Yasin Y. Dhaher

A bstract: Dynamic endocrine transitions govern female physiology, yet several reproductive hormones remain incompletely characterized across the natural menstrual cycle. Relaxin, a peptide hormone classically associated with pregnancy, has recognized vascular, extracellular matrix, and anti-fibrotic actions, but its physiology in nonpregnant women remains poorly defined. We performed dense longitudinal endocrine profiling across 80 natural menstrual cycles from 34 healthy women, complemented by oral contraceptive users, to define circulating relaxin-2 dynamics relative to ovarian steroids. Using generalized additive modeling of cycle-normalized trajectories, we identified relaxin as a highly phase-locked late-luteal hormone with a sharp, reproducible peak distinct from estradiol and progesterone. Progesterone rose earlier after ovulation and peaked at cycle day 23.9, whereas relaxin peaked significantly later at day 28.0, revealing an approximately 4-day temporal offset. Relaxin showed lower inter-cycle variability and narrower temporal dispersion than progesterone, indicating unexpectedly precise endocrine timing. Detectable relaxin persisted into early menses, suggesting continued luteal peptide release across the menstrual boundary. In combined oral contraceptive users, endogenous cyclicity was attenuated and circulating relaxin was effectively absent despite exogenous progestin exposure, implicating ovulation-dependent corpus luteum biology rather than steroid exposure alone as the primary determinant of relaxin secretion. These findings identify relaxin as a reproducible component of normal cyclic endocrine physiology rather than a hormone restricted to pregnancy. The discrete late-luteal relaxin window establishes a previously under-resolved temporal feature of the menstrual cycle and provides a framework for investigating its reproductive, vascular, and connective-tissue significance.

Tracking individual motor units (MUs) across multiple testing visits is critical for understanding neuromuscular adaptations and disease progression. However, existing methods fail to account reliably for spatiotemporal variations in motor unit action potentials (MUAPs) and lack validation for longitudinal tracking. Here, we present a robust computational framework for continuous MU tracking that integrates automatic estimation of electrode grid displacements and compensation for MUAP shape variations. Our algorithm demonstrated superior tracking accuracy, capturing 63% and 75% of MUs in the tibialis anterior and medial gastrocnemius, respectively, substantially outperforming state-of-the-art methods. By applying this approach to investigate estradiol-driven neuromuscular plasticity as a model system, we reveal previously undetectable modulations in recruitment thresholds and firing rates, linking hormonal fluctuations to motor unit behavior. Beyond advancing fundamental neuromuscular research, our approach holds promise for enhancing biomarker sensitivity in clinical trials, particularly in conditions such as amyotrophic lateral sclerosis, where reliable MU tracking is paramount. These findings establish a new standard for non-invasive motor unit tracking, unlocking opportunities for longitudinal neurophysiological assessment and evaluation of therapeutics.

Objective. Non-invasive spinal stimulation has the potential to modulate spinal excitability. This study explored the modulatory capacity of sub-motor grid-based transcutaneous spinal cord stimulation (tSCS) applied to the lumbar spinal cord in neurologically intact participants. Our objective was to examine the effect of grid spinal stimulation on polysynaptic reflex pathways involving motoneurons and interneurons likely activated by Aβ/δ fiber-mediated cutaneous afferents. Approach. Stimulation was delivered using two grid electrode montages, generating a net electric field in transverse or diagonal directions. We administered tSCS with the center of the grid aligned with the T10–T11 spinous process. Participants were seated for the 20 min stimulation duration. At 30 min after the cessation of spinal stimulation, we examined neuromodulatory effects on spinal circuit excitability in the tibialis anterior muscle by employing the classical flexion reflex paradigms. Additionally, we evaluated spinal motoneuron excitability using the H-reflex paradigm in the soleus muscle to explore the differential effects of tSCS on the polysynaptic versus monosynaptic reflex pathway and to test the spatial extent of the grid stimulation. Main results. Our findings indicated significant neuromodulatory effects on the flexion reflex, resulting in a net inhibitory effect, regardless of the grid electrode montages. Our data further indicated that the flexion reflex duration was significantly shortened only by the diagonal montage. Significance. Our results suggest that grid-based tSCS may specifically modulate spinal activities associated with polysynaptic flexion reflex pathways, with the potential for grid-specific targeted neuromodulation.

The onset of degenerative joint diseases such as post-traumatic osteoarthritis (PTOA) are associated with joint injury, biomechanical changes, and synovial biochemical anomalies. Sex and reproductive endocrinology have been emerging as potential risk factors, with epidemiological evidence revealing that female’s exhibit higher PTOA risk and poorer outcomes post-injury compared to males. Sex hormones, including estradiol, progesterone, and testosterone, have been shown to regulate inflammatory signaling in immune and synovial cells, yet their collective impact on injury-induced joint inflammation and catabolism is poorly understood. Using an in silico kinetic model, we investigated the effects of sex-specific endocrine states on post-injury mechanisms in the human synovial joint. Our model results reveal that heightened estradiol levels in pre-menopausal females during the peri-ovulatory phase increase interleukin (IL)-1β expression and suppress IL-10 expression within the synovium after a simulated injury. Conversely, elevated testosterone levels in males decrease post-injury IL-1β, tumor necrosis factor alpha (TNF)-α, and stromelysin (MMP)-3 expression while increasing IL-10 production compared to females. Gaining insight into the effects of sex hormones on injury-induced inflammation and cartilage degradation provides a basis for designing future experimental and clinical studies to explore their effects on the synovial system, with a particular focus on the female sex.

The results from this study have wide-ranging implications for research and treatment in women’s health including considerations in research design and interpretation for studies including women taking oral contraceptives, the potential for more precise and personalized methods of dosing to reduce unwanted side effects and adverse events, and the potential treatment of a variety of disorders ranging from musculoskeletal to neurological with exogenous hormones.

Purpose: The purpose of this study is to characterize the effect of sex and the influence of oral contraception usage on musculotendinous injury (MTI). Current literature suggests a disparity in the incidence of MTI between males and females. This may be attributed to inherent biological differences between the sexes, such as in the sex hormonal milieu. There is a lack of information associating sex hormone milieu and MTI. Methods: We searched the PearlDiver database (a for-fee healthcare database) for males, females taking oral contraceptives (OC), and eumenorrheic females not taking any form of hormonal contraceptives (non-OC) 18–39 yr old. The three populations were matched by age and body mass index. We queried the database for lower-extremity skeletal MTI diagnoses in these groups. Results: Each group contained 42,267 patients with orthopedic injuries. There were a total of 1476 (3.49%) skeletal MTI in the male group, 1078 (2.55%) in non-OC females, and 231 (0.55%) in OC females. Both the non-OC and the OC groups had a significantly smaller proportion of MTI than males (P < 0.0001), and therefore these groups were less likely (adjusted odds ratios, 0.72 and 0.15, respectively) to experience MTI when controlled for potential covariates. Conclusions: In this study, we show that females are less likely to develop MTI to total injuries, when compared with males, with OC using females being least likely followed by non-OC females. These results are consistent with other epidemiological studies; however, overall results in the literature are variable. This study adds to the emerging body of literature on sex hormone–influenced musculoskeletal injury but, more specifically, MTI, which have not been rigorously investigated.

The natural variation in estrogen secretion throughout the female menstrual cycle impacts various organs, including estrogen receptor (ER)-expressed skeletal muscle. Many women commonly experience increased fatigue or reduced energy levels in the days leading up to and during menstruation, when blood estrogen levels decline. Yet, it remains unclear whether endogenous 17β-estradiol, a major estrogen component, directly affects the energy metabolism in skeletal muscle due to the intricate and fluctuating nature of female hormones. In this study, we employed 2D ³¹P FID-MRSI at 7T to investigate phosphoryl metabolites in the soleus muscle of a cohort of young females (average age: 28 ± 6 years, n = 7) during the early follicular (EF) and peri-ovulation (PO) phases, when their blood 17β-estradiol levels differ significantly (EF: 28 ± 18 pg/mL vs. PO: 71 ± 30 pg/mL, p < 0.05), while the levels of other potentially interfering hormones remain relatively invariant. Our findings reveal a reduction in ATP-referenced phosphocreatine (PCr) levels in the EF phase compared to the PO phase for all participants (5.4 ± 4.3%). Furthermore, we observe a linear correlation between muscle PCr levels and blood 17β-estradiol concentrations (r = 0.64, p = 0.014). Conversely, inorganic phosphate Pi and phospholipid metabolite GPC levels remain independent of 17β-estradiol but display a high correlation between the EF and PO phases (p = 0.015 for Pi and p = 0.0008 for GPC). The robust association we have identified between ATP-referenced PCr and 17β-estradiol suggests that 17β-estradiol plays a modulatory role in the energy metabolism of skeletal muscle.

It has been shown that estrogen and progesterone receptors are expressed in the spinal cord; therefore, fluctuation in their concentrations may affect the spinal network and modulate the control of movement. Herein, we assessed the neuro-modulatory effect of sex hormones on the polysynaptic spinal network by using a flexion reflex network as a model system. Twenty-four healthy eumenorrheic women (age 21–37 years) were tested every other day for one menstrual cycle. Serum estradiol and progesterone were acquired at the time of testing. The flexion reflex of the tibialis anterior was elicited by sending an innocuous electrical stimulus directly to the posterior tibial nerve or plantar cutaneous afferent. Analyses were performed for each menstrual cycle phase: the follicular phase and the luteal phase. Increases in estradiol or progesterone concentrations were not associated with reflex duration or root mean squared (RMS) amplitude in either the follicular or luteal phases. In the luteal phase, an increase in the estradiol concentration was associated with a longer latency of the reflex (b = 0.23, p = 0.038). The estradiol × progesterone interaction was found towards significance (b = −0.017, p = 0.081). These results highlight the potential synergistic effect of estradiol and progesterone and may provide indirect confirmatory evidence of the observed modulatory effect.

The natural variation in estrogen secretion throughout the female menstrual cycle impacts various organs, including estrogen receptor (ER)-expressed skeletal muscle. Many women commonly experience increased fatigue or reduced energy levels in the days leading up to and during menstruation when blood estrogen levels decline. Yet, it remains unclear whether endogenous 17β-estradiol, a major estrogen component, directly affects energy metabolism in skeletal muscle due to the intricate and fluctuating nature of female hormones. In this study, we employed 3D 31P FID-MRSI to investigate phosphoryl metabolites in the soleus muscle of a cohort of young females (average age: 28 ± 6 years, n = 7) during the early follicular (EF) and peri-ovulation (PO) phases when their blood 17β-estradiol levels differ significantly (EF: 28 ± 18 pg/mL vs. PO: 71 ± 30 pg/mL, p < 0.05), while the levels of other potentially interfering hormones remain relatively invariant. Our findings reveal a reduction in ATP-referenced phosphocreatine (PCr) levels in the EF phase compared to the PO phase for all participants (5.4 ± 4.3%). Furthermore, we observe a linear correlation between muscle PCr levels and blood 17β-estradiol concentrations (r = 0.64, p = 0.014). Conversely, inorganic phosphate Pi and phospholipid metabolite GPC levels remain independent of 17β-estradiol but display a high correlation between EF and PO phases (p = 0.015 for Pi and p = 0.0008 for GPC).The robust association we have identified between ATP-referenced PCr and 17β-estradiol suggests that 17β-estradiol plays a modulatory role in the energy metabolism of skeletal muscle.

Estimating the state of tract-specific inputs to spinal motoneurons is critical to understanding movement deficits induced by neurological injury and potential pathways to recovery but remains challenging in humans. In this study, we explored the capability of trans-spinal magnetic stimulation (TSMS) to modulate distal reflex circuits in young adults. TSMS was applied over the thoracic spine to condition soleus H-reflexes involving sacral-level motoneurons. Three TSMS intensities below the motor threshold were applied at interstimulus intervals (ISIs) between 2 and 20 ms relative to peripheral nerve stimulation (PNS). Although low-intensity TSMS yielded no changes in H-reflexes across ISIs, the two higher stimulus intensities yielded two phases of H-reflex inhibition: a relatively long-lasting period at 2- to 9-ms ISIs, and a short phase at 11- to 12-ms ISIs. H-reflex inhibition at 2-ms ISI was uniquely dependent on TSMS intensity. To identify the candidate neural pathways contributing to H-reflex suppression, we constructed a tract-specific conduction time estimation model. Based upon our model, H-reflex inhibition at 11- to 12-ms ISIs is likely a manifestation of orthodromic transmission along the lateral reticulospinal tract. In contrast, the inhibition at 2-ms ISI likely reflects orthodromic transmission along sensory fibers with activation reaching the brain, before descending along motor tracts. Multiple pathways may contribute to H-reflex modulation between 4- and 9-ms ISIs, orthodromic transmission along sensorimotor tracts, and antidromic transmission of multiple motor tracts. Our findings suggest that noninvasive TSMS can influence motoneuron excitability at distal segments and that the contribution of specific tracts to motoneuron excitability may be distinguishable based on conduction velocities.