = .007) and 12 (P = .019) weeks, and between 07:00h to 15:00h (P = .044) at 6 months. The percentage of patients with controlled 09:00h serum 17OHP (< 1200 ng/dL) was 52% at baseline, at 6 months 91% for MR-HC and 71% for standard therapy (P = .002), and 80% for MR-HC at 18 months' extension. The median daily hydrocortisone dose was 25 mg at baseline, at 6 months 31 mg for standard therapy, and 30 mg for MR-HC, and after 18 months 20 mg MR-HC. Three adrenal crises occurred in phase 3, none on MR-HC and 4 in the extension study. MR-HC resulted in patient-reported benefit including menses restoration in 8 patients (1 on standard therapy), and 3 patient and 4 partner pregnancies (none on standard therapy). CONCLUSION: MR-HC improved biochemical disease control in adults with reduction in steroid dose over time and patient-reported benefit.

Cytotoxic and genotoxic effects in human gingival fibroblast and ions release of endodontic sealers incorporated with nanostructured silver vanadate.

The cytotoxic and genotoxic effects of commercial endodontic sealers (AH Plus, Sealer 26 and Endomethasone N) incorporated with nanostructured silver vanadate decorated with silver nanoparticles (AgVO3 - at concentrations 2.5, 5, and 10%) on human gingival fibroblast (HGF), and the silver (Ag+ ) and vanadium (V4+ /V5+ ) ions release were evaluated. Cytotoxicity, cell death, and genotoxicity tests were carried out with extract samples of 24-hr and 7-days. The release of Ag+ and V4+ /V5+ was evaluated. Cytotoxicity in HGF was caused by AH Plus (AP) with 5 and 10% of AgVO3 (83.84 and 67.49% cell viability, respectively) with 24-hr extract (p < 0.05), as well as all concentrations of AP with 7-days extract (p < 0.05 -AP 0% = 73.17%; AP 2.5% = 75.07%; AP 5% = 70.62%; AP 10% = 68.46% cell viability). The commercial sealers Sealer 26 (S26) and Endomethasone N (EN) were cytotoxic (p < 0.05 - S26 0% = 34.81%; EN 0% = 20.99% cell viability with 7-days extract). AP 10% with 7-days extract induced 32% apoptotic cells in HGF (p < 0.05). Genotoxic effect was not observed. The AP groups released more Ag+ , while S26 and EN released more V4+ /V5+ in 24 hr. The Ag+ can be cytotoxic. In conclusion, the cytotoxicity caused to HGF can be attributed by the commercial sealers and enhanced by incorporation of AgVO3 , was not observed genotoxic effect, and apoptosis was induced only by AH Plus 10% 7-days extract. Ag+ can influence cell viability.

Gut feelings about bacterial steroid-17,20-desmolase.

Advances in technology are only beginning to reveal the complex interactions between hosts and their resident microbiota that have co-evolved over centuries. In this review, we present compelling evidence that implicates the host-associated microbiome in the generation of 11ß-hydroxyandrostenedione, leading to the formation of potent 11-oxy-androgens. Microbial steroid-17,20-desmolase cleaves the side-chain of glucocorticoids (GC), including cortisol (and its derivatives of cortisone, 5α-dihydrocortisol, and also (allo)- 3α, 5α-tetrahydrocortisol, but not 3α-5ß-tetrahydrocortisol) and drugs (prednisone and dexamethasone). In addition to side-chain cleavage, we discuss the gut microbiome's robust potential to transform a myriad of steroids, mirroring much of the host's metabolism. We also explore the overlooked role of intestinal steroidogenesis and efflux pumps as a potential route for GC transport into the gut. Lastly, we propose several health implications from microbial steroid-17,20-desmolase function, including aberrant mineralocorticoid, GC, and androgen receptor signaling in colonocytes, immune cells, and prostate cells, which may exacerbate disease states.

Regulation of the early expression of MAFbx/atrogin-1 and MuRF1 through membrane-initiated cortisol action in the skeletal muscle of rainbow trout.

Glucocorticoids are key stress-related hormones in vertebrates, with cortisol being the main glucocorticoid in teleosts. Glucocorticoids exert their effects through two mechanisms of action: genomic/classic and membrane initiated. In mammals, cortisol-mediated stress has been found to be associated with increased expression of critical atrophy-related genes (atrogenes), such as MAFbx/atrogin-1 and murf1/trim63. However, the direct impact of cortisol on the early regulation of atrogene expression in teleost skeletal muscle and the contribution of membrane-initiated cortisol action to this process have not been identified. In this work, the mRNA levels of atrogin-1 and murf1 were assessed in isolated myotubes and skeletal muscle of rainbow trout administered with cortisol or cortisol-BSA. This latter compound is a membrane-impermeable cortisol analog that exclusively induces membrane-initiated effects. We found that cortisol (10 mg/kg) first decreased the expression of both atrogenes at 3 h of treatment and then increased their expression at 9 h of treatment in the skeletal muscle of rainbow trout. Additionally, the in vitro analysis suggested that membrane-initiated cortisol action regulates murf1 but not atrogin-1 in rainbow trout myotubes. Using RU486 to selectively block glucocorticoid receptor (GR), we found that early downregulation of murf1 is potentially mediated by membrane GR signaling in myotubes. Considering the results of both the in vivo and in vitro approaches, we suggest that membrane-initiated cortisol action regulates the early expression of atrophy-related processes in teleosts.

Impedimetric determination of cortisol using screen-printed electrode with aptamer-modified magnetic beads.

A non-invasive aptamer-based electrochemical biosensor using disposable screen-printed graphene electrodes (SPGEs) was developed for simple, rapid, and sensitive determination of cortisol levels. Selective detection of cortisol based on a label-free electrochemical assay was achieved by specific recognition of the cortisol DNA aptamer (CApt). The CApt was modified with streptavidin magnetic beads (MBs) before simple immobilization onto the electrode surface using a neodymium magnet. The electrochemical behavior of the aptamer-based biosensor was assessed by using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) (vs Ag/AgCl). The specific binding between cortisol and CApt resulted in a decrease in charge transfer resistance (Rct) from EIS using [Fe(CN)6]3-/4- with increasing cortisol concentration. Under optimal conditions, a linear range from 0.10 to 100 ng/mL with a low detection limit (3SD/slope) of 2.1 pg/mL was obtained. Furthermore, the proposed biosensing system exhibited a satisfactory recovery in the range 97.4-109.2% with 5.7-6.6% RSD in spiked artificial human sweat. Regarding the applications of this tool, the aptamer-based biosensor has potential to be a versatile and point-of-care (POC) device for simple, sensitive, selective, disposable, and low-cost cortisol detection.

A randomized cross-over trial assessing salivary and urinary cortisol concentrations after alfaxalone and propofol administration in healthy cats.

The aim of this study was to assess the effects of commonly used anaesthetics alfaxalone and propofol on salivary and urinary cortisol in healthy cats. Fifteen male castrated research-purposed cats received randomly intravenous continuous rate infusions of 8 mg/kg/h of alfaxalone, 12 mg/kg/h of propofol and 2 ml/kg/h of Lactated Ringer's solution for 30 min, with intervals of 6 days between treatments. Saliva samples were collected for 24 h before each infusion and for 24 h from the start of each infusion. Urine was collected as single pooled samples over each 24 h period. Mean integrated saliva cortisol responses in cats treated with alfaxalone were greater than responses of cats treated with propofol (P = 0.034) and controls (P = 0.017). Integrated responses in cats treated with propofol did not differ from controls. The mean urinary cortisol/creatinine ratio (UCCR) was higher on the day of treatment than the day before treatment in cats treated with alfaxalone (P < 0.0001) and in cats treated with propofol (P = 0.0168) and did not differ between days in cats treated with lactated Ringer's solution. The mean UCCR was higher in cats treated with alfaxalone than in cats treated with lactated Ringer's solution (P = 0.0020) on the day of treatment. Mean total urinary cortisol over 24 h was greater in cats treated with alfaxalone than controls (P = 0.0267). In conclusion, alfaxalone increased short-term salivary and urinary cortisol concentrations in healthy cats as compared to propofol and a control group of non-anesthetised cats.

Beta-Cyclodextrin-Decorated Magnetic Activated Carbon as a Sorbent for Extraction and Enrichment of Steroid Hormones (Estrone, ß-Estradiol, Hydrocortisone and Progesterone) for Liquid Chromatographic Analysis.

A ß-cyclodextrin-decorated magnetic activated carbon adsorbent was prepared and characterized using various analytical techniques (X-ray diffraction (XRD), scanning electron microscopy-electron diffraction spectroscopy (SEM-EDS) and transmission electron microscopy (TEM)), and the adsorbent was used in the development of a magnetic solid-phase microextraction (MSPE) method for the preconcentration of estrone, ß-estradiol, hydrocortisone and progesterone in wastewater and river water samples. This method was optimized using the central composite design in order to determine the experimental parameters affecting the extraction procedure. The quantification of hormones was achieved using high-performance liquid chromatography equipped with a photodiode array detector (HPLC-DAD). Under optimum conditions, the linearity ranged from 0.04 to 300 µg L-1 with a correlation of determinations of 0.9969-0.9991. The limits of detection and quantification were between 0.01-0.03 and 0.033-0.1 µg L-1, with intraday and interday precisions at 1.1-3.4 and 3.2-4.2. The equilibrium data were best described by the Langmuir isotherm model, and high adsorption capacities (217-294 mg g-1) were obtained. The developed procedure demonstrated high potential as an effective technique for use in wastewater samples without significant interferences, and the adsorbent could be reused up to eight times.

Hair cortisol in mother-child dyads: examining the roles of maternal parenting and stress in the context of early childhood adversity.

Physiological stress is thought to be one way that early adversity may impact children's health. How this occurs may be related to parental factors such as mothers' own stress and parenting behaviour. Hair cortisol offers a novel method for examining long-term physiological stress in mother-child dyads. The current study used hair cortisol to examine the role that maternal physiological stress and parenting behaviours play in explaining any effects of adversity on young children's physiological stress. This cross-sectional study comprised 603 mother-child dyads at child age 2 years, recruited during pregnancy for their experience of adversity through an Australian nurse home visiting trial. Hair cortisol data were available for 438 participating mothers (73%) and 319 (53%) children. Confirmatory factor analysis was used to define composite exposures of economic (e.g. unemployment, financial hardship) and psychosocial (e.g. poor mental health, family violence) adversity, and positive maternal parenting behaviour (e.g. warm, responsive). Structural equation modelling examined maternal mediating pathways through which adversity was associated with children's physiological stress. Results of the structural model showed that higher maternal and child physiological stress (hair cortisol) were positively associated with one another. Parenting behaviour was not associated with children's physiological stress. There was no evidence of any mediating pathways by which economic or psychosocial adversity were associated with children's physiological stress. The independent association identified between maternal and child hair cortisol suggests that young children's physiological stress may not be determined by exogenous environmental exposures; endogenous genetic factors may play a greater role.