New Saccharomyces cerevisiae baker's yeast displaying enhanced resistance to freezing

Three procedures were used to obtain new Saccharomyces cerevisiae baker's yeasts with increased storage stability at -20, 4, 22, and 30 degrees C. The first used mitochondria from highly ethanol-tolerant wine yeast, which were transferred to baker's strains. Viability of the heteroplasmons was improved shortly after freezing. However, after prolonged storage, viability dramatically decreased and was accompanied by an increase in the frequency of respiratory-deficient (petite) mutant formation. This indicated that mitochondria were not stable and were incompatible with the nucleus. The strains tested regained their original resistance to freezing after recovering their own mitochondria. The second procedure used hybrid formation after protoplast fusion and isolation on selective media of fusants from baker's yeast meiotic products resistant to parafluorphenylalanine and cycloheximide, respectively. No hybrids were obtained when using the parentals, probably due to the high ploidy of the baker's strains. Hybrids obtained from nonisogenic strains manifested in all cases a resistance to freezing intermediate between those of their parental strains. Hybrids from crosses between meiotic products of the same strain were always more sensitive than their parentals. The third method was used to develop baker's yeast mutants resistant to 2-deoxy-d-glucose (DOG) and deregulated for maltose and sucrose metabolism. Mutant DOG21 displayed a slight increase in trehalose content and viability both in frozen doughs and during storage at 4 and 22 degrees C. This mutant also displayed a capacity to ferment, under laboratory conditions, both lean and sweet fresh and frozen doughs. For industrial uses, fermented lean and sweet bakery products, both from fresh and frozen doughs obtained with mutant DOG21, were of better quality with regard to volume, texture, and organoleptic properties than those produced by the wild type.     

Inhibitory effect of a glycoprotein isolated from golden oyster mushroom ( Pleurotus citrinopileatus ) on the lipopolysaccharide-induced inflammatory reaction in RAW 264.7 macrophage

Mushrooms have become an important source of natural antitumor, antiviral, antibacterial, immunomodulatory, and anti-inflammatory agents. Golden oyster mushroom, Pleurotus citrinopileatus , is a common mushroom in oriental countries for human consumption. The present study investigated the anti-inflammatory reaction of the bioactive nonlectin glycoprotein (PCP-3A) isolated from the fresh fruiting body of this mushroom. Western blot analysis on LPS-induced iNOS, COX-2, and NF-κB expressions in RAW 264.7 cells as affected by PCP3-A was performed to elucidate the mechanism of NO and PGE2 reduction. The results showed that PCP-3A failed to affect RAW 264.7 viability at a concentration up to 6.25 μg/mL, but inhibited LPS (1 μg/mL)-induced expression, and that PCP-3A inhibited the production of NO and PGE2 in LPS-activated macrophages via the down-regulation of certain pro-inflammatory mediators, including iNOS and NF-κB.     

Conservation planning of vertebrate diversity in a Mediterranean agricultural-dominant landscape

To improve effectiveness of protected areas, selection of priority areas should include consideration of three main components, namely special conservation elements, focal species and representation. We present a three-track approach related to these components for vertebrate conservation planning in Castilla-La Mancha, central Spain. As special conservation elements, we identified Priority Areas for Conservation of species using five criteria: species richness, geographic rarity, species vulnerability, a Combined Index of these three criteria, and a Standardised Biodiversity Index (SBI) that integrate the three criteria and four studied taxa. The Natura 2000 Network was used to include conservation areas for focal species. We evaluated the representation of every landscape type in the existing conservation areas. To delineate the spatial configuration for vertebrate conservation, we combined the identified Priority Areas for Conservation, existing conservation areas and connectivity areas by cost–distance analysis. The Combined Index was the most efficient criterion analyzed to identify Priority Areas for Conservation. The Natura 2000 Network showed a high percentage of coincidence with identified Priority Areas for Conservation, whereas the natural protected areas network had a low percentage of coincidence. Six agricultural landscapes were inadequately represented in the current conservation network. According to our multi-track approach, ∼29% of study area was required to capture 100% of vertebrate species and all landscape types. Our results show that the existing conservation areas are insufficient to guarantee the conservation of biodiversity in the study region. Additional areas with outstanding features of diversity, connectivity areas, and establishment of targets for off-reserve conservation are of fundamental importance for strengthening biodiversity conservation.     

Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis

Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.     

Expression of TGFBR1, TGFBR2, TGFBR3, ACVR1B and ACVR2B is altered in ovaries of cows with cystic ovarian disease

The objective of this study was to examine the expression of transforming growth factor beta receptor (TGFBR)1, TGFBR2, TGFBR3, activin receptor (ACVR)1B and ACVR2B in ovaries of cows with cystic ovarian disease (COD). The expression of the selected receptors was determined by immunohistochemistry in sections of ovaries from cows with ACTH‐induced and spontaneous COD. Expression of TGFBR1 and TGFBR3 was higher in granulosa cells of cysts from cows with spontaneous COD than in tertiary follicles from the control group. Additionally, TGFBR3 expression was higher in granulosa cells of cysts from cows with ACTH‐induced COD than in those from the control group and lower in theca cells of spontaneous and ACTH‐induced cysts than in tertiary control follicles. There were no changes in the expression of TGFBR2. ACVR1B expression was higher in granulosa cells of tertiary follicles of cows with spontaneous COD than in the control group, whereas ACVR2B expression was higher in cysts of the spontaneous COD group than in tertiary follicles from the control group. The alterations here detected, together with the altered expression of the ligands previously reported, indicate alterations in the response of the ligands in the target cells, modifying their actions at cellular level.     

Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: modifications induced by oxidation and suitable markers of oxidative status

Modifications of virgin olive oil subjected to accelerated storage were evaluated by HS-SPME analysis. To find a suitable marker of oxidative degradation, the volatile compounds showing variable concentration during the oxidative process have been identified and quantified by SPME coupled to GC-MS and GC-FID, respectively. The SPME analysis results were then compared with the parameters usually applied to assess the oxidative status of lipids, such as peroxide value, spectrophotometric absorbance, and loss of unsaturated fatty acids. Finally, the assessment of nonanal has been suggested as a marker of oxidative degradation. This rapid, inexpensive, and reliable method may allow screening of oils prior to testing by a panel of assessors.     

Soil reinforcement by the roots of six dominant species on eroded mountainous marly slopes (Southern Alps,France)

In marly catchments of the French Southern Alps, the development of plant root systems is essential to increase slope stability and mitigate soil erosion, prevalent in this area. In a context of land restoration, it is important to be able to evaluate plant efficiency for soil reinforcement. This paper presents the results of investigations carried out on six dominant species from marly gullies. It aims to compare the additional soil cohesion they provide at the early stages of their development. The six following species were collected: two tree species, Pinus nigra and Quercus pubescens, two shrubby species, Genista cinerea and Thymus serpyllum, and two herbaceous species, Achnatherum calamagrostis and Aphyllantes monspeliensis. For each of them, we measured root tensile strength and root area ratio in order to calculate the potential root reinforcement and to compare species suitability to prevent shallow mass movements. Results showed significant differences between species. The herbaceous species A. monspeliensis and the shrubby species G. cinerea provided the highest increase in soil shear strength while the tree species, P. nigra and Q. pubescens were the least efficient. These results, along with the knowledge on vegetation dynamics and species response to erosive constraint, allow us to better evaluate land vulnerability to erosion and the efficiency of restoration actions in eroded marly lands.