Betulinic Acid in Complex with a Gamma-Cyclodextrin Derivative Decreases Proliferation and in Vivo Tumor Development of Non-Metastatic and Metastatic B164A5 Cells

Betulinic acid, a very promising anti-melanoma agent, has very low water solubility that causes low bioavailability. To overcome this inconvenience, a highly water-soluble cyclodextrin was used (octakis-[6-deoxy-6-(2-sulfanyl ethanesulfonic acid)]-γ-cyclodextrin). The complex was physico-chemically analyzed using differential scanning calorimetry (DSC), X-ray and scanning electron microscopy (SEM) methods and then in vitro tested for its antiproliferative activity by the MTT assay and by cell cycle analysis. Finally, the complex was tested in vivo using an animal model of murine melanoma developed in C57BL/6J mice, where it caused a reduction in tumor volume and weight. The study revealed the beneficial influence of betulinic acid inclusion into the cyclodextrin in terms of antiproliferative activity and in vivo tumor development.     

pH and heat-induced structural changes of bovine apo-α-lactalbumin

The effects of pH and thermal treatments on conformation and association state of Ca²⁺-depleted bovine α-lactalbumin (apo-α-LA) have been studied by fluorescence spectroscopy, and molecular modelling. The experimental results demonstrate a third-state model for heat-induced unfolding of apo-α-LA, at pH 2.0, and an all-or-none transition of apo-α-LA, at pH 4.5 and 7.0, respectively. The heat-induced changes in the secondary and tertiary structure of α-LA were outlined after running molecular dynamics simulations at 25 °C and 80 °C, at neutral pH, therefore supporting the experimental observations. Our data provides insight into the mechanism of pH- and heat-dependent structural changes and oligomerization of α-LA, and will be helpful in understanding the ability of this protein to interact with certain compounds of biological interest.     

Unconventional method used in synthesis of polyphosphoesters

Polyphosphonates, polyphosphates, polyarylazophosphonates and polyarylazophosphates were synthesized by a new alternative to the classical phase transfer catalysis, respectively, the modified inverse phase transfer catalysis (IPTC) polycondensation of various phenylphosphonic (phosphoric) dichlorides (phenylphosphonic dichloride, phenylphosphoric dichloride, p-chlorophenylphosphoric dichloride) with bisphenols: bisphenol A, tetrabromobisphenol A, bis-(4-hydroxyphenyl)methane (bisphenol F), 4,4′-dihydroxyazobenzene. The polymers were characterized by infrared spectroscopy and magnetic resonance (¹H-NMR, ³¹P-NMR, ¹³C-NMR) spectroscopy. Yields in the range of 63.5–85% and molecular weights (M _w) of ~2,000–8,200 g mol^?1 were obtained. Polyphosphonates were stable up 210–270 °C and polyphosphates up 190–220 °C in air atmosphere. For a correct estimation of the thermal behavior of similar compounds, a kinetic analysis using a modified version of Non Parametric Kinetic method for representative polyphosphonate and polyphosphate was realized. The studies made on the hydrolytic degradation of the synthesized polyphosphates show that the most stable polymer under alkali-catalyzed degradation is the polyphosphate obtained by IPTC of phenylphosphoric dichloride and bisphenol A.     

Melanocytes Affect Nodal Expression and Signaling in Melanoma Cells: A Lesson from Pediatric Large Congenital Melanocytic Nevi

Expression of Nodal, a Transforming Growth Factor-beta (TGF-β) related growth factor, is associated with aggressive melanoma. Nodal expression in adult dysplastic nevi may predict the development of aggressive melanoma in some patients. A subset of pediatric patients diagnosed with giant or large congenital melanocytic nevi (LCMN) has shown increased risk for development of melanoma. Here, we investigate whether Nodal expression can help identify the rare cases of LCMN that develop melanoma and shed light on why the majority of these patients do not. Immunohistochemistry (IHC) staining results show varying degree of Nodal expression in pediatric dysplastic nevi and LCMN. Moreover, median scores from Nodal IHC expression analysis were not significantly different between these two groups. Additionally, none of the LCMN patients in this study developed melanoma, regardless of Nodal IHC levels. Co-culture experiments revealed reduced tumor growth and lower levels of Nodal and its signaling molecules P-SMAD2 and P-ERK1/2 when melanoma cells were grown in vivo or in vitro with normal melanocytes. The same was observed in melanoma cells cultured with melanocyte conditioned media containing pigmented melanocyte derived melanosomes (MDM). Since MDM contain molecules capable of inactivating radical oxygen species, to investigate potential anti-oxidant effect of MDM on Nodal expression and signaling in melanoma, melanoma cells were treated with either N-acetyl-l-cysteine (NAC), a component of the anti-oxidant glutathione or synthetic melanin, which in addition to providing pigmentation can also exert free radical scavenging activity. Melanoma cells treated with NAC or synthetic melanin showed reduced levels of Nodal, P-SMAD2 and P-ERK1/2 compared to untreated melanoma cells. Thus, the potential role for Nodal in melanoma development in LCMN is less evident than in adult dysplastic nevi possibly due to melanocyte cross-talk in LCMN capable of offsetting or delaying the pro-melanoma effects of Nodal via anti-oxidant effects of MDM.     

Comparative Study of 9×19 mm Ammunition Combustion Products and Residues

During the last decades, authorities’ awareness on environmental and human health impact at national and international level has increased in the field of small arms ammunition containing lead and antimony. Thus, the evolution of environmental policies regarding production and use of heavy metals and their compounds, especially in EU countries, implied extensive studies on the environmental implications of 9×19 mm caliber ammunition use, especially in indoor shooting ranges. In this context, the paper describes the experimental studies performed on Pb‐containing conventional FMJ (full metal jacketed) ammunition and comparative measurements on new ammunition designed for training. The combustion products and residues from conventional Pb‐containing ammunition and Cu‐based “greener” ammunition were compared in terms of gaseous products, metal concentration in aerosols and metal concentrations in solid residues. Gas emissions and solid residues were measured for four different types of ammunitions. The results showed that the copper content of the residue is significantly higher in case of Cu‐containing composite bullets. For the ammunition equipped with Pb bullet, an important percentage of the Pb residue is generated by erosion of the exposed part of the bullet core. On the other hand, the ammunition containing Cu‐composite bullet generates large quantities of Cu residues, which have similar environmental impact and less impact on human health. Gas analysis indicated the presence of CO and NO_x and, surprisingly, high concentrations of HCN, CH₄, and NH₃. Lower gas concentrations are obtained for the composite bullets due to an incomplete combustion of gun propellant.