Lamotrigine treatment of a patient affected by epilepsy and anxiety disorder

Epilepsy often occurs in comorbidity with mental diseases and disorders. Early detection and/or treatment of such disorders in patients affected by epilepsy, as well as their socialisation are crucially important since epileptic patients tend to suffer more due to lack of social support than to frequent epileptic seizures. Prevalence of psychiatric disorders is higher in patients with epilepsy than in general population, the most frequent being: anxiety, depression, panic attacks, behavioural disorders as well as psychotic states with paranoid elements. The efficacy of AE treatment of patients affected by epilepsy and mood disorders has also directed clinicians to investigate possible AE benefits in treating other mental disorders such as anxiety states, depression and bipolar disorder. The examined case displays complex partial epilepsy and comorbid mental disorder. The use of lamotrigine, a fourth-generation antiepileptic, which is also a mood stabilizer, has assured a favourable remission of symptoms related to both epilepsy and mood disorders. Side-effects caused by lamotrigine were only temporary and dose reduction was sufficient to eliminate their symptoms.     

ID1 and ID3 regulate the self-renewal capacity of human colon cancer-initiating cells through p21

There is increasing evidence that some cancers are hierarchically organized, sustained by a relatively rare population of cancer-initiating cells (C-ICs). Although the capacity to initiate tumors upon serial transplantation is a hallmark of all C-ICs, little is known about the genes that control this process. Here, we establish that ID1 and ID3 function together to govern colon cancer-initiating cell (CC-IC) self-renewal through cell-cycle restriction driven by the cell-cycle inhibitor p21. Regulation of p21 by ID1 and ID3 is a central mechanism preventing the accumulation of excess DNA damage and subsequent functional exhaustion of CC-ICs. Additionally, silencing of ID1 and ID3 increases sensitivity of CC-ICs to the chemotherapeutic agent oxaliplatin, linking tumor initiation function with chemotherapy resistance.     

Telemonitoring of Elderly with Hypertension and Type 2 Diabetes at the Primary Care Level: Protocol for a Multicentric Randomized Controlled Pilot Study

IntroductionArterial hypertension (AH) and type 2 diabetes (T2D) represent a significant burden for the public health system, with an exceptionally high prevalence in patients aged ≥65 years. This study aims to test the acceptability, clinical effectiveness, and cost-effectiveness of telemonitoring in elderly patients with AH and T2D at the primary care level.MethodsA m ulti-centre, prospective, randomized, controlled t rial w ill be conducted. Patients a ged ≥ 65 y ears with AH and T2D will be randomized in a 1:1 proportion to a mHealth intervention or standard care group. Patients in the intervention group will measure their blood pressure (BP) twice weekly and blood glucose (BG) once monthly. The readings will be synchronously transmitted via a mobile application to the telemonitoring platform, where they will be reviewed by a general practitioner who will indicate changes in measurement regimen or carry out a teleconsultation. The primary endpoint will be a change in systolic BP (SBP) and glycated haemoglobin (HbA1c) relative to standard care up to 12 months after inclusion. Secondary endpoints will be a change in other observed clinical variables, quality-of-life indexes, and costs.Expected resultsTelemonitoring will be an acceptable method of care associated with significant reductions in SBP and HbA1c levels and an increase in quality-of-life indexes in the intervention group. However, the cost-effectiveness threshold (incremental cost-effectiveness ratio below €25,000/quality-adjusted life year) might not be reached.ConclusionThis study will provide new evidence for scaling up telemonitoring network at the primary care level and modifying telemonitoring protocols to achieve the best clinical and cost-effective outcomes.     

MK2 degradation as a sensor of signal intensity that controls stress-induced cell fate

Cell survival in response to stress is determined by the coordination of various signaling pathways. The kinase p38α is activated by many stresses, but the intensity and duration of the signal depends on the stimuli. How different p38α-activation dynamics may impact cell life/death decisions is unclear. Here, we show that the p38α-signaling output in response to stress is modulated by the expression levels of the downstream kinase MK2. We demonstrate that p38α forms a complex with MK2 in nonstimulated mammalian cells. Upon pathway activation, p38α phosphorylates MK2, the complex dissociates, and MK2 is degraded. Interestingly, transient p38α activation allows MK2 reexpression, reassembly of the p38α–MK2 complex, and cell survival. In contrast, sustained p38α activation induced by severe stress interferes with p38α–MK2 interaction, resulting in irreversible MK2 loss and cell death. MK2 degradation is mediated by the E3 ubiquitin ligase MDM2, and we identify four lysine residues in MK2 that are directly ubiquitinated by MDM2. Expression of an MK2 mutant that cannot be ubiquitinated by MDM2 enhances the survival of stressed cells. Our results indicate that MK2 reexpression and binding to p38α is critical for cell viability in response to stress and illustrate how particular p38α-activation patterns induced by different signals shape the stress-induced cell fate.

Cells can respond to stress in a variety of ways through the activation of particular signaling pathways. The initial response is usually aimed at protecting the cell against the insult to facilitate damage recovery and cell survival. However, if the harmful stimulus persists or is not properly resolved, a death program is usually activated that eventually eliminates the damaged cells.A signaling pathway that is frequently associated with the stress response involves activation of the mitogen-activated protein kinase (MAPK) family member p38α. Upon activation generally achieved by dedicated MAP2Ks, p38α can phosphorylate a variety of substrates in the nucleus and cytoplasm (1, 2). In particular, the activation of p38α is often linked to the activation of MAPK-activated protein kinase 2 (MAPKAPK2 or MK2), and both kinases regulate several stress responses that impinge on cell survival or cell death (3–6). In addition, the p38α–MK2 pathway plays an important role in the regulation of the immune response, and it can also control the proliferation or differentiation of some cell types, which may contribute to physiological responses not necessarily related to stress (7–9). How this pathway can mediate so many different cellular processes is still largely an open question. In many cases, the cellular response mediated by the p38α–MK2 pathway can be determined by the cell type and the stimuli and often engages particular downstream targets. However, other factors, such as signaling dynamics, may also play a role. It has been reported that some MAPKs can perform different functions depending on the amplitude, duration, and frequency of pathway activation (10), but it is still not known whether the p38α-activation dynamics could modulate the cell responses associated with different stimuli.The formation of protein–protein complexes is important for many biological processes, including signal transduction (11, 12), and changes in their interaction dynamics are crucial for proper sensing of the environmental changes (13). Structural analysis revealed that purified recombinant p38α and MK2 proteins can interact through the p38α docking groove and the MK2 docking motif (14), and further studies addressed the requirements for the formation of the p38α–MK2 complex in vitro (15, 16). Moreover, NMR and X-ray crystallography analysis suggested that p38α-MK2 can form different heterodimers depending on the activation state of p38α (17). However, little is known about how this interaction could be regulated in vivo and whether it modulates the pathway functions.Given the importance of p38α and MK2 in many cellular processes, the activity of both kinases should be tightly controlled. The down-regulation of p38α activity is known to involve several phosphatases and negative feedback loops (18–22). However, it is not clear how MK2 activity is normally down-regulated.In this study, we present evidence that the extent of p38α activation regulates MK2 protein levels, which, in turn, have a key role in the pathway output. We show that endogenous p38α and MK2 form a complex in mammalian cells and that, upon p38α activation, the complex dissociates and MK2 is degraded. In response to mild stress or to physiological stimuli, the pathway is transiently activated, allowing reformation of the p38α–MK2 complex and concomitant cell survival. However, cells treated with severe stress show sustained p38α activation, which interferes with p38α–MK2 interaction, leading to the irreversible loss of MK2 and cell death. Therefore, our results illustrate an additional mechanism of p38α–MK2 pathway regulation, which might help to predict the cell fate in response to stress.     

Thrombin stimulation of p38 MAP kinase in human platelets is mediated by ADP and thromboxane A2 and inhibited by cGMP/cGMP-dependent protein kinase

p38 MAP kinase in human platelets is activated by platelet agonists including thrombin, thromboxane A2 (TxA2), ADP, and others. However, both upstream mechanisms of p38 MAP kinase activation, and their downstream sequelae, are presently controversial and essentially unclear. Certain studies report sequential activation of cGMP-dependent protein kinase (PKG) and p38/ERK pathways by platelet agonists, leading to integrin activation and secretion, whereas others establish an essential role of Src/ERK-mediated TxA2 generation for fibrinogen receptor activation in human platelets. Here, we show that ADP secreted from platelet-dense granules, and subsequent activation of P2Y12 receptors, as well as TxA2 release are important upstream mediators of p38 MAP kinase activation by thrombin. However, p38 MAP kinase activation did not significantly contribute to calcium mobilization, P-selectin expression, alphaIIbbeta3 integrin activation, and aggregation of human platelets in response to thrombin. Finally, PKG activation did not stimulate, but rather inhibited, p38 MAP kinase in human platelets.     

Synthesis, X-ray crystal structure study, and cytostatic and antiviral evaluation of the novel cycloalkyl-N-aryl-hydroxamic acids

In vitro evaluation of the novel cycloalkyl-N-(4-chlorophenyl)-hydroxamic acids (2a-g) demonstrated that 2b,d,e exhibited rather marked inhibitory activity (IC50 = 7-10 microM) against pancreatic carcinoma, 2b-d against colon carcinoma, 2d against laryngeal carcinoma, and 2b,d against breast carcinoma. 2e showed the most pronounced anti-cytomegalovirus activity (EC50 = 1.5 and 0.8 microg mL(-1)) only at > or = 5-fold lower than the cytotoxic concentration. 2d and 2f showed modest, albeit selective, activity against cytomegalovirus (2d, EC50 = 7.3-8.9 microg mL(-1), selectivity index 7-10; 2f, EC50 = 7-13 microg mL(-1), selectivity index 10).     

Potent inhibition of human platelets by cGMP analogs independent of cGMP-dependent protein kinase

Platelets play a key role in hemostasis through their ability to rapidly adhere to activated or injured endothelium, subendothelial matrix proteins, and other activated platelets. A strong equilibrium between activating and inhibiting processes is essential for normal platelet and vascular function, impairment of this equilibrium being associated with either thrombophilic or bleeding disorders. Both cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) have been established as crucial and synergistic intracellular messengers that mediate the effects of platelet inhibitors such as nitric oxide (NO) and prostacyclin (PG-I2). However, it was recently suggested that a rapid cGMP/cGMP-dependent protein kinase (cGK)-mediated extracellular signal-related kinase (ERK) phosphorylation promotes platelet activation. This hypothesis was examined here by evaluating established and proposed cGK activators/inhibitors with respect to their capacity to promote either platelet activation or inhibition. In particular, the regulatory role of cGK for ERK phosphorylation and thrombin-, thromboxane-, and VWF-induced platelet activation was investigated. The data obtained do not support the concept that cGK-mediated ERK phosphorylation promotes platelet activation but confirm the inhibitory role of cGK in platelet function. One explanation for these discrepancies is the novel finding that extracellular cGMP analogs potently and rapidly inhibit thrombin-, thromboxane-, and VWF-induced human platelet signaling and activation by a cGK-independent mechanism.     

Heart rate variability and nonlinear dynamic analysis in patients with stress-induced cardiomyopathy

Complexity-based analyses may quantify abnormalities in heart rate variability (HRV). The aim of this study was to investigate the clinical and prognostic significances of dynamic HRV changes in patients with stress-induced cardiomyopathy Takotsubo syndrome (TS) by means of linear and nonlinear analysis. Patients with TS were included in study after complete noninvasive and invasive cardiovascular diagnostic evaluation and compared to an age and gender matched control group of healthy subjects. Series of R–R interval and of ST–T interval values were obtained from 24-h ECG recordings after digital sampling. HRV analysis was performed by ‘range rescaled analysis’ to determine the Hurst exponent, by detrended fluctuation analysis to quantify fractal long-range correlation properties, and by approximate entropy to assess time-series predictability. Short- and long-term fractal-scaling exponents were significantly higher in patients with TS in acute phases, opposite to lower approximate entropy and Hurst exponent, but all variables normalized in a few weeks. Dynamic HRV analysis allows assessing changes in complexity features of HRV in TS patients during the acute stage, and to monitor recovery after treatment, thus complementing traditional ECG and clinically analysis.     

Assessment of Cytotoxic and Genotoxic Effect of Fissure Sealants in Buccal Epithelial Cells

AimThe main purpose of this study was to assess the genotoxic and cytotoxic effect of fissure sealants on buccal epithelial cells.Material and methodsThe study was conducted on 45 patients (27 girls and 18 boys), seven to 16 years of age (age mean 12.09 ± 2.20). Buccal swabs were collected before (T0), seven (T1), 30 (T2) and 90 days (T3) consequently after fissure sealant placement (Helioseal F®, Equia Fil®, Constic®). Patients or legal guardians filled in the questionnaire regarding the demographic data (age, gender), dietary habits, health status, medication usage, and recent X-ray exposure. DNA damage was analyzed using the micronucleus test.ResultsStatistically significant difference in the number of buccal cells with condensed chromatin was found between T0 (time before fissure sealant placement) and T3 (90 days after fissure sealant placement) period for Helioseal F® (P = 0.025). For the other two analyzed materials, no difference was observed during the tested period. There was no difference between materials in the same sampling time.ConclusionApart from an increase in cells with condensed chromatin 90 days after the placement of Helioseal F®, no other nuclear abnormalities were observed for tested fissure sealants. Although these sealants have now largely been used, it is of high importance that their biocompatibility is checked continuously, especially in in vivo clinical studies.