Long-Term Combined Effects of Citrulline and Nitrate-Rich Beetroot Extract Supplementation on Recovery Status in Trained Male Triathletes: A Randomized, Double-Blind, Placebo-Controlled Trial

Citrulline (CIT) and nitrate-rich beetroot extract (BR) are widely studied ergogenic aids. Nevertheless, both supplements have been studied in short-term trials and separately. To the best of the authors’ knowledge, the effects of combining CIT and BR supplementation on recovery status observed by distance covered in the Cooper test, exercise-induced muscle damage (EIMD) and anabolic/catabolic hormone status have not been investigated to date. Therefore, the main purpose of this research was to assess the effect of the long-term (9 weeks) mixture of 3 g/day of CIT plus 2.1 g/day of BR (300 mg/day of nitrates (NO3−)) supplementation on recovery by distance covered in the Cooper test, EIMD markers (urea, creatinine, AST, ALT, GGT, LDH and CK) and anabolic/catabolic hormones (testosterone, cortisol and testosterone/cortisol ratio (T/C)) in male trained triathletes. Thirty-two triathletes were randomized into four different groups of eight triathletes in this double-blind, placebo-controlled trial: placebo group (PLG), CIT group (CITG; 3 g/day of CIT), BR group (BRG; 2.1 g/day of BR (300 mg/day of NO3−)) and CIT-BR group (CIT-BRG; 3 g/day of CIT plus 2.1 g/day of BR (300 mg/day of NO3−)). Distance covered in the Cooper test and blood samples were collected from all participants at baseline (T1) and after 9 weeks of supplementation (T2). There were no significant differences in the interaction between group and time in EIMD markers (urea, creatinine, AST, ALT, GGT, LDH and CK) (p > 0.05). However, significant differences were observed in the group-by-time interaction in distance covered in the Cooper test (p = 0.002; η2p = 0.418), cortisol (p = 0.044; η2p = 0.247) and T/C (p = 0.005; η²p = 0.359). Concretely, significant differences were observed in distance covered in the Cooper test percentage of change (p = 0.002; η²p = 0.418) between CIT-BRG and PLG and CITG, in cortisol percentage change (p = 0.049; η2p = 0.257) and in T/C percentage change (p = 0.018; η2p = 0.297) between CIT-BRG and PLG. In conclusion, the combination of 3 g/day of CIT plus 2.1 g/day of BR (300 mg/day of NO3−) supplementation for 9 weeks did not present any benefit for EIMD. However, CIT + BR improved recovery status by preventing an increase in cortisol and showing an increase in distance covered in the Cooper test and T/C.

Novel Opipramol-Baclofen Combination Alleviates Depression and Craving and Facilitates Recovery From Substance Use Disorder—An Animal Model and a Human Study

Substance use disorders (SUDs) are associated with depression and anxiety, with the latter being one of the major factors in substance-seeking and relapse. Due to dose-dependent sedative side effects there is limited efficacy of baclofen treatment for SUDs. Here we suggest the use of a novel combination of opipramol and baclofen (O/B) which is known to attenuate anxiety and depression, for the facilitation of recovery from SUDs. Since both opipramol and baclofen have a common downstream signal transduction, their individual doses could be reduced while still maintaining the benefits of the combination. We tested the O/B combination in both animals and patients. Rats treated with O/B showed significant attenuation in craving behavior and in relapse rate during withdrawal from cocaine. In a double-blind, placebo-controlled pilot study, conducted in a residential detoxification center, 14 males and 3 females, aged 28–60 years were assigned to a study (n = 6) and a placebo (n = 11) group (placebo group: 40 ± 10.5 years; O/B group 40 ± 10.8 years). The participants completed scales measuring depression, anxiety and craving symptoms and provided saliva samples for stress hormone examination [cortisol and dehydroepiandrosterone-sulfate (DHEA-S)]. Participants with polysubstance use disorder (PsUD) treated with O/B showed a reduction in cravings and depression and an increase in DHEA-S and in the DHEA-S/cortisol ratio. Our findings indicate a beneficial effect of O/B treatment. This study suggests a novel candidate for pharmacological treatment of patients with SUD and comorbid mood/anxiety disorders that may facilitate their rehabilitation.

Effects of Wash Protocol and Contamination Level on Concentrations of Cortisol and Dehydroepiandrosterone (DHEA) in Swine Hair

The effect of washing procedure and contamination level on the concentrations of cortisol and dehydroepiandrosterone (DHEA) in swine hair was explored over two studies. Hair shaved from finisher pigs (n = 8) and sows (n = 8, cortisol study 1 only) was split into two treatments (two hair samples/pig) to receive either three isopropanol or methanol washes, and two paired subsamples of hair were contaminated with feces and urine, mildly or severely. Samples were further subdivided and received one, three, or five methanol washes. Hormone concentrations were quantified from the hair and wash solvent, and the ratio of hormones in the solvent to that in the hair calculated. When grouping sow and grower hair together for analysis, hair cortisol concentrations were 13% greater after three isopropanol washes compared to methanol (22.84 ± 3.12 vs. 19.77 ± 2.64 pg/mg, respectively). When analyzing sow and grower hair separately, sow hair cortisol concentrations were 20% higher following three isopropanol washes compared to methanol washes (22.06 ± 5.21 vs. 27.72 ± 5.65 pg/mg), with no differences in grower pig hair concentrations. The solvent cortisol concentrations did not differ with wash solvent. No differences were seen for DHEA. Contamination level did not influence hormone concentrations. Hair cortisol concentrations were 24% higher after one wash compared to five washes (11.98 ± 1.47 vs. 9.05 ± 0.92 pg/mg,), whereas the solvent cortisol concentrations were 80% and 84% higher after one wash compared to three and five washes, respectively (21.09 ± 4.04 vs. 4.21 ± 1.62 vs. 3.36 ± 1.32 pg/mg). The solvent–hair cortisol ratio was 65% and 73% higher following one wash compared to three and five washes (1.36 ± 0.80 vs. 0.47 ± 0.12 vs. 0.37 ± 0.14). Hair DHEA concentrations were 39% higher after one wash compared to five washes (42.39 ± 6.87 vs. 26.02 ± 5.69 pg/mg). The solvent DHEA concentrations, and the solvent–hair ratio for DHEA were 94% and 98% and 92% and 98% higher going from one wash to three and five washes, respectively (solvent: 5.07 ± 0.26 vs. 0.28 ± 0.12 vs. 0.12 ± 0.09 pg/mg and solvent–hair ratio: 0.13 ± 0.006 vs. 0.010 ± 0.004 vs. 0.003 ± 0.002). Following three methanol washes, the non-contaminated hair had 46% and 48% higher hair (17.47 ± 1.12 vs. 9.35 ± 0.80 vs. 9.05 ± 1.06 pg/mg) and a 76% and 72% higher solvent (16.31 ± 8.07 vs. 3.92 ± 0.50 vs. 4.50 ± 2.31 pg/mg) cortisol concentration compared to mild and severely contaminated hair, respectively. Wash solvent influences cortisol concentrations in swine hair, but not DHEA. Contaminated swine hair should be avoided in analyses when possible.

Impact of Optimal Timing of Intake of Multi-Ingredient Performance Supplements on Sports Performance, Muscular Damage, and Hormonal Behavior across a Ten-Week Training Camp in Elite Cyclists: A Randomized Clinical Trial

Multi-ingredient performance supplements (MIPS), ingested pre- or post-workout, have been shown to increase physiological level effects and integrated metabolic response on exercise. The purpose of this study was to determine the efficacy of pre-and post-training supplementation with its own MIPS, associated with CHO (1 g·kg−1) plus protein (0.3 g·kg−1) on exercise-related benchmarks across a training camp for elite cyclists. Thirty elite male cyclists participated in a randomized non-placebo-controlled trial for ten weeks assigned to one of three groups (n = 10 each): a control group treated with CHO plus protein after training (CG); a group treated with MIPS before training and a CHO plus protein after training, (PRE-MIPS); a group treated with CHO plus protein plus MIPS after training, (POST-MIPS). Performance parameters included (VO2max, peak; median and minimum power (W) and fatigue index (%)); hormonal response (Cortisol; Testosterone; and Testosterone/Cortisol ratio); and muscle biomarkers (Creatine kinase (CK), Lactate dehydrogenase (LDH), and Myoglobin (Mb)) were assessed. MIPS administered before or after training (p ≤ 0.05) was significantly influential in attenuating CK, LDH, and MB; stimulating T response and modulating C; and improved on all markers of exercise performance. These responses were greater when MIPS was administered post-workout.

First Trimester Maternal Cortisol Signatures in Small-for-gestational Age

Background: Abnormal maternal hypothalamus-pituitary-adrenal axis is associated with fetal growth, and we hypothesized that the alteration in metabolic signatures of cortisol might be detectable during early pregnancy. The objective of this study was to identify predictable maternal serum signatures of cortisol metabolism during the first trimester of women who are expected to deliver small-for-gestational age (SGA) neonates.Methods: This prospective cohort study included 112 pregnant women (with and without SGA, n = 56 each). Maternal serum samples were collected at 10~14 gestational weeks to quantify the levels of cortisol and its precursors and metabolites by liquid chromatography-mass spectrometry.Results: Increased maternal serum levels of tetrahydrocortisol (THF, 11.82 ± 8.16 ng/mL vs. 7.51 ± 2.90 ng/mL, P < 0.005) and decreased 21-deoxycortisol (21-deoxyF, 2.98 ± 1.36 ng/mL vs. 4.33 ± 2.06 ng/mL, P < 0.0001) were observed in pregnant women carrying SGA fetus. In conjunction with individual steroid levels, metabolic ratios corresponding to the activity of related enzymes were calculated. In addition to increased THF/cortisol ratio (P < 0.006), the SGA group showed a significant increase in the two metabolic ratios including cortisol/11-deoxycortisol (F/11-deoxyF; P < 0.03) and cortisol/21-deoxycortisol (F/21-deoxyF; P < 0.0003) indicating cortisol biosynthesis. The ROC curve generated in combination with three variables of 21-deoxyF concentration and two metabolic ratios of F/21-deoxyF and THF/F resulted in AUC = 0.824 (95% confidence interval, 0.713 ~ 0.918). Conclusions: A significant decrease in maternal serum levels of 21-deoxyF and an increase in two metabolic ratios of F/21-deoxyF and THF/F, indicating cortisol biosynthetic rate, represent a reliable biomarker for the prediction of SGA in the first trimester.

EVALUATION OF THE BIOLOGICAL RELATIONSHIPS OF CORTICOSTERONE AND CORTISOL IN THE BODY OF CHICKENS DURING A STRESS REACTION

The variability of corticosterone and cortisol concentrations and their ratio in laying hens during the development of a stress reaction, as well as the correlation of hormone levels with the variability of protein parameters in the blood, are evaluated. It was found that when the technological factor (the density of birds in the cage) changes by 1.5 times, the amount of glucocorticoids in the blood of birds during the stress reaction increases, both due to corticosterone and cortisol. Therefore, the balance between the hormones were not significantly changed and is in the range of 4.23-5,43conv. Units. When the technological factor is increased by 2 times, the content of corticosterone in the blood of birds mainly increases, determining an increase in the value of the corticosterone/cortisol ratio by 16.56-28.71%. The biological effects of glucocorticoids in the body of birds are realized by corticosterone, the level of which in a moderate and strong degree, including statistically significant, both under normal and stress conditions, is associated with the activity of transamination enzymes: Corticosterone-AlAT (r(group I) = 0,69±0,32 – 0,88±0,21; r(group II) =0.63±0.24 – 0.91±0.13; r(group III) = 0,59±0,22 – 0,76±0,16) and Corticosterone-AsAT (r(I group) = 0,57±0,36 – 0,83±0,25; r(group II) =0,68±0,23 – 0,72±0,22; r(group III) = 0,67±0,19 – 0,89±0,12), and also with the concentration of albumins during the development of a stress reaction (Corticosterone-Alb: r(group II) = -0,69±0,22 – -0,81±0,19; r(group III) = -0,58±0,21 – -0,81±0,16).

Epinephrine to the Rescue: Identifying the Offending Side in Bilateral (B/L) Macronodular Adrenal Hyperplasia (BMAH) With Cushing Syndrome (CS) and Hyperaldosteronism

Management of ACTH-independent CS in the setting of B/L adrenal masses can be challenging. We present a patient with concomitant cortisol and aldosterone hypersecretion and B/L adrenal masses who was treated with unilateral adrenalectomy (UA) based on simultaneous B/L adrenal vein sampling (AVS) results normalized to plasma epinephrine levels, to correct for flow and dilution. A 60-year-old lady presented for evaluation of incidentally discovered B/L adrenal masses. CT abdomen with contrast revealed B/L lobulated adrenal glands measuring 3.4 x 5.5 x 4.1 cm on the right and 3.8 x 4.2 x 2 cm on the left (pre contrast density of 37 and 44 Hounsfield Units respectively). She had a recent diagnosis of HTN and prediabetes. She reported fatigue, easy bruising, and muscle weakness. Family history was notable for a sister with CS and B/L adrenal masses who was successfully treated with UA. On physical examination her BP was 160/94 mm Hg and she did not have an obvious cushingoid appearance. Laboratory testing included an elevated AM serum cortisol level of 16.6 mcg/dL following 1 mg of dexamethasone given at 11 pm the previous night. ACTH was &lt;1.0 pg/mL consistent with ACTH-independent CS. She had a serum aldosterone level of 26.6 ng/dL(4.0 - 31.0 ng/dL),simultaneous plasma renin activity of 0.5 ng/ml/hr (0.5–4 ng/ml/hr) and serum potassium level of 4.4 mmol/l. She underwent B/L simultaneous AVS. Given concomitant hyperaldosteronism, plasma epinephrine levels were measured to determine successful catheter placement and to correct for flow and dilution between the adrenal veins. Right adrenal vein to peripheral vein cortisol ratio was 9.27 (&gt; 6.5) consistent with cortisol hypersecretion. The right to left cortisol ratio was 2.3 (&gt;/= 2.3), consistent with right sided predominance. Right to left ratio of the epinephrine corrected cortisol level was 2.7. She underwent right adrenalectomy, and pathology showed macronodular adrenal hyperplasia. Postoperatively, she had resolution of HTN and normalization of 24 hour urine free cortisol. UA is increasingly becoming recognized as a successful treatment for BMAH due to lower risk of adrenal crisis when compared to B/L adrenalectomy. In a recent publication, remission from CS was seen in 94.4 % of patients with BMAH following UA, with recurrence in 19% of patients. The mean time to recurrence was 5 years. We utilized cutoff values that were previously reported for cortisol lateralization in interpreting AVS. Our patient attained remission of CS 9 months post UA. Epinephrine levels were not significantly different between the adrenal veins, possibly due to simultaneous AVS, so we were also able to calculate epinephrine corrected cortisol ratios. Simultaneous B/L AVS with epinephrine to localize autonomous hypercortisolism in the setting of concomitant hyperaldosteronism can be beneficial in guiding surgical management of patients.

Exercise stress assessed by automated measurements of salivary testosterone and cortisol levels and testosterone/cortisol ratio within circadian rhythms in male long-distance runners

Background: Exercise stress promoted cortisol and testosterone secretions that have their own circadian rhythms. It is necessary that the testosterone, cortisol and testosterone/cortisol ratio (T/C ratio) are measured through their rhythms for use in monitoring exercise-induced stress in athletes. Automated measurement has not been applied to salivary testosterone, which requires passive drooling difficult to collect sufficient saliva rapidly. This study aimed to verify whether automated measurements of the testosterone and cortisol concentrations and T/C ratio using saliva collected sequentially can effectively assess exercise intensity differences within circadian rhythms in male athletes.Methods: We investigated the correlations of testosterone and cortisol concentrations measured by electrochemiluminescence immunoassay (ECLIA) between saliva and sera collected from 20 male long-distance runners. We collected the runners' saliva sequentially by passive drooling on two consecutive days involving different intensity trainings in the morning and evening; salivary testosterone and cortisol concentrations were measured by ECLIA. Each exercise intensity was measured by running distances, velocities, Borg scale score and maximum pulse rate during exercise.Results: The salivary testosterone and cortisol concentrations were positively correlated with the respective total serum hormone concentrations. The runners were divided into low-intensity exercise group (n = 8) and high-intensity exercise group (n = 7), in which five runners were excluded because measurable saliva samples could not be obtained due to low volume and high-viscosity. Sequential saliva collection and automated measurements detected the runners' circadian rhythms of testosterone, cortisol and T/C ratio. The rate of change in the salivary cortisol concentrations were significantly higher and that in the T/C ratio was significantly lower in the evening interval training on day 1 in the high-intensity exercise group which had significantly higher running velocity, Borg scale score, and maximum pulse rate values; this relationship was not shown for salivary testosterone.Conclusions: Automated measurements of the salivary cortisol concentration and the T/C ratio reflected different exercise intensities may be useful for creating appropriate exercise programs for athletes. Conversely, the automated measurements of salivary testosterone and T/C ratio, which require passive drooling to collect saliva, may be less suitable for practical use with athletes than the salivary cortisol-only measurement