Pools of water in anhydrobiotic organisms: A thermally stimulated depolarization current study

The ability to survive the removal of water in anhydrous biosystems is especially remarkable as a departure from the manifold structural and functional dependences on the presence of H₂O molecules. Identifiable pools of water present in dry soybean axes were investigated by means of the thermally stimulated depolarization current method. Samples were examined in the temperature range 100-340 K and over water contents (h, in gram H₂O per gram sample dry weight) ranging from h = 0.05 to 0.30 g/g. Three water-dependent relaxation mechanisms were detected; one attributed to dipolar reorientation of H₂O molecules hydrogen-bonded to other water molecules, one to reorientation of CH₂OH groups, and one to a glass transition in sugar-water domains. These glassy domains can protect intracellular components against destruction in the dehydrated state. Interestingly, protecting glassy domains were not found in dehydration intolerant seeds, supporting the hypothesis that the ability to withstand dehydration is associated with intracellular glass formation. A model for the state of cell water at interfaces is proposed.     

Glass transitions in soybean seed : relevance to anhydrous biology

We have investigated the mechanism by which anhydrobiotic organisms can survive severe dehydration. The method used was measurement of the rotational diffusion coefficient of a hydrophilic spin probe, inserted in the cytoplasm of soybean (Glycine max L.) axes, as a function of temperature and sample water content. Results indicate the existence of a hydration-dependent glass-like transition at physiological temperatures. No glass transitions have been observed in desiccation-intolerant samples, suggesting that the ability to withstand dehydration is associated with glass formation.     

d(GATC) sequences influence Escherichia coli mismatch repair in a distance-dependent manner from positions both upstream and downstream of the mismatch

The role of d(GATC) sites in determining the efficiency of methyl-directed mismatch repair in Escherichia coli was investigated. Transfection of host bacteria, both proficient and deficient in mismatch repair, with a series of artificially constructed M13 heteroduplexes showed that a decrease in the total number of d(GATC) sequences within these vectors lowered the efficiency of repair in vivo. Single hemimethylated d(GATC) sequences were still able to direct the correction event to the unmethylated strand, providing that the mismatch to d(GATC) site distance was shorter than approximately 1 kb. In excess of this distance, the effect of hemimethylated d(GATC) sites on mismatch correction was almost unnoticeable. The directionality of the repair event could be dictated by d(GATC) sequences situated both upstream and downstream of the mispair, suggesting that this important antimutagenic pathway can proceed bidirectionally.     

Synthesis of diacylglycerol de novo is responsible for permanent activation and down-regulation of protein kinase C in transformed cells

We measured the synthesis of diacylglycerol de novo in normal NIH/3T3 fibroblasts and in cells transformed by ras, src, sis and abl oncogenes. Analysis of the incorporation of glucose-derived 14C into diacylglycerol indicated that neosynthesis of diacylglycerol was constitutively active in the transformed cell lines. Elevated levels of diacylglycerol and persistent activation/down-regulation of protein kinase C reduced the binding of phorbol dibutyrate to transformed cells. This phenomenon could be reversed by blocking the glycolytic pathway, thus indicating that neosynthesized diacylglycerol was responsible for persistent activation and down-regulation of protein kinase C. In transformed cells, protein kinase C activity could not be stimulated by the addition of diolein; however, inhibition of glycolysis restored the ability of transformed cells to respond to diolein. Taken together these data indicate that constitutive synthesis of diacylglycerol de novo is responsible for activation and down-regulation of protein kinase C in transformed cells, and it may play a role in altered mitogenic signalling.     

In vitro efficacy of the novel peptide CECT7121 against bacteria isolated from mastitic dairy cattle

Aims:  To evaluate the in vitro bactericidal activity of the novel antimicrobial peptide (AP) CECT7121 against Gram-positive bacteria from mastitic dairy cattle.
Methods and Results:  A total of 15 Staphylococcus aureus , 10 Streptococcus dysgalactiae , 7 Strep. uberis , 1 Strep. agalactiae strains were isolated from 33 different mastitic dairy cattle, sourced from two dairies in Tandil-Argentina. Isolates from each of the bacterial species screened which developed the lowest inhibition zones in response to the peptide, were further evaluated in a series of time-killing curve studies. No survivors were detected in whole strains (from the three Streptococcal species isolated) within 120 min of incubation in presence of the peptide. The Staph. aureus isolates were less sensitive but, nevertheless, a drop in viable counts to below the detection limit was achieved for all the test strains by the final postincubation sampling point at 180 min.
Conclusions:  The study demonstrated the in vitro efficacy of the AP-CECT7121 against a variety strains of Gram positives isolated from mastitic dairy cattle.
Significance and Impact of the Study:  There is urgent global interest in the development of natural alternatives for the control and prevention of mastitis. Confirmation of the in vitro activity of the novel AP-CECT7121 against Gram-positive isolates encourages further research.     

Aerobiological monitoring ofAlternaria fungal spores: a comparison between surveys in 1992 and 1993 and local meteorological conditions

A study of airborneAlternaria spores was carried out over the period from April 1992 to October 1993, in the Rehabilitation Medical Centre of Montescano (PV), using an automatic volumetric spore trap (VPPS 2000 Lanzoni). The evaluation of the air concentration of these spores is becoming ever more important as it is this mycophyte which is the commonest cause of a positive allergy test. In 1993 the concentration ofAlternaria spores was much higher than in the previous year and the period of highest sporulation was different, depending on the different rainfall pattern of the 2 years.     

Correlation between cardiac oxidative stress and myocardial pathology due to acute and chronic norepinephrine administration in rats

BACKGROUND: To investigate the cardiotoxic role of reactive oxygen species (ROS) and of products derived from catecholamines auto-oxidation, we studied: (1) the response of antioxidant cardiac cellular defence systems to oxidative stress induced by norepinephrine (NE) administration, (2) the effect of NE administration on cardiac beta1-adrenergic receptors by means of receptor binding assay, (3) the cellular morphological alterations related to the biologically cross-talk between the NE administration and cytokines [tumor necrosis factor-alpha (TNF-alpha), monocyte chemotactic protein-1 (MCP-1), interleukins IL6, IL8, IL10]. METHODS AND RESULTS: A total of 195 male rats was used in the experiment. All animals underwent electrocardiogram (EKG) before being sacrificed. The results obtained show that NE administration influences the antioxidant cellular defence system significantly increasing glutathione peroxidase (GPx) activity, glutathione reductase (GR) and superoxide dismutase (SOD). The oxidized glutathione (GSH/GSSG) ratio significantly decreases and malondialdehyde (MDA) levels increase showing a state of lipoperoxidation of cardiac tissue. We describe a significant apoptotic process randomly sparse in the damaged myocardium and the effect of ROS on the NE-mediated TNF-alpha, MCP-1, and IL6, IL8, IL10 production. CONCLUSIONS: Our results support the hypothesis that catecholamines may induce oxidative damage through reactive intermediates resulting from their auto-oxidation, irrespective of their interaction with adrenergic receptors, thus representing an important factor in the pathogenesis of catecholamines-induced cardiotoxicity. The rise of the cardioinhibitory cytokines may be interpreted as the adaptive response of jeopardized myocardium with respect to the cardiac dysfunction resulting from NE injection.