Inhibition of melanin synthesis pathway by tricyclazole increases susceptibility of Fonsecaea pedrosoi against mouse macrophages

Fonsecaea pedrosoi produces melanin, a pigment related to virulence in pathogenic fungi. To understand the involvement of melanin in the protection of fungi, the authors used tricyclazole to inhibit the melanin pathway in F. pedrosoi. Experiments of pigmentation suggested that F. pedrosoi uniquely produces dihydroxynaphthalene-melanin. Pigments produced on cultures modified or not with tricyclazole were extracted by an alkali-acid method and submitted to infrared and ion exchange chromatography analysis; also cytochemistry analysis for cationized ferritin of whole cells was carried out. This group of experiments showed that the tricyclazole treatment on F. pedrosoi produced a melanin-like pigment, but less negatively charged and with less affinity for iron ions than that without the tricyclazole treatment, and this in turn lead to a less negatively charge cell wall surface. Scanning electron microscopy of such pigments showed that the melanin from control cultures maintained their hyphae-like structures, which have been described as "melanin-ghosts," whereas the tricyclazole pigment showed an amorphous surface. Interaction of conidia from cultures of F. pedrosoi, modified by tricyclazole or not, with peritoneal activated macrophages suggested that tricyclazole causes higher association of fungus with macrophages, weakens the fungus capacity to destroy the macrophages, and diminishes the resistance to dry fracture procedures on samples prepared for high resolution scanning electron microscopy.     

alpha,alpha-Cyclopentaneglycine dipeptides capped with biaryls as tachykinin NK(2) receptor antagonists

The NK(2) receptor belongs to the family of tachykinin neurotransmitters. It has been reported to be involved in several pathological conditions, and selective antagonists are potentially useful drugs for the treatment of asthma, irritable bowel syndrome, cystitis, and depression. Starting from in-house capped dipeptide libraries, we were able to identify a number of molecules with sub-nanomolar binding affinity for the hNK(2) receptor. All were characterized by a rigid core structure with a strong constraint induced by an alpha,alpha-cyclopentaneglycine fragment. Herein we report the further elaboration of three initial basic structures. The planar benzothiophene group was substituted with a series of biphenyl and heterobiphenyl moieties that are well tolerated in terms of receptor affinity. The new compounds also maintained good antagonist potency in an in vitro functional assay, and a number of them showed significant in vivo activity after intravenous administration in our guinea pig model.     

Effect of cooling rate on melt-crystallization of random propylene-ethylene and propylene-1-butene copolymers

Summary  The effect of variation of the rate of cooling on melt-crystallization of random copolymers of isotactic polypropylene with low amount of either ethylene or 1-butene is evaluated using X-ray techniques, atomic force microscopy, and calorimetry. Slow non-isothermal melt-crystallization mainly results in formation of monoclinic α-crystals of lamellar geometry. The presence of comonomers leads to a decrease of the degree of crystallinity, and of the thickness of lamellae. These changes are proportional to the concentration of co-units, and are more distinct in propylene-ethylene copolymers. Rapid cooling of propylene-1-butene copolymers leads to formation of non-lamellar mesomorphic domains, independent of the concentration of co-units within the investigated range of concentrations. In quenched propylene-ethylene copolymers, in contrast, the formation of mesomorphic structure partially is replaced by formation of monoclinic α-crystals of still non-lamellar geometry.     

Hyaluronidase Alters the Lipid Profile of Cumulus Cells as Detected by MALDI-TOF MS and Multivariate Analysis

This research aimed to study the changes in lipid composition in cumulus cells using hyaluronidase according to the intracytoplasmic sperm injection protocol commonly used in human reproduction clinics. The lipid extraction was performed by the Blight‐Dyer protocol and the lipid profiles were obtained by MALDI‐TOF MS in positive and negative modes. The mass spectra data were processed with MassLynx and the statistical analysis was performed using MetaboAnalyst 2.0. Fifteen ions were selected for each mode as potential markers for differences between the groups. These ions were identified in the human metabolome database as phosphatidylserine with and without treatment, phosphatidylethanolamine in the after treatment group and phosphatidylinositol in the before treatment group, which are lipids that may be involved in cell apoptosis and signaling. We concluded that MALDI‐TOF MS coupled with multivariate analysis can be utilized as a strategy to obtain and study the lipid profiles of cumulus cells and as a tool to study the metabolic state of cumulus cells.     

A Bisbenzamidine Phosphonate as a Janus‐faced Inhibitor for Trypsin‐like Serine Proteases

A hybrid approach was applied for the design of an inhibitor of trypsin‐like serine proteases. Compound 16 [(R,R)‐ and (R,S)‐diphenyl (4‐(1‐(4‐amidinobenzylamino)‐1‐oxo‐3‐phenylpropan‐2‐ylcarbamoyl)phenylamino)(4‐amidinophenyl)methylphosphonate hydrochloride], prepared in a convergent synthetic procedure, possesses a phosphonate warhead prone to react with the active site serine residue in a covalent, irreversible manner. Each of the two benzamidine moieties of 16 can potentially be accommodated in the S1 pocket of the target enzyme, but only the benzamidine close to the phosphonate group would then promote an irreversible interaction. The Janus‐faced inhibitor 16 was evaluated against several serine proteases and caused a pronounced inactivation of human thrombin with a second‐order rate constant (k_inac/K_i) of 59 500 M ^?1 s^?1. With human matriptase, 16 showed preference for a reversible mode of inhibition (IC₅₀=2.6 μM ) as indicated by linear progress curves and enzyme reactivation.     

Synthesis of Gd and (68)Ga complexes in conjugation with a conformationally optimized RGD sequence as potential MRI and PET tumor-imaging probes

We report the synthesis of novel chelates of Gd and (68)Ga with DPTA, DOTA, HP-DOA3, as well as with AAZTA, a novel chelating agent developed by our research group. These chelating agents were appropriately conjugated, prior to metal complexation, with DB58, an RGD peptidomimetic, conformationally constrained on an azabicycloalkane scaffold and endowed with high affinity for integrin α(ν)β(3) . Because α(ν)β(3) is involved in neo-angiogenesis in solid tumors and is also directly expressed in cancer cells (e.g. glioblastomas, melanomas) and ovarian, breast, and prostate cancers, these constructs could prove useful as molecular imaging probes in cancer diagnosis by MRI or PET techniques. Molecular modeling, integrin binding assays, and relaxivity assessments allowed the selection of compounds suitable for multiple expression on dendrimeric or nanoparticulate structures. These results also led us to an exploratory investigation of (68)Ga complexation for the promising (68)Ga-PET technique; the AAZTA complex 15((68)Ga) exhibited uptake in a xenograft model of glioblastoma, suggesting potentially useful developments with new probes with improved affinity.     

Plant Volatiles Enhance Behavioral Responses of Grapevine Moth Males, Lobesia botrana to Sex Pheromone

Plant volatiles play an important role in the lives of phytophagous insects, by guiding them to oviposition, feeding and mating sites. We tested the effects of different host-plant volatiles on attraction of Lobesia botrana males to the female-produced sex pheromone, in a wind tunnel. Addition of volatile emissions from grapevines or individual plant volatiles to pheromone increased the behavioral responses of L. botrana males over those to pheromone alone. At a low release rate (under-dosed) of pheromone, addition of (E)-β-caryophyllene, (Z)-3-hexenyl acetate, 1-hexanol, or 1-octen-3-ol increased all behavioral responses, from activation to pheromone source contact, while addition of (E)-4,8-dimethyl-1,3,7-nonatriene, (E)-β-farnesene, (Z)-3-hexenol, or methyl salicylate affected only the initial behavioral responses. Dose–response experiments suggested an optimal release ratio of 1:1000 (sex pheromone: host plant volatile). Our results highlight the role of plant volatiles in the sensory ecology of L. botrana.     

Mixing and Reactions in Microchannels – an Educational Approach Using the Internet

In many fields, microprocess technology is gaining significance and is increasingly finding its way into use in industrial production. However, specialist knowledge regarding reactions of compounds, mixing and downstream processes in microchannels is required, as experience from the ”?macro world”? can only be applied in a limited way. The appropriate level of education required is still lacking. In this study, a number of educational experiments were set up focusing on mixing processes and emphasizing the significance of viscosity in mixing. Additionally liquid‐liquid extraction as an important downstream process was looked into as a student experiment. In order to make the training widely accessible, a web‐based education platform was established.     

Evaluation of nonconventional additives as fire retardants on polyamide 6,6: Phosphorous‐based master batch, α‐zirconium dihydrogen phosphate,and β‐cyclodextrin based nanosponges

The influence of new types of additives, such as halogen‐ and antimony‐free flame‐retardant master batches based on phosphorus, α‐zirconium dihydrogen phosphate, and β‐cyclodextrin nanosponges, on the flame retardancy of polyamide 6,6 (PA6,6) by means of cone calorimetry and limiting oxygen index (LOI) tests was investigated. A significant decrease of the heat release rate, depending by the type of additive used, was observed. Furthermore, with the consideration that the life safety during the fire could be improved by a decrease in the fire hazard, a decrease in the quantity of the smoke and its toxicity, depending also on the type of additive, was revealed. With regard to the LOI test, neat PA6,6 showed a slight increase in the LOI value in comparison with the PA6,6 composites. However, all of the PA6,6/composites showed a slower burning velocity and antidripping effects at oxygen concentrations corresponding to the LOI value. To understand the flame‐retardancy mechanism of these novel PA6,6 composites, we thoroughly investigated their thermal decomposition behavior and microstructure/elemental analysis by scanning electron microscopy/energy‐dispersive X‐ray spectroscopy. Furthermore, the combustion behavior of these novel PA6,6 composites was compared with that of conventional nanofillers (e.g., modified montmorillonite clay and carbon nanotubes). © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012