A Pharmacogenetics Study in Mozambican Patients Treated with Nevirapine: Full Resequencing of TRAF3IP2 Gene Shows a Novel Association with SJS/TEN Susceptibility

Steven–Johnson Syndrome (SJS) and Toxic Epidermal Necrolysis (TEN) are severe adverse drug reactions, characterized by extensive epidermal detachment and erosions of mucous membrane. SJS/TEN is one of the most serious adverse reactions to Nevirapine (NVP) treatment, commonly used in developing countries as first-line treatment of human immunodeficiency virus infection. In the last years TRAF3IP2 gene variants had been described as associated with susceptibility to several diseases such as psoriasis and psoriatic arthritis. We hypothesized that this gene, involved in immune response and in NF-κB activation, could also be implicated in the SJS/TEN susceptibility. We performed a full resequencing of TRAF3IP2 gene in a population of patients treated with NVP. Twenty-seven patients with NVP-induced SJS/TEN and 78 controls, all from Mozambique, were enrolled. We identified eight exonic and three intronic already described variants. The case/control association analysis highlighted an association between the rs76228616 SNP in exon 2 and the SJS/TEN susceptibility. In particular, the variant allele (C) resulted significantly associated with a higher risk to develop SJS/TEN (p = 0.012 and OR = 3.65 (95% CI 1.33–10.01)). A multivariate analysis by logistic regression confirmed its significant contribution (p = 0.027, OR = 4.39 (95% CI 1.19–16.23)). In conclusion, our study suggests that a variant in TRAF3IP2 gene could be involved in susceptibility to SJS/TEN.     

Effect of kaolin and copper based products and of starter cultures on green table olive fermentation

In the present study table olives treated in field with kaolin and copper based products against “olive-fruit fly” were fermented using two selected strains of lactic acid bacteria (LAB). The fermentation process was monitored up to 260 days from brining through physico-chemical, microbiological and sensorial analyses. Results showed a dominance of LAB and yeasts and low level of Enterobacteriaceae counts throughout the whole process both in un-treated and treated samples. When investigating the effect of the single treatments on microbial dynamics, ANOVA results highlighted that copper based products affected significantly the control sample, while the sample inoculated with LAB starters maintained high level throughout the process, guaranteeing the fermentation process. Different behavior was revealed by yeasts population, which was partially influenced by copper treatment at the beginning of the fermentation.The polyphasic approach used in the present study, which combined sensory evaluation to microbial counts and physico-chemical characteristics, let to the conclusion on the importance of starter cultures in fermentation of table olives especially those treated with “non-conventional” pesticide, which could be used to prevent olive fly damage.     

A one-pot ultrasound-assisted water extraction/cyclodextrin encapsulation of resveratrol from Polygonum cuspidatum

The ultrasound-assisted extraction (UAE) of resveratrol and other polyphenols from Polygonum cuspidatum has been carried out with the aim of developing an efficient and eco-friendly extraction process. The finely milled roots were sonicated (titanium horn, 19.5 kHz) in methanol, in different cyclodextrin water solutions [β-cyclodextrin (β-CD) or hydroxypropyl β-CD (HPβ-CD)] and also in pure water. UAE dramatically increased the yields and cut down extraction times compared to conventional extraction under stirring. Outstanding results have been achieved with the β-CD solution (1.5% w/w); in fact HPLC analysis showed that the selective inclusion properties of CDs toward phenolic stilbenes gave a much cleaner analytical extract profile. This green method gave 7.51 mg of total resveratrol (free + cleaved polydatin glucoside) per gram of dry plant. Thanks to polyphenol encapsulation within CDs, this extract showed excellent water dispersibility, higher stability and an antioxidant power which is comparable to that of the MeOH extract (DPPH, ORAC_FL). These important features should pave the road for its application in food supplements or phytochemical preparations.     

Low-enthalpy geothermal energy: An opportunity to meet increasing energy needs and reduce CO2 and atmospheric pollutant emissions in Piemonte,Italy

The scope for diffusion of very low-enthalpy geothermal plants in the Piemonte region of Italy, using groundwater heat pumps (GWHP), was analyzed to check environmental sustainability and the benefits in terms of reducing greenhouse gas emissions from fossil fuels. GWHP implementation seemed particularly suitable to the specific characteristics of the Piemonte plain. An important thick and productive shallow aquifer is present across the entire plain beneath the major energy users and is therefore appropriate for geothermal energy development purposes. The building stock could be adapted to heat pumps in different ways, but objective-oriented policies will be required to reach the best results in terms of environmental benefits.     

Plasma and recombinant thrombin-activable fibrinolysis inhibitor (TAFI) and activated TAFI compared with respect to glycosylation, thrombin/thrombomodulin-dependent activation, thermal stability, and enzymatic properties

Thrombin-activable fibrinolysis inhibitor (TAFI) is a human plasma zymogen similar to pancreatic pro-carboxypeptidase B. Cleavage of the zymogen by thrombin/thrombomodulin generates the enzyme, activated TAFI (TAFIa), which retards fibrin clot lysis in vitro and likely modulates fibrinolysis in vivo. In the present work we stably expressed recombinant TAFI in baby hamster kidney cells, purified it to homogeneity from conditioned serum-free medium, and compared it to plasma TAFI (pTAFI) with respect to glycosylation and kinetics of activation by thrombin/thrombomodulin. Although rTAFI is glycosylated somewhat differently than pTAFI, cleavage products with thrombin/thrombomodulin are indistinguishable, and parameters of activation kinetics are very similar with kcat = 0.55 s-1, K(m) = 0.54 microM, and Kd = 6.0 nM for rTAFI and kcat = 0.61 s-1, K(m) = 0.55 microM, and Kd = 6.6 nM for pTAFI. The respective TAFIa species also were prepared and compared with respect to thermal stability and enzymatic properties, including inhibition of fibrinolysis. The half-life of both enzymes at 37 degrees C is about 10 min, and the decay of enzymatic activity is associated with a quenching (to approximately 62% of the initial value at 60 min) of the intrinsic fluorescence of the enzyme. Stability was highly temperature-dependent, which, according to transition state theory, indicates both high enthalpy and entropy changes associated with inactivation (delta Ho++ approximately equal to 45 kcal/mol and delta So++ approximately equal to 80 cal/mol/K). Both species of TAFIa are stabilized by the competitive inhibitors 2-guanidinoethylmercaptosuccinic acid and epsilon-aminocaproic acid. rTAFIa and pTAFIa are very similar with respect to kinetics of cleavage of small substrates, susceptibility to inhibitors, and ability to retard both tPA-induced and plasmin-mediated fibrinolysis. These studies provide new insights into the thermal instability of TAFIa, a property which could be a significant regulator of its activity in vivo; in addition, they show that rTAFI and rTAFIa are excellent surrogates for the natural plasma-derived species, a necessary prerequisite for future studies of structure and function by site-specific mutagenesis.     

Radiolabeled Polymeric Nanoconstructs Loaded with Docetaxel and Curcumin for Cancer Combinatorial Therapy and Nuclear Imaging

Growing evidence suggests that multifaceted diseases as cancer can be effectively tackled by hitting simultaneously different biological targets and monitoring patient‐specific responses. Combinatorial therapies, relying on the administration of two or more molecules with different cytotoxic mechanisms, are rapidly progressing in the clinic. Here, 100 nm spherical polymeric nanoconstructs (SPNs) are proposed for the combinatorial treatment of tumors by codelivering a potent antimitotic drug—docetaxel (DTXL)—and a broad spectrum anti‐inflammatory molecule—curcumin (CURC). In vitro, SPNs loaded with DTXL and CURC induce a threefold decrease in IC₅₀ as compared to DTXL‐loaded SPNs. This synergic antitumor effect is also significant in mouse models of glioblastoma multiforme, where, after 22 d of treatment, the combinatorial approach leads to complete disease regression. At 90 d post‐treatment initiation, mice injected with DTXL + CURC SPNs have a 100% survival, whereas only 50% of the DTXL SPN treated mice survive. SPNs are also labeled with radioactive ⁶⁴Cu(DOTA) molecules to document, via PET imaging, the progressive tumor mass shrinkage. Sensitization of DTXL by CURC is associated with NF‐κB downregulation and increased apoptosis. These theranostic nanoconstructs could be used for combinatorial treatment and assessment of therapeutic efficacy in other malignancies.     

Tuning light emission of PbS nanocrystals from infrared to visible range by cation exchange

Colloidal semiconductor nanocrystals, with intense and sharp-line emission between red and near-infrared spectral regions, are of great interest for optoelectronic and bio-imaging applications. The growth of an inorganic passivation layer on nanocrystal surfaces is a common strategy to improve their chemical and optical stability and their photoluminescence quantum yield. In particular, cation exchange is a suitable approach for shell growth at the expense of the nanocrystal core size. Here, the cation exchange process is used to promote the formation of a CdS passivation layer on the surface of very small PbS nanocrystals (2.3 nm in diameter), blue shifting their optical spectra and yielding luminescent and stable nanostructures emitting in the range of 700–850 nm. Structural, morphological and compositional investigation confirms the nanocrystal size contraction after the cation-exchange process, while the PbS rock-salt crystalline phase is retained. Absorption and photoluminescence spectroscopy demonstrate the growth of a passivation layer with a decrease of the PbS core size, as inferred by the blue-shift of the excitonic peaks. The surface passivation strongly increases the photoluminescence intensity and the excited state lifetime. In addition, the nanocrystals reveal increased stability against oxidation over time. Thanks to their absorption and emission spectral range and the slow recombination dynamics, such highly luminescent nano-objects can find interesting applications in sensitized photovoltaic cells and light-emitting devices.     

Characterization of Pt/AlN/Pt-based structures for high temperature, microwave electroacoustic devices applications

Highly c-axis oriented AlN films, 3.15 μm thick, were grown by rf reactive sputtering technique at 200 °C on bare and Pt-covered Si(100) substrates previously oxidized to a thickness of about 2 μm in wet oxygen atmosphere. A Pt film, 2200 Å thick, was then sputtered on the free surface of the AlN/Pt/SiO₂/Si multilayer at 200 °C without breaking the vacuum in order to avoid any oxidation effects of the layers. The multilayers were then annealed in air at 900 °C for different time lengths up to 32 h in order to test the materials' resistivity to harsh environment. The influence of this high temperature annealing (HTA) on the thin films' crystallinity, as well as on the c-AlN piezoelectricity and Pt sheet resistivity was investigated at room temperature before and after each annealing. X ray diffraction investigations revealed that the films' crystallinity was improved by the HTA: the full width of half maximum of the AlN(002) and Pt(111) peaks decreases from 0.39° to 0.24°, and from 0.42° to 0.28° after 32-hours-HTA. Scanning electron microscopy, four points probe and piezoelectricity tests revealed that the morphology and the sheet resistivity (in the range from 0.6 to 0.5 Ω/sq) of the outer Pt film, as well as the AlN piezoelectric constants d₃₃ (in the range from 6.2 to 7.4⋅10⁻¹² C/N) was quite unaffected by the HTA even after 32 h of annealing.     

Hierarchically Structured Magnetic Nanoconstructs with Enhanced Relaxivity and Cooperative Tumor Accumulation

Iron oxide nanoparticles are formidable multifunctional systems capable of contrast enhancement in magnetic resonance imaging, guidance under remote fields, heat generation, and biodegradation. Yet, this potential is underutilized in that each function manifests at different nanoparticle sizes. Here, sub‐micrometer discoidal magnetic nanoconstructs are realized by confining 5 nm ultra‐small super‐paramagnetic iron oxide nanoparticles (USPIOs) within two different mesoporous structures, made out of silicon and polymers. These nanoconstructs exhibit transversal relaxivities up to ≈10 times (r ₂ ≈ 835 mm ^?1 s^?1) higher than conventional USPIOs and, under external magnetic fields, collectively cooperate to amplify tumor accumulation. The boost in r ₂ relaxivity arises from the formation of mesoscopic USPIO clusters within the porous matrix, inducing a local reduction in water molecule mobility as demonstrated via molecular dynamics simulations. The cooperative accumulation under static magnetic field derives from the large amount of iron that can be loaded per nanoconstuct (up to ≈65 fg) and the consequential generation of significant inter‐particle magnetic dipole interactions. In tumor bearing mice, the silicon‐based nanoconstructs provide MRI contrast enhancement at much smaller doses of iron (≈0.5 mg of Fe kg^?1 animal) as compared to current practice.     

Norepinephrine Protects against Methamphetamine Toxicity through β2-Adrenergic Receptors Promoting LC3 Compartmentalization

Norepinephrine (NE) neurons and extracellular NE exert some protective effects against a variety of insults, including methamphetamine (Meth)-induced cell damage. The intimate mechanism of protection remains difficult to be analyzed in vivo. In fact, this may occur directly on target neurons or as the indirect consequence of NE-induced alterations in the activity of trans-synaptic loops. Therefore, to elude neuronal networks, which may contribute to these effects in vivo, the present study investigates whether NE still protects when directly applied to Meth-treated PC12 cells. Meth was selected based on its detrimental effects along various specific brain areas. The study shows that NE directly protects in vitro against Meth-induced cell damage. The present study indicates that such an effect fully depends on the activation of plasma membrane β2-adrenergic receptors (ARs). Evidence indicates that β2-ARs activation restores autophagy, which is impaired by Meth administration. This occurs via restoration of the autophagy flux and, as assessed by ultrastructural morphometry, by preventing the dissipation of microtubule-associated protein 1 light chain 3 (LC3) from autophagy vacuoles to the cytosol, which is produced instead during Meth toxicity. These findings may have an impact in a variety of degenerative conditions characterized by NE deficiency along with autophagy impairment.