Stimulation Period Research Articles

Paired associative stimulation improves motor function in the upper extremity in chronic incomplete spinal cord injury: a corroborative study.

To corroborate findings suggesting that spinally targeted paired associative stimulation improves upper extremity motor function in chronic incomplete spinal cord injury. Prospective interventional study. Five adults with chronic tetraplegia. Participants received paired associative stimulation, combining peripheral nerve stimulation and navigated transcranial magnetic stimulation towards 1 arm (16 1-h sessions during 4 consecutive weeks, targeting the 3 large nerves). Manual muscle testing (MMT) was performed in 23 muscles in each arm, at 3 time points (pre-stimulation, t0; the week following the stimulation period, t1; and 4-5 weeks post-stimulation, t2). Additionally, grip strength and changes in the Canadian Occupational Performance Measure were assessed. The mean improvement in manual muscle testing scores in the targeted extremity was +0.49 at t1 (p = 0.078) and +0.55 at t2 (p = 0.062). Grip strength in the stimulated extremity increased by 3.2 kg at t1 and 3.4 kg at t2, and in the non-targeted extremity by 2.2 and 3.6 kg, respectively. Performance and satisfaction increased by 2.1/2.4 points at t1, and by 2.0/1.9 points at t2. Paired associative stimulation improved motor function: at the group level, MMT of the stimulated hand (p = 0.06) and non-stimulated hand (p = 0.04). Most participants achieved clinically relevant improvement. Thus, the results corroborate prior studies. The method may complement conventional rehabilitation for improving upper extremity function in incomplete tetraplegia.

Activation-Induced Marker Assay to Identify and Isolate HCV-Specific T Cells for Single-Cell RNA-Seq Analysis.

Identification and isolation of antigen-specific T cells for downstream transcriptomic analysis is key for various immunological studies. Traditional methods using major histocompatibility complex (MHC) multimers are limited by the number of predefined immunodominant epitopes and MHC matching of the study subjects. Activation-induced markers (AIM) enable highly sensitive detection of rare antigen-specific T cells irrespective of the availability of MHC multimers. Herein, we have developed an AIM assay for the detection, sorting and subsequent single-cell RNA sequencing (scRNA-seq) analysis of hepatitis C virus (HCV)-specific T cells. We examined different combinations of the activation markers CD69, CD40L, OX40, and 4-1BB at 6, 9, 18 and 24 h post stimulation with HCV peptide pools. AIM+ CD4 T cells exhibited upregulation of CD69 and CD40L as early as 6 h post-stimulation, while OX40 and 4-1BB expression was delayed until 18 h. AIM+ CD8 T cells were characterized by the coexpression of CD69 and 4-1BB at 18 h, while the expression of CD40L and OX40 remained low throughout the stimulation period. AIM+ CD4 and CD8 T cells were successfully sorted and processed for scRNA-seq analysis examining gene expression and T cell receptor (TCR) usage. scRNA-seq analysis from this one subject revealed that AIM+ CD4 T (CD69+ CD40L+) cells predominantly represented Tfh, Th1, and Th17 profiles, whereas AIM+ CD8 T (CD69+ 4-1BB+) cells primarily exhibited effector and effector memory profiles. TCR analysis identified 1023 and 160 unique clonotypes within AIM+ CD4 and CD8 T cells, respectively. In conclusion, this approach offers highly sensitive detection of HCV-specific T cells that can be applied for cohort studies, thus facilitating the identification of specific gene signatures associated with infection outcome and vaccination.

Physiological costs of warning: Defensive hissing increases metabolic rate and evaporative water loss in a venomous snake

To minimize predation risk and the cost of confronting predators, prey have developed a range of defensive strategies and warning signals. Although advantageous, defensive warnings may also induce physiological and energy costs to the emitter. Ventilatory sounds (hissing) are the most distributed warning sound in vertebrates. Because they involve the respiratory apparatus, defensive hissing may substantially increase evaporative water loss. Herein, we examined the determinants of hissing as well as its physiological costs in a medium-sized venomous snake, the long-nosed viper (Vipera ammodytes). We first used a neutral arena and applied standardized stimulation to measure the occurrence and acoustic characteristics of warning hissing. Then, we used open-flow respirometry to quantify changes in respiratory gas exchanges (oxygen consumption and evaporative water loss) during defensive responses. We demonstrated that males are more likely to engage in sound warnings when stimulated. Expirations generated the strongest signals compared to inspiration but did not differ between sexes. We found that defensive hissing dramatically increased average metabolic rate and evaporative water loss during the 10-minute stimulation period, and this effect was more pronounced in males. Metabolic rates and evaporative water loss were closely related to the duration of hissing. Overall, our results indicate that respiratory-based warning sounds induce significant physiological costs and may alter water balance. The higher responsiveness in males than females likely reflects sexually selective pressure (higher mobility for mate acquisition) and enhanced risk exposure.

Simultaneous modulation of pulse charge and burst period elicits two differentiable referred sensations

Objective.To investigate the feasibility of delivering multidimensional feedback using a single channel of peripheral nerve stimulation by complementing intensity percepts with flutter frequency percepts controlled by burst period modulation.Approach.Two dimensions of a distally referred sensation were provided simultaneously: intensity was conveyed by the modulation of the pulse charge rate inside short discrete periods of stimulation referred to as bursts and frequency was conveyed by the modulation of the period between bursts. For this approach to be feasible, intensity percepts must be perceived independently of frequency percepts. Two experiments investigated these interactions. A series of two alternative forced choice tasks (2AFC) were used to investigate burst period modulation's role in intensity discernibility. Magnitude estimation tasks were used to determine any interactions in the gradation between the frequency and intensity percepts.Main results.The 2AFC revealed that burst periods can be individually differentiated as a gradable frequency percept in peripheral nerve stimulation. Participants could correctly rate a perceptual scale of intensity and frequency regardless of the value of the second, but the dependence of frequency differentiability on charge rate indicates that frequency was harder to detect with weaker intensity percepts. The same was not observed in intensity differentiability as the length of burst periods did not significantly alter intensity differentiation. These results suggest multidimensional encoding is a promising approach for increasing information throughput in sensory feedback systems if intensity ranges are selected properly.Significance.This study offers valuable insights into haptic feedback through the peripheral nervous system and demonstrates an encoding approach for neural stimulation that may offer enhanced information transfer in virtual reality applications and sensory-enabled prosthetic systems. This multidimensional encoding strategy for sensory feedback may open new avenues for enriched control capabilities.

Efficient generation of liver sinusoidal endothelial-like cells secreting coagulation factor VIII from human induced pluripotent stem cells

Liver sinusoidal endothelial cells (LSECs) and LSEC progenitor cells (LPCs) derived from human pluripotent stem cells (PSCs) are expected as valuable cell sources for the development of cell therapy for hemophilia A, a congenital deficiency of coagulation factor VIII (FVIII), as LSECs are responsible for FVIII production. However, there is room for improvement in the efficiency of LSECs and LPCs differentiation from PSCs. In this study, we sought to optimize the method of mesoderm differentiation induction, the initial step of LSEC differentiation from PSCs, to efficiently induce LSEC-like cells capable of secreting FVIII from human induced pluripotent stem cells (iPSCs). Following optimization of the concentration and stimulation period of CHIR99021 (glycogen synthase kinase 3β inhibitor), bone morphogenetic protein 4, fibroblast growth factor 2, and Activin A in the mesoderm induction step, approximately 65% and 54% of cells differentiated into LPCs and LSEC-like cells, respectively. Furthermore, we observed substantial FVIII protein secretion from LSEC-like cells in vitro. In conclusion, we established an efficient method for obtaining LPCs and functional LSEC-like cells from iPSCs in vitro.

Visually induced involuntary arm, head, and torso movements

We explored in 75 s long trials the effects of visually induced self-rotation and displacement (SR&D) on the horizontally extended right arm of standing subjects (N = 12). A “tool condition” was included in which subjects held a long rod. The extent of arm movement was contingent on whether the arm was extended out Freely or Pointing at a briefly proprioceptively specified target position. The results were nearly identical when subjects held the rod. Subjects in the Free conditions showed significant unintentional arm deviations, averaging 55° in the direction opposite the induced illusory self-motion. Deviations in the Pointing conditions were on average a fifth of those in the Free condition. Deviations of head and torso positions also occurred in all conditions. Total arm and head deviations were the sum of deviations of the arm and head with respect to the torso and deviations of the torso with respect to space. Pointing subjects were able to detect and correct for arm and head deviations with respect to the torso but not for the arm and head deviations with respect to space due to deviations of the torso. In all conditions, arm, head, and torso deviations began before subjects experienced SR&D. We relate our findings to being an extension of the manual following response (MFR) mechanism to influence passive arm control and arm target maintenance as well. Visual-vestibular convergence at vestibular nuclei cells and multiple cortical movement related areas can explain our results, MFR results, and classical Pass Pointing. We distinguish two Phases in the induction of SR&D. In Phase 1, the visual stimulation period prior to SR&D onset, the arm, head, and torso deviations are first apparent, circa < 1 s after stimulus begins. They are augmented at the onset of Phase 2 that starts when SR&D is first sensed. In Phase 2, reaching movements first show curved paths that are compensatory for the Coriolis forces that would be generated on the reaching arm were subjects actually physically rotating. These movement deviations are in the opposite direction to the MFR and the arm, head, and torso deviations reported here. Our results have implications for vehicle control in environments that can induce illusory self motion and displacement.

Classical, spaced, or accelerated transcranial magnetic stimulation of motor cortex for treating neuropathic pain: A 3-arm parallel non-inferiority study

BackgroundRepetitive transcranial magnetic stimulation (rTMS) of the primary motor cortex (M1) at high frequency (HF) is an effective treatment of neuropathic pain. The classical HF-rTMS protocol (CHF-rTMS) includes a daily session for one week as an induction phase of treatment followed by more spaced sessions. Another type of protocol without an induction phase and based solely on spaced sessions of HF-rTMS (SHF-rTMS) has also been shown to produce neuropathic pain relief. However, CHF-rTMS and SHF-rTMS of M1 have never been compared regarding their analgesic potential. Another type of rTMS paradigm, called accelerated intermittent theta burst stimulation (ACC-iTBS), has recently been proposed for the treatment of depression, the other clinical condition for which HF-rTMS is proposed as an effective therapeutic strategy. ACC-iTBS combines a high number of pulses delivered in short sessions grouped into a few days of stimulation. This type of protocol has never been applied to M1 for the treatment of pain. Methods/DesignThe objective of this single-centre randomized study is to compare the efficacy of three different rTMS protocols for the treatment of chronic neuropathic pain: CHF-rTMS, SHF-rTMS, and ACC-iTBS. The CHF-rTMS will consists of 10 stimulation sessions, including 5 daily sessions of 10Hz-rTMS (3,000 pulses per session) over one week, then one session per week for 5 weeks, for a total of 30,000 pulses delivered in 10 stimulation days. The SHF-rTMS protocol will only include 4 sessions of 20Hz-rTMS (1,600 pulses per session), one every 15 days, for a total of 6,400 pulses delivered in 4 stimulation days. The ACC-iTBS protocol will comprise 5 sessions of iTBS (600 pulses per session) completed in half a day for 2 consecutive days, repeated 5 weeks later, for a total of 30,000 pulses delivered in 4 stimulation days. Thus, CHF-rTMS and ACC-iTBS protocols will share a higher total number of TMS pulses (30,000 pulses) compared to SHF-rTMS protocol (6,400 pulses), while CHF-rTMS protocol will include a higher number of stimulation days (10 days) compared to ACC-iTBS and SHF-rTMS protocols (4 days). In all protocols, the M1 target will be defined in the same way and stimulated at the same intensity using a navigated rTMS (nTMS) procedure. The evaluation will be based on clinical outcomes with various scales and questionnaires assessed every week, from two weeks before the 7-week period of therapeutic stimulation until 4 weeks after. Additionally, three sets of neurophysiological outcomes (resting-state electroencephalography (EEG), nTMS-EEG recordings, and short intracortical inhibition measurement with threshold tracking method) will be assessed the week before and after the 7-week period of therapeutic stimulation. DiscussionThis study will make it possible to compare the analgesic efficacy of the CHF-rTMS and SHF-rTMS protocols and to appraise that of the ACC-iTBS protocol for the first time. This study will also make it possible to determine the respective influence of the total number of pulses and days of stimulation delivered to M1 on the extent of pain relief. Thus, if their analgesic efficacy is not inferior to that of CHF-rTMS, SHF-rTMS and especially the new ACC-iTBS protocol could be an optimal compromise of a more easy-to-perform rTMS protocol for the treatment of patients with chronic neuropathic pain.

Noninvasive brain stimulation during EEG improves machine learning classification in chronic stroke.

In individuals with chronic stroke and hemiparesis, noninvasive brain stimulation (NIBS) may be used as an adjunct to therapy for improving motor recovery. Specific states of movement during motor recovery are more responsive to brain stimulation than others, thus a system that could auto-detect movement state would be useful in correctly identifying the most effective stimulation periods. The aim of this study was to compare the performance of different machine learning models in classifying movement periods during EEG recordings of hemiparetic individuals receiving noninvasive brain stimulation. We hypothesized that transcranial direct current stimulation, a form of NIBS, would modulate brain recordings correlating with movement state and improve classification accuracies above those receiving sham stimulation. Electroencephalogram data were obtained from 10 participants with chronic stroke and 11 healthy individuals performing a motor task while undergoing transcranial direct current stimulation. Eight traditional machine learning algorithms and five ensemble methods were used to classify two movement states (a hold posture and an arm reaching movement) before, during and after stimulation. To minimize compute times, preprocessing and feature extraction were limited to z-score normalization and power binning into five frequency bands (delta through gamma). Classification of disease state produced significantly higher accuracies in the stimulation (versus sham) group at 78.9% (versus 55.6%, p < 0.000002). We observed significantly higher accuracies when classifying stimulation state in the chronic stroke group (77.6%) relative to healthy controls (64.1%, p < 0.0095). In the chronic stroke cohort, classification of hold versus reach was highest during the stimulation period (75.2%) as opposed to the pre- and post-stimulation periods. Linear discriminant analysis, logistic regression, and decision tree algorithms classified movement state most accurately in participants with chronic stroke during the stimulation period (76.1%). For the ensemble methods, the highest classification accuracy for hold versus reach was achieved using low gamma frequency (30-50 Hz) as a feature (74.5%), although this result did not achieve statistical significance. Machine learning algorithms demonstrated sufficiently high movement state classification accuracy in participants with chronic stroke performing functional tasks during noninvasive brain stimulation. tDCS improved disease state and movement state classification in participants with chronic stroke.

Different Numbers of Conjunctive Stimuli Induce LTP or LTD in Mouse Cerebellar Purkinje Cell.

Long-term depression (LTD) of synaptic transmission at parallel fiber (PF)-Purkinje cell (PC) synapses plays an important role in cerebellum-related motor coordination and learning. LTD is induced by the conjunction of PF stimulation and climbing fiber (CF) stimulation or somatic PC depolarization, while long-term potentiation (LTP) is induced by PF stimulation alone. Therefore, it is considered that different types of stimulation induce different types of synaptic plasticity. However, we found that a small number of conjunctive stimulations (PF + somatic depolarization of PC) induced LTP, but did not induce LTD of a small size. This LTP was not associated with changes in paired-pulse ratio, suggesting postsynaptic origin. Additionally this LTP was dependent on nitric oxide. This LTP was also induced by a smaller number of physiological conjunctive PF and CF stimuli. These results suggested that a larger number or longer period of conjunctive stimulation is required to induce LTD by overcoming LTP. Ca2+ transients at the PC dendritic region was measured by calcium imaging during LTD-inducing conjunctive stimulation. Peak amplitude of Ca2+ transients increased gradually during repetitive conjunctive stimulation. Instantaneous peak amplitude was not different between the early phase and late phase, but the average amplitude was larger in the later phase than in the early phase. These results show that LTD overcomes LTP, and increased Ca2+ integration or a number of stimulations is required for LTD induction.

Electrical stimulation and wet ageing effects on the carcass and meat quality of Australian rangeland goats

Medium voltage electrical stimulation and wet ageing periods effects on the carcasses and meat quality of Australian rangeland goats were investigated. 64 rangeland goats were slaughtered as 2 equal consignments, wherein half were exposed to post-dressing electrical stimulation (300 V, 500 ms pulse width, 45 s) and half were not (control). Carcass and pH decline parameters and glycogen concentration were recorded. At 24 h post-mortem, fresh colour was measured and longissimus lumborum muscles (LL) removed. Within carcass, paired LL were halved and allocated to each of 4 ageing periods (1, 2, 4, or 21 d). There were no significant effects from electric stimulation on LL pH at 18 °C, LL temperature at pH 6, LL pH at 24 h post-mortem, semitendinosus pH at 24 h post-mortem, and LL glycogen concentration. There were no significant interactions between stimulation and ageing period on cooking loss, particle size, purge loss, sarcomere length, shear force, and ultimate pH. Independent to stimulation, shear force, particle size, and cooking values declined as ageing period increased. Purge loss was highest after 21 d of ageing. Colour stability was unaffected by ageing, although all CIE colour coordinates varied across the 74 h total retail display period. There were negative linear relationships found between LL glycogen concentrations and LL temperature at pH 6, ultimate pH, and sarcomere length. These findings suggest there to be little benefit to post-dressing electrical stimulation of rangeland goat carcasses, when applied independent to or in combination with ageing.

A sham-controlled, randomized trial of spinal cord stimulation for the treatment of pain in chronic pancreatitis.

Spinal cord stimulation (SCS) has emerged as a treatment option for patients with chronic pancreatitis (CP) who experience pain that does not respond to standard interventions. However, there is a lack of sham-controlled trials to support its efficacy. This randomized, double-blinded, sham-controlled, cross-over trial enrolled 16 CP patients with insufficient pain relief from standard therapies. Patients underwent high-frequency (1000 Hz) paraesthesia-free SCS or sham for two 10-day stimulation periods, separated by a 3-day washout period. The primary outcome was daily pain intensity registered in a pain diary based on a numeric rating scale (NRS). Secondary outcomes included various questionnaires. Quantitative sensory testing was used to probe the pain system before and after interventions. The average daily pain score on the NRS at baseline was 5.2 ± 1.9. After SCS, the pain score was 4.2 ± 2.1 compared to 4.3 ± 2.1 in the sham group (mean difference -0.1, 95% CI [-1.4 to 1.1]; P = 0.81). Similarly, no differences were observed between groups for the maximal daily pain score, secondary outcomes or quantitative sensory testing parameters. During an open-label, non-sham-controlled and non-blinded extension of the study, the average daily NRS was 5.2 ± 1.7 at baseline, 3.2 ± 1.8 at 3 months, 2.9 ± 1.9 at 6 months and 3.4 ± 2.2 at 12 months of follow-up (P = 0.001). In this first sham-controlled trial of SCS in painful CP, we did not find evidence of short-term pain relief with paraesthesia-free high-frequency (1000 Hz) stimulation. However, evaluation of the long-term effect by larger sham-controlled trials with long-term follow-up is warranted. In this first sham-controlled trial to apply high-frequency (1000 Hz) spinal cord stimulation in patients with visceral pain due to chronic pancreatitis, we did not find evidence for clinically relevant pain relief. Taken together with potential procedure-related complications, adverse effects and costs associated with spinal cord stimulation, our findings question its use for management of visceral pain.

Identifying SCS Trial Responders Immediately After Postoperative Programming with ECAP Dose-Controlled Closed-Loop Therapy.

Drawbacks of fixed-output spinal cord stimulation (SCS) screening trials may lead to compromised trial outcomes and poor predictability of long-term success. Evoked compound action potential (ECAP) dose-controlled closed-loop (CL) SCS allows objective confirmation of therapeutic neural activation and pulse-to-pulse stimulation adjustment. We report on the immediate patient-reported and neurophysiologic treatment response post-physiologic CL-SCS and feasibility of early SCS trial responder prediction. Patient-reported pain relief, functional improvement, and willingness to proceed to permanent implant were compared between the day of the trial procedure (Day 0) and end of trial (EOT) for 132 participants in the ECAP Study undergoing a trial stimulation period. ECAP-based neurophysiologic measurements from Day 0 and EOT were compared between responder groups. A high positive predictive value (PPV) was achieved with 98.4% (60/61) of patients successful on the Day 0 evaluation also responding at EOT. The false-positive rate (FPR) was 5.6% (1/18). ECAP-based neurophysiologic measures were not different between patients who passed all Day 0 success criteria ("Day 0 successes") and those who did not ("needed longer to evaluate the therapy"). However, at EOT, responders had higher therapeutic usage and dose levels compared to non-responders. The high PPV and low FPR of the Day 0 evaluation provide confidence in predicting trial outcomes as early as the day of the procedure. Day 0 trials may be beneficial for reducing patient burden and complication rates associated with extended trials. ECAP dose-controlled CL-SCS therapy may provide objective data and rapid-onset pain relief to improve prognostic ability of SCS trials in predicting outcomes. The ECAP Study is registered with ClinicalTrials.gov (NCT04319887).

P-594 Retrospective comparative study of follitropin delta vs follitropin alfa in progestin-primed ovarian stimulation (PPOS) protocol in IVF

Abstract Study question Could the individualized fixed-dose algorithm with follitropin delta show a similar pregnancy rate to conventional dosing with follitropin alfa in PPOS protocol? Summary answer Individualized fixed-dose algorithm with follitropin delta showed a similar pregnancy rate to follitropin alfa In PPOS protocol. What is known already Follitropin delta (REKOVELLE, Ferring Pharmaceuticals, Switzerland) is the first human cell line driven recombinant follicle-stimulating hormone (FSH) with an individualized fixed-dose algorithm based on serum AMH level and body weight. Follitropin alfa is a Chinese hamster ovarian cell driven recombinant FSH and its dosing is adjusted according to follicular response during the stimulation period. There was no significant difference between follitropin delta and follitropin alfa in terms of pregnancy rate under GnRH antagonist protocol. Study design, size, duration A retrospective analysis of 355 patients aged under 40 years who underwent ovarian stimulation in PPOS protocol using follitropin delta or follitropin alfa and on whom frozen-thawed single blastocyst transfer was conducted between August 2021 and November 2023 was performed. Participants/materials, setting, methods 193 cycles of women who used follitropin delta were compared to 162 cycles of women who used follitropin alfa. The following clinical outcomes on each treatment group were analyzed: chemical pregnancy rate (serum β hCG), clinical pregnancy rate (gestational sac), ongoing pregnancy rate(gestational sac and fetal heartbeat), serum E2 level at trigger day, cycle proportion of ≥ 10 oocytes retrieval, blastocyst formation rate, cycle proportion of ≥ 5 blastocysts and cycle proportion of ≥ 2 good quality blastocysts. Main results and the role of chance Baseline demographics in the follitropin delta group and the follitropin alfa group were not significantly different: age, 33.9 ± 3.5 years vs 34.5 ± 3.2 years; body weight, 53.5 ± 8.3 kg vs 53.8 ± 5.9 kg; number of previous IVF cycles, 0.2 ± 0.6 vs 0.3 ± 0.7; serum AMH level, 5.5 ± 2.9 ng/mL vs 5.3 ± 3.8 ng/mL, respectively. The chemical pregnancy rate (69.9% vs. 64.8%, P = 0.308), clinical pregnancy rate (58.0% vs. 56.2%, P = 0.748) and ongoing pregnancy rate (51.8% vs. 46.3%, P = 0.338) were comparable between the follitropin delta group and the follitropin alfa group. The individualized follitropin delta treatment resulted in a smaller cycle proportion of ≥ 10 oocytes retrieval (77.7% vs. 90.8%, P &amp;lt; 0.001) with lower serum E2 level at trigger day (3273 ± 1577 pg/mL vs. 3927 ± 1732 pg/mL, p &amp;lt; 0.001). However, the blastocyst formation rate was significantly higher in the follitropin delta group (75.5% vs. 68.7%, p &amp;lt; 0.001). Moreover, the cycle proportion of ≥ 5 blastocysts (71.0% vs. 76.4%, P = 0.286) and the cycle proportion of ≥ 2 good quality blastocysts (51.8% vs. 60.3%, P = 0.115) were comparable between the groups. Limitations, reasons for caution This is a non-controlled, retrospective study. Wider implications of the findings Follitropin delta in its individualized fixed-dose regimen could have the potential to decrease the risk of OHSS by minimizing excessive ovarian response, whereas a similar pregnancy rate to conventional dosing regimen could be expected due to the number of good quality blastocysts secured. Trial registration number not applicable

P-661 Comparative and descriptive molecular analysis of cumulus and luteinized mural granulosa cells for steroidogenic function in IVF patients

Abstract Study question Do cumulus granulosa cells (CCs) have steroidogenic function and gonadotropin response like luteinized mural granulosa cells (LMGCs)? Summary answer Yes. Both types of granulosa cells have similar expression levels for the steroidogenic enzymes and hCG/LH receptors. What is known already CCs and LMGCs have distinct roles in follicular development, oocyte growth and maturation, cumulus expansion, ovulation, and luteinization. CCs support oocytes, especially during meiotic resumption, ovulation, and fertilization. They exhibit specialized functions in response to hormones like LH and participate in bidirectional communication with the oocyte. Although LMGCs are primarily responsible for steroidogenic function and progesterone (P4) production, very limited data is available as to whether CC themselves express the steroidogenic enzymes and gonadotropin receptors and produce sex steroids at basal or after gonadotropin stimulation in humans. We aimed to address this issue in the current study. Study design, size, duration An in-vitro basic science study was designed to compare CCs and LGMCs in the context of vitality, steroidogenic capabilities (both in baseline and response to rec-hCG (10 IU/ml), and EGF pathway responses. The assessment was planned to compare hormonal assay in spent culture and transcriptional levels. The study’s duration encompassed the period of ovarian stimulation, oocyte retrieval, and subsequent cellular analysis. Participants/materials, setting, methods 24 IVF patients undergoing ovarian stimulation with progestin primed (PPOS) and GnRH antagonist protocols for different infertility etiologies were included. Ovarian stimulation and ovulation trigger were done using rec-FSH and rec-hCG, respectively. CCs were collected from the denudation plate. LMGCs were obtained from follicular aspirates during the OPU and cultured individually for each patient without pooling in culture plates. After overnight incubation, the cells were treated with rec-hCG overnight. RNA isolation was performed. Main results and the role of chance The expression of the steroidogenic enzymes (StAR, SCC, 17B-HSD, 3B-HSD, Aromatase), LH receptor and EGF-like amphiregulin (AREG) in CCs were comparable to LMGCs on quantitative RT-PCR analysis. In line with this finding, we found they produced similar amounts of estradiol (E2) and progesterone (P4) in culture. Furthermore, similar to LMGCs, hCG treatment of the CCs resulted in a significant increase in P4 output, indicative of the presence of functional LH receptors. When the culture period was extended to six days, CCs retained their vitality and steroidogenic function like LMGCs and produced detectable E2 and P4. Limitations, reasons for caution Caution should be exercised when interpreting our data as it is unclear if the parameters analyzed here vary according to the infertility etiology, stimulation protocol, mode of trigger and any underlying disease that concomitantly present. Wider implications of the findings Prominent steroidogenic activity of CCs might have other important implications for a successful pregnancy. Moreover, similar expression profiles of steroidogenic pathways and hCG/LH receptors between CCs and LMGCs indicate that CCs can also be used to understand better the luteal function and intricate crosstalk between in oocyte microenvironment. Trial registration number N/A

P-581 Optimizing efficiency and enhancing patient convenience in DuoStim cycles: A study exploring the continuous application of corifollitropin alpha

Abstract Study question Corifollitropin alfa in DuoStim cycles: is weekly continuous dosing a viable alternative to daily FSH administration for comparable oocyte yield? Summary answer Employing corifollitropin alfa on a weekly basis throughout a DuoStim cycle achieves comparable oocyte numbers to daily FSH while significantly reducing the number of injections. What is known already The DuoStim protocol is frequently employed in patients with poor response or low ovarian reserve. However, its utilization has recently been proposed in patients who require elective oocyte/embryo freezing with the aim of maximizing the number of gametes/embryos obtained in a menstrual cycle. While its popularity is growing, the prolonged stimulation period involves the administration of a high number of injectable medications adding to the treatment burden. Study design, size, duration A non-inferiority, paired-matched study was planned with a minimum sample size of 13 patients per group, based on a tolerated difference of ± 1 oocyte, SD: 1, 80% power and type 1 error: 5%. Study group (n = 30): patients undergoing DuoStim from November 2022–December 2023. The control group (n = 30) was selected from patients undergoing DuoStim in years 2021-2022 following a standard daily-FSH administration. Controls were 1-to-1 matched based on AMH (±0.4 pmol/l) and age. Participants/materials, setting, methods The study group received consecutive injections of 150µg corifollitropin alfa every eight days, along with uninterrupted oral administration of natural progesterone 200 mg/day (LH surge prevention) throughout the follicular and luteal phases. The control group employed a daily-FSH/flexible GnRH-antagonist protocol. GnRH-agonist trigger was used in the follicular phase, whereas either hCGr and/or GnRH-agonist triggering was allowed in the luteal phase. Main results and the role of chance Baseline characteristics: age (years), AMH (pmol/l), and BMI (Kg/m2) were comparable between control vs study group (39.7vs40.3, 6.1vs6.1, 24.4vs23.9; p0.05, p0.5 and p0.9, respectively). In terms of the primary objective: total COCs and MII yield (follicular + luteal phase); the standard daily-FSH protocol vs study group achieved a similar total oocyte and MII yield (10.9±5.5 vs 10.4±7.0, p0.6 and 8.7±4.8 vs 7.8±5.6, p0.4, respectively). Additionally, all secondary outcomes (fertilisation rates, number of blastocysts, days of stimulation and gonadotrophin cost) showed no significant differences between the groups. Notably, the study group experienced a remarkable reduction in the number of injections needed to complete a DuoStim cycle [3.8±1.1 vs 30.9±12.9, p &amp;lt; 0.001; MD -27.1±12.5, (95%CI, -49 to -8)]. Limitations, reasons for caution The secondary outcomes revealed comparable fertilisation rates and blastocyst numbers between the two groups. Nevertheless, it is important to interpret the laboratory results beyond oocyte collection cautiously, considering the smaller sample size (n = 23/group). Larger participant cohorts are necessary to validate these findings and mitigate the risk of type II errors. Wider implications of the findings This protocol involving long-acting FSH in combination with natural progesterone offers a viable alternative for patients undergoing DuoStim. It enables comparable results in terms of egg retrieval while minimizing the need for injectables, without increasing the treatment duration or costs. Consequently, it alleviates emotional and therapeutic burdens experienced by patients. Trial registration number NCT05815719

Gaze and attention: mechanisms underlying the therapeutic effect of optokinetic stimulation in spatial neglect

Left smooth pursuit eye movement training in response to large-field visual motion (optokinetic stimulation) has become a promising rehabilitation method in left spatial inattention or neglect. The mechanisms underlying the therapeutic effect, however, remain unknown. During optokinetic stimulation, there is an error in visual localization ahead of the line of sight. This could indicate a change in the brain’s estimate of one’s own direction of gaze. We hypothesized that optokinetic stimulation changes the brain’s estimate of gaze. Because this estimate is critical for coding the locus of attention in the visual space relative to the body and across sensory modalities, its change might underlie the change in spatial attention. Here, we report that in healthy participants optokinetic stimulation causes not only a directional bias in the proprioceptive signal from the extraocular muscles, but also a corresponding shift of the locus of attention. Both changes outlasted the period of stimulation. This result forms a step in investigating a causal link between the adaptation in the sensorimotor gaze signals and the recovery in spatial neglect.

Transcutaneous vagal nerve stimulation during lower body negative pressure

BackgroundPara-sympathetic vagal activation has profound influence on heart rate and other cardiovascular parameters. We tested the hypothesis that transcutaneous Vagal Nerve Stimulation (tVNS) through the auricular branch of the vagus nerve would attenuate the normal sympathetic response to central blood volume reduction by lower body negative pressure (LBNP). Method10 healthy volunteers (6 female; age 21 ± 2 years; weight 62 ± 13 kg; height 167 ± 12 cm) were included in this cross-over design trial. After 15 min rest in supine position, subjects underwent three 15-min periods of 30 mmHg LBNP intervention with and without cyclic tVNS stimulation. Continuous cardiovascular parameters (Nexfin) were recorded. ResultsOverall tVNS did not convincingly attenuate sympathetic response to central hypovolemia. Deactivation of the tVNS during LBNP resulted in increased MAP at 2.3 ± 0.5 mmHg (P < 0.001). Comparing the cyclic actual active stimulation periods to periods with pause during tVNS intervention showed a decrease in HR by 72.9 ± 11.2 to 70.2 ± 11.6 bpm (mean ± SD; P < 0.05), and concomitant increases in SV (86.0 ± 12.1 to 87.2 ± 12.6 mL; P < 0.05), MAP (82.9 ± 6.3 to 84.0 ± 6.2 mmHg; P < 0.05) and TPR (1116.0 ± 111.1 to 1153 ± 104.8 dyn*s/cm5; P < 0.05). ConclusiontVNS in 30 s cycles during LBNP can selectively attenuate HR, prompting a compensatory augmented sympathetic response. It would appear the method used in this study at least, has an isolated cardiac inhibitory effect probably mediated by augmented vagal activity on the sinoatrial or atrio-ventricular node, possibly in combination with reduced activity in the sympathetic cardiac nerve.

No Consistent Antidepressant Effects of Deep Brain Stimulation of the Bed Nucleus of the Stria Terminalis.

Applying deep brain stimulation (DBS) to several brain regions has been investigated in attempts to treat highly treatment-resistant depression, with variable results. Our initial pilot data suggested that the bed nucleus of the stria terminalis (BNST) could be a promising therapeutic target. The aim of this study was to gather blinded data exploring the efficacy of applying DBS to the BNST in patients with highly refractory depression. Eight patients with chronic severe treatment-resistant depression underwent DBS to the BNST. A randomised, double-blind crossover study design with fixed stimulation parameters was followed and followed by a period of open-label stimulation. During the double-blind crossover phase, no consistent antidepressant effects were seen with any of the four stimulation parameters applied, and no patients achieved response or remission criteria during the blinded crossover phase or during a subsequent period of three months of blinded stimulation. Stimulation-related side effects, especially agitation, were reported by a number of patients and were reversible with adjustment of the stimulation parameters. The results of this study do not support the application of DBS to the BNST in patients with highly resistant depression or ongoing research utilising stimulation at this brain site. The blocked randomised study design utilising fixed stimulation parameters was poorly tolerated by the participants and does not appear suitable for assessing the efficacy of DBS at this location.

Wired for Success: Probing the Effect of Tissue-Engineered Neural Interface Substrates on Cell Viability.

This study investigates the electrochemical behavior of GelMA-based hydrogels and their interactions with PC12 neural cells under electrical stimulation in the presence of conducting substrates. Focusing on indium tin oxide (ITO), platinum, and gold mylar substrates supporting conductive scaffolds composed of hydrogel, graphene oxide, and gold nanorods, we explored how the substrate materials affect scaffold conductivity and cell viability. We examined the impact of an optimized electrical stimulation protocol on the PC12 cell viability. According to our findings, substrate selection significantly influences conductive hydrogel behavior, affecting cell viability and proliferation as a result. In particular, the ITO substrates were found to provide the best support for cell viability with an average of at least three times higher metabolic activity compared to platinum and gold mylar substrates over a 7 day stimulation period. The study offers new insights into substrate selection as a platform for neural cell stimulation and underscores the critical role of substrate materials in optimizing the efficacy of neural interfaces for biomedical applications. In addition to extending existing work, this study provides a robust platform for future explorations aimed at tailoring the full potential of tissue-engineered neural interfaces.

Super long ultraviolet-A persistent luminescence and photo-stimulated luminescence from CaMgGeO4: Bi3+

Here, we have developed a novel ultra long ultraviolet-A afterglow material CaMgGeO4: Bi3+, which shows more than 24 h UVA afterglow emission peaked at 346 nm. The crystal structure shows that the prepared sample is a pure phase with an orthogonal system of Pmnb space group. The composition of valence band and conduction band was analyzed by the density of states, which reveals that the bandgap of the CaMgGeO4 matrix is 3.69 eV. The photoluminescence spectrum of the sample consists of a broadband in the 320–400 nm UVA region, which belongs to the 3P1 → 1S0 electron transition of Bi3+. More importantly, under the excitation of ultraviolet lamp for 3 min, CaMgGeO4: Bi3+ exhibited an UVA ultra long persistent luminescence (LPL) with a duration of more than 24 h, which afterglow emission intensity still reaches 1500 cps and is about 6 times of the background intensity. In addition, the UVA emission of CaMgGeO4: Bi3+ after 13 h day can be repeatedly by short periods of low-energy light sources stimulation. The thermoluminescence analysis indicates that the presence of shallow and deep traps is responsible for excellent UVA-LPL and photo-stimulate luminescence (PSL). The excellent LPL and PSL properties can prompt CaMgGeO4: Bi3+ phosphor to be widely used in the field of photodynamic therapy.