Data from multiple sclerosis (MS) and the MS rodent model, experimental autoimmune encephalomyelitis (EAE), highlighted an inflammation-dependent synaptopathy at the basis of the neurodegenerative damage causing irreversible disability in these disorders. This synaptopathy is characterized by an imbalance between glutamatergic and GABAergic transmission and has been proposed to be a potential therapeutic target. Siponimod (BAF312), a selective sphingosine 1-phosphate1,5 receptor modulator, is currently under investigation in a clinical trial in secondary progressive MS patients. We investigated whether siponimod, in addition to its peripheral immune modulation, may exert direct neuroprotective effects in the central nervous system (CNS) of mice with chronic progressive EAE. Minipumps allowing continuous intracerebroventricular (icv) infusion of siponimod for 4 weeks were implanted into C57BL/6 mice subjected to MOG35-55-induced EAE. Electrophysiology, immunohistochemistry, western blot, qPCR experiments, and peripheral lymphocyte counts were performed. In addition, the effect of siponimod on activated microglia was assessed in vitro to confirm the direct effect of the drug on CNS-resident immune cells. Siponimod administration (0.45 μg/day) induced a significant beneficial effect on EAE clinical scores with minimal effect on peripheral lymphocyte counts. Siponimod rescued defective GABAergic transmission in the striatum of EAE, without correcting the EAE-induced alterations of glutamatergic transmission. We observed a significant attenuation of astrogliosis and microgliosis together with reduced lymphocyte infiltration in the striatum of EAE mice treated with siponimod. Interestingly, siponimod reduced the release of IL-6 and RANTES from activated microglial cells in vitro, which might explain the reduced lymphocyte infiltration. Furthermore, the loss of parvalbumin-positive (PV+) GABAergic interneurons typical of EAE brains was rescued by siponimod treatment, providing a plausible explanation of the selective effects of this drug on inhibitory synaptic transmission. Altogether, our results show that siponimod has neuroprotective effects in the CNS of EAE mice, which are likely independent of its peripheral immune effect, suggesting that this drug could be effective in limiting neurodegenerative pathological processes in MS.

Purpose: We evaluated leukemia-associated immunophenotypes (LAIP) and their correlation with fms-like tyrosine kinase 3 (FLT3) and nucleophosmin (NPM1) gene mutational status in order to contribute a better identification of patients at highest risk of relapse in acute myeloid leukemia (AML).Experimental Design: Bone marrow samples from 132 patients with AML were analyzed by nine-color multiparametric flow cytometry. We confirmed the presence of the mutation in diagnostic samples and in sorted cells by conventional RT-PCR and by patient-specific RQ-PCR.Results: Within the CD34+ cell fraction, we identified a discrete population expressing high levels of the IL3 receptor α-chain (CD123) and MIC-2 (CD99) in combination with the IL2 receptor α-chain (CD25). The presence of this population positively correlated with the internal tandem duplications (ITD) mutation in the FLT3 gene (r = 0.71). Receiver operating characteristics showed that, within the CD34+ cell fraction a percentage of CD123/CD99/CD25+ cells ≥11.7% predicted FLT3–ITD mutations with a specificity and sensitivity of >90%. CD34/CD123/CD99/CD25+ clones were also detectable at presentation in 3 patients with FLT3 wild-type/NPM1+ AML who relapsed with FLT3-ITD/NPM1+ AML. Quantitative real-time PCR designed at relapse for each FLT3-ITD in these three cases confirmed the presence of low copy numbers of the mutation in diagnostic samples.Conclusions: Our results suggest that the CD34/CD25/CD123/CD99+ LAIP is strictly associated with FLT3-ITD–positive cells. Clin Cancer Res; 21(17); 3977–85. ©2015 AACR.

Forkhead box P3 (FoxP3)+ regulatory T cells (Treg) are powerful mediators of immune regulation and immune homeostasis. In humans, Tregs are a heterogeneous population expressing surface markers which define phenotypically and functionally distinct subsets. Moreover, it is now clear that intracellular staining for FoxP3 does not unequivocally identify “true” suppressor cells, since several FoxP3 isoforms exist, and different reagents for FoxP3 detection are available. Here, we propose a strategy to identify potentially functional and suppressive Treg cells in an autoimmune disease like multiple sclerosis, and we suggest that in patients affected by this disease these cells are both reduced in number and functionally exhausted.

Epidural blockade is a method often used in clinical practice to obtain pain relief during labor. Basically, the epidural puncture blocks the nerve impulses from the lower spinal segments with a consequent decrease of sensation in the lower half of the body. In this arena, the correct localization of the epidural space is crucial to perform the procedure. The loss of resistance (LOR) technique is the most popular approach to localize the epidural space. The LOR is caused by the fact that the ligamentum flavum is much harder than the epidural space, therefore when the needle tip pierces this ligamentum, there is an abrupt decrease of pressure (known as LOR). Although, there are several commercially available tools to support the clinicians in this task, the large part of epidural punctures is performed without any support. Our group of research has developed a novel and noninvasive tool to detect the LOR during epidural puncture. The aim of this study is to compare the output collected by the proposed system (i.e., the Epidural Space Management Tool, ESMT) with the output provided by a commercial system (i.e., CompuFlo®, Milestone Scientific, Inc.). Experiments were performed by an expert anesthesiologist (M.C.) on a simple simulator that allows mimicking the real scenario. Results showed a good agreement between the trends of the output of the two mentioned systems. Finally, the proposed system (ESMT), showed promising results in terms of LOR detection during all the five procedures performed on the simulator.

Background and Aims Difficult airway may represent a challenge in anesthesiologists’ clinical practice. In some cases regional anesthesia might be an option. Given a preformulated strategy for intubation, the right decision mainly depends on how confident the anesthesiologist is with the appropriate regional technique [1, 2]. We describe the case of an impossible to intubate patient, underwent surgical debridement of a neck ulcer under regional anesthesia. Methods Male 57 y.o. History of nasopharyngeal carcinoma, treated with radiotherapy. Post-actinic osteonecrosis of the jaw; scar tissue around muscles and vessels of the left side of the neck with consequent lock-jaw and cervical rigidity. Post-actinic infected lesion of the skin in the lateral, posterior and deltoid regions of the left side of the neck, scheduled for surgical debridement and vac-therapy. Awake tracheostomy was established as anesthetic management. The patient refused and asked for alternatives.Regional anesthesia was considered. Superficial or intermediate cervical plexus block were not feasible because of the scar tissue. Thus, to cover the area of debridement, a combination of a left ESP block at T2 level and a left deep cervical plexus block at C4 were performed. Awake, trans nasal fiberoptic intubation, was considered as preformulated strategy in case of emergency [3]. Results The surgical procedure was carried out uneventfully with the patient awake and cooperative. Conclusions Performing regional anesthesia in the difficult airways patients implies major knowledge of the relevant anatomy and techniques. Major skills in airways management are also required. Given these two conditions, it is not always foolish not to secure the airway.

Background and Objectives: The aim of this study was to evaluate the impact of medications on oxidative stress, inflammatory biomarkers and semen characteristics in males with idiopathic infertility. Materials and Methods: In this observational case-control clinical study, 50 men with idiopathic infertility were enrolled, of whom 38 (the study group) were on pharmacological treatment and 12 made up the control group. The study group was clustered according to the medications (Group A: anti-hypertensive, n = 10; Group B: thyroxine, n = 6; Group C: non-steroidal anti-inflammatory drugs, n = 13; Group D: miscellaneous, n = 6; Group E: lipid-lowering drugs, n = 4). Semen analyses were performed according to WHO 2010 guidelines. Interleukins (IL)-10, IL-1 beta, IL-4, IL-6, Tumor Necrosis Factor- alpha (TNF-alpha) and IL-1 alpha were determined using a solid-phase sandwich immunoassay. The diacron reactive oxygen metabolites, d-ROMs test, was performed by means of a colorimetric determination of reactive oxygen metabolites and measured with a spectrophotometer. Beta-2-microglobulin and cystatin-C were measured with an immunoturbidimetric analyzer. Results: No differences between the study and control groups for age and macroscopic and microscopic semen characteristics were found, nor were any differences found after clustering according to the drug categories. IL-1 alpha and IL-10 were significantly lower in the study group compared with the control group; IL-10 was significantly lower in groups A, B, C and D compared with the control group. Furthermore, a direct correlation between IL-1 alpha, IL-10 and TNF-alpha and leukocytes was found. Conclusions: Despite the sample size limitations, the data suggest a correlation between drug use and activation of the inflammatory response. This could clarify the pathogenic mechanism of action for several pharmacological classes on male infertility.