Role of the Endocannabinoid/Endovanilloid System in the Modulation of Osteoclast Activity in Paget’s Disease of Bone

The role of the endocannabinoid/endovanilloid (EC/EV) system in bone metabolism has recently received attention. Current literature evidences the modulation of osteoclasts and osteoblasts through the activation or inhibition of cannabinoid receptors in various pathological conditions with secondary involvement of bone tissue. However, this role is still unclear in primary bone diseases. Paget’s disease of the bone (PDB) could be considered a disease model for analyzing the role of the EC/EV system on osteoclasts (OCs), speculating the potential use of specific agents targeting this system for managing metabolic bone disorders. The aim of the study is to analyze OCs expression of EC/EV system in patients with PDB and to compare OCs activity between this population and healthy people. Finally, we investigate whether specific agents targeting EC/EV systems are able to modulate OCs activity in this metabolic bone disorder. We found a significant increase in cannabinoid receptor type 2 (CB2) protein expression in patients with PDB, compared to healthy controls. Moreover, we found a significant reduction in multi-nucleated tartrate-resistant acid phosphatase (TRAP)–positive OCs and resorption areas after treatment with JWH-133. CB2 could be a molecular target for reducing the activity of OCs in PDB, opening new therapeutic scenarios for the management of this condition.     

Identification and measurement of illicit drugs and their metabolites in urban wastewater by liquid chromatography-tandem mass spectrometry

Residues of illicit drugs and their metabolites that are excreted by humans may flow into and through wastewater treatment plants. The aim of this study was to develop a method for the determination of cocaine, amphetamines, morphine, cannabinoids, methadone, and some of their metabolites in wastewater. Composite 24-h samples from urban treatment plants were enriched with deuterated internal standards before solid-phase extraction. High-pressure liquid chromatography tandem mass spectrometry with multiple reaction monitoring was used for quantitation. Recoveries were generally higher than 80%, and limits of quantifications were in the low nanograms-per-liter range for untreated and treated wastewater. The overall variability of the method was lower than 10% for untreated and 5% for treated wastewater. The method was applied to wastewater samples coming from two treatment plants in Italy and Switzerland. Quantification ranges were found to be 0.2-1 microg/L for cocaine and its metabolite benzoylecgonine, 80-200 ng/L for morphine, 10 ng/L for 6-acetylmorphine, 60-90 ng/L for 11-nor-9-carboxy-Delta9-tetrahydrocannabinol, 10-90 ng/L for methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, and lower than 20 ng/L for amphetamines. As previously reported for cocaine, this method could be useful to estimate and monitor drug consumption in the population in real time, helping social scientists and authorities to combat drug abuse.     

The molecular mechanism of bee venom phospholipase A2 inactivation by bolinaquinone

The molecular basis of the inactivation of bee venom PLA2 by the marine natural product bolinaquinone (BLQ) was studied by several spectral techniques (CD, fluorescence, and NMR spectroscopy, mass spectrometry), biomimetic reactions, and molecular modeling. Our data suggest competitive inhibition based on a BLQ-PLA2 noncovalent molecular recognition. However, BLQ is also able to react selectively with Lys133 through conjugate addition followed by a beta elimination. The biological implications of both the covalent and noncovalent molecular events are discussed.     

The Antihypertensive Drug Telmisartan Protects Oligodendrocytes from Cholesterol Accumulation and Promotes Differentiation by a PPAR-γ-Mediated Mechanism

Our previous studies have demonstrated that specific peroxisome proliferator-activated receptor-γ (PPAR-γ) agonists play a fundamental role in oligodendrocyte progenitor (OP) differentiation, protecting them against oxidative and inflammatory damage. The antihypertensive drug Telmisartan (TLM) was shown to act as a PPAR-γ modulator. This study investigates the TLM effect on OP differentiation and validates its capability to restore damage in a pharmacological model of Niemann-Pick type C (NPC) disease through a PPAR-γ-mediated mechanism. For the first time in purified OPs, we demonstrate that TLM-induced PPAR-γ activation downregulates the type 1 angiotensin II receptor (AT1), the level of which naturally decreases during differentiation. Like other PPAR-γ agonists, we show that TLM promotes peroxisomal proliferation and promotes OP differentiation. Furthermore, TLM can offset the OP maturation arrest induced by a lysosomal cholesterol transport inhibitor (U18666A), which reproduces an NPC1-like phenotype. In the NPC1 model, TLM also reduces cholesterol accumulation within peroxisomal and lysosomal compartments and the contacts between lysosomes and peroxisomes, revealing that TLM can regulate intracellular cholesterol transport, crucial for myelin formation. Altogether, these data indicate a new potential use of TLM in hypomyelination pathologies such as NPC1, underlining the possible repositioning of the drug already used in other pathologies.     

Plasma Technology Increases the Efficacy of Prothioconazole against Fusarium graminearum and Fusarium proliferatum Contamination of Maize (Zea mays) Seedlings

The contamination of maize by Fusarium species able to produce mycotoxins raises great concern worldwide since they can accumulate these toxic metabolites in field crop products. Furthermore, little information exists today on the ability of Fusarium proliferatum and Fusarium graminearum, two well know mycotoxigenic species, to translocate from the seeds to the plants up to the kernels. Marketing seeds coated with fungicide molecules is a common practice; however, since there is a growing need for reducing chemicals in agriculture, new eco-friendly strategies are increasingly tested. Technologies based on ionized gases, known as plasmas, have been used for decades, with newer material surfaces, products, and approaches developed continuously. In this research, we tested a plasma-generated bilayer coating for encapsulating prothioconazole at the surface of maize seeds, to protect them from F. graminearum and F. proliferatum infection. A minimum amount of chemical was used, in direct contact with the seeds, with no dispersion in the soil. The ability of F. graminearum and F. proliferatum species to translocate from seeds to seedlings of maize has been clearly proven in our in vitro experiments. As for the use of plasma technology, the combined use of the plasma-generated coating with embedded prothioconazole was the most efficient approach, with a higher reduction of the infection of the maize seminal root system and stems. The debated capability of the two Fusarium species to translocate from seeds to seedlings has been demonstrated. The plasma-generated coating with embedded prothioconazole resulted in a promising sustainable approach for the protection of maize seedlings.     

Optical micromanipulations inside yeast cells

We present a combination of nonlinear microscopy and optical trapping applied to three-dimensional imaging and manipulation of intracellular structures in living cells. We use Titanium-sapphire laser pulses for nonlinear microscopy of the nuclear envelope and the microtubules marked with green fluorescent protein in fission yeast. The same laser source is also used to trap small lipid granules naturally present in the cell. The trapped granule is used as a handle to exert a pushing force on the cell nucleus. The granule is moved in a raster-scanning fashion to cover the area of the nucleus and hence displace the nucleus away from its normal position in the center of the cell. Such indirect manipulations of an organelle (e.g., nucleus) can be useful when direct trapping of the chosen organelle is disadvantageous or inefficient. We show that nonlinear microscopy and optical manipulation can be performed without substantial damage or heating of the cell. We present this method as an important tool in cell biology for manipulation of specific structures, as an alternative to genetic and biochemical methods. This technique can be applied to several fundamental problems in cell biology, including the mechanism of nuclear positioning and the spatial coordination of nuclear and cell division.     

Robotic techniques for upper limb evaluation and rehabilitation of stroke patients.

This paper presents two robot devices for use in the rehabilitation of upper limb movements and reports the quantitative parameters obtained to characterize the rate of improvement, thus allowing a precise monitoring of patient's recovery. A one degree of freedom (DoF) wrist manipulator and a two-DoF elbow-shoulder manipulator were designed using an admittance control strategy; if the patient could not move the handle, the devices completed the motor task. Two groups of chronic post-stroke patients (G1 n = 7, and G2 n = 9) were enrolled in a three week rehabilitation program including standard physical therapy (45 min daily) plus treatment by means of robot devices, respectively, for wrist and elbow-shoulder movements (40 min, twice daily). Both groups were evaluated by means of standard clinical assessment scales and a new robot measured evaluation metrics that included an active movement index quantifying the patient's ability to execute the assigned motor task without robot assistance, the mean velocity, and a movement accuracy index measuring the distance of the executed path from the theoretic one. After treatment, both groups improved their motor deficit and disability. In G1, there was a significant change in the clinical scale values (p < 0.05) and range of motion wrist extension (p < 0.02). G2 showed a significant change in clinical scales (p < 0.01), in strength (p < 0.05) and in the robot measured parameters (p < 0.01). The relationship between robot measured parameters and the clinical assessment scales showed a moderate and significant correlation (r > 0.53 p < 0.03). Our findings suggest that robot-aided neurorehabilitation may improve the motor outcome and disability of chronic post-stroke patients. The new robot measured parameters may provide useful information about the course of treatment and its effectiveness at discharge.     

Analysis and development of locomotion devices for the gastrointestinal tract

The authors are developing devices for semi-autonomous or autonomous locomotion in the gastrointestinal (GI) tract. In this paper, they illustrate the systematic approach to the problem of "effective" locomotion in the GI tract and the critical analysis of "inchworm" locomotion devices, based on extensor and clamper mechanisms. The fundamentals of locomotion and the practical problems encountered during the development and the testing (in vitro and in vivo) of these devices are discussed. A mini device capable of propelling itself in the colon and suitable to perform, at least, rectum-sigmoidoscopy (the tract where approximately 60% of all colon cancers are found) is presented. This paper introduces preliminary, but useful, concepts for understanding, modeling and improving the performance of virtually any existing and novel devices for endoscopy of the GI tract.     

Affinity capillary electrophoresis using a low-concentration additive with the consideration of relative mobilities

Most affinity studies in capillary electrophoresis assume that the analyte concentration is much smaller than the additive concentration so that the migration of the analyte has no effect on the concentration of the additive in the capillary. However, in most medium- to high-affinity interactions, the additive concentration has to be kept rather low to observe the changes in analyte mobility before saturation is reached. In this paper, a mathematical model is developed to describe the migration behavior of the analyte in a system where the complex formed becomes concentrated to levels much greater than the original concentration of the additive due to the differences in the mobilities of the analyte, additive, and complex. The analyte is flurbiprofen, the additive is transthyretin, and the stoichiometry of the reaction between the two is 1:2. This study also provides a new algorithm to determine medium- to high-affinity binding interaction constants by CE.     

Assessment of walking features from foot inertial sensing

An ambulatory monitoring system is developed for the estimation of spatio-temporal gait parameters. The inertial measurement unit embedded in the system is composed of one biaxial accelerometer and one rate gyroscope, and it reconstructs the sagittal trajectory of a sensed point on the instep of the foot. A gait phase segmentation procedure is devised to determine temporal gait parameters, including stride time and relative stance; the procedure allows to define the time intervals needed for carrying an efficient implementation of the strapdown integration, which allows to estimate stride length, walking speed, and incline. The measurement accuracy of walking speed and inclines assessments is evaluated by experiments carried on adult healthy subjects walking on a motorized treadmill. Root-mean-square errors less than 0.18 km/h (speed) and 1.52% (incline) are obtained for tested speeds and inclines varying in the intervals [3, 6] km/h and [-5, + 15]%, respectively. Based on the results of these experiments, it is concluded that foot inertial sensing is a promising tool for the reliable identification of subsequent gait cycles and the accurate assessment of walking speed and incline.