Studies on forest health and vegetation changes in Greece under the effects of climate changes

Greece, as part of the Mediterranean Basin, is projected to be among the most vulnerable countries to climate change. It is therefore quite urgent to adapt forest management to the changing climate in order to enhance biodiversity and to enable the conservation of healthy and productive forests. In the framework of the project LIFE+ AdaptFor (www.life-adaptfor.gr), an effort was made to understand the ecological responses and the vulnerability of forest ecosystems in the face of climate change; the overall aim of the project is the development and implementation of appropriate adaptation strategies. Four study areas were selected where changes in vegetation, quite likely attributed to climate change, have already been observed (dieback of Scots pine and Greek fir, intrusion of conifers in broadleaved forests). To investigate the synergism of climatic parameters’ alterations in the development of the occurring phenomena, time series of temperature and precipitation for the period 1950–2009 were produced and parameters of forest status were investigated, including mapping of vegetation changes through remote sensing. The findings support the hypothesis that climate change has an impact on forest health; the dieback of tree species can be attributed to outbreaks of pathogens (fungi and insects) which are associated with climatic parameters. However, the intrusion of conifer species into broadleaved forests showed no direct connection to climatic parameters, something that needs to be further investigated. In all cases, insufficient or inappropriate management practices applied in the areas exacerbated the occurring phenomena.     

Pituitary adenylate cyclase-activating peptide: a pivotal modulator of steroid-induced reproductive behavior in female rodents

Pituitary adenylate cyclase-activating polypeptide (PACAP) regulates the secretion of GnRH into the hypothalamic hypophysial portal system and sensitizes the pituitary for release of hormones that trigger ovulation. Because reproductive behavior is synchronized with GnRH release, the present study was undertaken to determine whether PACAP in the ventromedial nucleus (VMN) plays a role in receptivity. To this end, we used rat and mouse reproductive behavioral models to determine the biological relationship between PACAP and steroid receptor function in females. We provide evidence for the requirement of PACAP in the VMN for progesterone (P)-dependent sexual behavior in estrogen (E)-primed females. We clarify the biological and molecular mechanisms of PACAP activity by showing 1) that inhibition of endogenous PACAP suppresses P receptor (PR)-dependent sexual behavior facilitated by the steroid P or D1-like agonist SKF38393 and 2) that PR, steroid receptor coactivators-1 and -2, and new protein synthesis are essential for ligand independent PACAP-facilitated behavior. These findings are consistent with convergence of PACAP-mediated cellular signals on PR for genomic activation and subsequent behavioral changes. Further, we show that steroids regulate both endogenous PACAP mRNA in the VMN and immunoreactive PACAP in the medial basal hypothalamus and cerebral spinal fluid for ligand-dependent, steroid receptor-dependent receptivity. The present findings delineate a novel, steroid-dependent mechanism within the female hypothalamus by which the neuropeptide PACAP acts as a feed-forward, paracrine, and/or autocrine factor for synchronization of behavior coordinate with hypothalamic control of ovulation.     

In situ cooling in a lung hilar clamping model of ischemia-reperfusion injury

Experimental models for studying transplantation have up to now been unable to isolate reperfusion injury with minimal surgical manipulation and without the interference of graft rejection. Six pigs were subjected to left hilum preparation only (control group), and eight pigs were subjected to left hilum preparation plus in situ cooling ischemia and reperfusion of the lung (experimental group). The hilum was dissected free from other tissues in both groups. Lung preservation was achieved by antegrade flush perfusion via the left pulmonary artery. Pulmonary veins were clamped at the left atrium and a vent was created. The left main bronchus was clamped. Lung temperature was maintained at 4 degrees -8 degrees C, while core temperature was kept at 38 degrees C. After 3 hrs of cold ischemia the clamps were removed and the lung was reperfused. Elevated pulmonary vascular resistance and local and systemic aspects of ischemia-reperfusion syndrome were consistently reproduced. This large-animal model of in situ unilateral lung cold ischemia with warm reperfusion proved to be very reliable in reproducing all aspects of ischemia-reperfusion injury. It excludes the interference of rejection and extensive surgical manipulation. We therefore propose its use in experimental studies investigating pharmaceutical or cooling modifications affecting lung ischemia-reperfusion outcomes.     

Different intrafamilial clinical presentation of FMF mutation carriers

Familial Mediterranean fever (FMF) is a heterogeneous disorder; at present, it is diagnosed using only genetic methods. In the current study, we performed molecular analysis in two families presenting with FMF. In the first family, we report two brothers with a common genotype (M694V/V726A) but with different clinical presentation. In the second family, we identified the M694V and K695R mutations in a presymptomatic carrier.     

In vitro evaluation for potential calcification of biomaterials used for staple line reinforcement in lung surgery

Bovine pericardium (BPC) and polytetrafluoroethylene (PTFE) have been widely used to reinforce staple lines in lung resection. Since limited information regarding the calcification of these biomaterials is available, we undertook an in vitro study to evaluate their calcification potential. Commercially available BPC and PTFE biomaterials were evaluated and compared with custom-prepared BPC tissue. In vitro calcification was performed via submersion in supersaturated solution in a double-walled glass reactor at 37.0 degrees C +/- 0.1 degrees C, pH 7.4 +/- 0.1, mimicking most ion concentrations of human blood plasma. In processing of calcification, the pH decrease of the solution simulated the addition of consumed H(+), Ca(2+), and PO(4)(3-) ions from titrant solutions, the concentrations of which were based on the stoichiometry of octacalcium phosphate. The molar ion addition with time was recorded, and the initial slope of the curve was computed for each experiment. The rate of calcification developed (molar calcium phosphate ion addition rate per time and total surface area) (R) was computed after that with respect to the relative supersaturation (sigma) used in each experiment. R for custom-prepared BPC tissues was found to be in the range of 0.19 +/- 0.08 to 0.52 +/- 0.19 (n = 17) in sigma range of 0.72 to 1.42. Commercial BPC was found to be 0.016 to 0.052 (n = 4), and PTFE was 0.005 to 0.05 (n = 8) in the same sigma range. Both clinically applied biomaterials, BPC and PTFE, seemed to be calcified with rates of at least one order of magnitude lower than the custom-prepared BPC tissue. This data suggested that BPC and PTFE biomaterials showed a similar, relatively very low tendency for calcification compared with custom-prepared BPC tissue. Although further studies are necessary, staple line reinforcement by these two biomaterials should be considered safe from the calcification point of view.     

Ethanol selectively potentiates GABA-mediated inhibition of single feline cortical neurons

Ethanol (alcohol) released from micropipettes by electro-osmosis (up to 10 nA from 0.3 M in 165 mM NaCl solution) potentiated the inhibition of firing of single cortical neurons produced by iontophoretically-applied pulses of γ-aminobutyric acid (GABA), whereas it had no effect or a mild antagonistic effect on the inhibition produced by pulses of glycine, and had an antagonistic effect on the inhibition produced by pulses of serotonin or dopamine. The potentiation of iontophoretically-applied GABA was also obtained by intravenously-applied ethanol (0.2–2 mg/kg). Furthermore, ethanol applied by electro-osmosis or intravenously in the same doses potentiated the inhibition of firing of single cortical neurons evoked by electrical stimulation of the surface of the cerebral cortex, which is believed to be mediated by endogenous GABA. These findings may have implications for alcoholism, since GABAergic neurotransmission is involved in the mechanism of action of anxiolytic drugs and anxiety is involved in the etiology of alcoholism.     

Preparation and Toxicological Assessment of Functionalized Carbon Nanotube-Polymer Hybrids

Novel Carbon Nanotube-Polymer Hybrids were synthesized as potential materials for the development of membranes for water treatment applications in the field of Membrane Bioreactors (MBRs). Due to the toxicological concerns regarding the use of nanomaterials in water treatment as well as the rising demand for safe drinking water to protect public health, we studied the functionalization of MWCNTs and Thin-MWCNTs as to control their properties and increase their ability of embedment into porous anisotropic polymeric membranes. Following the growth of the hydrophilic monomer on the surface of the properly functionalized CNTs, that act as initiator for the controlled radical polymerization (ATRP) of sodium styrene sulfonate (SSNa), the antimicrobial quaternized phosphonium and ammonium salts were attached on CNTs-g-PSSNa through non-covalent bonding. In another approach the covalent attachment of quaternized ammonium polymeric moieties of acrylic acid-vinyl benzyl chloride copolymers with N,N-dimethylhexadecylamine (P(AA12-co-VBCHAM)) on functionalized CNTs has also been attempted. Finally, the toxicological assessment in terms of cell viability and cell morphological changes revealed that surface characteristics play a major role in the biological response of functionalized CNTs.     

Characterization of Inducible Models of Tay-Sachs and Related Disease

Tay-Sachs and Sandhoff diseases are lethal inborn errors of acid β-N-acetylhexosaminidase activity, characterized by lysosomal storage of GM2 ganglioside and related glycoconjugates in the nervous system. The molecular events that lead to irreversible neuronal injury accompanied by gliosis are unknown; but gene transfer, when undertaken before neurological signs are manifest, effectively rescues the acute neurodegenerative illness in Hexb−/− (Sandhoff) mice that lack β-hexosaminidases A and B. To define determinants of therapeutic efficacy and establish a dynamic experimental platform to systematically investigate cellular pathogenesis of GM2 gangliosidosis, we generated two inducible experimental models. Reversible transgenic expression of β-hexosaminidase directed by two promoters, mouse Hexb and human Synapsin 1 promoters, permitted progression of GM2 gangliosidosis in Sandhoff mice to be modified at pre-defined ages. A single auto-regulatory tetracycline-sensitive expression cassette controlled expression of transgenic Hexb in the brain of Hexb−/− mice and provided long-term rescue from the acute neuronopathic disorder, as well as the accompanying pathological storage of glycoconjugates and gliosis in most parts of the brain. Ultimately, late-onset brainstem and ventral spinal cord pathology occurred and was associated with increased tone in the limbs. Silencing transgenic Hexb expression in five-week-old mice induced stereotypic signs and progression of Sandhoff disease, including tremor, bradykinesia, and hind-limb paralysis. As in germline Hexb−/− mice, these neurodegenerative manifestations advanced rapidly, indicating that the pathogenesis and progression of GM2 gangliosidosis is not influenced by developmental events in the maturing nervous system.