Copy number variations of genes involved in stress responses reflect the redox state and DNA damage in brewing yeasts

The yeast strains of the Saccharomyces sensu stricto complex involved in beer production are a heterogeneous group whose genetic and genomic features are not adequately determined. Thus, the aim of the present study was to provide a genetic characterization of selected group of commercially available brewing yeasts both ale top-fermenting and lager bottom-fermenting strains. Molecular karyotyping revealed that the diversity of chromosome patterns and four strains with the most accented genetic variabilities were selected and subjected to genome-wide array-based comparative genomic hybridization (array-CGH) analysis. The differences in the gene copy number were found in five functional gene categories: (1) maltose metabolism and transport, (2) response to toxin, (3) siderophore transport, (4) cellular aldehyde metabolic process, and (5) L-iditol 2-dehydrogenase activity (p < 0.05). In the Saflager W-34/70 strain (Fermentis) with the most affected array-CGH profile, loss of aryl-alcohol dehydrogenase (AAD) gene dosage correlated with an imbalanced redox state, oxidative DNA damage and breaks, lower levels of nucleolar proteins Nop1 and Fob1, and diminished tolerance to fermentation-associated stress stimuli compared to other strains. We suggest that compromised stress response may not only promote oxidant-based changes in the nucleolus state that may affect fermentation performance but also provide novel directions for future strain improvement.     

Spatial and temporal distribution of copepod communities in the Adriatic Sea

The distribution of planktonic copepods in the Adriatic Seahas been analyzed on the basis of 132 samples collected at 35fixed stations during 4 seasonal cruises. A total of 127 specieshave been determined and 3 characteristic copepod communitieshave been identified. The distribution of copepod species hasbeen found to reflect the dual physiognomy of the Adriatic.The shallow northern section is characterized by high densityvalues, low species diversity, and the dominance of speciesbelonging to the estuarine and coastal communities. The relativelydeep waters of the southern section are characterized by lowdensity values, high species diversity, and the presence ofmany species belonging to the upper, middle, and lower zonesof the oceanic community. Temporal variations in the horizontalspread of these 3 communities are discussed in relation to seasonalcycles in abundance, vertical migration patterns, and the currentregime in the Adriatic.     

Polymerase eta is a short-lived, proteasomally degraded protein that is temporarily stabilized following UV irradiation in Saccharomyces cerevisiae

Saccharomyces cerevisiae Rad30 is the homolog of human DNA polymerase eta whose inactivation leads to the cancer-prone syndrome xeroderma pigmentosum variant. Both human and yeast polymerase eta are responsible for error-free bypass of UV-induced cis-syn pyrimidine dimers and several other DNA lesions. Here we show, using yeast strains expressing TAP-tagged Rad30, that the level of this protein is post-translationally regulated via ubiquitination and proteasome-mediated degradation. The half-life of Rad30 is 20 min and it increases due to proteasomal defects. Mutations inactivating components of the Skp1/cullin/ F-box (SCF) ubiquitin ligase complex: Skp1 and the F-box protein Ufo1 stabilize Rad30. Our results indicate also that ultraviolet irradiation causes transient stabilization of Rad30, which leads, in turn, to temporary accumulation of this polymerase in the cell. We conclude that proteolysis plays an important role in regulating the cellular abundance of Rad30. These results are the first indication of a role for controlled proteasomal degradation in modulating cellular level of translesion DNA polymerase in eukaryotes.     

In vitro morphogenetic responses from obligatory apomictic Taraxacum belorussicum Val. N. Tikhom seedlings explants

Taraxacum belorussicum Val. N. Tikhom, a poorly known and obligatory apomictic species, is an attractive plant material for studying the embryological, genetic and molecular mechanisms of apomixis. This work aims to obtain an efficient protocol for Taraxacum belorussicum regeneration. Four types of explants (cotyledons, hypocotyls, meristems and roots) that were taken from 2-weeks-old seedlings were used for in vitro cultures, and a fast and efficient protocol of T. belorussicum regeneration was obtained. Various ½ MS-based media containing IAA (5.71 µM), TDZ (4.54 µM) and PSK (100 nM) were chosen to assess the morphogenetic abilities of selected T. belorussicum explants. Studies on the role of PSK were done in three independent experiments, where the most significant factors were always light and darkness. All explants produced callus by the third day of culture and adventitious shoots after 7 days, although in an asynchronous indirect manner, and with different intensities for all explant types. The most preferred medium culture for hypocotyl, cotyledon and meristem explants was ½ MS?+?TDZ, and ½ MS?+?IAA?+?TDZ?+?PSK for roots which were the only explant sensitive to PSK. A short darkness pretreatment (8 days) in PSK medium was found suitable to enhance organogenesis. Secondary organogenesis was observed for regenerated plants on meristem explants from the ½ MS?+?IAA?+?TDZ?+?PSK medium. A weak somatic embryogenesis was observed for hypocotyl and cotyledon explants from ½ MS?+?IAA?+?TDZ and ½ MS?+?IAA?+?TDZ?+?PSK media. Histological and scanning electron microscope images (SEM) of T. belorussicum confirmed indirect organogenesis and somatic embryogenesis. Plant material treated with aniline blue solution revealed the presence of callose in the cell walls of cotyledon and hypocotyl explants. The presence of extracellular matrix (ECM) and heterogenic structure of callus was also verified by scanning electron microscopy and light microscopy, confirming the high morphogenetic ability of T. belorussicum.

     

Prevalence of oral hairy leukoplakia and epithelial infection by Epstein-Barr virus in pregnant women and diabetes mellitus patients--cytopathologic and molecular study

Oral hairy leukoplakia (OHL) is generally reported in patients with severe immunosuppression, except for a few cases in individuals with moderate degree of immunodeficiency. It is a white lesion that appears mainly in the lateral border of the tongue, caused by Epstein-Barr virus (EBV). The nuclear changes caused by EBV (Cowdry A inclusion, ground glass and nuclear beading), observed in cytopathology, are specific and enough for the definitive diagnosis of OHL, independent of the identification of the virus. Here we investigated the prevalence of OHL and the presence of EBV-DNA in the lateral borders of the tongue from 90 pregnant women, 90 diabetes mellitus (DM) patients, 30 healthy individuals (negative group) and 30 HIV+ with OHL (positive group). Smears were analyzed by cytopathology and polymerase chain reaction (PCR). A case of subclinical OHL and candidiasis was identificated in a DM patient by cytopathologic analysis. PCR results demonstrated EBV-DNA in 65% of the pregnant women, in 35% of DM patients, and in 20% of the healthy individuals. We concluded that DM patients can develop OHL with a low prevalence. Furthermore, the prevalence of the EBV in lateral border of the tongue is larger in pregnant women than in healthy individuals.     

Chronic intermittent but not constant hypoxia decreases NAA/Cr ratios in neonatal mouse hippocampus and thalamus

Chronic constant hypoxia (CCH) and chronic intermittent hypoxia (CIH) are known to have deleterious effects on the central nervous system. Because of the difference in the pattern of hypoxic exposure, it is possible that the pathological outcome would vary. The N-acetyl aspartate/creatine (NAA/Cr) ratio is a reliable marker of neuronal integrity, and this can be noninvasively measured by proton nuclear magnetic resonance spectroscopy. P2 CD1 mouse pups with their dams were exposed to either CCH, where the Fi(O(2)) was maintained at 11% continuously or to CIH, where the Fi(O(2)) was varied between 21 and 11% every 4 min. P30 mice exposed to intermittent hypoxia for 4 wk demonstrated a significant decrease in the NAA/Cr ratio in the hippocampus and thalamus, which was reversed by a subsequent exposure to 4 wk of normoxia. Meanwhile, mice exposed to 4 wk of constant hypoxia did not demonstrate any differences in their NAA/Cr ratios from controls in these brain regions. These results indicate that an intermittent pattern of hypoxic exposure may have a more adverse effect on neuronal function and integrity than a continuous one. The reversal of NAA/Cr levels to baseline during the return to normoxia indicates that therapeutic strategies targeted at alleviating the intermittent hypoxic stress in diseases, such as obstructive sleep apnea, have the potential for inducing significant neurocognitive recovery in these patients.     

Saccharomyces cerevisiae Hsp31p, a stress response protein conferring protection against reactive oxygen species

The Saccharomyces cerevisiae HSP31 (YDR533c) gene encodes a protein that belongs to the DJ-1/PfpI family and its function is unknown. Homologs to Hsp31p polypeptide can be found in organisms from all systematic groups of eukaryotes and prokaryotes, and the functions of the vast majority of them are also hypothetical. One of the homologs is human protein DJ-1. Various amino acid substitutions within this protein correlate with early onset hereditary Parkinson's disease. The deletion of the HSP31 gene displays no apparent phenotype under standard growth conditions, but a thorough functional analysis of S. cerevisiae revealed that its absence makes the cells sensitive to a subset of reactive oxygen species (ROS) generators. HSP31 is induced under conditions of oxidative stress in a YAP1-dependent manner. Similar to other stress response genes, it is also induced in the postdiauxic phase of growth and this induction is YAP1-independent. The patterns of sensitivities to various ROS generators of the hsp31Delta strain and the strain with the deletion of SOD1, another gene defending the cell against ROS, are different. We postulate that Hsp31p protects the cell against oxidative stress and complements other stress protection systems within the cell.     

Revisiting the phylogeography of Asellus aquaticus in Europe: insights into cryptic diversity and spatiotemporal diversification

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Preventing horticultural introductions of invasive plants: potential efficacy of voluntary initiatives

Although prevention is the most cost-effective way to avoid the enormous expenses associated with plant invasions, invasive plants continue to be imported as trade commodities for horticultural use. With very little government regulation of horticultural imports of invasive plants, efforts have turned toward fostering voluntary initiatives to encourage self-regulation by the horticulture trade. Our study takes the first step toward evaluating the potential success of these voluntary initiatives. We conducted a survey of nursery professionals to gauge their perceptions of invasive species, the role of the horticulture trade in invasive plant introductions, and their participation—potential and actual—in preventive measures outlined in the St Louis Voluntary Codes of Conduct for nursery professionals. We found nursery professionals to be highly aware of invasive plants and to accept responsibility as a trade for horticultural introductions. Although only 7% of respondents had heard of the St Louis Voluntary Codes of Conduct, the majority (57%) reported having participated in at least two of seven preventive measures, and most (78%) reported willingness to engage in the majority of preventive measures. We found that several factors significantly predict increased participation in preventive measures, particularly awareness of invasive plants and involvement in trade associations. We also identified incentives and obstacles to participating in preventive behaviors, including “concern for the environment” and “lack of information,” respectively. Our results suggest that participation in voluntary initiatives will improve through increased outreach, and we provide specific recommendations for improving participation in voluntary programs in the horticulture trade.     

The Influence of Invertebrate Predators on Daphnia Spatial Distribution and Survival in Laboratory Experiments: Support for Daphnia Horizontal Migration in Shallow Lakes

The behavioural response of Daphnia cucullata to the presence of the pelagic invertebrate predator Leptodora kindtii, and the predation rate of littoral dragonfly nymphs on this species were investigated under laboratory conditions. Results of this study revealed a strong hiding response of Daphnia cucullata in the presence of the predatory cladoceran, L. kindtii, which was similar to the response of Daphnia in the presence of juvenile perch. This suggests that pelagic invertebrate predators may cause Daphnia to hide in the littoral zone which could result in increased exposure to predation by littoral invertebrates. A strong influence of dragonfly nymphs on D. cucullata, both in the presence and absence of macrophytes, was found. The average predation rate of Odonata larvae was about 5 prey ind^–1 h^–1 and did not differ significantly between treatments. Quantification of dragonfly pressure on Daphnia populations will require cross‐verification with field experiments since in the natural conditions Daphnia seeks a shelter in the vegetation stands against predation by Leptodora, despite the occurrence of odonates. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)