Methodology for characterization of platinum-based drug's targeted delivery nanosystems.

Conventional anticancer therapies exploiting platinum-based drugs rely principally on the intravascular injection of the therapeutic agent. The anticancer drug is distributed throughout the body by the systemic blood circulation undergoing cellular uptake, rapid clearance and excretion. Consequently, only a small portion of the platinum-based drug reaches the tumor site, which is associated with severe side effects. For this reason, targeted delivery systems are of great need since they offer enhanced and selective delivery of a drug to cancerous cells making the therapy safe and more effective. Up to date, a variety of the Pt-based drug targeted delivery systems (Pt-based DTDSs) utilizing nanomaterials have been developed and tested using a range of analytical techniques that provided essential information on their synthesis, stability, biodistribution and cytotoxicity. Here we summarize those experimental techniques indicating their applicability at different stages of the research, as well as pointing out their strengths, advantages, drawbacks and limitations. Also, the existing strategies and approaches are critically reviewed with the objective to reveal and give rise to the development of the analytical methodology suitable for reliable Pt-based DTDSs characterization which would eventually result in novel therapies and better patients' outcomes.

Isotonic Saline Nebulization and Lung Function in Children With Mild Respiratory Ailments.

Nebulization with saline solution, although commonly used to alleviate respiratory symptoms, particularly in children, is often questioned concerning its effectiveness. In this study, we investigated the effects of isotonic saline nebulization on lung function in 40 children (mean age of 14±1 years) suffering from different types of airway disorders. Measurements were carried out directly before and up to 15 min after nebulization, for six days in a row, always on the same day time in the morning. The children were divided into two study groups according to the baseline ratio of forced expired volume in one second/forced vital capacity (FEV1/FVC), below and above 80 %. We found significant improvements after saline nebulization in FEV1, mid-expiratory flow at 50 % and 75 % of FVC (MEF50 and MEF75), and peak expiratory flow (PEF) in the group with the baseline FEV1/FVC less than 80 %. In contradistinction, children with an index greater than 80 % displayed no appreciable changes in the lung function variables when compared with the baseline level before saline nebulization. We conclude that isotonic saline nebulization might mitigate the functional signs of threatening pulmonary obstruction and as such may be clinically useful in pediatric patients with mild respiratory problems.

Study of the absorption coefficient of graphene-polymer composites.

In this work, we have prepared a series of polydimethylsiloxane (PDMS) composites containing various graphene flakes loadings (0.02-2 wt%), and their broadband optical properties are being investigated. We demonstrate the tunability and evolution of transmittance and reflection spectra of the composites in a wide spectral range (0.4-200 µm) as a function of graphene content. Using these data we derive the broadband wavelength-dependent absorption coefficient (α) values. Our results show that α is roughly constant in the visible and IR ranges, and, surprisingly, is approximately one order of magnitude lower in the terahertz regime, suggesting different terahertz radiation scattering mechanism in our composite. Our material could be useful for applications in optical communication, sensing or ultrafast photonics.

Statistical analysis of the reduction process of graphene oxide probed by Raman spectroscopy mapping.

We propose a method for monitoring the large-scale homogeneity of the reduction process of graphene oxide. For this purpose, a Raman mapping technique is employed to probe the evolution of the phonon properties of two different graphene oxide (GO) thin films upon controllable thermal reduction. The reduction of GO is reflected by the upshift of the statistical distribution of the relative intensity ratio of the G and D peaks (I D/I G) of the Raman spectra and is consistent with the ratio obtained for chemically reduced GO. In addition, the shifts of the position distributions of the main Raman modes ([Formula: see text], [Formula: see text]) and their cross-correlation with the I D/I G ratio provides evidence of a change of the doping level, demonstrating the influence of reduction processes on GO films.

Enhanced uptake of blood coagulation factor VIII containing immune complexes by antigen presenting cells.

Essentials Anti-factor (F) VIII antibody formation is a major complication in the treatment of hemophilia A. We investigated uptake of FVIII and FVIII immune complex by bone marrow derived dendritic cells. Immune complex formation increased uptake of FVIII 3-4 fold in a Fcγ receptor dependent manner. FVIII immune complex binding to Fcγ receptors may modulate immune tolerance induction. SUMMARY: Background A major complication in the treatment of hemophilia A is the development of inhibitory antibodies targeting coagulation factor VIII (FVIII). Eradication of these inhibitors can be established by immune tolerance induction (ITI), which consists of daily administration of high dosages of FVIII. FVIII immune complexes (FVIII-IC) could be formed following FVIII infusion in patients with pre-existing anti-FVIII antibodies. Objectives Here we studied endocytosis of FVIII-IC by bone marrow-derived dendritic cells (BMDCs). Methods BMDCs were pulsed with FVIII/FVIII-IC and uptake was assessed by flow cytometry and confocal imaging. Results BMDCs were able to efficiently internalize FVIII-IC in a dose-dependent manner, 3-4-fold more efficiently when compared with equimolar concentrations of non-complexed FVIII. Uptake of FVIII-IC, but not FVIII alone, could be inhibited with anti-Fcγ receptor (FcγR) antibody 2.4G2, indicating functional involvement of FcγR. No internalization of FVIII-IC was observed in BMDCs lacking FcγRI, FcγRIIb, FcγRIII and FcγRIV. Genetic ablation of FcγRIIb, FcγRIII or FcγRIV individually did not affect the ability of anti-FVIII IgG to promote the uptake of FVIII. BMDCs lacking FcγRI showed lower FVIII-IC uptake levels when compared with other single FcγR null BMDCs. Expression of the inhibitory FcγRIIb alone was sufficient to internalize FVIII-IC more efficiently than FVIII. Conclusions FcγR are critical in the internalization of FVIII-IC by BMDCs and multiple FcγR can contribute independently to this process. Our findings provide a basis for future studies to address whether the outcome of ITI is dependent on the interplay between FVIII-IC and inhibitory and activating FcγR.

Dangerous liaisons: how the immune system deals with factor VIII.

Only a fraction of patients with hemophilia A develop a neutralizing antibody (inhibitor) response to therapeutic infusions of factor VIII. Our present understanding of the underlying causes of the immunogenicity of this protein is limited. In the past few years, insights into the uptake and processing of FVIII by antigen-presenting cells (APCs) have expanded significantly. Although the mechanism of endocytosis remains unclear, current data indicate that FVIII enters APCs via its C1 domain. Its subsequent processing within endolysosomes allows for presentation of a heterogeneous collection of FVIII-derived peptides on major histocompatibility complex (MHC) class II, and this peptide-MHC class II complex may then be recognized by cognate effector CD4(+) T cells, leading to anti-FVIII antibody production. Here we aim to summarize recent knowledge gained about FVIII processing and presentation by APCs, as well as the diversity of the FVIII-specific T-cell repertoire in mice and humans. Moreover, we discuss possible factors that can drive FVIII immunogenicity. We believe that increasing understanding of the immune recognition of FVIII and the cellular mechanisms of anti-FVIII antibody production will lead to novel therapeutic approaches to prevent inhibitor formation in patients with hemophilia A.

1H and 13C NMR spectra, structure and physicochemical features of phenyl acridine-9-carboxylates and 10-methyl-9-(phenoxycarbonyl)acridinium trifluoromethanesulphonates--alkyl substituted in the phenyl fragment.

The 1H and 13C NMR spectra of twelve phenyl acridine-9-carboxylates--alkyl-substituted in the phenyl fragment--and their 10-methyl-9-(phenoxycarbonyl)acridinium salts dissolved in CD3CN, CD3OD, CDCl3 and DMSO-d6 were recorded in order to examine the influence of the structure of these compounds and the properties of the solvents on chemical shifts and 1H-(1)H coupling constants. Experimental data were compared with 1H and 13C chemical shifts predicted at the GIAO/DFT level of theory for DFT(B3LYP)/6-31G** optimised geometries of molecules, as well as with values of 1H chemical shifts and 1H-(1)H coupling constants, estimated using ACD/HNMR database software to ensure that the assignment was correct. To investigate the relations between chemical shifts and selected structural or physicochemical characteristics of the target compounds, the values of several of these parameters were determined at the DFT or HF levels of theory. The HOMO and LUMO energies obtained at the HF level yielded the ionisation potentials and electron affinities of molecules. The DFT method provided atomic partial charges, dipole moments, LCAO coefficients of pz LUMO of selected C atoms, and angles reflecting characteristic structural features of the compounds. It was found that the experimentally determined 1H and 13C chemical shifts of certain atoms relate to the predicted dipole moments, the angles between the acridine and phenyl moieties, and the LCAO coefficients of the pz LUMO of the C atoms believed to participate in the initial step of the oxidation of the target compounds. The spectral and physicochemical characteristics of the target compounds were investigated in the context of their chemiluminogenic ability.

Phenotypic and evolutionary plasticity of body composition in rats selectively bred for high endurance capacity.

We investigated the effects of genetic selection and prolonged wheel access (8 wk) on food consumption and body composition in lines of rats selected for high and low intrinsic (untrained) endurance running capacity (HCR and LCR, respectively) to test the generality of phenotypic correlations between physical activity levels, aerobic capacity, and body composition. HCR rats ran more minutes per day on activity wheels than LCR rats, supporting the hypothesis that voluntary activity and physiological capacity are genetically correlated (self-induced adaptive plasticity). Both treatments (selection and wheel access) significantly affected food consumption. HCR rats consumed and digested more food than LCR rats. Access to running wheels did not result in changes in overall body mass, but lean body mass increased and percent body fat decreased in both lines. Selection for high endurance capacity resulted in hypertrophy of the heart and kidneys and decreased long intestine length. We found significant phenotypic flexibility in a number of organ masses after wheel running. Specifically, access to running wheels resulted in hypertrophy of the heart, liver, kidney, stomach, and small and large intestines in LCR and HCR rats. The selected line×wheel access interaction was significantly greater in HCR rats in relative mass for the heart and lung. Compared with LCR rats, HCR rats fortify wheel running with increased food consumption along with greater hypertrophy of key organs for O2 transport.

Origin of chemiluminescence accompanying the reaction of the 9-cyano-10-methylacridinium cation with hydrogen peroxide.

The 9-cyano-10-methylacridinium cation possesses an electrophilic center at the carbon atom in position (9) susceptible to the addition of anions. The addition of OOH(-) to this cation--in weakly acidic, neutral, or alkaline media--initiates processes leading to the formation of electronically excited 10-methyl-9-acridinone, which deactivates by light emission. The effect of changes in reactant concentrations and pH on emission decay with time, as well as other features of the accompanying chemiluminescence, were established. Calculations carried out at the semiempirical and density functional theory level demonstrated that initial addition of OOH(-) and subsequent processes lead either to the elimination of OCNH (in weakly acidic and neutral media) or OCN(-) (in alkaline media) and that their exothermicity is sufficiently high to generate electronically excited 10-methyl-9-acridinone. On the other hand, primary addition of OH(-) to C(9) in alkaline media initiates the conversion of the cation to the nonexcited 10-methyl-9-acridinone. This relatively rapid process influences to a substantial extent the intensity of the chemiluminescence. The prospects for the analytical application of 9-cyano-10-methylacridinium salts are briefly outlined.

Spectral and acid-base features of 3,7-dihydroxy-2,8-diphenyl-4H,6H-pyrano[3,2-g]chromene-4,6-dione (diflavonol)--a potential probe for monitoring the properties of liquid phases.

Diflavonol is a molecule that can exist in neutral or anionic form and in several tautomeric forms in ground and excited states. Absorption and emission spectroscopy combined with theoretical calculations have shown that only one tautomer of neutral diflavonol exists in the ground state, but two exist in the excited state. In the latter case, one is the tautomer originating from the ground state tautomer, which exists in strongly protic solvents, the other is the phototautomer occurring in weakly protic or aprotic solvents as a result of the intramolecular transfer of one proton. The OH groups present in diflavonol and involved in weak intramolecular hydrogen bonds exhibit a proton-donating ability reflected by the experimental values of acidity constants or theoretical enthalpies and free energies of proton detachment. The electronically excited molecule is a relatively strong acid when it loses one proton. With increasing basicity of the medium, monoanionic and dianionic forms occur which exhibit spectral characteristics and an emission ability different from those of neutral diflavonol. These interesting features of diflavonol open up possibilities for the analytical use of the compound and its application as a spectral probe sensitive to the properties of liquid phases.

Spatial genetic structure and clonal diversity of island populations of lady's slipper (Cypripedium calceolus) from the Biebrza National Park (northeast Poland).

Three populations of the rare and endangered plant species Cypripedium calceolus were included in a study of genetic diversity and spatial genetic structure in the Biebrza National Park, northeast Poland. Analysis of 11 allozyme loci indicate that the populations of this species contained high genetic variability (P = 45.5%, A= 1.73). On the other hand, the genetic differentiation (FST = 0.014, P < 0.05) among C. calceolus populations was very low when compared to other species with similar life history characteristics. The observed high rate of gene flow (Nm = 18) may suggest that the populations studied derived from each other in the recent past. Five polymorphic allozyme markers identified 109 multilocus genotypes in three populations and the majority of them (67%) were population-specific. One of the populations studied, characterized by particularly extensive vegetative reproduction, showed the lowest clonal diversity (G/N = 0.15) and heterozygosity (HO = 0.111) values and the highest FIS(0.380), when compared to other two populations (G/N = 0.26-0.27, HO= 0.166-178, FIS = 0.024-0.055). This may indicate that clonal reproduction has an important influence on the genetic structure of C. calceolus populations. The longevity of genets, the out-crossing breeding system and the presence of recruitment from seeds are factors maintaining genetic diversity in C. calceolus.

Further studies on cytostatic activity of alkoxymethyl purine and pyrimidine acyclonucleosides.

The influence of 14 acyclonucleosides, derivatives of adenine, guanine, uracil and thymine on the phosphorylation of dAdo, dGuo, dCyd and dThd occurring in the cytosol of growing amelanotic melanoma transplanted to Syrian hamsters, as well as on inhibition of tumor growth were studied. From among the studied ACNs eight were tested earlier (Modrzejewska et al., 1996, The influence of alkoxymethyl purine and pyrimidine acyclonucleosides on growth inhibition of Kirkman-Robbins hepatoma and possible mechanism of their cytostatic activity, Z. Naturforch. 51c, 75-80); from among the newly synthesized ACNs, 1,3-N,N-diallyloxymethylthymine (AMT2), 1-N-allyloxymethyl-5,6-tetramethyleneuracil (AMUTM), and tested previously 1-N-allyloxymethylthymine (AMT1), administered i.p. in a dose of 0.2 mmol/kg body weight reduce the tumor mass from 0.98 g to 0.64 g +/- 0.11 g (i.e. 35% +/- 12%). 48 hours after i.p. administration of the mentioned ACNs in the same dose a reduction of tumor mass is accompanied by the inhibition of dAMP, dGMP and dTMP synthesis. AMT1 inhibits dThd phosphorylation from 6.2 to 4.22; AMT2 suppresses dAdo, dGuo and dThd phosphorylation by, correspondingly, from 2.8 to 1.7, from 10.8 to 7.5 and from 6.2 to 4.2; AMUTM depresses dAMP synthesis from 2.8 to 1.6 (all data: mumol of 2'dNMP formed per mg of protein per min. x 10(-4)). None of the 14 studied acyclonucleosides influences dCMP synthesis. In vivo, after hydration of allyloxymethyl group to hydroxypropoxymethyl residue (having -CH2OH group), AMT1, AMT2 and AMUTM undergo phosphorylation to corresponding triphosphates. Phosphorylated ACNs are not incorporated into tumor DNA, however they inhibit dAdo, dGuo and dThd incorporation into DNA. It is concluded that ACN triphosphates are not substrates for DNA polymerase but, competing with dATP dGTP and dTTP, inhibit incorporation of these 2'dNTP into DNA and, in consequence, reduce tumor growth, which is presumed to be the main mechanism of cytostatic activity of the studied ACNs.

Androgenic--anabolic steroids and body dysmorphia in young men.

There has recently been increasing attention focused on the use of androgenic-anabolic steroids (AAS). Some research has suggested that a disturbance in body image in males leads to AAS use. At present, the available literature is sparse and there is little/no discussion on the causal factors for their use. This review gives a historical account of the development and the changing patterns of AAS use. Both the physical and psychological side-effects are presented. The available evidence and theories for external/social and internal, psychological, and developmental influences are discussed in relation to AAS use. The current trends for the treatment of AAS abusers are also presented. Recommendations regarding further research are made in the conclusions.

Activity of exoglycosidases of the rat abdominal organs in untreated hemorrhagic shock.

Exoglycosidases catabolize glycoconjugates (proteoglycans, glycolipids and glycoproteins) at a rate depending on tissue and pathological changes. We determined exoglycosidase activity in successive sections of alimentary tract, spleen, liver and kidney of rats subjected to hemorrhagic shock. We found significant difference in exoglycosidase activity between alimentary tract segments in control animals and no significant changes in homologous tissues of the shocked rats comparing with control.

Acyclonucleoside analogues consisting of 5- and 5,6-substituted uracils and different acyclic chains: inhibitory properties vs purified E. coli uridine phosphorylase.

Synthetic procedures are described for the preparation of a variety of pyrimidine acyclonucleoside analogues, in which the aglycones are 5- and 5,6-substituted uracils, and the ribose moiety is replaced by different acyclic chains. These were examined as potential inhibitors of purified E. coli uridine phosphorylase. None of the compounds was a substrate for uridine phosphorylase, or either a substrate or inhibitor of E. coli thymidine phosphorylase. Kinetic measurements were employed to determine inhibition constants, Ki, for inhibition of uridine phosphorylase. One of the more effective of these was 1-(1',3'-dihydroxy-2'-propoxy)methyl-5,6-tetramethyleneuracil, with Ki = 2.7 microM. The same compound was a reasonably good inhibitor of the reverse, synthetic, reaction, with Ki values of 19 microM vs uracil as the variable substrate, and 15 microM vs alpha-D-ribose-1-phosphate as the variable substrate. For one of the analogues, which was a racemate, 1-(2',3'-dihydroxypropyl)-5,6-tetramethyleneuracil, it was shown that only one of the enantiomers (R) was an inhibitor, the (S) enantiomer being totally inactive. For several of the analogues, the corresponding isomeric N(3)-acyclonucleosides were inactive as inhibitors. The results for several of the good inhibitors were compared with those of other observers for inhibition of uridine phosphorylase from mammalian sources. Preliminary measurements with several of our analogues demonstrated that some of them were indeed one to two orders of magnitude more effective against the enzyme from mammalian sources.