Shorter Duration Of Stimulation Research Articles

Early Chromatin Remodeling Events in Acutely Stimulated CD8+ T Cells.

T cells undergo extensive chromatin remodeling over several days following stimulation through the T cell receptor. However, the kinetics and gene loci targeted by early remodeling events within the first 24 hours of T cell priming to orchestrate effector differentiation have not been well described. We identified that chromatin accessibility is rapidly and extensively remodeled within 1 hour of stimulation of naïve CD8+ T cells, leading to increased global chromatin accessibility at many effector T cell-associated genes that are enriched for AP-1, early growth response (EGR), and nuclear factor of activated T cells (NFAT) binding sites, but this short duration of stimulation is insufficient for commitment to clonal expansion in vivo. Sustained 24-hour stimulation led to further chromatin remodeling and was sufficient to enable clonal expansion. These data suggest that the duration of antigen receptor signaling is intimately coupled to chromatin remodeling and activation of genes involved in effector cell differentiation and highlight a potential mechanism that helps CD8+ T cells discriminate between foreign- and self-antigens.

CLINICAL RESULTS OF INFERTILITY TREATMENT IN PATIENTS WITH A COMORBIDITY OF UTERINE LEIOMYOMA AND CHRONIC ENDOMETRITIS WITH OPTIMIZATION OF CRYOPTOCOLS FOR EMBRYO TRANSFER PREPARATION

Infertility is a major medical and social issue. In vitro fertilization is an advanced ART technique that helps to successfully cure infertility. Quality preparation for an IVF protocol and the subsequent pregnancy is important and should include treatment of concomitant pathology, evaluation of the condition of the uterine cavity and its optimal preparation in order to prevent reproductive failures and early obstetric losses. This study assessed the effectiveness of optimized cryoprotocols for preparation for embryo transfer in patients with infertility and the comorbid course of uterine leiomyoma and chronic endometritis. We performed clinical examination and comparative analysis of anamnestic data of 100 women with infertility and comorbid uterine leiomyoma and chronic endometritis. The patients were divided into two groups depending on the proposed treatment scheme and the outcomes were compared to a control group. In all groups, stimulation of superovulation was carried out following a short protocol with a GnRH antagonist. We determined that patients with infertility comorbid with uterine leiomyoma and chronic endometritis, when compared to the control group patients, had a history of early menarche, early onset of sexual life, a greater number of sexual partners, a greater incidence of urogenital infections, and of performed intrauterine surgical procedures. The patients in the modified treatment group received a lower mean total dose of gonadotropins, had a shorter duration of stimulation, and fewer days of anti-GnRH administration compared to the traditional treatment group. The patients in the modified treatment group also had a higher frequency of clinical pregnancy and delivery compared to those received conventional treatment. An appropriate anti-inflammatory and antibiotic therapy and comprehensive preparation of the uterine cavity for embryo transfer are key factors in the success of IVF protocols.

Effectiveness of letrozole in pituitary downregulated normogonadotrophic young women with an initial poor response.

It has been reported that 10 to 15% of young normogonadotrophic women show suboptimal response to standard long protocols. Letrozole (LE), an aromatase inhibitor, was shown to improve ovarian sensitivity to follicle stimulating hormone (FSH) and follicular response to gonadotrophin treatment in poor ovarian response patients. We reasoned that it might be possible to utilize LE in young normogonadotrophic patients with unexpected hypo-response in standard gonadotropin-releasing hormone agonist long protocol. A total of 652 patients defined as normogonadotrophic patients with unexpected hypo-response were divided into 2 groups, the +LE group and the +Gn group. +LE group: A fixed daily dose of 2.5 mg of LE was added on day 8 of stimulation. +Gn group: A fixed daily dose of 75 U of human menopausal gonadotrophin was added on day 8 of stimulation. The primary outcome measures were the number of oocytes obtained, fertilization rate, days of stimulation, and total FSH dosage. The secondary outcome measures were the implantation rate and ongoing pregnancy rate. There were no significant differences in the clinical and hormonal characteristics between the 2 groups. A shorter duration of stimulation and a lower dosage of recombinant FSH consumption on the day of human chorionic gonadotropin administration were all observed in the +LE group. Patients who received LE therapy showed a higher number of oocytes obtained and significantly higher fertilization rates. The implantation rate and ongoing pregnancy rate were comparable in both groups. LE significantly improves the number of oocytes obtained in patients with suboptimal response to standard gonadotropin-releasing hormone agonist long protocol.

Identification of common genetic polymorphisms associated with down-regulated gonadotropin levels in an exome-wide association study

To investigate whether common genetic polymorphisms are associated with gonadotropin levels after down-regulation with daily gonadotropin-releasing hormone agonist and whether the polymorphisms of candidate variants influence the ovarian response to exogenous gonadotropins. Genetic association study. University-affiliated invitro fertilization center. Subjects enrolled in an exploratory exome-wide association study (n = 862), a replication exome-wide association study (n= 86), and a classifier validation study (n = 148) were recruited from September 2016 to October 2018, September 2019 to September 2020, and January 2021 to December 2021, respectively. The included patients were aged ≤40 years and had a basal follicle-stimulating hormone (FSH) ≤12 IU/L. All participants received a luteal phase down-regulation long protocol. Genome DNA was extracted from the peripheral blood leukocytes. For the exploratory and replication cohorts, exome sequencing was conducted on a HiSeq 2500 sequencing platform. The multiplex polymerase chain reaction amplification technique and next-generation sequencing also were performed in the exploratory and replication cohorts. For the samples of the validation cohort, Sanger sequencing was performed. The primary endpoint was the gonadotropin levels after down-regulation, and the secondary endpoints were hormone levels and follicle diameters during stimulation, the total dose of FSH, duration of FSH stimulation, number of oocytes retrieved, and clinical pregnancy rate. In the exploratory cohort, we identified that FSHB rs6169 (P=2.71 × 10-24) and its single-nucleotide polymorphisms in high linkage disequilibrium were associated with the down-regulated FSH level. The same locus was confirmed in the replication cohort. Women carrying the C allele of FSHB rs6169 exhibited higher average estradiol level during stimulation (P=6.82 × 10-5), shorter duration of stimulation, and less amount of exogenous FSH (Pduration=0.0002; Pdose=0.0024). In the independent validation set, adding rs6169 genotypes into the prediction model for FSH level after down-regulation enhanced the area under the curve from 0.560 to 0.712 in a logistic regression model, and increased prediction accuracy by 41.05% when a support vector machine classifier was applied. The C allele of FSHB rs6169 is a susceptibility site for the relatively high level of FSH after down-regulation, which may be associated with increased ovarian FSH sensitivity.

Effect of basal serum testosterone level on the ovarian response and the cumulative live birth rate in infertile women undergoing in vitro fertilization.

To evaluate the effect of basal serum testosterone levels on the ovarian response and the cumulative live birth rate of infertile women undergoing in vitro fertilization (IVF). It is a retrospective study in a university-affiliated assisted reproduction center in Hong Kong. Infertile women undergoing the first IVF cycle in the center between December 2012 and November 2016 with archived serum samples and available information on cumulative live birth were included for the analysis. A total of 1122 women were included for analysis. The median basal serum testosterone level was 0.53 (25-75th percentile: 0.40-0.67) nmol/L. Women with higher basal serum testosterone levels required a lower total dosage of gonadotrophin and a shorter duration of stimulation and had more oocytes retrieved. The cumulative live birth rates did not differ among women with serum testosterone levels in the four quartiles. Basal serum testosterone level was not a significant independent predictor of the cumulative live birth after adjusted for the women's age and number of normally fertilized oocytes in a binary logistic regression. The areas under the receiver operative characteristics (ROC) curves in predicting low or high ovarian response and the cumulative live birth were all below 0.6. Higher basal serum testosterone levels were associated with a better ovarian response but had no effect on the cumulative live birth rate of infertile women undergoing IVF.

P 2 Effects of deep brain stimulation (DBS) in the entopeduncular nucleus (EPN) in dystonic dtsz hamsters

Introduction: DBS of the globus pallidus internus (GPi; entopeduncular nucleus, EPN, in rodents) has become important for the treatment of generalized dystonia. There is evidence that striatal dysfunctions cause a disturbed thalamocortical inhibition via altered neuronal activity in the GPi/EPN. However, the pathophysiology as well as the mechanisms of DBS are largely unknown, which hampers the detection of drug targets. In our project of the CRC “Electrically active implants” we aim to elucidate mechanisms of DBS in animal models of dystonia. We recently found that short-term (3 h) EPN-DBS with 130 Hz (50 µA, 60 µs) improves paroxysmal dystonia in the dt sz mutant hamster 1 and reduces spontaneous excitatory cortico-striatal activity in brain slices of this model 2 , indicating fast effects in synaptic plasticity. In the present study, we therefore examined whether short-term DBS leads to changes of c-Fos, an immunohistochemical marker of neuronal activity, in EPN-related network regions. Materials & Methods: For immunohistochemistry we used brains of stimulated dt s z hamsters, in which DBS had improved dystonia, as well as sham-stimulated and naïve animals and performed cell counting and fluorescence intensity measurements of c-Fos within the basal ganglia network. In addition, we performed double labeling of c-Fos with GAD67 and with parvalbumin (PV), respectively, in brain slices of the same animals. Results: After DBS vs. sham, c-Fos + around the electrode was increased. Unexpectedly, c-Fos + cells were decreased in deep cerebellar nuclei (DCN) after DBS, but no changes became evident within the whole EPN, habenula, ventromedial thalamus, cortex and striatum. Cell counting of c-Fos activated GAD67 + as well as activated PV + cells showed no differences between the groups in motor cortex and striatum. Conclusion: With regard to recent electrophysiological data, we expected c-Fos changes especially in cortical GABAergic neurons after DBS in dt sz hamsters. The lack of changes within the basal ganglia network could be related to the short duration of stimulation or the time interval between stimulation and c-Fos staining. Ongoing long-term EPN stimulations, now feasible by a new fully implantable stimulator 3 , probably lead to more pronounced effects within the network and will be useful to verify the DCN effects. Funded by the DFG within the Collaborative Research Centre 1270 ELAINE.

A CNN-based multi-target fast classification method for AR-SSVEP

Because an augmented-reality-based brain-computer interface (AR-BCI) is easily disturbed by external factors, the traditional electroencephalograph (EEG) classification algorithms fail to meet the real-time processing requirements with a large number of stimulus targets or in a real environment. We propose a multi-target fast classification method for augmented-reality-based steady-state visual evoked potential (AR-SSVEP), using a convolutional neural network (CNN). To explore the availability and accuracy of high-efficiency multi-target classification methods in AR-SSVEP with a short stimulation duration, a similar stimulus layout was used for a computer screen (PC) and an optical see-through head-mounted display (OST-HMD) device (HoloLens). The experiment included nine flicker stimuli of different frequencies, and a multi-target fast classification method based on a CNN was constructed to complete nine classification tasks, for which the average accuracy of AR-BCI in our CNN model at 0.5- and 1-s stimulus duration was 67.93% and 80.83%, respectively. These results verified the efficacy of the proposed model for processing multi-target classification in AR-BCI.

The ultra-long study: a randomized controlled trial evaluating long-term GnRH downregulation prior to ART in women with endometriosis.

Does ultra-long downregulation with a GnRH agonist (triptorelin depot) in previously operated patients with endometriosis improve the rate of clinical pregnancy with positive fetal heart beat (CPHB) in the subsequent initiated fresh ART cycle? Ultra-long downregulation with a GnRH agonist prior to ART did not improve the rate of CPHB in the subsequent fresh ART cycle in previously completely operated patients but the trial was underpowered due to early termination. Administration of GnRH agonists for a period of 3-6 months prior to ART in women with endometriosis may increase the odds of clinical pregnancy. However, the quality of the studies on which this statement is based is questionable, so these findings need confirmation. A controlled, randomized, open label trial was performed between 1 June 2013 and 31 December 2016 (start and end of recruitment, respectively). Patients with prior complete laparoscopic treatment of any type or stage of endometriosis and an indication for ART were randomized (by a computer-generated allocation sequence) into two groups: the control group underwent ART stimulation in a classical long agonist protocol using preparation with oral contraceptives, the ultra-long group first underwent at least 3 months downregulation followed by a long agonist protocol for ART stimulation. The sample size was calculated to detect a superiority of the ultra-long downregulation protocol, based on the hypothesis that baseline CPHB rate in the control group of 20% would increase to 40% in the ultra-long group. For a power of 20% at a significance level of 5%, based on two-sided testing, including 5% of patients lost to follow-up, the necessary sample size was 172 patients (86 per group). This trial was conducted at the Leuven University Fertility Center, a tertiary care center for endometriosis and infertility, and a total of 42 patients were randomized (21 in the control group and 21 in the ultra-long group). Baseline characteristics were similar in both groups. The primary outcome studied-CPHB after the initiated ART treatment-did not differ and was 25% (5/20) in the control group, and 20% (4/20) in the ultra-long group (P > 0.999; relative risk (RR) 1.25, 95% CI 0.41-3.88). Cumulative (fresh + associated frozen) CPHB rates were also similar in the control versus ultra-long group (8/20, 40% vs 6/20, 30%, P = 0.7411; RR = 1.33, 95% CI 0.57-3.19). When other secondary outcomes were compared with the ultra-long group, patients from the control group had a shorter duration of stimulation (mean 11.8 days (SD ± 2.4) versus 13.2 days (SD ± 1.5), P = 0.0373), a lower total dose of gonadotrophins used (mean 1793 IU/d (SD ± 787) vs 2329 (SD ± 680), P = 0.0154), and a higher serum estradiol concentration (ng/ml) at the end of ovarian stimulation on the day of ovulation triggering or cycle cancellation (mean1971 (SD ± 1495) vs 929 (± 548); P = 0.0326), suggesting a better ovarian response in the control group. Due to a strong patient preference, nearly exclusively against ultra-long downregulation (even though patients were thoroughly informed of the potential benefits), the targeted sample size could not be achieved and the trial was stopped prematurely. Conditional power analysis revealed that the probability of confirming the study hypothesis if the study were completed would be low. We hypothesize that in patients with prior complete surgical treatment of endometriosis, the ultra-long protocol does not enhance ART-CPHB rates. Patient's concerns and preferences regarding possible side-effects, and delay of ART treatment start with the ultra-long protocol should be taken into account when considering this type of treatment in women with endometriosis. C.T. was during 2 years funded by a grant from the Clinical research Foundation of UZ Leuven (KOF) and during 2 years by the Research Foundation-Flanders (FWO grant number: 1700816N). C.T. reports grants from Clinical Research Foundation of the University Hospitals of Leuven (KOF), grants from Fund for Scientific Research Flanders (FWO), during the conduct of the study; grants, non-financial support and other from Merck SA, non-financial support and other from Gedeon Richter, non-financial support from Ferring Pharmaceuticals, outside the submitted work. T.D. is vice president and head of Global Medical Affairs Fertility, Research and Development, Merck KGaA, Darmstadt, Germany. He is also a professor in Reproductive Medicine and Biology at the Department of Development and Regeneration, Group Biomedical Sciences, KU Leuven (University of Leuven), Belgium and an adjunct professor at the Department of Obstetrics and Gynecology in the University of Yale, New Haven, USA. Neither his corporate role nor his academic roles represent a conflict of interest with respect to the work done by him for this study. A.C. reports personal fees from Merck S.p.A., outside the submitted work. The other co-authors have no conflict of interest. UZ Leuven trial registry SS55300, EudraCT number 2013-000993-32, clinicaltrials.gov NCT02400801. Registration for EudraCT on 1 March 2013. 4 September 2013.

P–325 The impact of letrozole on endometrial thickness in IVF cycles

Abstract Study question Does daily administration of letrozole during IVF stimulation affect endometrial thickness ? Summary answer Patients treated with letrozole during fresh IVF cycles had a thinner endometrium on the day of trigger compared to patients who did not receive letrozole. What is known already Letrozole supplementation is commonly used during fertility preservation for breast cancer patients to reduce peak estrogen levels with no adverse effects on embryo outcomes. Studies in poor responders have found that letrozole use resulted in a shorter duration of stimulation and a lower total dose of gonadotropin, with no detrimental effect on IVF outcomes. In normal responders, studies have shown an increase in blastocysts obtained, but have not yet shown an increase in clinical pregnancy rates. There is concern that when a fresh embryo transfer is planned letrozole use may negatively affect endometrial thickness and subsequently diminish pregnancy rates. Study design, size, duration In a retrospective cohort study between January 2009 and June 2019 at a single academic fertility center, we compared the endometrial thickness in 97 cancer patients who underwent IVF-fertility preservation with daily letrozole use to 158 cancer patients who underwent IVF-fertility preservation without letrozole. Participants/materials, setting, methods All women diagnosed with cancer were referred for fertility preservation prior to gonadotoxic treatment exposure and were less than 40 years old at the time of oocyte retrieval. All patients who received letrozole started on day one of stimulation and continued until the day of oocyte retrieval. The primary outcome was endometrial thickness on the day of trigger. The secondary outcomes were number of oocytes retrieved, number of MII retrieved, and maximal estradiol level. Main results and the role of chance During the study period, 336 cancer patients underwent fertility preservation. Eighty-one patients were excluded; 50 because they had an intrauterine device or were on long term oral contraceptives and 31 because endometrial thickness was not documented. Of the remaining 255 patients, 86 had breast cancer, 95 had a hematological cancer and 74 had various other cancers. Ninety-seven cancer patients treated with letrozole were compared to 158 cancer patients who did not receive letrozole. Patients who received letrozole were significantly older (34 vs 28yrs, P < 0.0001). There were no significant differences in baseline characteristics such as BMI, AFC nor in the total duration for stimulation. Endometrial thickness on the day of trigger was significantly less in letrozole treated patients (8 vs 9mm, P < 0.003). There were no significant differences in total number of oocytes retrieved (12.5 vs 11, P = 0.126) nor in the number of mature oocytes (8 vs 8, P = 0.312). Patients in the letrozole group received a higher total gonadotropin dose (2680IU vs 1980IU, P = 0.016). The maximum estradiol level was significantly lower in patients treated with letrozole (1068 vs 3838ml/dl, P = <0.0001). A regression analysis showed that using letrozole during stimulation decreased the endometrial thickness by 0.81mm (CI –1.37 to –0.253, P = 0.005). Limitations, reasons for caution The retrospective nature of this study could have introduced selection and misinformation bias. We report on cancer patients where all oocytes or embryos were vitrified. Without fresh embryo transfer data, it is unclear if a thinner endometrium due to letrozole will effect the implantation or pregnancy rate. Wider implications of the findings: As the use of letrozole expands beyond cancer patients and poor responders, it is important to understand the impact on the endometrium. This study shows that letrozole reduces endometrial thickness. However, the effect on endometrial function remains unknown. Further study is needed before letrozole can be used with fresh transfers. Trial registration number 2020–6370

Effect of electrical stimulation on the production of cutaneous excreta by the earthworm Lumbricus terrestris L

Cutaneous excreta of earthworms are an increasing interest in many areas of scientific research. Our study aims to develop an electrical technique for stimulating the production of these cutaneous excretions more efficient and less harmful than that using chemical stimulation with petroleum ether. This study made it possible to define the optimal electrical parameters ensuring the obtaining of a maximum rate of excretion. These parameters are respectively: a voltage (U) of 20V, a pulse frequency (F) of 100 stimulations per second and a stimulation time (T) of 5 minutes. The comparative study of this new technique of electrical stimulation compared to chemical stimulation shows statistically very comparable excretion production rates (p<0,05). However, electrical stimulation offers considerable advantages relating to the physical integrity of the worms (lower mortality rate than that obtained during chemical stimulation), the short duration of stimulation and the ease of recovery of sticky excretions. In addition, the electrical stimulation technique offers the possibility of reusing the worms in other electrical stimulations after a rest period of 3 weeks.

Short-Term Dosage Regimen for Stimulation-Induced Long-Lasting Desynchronization.

In this paper, we computationally generate hypotheses for dose-finding studies in the context of desynchronizing neuromodulation techniques. Abnormally strong neuronal synchronization is a hallmark of several brain disorders. Coordinated Reset (CR) stimulation is a spatio-temporally patterned stimulation technique that specifically aims at disrupting abnormal neuronal synchrony. In networks with spike-timing-dependent plasticity CR stimulation may ultimately cause an anti-kindling, i.e., an unlearning of abnormal synaptic connectivity and neuronal synchrony. This long-lasting desynchronization was theoretically predicted and verified in several pre-clinical and clinical studies. We have shown that CR stimulation with rapidly varying sequences (RVS) robustly induces an anti-kindling at low intensities e.g., if the CR stimulation frequency (i.e., stimulus pattern repetition rate) is in the range of the frequency of the neuronal oscillation. In contrast, CR stimulation with slowly varying sequences (SVS) turned out to induce an anti-kindling more strongly, but less robustly with respect to variations of the CR stimulation frequency. Motivated by clinical constraints and inspired by the spacing principle of learning theory, in this computational study we propose a short-term dosage regimen that enables a robust anti-kindling effect of both RVS and SVS CR stimulation, also for those parameter values where RVS and SVS CR stimulation previously turned out to be ineffective. Intriguingly, for the vast majority of parameter values tested, spaced multishot CR stimulation with demand-controlled variation of stimulation frequency and intensity caused a robust and pronounced anti-kindling. In contrast, spaced CR stimulation with fixed stimulation parameters as well as singleshot CR stimulation of equal integral duration failed to improve the stimulation outcome. In the model network under consideration, our short-term dosage regimen enables to robustly induce long-term desynchronization at comparably short stimulation duration and low integral stimulation duration. Currently, clinical proof of concept is available for deep brain CR stimulation for Parkinson's therapy and acoustic CR stimulation for tinnitus therapy. Promising first in human data is available for vibrotactile CR stimulation for Parkinson's treatment. For the clinical development of these treatments it is mandatory to perform dose-finding studies to reveal optimal stimulation parameters and dosage regimens. Our findings can straightforwardly be tested in human dose-finding studies.

Cerebral haemodynamic response to somatosensory stimulation in near-term fetal sheep.

Cerebral haemodynamic response to neural stimulation has been extensively investigated in animal and clinical studies, in both adult and paediatric populations, but little is known about cerebral haemodynamic functional response in the fetal brain. The present study describes the cerebral haemodynamic response measured by near-infrared spectroscopy to somatosensory stimulation in fetal sheep. The cerebral haemodynamic response in the fetal sheep brain changes from a positive (increase in oxyhaemoglobin (oxyHb)) response pattern to a negative or biphasic response pattern when the duration of somatosensory stimulation is increased, probably due to cerebral vasoconstriction with prolonged stimulations. In contrast to adult studies, we have found that changes in fetal cerebral blood flow and oxyHb are positively increased in response to somatosensory stimulation during hypercapnia. We propose this is related to reduced vascular resistance and recruitment of cerebral vasculature in the fetal brain during hypercapnia. Functional hyperaemia induced by a localised increase in neuronal activity has been suggested to occur in the fetal brain owing to a positive blood oxygen level-dependent (BOLD) signal recorded by functional magnetic resonance imaging following acoustic stimulation. To study the effect of somatosensory input on local cerebral perfusion we used near-infrared spectroscopy (NIRS) in anaesthetised, partially exteriorised fetal sheep where the median nerve was stimulated with trains of pulses (2ms, 3.3Hz) for durations of 1.8, 4.8 and 7.8s. Signal averaging of cerebral NIRS responses to 20 stimulus trains repeated every 60s revealed that a short duration of stimulation (1.8s) increased oxyhaemoglobin in the contralateral cortex consistent with a positive functional response, whereas longer durations of stimulation (4.8, 7.8s) produced more variable oxyhaemoglobin responses including positive, negative and biphasic patterns of change. Mean arterial blood pressure and cerebral perfusion as monitored by laser Doppler flowmetry always showed small, but coincident increases following median nerve stimulation regardless of the type of response detected by the NIRS in the contralateral cortex. Hypercapnia significantly increased the baseline total haemoglobin and deoxyhaemoglobin, and in 7 of 8 fetal sheep positively increased the changes in contralateral total haemoglobin and oxyhaemoglobin in response to the 7.8s stimulus train, compared to the response recorded during normocapnia. These results show that activity-driven changes in cerebral perfusion and oxygen delivery are present in the fetal brain, and persist even during periods of hypercapnia-induced cerebral vasodilatation.

Repetitive Transcranial Magnetic Stimulation (rTMS) for Higher Brain Function Deficits

The management of higher brain dysfunctions such as stroke-induced unilateral spatial neglect (USN) or aphasia is crucial because these dysfunctions have devastating neurological repercussions on the patients' daily life and quality of life. Impairment of the physiological interhemispheric rivalry is often the result of brain insults such as strokes or traumatic injuries, and it may lead to USN or aphasia. Repetitive transcranial magnetic stimulation (rTMS), a non-invasive brain stimulation method, is a promising tool for restoring the pathological imbalance in interhemispheric rivalry by either suppressing the hyperactivity of the unaffected hemisphere or facilitating hypoactivity in the affected hemisphere. The concept of paradoxical functional facilitation (Kapur, 1996) has important clinical implications when coupled with rTMS applications. In addition to conventional rTMS (c-rTMS), other clinically relevant protocols of patterned rTMS (p-rTMS) have been developed: the theta burst stimulation (TBS), the paired associative stimulation (PAS), and the quadripulse stimulation (QPS). TBS is commonly used in the rehabilitation of patients with post-stroke USN and those with non-fluent aphasia because of its prolonged beneficial effects and the short duration of stimulation. TBS may be considered an effective and safe protocol of rTMS. We foresee broader clinical applications of p-rTMS (TBS) and c-rTMS in the treatment of various neurological deficits.

Cell-free DNA in Human Follicular Microenvironment: New Prognostic Biomarker to Predict in vitro Fertilization Outcomes.

Cell-free DNA (cfDNA) fragments, detected in blood and in other biological fluids, are released from apoptotic and/or necrotic cells. CfDNA is currently used as biomarker for the detection of many diseases such as some cancers and gynecological and obstetrics disorders. In this study, we investigated if cfDNA levels in follicular fluid (FF) samples from in vitro fertilization (IVF) patients, could be related to their ovarian reserve status, controlled ovarian stimulation (COS) protocols and IVF outcomes. Therefore, 117 FF samples were collected from women (n = 117) undergoing IVF/Intra-cytoplasmic sperm injection (ICSI) procedure and cfDNA concentration was quantified by ALU-quantitative PCR. We found that cfDNA level was significantly higher in FF samples from patients with ovarian reserve disorders (low functional ovarian reserve or polycystic ovary syndrome) than from patients with normal ovarian reserve (2.7 ± 2.7 ng/μl versus 1.7 ± 2.3 ng/μl, respectively, p = 0.03). Likewise, FF cfDNA levels were significant more elevated in women who received long ovarian stimulation (> 10 days) or high total dose of gonadotropins (≥ 3000 IU/l) than in women who received short stimulation duration (7–10 days) or total dose of gonadotropins < 3000 IU/l (2.4 ± 2.8 ng/μl versus 1.5 ± 1.9 ng/μl, p = 0.008; 2.2 ± 2.3 ng/μl versus 1.5 ± 2.1 ng/μl, p = 0.01, respectively). Finally, FF cfDNA level was an independent and significant predictive factor for pregnancy outcome (adjusted odds ratio = 0.69 [0.5; 0.96], p = 0.03). In multivariate analysis, the Receiving Operator Curve (ROC) analysis showed that the performance of FF cfDNA in predicting clinical pregnancy reached 0.73 [0.66–0.87] with 88% specificity and 60% sensitivity. CfDNA might constitute a promising biomarker of follicular micro-environment quality which could be used to predict IVF prognosis and to enhance female infertility management.

The effect of ultrasonic waves on the phase behavior of a surfactant–brine–oil system

In recent years, most of the mature oilfields in the world have needed enhanced oil recovery (EOR) techniques to maintain their production level. Nevertheless, each EOR technique suffers from a number of limitations. Surfactant flooding, which is a conventional EOR method, can cause emulsification and displacement of the trapped oil in the reservoir. Surfactants are the most expensive components in a microemulsion. Therefore, selecting a proper surfactant formulation which can mobilize oil without considerable surfactant adsorption is very important. The application of ultrasound is one of the unconventional EOR methods. This causes emulsification of oil and water in the reservoir. Integration of surfactant flooding and ultrasound has the potential to decrease surfactant consumption. Therefore, in the case of integrated ultrasound-surfactant flooding, the phase behavior of surfactant–brine–oil, herewith referred to as the SBO system, is crucial and needs to be investigated. In this study, the effect of ultrasonic stimulation duration on the phase behavior of an SBO system was investigated, and changes in solubilization parameters and optimal salinities are discussed. By comparing the results of the phase behavior of SBO under short and long durations of ultrasonic stimulation, it was concluded that a short duration of stimulation (15 min) yields more volume of microemulsion compared to cases using no ultrasound and a longer duration of ultrasound stimulation. Therefore, by using short durations of ultrasonic stimulation in the integrated ultrasound-surfactant flooding process, the interfacial tension remains low and the surfactant consumption decreases.

Event-related electrical stimulation of motor cortices enhances performance in a hand-tapping task

Introduction: Transcranial electrical stimulation (TES) for about 20 min, has resulted in the majority of observed effects in related TES research. While this duration of stimulation has been recognised as safe for human use, some researchers incorporated an alternative method of administration incorpo- rating less than 3 minutes of stimulation, to investigate whether it still produces significant results. It has been shown that short duration stimulation significantly improved verbal memory performance on a word recognition task, in which less than 2 s of anodal stimulation administered per trial was sufficient. Methods: This study sought to investigate whether stimulation locked to Events (‘Event-related’ stimulation) was effective for the improvement of other types of memory, such as procedural motor memory. To test this, response times (RTs) for an implicit sequence- learning hand-tapping task using anodal tDCS over the primary motor cortex (PMC) were compared across Event-related, Constant, and Sham conditions (n ¼ 15 per condition). Participants were asked to tap the buttons on a response box with six buttons as soon as a cue light lit up in the button. Results: Differences in the slopes of RTs over blocks of trials were fitted with an exponential decay function, which revealed a sig- nificant difference in the exponential decay variable, comparing Event-related and Sham conditions (p ¼ 0.042). Discussions: These results show that Event-related stimulation was effective for the improvement of procedural motor memory as compared to Sham. The results of this is another evidence of effectiveness of short duration of stimulation and highlight the importance of synchronisation of stimulation with the ongoing task.

Optimum Stimulation Frequency of High-Frequency Repetitive Transcranial Magnetic Stimulation for Upper-Limb Function in Healthy Subjects

Objective: Because rTMS requires the subject’s head to be immobilized and the subject to maintain the same posture throughout stimulation, stimulation of long duration may induce discomfort. If changing the stimulation parameters can allow the duration of rTMS to be shortened, physical discomfort may decrease. The purpose of this study was to identify the most beneficial stimulation parameters for high-frequency rTMS in terms of the effect on upper-limb function in healthy subjects. Materials and Methods: Forty right-handed healthy volunteers were divided into four groups: three real rTMS groups (5, 10, and 20 Hz rTMS) and one sham group. In the real rTMS groups, 600 impulses were applied at a frequency of 5, 10, or 20 Hz and an intensity of 90% of resting motor threshold. Performance on a peg-board task, tapping task, and grip strength were measured before stimulation, immediately after stimulation, and 20 min after stimulation. Results: All real rTMS groups showed a significant increase in performance on the peg-board task and tapping task after rTMS. There was no significant increase in grip strength in any group. Conclusions: 10-Hz rTMS may improve upper-limb function with a shorter duration of stimulation than rTMS at 5 or 20 Hz. 10-Hz rTMS had the shortest stimulation time, and is recommended as beneficial setting to use with a little discomfort. These results can be used when deciding rTMS stimulation frequency.

Effect of Long Protocol on the Outcome of In Vitro Fertilization-embryo Transfer in Patients with Endometriosis Compared with Prolonged Protocol

Objective To investigate the effect of different down-regulation protocol on the in vitro fertilization-embryo transfer (IVF-ET) outcomes in infertile patients with endometriosis (EMs). Methods A retrospective case control study was performed. Totally 294 infertile patients with EMs were enrolled. And 109 patients (116 cycles) received prolonged protocol as the control, 185 patients (193 cycles) received long protocol as case group, all followed by standard controlled ovarian hyperstimulation (COH). Response to gonadotropins, the fertilization rate, the cleavage rate, the implantation rate, the clinical pregnancy rate and the miscarriage rate were measured and analyzed between the two groups. Results A trend toward better ovarian response was observed in long protocol group. Higher fertilization rate, lower total dose of rFSH, shorter duration of stimulation and more endometrial thickness on the day of hCG injection were observed in long protocol group compared with those of prolonged protocol group, and the difference was significant (P Conclusion Long protocol regimen before IVF-ET in patients with EMs resulted in a trend toward better ovarian response and higher clinical pregnancy rates than prolonged protocol regimen did.

In vitro fertilization stimulation protocol for normal responder patients

The aim of this prospective observational study is to determine the different outcomes of IVF/ICSI treatments after using antagonists or agonists of gonadotrophin-releasing hormone (GnRH) for controlled ovarian hyperstimulation (COH) in normal responder patients. Two hundred forty-seven patients undergoing IVF treatment at the Centre of Reproductive Medicine, Rome (CERMER), from January 2005 to December 2008, were included in the study. Patients were stimulated either with a standard long protocol with GnRH agonists (n = 156) or with GnRH antagonists (n = 91). The use of GnRH antagonists resulted in a significant reduction in the duration of the stimulation (Agonist Group 14.10 ± 2.25 vs Antagonist Group 11.34 ± 2.11; p < 0.001) and in the amount of gonadotrophin (IU of r-FSH) needed (Agonist Group 1878 ± 1109 vs Antagonist Group 1331 ± 1049; p = 0.0014). Moreover a lower number of cycles were cancelled with the antagonist protocol (4.39 vs 6.41%). The GnRH antagonist protocol, when compared to the GnRH agonist one, is associated with a similar clinical pregnancy rate, similar implantation rate, significantly lower gonadotrophin requirement and shorter duration of stimulation. For this reason, GnRH antagonists might be a good treatment even for normal responder patients undergoing IVF.

Long Lasting Modulation of Cortical Oscillations after Continuous Theta Burst Transcranial Magnetic Stimulation

Transcranial magnetic theta burst stimulation (TBS) differs from other high-frequency rTMS protocols because it induces plastic changes up to an hour despite lower stimulus intensity and shorter duration of stimulation. However, the effects of TBS on neuronal oscillations remain unclear. In this study, we used electroencephalography (EEG) to investigate changes of neuronal oscillations after continuous TBS (cTBS), the protocol that emulates long-term depression (LTD) form of synaptic plasticity. We randomly divided 26 healthy humans into two groups receiving either Active or Sham cTBS as control over the left primary motor cortex (M1). Post-cTBS aftereffects were assessed with behavioural measurements at rest using motor evoked potentials (MEPs) and at active state during the execution of a choice reaction time (RT) task in combination with continuous electrophysiological recordings. The cTBS-induced EEG oscillations were assessed using event-related power (ERPow), which reflected regional oscillatory activity of neural assemblies of θ (4–7.5 Hz), low α (8–9.5 Hz), µ (10–12.5 Hz), low β (13–19.5 Hz), and high β (20–30 Hz) brain rhythms. Results revealed 20-min suppression of MEPs and at least 30-min increase of ERPow modulation, suggesting that besides MEPs, EEG has the potential to provide an accurate cortical readout to assess cortical excitability and to investigate the interference of cortical oscillations in the human brain post-cTBS. We also observed a predominant modulation of β frequency band, supporting the hypothesis that cTBS acts more on cortical level. Theta oscillations were also modulated during rest implying the involvement of independent cortical theta generators over the motor network post cTBS. This work provided more insights into the underlying mechanisms of cTBS, providing a possible link between synchronised neural oscillations and LTD in humans.