Peripheral glutamate and TNF-α levels in patients with intracerebral hemorrhage: Their prognostic values and interactions toward the formation of the edemal volume

Objective We aimed to evaluate the prognostic values, contribution and interactions of the peripheral blood plasma glutamate and tumor-necrosis factor-α (TNF-α) levels toward the formation of the perifocal edema in patients with intracerebral hemorrhage (ICH). Methods Fifty patients with ICH and fifty healthy controls were included in the study. The peripheral markers were detected by high-sensitivity ELISA. Results A highly significant differences in plasma glutamate and TNF-α levels with good separation of their values was detected between patients and healthy controls. The two variables correlated with the severity of the symptoms and the initial volume of the ICH at admission. Both peripheral glutamate and TNF-α levels at admission were estimated as significant predictors for the formation of the perifocal edema five days after ICH; nevertheless, it was shown that they independently contribute to the development of the edema, without effects of interaction and regardless the localization of the ICH. Conclusions Our results support the idea for the significance of glutamate and TNF-α as peripheral markers for excitotoxicity and inflammation in ICH patients. The developed multiple regression model for prediction of the development of the edema could be beneficial in decision making between conservative treatment and surgical intervention in the clinical practice

Validation of the ELISA Method for Quantitative Detection of TNF-α in Patients with Intracerebral Hemorrhage

AIM: We aimed to investigate the sensitivity, reproducibility and validity of the commercial ELISA kits for quantitative detection of TNF-α and their potential application for screening purposes in patients with ICH.METHODS: Analysis of six independent standard series, evaluation of the deviation of the TNF-α concentration in patients with ICH, standard addition and visual analysis of whole UV-Vis spectra were carefully performed.RESULTS: Low standard deviations of the absorbance were detected for every standard, as well as in the samples of healthy controls and patients with ICH. The standard addition series have also confirmed high sensitivity and reproducibility of the assay, with a congruent shift of the standard curves with the concentration of TNF-α for the added plasma. The visual analyses of the gained spectra have revealed the absence of any matrix effects from the addition of the human plasma in the reconstituted standards.CONCLUSION: The commercial ELISA kits can be used in the clinical practice for screening purposes of the plasma TNF-α levels in patients with ICH

Small animal model in the development of radiopharmaceuticals - the step forward to clinical studies

Experimental design is obviously a critical component for the success of any research activities concerning development and evaluation of new radiopharmaceuticals. The development of new generation of radiolabeled molecules and technologies (SPECT/PET) for delivery of radioisotopes to the disease-target-sites with a high degree of precision, recognition, and target selectivity can facilitate mapping of the biochemistry of the metabolic organ function, visualizing the molecular biology of cell function, and zooming in on gene function for delineating differences in molecular biology of normal health from disease, in animals to humans. This type of imaging refines diagnostic differentiation at molecular or metabolic levels between health and disease, and among various diseases, often leading to more effective therapy. Experimental animal models have substantially contributed to a better understanding of mechanisms of disease and show the novel approaches in imaging and image analysis were equally important to meet the challenges of analyzing the complex mechanisms underlying pathophysiological processes in vivo. The appropriate animal models are not only helpful in finally proving longstanding hypotheses for development for radiopharmaceuticals but have rather been used to discover new and previously unexpected mechanisms and causal relationships. Proper animal models are key factor in successful pharmaceutical or medicinal experiment. To reduce animal number for ethical and financial reasons, cost-efficient methods where high quantities of data are achieved fast are optimal. Biodistribution and pharmacokinetics studies diagnostic or therapeutic radiopharmaceuticals by SPECT or PET imaging followed by post mortem analysis in diseases model gives start point for further steps toward clinical applications. In this presentation, targeting properties, biodistribution and pharmacokinetics of different molecules, as potential radiopharmaceuticals have been studied in small animal models using suitable imaging modalities and post mortem analysis. The following experimentally designed animal models have been introduced in our work so far, with the intention of showing the importance of establishing procedures and protocols for experimental studies as an essential part in the development of new radiopharmaceutical products and quality control of existing radiopharmaceutical products. The rats were used for establishing of: - stasis-induced thrombus in the femoral vein after injection of thromboplastin to demonstrate Deep Venous Thrombosis using radiolabeled Tirofiban - GPIIb/IIIa inhibitor; - induced amyloidosis by multiple application of beta2-microglobulin for determination of the existence of the depositing osteoarticular tissues, condition associated with hemodialysis in patients with chronic kidney diseases - collagen-induced arthritis as a model of inflammatory arthritis - bacterial abscesses by the injection of Staphylococcus aureus Mice animal models were used for: - in vivo evaluation of the radiolabelled conjugated antibodies in normal Balb/c mice and nude mice xenografts - per os administration of iodine labeled BSA loaded microspheres to show the strong adjuvant effect by inducing IgA secretion at the genito-urinary mucosa - athymic nude mice tumor bearning to demonstrate specifity of pretargeting technique referred to the Affinity Enhancement System (AES) uses bispecific antibodies and radiolabeled bivalent haptens; The use of experimental animal models in the design of new drugs including radiopharmaceuticals is a key part of preclinical trials. Usually this approach can never replicate human disease or the varied and complex physical and psychological manifestations of human conditions. For these reasons the process of experimental design should be carried out routinely to ensure the generation of valid, reproducible and published data

Temperature Effects Explain Continental Scale Distribution of Cyanobacterial Toxins

Insight into how environmental change determines the production and distribution of cyanobacterial toxins is necessary for risk assessment. Management guidelines currently focus on hepatotoxins (microcystins). Increasing attention is given to other classes, such as neurotoxins (e.g., anatoxin-a) and cytotoxins (e.g., cylindrospermopsin) due to their potency. Most studies examine the relationship between individual toxin variants and environmental factors, such as nutrients, temperature and light. In summer 2015, we collected samples across Europe to investigate the effect of nutrient and temperature gradients on the variability of toxin production at a continental scale. Direct and indirect effects of temperature were the main drivers of the spatial distribution in the toxins produced by the cyanobacterial community, the toxin concentrations and toxin quota. Generalized linear models showed that a Toxin Diversity Index (TDI) increased with latitude, while it decreased with water stability. Increases in TDI were explained through a significant increase in toxin variants such as MC-YR, anatoxin and cylindrospermopsin, accompanied by a decreasing presence of MC-LR. While global warming continues, the direct and indirect effects of increased lake temperatures will drive changes in the distribution of cyanobacterial toxins in Europe, potentially promoting selection of a few highly toxic species or strains.Peer reviewe

Dominance of Microcystis spp. in Lake Dojran – a consequence of 30 years of accelerated eutrophication

In the past 30 years, Lake Dojran (Macedonia) has experienced extreme periods of droughts and water removal for irrigation purposes that have resulted in an accelerated process of eutrophication leading to hypertrophy. In the present paper, we document successive changes of the microflora in plankton communities as a result of the prolonged eutrophication impact over this period. The original well established phytoplankton community in the lake, which was rich in species (especially coccoid green forms belonging to the genera Pediastrum, Scenedesmus, Staurastrum and Tetraedon) and which followed the natural annual succession in the form of diatoms > green algae > cyanobacteria, has been greatly modified over the years. Two episodes of intensive ‘water blooms’ have been recorded, one involving Gloeotrichia natans (in 1993/94) and the other the dinoflagellates Ceratium hirudinella and C. monoceros (1996/97), events that indicated drastic changes of the ecological conditions. Dominance of Dolichospermum aff. flosaquae was recorded in 2008. Finally, the phytoplankton in Lake Dojran completed a turnover towards the overall dominance of Microcystis spp., with a total of nine co-existing species. The first documented record of Microcystis protocystis outside pan- and neotropical regions is here presented. The described successions in phytoplankton species composition and dominance are attributed to gradual and constant changes in the nutrient status of the lake in the direction of hypertrophy, the basic nutritive parameters, total P and N content, having increased more than 40- and eight-fold, respectively, over the years. Measurements of cyanobacterial toxicity have been few, and they were sporadically performed. Nonetheless, the presence of microcystins in the water has been documented, with indications of significant concentration peaks. The paper is intended to be a tribute to all scientific workers who, like Professors Jelena and Živojin Blaženčić, have devoted their expertise and time to the task of revealing forced changes in the lake’s biota in the hope of creating a favourable atmosphere for immediate human intervention to save this once very rich and diverse ecosystem

Surgical Outcome in Patients with Spontaneous Supratentorial Intracerebral Hemorrhage

The aim of the paper was to evaluate the surgical outcome in patients with spontaneous supratentorial intracerebral hemorrhage (ICH) after surgical intervention, in respect to the initial clinical conditions, age, sex, hemispheric side and anatomic localization of ICH. Thirty-eight surgically treated patients with spontaneous supratentorial intracerebral hemorrhage were included in the study. The surgical outcome was evaluated three months after the initial admission, according to the Glasgow Outcome Scale (GOS). The surgical treatment was successful in 14 patients (37%), whereas it was unsuccessful in 24 patients (63%). We have detected a significant negative correlation between the Glasgow Coma Scale (GCS) scores on admission and the GOS scores after three months, suggesting worse neurological outcome in patients with initially lower GCS scores. The surgical outcome in patients with ICH was not affected by the sex, the hemispheric side and the anatomic localization of ICH, but the age of the patients was estimated as a significant factor for their functional outcome, with younger patients being more likely to be treated successfully. The surgical outcome is affected from the initial clinical state of the patients and their age. The treatment of ICH is still an unsolved clinical problem and the development of new surgical techniques with larger efficiency in the evacuation of the hematoma is necessary, thus making a minimal damage to the normal brain tissue, as well as decreasing the possibility of postoperative bleeding

Stratification strength and light climate explain variation in chlorophyll a at the continental scale in a European multilake survey in a heatwave summer

To determine the drivers of phytoplankton biomass, we collected standardized morphometric, physical, and biological data in 230 lakes across the Mediterranean, Continental, and Boreal climatic zones of the European continent. Multilinear regression models tested on this snapshot of mostly eutrophic lakes (median total phosphorus [TP] = 0.06 and total nitrogen [TN] = 0.7 mg L-1), and its subsets (2 depth types and 3 climatic zones), show that light climate and stratification strength were the most significant explanatory variables for chlorophyll a (Chl a) variance. TN was a significant predictor for phytoplankton biomass for shallow and continental lakes, while TP never appeared as an explanatory variable, suggesting that under high TP, light, which partially controls stratification strength, becomes limiting for phytoplankton development. Mediterranean lakes were the warmest yet most weakly stratified and had significantly less Chl a than Boreal lakes, where the temperature anomaly from the long-term average, during a summer heatwave was the highest (+4 degrees C) and showed a significant, exponential relationship with stratification strength. This European survey represents a summer snapshot of phytoplankton biomass and its drivers, and lends support that light and stratification metrics, which are both affected by climate change, are better predictors for phytoplankton biomass in nutrient-rich lakes than nutrient concentrations and surface temperature.Peer reviewe