Techniques for the detection of pathogenic Cryptococcus species in wood decay substrata and the evaluation of viability in stored samples

In this study, we evaluated several techniques for the detection of the yeast form of Cryptococcus in decaying wood and measured the viability of these fungi in environmental samples stored in the laboratory. Samples were collected from a tree known to be positive for Cryptococcus and were each inoculated on 10 Niger seed agar (NSA) plates. The conventional technique (CT) yielded a greater number of positive samples and indicated a higher fungal density [in colony forming units per gram of wood (CFU.g^-1) ] compared to the humid swab technique (ST). However, the difference in positive and false negative results between the CT-ST was not significant. The threshold of detection for the CT was 0.05.10³ CFU.g^-1, while the threshold for the ST was greater than 0.1.10³ CFU^-1. No colonies were recovered using the dry swab technique. We also determined the viability of Cryptococcus in wood samples stored for 45 days at 25ºC using the CT and ST and found that samples not only continued to yield a positive response, but also exhibited an increase in CFU.g^-1, suggesting that Cryptococcus is able to grow in stored environmental samples. The ST.1, in which samples collected with swabs were immediately plated on NSA medium, was more efficient and less laborious than either the CT or ST and required approximately 10 min to perform; however, additional studies are needed to validate this technique.     

Anti-phospholipase C antibodies inhibit the lectin-induced proliferation of human lymphocytes

A novel approach was used to assess the role of phosphoinositide hydrolysis in the mitogenic action of phytohemagglutinin (PHA) or concanavalin A (ConA). The treatment of human peripheral blood leukocytes (PBL) with monospecific antibodies against phospholipase C (PLC) produced a dose-dependent inhibition (up to 100%) of PHA (10 g/ml) or ConA (25 g/ml) proliferative effects. Thus, the activation of membrane-bound PLC is asine-qua-non condition for lectin-induced proliferation of T lymphocytes. The key-role of PLC versus protein kinase C (PKC) is stressed by the fact that the inhibition of PKC with Hidaka's compound H-7 (40 M) produced only a partial blockade (about 25%) of lectin mitogenic effect.To whom correspondence should be addressed.     

The immediate-early growth response in regenerating liver and insulin-stimulated H-35 cells: comparison with serum-stimulated 3T3 cells and identification of 41 novel immediate-early genes.

Liver regeneration provides a unique system for analysis of mitogenesis in intact, fully developed animals. Cellular immediate-early genes likely play an important role in cell cycle regulation and have been extensively studied in mitogen-stimulated fibroblasts lymphocytes but not in liver. We have begun to characterize the immediate-early growth response genes of mitogen-stimulated liver cells, specifically, regenerating liver and insulin-stimulated Reuber H-35 hepatoma cells, and to address differences in growth response between different cell types. Through subtraction and differential screening of cDNA libraries from regenerating liver and insulin-treated H-35 cells, we have extensively characterized 341 differentially expressed clones and identified 52 immediate-early genes. These genes have been partially sequenced and subjected to Northern (RNA) blot analysis, and 41 appear to be novel. Surprisingly, two-thirds of these genes are also expressed in BALB/c 3T3 cells, but only 10 were identified in previous studies of 3T3 cells, and of these, 6 include well-known genes like jun and fos, and only 4 are novel. Approximately one-third of the immediate-early genes identified in mitogen-stimulated liver cells or serum-stimulated NIH 3T3 cells are expressed in a tissue-specific fashion, indicating that cell type-specific regulation of the proliferative response occurs during the immediate-early period. Our findings indicate that the immediate-early response is unusually complex for the first step in a regulatory cascade, suggesting that multiple pathways must be activated. The abundance of immediate-early genes and the highly varied pattern of their expression in different cell types suggest that the tissue specificity of the proliferative response arises from the particular set of these genes expressed in a given tissue.     

Classification of types of ontogenetic reproduction

The object of this paper is to present an original classification of ontogenetic reproduction. The main general criterion used is the degree and type of phylogenetic differentiation. In relation to this criterion, criteria are given for the classification of the fundamental types of ontogenetic reproduction and for the classification of the types of ontogenetic generation cycles. Between the fundamental types of ontogenetic reproduction and the types of ontogenetic generation cycles there is a hierarchical relationship which shows that the former are components of the latter. Between the well-defined types of ontogenetic reproduction there exist many intermediate types.     

An enlarged European Union challenges priority settings in conservation

Biodiversity is declining worldwide under increasing human pressure. Since the location of and the threats are unevenly distributed and the resources available for conservation are limited, prioritization is essential to reduce the losses. Most conservation efforts until now proved to be ineffective in stopping the present worldwide decline of threatened species. We focus on the European Union (EU) after the repeated enlargements in the last decade, from 15 to 27 countries, by considering the present conservation priorities that have shifted towards a continental scale approach. The situation in the EU indicates that despite the differences in wealth across countries, there are no significant differences in the number and surface of protected areas between them, so re-evaluating conservation priorities at a continental scale and a reallocation of funds is required. A major limitation in priority settings for conservation is data availability. We recommend including in the decision process data provided by phylogeographic studies. This will prevent the decline of populations and species with evolutionary potential from centres of speciation and climate refugia. Recent EU members from central and eastern Europe still retain high biodiversity with a rather good conservation status. A large number of areas with high evolutionary potential identified by phylogeographic studies are located there and should be considered priorities within the context of global changes, as a proactive approach. We recommend a periodic re-evaluation of the status of species and habitats based on current research results, harmonization between the priority species listed in the conventions, directives and Red Lists at both EU and national levels.     

Rhizophagus irregularis MUCL 41,833 Association with Green Cuttings of Prunus sp. Rootstocks

Journal of Plant Growth Regulation - Sour cherry ‘Latvijas Zemais’ (Prunus cerasus) is a promising dwarfing rootstock for sweet cherries in Latvia, but low growing rate of newly...