Idiopathic Generalized Epilepsy (IGE) management has become increasingly challenging due to the restricted use of Valproate (VPA) in females. The aim of the study was to assess possible differences ...in terms of seizure outcome between men and women suffering from IGE.
A cohort of IGE patients (age range: 13–50 years) followed from 1980 to 2018 were included. Their medical history was retrospectively reviewed to investigate possible factors influencing seizure outcome. Seizure Remission (SR) was defined as the absence of any seizure type over 18 months prior to the last medical observation. The primary outcome was to evaluate sex differences in terms of SR at last observation. A multivariable logistic regression model was elaborated using SR as dependent variable.
Three-hundred and sixty patients were included, 204 (56.7%) of whom were women. The median age at the end of follow-up was 30. At last medical observation, fewer women were receiving VPA compared with men (females 39.7% vs males 79.5%, p < .001). Overall SR was 70.6%. SR was significantly different according to sex (females 62.3% vs males 81.4%, p < .001). Multivariable logistic regression model showed that female sex (Odds Ratios OR = 0.52, 95% Confidence Interval CI = 0.29–0.94; p = .03), VPA treatment at last observation (OR = 0.44, 95% CI = 0.25–0.76; p = .003) and epilepsy syndrome (p < .001) were the factors independently associated with SR.
Recent modifications in VPA prescribing patterns may have determined a worse seizure control among IGE female patients. Comparative clinical trials assessing the best therapeutic options for women with childbearing potential are urgently needed.
•Therapeutic management is challenging in IGE female patients of childbearing age.•Given the limitations in VPA use, we investigated sex differences in seizure outcome.•We found a significant difference in VPA use between females and males.•VPA dosage and blood level were also found to differ according to gender.•Male sex and VPA use were independently associated with a better seizure outcome.
Abstract
Background
Sjögren’s syndrome is a systemic autoimmune disease characterized by immune cells predominantly infiltrating the exocrine glands and frequently forming ectopic lymphoid ...structures. These structures drive a local functional immune response culminating in autoantibody production and tissue damage, associated with severe dryness of mucosal surfaces and salivary gland hypofunction. Cenerimod, a potent, selective and orally active sphingosine-1-phosphate receptor 1 modulator, inhibits the egress of lymphocytes into the circulation. Based on the mechanism of action of cenerimod, its efficacy was evaluated in two mouse models of Sjögren’s syndrome.
Methods
Cenerimod was administered in two established models of Sjögren’s syndrome; firstly, in an inducible acute viral sialadenitis model in C57BL/6 mice, and, secondly, in the spontaneous chronic sialadenitis MRL/lpr mouse model. The effects of cenerimod treatment were then evaluated by flow cytometry, immunohistochemistry, histopathology and immunoassays. Comparisons between groups were made using a Mann-Whitney test.
Results
In the viral sialadenitis model, cenerimod treatment reduced salivary gland immune infiltrates, leading to the disaggregation of ectopic lymphoid structures, reduced salivary gland inflammation and preserved organ function. In the MRL/lpr mouse model, cenerimod treatment decreased salivary gland inflammation and reduced T cells and proliferating plasma cells within salivary gland ectopic lymphoid structures, resulting in diminished disease-relevant autoantibodies within the salivary glands.
Conclusions
Taken together, these results suggest that cenerimod can reduce the overall autoimmune response and improve clinical parameters in the salivary glands in models of Sjögren’s syndrome and consequently may reduce histological and clinical parameters associated with the disease in patients.
Abstract
Pigs are becoming an important pre-clinical animal species for translational ophthalmology, due to similarities with humans in anatomical and physiological patterns. Different models of eye ...disorders have been proposed, and they are good candidates to assess biocompatibility/functionality of retinal prostheses. Electroretinography is a common tool allowing to gain information on retinal function, with several types of electroretinogram (ERG) been implemented including full field (ff-ERG), multifocal (mf-ERG) and pattern (p-ERG). p-ERG represents a valuable tool to monitor Retinal Ganglion Cells (RGCs) activity and can be used to calculate p-ERG spatial acuity. Unfortunately, scarce methodological data are available regarding recording/interpretation of p-ERG and retinal acuity in biomedical pigs yet enhancing knowledge regarding pig vision physiology will allow for more refined and responsible use of such species. Aim of this study was to record p-ERG in juvenile pigs to functionally assess visual acuity. Six female hybrid pigs underwent two p-ERG recording sessions at 16 and 19 weeks of age. Photopic ff-ERG were also recorded; optical coherence tomography (OCT) and histology were used to confirm retinal integrity. ff-ERG signals were repeatable within/across sessions. All p-ERG traces consistently displayed characterizing peaks, and the progressive decrease of amplitude in response to the increment of spatial frequency revealed the reliability of the method. Mean p-ERG spatial acuities were 5.7 ± 0.14 (16 weeks) and 6.2 ± 0.15 cpd (19 weeks). Overall, the p-ERG recordings described in the present work seem reliable and repeatable, and may represent an important tool when it comes to vision assessment in pigs.
Essential oils possess a variety of biological properties (i.e., antioxidant, antibacterial, and cytotoxic) that could possibly be applied in reproductive medicine, but their effects on spermatozoa ...are still partially unknown. The aim of the study was to describe the effects of
(L.) Cav. and
(L.) essential oils on the main morpho-functional parameters of swine spermatozoa. Essential oils were preliminary characterized by gas chromatography and added with emulsifiers to facilitate diffusion. Experimental samples were prepared by suspending a fixed number of spermatozoa in 5 mL of medium with 10 different concentrations of essential oil (0.2-2 mg/mL, at intervals of 0.2). After 3 h of incubation, samples were analyzed for pH, viability, objective motility, and acrosome status. Results showed that the effects of the essential oils are concentration-dependent and that
is well tolerated up to 0.6 mg/mL.
impaired the spermatozoa starting from the lowest concentration, with complete spermicidal effect from 0.4 mg/mL. The patterns of damage, confirmed by SEM, were different and quite distinct. As expected, spermatozoa proved to be sensitive to external stimuli and capable of showing different functional patterns, providing interesting insights to the action/toxicity mechanisms. The results of the present work represent the first step towards the systematic characterization of the effects of these compounds on spermatozoa. This kind of studies are necessary to strengthen the idea of future applications of essential oils in the reproductive field due to their antioxidant, antibacterial, or spermicidal properties.
Primary Sjögren's syndrome (pSS) is a complex autoimmune disease characterised by local exocrine glandular involvement and systemic multiorgan manifestations. In this review we will discuss the value ...of the histological examination of the salivary glands in the classification criteria, and more recently as prognostic tool for patient stratification and monitoring. The limitations of the current tools used to assess salivary gland pathology in pSS will also be reviewed in relation to using salivary gland biopsy analysis as an outcome measure in clinical trials.
Primary Sjögren's Syndrome (pSS) affects exocrine glands such as those producing the tear film, leading to dry and painful eyes, but is also associated with fatigue. The experience of fatigue in pSS, ...and its relationship with sicca symptoms, is poorly understood.
Twenty people diagnosed with pSS were recruited to participate in a semi-structured qualitative interview about their symptoms experience. Interviews were audio-recorded, transcribed verbatim and analysed using thematic analysis.
People with pSS described physical tiredness, mental fatigue and ocular fatigue. Mental fatigue was characterised by difficulties in attention, particularly, the ability to follow conversations and short-term memory problems. Participants linked their experience of fatigue to feeling of depression, frustration, irritation and anxiety, and therefore, fatigue was suggested to have had a large impact on their psychological well-being. People with pSS also described a range of ocular symptoms including pain, dryness, and itching, which were compounded by fatigue. For some, eye fatigue was pervasive, and daily activities involving the eyes such as reading, using the computer and driving were impaired. In some cases, the level of ocular discomfort was so severe it prevented sleep, which in turn impacted on general fatigue levels.
People with pSS experience fatigue in a range of ways; physical, mental and ocular fatigue were described. Fatigue was suggested to exacerbate other ocular symptoms, posed serious physical limitations and caused psychological distress. Further research into the nature of fatigue and ocular symptoms in pSS is required.
The C-type lectin receptor CLEC-2 signals through a pathway that is critically dependent on the tyrosine kinase Syk. We show that homozygous loss of either protein results in defects in brain ...vascular and lymphatic development, lung inflation, and perinatal lethality. Furthermore, we find that conditional deletion of Syk in the hematopoietic lineage, or conditional deletion of CLEC-2 or Syk in the megakaryocyte/platelet lineage, also causes defects in brain vascular and lymphatic development, although the mice are viable. In contrast, conditional deletion of Syk in other hematopoietic lineages had no effect on viability or brain vasculature and lymphatic development. We show that platelets, but not platelet releasate, modulate the migration and intercellular adhesion of lymphatic endothelial cells through a pathway that depends on CLEC-2 and Syk. These studies found that megakaryocyte/platelet expression of CLEC-2 and Syk is required for normal brain vasculature and lymphatic development and that platelet CLEC-2 and Syk directly modulate lymphatic endothelial cell behavior in vitro.
The importance of CLEC-2, a natural ligand/receptor for Gp38/Podoplanin, in the formation of the lymphatic vasculature has recently been demonstrated. As the development and maintenance of lymph ...nodes (LNs) is dependent on the formation of the lymphatic vasculature and the differentiation of Gp38/Podoplanin+ stromal cells, we investigated the role of CLEC-2 in lymphoneogenesis and LN homeostasis. Using constitutive Clec1b−/− mice, we showed that while CLEC-2 was not necessary for initiation of the LN anlage, it was required at late stages of development. Constitutive deletion of CLEC-2 induced a profound defect in lymphatic endothelial cell proliferation, resulting in lack of LNs at birth. In contrast, conditional deletion of CLEC-2 in the megakaryocyte/platelet lineage in Clec1bfl/flPF4-Cre mice led to the development of blood-filled LNs and fibrosis, in absence of a proliferative defect of the lymphatic endothelial compartment. This phenotype was also observed in chimeric mice reconstituted with Clec1bfl/flPF4-Cre bone marrow, indicating that CLEC-2 expression in platelets was required for LN integrity. We demonstrated that LNs of Clec1bfl/flPF4-Cre mice are able to sustain primary immune responses but show a defect in immune cell recirculation after repeated immunizations, thus suggesting CLEC-2 as target in chronic immune response.
•CLEC-2 is necessary for lymphatic cell proliferation and lymph node anlage persistence after birth.•Lack of CLEC-2 expression on megakaryocytes and platelets compromises lymph node integrity in adult life.
BACKGROUNDPrimary Sjögren's syndrome (pSS) is characterized by B cell hyperactivity and elevated B-lymphocyte stimulator (BLyS). Anti-BLyS treatment (e.g., belimumab) increases peripheral memory B ...cells; decreases naive, activated, and plasma B cell subsets; and increases stringency on B cell selection during reconstitution. Anti-CD20 therapeutics (e.g., rituximab) bind and deplete CD20-expressing B cells in circulation but are less effective in depleting tissue-resident CD20+ B cells. Combined, these 2 mechanisms may achieve synergistic effects.METHODSThis 68-week, phase II, double-blind study (GSK study 201842) randomized 86 adult patients with active pSS to 1 of 4 arms: placebo, s.c. belimumab, i.v. rituximab, or sequential belimumab + rituximab.RESULTSOverall, 60 patients completed treatment and follow-up until week 68. The incidence of adverse events (AEs) and drug-related AEs was similar across groups. Infections/infestations were the most common AEs, and no serious infections of special interest occurred. Near-complete depletion of minor salivary gland CD20+ B cells and a greater and more sustained depletion of peripheral CD19+ B cells were observed with belimumab + rituximab versus monotherapies. With belimumab + rituximab, reconstitution of peripheral B cells occurred, but it was delayed compared with rituximab. At week 68, mean (± standard error) total EULAR Sjögren's syndrome disease activity index scores decreased from 11.0 (1.17) at baseline to 5.0 (1.27) for belimumab + rituximab and 10.4 (1.36) to 8.6 (1.57) for placebo.CONCLUSIONThe safety profile of belimumab + rituximab in pSS was consistent with the monotherapies. Belimumab + rituximab induced enhanced salivary gland B cell depletion relative to the monotherapies, potentially leading to improved clinical outcomes.TRIAL REGISTRATIONClinicalTrials.gov NCT02631538.FUNDINGFunding was provided by GSK.
BackgroundThe CD47/SIRPα axis mediates a ‘don’t eat me’ signal exploited by tumor cells to escape macrophage-mediated immune surveillance. Anti-CD47 therapies have shown promising clinical results in ...solid and hematological malignancies; however, efficacy is hindered by systemic toxicity. We hypothesized that local delivery of a therapeutic, able to interfere with the CD47/SIRPα axis within an oncolytic viral chassis, would induce high payload expression paired with oncolytic activity and low systemic exposure, ultimately resulting in improved tumor control.MethodsAlpha-201-macro1 is a first-in-class, replication-defective oncolytic virus encoding a protein payload able to interfere with the CD47/SIRPα axis. Disruption of the interaction between CD47 and SIRPα was confirmed in vitro using a plate-based CD47 displacement assay. Bioactivity was tested in an ex vivo phagocytosis assay using Raji cells and differentiated M1 macrophages treated with conditioned medium obtained from tumor cells infected with Alpha-201-macro1 or control vectors. Phagocytosis was measured by flow cytometry. Anti-tumor efficacy of Alpha-201-macro1, delivered intratumorally at 3x107 PFU, was assessed in A549 tumor-bearing BALB/c-Nude mice (N=8 per group) and compared to anti-CD47 antibody therapy (clone: B6H12; intraperitoneal, 10 mg/kg). Treatments were administered on day 1, 4, 7, 10, and 13 after randomization. All values are reported as mean±SEM.ResultsConditioned media from Alpha-201-macro1 infected cells selectively disrupted binding of SIRPα to CD47 in a payload- (and not vector-) dependent manner. Additionally, conditioned medium from Alpha-201-macro1 infected cells resulted in a dose-dependent increase in macrophage-mediated target cell phagocytosis (3 PFU/cell: 4.16±0.17%, 10 PFU/cell: 17.8±2.55%) that was greater than the effect of the anti-CD47 antibody (6.86±1.68%, p=0.017 compared to 10 PFU/cell Alpha-201-macro1). Pretreatment with conditioned medium from Alpha-201 vectors encoding irrelevant payloads did not significantly affect macrophage phagocytosis, demonstrating the specificity of the effect of the therapeutic payload. In vivo, treatment with Alpha-201-macro1 resulted in tumor growth inhibition compared to the vehicle control group (217.1±17.4 mm3 vs. 312.7±30.5 mm3, p=0.023). This effect was dependent on payload expression, as a control vector did not show statistically significant tumor activity. There was a trend towards greater efficacy with local Alpha-201-macro1 delivery compared to systemic anti-CD47 antibody (217.1±17.4 mm3 vs. 248.7±27.9 mm3, p=0.38).ConclusionsAlpha-201-macro1 enhances macrophage phagocytosis ex vivo and exerts anti-tumor efficacy in vivo, effects which exceed those of a systemically administered anti-CD47 antibody. Further in vivo studies of Alpha-201-macro1 and modified, multi-payload versions of this vector, in combination with immune checkpoint inhibitors, are ongoing.Ethics ApprovalAll procedures involving the care and use of animals in this study were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC).
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