In vitro photodynamic activity of chloro(5,10,15,20-tetraphenylporphyrinato)indium(III) loaded-poly(lactide-co-glycolide) nanoparticles in LNCaP prostate tumour cells

In(III)-meso-tetraphenylporphyrin (InTPP) was encapsulated into nanoparticles (smaller than 200 nm) of poly(d,l-lactide-co-glycolide) (PLGA) using the emulsification–evaporation technique. The photodynamic efficacy of InTPP-loaded nanoparticles and its cellular uptake was investigated with LNCaP prostate tumour cells, in comparison with the free InTPP. The effects of incubation time (1–3 h), drug concentration (1.8–7.7 μmol/L) and incident light dose (15–45 J/cm²) with both encapsulated and free InTPP were studied. The type of cell death induced by the photochemical process using both encapsulated and free InTPP was also investigated. Cell viability was reduced more significantly with increasing values of these effects for InTPP-loaded nanoparticles than with the free drug. The cellular death induced by both encapsulated and free InTPP was preponderantly apoptotic. Confocal laser scanning microscopy data showed that the InTPP-loaded nanoparticles, as well free InTPP, were localized in the cells, and always in the perinuclear region. Encapsulated InTPP was measured by the intensity of fluorescence intensity of cell extracts and was three times more internalized into the cells than was the free InTPP. Electron paramagnetic resonance experiments corroborated the participation of singlet oxygen in the photocytotoxic effect of nanoparticles loaded with InTPP.     

Modified and Optimized Glass Electrode for pH Measurements in Hydrated Ethanol Fuel

One of the quality control parameters of ethanol fuel is pH, established by the Brazilian standard ABNT NBR 10891, whose scope is specific for hydrated ethanol fuel, and by the American standard ASTM D 6423, which focuses on anhydrous ethanol fuel. This study presented a modified and optimized structure using a single solvent, both for the glass electrode and the external reference electrode, to minimize the presence of the liquid junction potential for measuring the pH of hydrated ethanol fuel. The Box–Behnken design enabled us to determine the optimal condition expected for the new measurement system, which was compared with the systems proposed by the standard references and the turning range of acid–base indicators using parametric and nonparametric tests. The results revealed that the pH values obtained by the different systems are statistically different, and that only the values obtained by this proposal are suitable for the pH range found by the indicators. The optimized electrode presented an adequate response sensitivity to the Nernst equation, having an operational behavior adequate for the modified and optimized glass electrode for pH measurements in hydrated ethanol fuel.     

Targeting retroviral Zn finger-DNA interactions: a small-molecule approach using the electrophilic nature of trans-platinum-nucleobase compounds

Noncovalent interactions are ubiquitous in ternary systems involving metal ions, DNA/RNA, and proteins and represent a structural motif for design of selective inhibitors of biological function. This contribution shows that small molecules containing platinated purine nucleobases mimic the natural DNA(RNA)-tryptophan recognition interaction of zinc finger peptides, specifically the C-terminal finger of HIV NCp7 protein. Interaction with platinum results in Zn ejection from the peptide accompanied by loss of tertiary structure. Targeting the NCp7-DNA interaction for drug design represents a conceptual advance over electrophiles designed for chemical attack on the zinc finger alone. These results demonstrate examples of a new platinum structural class targeting specific biological processes, distinct from the bifunctional DNA-DNA binding of cytotoxic agents like cisplatin. The results confirm the validity of a chemical biological approach for metallodrug design for selective ternary DNA(RNA)-protein interactions.     

Stability of controlled inverted pendulum under permanent horizontal perturbations of the supporting point

We consider the problem of choosing a test perturbation of a movable foundation of a single-link inverted pendulum so as to test a vestibular prosthesis prototype located at the top of this pendulum in an extreme situation. The obtained results permit concluding that the information transmitted from otolithic organs of the human vestibular system to muscles of the locomotor apparatus is very important and improves the quality of stabilization of the human vertical posture preventing the possible fall.     

A Novel β-Glucosidase from Humicola insolens with High Potential for Untreated Waste Paper Conversion to Sugars

Humicola insolens produced a new β-glucosidase (BglHi2) under solid-state fermentation. The purified enzyme showed apparent molecular masses of 116 kDa (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) and 404 kDa (gel-filtration), suggesting that it is a homotetramer. Mass spectrometry analysis showed amino acid sequence similarity with a β-glucosidase from Chaetomium thermophilum. Optima of pH and temperature were 5.0 and 65 °C, respectively, and the enzyme was stable for 60 min at 50 °C, maintaining 71 % residual activity after 60 min at 55 °C. BglHi2 hydrolyzed p-nitrophenyl-β-d-glucopyranoside and cellobiose. Cellobiose hydrolysis occurred with high apparent affinity (K _M?=?0.24?±?0.01 mmol L^?1) and catalytic efficiency (k _cat/K _M?=?1,304.92?±?53.32 L mmol^?1 s^?1). The activity was insensitive to Fe⁺³, Cr⁺², Mn⁺², Co⁺², and Ni²⁺, and 50–60 % residual activities were retained in the presence of Pb²⁺, Hg²⁺, and Cu²⁺. Mixtures of pure BglHi2 or H. insolens crude extract (CE) with crude extracts from Trichoderma reesei fully hydrolyzed Whatman no. 1 paper. Mixtures of H. insolens CE with T. reesei CE or Celluclast 1.5 L fully hydrolyzed untreated printed office paper, napkin, and magazine papers after 24–48 h, and untreated cardboard was hydrolyzed by a H. insolens CE/T. reesei CE mixture with 100 % glucose yield. Data revealed the good potential of BglHi2 for the hydrolysis of waste papers, promising feedstocks for cellulosic ethanol production.     

Multi-Dimensional Structure and Dynamics Landscape of Proteins in Mammalian Cells Revealed by In-Cell NMR

Governing function, half-life and subcellular localization, the 3D structure and dynamics of proteins are in nature constantly changing in a tightly regulated manner to fulfill the physiological and adaptive requirements of the cells. To find evidence for this hypothesis, we applied in-cell NMR to three folded model proteins and propose that the splitting of cross peaks constitutes an atomic fingerprint of distinct structural states that arise from multiple target binding co-existing inside mammalian cells. These structural states change upon protein loss of function or subcellular localisation into distinct cell compartments. In addition to peak splitting, we observed NMR signal intensity attenuations indicative of transient interactions with other molecules and dynamics on the microsecond to millisecond time scale.     

New Cholesteric and Nematic Lyotropic Mesophases from Di-Sodium N-Lauroyl-Aspartate

Lyotropic liquid crystals are prepared from racemic and optically active di-sodium N-lauroyl aspartate (SNLA). The optically active levo enantiomer is a suitable amphiphile for obtaining cholesteric lyotropic mesophases. In the present work, different types of mesophases, including cholesteric and nematic, were prepared and their type I or II magnetic behavior characterized by deuterium and ²³Na NMR. The polarizing microscope textures were investigated, particularly with regards to magnetic field effects upon the samples. The usual chevron pattern was obtained for particular sample orientations and several characteristic effects were observed for the type I cholesteric lyomesophase in the first few minutes following removal of the sample from the magnetic field. Although the head group of di-sodium N-lauroyl aspartate possesses two negative charges, compositions are not too different from those reported for other amphiphile lyotropic systems.     

Screening of pesticides via solid-phase extraction and gas chromatography

This work describes an analytical method for the screening of pesticide residues in water. The developed method identifies and quantitates a variety of pesticides in water samples such as organochlorine, organophosphate, chloronitrile, phthalimide, dicarboximide, and triazine. The analytical technique employed for the extraction and cleanup step involves solid-phase extraction with C-18 cartridges. The identification and determination of concentration is carried out with gas chromatography. Two columns of different polarity and two specific detectors are used. An electron capture detector is used for organochrolorine, and a thermoinic detector is used for organonitrogen and organophosphate. The good resolution achieved with the temperature program and the combination of columns is shown in the chromatograms obtained. The limits of detection, percent recovery, and their respective standard deviations are determined for each product. According to the results obtained, this method permits monitoring of the pesticides mentioned, many of which are moderately to highly toxic to aquatic organisms.     

Covalent and noncovalent interactions for [metal(dien)nucleobase](2+) complexes with L-tryptophan derivatives: formation of palladium-tryptophan species by nucleobase substitution under biologically relevant conditions

The interaction of the complexes [Pd(dien)(1-MeCyt)]2+ (2) and [Pd(dien)(9-EtGH)]2+ (3) with the amino acids L-tryptophan (Trp) and N-acetyltryptophan (N-AcTrp) was studied and compared with the previously studied platinum analogues [Pt(dien)(1-MeCyt)]2+ (4) and [Pt(dien)(9-EtGH)]2+ (5). Solid-state structures for 2 and 4 are reported. For the palladium complexes, the interaction is pH sensitive. Below pH 5, the noncovalent interaction with stacking between the aromatic amino acid residue and the metalated nucleobase was observed. Fluorescence quenching experiments indicated similar association constants for platinum and palladium derivatives 2-5. Unusual substitution of the model nucleobases 1-methylcytosine (1-MeCyt) and 9-ethylguanine (9-EtGH) by tryptophan was observed in the range of pH 5-11. The resulting species [Pd(dien)(Trp)]+ (6) and [Pd(dien)(N-AcTrp)]+ (7) were characterized using 1H NMR, 13C NMR, and ESI-MS spectroscopy with coordination indicated through the amino and deprotonated amido nitrogens, respectively. Complexes 6 and 7 were also obtained from a solution of [Pd(dien)Cl]+ (1) incubated with either Trp or N-AcTrp, respectively.