Hormones,behavior, and social network analysis: Exploring associations between cortisol,testosterone, and network structure

We used a new interdisciplinary paradigm of social network analysis (SNA) to investigate associations between hormones and social network structures. We examine these biobehavioral processes and test hypotheses about how hormones are associated with social network structures using exponential random graph modeling (ERGM) in a cohort of first-year students (n = 74; 93% female; M age = 27 years) from a highly competitive, accelerated nursing program. Participants completed friendship nominations and as a group simultaneously donated saliva (later assayed for cortisol and testosterone). ERGM analyses revealed that salivary cortisol levels were inversely associated with the number of outgoing ties (i.e., network activity). By contrast, testosterone was not related to friendship network structure. Integration of SNA and salivary bioscience creates a novel approach to understanding hormone–behavior relationships within the context of human social ecologies.     

High self-perceived stress and many stressors, but normal diurnal cortisol rhythm, in adults with ADHD (attention-deficit/hyperactivity disorder)

Attention-deficit/hyperactivity disorder (ADHD) in adults is associated with significant impairment in many life activities and may thus increase the risk of chronic stress in everyday life. We compared adults with a DSM-IV ADHD diagnosis (n = 28) with healthy controls (n = 28) regarding subjective stress and amounts of stressors in everyday life, diurnal salivary cortisol in the everyday environment and salivary cortisol before and after cognitive stress in a laboratory setting. The association between cortisol concentrations and impulsivity was also investigated. Consistent with assumptions, individuals with ADHD reported significantly more self-perceived stress than controls, and subjective stress correlated with the amount of stressors in everyday life. The two groups were comparable with respect to overall diurnal cortisol levels and rhythm, as well as in pre- and post-stress cortisol concentrations. Post-stress cortisol (but not baseline cortisol) concentration was positively correlated with impulsivity. The group with high post-stress cortisol also reported more symptoms of depression and anxiety, as well as self-perceived stress and stressors in every-day life. The diagnosis of ADHD significantly increased the risk of belonging to the group with high post-stress cortisol levels. The results in this study warrant a focus not only on the primary diagnosis of ADHD, but also calls for a broader assessment of stressors and subjective stress in everyday life, as well as support comprising stress management and coping skills.     

Cortisol release and heart rate variability in horses during road transport

Based on plasma cortisol concentrations it is widely accepted that transport is stressful to horses. So far, cortisol release during transport has not been evaluated in depth by non-invasive techniques such as analysis of salivary cortisol and faecal cortisol metabolites. Transport also causes changes in heart rate and heart rate variability (HRV). In this study, salivary cortisol, faecal cortisol metabolites, heart rate and HRV in horses transported by road for short (one and 3.5 h) and medium duration (8 h) were determined. With the onset of transport, salivary cortisol increased immediately but highest concentrations were measured towards the end of transport (4.1 ± 1.6, 4.5 ± 2.6, 6.5 ± 1.8 ng/ml in horses transported for one, 3.5 and 8 h, respectively). Faecal cortisol metabolite concentrations did not change during transport, but 1 day after transport for 3.5 and 8 h had increased significantly (p < 0.01), reflecting intestinal passage time. Compared to salivary cortisol, changes in faecal cortisol metabolites were less pronounced. Heart rate increased and beat-to-beat (RR) interval decreased (p < 0.05) with the onset of transport. Standard deviation of heart rate increased while root mean square of successive RR differences (RMSSD) decreased in horses transported for 3.5 (from 74 ± 5 to 45 ± 6 ms) and 8 h (from 89.7 ± 7 to 59 ± 7 ms), indicating a reduction in vagal tone. In conclusion, transport of horses over short and medium distances leads to increased cortisol release and changes in heart rate and HRV indicative of stress. The degree of these changes is related to the duration of transport. Salivary cortisol is a sensitive parameter to detect transient changes in cortisol release.     

The effects of saliva collection,handling and storage on salivary testosterone measurement

Several endocrine parameters commonly measured in plasma, such as steroid hormones, can be measured in the oral fluid. However, there are several technical aspects of saliva sampling and processing that can potentially bias the validity of salivary testosterone measurement. The aim of this study was to evaluate the effects caused by repeated sampling; 5 min centrifugation (at 2000, 6000 or 10,000g); the stimulation of saliva flow by a cotton swab soaked in 2% citric acid touching the tongue; different storage times and conditions as well as the impact of blood contamination on salivary testosterone concentration measured using a commercially available ELISA kit. Fresh, unprocessed, unstimulated saliva samples served as a control. Salivary testosterone concentrations were influenced neither by repeated sampling nor by stimulation of salivary flow. Testosterone levels determined in samples stored in various laboratory conditions for time periods up to 1 month did not differ in comparison with controls. For both genders, salivary testosterone levels were substantially reduced after centrifugation (men F = 29.1; women F = 56.17, p < 0.0001). Blood contamination decreased salivary testosterone levels in a dose-dependent manner (men F = 6.54, p < 0.01, F = 5.01, p < 0.05). Salivary testosterone can be considered A robust and stable marker. However, saliva processing and blood leakage can introduce bias into measurements of salivary testoterone using ELISA. Our observations should be considered in studies focusing on salivary testosterone.     

Abolished circadian rhythm of salivary cortisol in elite artistic gymnasts

Objective

The aim of this study was to evaluate the effects of intensive physical exercise and acute psychological stress during high level athletic competition as reflected on the levels of salivary cortisol in elite artistic gymnasts (AGs).

Design

The study included 239 AGs (142 females-97 males) who participated in the European Championship of Gymnastics in 2006 and 81 adolescents (40 females-41 males), matched for age, as controls. All athletes participated voluntarily in all or parts of the study, providing samples or data for each of the variables measured. Height, weight, body fat, lean body mass (LBM), bone age and Tanner stage of puberty were assessed and data concerning the time of thelarche, adrenarche and menarche as well as, the onset and the intensity (hours per week) of training were obtained.

Methods

Saliva samples were collected, the morning before training and in the afternoon shortly after the competition. From controls, the saliva samples were collected in the morning. Cortisol concentrations were measured using a chemiluminescence method. Acute stress was assessed using a questionnaire designed for the study.

Results

No difference was found between morning and afternoon salivary cortisol levels in both male and female AGs (females: AM: 15.45 ± 7.45 nmol/l vs PM: 15.73 ± 9.38 nmol/l; males: AM: 10.21 ± 5.52 nmol/l vs PM: 9.93 ± 13.8 nmol/l, p > 0.05).Female AGs presented higher levels of morning salivary cortisol than female controls (p < 0.05). Both male and female AGs had higher degree of psychological stress in comparison with controls (p < 0.001, p < 0.013, respectively). Female AGs had higher morning and afternoon salivary cortisol levels (p < 0.01, p < 0.01, respectively) and higher degree of stress (p < 0.003) than males.

Conclusions

In elite AGs the diurnal rhythm of salivary cortisol has been abolished, probably due to the strenuous training and competition conditions. Female AGs presented higher levels of morning salivary cortisol and psychological stress compared to both male AGs and female controls. The long term consequences of these modifications of the HPA axis remain to be elucidated.     

Negative relationships in the family-of-origin predict attenuated cortisol in emerging adults

Negative childhood family environments have been associated with stress-related physical and psychological health consequences across the lifespan. The present study examined the relation between adverse relationships in the family of origin and physiological stress response, as measured by salivary cortisol, in emerging adulthood. Seventy-six university students (age range = 18-22) selected from intact married families-of-origin characterized by either negative (n = 39) or positive (n = 37) relationship quality engaged in a challenging role play task. Results from multilevel models indicated that those from negative families exhibited significantly lower salivary cortisol across the task than those from positive families. This relation did not change in strength or direction after controlling for experiences with abuse or recent anxiety or depressive symptoms. These findings suggest the significance of early family relationships on the long-term activity of the HPA axis.     

A re-investigation of saliva collection procedures that highlights the risk of potential positive interference in cortisol immunoassay

Background

The use of saliva for measurement of cortisol permits non-invasive study of adrenal function, but collection can be technically difficult, particularly in small infants. Saliva collection can be assisted by citric acid to increase saliva flow, or by the use of cotton or polyester swabs in the mouth.

Aim

To determine whether different methods of saliva collection affect cortisol radioimmunoassay (RIA) performance.

Experimental

Cortisol was measured in saliva collected from 16 adults using intra-oral cotton swabs or polyester swabs, compared with saliva dribbled directly into a pot either alone (plain saliva) or after citric acid had been placed on the tongue. An in-house RIA, without prior extraction, was used to measure cortisol with an encapsulated sheep antibody.

Results

Mean (median) salivary cortisol was 10.9 (10.5) nmol L⁻¹ in plain saliva, 10.4 (8.4) nmol L⁻¹ in citric acid stimulated saliva; 25.3 (25.1) nmol L⁻¹ in saliva collected on cotton swabs, and 27.9 (27.3) nmol L⁻¹ collected on polyester swabs. Cortisol in saliva collected using citric acid was not significantly different from plain saliva (p = 0.997), but cortisol in saliva collected using cotton and polyester swabs was significantly higher than that of plain saliva (p < 0.01).

Conclusion

The use of cotton or polyester swabs for collection of saliva can result in spuriously high levels of cortisol when measured by RIA.     

Measurement of cortisol,cortisone, prednisolone,dexamethasone and 11-deoxycortisol with ultra high performance liquid chromatography–tandem mass spectrometry: Application for plasma,plasma ultrafiltrate,urine and saliva in a routine laboratory

We describe an ultra high performance liquid chromatography–tandem mass spectrometry (UHPLC MS/MS) method suitable for a routine laboratory to determine endogenous and exogenous glucocorticoids in plasma, plasma ultrafiltrate, urine and saliva in a single analytical run. After addition of a multi-analyte internal standard, a standardised sample preparation procedure with solid phase extraction followed, before injecting into a tandem mass spectrometer with positive mode electron spray ionisation and multiple reactant monitoring acquisition. The chromatography time was 3 min. The limit of quantitation for cortisol and cortisone in plasma was 3.75 nmol/L and linearity extended to 2000 nmol/L. The limit of quantitation for cortisol in plasma ultrafiltrate and saliva was 0.6 nmol/L. The limit of quantitation for 11-deoxycortisol and prednisolone was 5 nmol/L and for dexamethasone 1 nmol/L. The intra-assay CV was <5% and the inter-assay CV <10% for all analytes in all matrices. Comparison with an immuno-assay (IA) plasma cortisol method resulted in a regression equation of UHPLC = 0.79 × IA + 31.12 with R² = 0.960 (p < 0.0001). Comparison with a high performance liquid chromatography (HPLC) cortisol method yielded a regression equation of UHPLC = 1.06 × HPLC + 9.82, R² = 0.992 (p < 0.0001). The simultaneous measurement of endogenous and exogenous glucocorticoids contributed to patient care in cases with dexamethasone and metyrapone dynamic tests and unsuspected therapeutic glucocorticoid use.     

In utero cortisol and testosterone exposure and fear reactivity in infancy

Fetal programming is emerging as a major conceptual model for understanding developmental origins of health and disease, including behavioral outcomes. As part of a larger study of prenatal stress and child development, we examined the association between prenatal hormone exposure and fear reactivity, a temperament dimension that is a predictor of long-term behavioral adjustment. Amniotic fluid was collected from a sample of women undergoing clinically indicated amniocentesis for later analysis of cortisol and testosterone. Children with normal birth outcomes were recalled for follow-up assessment at 17 months, at which time we administered an observational assessment of temperament (lab-TAB; n = 108). Information on pregnancy and obstetric outcome was included as covariates. Results indicated that there was a significant association between prenatal testosterone and observed fear reactivity in boys (r(53) = 0.34, p = 0.01); no significant effect was found in girls (r(54) = − 0.07, ns); the effect remained when obstetric, psychosocial, and parental anxiety were controlled for. There was not a significant association between fetal cortisol exposure and fear reactivity. The prediction from in utero testosterone exposure to fear reactivity in boys extends prior research on prenatal testosterone and may represent an association with a general predisposition to greater arousal and reactivity.     

Prenatal exposure to maternal psychosocial stress and HPA axis regulation in young adults

Epidemiological studies have reported associations between measures of size and weight at birth and disease risk in later life. Alteration in the regulation of the hypothalamic-pituitary-adrenal (HPA) axis in response to prenatal stress has been proposed as one underlying mechanism. The present study investigated in humans the association of prenatal psychosocial stress exposure with subsequent HPA axis regulation in adult life, with a focus on measures of response to challenge and feedback sensitivity. Healthy young adults whose mothers experienced severe stress during their pregnancy in form of major negative life events (e.g. death of someone close; prenatal stress (PS) group, n = 31) and an age-matched comparison group (CG, n = 30) underwent the Trier Social Stress Test (TSST) and a 1 μg ACTH_1-24 stimulation test. In addition, a diurnal cortisol profile was assessed. ACTH concentrations following a standardized behavioural challenge paradigm (TSST) were marginally significantly higher in PS subjects than in CG subjects (p = .06). Pre-TSST adrenocortical (cortisol) levels were lower (p = .007), whereas the increase in cortisol in response to the TSST was higher (p = .03) in PS subjects compared to CG subjects. Cortisol concentrations following a pharmacological stimulation test simulating pituitary activity (ACTH_1-24 test) were significantly lower in PS than in CG subjects (p = .006). No differences emerged between the two groups in basal diurnal cortisol levels. This study provides first evidence in humans of an association between prenatal psychosocial stress exposure and subsequent alterations in the regulation of the HPA axis.     

Changes in cortisol release and heart rate and heart rate variability during the initial training of 3-year-old sport horses

Based on cortisol release, a variety of situations to which domestic horses are exposed have been classified as stressors but studies on the stress during equestrian training are limited. In the present study, Warmblood stallions (n = 9) and mares (n = 7) were followed through a 9 respective 12-week initial training program in order to determine potentially stressful training steps. Salivary cortisol concentrations, beat-to-beat (RR) interval and heart rate variability (HRV) were determined. The HRV variables standard deviation of the RR interval (SDRR), RMSSD (root mean square of successive RR differences) and the geometric means standard deviation 1 (SD1) and 2 (SD2) were calculated. Nearly each training unit was associated with an increase in salivary cortisol concentrations (p < 0.01). Cortisol release varied between training units and occasionally was more pronounced in mares than in stallions (p < 0.05). The RR interval decreased slightly in response to lunging before mounting of the rider. A pronounced decrease occurred when the rider was mounting, but before the horse showed physical activity (p < 0.001). The HRV variables SDRR, RMSSD and SD1 decreased in response to training and lowest values were reached during mounting of a rider (p < 0.001). Thereafter RR interval and HRV variables increased again. In contrast, SD2 increased with the beginning of lunging (p < 0.05) and no changes in response to mounting were detectable. In conclusion, initial training is a stressor for horses. The most pronounced reaction occurred in response to mounting by a rider, a situation resembling a potentially lethal threat under natural conditions.     

Psychopathic personality traits and cortisol response to stress: The role of sex, type of stressor, and menstrual phase

Previous research indicates that psychopathic personality traits are associated with lower cortisol secretion in response to stress in men but not in women. The current study explored whether prior null results for women were related to the latency of the cortisol stress response to two different types of stressors. Additionally, accuracy of self-reported menstrual phase was explored via salivary progesterone levels. A mixed-sex sample of 145 participants characterized by high (36 men, 37 women) and low (34 men, 38 women) scores on a screening measure of psychopathic personality traits were randomly assigned to either a performance-based stressor task or a social rejection stressor task. Salivary hormone samples were taken just prior to task onset (baseline) and at 0, 20, 40, and 60 min post-stressor. Results indicated that both men and women characterized by psychopathic personality traits exhibited lower stress-induced cortisol levels to the performance-based task in comparison with controls at 20 min post-stressor. The social rejection task produced a cortisol response 20 min post-stressor in the male controls only. Removal of women with low progesterone from the analyses strengthened the psychopathy group differences. Results could suggest that deficient cortisol production in response to stress might be another important neurobiological feature associated with psychopathic traits, and that biological verification of menstrual phase is an important aspect to obtaining accurate cortisol results in women.     

Seasonal plasticity of the pituitary pars intermedia of the one-humped camel (Camelus dromedarius)

The pituitary pars intermedia of Camelus dromedarius is well developed and completely surrounds the pars nervosa. Two major groups of cells are present: endocrine (ec) and glial-like cells (glc). The ec group is composed of three morphologically distinct cell types. Type I, or polyhedral light cells (LC-I) and type II, or polyhedral dark cells (DC-II), have secretory granules of heterogeneous electron density whose size ranges from 170 to 300 nm. Type III cells are elongated with homogeneous electron-dense secretory granules of 80–200 nm. The glc make up an organized network, form follicles in the centrolobular zones and are positive for vimentin and S-100β immunolabelling. The nerve fibres penetrating the lobe are numerous, and can be classified into two types according to the membrane bound vesicles found in their endings (ne). Ultrastructural quantitative analysis revealed significant variations in PI elements between winter and summer seasons (F = 8.014, p = 0.006). DC-II cells characterized by developed biosynthetic machinery and a large pool of secretory granules storage are increased with the ne in winter. However, LC-I cells showing frequent cytoplasmic degranulation are predominant with glc in summer. Thus, important cellular remodelling occurs in the dromedary PI that may depend upon, or perhaps anticipate, external living conditions.     

Acculturation, childhood trauma and the cortisol awakening response in Mexican-American adults

Exposure to chronic and traumatic stress has been associated with the dysregulation of crucial stress response systems. Acculturation has been associated with unique forms of chronic psychosocial stress. The purpose of this study was to examine the effects of exposure to early traumatic stress and acculturation on dysregulation of the cortisol awakening response (CAR) in Mexican-American adults. Salivary cortisol samples were collected at awakening and 30, 45, and 60 min thereafter, on two consecutive weekdays from 59 healthy Mexican-American adult males (26) and females (33), ages 18-38 years. Participants were assessed for level of acculturation and exposure to early trauma. Data were analyzed using a mixed effects regression model with repeated measures at four time points. Mixed effects regression results indicated a significant Early Trauma × Time interaction (p = .0029) and a significant Acculturation × Time interaction (p = .0015), after controlling for age and sex. Subsequent analyses of the interaction of Trauma × Acculturation × Time showed that more than minimal exposure to either risk factor was associated with attenuation of the awakening cortisol response (p = .0002). Higher levels of acculturation with greater Anglo-orientation were associated with attenuation of the CAR in Mexican-American adults. Both moderate and higher levels of exposure to early trauma were associated with an attenuated CAR. However, greater exposure to both risk factors was only incrementally worse than exposure to either one.     

Synthesis, structure, and magnetic properties of dinuclear nickel(II) complexes with a phenol-based dinucleating ligand with four methoxyethyl chelating arms

Dinuclear nickel(II) complexes [Ni₂(bomp)(MeCO₂)₂]BPh₄ (1) and [Ni₂(bomp)(PhCO₂)₂]BPh₄ (2) were synthesized with the dinucleating ligand 2,6-bis[bis(2-methoxyethyl)aminomethyl]-4-methylphenol [H(bomp)]. X-Ray analysis revealed that the complex 1 · 0.5CHCl₃ contains two nickel(II) ions bridged by phenolic oxygen and two acetate groups, forming a μ-phenoxo-bis(μ-acetato)dinickel(II) core. Electronic spectra were investigated for 1 and 2 in the range of 400-1800 nm, and the data were typical for the octahedral high-spin nickel(II) complexes. Obtained spectral components were well simulated based on the angular overlap model assuming the trigonally distorted octahedral geometry. Magnetic susceptibility was measured for 1 and 2 over a temperature range of 4.5-300 K. The optimized magnetic data were J = 1.75 cm⁻¹, zJ′ = −0.234 cm⁻¹, g = 2.21, D = 15.1 cm⁻¹, and TIP = 370 × 10⁻⁶ cm⁻¹ for complex 1 and J = 3.55 cm⁻¹, zJ′ = −0.238 cm⁻¹, g = 2.23, D = 21.8 cm⁻¹, and TIP = 470 × 10⁻⁶ cm⁻¹ for complex 2. The data revealed ferromagnetic interactions between the two nickel(II) ions.     

Eye Tracking,Cortisol, and a Sleep vs. Wake Consolidation Delay: Combining Methods to Uncover an Interactive Effect of Sleep and Cortisol on Memory

Although rises in cortisol can benefit memory consolidation, as can sleep soon after encoding, there is currently a paucity of literature as to how these two factors may interact to influence consolidation. Here we present a protocol to examine the interactive influence of cortisol and sleep on memory consolidation, by combining three methods: eye tracking, salivary cortisol analysis, and behavioral memory testing across sleep and wake delays. To assess resting cortisol levels, participants gave a saliva sample before viewing negative and neutral objects within scenes. To measure overt attention, participants’ eye gaze was tracked during encoding. To manipulate whether sleep occurred during the consolidation window, participants either encoded scenes in the evening, slept overnight, and took a recognition test the next morning, or encoded scenes in the morning and remained awake during a comparably long retention interval. Additional control groups were tested after a 20 min delay in the morning or evening, to control for time-of-day effects. Together, results showed that there is a direct relation between resting cortisol at encoding and subsequent memory, only following a period of sleep. Through eye tracking, it was further determined that for negative stimuli, this beneficial effect of cortisol on subsequent memory may be due to cortisol strengthening the relation between where participants look during encoding and what they are later able to remember. Overall, results obtained by a combination of these methods uncovered an interactive effect of sleep and cortisol on memory consolidation.     

Effect of environmental enrichment and herbal compound supplementation on physiological stress indicators (chromogranin A,cortisol and tumour necrosis factor-α) in growing pigs

Stress response induces physiological, behavioural, immunological and biochemical changes that directly affect health and well-being. Provision of environmental enrichment and herbal compounds may reduce stress in current commercial pig husbandry systems. The aim of this study was to evaluate the effect of providing different environmental enrichment materials (EE) and a herbal compound (HC) on physiological indicators of acute and chronic stress in growing pigs (salivary cortisol and chromogranin A (CgA), hair cortisol and tumour necrosis factor-α (TNF-α)). Salivary cortisol and CgA have been reported as biomarkers basically of acute stress, whereas hair cortisol and TNF-α have been more related to chronic stress. For this purpose, eight groups of seven pigs each (14 pigs/treatment, 56 pigs in total) were used: (a) two EE groups, (b) two groups supplemented with HC, (c) two groups provided both with EE and HC and (d) two control groups. Samples of hair, saliva and blood were taken to measure cortisol (in hair and saliva), CgA (in saliva) and TNF-α (in blood) at three different times: before starting the experiment (T0), and after 1 (T1) or 2 months (T2) of providing the materials and herbal compound. No differences were found at T0 in salivary or hair cortisol, CgA or TNF-α, whilst at T2, the control group showed significant increased concentrations of CgA and hair cortisol, when compared with the rest of the treatments (P<0.001). These differences were significant at T1 only for CgA (P<0.001). Furthermore, an overall correlation was reported between hair cortisol and salivary CgA (r=0.48, P<0.001). These results support that providing enrichment material or an herbal compound may reduce stress in growing pigs. Furthermore, the results support that hair cortisol and CgA may be proper non-invasive tools to detect stress, specially associated with factors of chronic exposure.     

Urinary free cortisone, but not cortisol, is associated with urine volume in severe obesity

Background

High urine volume enhances urinary free cortisol (UFF) and cortisone (UFE) excretion rates in normal-weight adults and children. Renal excretion rates of glucocorticoids (GC) and their metabolites are frequently altered in obesity. The aim of the present study was to investigate whether UFF and UFE excretion is also affected by urine volume in severely obese subjects.

Experimental

In 24-h urine samples of 59 extremely obese subjects (mean BMI 45.3 ± 8.9 kg/m²) and 20 healthy lean subjects (BMI 22.1 ± 1.8 kg/m²), UFF and UFE, tetrahydrocortisol (THF), 5α-tetrahydrocortisol (5α-THF), and tetrahydrocortisone (THE) were quantified by RIA. The sum of THF, 5α-THF, and THE (GC3), the three major GC metabolites, reflects daily cortisol secretion. 11β-Hydroxysteroid dehydrogenase type 2 (11β-HSD2) activity was assessed by the ratio UFE/UFF. Daily GC excretion rates were corrected for urine creatinine and adjusted for gender and body weight.

Results

In extremely obese subjects, urine volume was significantly associated with creatinine-corrected UFE and 11β-HSD2 activity after adjustment for gender and BMI (r = 0.47, p = 0.0002 and r = 0.31, p = 0.02, respectively). However, urine volume was not associated with creatinine-corrected UFF and GC3 (p = 0.4 and p = 0.6, respectively). In lean controls, urine volume was significantly associated with creatinine-corrected UFE and UFF (r = 0.58, p = 0.01 and r = 0.55, p = 0.02, respectively), whereas urine volume was not associated with 11β-HSD2 activity after appropriate adjustment (p = 0.3).

Conclusions

In severe obesity, in contrast to normal weight, renal excretion of UFE, but not of UFF is affected by fluid intake. This discrepancy may be due to the increased renal 11β-HSD2 activity in obesity.     

An in vivo microdialysis coupled with liquid chromatography/tandem mass spectrometry study of cortisol metabolism in monkey adipose tissue

It is postulated that elevated tissue concentrations of cortisol may be associated with the development of metabolic syndrome, obesity, and type 2 diabetes. The 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) enzyme regenerates cortisol from inactive cortisone in tissues such as liver and adipose. To better understand the pivotal role of 11β-HSD1 in disease development, an in vivo microdialysis assay coupled with liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis using stable isotope-labeled (SIL) cortisone as a substrate was developed. This assay overcomes the limitations of existing methodologies that suffer from radioactivity exposure and analytical assay sensitivity and specificity concerns. Analyte extraction efficiencies (E_d) were evaluated by retrodialysis. The conversion of SIL-cortisone to SIL-cortisol in rhesus monkey adipose tissue was studied. Solutions containing 100, 500, and 1000 ng/mL SIL-cortisone were locally delivered through an implanted 30-mm microdialysis probe in adipose tissue. At the delivery rate of 1.0 and 0.5 μL/min, E_d values for SIL-cortisone were between 58.7 ± 5.6% (n = 4) and 72.7 ± 1.3% (n = 4), whereas at 0.3 μL/min E_d reached nearly 100%. The presence of 11β-HSD1 activities in adipose tissue was demonstrated by production of SIL-cortisol during SIL-cortisone infusion. This methodology could be applied to cortisol metabolism studies in tissues of other mammalian species.     

Interruption of the canine estrous cycle with a low and a high dose of the GnRH antagonist, acyline

To test the efficacy and clinical safety of a low and high dose of the GnRH antagonist, acyline, on estrous cycle interruption and anovulation in female dogs, 20 proestrous (<3 d) bitches were randomly assigned to one of the following pharmacological protocols (given sc): acyline 110 μg/kg (ACY-L; n = 6); acyline 330 μg/kg (ACY-H; n = 8); or placebo (PLACE, n = 6). The animals were monitored (clinical and vaginal cytology examinations) daily for 60 d. Blood samples for serum progesterone serum concentrations were collected 14 d after treatment to determine if ovulation had occurred. Appearance of side effects and days to the onset of the first spontaneous estrous cycle after treatment were also recorded. In both ACY groups, but not the PLACE group, estrous cycles were interrupted after treatment (P < 0.05). The interval from treatment to estrus interruption in ACY-L and ACY-H groups was 3.0 ± 0.6 and 3.2 ± 0.2 d, respectively (LSM ± SEM; P > 0.05). In the PLACE bitches, physical, behavioral and cytological proestrus slowly progressed to estrus and diestrus. Ovulation was absent in all ACY, but not in PLACE bitches (P < 0.05). None of the females manifested side effects related to the treatments (P > 0.05). Spontaneous return to a normal estrous cycle during the study period occurred in all ACY (ACY-L 19.5 ± 2.7 d vs ACY-H 24.8 ± 2.0 d; P > 0.05), but in none of the PLACE bitches (P < 0.05). In conclusion, acyline efficiently, safely and reversibly interrupted an early phase of the estrous cycle in bitches by preventing ovulation.