Measuring luteinising hormone pulsatility with a robotic aptamer-enabled electrochemical reader

Assessment of luteinising hormone pulsatility is important in the diagnosis of reproductive disorders. Here the authors develop a DNA aptamer-based electrochemical analysis integrated into a robotic platform for high-throughput and sensitive analysis

The effects of kisspeptin on β-cell function, serum metabolites and appetite in humans

Aims: To investigate the effect of kisspeptin on glucose-stimulated insulin secretion and appetite in humans. Materials and methods: In 15 healthy men (age: 25.2 ± 1.1 years; BMI: 22.3 ± 0.5 kg m−2), we compared the effects of 1 nmol kg−1 h−1 kisspeptin versus vehicle administration on glucose-stimulated insulin secretion, metabolites, gut hormones, appetite and food intake. In addition, we assessed the effect of kisspeptin on glucose-stimulated insulin secretion in vitro in human pancreatic islets and a human β-cell line (EndoC-βH1 cells). Results: Kisspeptin administration to healthy men enhanced insulin secretion following an intravenous glucose load, and modulated serum metabolites. In keeping with this, kisspeptin increased glucose-stimulated insulin secretion from human islets and a human pancreatic cell line in vitro. In addition, kisspeptin administration did not alter gut hormones, appetite or food intake in healthy men. Conclusions: Collectively, these data demonstrate for the first time a beneficial role for kisspeptin in insulin secretion in humans in vivo. This has important implications for our understanding of the links between reproduction and metabolism in humans, as well as for the ongoing translational development of kisspeptin-based therapies for reproductive and potentially metabolic conditions

Supplementary figures.

Fig A: Testing HormoneBayes on synthetic data. Fig B: Assessing the effect of the prior for the LH clearance rate. Fig C: Tuning HormoneBayes when pulses are not clear by using a more informative prior on parameter f. Fig D: Pulse identification using HormoneBayes. Fig E: Using HormoneBayes to identify the effect of interventions on LH pulsatility. (PDF)</p

HormoneBayes handles LH pulsatility analysis in different contexts.

(A) Inferred pulsatility strength and maximum secretion rate parameters for different individuals: healthy men (n = 10); healthy post-menopause women (n = 13); healthy pre-menopausal women (n = 4). (B) Inferred parameters for healthy pre-menopausal women (n = 4); women with PCOS (n = 6) and women with HA (n = 5) illustrating how the assessment of LH pulsatility could help facilitate diagnosis of patients presenting with reproductive endocrine disorders. (C) Representative fits of the model are given for one subject in each dataset.</p

Pulse identification using HormoneBayes.

(A) Pulses can be identified using the expected value of the pulsatile hypothalamic signal, which can be interpreted as the probability of a pulse at a given timepoint. Here, we mark the onset of a pulse when the pulsatile hypothalamic signal crosses the 0.5 threshold, indicating that at this point a pulse is the most likely event. (B) The majority of the identified pulses (89%, 77/87) are in line with those obtained using the deconvolution method. For the analysis we used LH data obtained from healthy pre-menopausal women in early follicular phase (n = 16).</p

pcbi.1011928.t001 - HormoneBayes: A novel Bayesian framework for the analysis of pulsatile hormone dynamics

pcbi.1011928.t001 - HormoneBayes: A novel Bayesian framework for the analysis of pulsatile hormone dynamics</p

HormoneBayes: a Bayesian framework for analyzing pulsatile LH dynamics.

The framework uses a parsimonious mathematical model to describe LH levels in circulation as the net effect of secretion and clearance. In the model secretion is driven by a basal hypothalamic signal and a pulsatile signal (mimicking the dynamics of the GnRH pulse generator which can be turned ‘on’ or ‘off’). An efficient Markov-Chain Monte-Carlo (MCMC) method performs the Bayesian inference and extracts model parameters and latent hypothalamic dynamics, which are compatible with the observed data.</p

Starship:Study of Telomeres and Role of Sex Hormones in Pulmonary Fibrosis

RATIONALE Pulmonary fibrosis is an interstitial lung disease (ILD) feature with no curative (other than lung transplantation) or preventative treatments. Recent evidence supports a causal role of short telomeres, and our pre-print findings suggest a potentially protective interaction between optimised sex hormone concentration (testosterone and oestrogen) in men and women and telomere maintenance pathways1. Sex hormone bioavailability is regulated by sex hormone binding globulin (SHBG) in both sexes. Supra-physiological dosing of testosterone in men causes erythrocytosis, so monitoring of haemoglobin concentration and haematocrit is required. STARSHIP set out to assess whether future therapeutic trials of sex hormone supplementation in ILD could be safe and warranted without causing supra-physiological dosing. METHODS Venepuncture was performed on 102 outpatients (age 49-89y, male n=80 [78%]) with fibrotic ILD a ta single regional centre in the UK. Age and sex-matched control samples were obtained from the Exeter 10,000 (EXTEND) biobank. Blood leukocyte telomere length (LTL; measured using high-throughput single telomere length analysis) and sex hormone/SHBG concentrations were determined in all samples. Patients consented for access to their clinical data (including routine pulmonary function). In-depth interviews were carried out by telephone with 93 self-selecting patients, using structured interviews with questions informed by clinical practice, the research team ,and patient partners. RESULTS Almost all male ILD patients (n=77/79) had haemoglobin concentrations and haematocrit below the upper reference limit. Compared to age-matched controls (AMCs) mean LTL was shorter for patients (4.57kb [95%CI: 4.46-4.69] vs 4.78kb [95%CI:4.67-4.89], p&lt;0.006). Mean bioavailable testosterone was lower in male patients (4.95nmol/L[95%CI: 4.50-5.41] vs 6.40nmol/L [95%CI: 5.82-6.98] in AMC; p&lt;0.0001, n=80). Oestrogen concentrations were low for both female patients and controls in keeping with their post-menopausal age, with higher SHBG concentrations for patients (mean 73.7nmol/L [95%CI: 58.6-88.9] vs 52.9nmol/L [95%CI: 42.0-63.8] in AMC; p=0.022). Mean free androgen index (FAI) was below the reference range for female patients (mean 0.51 [95%CI: 0.35-0.67] vs 1.23 [95%CI: 0.77-1.69] in AMC; p= 0.0036, N=22). Both DLCO% predicted (p=0.0005) and FVC% predicted(p=0.005) correlated with bioavailable testosterone concentration in male patients (age-adjusted analysis). FVC% predicted associated with FAI in females (p=0.013) but no association was seen between FAI and DLCO% predicted. CONCLUSIONS Interventional trials of sex hormone supplementation without causing supra-physiological dosing warrant further exploration in patients with ILD

Starship:Study of Telomeres and Role of Sex Hormones in Pulmonary Fibrosis

RATIONALE Pulmonary fibrosis is an interstitial lung disease (ILD) feature with no curative (other than lung transplantation) or preventative treatments. Recent evidence supports a causal role of short telomeres, and our pre-print findings suggest a potentially protective interaction between optimised sex hormone concentration (testosterone and oestrogen) in men and women and telomere maintenance pathways1. Sex hormone bioavailability is regulated by sex hormone binding globulin (SHBG) in both sexes. Supra-physiological dosing of testosterone in men causes erythrocytosis, so monitoring of haemoglobin concentration and haematocrit is required. STARSHIP set out to assess whether future therapeutic trials of sex hormone supplementation in ILD could be safe and warranted without causing supra-physiological dosing. METHODS Venepuncture was performed on 102 outpatients (age 49-89y, male n=80 [78%]) with fibrotic ILD a ta single regional centre in the UK. Age and sex-matched control samples were obtained from the Exeter 10,000 (EXTEND) biobank. Blood leukocyte telomere length (LTL; measured using high-throughput single telomere length analysis) and sex hormone/SHBG concentrations were determined in all samples. Patients consented for access to their clinical data (including routine pulmonary function). In-depth interviews were carried out by telephone with 93 self-selecting patients, using structured interviews with questions informed by clinical practice, the research team ,and patient partners. RESULTS Almost all male ILD patients (n=77/79) had haemoglobin concentrations and haematocrit below the upper reference limit. Compared to age-matched controls (AMCs) mean LTL was shorter for patients (4.57kb [95%CI: 4.46-4.69] vs 4.78kb [95%CI:4.67-4.89], p&lt;0.006). Mean bioavailable testosterone was lower in male patients (4.95nmol/L[95%CI: 4.50-5.41] vs 6.40nmol/L [95%CI: 5.82-6.98] in AMC; p&lt;0.0001, n=80). Oestrogen concentrations were low for both female patients and controls in keeping with their post-menopausal age, with higher SHBG concentrations for patients (mean 73.7nmol/L [95%CI: 58.6-88.9] vs 52.9nmol/L [95%CI: 42.0-63.8] in AMC; p=0.022). Mean free androgen index (FAI) was below the reference range for female patients (mean 0.51 [95%CI: 0.35-0.67] vs 1.23 [95%CI: 0.77-1.69] in AMC; p= 0.0036, N=22). Both DLCO% predicted (p=0.0005) and FVC% predicted(p=0.005) correlated with bioavailable testosterone concentration in male patients (age-adjusted analysis). FVC% predicted associated with FAI in females (p=0.013) but no association was seen between FAI and DLCO% predicted. CONCLUSIONS Interventional trials of sex hormone supplementation without causing supra-physiological dosing warrant further exploration in patients with ILD