In Vivo Bio-Activation of JWH-175 to JWH-018: Pharmacodynamic and Pharmacokinetic Studies in Mice

3-(1-Naphthalenylmethyl)-1-pentyl-1H-indole (JWH-175) is a synthetic cannabinoid illegally marketed for its psychoactive cannabis-like effects. This study aimed to investigate and compare in vitro and in vivo pharmacodynamic activity of JWH-175 with that of 1-naphthalenyl (1-pentyl-1H-indol-3-yl)-methanone (JWH-018), as well as evaluate the in vitro (human liver microsomes) and in vivo (urine and plasma of CD-1 male mice) metabolic profile of JWH-175. In vitro binding studies showed that JWH-175 is a cannabinoid receptor agonist less potent than JWH-018 on mouse and human CB1 and CB2 receptors. In agreement with in vitro data, JWH-175 reduced the fESPS in brain hippocampal slices of mice less effectively than JWH-018. Similarly, in vivo behavioral studies showed that JWH-175 impaired sensorimotor responses, reduced breath rate and motor activity, and increased pain threshold to mechanical stimuli less potently than JWH-018. Metabolic studies demonstrated that JWH-175 is rapidly bioactivated to JWH-018 in mice blood, suggesting that in vivo effects of JWH-175 are also due to JWH-018 formation. The pharmaco-toxicological profile of JWH-175 was characterized for the first time, proving its in vivo bio-activation to the more potent agonist JWH-018. Thus, it highlighted the great importance of investigating the in vivo metabolism of synthetic cannabinoids for both clinical toxicology and forensic purposes.     

Evaluation of the antioxidant properties and oxidative stability of Pecorino cheese made from the raw milk of ewes fed Rosmarinus officinalis L. leaves

The aim of this work was to study the effect of rosemary leaf dietary supplementation on the antioxidant activities and total phenolic content of Pecorino cheese. Three hundred and twenty‐four sheep were randomly assigned to two dietary groups, which received a standard diet based on lucerne hay and concentrate (400 g per day). The concentrate of the rosemary supplemented group contained 2.50% dried rosemary leaves. The trial lasted 7 weeks. Cheesemaking was performed 3, 5 and 7 weeks from the start of the trial. The Pecorino cheese antioxidant activity was modified by the diet. Rosemary supplementation increased the total phenolic content, enhanced the antioxidant properties and decreased the lipid oxidation of cheese. A slight decrease in flavour was detected in cheeses after 7 weeks of rosemary administration.     

Different Apoptotic Pathways Activated by Oxaliplatin in Primary Astrocytes vs. Colo-Rectal Cancer Cells

Oxaliplatin-based chemotherapy improves the outcomes of metastatic colorectal cancer patients. Its most significant and dose-limiting side effect is the development of a neuropathic syndrome. The mechanism of the neurotoxicity is unclear. The limited knowledge about differences existing between neurotoxic and antitumor effects hinders the discovery of effective and safe adjuvant therapies. In vitro, we suggested cell-specific activation apoptotic pathways in normal nervous cells (astrocytes) vs. colon-cancer cells (HT-29). In the present research we compared the apoptotic signals evoked by oxaliplatin in astrocytes and HT-29 analyzing the intrinsic and extrinsic apoptotic pathways. In astrocytes, oxaliplatin induced a mitochondrial derangement measured as cytosolic release of cytochrome C, increase in superoxide anion levels and decreased expression of the antiapoptotic protein Bcl-2. Caspase-8, a main initiator of the extrinsic process remained unaltered. On the contrary, in HT-29 oxaliplatin increased caspase-8 activity and Bid expression, thus activating the extrinsic apoptosis, while the Bcl-2 increased expression blocked the mitochondrial damage. Data suggest the preferred activation of the intrinsic apoptosis as oxaliplatin damage signaling in normal nervous cells. The extrinsic pathway prevails in tumor cells indicating a possible strategy for planning new molecules to treat oxaliplatin-dependent neurotoxicity without negatively influence chemotherapy.     

Folic Acid Functionalized Surface Highlights 5‐Methylcytosine‐Genomic Content within Circulating Tumor Cells

Although the detection of methylated cell free DNA represents one of the most promising approaches for relapse risk assessment in cancer patients, the low concentration of cell‐free circulating DNA constitutes the biggest obstacle in the development of DNA methylation‐based biomarkers from blood. This paper describes a method for the measurement of genomic methylation content directly on circulating tumor cells (CTC), which could be used to deceive the aforementioned problem. Since CTC are disease related blood‐based biomarkers, they result essential to monitor tumor's stadiation, therapy, and early relapsing lesions. Within surface's bio‐functionalization and cell's isolation procedure standardization, the presented approach reveals a singular ability to detect high 5‐methylcytosine CTC‐subset content in the whole CTC compound, by choosing folic acid (FA) as transducer molecule. Sensitivity and specificity, calculated for FA functionalized surface (FA‐surface), result respectively on about 83% and 60%. FA‐surface, allowing the detection and characterization of early metastatic dissemination, provides a unique advance in the comprehension of tumors progression and dissemination confirming the presence of CTC and its association with high risk of relapse. This functionalized surface identifying and quantifying high 5‐methylcytosine CTC‐subset content into the patient's blood lead significant progress in cancer risk assessment, also providing a novel therapeutic strategy.     

Flux quantization effects in InN nanowires

InN nanowires, grown by plasma-enhanced molecular beam epitaxy, were investigated by means of magnetotransport. By performing temperature-dependent transport measurements and current measurements on a large number of nanowires of different dimensions, it is proven that the carrier transport mainly takes place in a tube-like surface electron gas. Measurements on three representative nanowires under an axially oriented magnetic field revealed pronounced magnetoconductance oscillations with a periodicity corresponding to a single magnetic flux quantum. The periodicity is explained by the effect of the magnetic flux penetrating the coherent circular quantum states in the InN nanowires, rather than by Aharonov-Bohm type interferences. The occurrence of the single magnetic flux quantum periodicity is attributed to the magnetic flux dependence of phase-coherent circular states with different angular momentum quantum numbers forming the one-dimensional transport channels. These phase coherent states can exist because of the almost ideal crystalline properties of the InN nanowires prepared by self-assembled growth.     

GABAB receptor intracellular trafficking after internalization in Paramecium

The number of neurotransmitter receptors on the plasma membrane is regulated by the traffic of intracellular vesicles. Golgi-derived vesicles provide newly synthesized receptors to the cell surface, whereas clathrin-coated vesicles are the initial vehicles for sequestration of surface receptors, which are ultimately degraded or recycled. We have previously shown that GABAB receptors display a punctuate vesicular pattern dispersed on the cell surface and throughout the cytoplasm and are internalized via clathrin-dependent and -independent endocytosis. Here we have studied constitutive GABAB receptor trafficking after internalization in Paramecium primaurelia by confocal laser scanning microscopy and multiple immunofluorescence analysis. After internalization, receptors are targeted to the early endosomes characterized by the molecular markers EEA1 and rab5. Some of these receptors, destined for recycling back to the plasma membrane, traffic from the early endosomes to the endosomal recycling compartment that is characterized by the presence of rab4-immunoreactivity (IR). Receptors that are destined for degradation exit the endosomal pathway at the early endosomes and traffic to the late endosome-lysosome pathway. In fact, some of the GABAB-positive compartments were identified as lysosomal structures by double staining with the lysosomal marker LAMP-1. GABAB vesicle structures also colocalize with TGN38-IR and rab11-IR. TGN38 and rab11 are proteins found in association with post-Golgi and recycling endosomes, respectively.     

Interplay between Structural and Thermoelectric Properties in Epitaxial Sb2+xTe3 Alloys

In recent years strain engineering is proposed in chalcogenide superlattices (SLs) to shape in particular the switching functionality for phase change memory applications. This is possible in Sb₂Te₃/GeTe heterostructures leveraging on the peculiar behavior of Sb₂Te₃, in between covalently bonded and weakly bonded materials. In the present study, the structural and thermoelectric (TE) properties of epitaxial Sb_2+xTe₃ films are shown, as they represent an intriguing option to expand the horizon of strain engineering in such SLs. Samples with composition between Sb₂Te₃ and Sb₄Te₃ are prepared by molecular beam epitaxy. A combination of X‐ray diffraction and Raman spectroscopy, together with dedicated simulations, allows unveiling the structural characteristics of the alloys. A consistent evaluation of the structural disorder characterizing the material is drawn as well as the presence of both Sb₂ and Sb₄ slabs is detected. A strong link exists among structural and TE properties, the latter having implications also in phase change SLs. A further improvement of the TE performances may be achieved by accurately engineering the intrinsic disorder. The possibility to tune the strain in designed Sb_2+xTe₃/GeTe SLs by controlling at the nanoscale the 2D character of the Sb_2+xTe₃ alloys is envisioned.     

Histatin-induced alterations in Candida albicans: a microscopic and submicroscopic comparison

Despite the numerous studies performed in an attempt to clarify the issue, the mechanism of action of salivary histatins remains unclear. The aim of the present study was to correlate histatin-induced morphological changes in Candida albicans by fluorescence microscopy (FM), transmission electron microscopy (TEM), and high resolution scanning electron microscopy (HRSEM). Each of the fluorescent dyes used by FM (i.e., tetramethylrhodamine methyl ester perchlorate for mitochondrial potential, Lysotracker for lysosome acidic compartment, and 4',6-diamino-2-phenylindole dihydrochloride for DNA) exhibited a specific staining in control cells. Following histatin treatment, we observed a recurring staining pattern, corresponding to fluorescence concentration along the cell periphery, suggesting a loss of dye specificity. To assess histatin-induced cytoplasmic modifications, ultrastructural analysis was then carried out. After treatments with histatins, TEM revealed characteristic intracellular modifications including: vacuole overgrowth, nuclear disappearance, loss of organelle identity, as well as the appearance of electron-dense membranes, likely of mitochondrial origin. Additionally, structures resembling autophagosomes were occasionally observed. By HRSEM, mitochondrial swelling was invariably the first sign of a histatin-induced effect. Other modifications included intracellular membrane disarrangement, organelles in disarray, and a large central cavity with deformed bodies displaced to the cell periphery, similar to what was detected by TEM. In summary, our study illustrates the occurrence of ultrastructural modifications following administration of histatins. Observations made with FM, TEM, and HRSEM provided different views of the same signs, demonstrating a definite action of histatins on C. albicans morphology. The possible functional meanings of these morphological results is discussed in light of the most recent biochemical data on histatin fungicidal activity.     

Long-term transport behavior of psychoactive compounds in sewage-affected groundwater

The present study provides a model-based characterization of the long-term transport behavior of five psychoactive compounds (meprobamate, pyrithyldione, primidone, phenobarbital and phenylethylmalonamide) introduced into groundwater via sewage irrigation in Berlin, Germany. Compounds are still present in the groundwater despite the sewage farm closure in the year 1980. Due to the limited information on (i) compound concentrations in the source water and (ii) substance properties, a total of 180 cross-sectional model realizations for each compound were carried out, covering a large range of possible parameter combinations. Results were compared with the present-day contamination patterns in the aquifer and the most likely scenarios were identified based on a number of model performance criteria. The simulation results show that (i) compounds are highly persistent under the present field conditions, and (ii) sorption is insignificant. Thus, back-diffusion from low permeability zones appears as the main reason for the compound retardation.