The influence of oxygen on the survival and yield of DNA double-strand breaks in irradiated yeast cells.

Survival and induction of DNA double-strand breaks were studied in cells of Saccharomyces cerevisiae irradiated under oxic or anoxic conditions with 30 MeV electrons. A linear relationship between DNA double-strand breakage and dose was found in both cases. The o.e.r.-value for colony forming ability was found to be 1.9 +/- 0.2, whereas the o.e.r.-value for DNA double-strand breakage was 3.0 +/- 0.1. These results are not inconsistent with the idea that DNA double-strand breaks are involved in killing of yeast cells. The frequency of induction of DNA double-strand breaks was found to be 0.74 x 10(-11) double-strand breaks per g/mol per Gy when cells were irradiated under oxygen and 0.24 x 10(-11) double-strand breaks per g/mol per Gy under nitrogen.     

Applications of self-organizing map for patterning macrofungal diversity of xerothermic swards

Macrofungal communities were investigated in four associations of xerothermic swards: Festucetum pallentis, Origano-Brachypodietum, Adonido-Brachypodietum pinnati and Diantho-Armerietum elongatae in a Jurassic area of the Częstochowa Upland (southern Poland). A total of 47 species were recorded. The self-organising map (SOM)—an unsupervised algorithm for artificial neural networks—was used to recognise patterns in the macrofungal communities of diverse xerothermic swards. Only two associations were mycologically similar: Origano-Brachypodietum and Adonido-Brachypodietum pinnati. Species with high and significant IndVal (the species indicator value) for each investigated phytocoenoses are presented. The presence of macrofungal species and the participation of indicator species were connected with habitat factors of plant associations, as documented by the IndVal application. In the least fertile phytocoenoses, macrofungal communities were poor with few indicator species. The more fertile phytocoenoses had richer and more varied communities of macrofungi with higher numbers of indicator species. The ordering methods applied in this study were very effective for analyzing the macrofungal communities existing in plant associations.     

Toxicity of extracellular secreted alpha-synuclein: Its role in nitrosative stress and neurodegeneration

It has been demonstrated that both oligomerisation and accumulation of α-synuclein (ASN) are the key molecular processes involved in the pathophysiology of neurodegenerative diseases such as Parkinson’s disease, Alzheimer’s disease and other synucleinopathies. Alterations of ASN expression and impairment of its degradation can lead to the formation of intracellular deposits of this protein, called Lewy bodies. Overexpressed or misfolded ASN could be secreted to the extracellular space. Today the prion-like transmission of ASN oligomers to neighbouring cells is believed to be responsible for protein modification and propagation of neurodegeneration in the brain. It was presented that oxidative/nitrosative stress may play a key role in ASN secretion and spread of ASN pathology. Moreover, ASN-evoked protein oxidation, nitration and nitrosylation lead to disturbances in synaptic transmission and cell death. The interaction of secreted ASN with other amyloidogenic proteins and its involvement in irreversible mitochondrial disturbances and oxidative stress were also described. A better understanding of the mechanisms of ASN secretion and dysfunction may help to explain the molecular mechanisms of neurodegeneration and may be the basis for the development of novel therapeutic strategies.     

Transglutaminase 6: a protein associated with central nervous system development and motor function

Transglutaminases (TG) form a family of enzymes that catalyse various post-translational modifications of glutamine residues in proteins and peptides including intra- and intermolecular isopeptide bond formation, esterification and deamidation. We have characterized a novel member of the mammalian TG family, TG6, which is expressed in a human carcinoma cell line with neuronal characteristics and in mouse brain. Besides full-length protein, alternative splicing results in a short variant lacking the second β-barrel domain in man and a variant with truncated β-sandwich domain in mouse. Biochemical data show that TG6 is allosterically regulated by Ca²⁺ and guanine nucleotides. Molecular modelling indicates that TG6 could have Ca²⁺ and GDP-binding sites related to those of TG3 and TG2, respectively. Localization of mRNA and protein in the mouse identified abundant expression of TG6 in the central nervous system. Analysis of its temporal and spatial pattern of induction in mouse development indicates an association with neurogenesis. Neuronal expression of TG6 was confirmed by double-labelling of mouse forebrain cells with cell type-specific markers. Induction of differentiation in mouse Neuro 2a cells with NGF or dibutyryl cAMP is associated with an upregulation of TG6 expression. Familial ataxia has recently been linked to mutations in the TGM6 gene. Autoantibodies to TG6 were identified in immune-mediated ataxia in patients with gluten sensitivity. These findings suggest a critical role for TG6 in cortical and cerebellar neurons.     

Differences in editing of mitochondrial nad3 transcripts from CMS and fertile carrots

A high level of the nucleotide sequence conservation was found for mitochondrial nad3 gene of carrot. Three silent nucleotide substitutions differentiate nad3 open reading frames from cytoplasmic male sterile and male fertile carrots. All these differences are preserved on the RNA level. Partial and silent editing also distinguished both carrots. Three of the C to U conversions were specific to the fertile line. In the two examined carrot lines editing did not affect the mode of alteration of encoded amino acids.     

Surface plasmon resonance (SPR) as a tool for antibody conjugate analysis

Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin (4) and anti-fluorescein (5) monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1. Only minor changes in the apparent equilibrium dissociation constants of the antibody conjugates for their ligands resulted from the conjugation process. However, comparison of the initial binding rate of the conjugates with their ligands with those of the unmodified antibodies over a range of concentrations showed that the antibody conjugates were partially inactivated. The anti-fluorescein conjugates retained at least 90% of their immunoreactivity over the range of modification tested, while anti-biotin conjugates showed a progressive loss of reactivity with increased substitution by the label.     

Intrinsic factor-mediated modulation of cyanocobalamin-N-sulfonyl-acridinium-9-carboxamide chemiluminescence

Two cyanocobalamin-N-sulfonyl-acridinium-9-carboxamides with linkage through the N(10) or 9-position were prepared from B(12)-e-carboxylic acid. The noncovalent association of intrinsic factor with these ligands resulted in specific modulation of the associated chemiluminescence signal either by quenching or changing the emission profile. Either effect was sufficient to formulate a homogeneous assay to detect vitamin B(12) in buffer.