Enhancement of benzodiazepine binding sites following chronic treatment with flumazenil

The aim of this study was to improve our knowledge of the mechanisms leading to adaptive changes in gamma-aminobutyric acid(A) (GABA(A)) receptors following chronic drug treatment. Exposure (48 h) of human embryonic kidney (HEK 293) cells stably expressing recombinant alpha1beta2gamma2S GABA(A) receptors to the antagonist of benzodiazepine binding sites, flumazenil (5 microM), enhanced the maximum number (B(max)) and the equilibrium dissociation constant (K(d)) of [3H]flunitrazepam binding sites. The flumazenil-induced enhancement in B(max) was potentiated by GABA (50 microM) and reduced by the GABA(A) receptor antagonist, bicuculline (100 microM). Flumazenil-induced enhancement in K(d) was affected by neither of these treatments. GABA (50 microM) enhanced the density of [3H]flunitrazepam binding sites, and this enhancement was greater in the presence of diazepam (1 microM). The results suggest that chronic flumazenil treatment up-regulates in a bicuculline-sensitive manner benzodiazepine binding sites at stably expressed GABA(A) receptors.     

Novel cyano- and N-isopropylamidino-substituted derivatives of benzo[b]thiophene-2-carboxanilides and benzo[b]thieno[2,3-c]quinolones: synthesis, photochemical synthesis, crystal structure determination, and antitumor evaluation. 2

Derivatives of 3-chlorobenzo[b]thiophene-2-carboxanilides and their "cyclic" analogues benzo[b]thieno[2,3-c]quinolones were synthesized. Spectroscopic study of the interactions of some representatives of "cyclic" derivatives and their "acyclic" precursors with ds-DNA/RNA supported strong intercalative binding of the former and weak nonintercalative binding of the latter group of compounds. All tested compounds showed a certain antiproliferative effect on a series of human tumor cells and on a normal cell line. Among the compounds, those with one amidino-substituent have shown the best effect. The most active benzo[b]thieno[2,3-c]quinolones induced apparent S and G2/M arrests of the cell cycle, which resulted in apoptosis. These results strongly suggest that the compounds may act as topoisimerase "poisons", which is in good agreement with their intercalative mode of binding to ds-DNA/RNA, in contrast to the studied "acyclic"group of derivatives. 6a and 6d showed the best selectivity by inhibiting the growth of tumor cells but not of normal fibroblasts.     

Relationship between the state of the surface of four commercial quartz flours and their biological activity in vitro and in vivo

Four commercial quartz dusts (flours), two inflammogenic in vivo and activating macrophages in vitro (Qz 2/1-c and Qz 3/1-c) and two mostly inert (Qz 5/1-c and Qz 11/1-c), have been compared regarding their surface properties, in order to detect chemical differences which may account for their different biological behaviour. The following features have been examined: 1) extent of the amorphous fraction (heat associated alpha<-->beta transition of quartz) and its solubility in HF; 2) potential to cleave a carbon-hydrogen bond with consequent generation of carbon centred radicals (spin trapping technique, EPR); 3) evolution of surface functionalities upon heating (FTIR spectroscopy); 4) mechanisms of adsorption of water on dusts outgassed at 150 degrees and at 800 degrees C (adsorption calorimetry). HCl treated samples have also been examined. The two "less toxic" quartzes are more resistant to HF attack, coordinate irreversibly H2O molecules and exhibit strong adsorption sites, which are absent in the other two and in a very pure quartz dust. Conversely all samples show the same potential to release free radicals. The different behaviour of the two sets of dust is consistent with a different level of impurities, namely aluminium ex kaolin, carbon and alkaline ions. The less inflammogenic quartzes appear to be covered by aluminium ions (and possibly iron) which strongly holds molecular water or carbonates, thus reducing the silanol patches to a large extent and changing the surface properties of the particles. We hypothesize that cellular response, and particularly macrophage activation and death, is mediated by strong interactions between silanol patches and some cell membrane components, but inhibited when the surface of the particle is modified by the presence of aluminium ions, surface carbonates and other metal contaminants. This hypothesis suggests that grinding procedures with little appropriate additives, e.g. kaolin, alumina, can reduce the biological activity of quartz dusts.     

An Eco-Epidemiological Study of Contamination of Soil with Infective Forms of Intestinal Parasites

The objectives of the present work were to screen topsoil samples collected from public squares in two cities within the Argentine Patagonia for the presence of infective forms of intestinal parasites and to examine the possible relationship between positive findings and the environmental, socioeconomic, and cultural conditions of that region. For this purpose we studied 13 public squares, their 13 custodians, and 44 family groups within their respective surrounding areas. Of the 226 topsoil samples analyzed, 44.3% proved positive for infective forms of intestinal parasites, with 17.3% of these containing more than one species. The frequency of appearance of positive samples was dependent on the season of the year (p < 0.001), while presence of the parasites was related to the soil pH (p < 0.05) but independent of the soil relative humidity (p > 0.05). Some of the organisms detected are associated with zoonoses. We observed the presence of Capillaria spp. and Spirocerca spp. under cool desert climatic conditions. Within the group of custodians we detected hematologic alterations one positive serology for toxoplasmosis and documented behavior conducive to risk of infection with the parasites found in those squares. Within the family group an acquaintance with parasitic zoonoses and their prevention was an inconsistent finding, with toxocarosis and toxoplasmosis being the diseases associated with the greatest degree of ignorance. Furthermore, we consider the failure to de-parasitize pets and the practice of feeding them with raw meat, as typically found in our family survey, to be factors contributing to a greater likelihood of public square contamination. From the results obtained here, we propose a spatial organization approach for the purpose of detecting zones at risk of contracting zoonotic parasitoses within urban environments.     

Bullying in Adolescent Residential Care: The Influence of the Physical and Social Residential Care Environment

Background

To date, no study examined possible contributions of environmental factors to bullying and victimization in adolescent residential care facilities.

Objective

By testing one part of the Multifactor Model of Bullying in Secure Setting (MMBSS; Ireland in Int J Adolesc Med Health 24(1):63–68, 2012), this research examined the way the physical and social residential environment relates to bullying and victimization in adolescent residential care.

Method

Young people aged 11–21 (N = 272) from ten residential institutions in Croatia completed: (a) an anonymous self-reported bullying questionnaire; (b) the social residential environment questionnaire; and (c) the physical residential environment questionnaire.

Results

The results demonstrated that both bullies and victims reported having significantly lower levels of perceived peer support than other residents. Male bullies also reported significantly lower levels of their overall wellbeing within their facilities and were significantly more likely than non-bullies to perceive their facilities as having problems with cleanliness and food. Male victims were significantly younger than non-victims. Female victims reported lower levels of their overall wellbeing than non-victims as well as poorer relationship with staff.

Conclusion

The results are discussed with reference to the relevant prison and school-based bullying literature and directions for future research are provided. Overall, the findings of this study are consistent with the part of the MMBSS (Ireland 2012) examined and provide initial support for the notion that the special nature of the physical and social residential environment may be important in explaining bullying in care.

     

ADMA and C-reactive protein as mortality predictors in dialysis patients

There is a higher mortality between patients with end-stage renal disease than patients in the general population. These circumstances have led to a search for risk factors as predictors of mortality in dialysis patients. Amongst those, inhibitors of the nitric-oxide (NO) synthesis deserve special attention, since patients with end-stage renal disease are also characterized by accelerated atherosclerosis. Asymmetric-dimethylarginine (ADMA) and symmetric-dimethylarginine (SDMA), as well as C-reactive protein (CRP), have also been recognized as predictors of mortality in patients on dialysis. The aim of our study was to compare the prediction power of ADMA, SDMA and CRP for all-cause mortality in patients with end stage renal disease during the fourteen month follow-up. In total 162 patients on hemodialysis were included. ADMA and SDMA were measured by the high-performance liquid chromatography (HPLC); CRP was measured using immunonephelometric assays. During the 14-month period 28 patients (34.1%) died from all-cause mortality. Using univariate analysis, hazard ratios (HR) of the potential independent predictors of mortality in hemodialysis patients were ADMA (HR 1.39 (1.01–1.91) p=0.043) and CRP (HR 1.024 (1.009–1.1.040) p=0.001). Further, multivariate analysis (MVA), however, showed that ADMA is the only predictor of all-cause mortality (HR 1.76 (1.002–3.11) P=0.049), while SDMA failed to predict death in this population. Therefore, our data shows that ADMA is an independent and better marker of all-cause mortality compared with CRP.     

The effect of ribavirin on reactive astrogliosis in experimental autoimmune encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is an animal model of CNS inflammatory and demyelinating disease multiple sclerosis. Microglia and astrocytes represent two related cell types involved in the brain pathology in EAE. Accumulations of hypertrophic reactive astrocytes, intensely stained with glial fibrillary acidic protein (GFAP), which also expressed vimentin, are prominent features of EAE lesions. Recent studies from our laboratory reported that ribavirin attenuated the disease process in EAE by reducing clinical and histological manifestations. EAE was induced in genetically susceptible Dark Agouti rats with syngeneic spinal cord homogenate in complete Freund's adjuvant. Real time PCR and immunohistochemistry were used for determination of GFAP and vimentin gene and tissue expression. We have observed the increased gene and tissue expression of GFAP and vimentin in EAE rats. Ribavirin treatment significantly decreased the number of reactive astrocytes at the peak of disease. At the end of the disease, we have observed reactive GFAP(+) and vimentin(+) astrocytes in both immunized and ribavirin-treated groups, accompanied by increased level of GFAP mRNA. The present study indicates that ribavirin may have the ability to attenuate astrocyte proliferation and glial scaring at the peak of the disease and modulate the astroglial response to EAE during the time-course of the disease.     

Transepithelial transport of biperiden hydrochloride in Caco-2 cell monolayers

The aim of this research has been to determine the biperiden hydrochloride permeability in Caco-2 model, in order to classify it based on the Biopharmaceutics Classification System (BCS). The World Health Organization (WHO) as well as many other authors have provisionally assigned the drug as BCS class I (high solubility-high permeability) or III (high solubility-low permeability), based on different methods. We determined biperiden BCS class by comparing its permeability to 5 pre-defined compounds: atenolol and ranitidine hydrochloride (low permeability group) and metoprolol tartrate, sodium naproxen and theophylline (high permeability group). Since biperiden permeability was higher than those obtained for high permeability drugs, we classified it as a BCS class I compound. On the other hand, as no differences were obtained for permeability values when apical to basolateral and basolateral to apical fluxes were studied, this drug cannot act as a substrate of efflux transporters. As a consequence of our results, we suggest that the widely used antiparkinsonian drug, biperiden, should be candidate for a waiver of in vivo bioequivalence studies.