Aminoguanidine and N-acetyl-cysteine supress oxidative and nitrosative stress in EAE rat brains

Experimental autoimmune encephalomyelitis (EAE) is a well-established animal model of human multiple sclerosis (MS). We have evaluated the role of oxidative and nitrosative stress, as the causal factors in the development of EAE, responsible for the damage of cardinal cellular components, such as lipids, proteins and nucleic acids, resulting in demyelination, axonal damage, and neuronal death. EAE was induced in female Sprague-Dawley rats, 3 months old (300±20 g), by immunization with myelin basic protein in combination with Complete Freund's adjuvant (CFA). The animals were divided into seven groups: control, EAE, CFA, EAE+aminoguanidine (AG), AG, EAE+N-acetyl-L-cysteine (NAC) and NAC. The animals were sacrificed 15 days after EAE induction, and the levels of nitrosative and oxidative stress were determined in 10% homogenate of the whole encephalitic mass. In EAE rats, brain NO production and MDA level were significantly increased (P<0.001) compared to the control values, whereas AG and NAC treatment decreased both parameters in EAE rats compared to EAE group (P<0.001). Glutathione (GSH) was reduced (P<0.001) in EAE rats in comparison with the control and CFA groups, but increased in EAE+AG and EAE+NAC group compared to the EAE group (P<0.01). Superoxide dismutase (SOD) activity was significantly decreased (P<0.001) in the EAE group compared to all other experimental groups. The clinical expression of EAE was significantly decreased (P<0.05) in the EAE groups treated with AG and NAC compared to EAE rats, during disease development. The obtained results prove an important role of oxidative and nitrosative stress in the pathogenesis of EAE, whereas AG and NAC protective effects offer new possibilities for a modified combined approach in MS therapy.     

Urinary Immunoglobulin G to Albumin Ratio and N-Acetyl-Beta-D-Glucosaminidase as Early Predictors of Therapeutic Response in ANCA-Associated Glomerulonephritis

Background

The aim of our study was to evaluate the prognostic value of glomerular and tubular proteinuria and tubular enzymuria as early indicators of therapeutic response to induction therapy with i.v. pulse cyclophosphamide (CyC) and methylprednisolone (MP) in patients with antineutrophil cytoplasmic antibody (ANCA) associated glomerulonephritis.

Methods and Findings

An observational single-center study was conducted in 30 patients with ANCA-associated glomerulonephritis. Patients were divided into subgroups with good or poor response to CyC therapy according to clinical and laboratory parameters. The diagnosis of ANCA-associated glomerulonephritis was based on the Chapel-Hill disease definitions. Good response to induction therapy was significantly associated with higher absolute values of urine N-acetyl-beta-D-glucosaminidase (NAG) to creatinine ratio (above 14.83 microcat/mol) and urine immunoglobulin G (IgG) to albumin ratio (above 0.09) at the time of diagnosis, while albuminuria or proteinuria did not have any early predictive value. The remission of renal disease was anticipated as early as 3 months after introduction of induction therapy in patients with reduction of urine NAG to creatinine ratio below the baseline value and in patients with at least 24% rise in eGFR.

Conclusions

Urine IgG to albumin and urine NAG to creatinine ratio are better early predictors of treatment response in patients with ANCA-associated glomerulonephritis than proteinuria or albuminuria.     

Serotonin localization and its functional significance during mouse preimplantation embryo development

Serotonin is a neurotransmitter functioning also as a hormone and growth factor. To further investigate the biological role of serotonin during embryo development, we analysed serotonin localization as well as the expression of specific serotonin 5-HT1D receptor mRNA in mouse oocytes and preimplantation embryos. The functional significance of serotonin during the preimplantation period was examined by studying the effects of serotonin on mouse embryo development. Embryo exposure to serotonin (1 microM) highly significantly reduced the mean cell number, whereas lower concentrations of serotonin (0.1 microM and 0.01 microM) had no significant effects on embryo cell numbers. In all serotonin-treated groups a significant increase in the number of embryos with apoptotic and secondary necrotic nuclei was observed. Expression of serotonin 5-HT1D receptor mRNA in mouse oocytes and preimplantation embryos was confirmed by in situ hybridization showing a clearly distinct punctate signal. Immunocytochemistry results revealed the localization of serotonin in oocytes and embryos to the blastocyst stage as diffuse punctate cytoplasmic labelling. It appears that endogenous and/or exogenous serotonin in preimplantation embryos could be involved in complex autocrine/paracrine regulations of embryo development and embryo-maternal interactions.     

Inhibition of α-glucosidase activity by curcumin loaded on ZnO@rGO nanocarrier for potential treatment of diabetes mellitus

Curcumin (Cur) is an acidic polyphenol with some effects on α-glucosidase (α-Glu), but Cur has disadvantages such as being a weak target, lacking passing the blood–brain barrier and having low bioavailability. To enhance the curative effect of Cur, the hybrid composed of ZnO nanoparticles decorated on rGO was used to load Cur (ZnO@rGO-Cur). The use of the multispectral method and enzyme inhibition kinetics analysis certify the inhibitory effect and interaction mechanism of ZnO@rGO-Cur with α-Glu. The static quenching of α-Glu with both Cur and ZnO@rGO-Cur is primarily driven by hydrogen bond and van der Waals interactions. The conformation-changing ability by binding to the neighbouring phenolic hydroxyl group of Cur increased their ability to alter the secondary structure of α-Glu, resulting in the inhibition of enzyme activity. The inhibition constant (K_{i, Cur} > K_{is,ZnO@rGO-Cur} ) showed that the inhibition effect of ZnO@rGO-Cur on α-Glu was larger than that of Cur. The CCK-8 experiments proved that ZnO@rGO nanocomposites have good biocompatibility. These results suggest that the therapeutic potential of ZnO@rGO-Cur composite is an emerging nanocarrier platform for drug delivery systems for the potential treatment of diabetes mellitus.     

Orientation of the pit-building antlion larva Euroleon (Neuroptera, Myrmeleontidae) to the direction of substrate vibrations caused by prey

Pit-building antlion Euroleon nostras constructs efficient traps in sand to catch its prey. The predator is known to react to substrate vibrations produced by movements of its prey outside the pit with sand-tossing behaviour but it has not yet been ascertained if this reaction is directed towards the prey. The accuracy of the sand-tossing response in the presence of four prey species was measured using a video recording method. The sand-tossing angle was highly positively correlated with the prey angle. Sand tossing was most frequently elicited when prey was on the posterior sand surface. Covering the larval photoreceptors did not influence the antlion's localizing behaviour.     

Validation of an individualised model of human thermoregulation for predicting responses to cold air

Most computer models of human thermoregulation are population based. Here, we individualised the Fiala model [Fiala et al. (2001) Int J Biometeorol 45:143–159] with respect to anthropometrics, body fat, and metabolic rate. The predictions of the adapted multisegmental thermoregulatory model were compared with measured skin temperatures of individuals. Data from two experiments, in which reclining subjects were suddenly exposed to mild to moderate cold environmental conditions, were used to study the effect on dynamic skin temperature responses. Body fat was measured by the three-compartment method combining underwater weighing and deuterium dilution. Metabolic rate was determined by indirect calorimetry. In experiment 1, the bias (mean difference) between predicted and measured mean skin temperature decreased from 1.8°C to −0.15°C during cold exposure. The standard deviation of the mean difference remained of the same magnitude (from 0.7°C to 0.9°C). In experiment 2 the bias of the skin temperature changed from 2.0±1.09°C using the standard model to 1.3±0.93°C using individual characteristics in the model. The inclusion of individual characteristics thus improved the predictions for an individual and led to a significantly smaller systematic error. However, a large part of the discrepancies in individual response to cold remained unexplained. Possible further improvements to the model accomplished by inclusion of more subject characteristics (i.e. body fat distribution, body shape) and model refinements on the level of (skin) blood perfusion, and control functions, are discussed.     

The mechanism of amyloid-fibril formation by stefin B: temperature and protein concentration dependence of the rates

Cystatins, a family of structurally related cysteine proteinase inhibitors, have proved to be useful model system to study amyloidogenesis. We have extended previous studies of the kinetics of amyloid-fibril formation by human stefin B (cystatin B) and some of its mutants, and proposed an improved model for the reaction. Overall, the observed kinetics follow the nucleation and growth behavior observed for many other amyloidogenic proteins. The minimal kinetic scheme that best fits measurements of changes in CD and thioflavin T fluorescence as a function of protein concentration and temperature includes nucleation (modeled as N(I) irreversible transitions with equivalent rates (k(I)), which fitted with N(I) = 64), fibril growth and nonproductive oligomerization, best explained by an off-pathway state with a rate-limiting escape rate. Three energies of activation were derived from global fitting to the minimal kinetic scheme, and independently through the fitting of the individual component rates. Nucleation was found to be a first-order process within an oligomeric species with an enthalpy of activation of 55 +/- 4 kcal mol(-1). Fibril growth was a second-order process with an enthalpy of activation (27 +/- 5 kcal mol(-1)), which is indistinguishable from that of tetramer formation by cystatins, which involves limited conformational changes including proline trans to cis isomerization. The highest enthalpy of activation (95 +/- 5 kcal mol(-1) at 35 degrees C), characteristic of a substantial degree of unfolding as observed prior to domain-swapping reactions, equated with the escape rate of the off-pathway oligomeric state.     

Density-dependent individual growth of marble trout (<Emphasis Type="Italic">Salmo marmoratus</Emphasis>) in the Soca and Idrijca river basins,Slovenia

The aim of this field study was to investigate effects of estimated fish- and sea urchin herbivory on the reproductive potential of four species of macroalgae; Halimeda macroloba (Decasine), H. renschii (Hauck), Turbinaria ornata (Turner) and Padina boergesenii (Allender et Kraft). Fish and sea urchin herbivory were calculated based on reported consumption rates for their biomass estimates. We hypothesized that reduced herbivory would increase algal size and the reproductive potential, which may promote algal recruitment and be one of the driving mechanisms behind algal shifts and persistent algae-dominated reefs. Algae were investigated in field sites where the estimated fish- and or sea urchin herbivory differed. Our results suggest that algal fecundity of T. ornata and P. boergesenii are positively correlated to their size. Fecundity of T. ornata was higher and individuals grew larger in areas where estimated fish herbivory was lower. The two species of Halimeda grew larger and had higher fecundity in areas where estimated sea urchin herbivory was lower. P. boergesenii responded ambiguously to patterns in herbivory. Due to species-specific responses to different herbivores, it is difficult to generalize about effects of overfishing on algal fecundity. Handling editor: S. Wellekens     

Synthesis and biological evaluation of some 17-picolyl and 17-picolinylidene androst-5-ene derivatives

Starting from dehydroepiandrosterone (1) 17-picolyl (2), 17-picolinylidene (7), 17-picolinylidene-16-one (10 and 11), and 17-picolyl-16-one (15) derivatives of androst-5-ene were synthesized in one, two, four and five steps respectively. By the Oppenauer oxidation or dehydration of 2, 7, 10, and 11 with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), the corresponding A and B ring modified derivatives 3, 5, 6, 8, 9, and 12-14 were obtained. The structure of 2 was unambiguously proved by the appropriate X-ray structural analysis. Compounds 3, 5, 9, 12-14 showed inhibitory activity against the enzyme aromatase. Antibacterial activity, toxicity to brine shrimp Artemia salina, antitumor activity against three tumor cell lines (human cervix carcinoma HeLa cells, human melanoma FemX cells, and human myelogenous leukemia K562 cells) and toxicity against peripheral blood mononuclear cells were evaluated. Three tested compounds, namely 11, 13, and 15, showed strong activity against all three cell lines, the IC(50) values being in the range of 4-10 microM.