An initial investigation of the taxonomic status of Saussurea esthonica Baer ex Rupr. utilising DNA markers and sequencing

Saussurea esthonica Baer ex Rupr. is a species of the genus Saussurea that is found in Estonia, Latvia and the Leningrad region in Russia. Only two populations have been identified in Latvia, and it is protected at the national level. The taxonomic relationships within this genus are unclear, and the majority of phylogenetic studies of this genus have been undertaken on species found in Asia. The relationship of S. esthonica to the other Saussurea species is equally unclear, with some suggestions that this species should be regarded as a subspecies of S. alpina. Two different molecular marker techniques, iPBS and ITS sequencing, were utilised to investigate the phylogeny of S. esthonica and its relationship with other Saussurea species, including three European species: S. alpina, S. pygmaea and S. discolor. The phylogeny obtained with the iPBS markers was similar to that obtained with the ITS sequences. The phylogenies obtained were broadly consistent with previous morphological classification, with S. alpina, S. esthonica and S. discolor clustering together. S. pulchella, S. albescens and S. candicans also formed one discrete cluster. The taxonomic status of S. esthonica was not unambiguously determined, with Norwegian and Italian S. alpina populations clustering with Latvian and Estonian S. esthonica populations, and the Austrian S. alpina populations clustering with Austrian S. discolor populations.     

Microdosimetric Study for Nanosecond Pulsed Electric Fields on a Cell Circuit Model with Nucleus

Recently, scientific interest in electric pulses, always more intense and shorter and able to induce biological effects on both plasma and nuclear membranes, has greatly increased. Hence, microdosimetric models that include internal organelles like the nucleus have assumed increasing importance. In this work, a circuit model of the cell including the nucleus is proposed, which accounts for the dielectric dispersion of all cell compartments. The setup of the dielectric model of the nucleus is of fundamental importance in determining the transmembrane potential (TMP) induced on the nuclear membrane; here, this is demonstrated by comparing results for three different sets of nuclear dielectric properties present in the literature. The results have been compared, even including or disregarding the dielectric dispersion of the nucleus. The main differences have been found when using pulses shorter than 10 ns. This is due to the fact that the high spectral components of the shortest pulses are differently taken into account by the nuclear membrane transfer functions computed with and without nuclear dielectric dispersion. The shortest pulses are also the most effective in porating the intracellular structures, as confirmed by the time courses of the TMP calculated across the plasma and nuclear membranes. We show how dispersive nucleus models are unavoidable when dealing with pulses shorter than 10 ns because of the large spectral contents arriving above 100 MHz, i.e., over the typical relaxation frequencies of the dipolar mechanism of the molecules constituting the nuclear membrane and the subcellular cell compartments.     

Kin cohesiveness and possible inbreeding in the mouthbrooding tilapia Sarotherodon melanotheron (Pisces Cichlidae)

Four microsatellite markers were used to study genetic variation among individuals of the mouthbrooding tilapia Sarotherodon melanotheron (Rüppel 1852) caught in separate but adjacent shoals. A comparison was also made with fish from six other localities. Populations originating from riverine environments appear to be panmictic, while samples from open waters such as lagoons showed highly significant heterozygote deficiencies. For instance, at the 33-allele locus SMEL4, 32 homozygous individuals were observed among the 82 individuals from the same lagoon location instead of only five homozygotes expected if random mating occurred. A further assessment of the genetic similarity of individuals within each shoal, validated by robust permutation techniques requiring no precise knowledge of gene frequencies, showed that related individuals tend to aggregate, and suggested that mating occurs preferentially within small groups of kin. Cichlids are often presented as a group of fish where microallopatric speciation takes place. The possible link between kin aggregation, inbreeding and shoaling behaviour we propose here may have important consequences for our understanding of the mechanisms involved in this fast speciation process.     

Induction of CAD gene amplification by restriction endonucleases in V79,B7 Chinese hamster cells

The restriction endonucleases PvuII, BamHI and EcoRI were tested for their ability to induce gene amplification in V79,B7 Chinese hamster cells. The results indicate that treatment with these enzymes efficiently increases the frequency of clones resistant to N-phosphonacetyl-L-aspartate, indicating induction of CAD gene amplification.     

Micro-habitat drivers of saproxylic beetle assemblages in old woodlands of Mediterranean cork oak (Quercus suber)

1. Cork oak landscapes are fascinating ecosystems, historically managed for cork extraction. The persistence in this habitat of many hollow veteran trees provides suitable micro-habitats for saproxylic beetles.
2. We investigated the saproxylic beetle community of two isolated cork oak woodlands of central Italy with different degree of recovery after human transformation: (1) an open woodland and (2) a dense mixed woodland, both dominated by cork oak trees.
3. We found endemic, rare and threatened saproxylic beetles in both the areas, confirming the important conservation value of cork oak landscapes. In the open woodland we observed a higher number of species in all trophic categories, except for mycophagous specialists. Several microhabitat variables reflected the different stage of recovery of the two woodlands.
4. Our findings suggest the crucial role of diversified environments in protected areas: even a small difference in the degree of recovery (i.e., tree closeness) can affect the number of beetle species. Specifically, we found (1) more xerophilous species in the open woodland and (2) more mesophilous species in the dense mixed woodland.

     

Cytogenetic manifestations associated with the reversion, by gene amplification, at the HGPRT locus in V79 Chinese hamster cells

Some HGPRT spontaneous revertants were isolated from a mutant line (E2) of V79 Chinese hamster cells and phenotypically characterized. Dot-Blot hybridization with a 32P-labelled HGPRT probe revealed an increase in the number of HGPRT sequences in some of these revertants, suggesting the occurrence of gene amplification. Cytogenetic analysis performed in three of these revertants showed a characteristic abnormally banding region (ABR) on the elongated p arm of the X chromosome. In situ hybridization in one revertant (RHE2) showed that the amplified sequences reside on the p+ arm of the X chromosome in two different localizations. Because of the very probably clonal origin of the revertant, these features indicate that the amplified sequences might rearrange after their integration into the chromosome.