Pattern and frequency of nondisjunction in oocytes from the Djungarian hamster are determined by the stage of first meiotic spindle inhibition

In order to study the mechanisms of nondisjunction at meiosis I in oocytes gonadotropin-stimulated Djungarian hamsters were treated at two stages [4.5 and 6 h post human chorionic gonadotropin (HCG)] during the preovulatory period with 1000 mg/kg Carbendazim (MBC). The compound, known to bind fast but reversibly to mammalian tubulin, was chosen to investigate whether the stage at which spindle function is inhibited affects the pattern of nondisjunction. Ovulated oocytes were cytologically prepared and scored for hyperhaploidy, diploidy and presegregation. Application at an early spindle phase, 4.5 h post HCG, to females stimulated with a low gonadotropin dose [3 IU pregnant mares serum (PMS); 2 IU HCG] caused a high frequency of nondisjunction (40.6%) with a more or less nonspecific pattern of malsegregated bivalents. Treatment at a late stage of spindle function (6 h post HCG) resulted in a less frequent (22.5%) but highly preferential malsegregation of those A-D group bivalents thought earlier to be late segregators. On the other hand, oocytes from females primed with a high (10 IU PMS and HCG) gonadotropin dose, a treatment assumed to delay meiosis by approximately 1.5 h, responded to MBC treatment at the late stage (6 h) with a nonspecific pattern and a high frequency (71.2%) of nondisjunction. The latter result is comparable to that in which MBC was given at the early stage (4.5 h) and after a low gonadotropin dose. The high nondisjunction response additionally indicates that spindles in hypergonadotropic stimulated oocytes are more susceptible and/or that the concentration of the inhibitor is higher in such oocytes. Only few oocytes with presegregation (3.1%; 0.0%; 1.7%) and few diploid oocytes (3.3%; 1.5%; 3.2%) with complete inhibition of meiosis I were observed. We conclude, that in Djungarian hamsters (1) the segregation of bivalents at meiosis I is asynchronous with the large A-D bivalents segregating last, (2) the phase in which spindle function is inhibited determines the pattern of nondisjunction, and (3) the resumption of meiosis I — from dictyotene to metaphase II — does not follow a rigidly timed programme but depends on the conditions of follicular maturation.     

PASSAGE OF A MARINE BROWN ALGAL DNA VIRUS FROM ECTOCARPUS FASCICULATUS (ECTOCARPALES,PHAEOPHYCEAE) TO MYRIOTRICHIA CLAVAEFORMIS (DICTYOSIPHONALES,PHAEOPHYCEAE): INFECTION SYMPTOMS AND RECOVERY1

A dsDNA virus (EfasV-1) isolated from Ectocarpus fasciculatus Harvey infected Myriotrichia clavaeformis Harvey, a species belonging to a different brown algal order. The virus did not complete its infection cycle in the foreign host but caused infertility due to malformed reproductive structures. After some time in culture, the host's reproductive capacity was sometimes restored with concomitant loss of at least part of the viral genome. This incidence of interordinal virus transfer is discussed in relation to possibilities for virus-mediated horizontal gene transfer in brown algae.     

The dcuD (former yhcL) gene product of Escherichia coli as a member of the DcuC family of C4-dicarboxylate carriers: lack of evident expression

The dcuD gene (formerly yhcL) of Escherichia coli shows significant sequence similarity only to the dcuC gene of E. coli, which encodes a C4-dicarboxylate carrier (DcuC) that functions during anaerobic growth. Inactivation of dcuD had no effect on the growth of E. coli under a large number of conditions and led to no detectable changes in phenotype. Translational dcuD′-′lacZ gene fusions were not significantly expressed in the presence of dicarboxylates or monocarboxylates under oxic or anoxic conditions. Other potential substrates such as amino sugar derivatives, amino acids, and α-aspartyl dipeptides also did not lead to expression of dcuD. Changes in medium composition, pH, ionic strength, and temperature had no significant effects on dcuD expression. A dcuD gene amplified from a natural isolate of E. coli was not expressed in wild-type and E. coli K-12 backgrounds. Cloning of dcuD behind an inducible promoter resulted in the synthesis of a protein of the expected size (49 kDa), which, however, did not complement for the loss of DcuC or other C4-dicarboxylate carriers. It is suggested that dcuD encodes a protein of the DcuC family of anaerobic C4-dicarboxylate carriers and that dcuD is not significantly expressed or is expressed only under conditions not related to carboxylate metabolism. When two adjacent open reading frames (y0585 and y0586) from Haemophilus influenzae are fused, the resulting hypothetical protein has sequence similarity to DcuC and DcuD. Received: 11 May 1999 / Accepted: 6 July 1999     

Isolation of Three Novel Rat and Mouse Papillomaviruses and Their Genomic Characterization

Despite a growing knowledge about the biological diversity of papillomaviruses (PV), only little is known about non-human PV in general and about PV mice models in particular. We cloned and sequenced the complete genomes of two novel PV types from the Norway rat (Rattus norvegicus; RnPV2) and the wood mouse (Apodemus sylvaticus; AsPV1) as well as a novel variant of the recently described MmuPV1 (originally designated as MusPV) from a house mouse (Mus musculus; MmuPV1 variant). In addition, we conducted phylogenetic analyses using a systematically representative set of 79 PV types, including the novel sequences. As inferred from concatenated amino acid sequences of six proteins, MmuPV1 variant and AsPV1 nested within the Beta+Xi-PV super taxon as members of the Pi-PV. RnPV2 is a member of the Iota-PV that has a distant phylogenetic position from Pi-PV. The phylogenetic results support a complex scenario of PV diversification driven by different evolutionary forces including co-divergence with hosts and adaptive radiations to new environments. PV types particularly isolated from mice and rats are the basis for new animal models, which are valuable to study PV induced tumors and new treatment options.     

Habitat selection and population trends in terrestrial bird species of Robinson Crusoe Island: habitat generalists versus forest specialists

Habitat loss and degradation on oceanic islands are key processes leading to population decline of endemic birds and facilitating the establishment of invasive bird species. In this study, carried out in the Robinson Crusoe Island, we assessed density and habitat selection of terrestrial bird species, including juan fernandez firecrown and juan fernandez tit-tyrant, two endemics, as well as green-backed firecrown and austral thrush, which apparently originate from the mainland. Results show that perturbed habitats contained a low density of the endemic species whereas the mainland species were significantly more abundant in perturbed scrub habitats. Bird species show different habitat selection patterns, with endemics selecting for native forest and mainland species selecting for perturbed habitats, or using them at random. Bird species experienced temporal trends in their overall population sizes, with the endemic tit-tyrant suffering a significant decline in its population size of about 63% between 1994 and 2009. Only mainland species exhibited temporal changes in habitat use, significantly reducing their densities in the preferred scrub habitats, possibly as a response to decreased habitat quality. Thrushes apparently were able to compensate the population decrease in one non native habitat type by using native forests, a habitat giving them the opportunity of preying on nests of endemic species. We conclude that endemic bird species behave as specialists whereas the mainland species must be treated as invasive generalists on Robinson Crusoe Island.     

Effects of Exhaustive Aerobic Exercise on Tryptophan-Kynurenine Metabolism in Trained Athletes

Exhaustive exercise can cause a transient depression of immune function. Data indicate significant effects of immune activation cascades on the biochemistry of monoamines and amino acids such as tryptophan. Tryptophan can be metabolized through different pathways, a major route being the kynurenine pathway, which is often systemically up-regulated when the immune response is activated. The present study was undertaken to examine the effect of exhaustive aerobic exercise on biomarkers of immune activation and tryptophan metabolism in trained athletes. After a standardized breakfast 2 h prior to exercise, 33 trained athletes (17 women, 16 men) performed an incremental cycle ergometer exercise test at 60 rpm until exhaustion. After a 20 min rest phase, the participants performed a 20 min maximal time-trial on a cycle ergometer (RBM Cyclus 2, Germany). During the test, cyclists were strongly encouraged to choose a maximal pedalling rate that could be maintained for the respective test duration. Serum concentrations of amino acids tryptophan, kynurenine, phenylalanine, and tyrosine were determined by HPLC and immune system biomarker neopterin by ELISA at rest and immediately post exercise. Intense exercise was associated with a strong increase in neopterin concentrations (p<0.001), indicating increased immune activation following intense exercise. Exhaustive exercise significantly reduced tryptophan concentrations by 12% (p<0.001) and increased kynurenine levels by 6% (p = 0.022). Also phenylalanine to tyrosine ratios were lower after exercise as compared with baseline (p<0.001). The kynurenine to tryptophan ratio correlated with neopterin (r = 0.560, p<0.01). Thus, increased tryptophan catabolism by indoleamine 2,3-dioxygenase appears likely. Peak oxygen uptake correlated with baseline tryptophan and kynurenine concentrations (r = 0.562 and r = 0.511, respectively, both p<0.01). Findings demonstrate that exhaustive aerobic exercise is associated with increased immune activation and alterations in monoamine metabolism in trained athletes which may play a role in the regulation of mood and cognitive processes.     

New records of benthic marine algae and Cyanobacteria for Costa Rica,and a comparison with other Central American countries

We present the results of an intensive sampling program carried out from 2000 to 2007 along both coasts of Costa Rica, Central America. The presence of 44 species of benthic marine algae is reported for the first time for Costa Rica. Most of the new records are Rhodophyta (27 spp.), followed by Chlorophyta (15 spp.), and Heterokontophyta, Phaeophycea (2 spp.). Overall, the currently known marine flora of Costa Rica is comprised of 446 benthic marine algae and 24 Cyanobacteria. This species number is an under estimation, and will increase when species of benthic marine algae from taxonomic groups where only limited information is available (e.g., microfilamentous benthic marine algae, Cyanobacteria) are included. The Caribbean coast harbors considerably more benthic marine algae (318 spp.) than the Pacific coast (190 spp.); such a trend has been observed in all neighboring countries. Compared to other Central American countries, Costa Rica has the highest number of reported benthic marine algae; however, Panama may have a similarly high diversity after unpublished results from a Rhodophyta survey (Wysor, unpublished) are included. Sixty-two species have been found along both the Pacific and Caribbean coasts of Costa Rica; we discuss this result in relation to the emergence of the Central American Isthmus.     

A Novel Malaria Vaccine Candidate Antigen Expressed in Tetrahymena thermophila

Development of effective malaria vaccines is hampered by the problem of producing correctly folded Plasmodium proteins for use as vaccine components. We have investigated the use of a novel ciliate expression system, Tetrahymena thermophila, as a P. falciparum vaccine antigen platform. A synthetic vaccine antigen composed of N-terminal and C-terminal regions of merozoite surface protein-1 (MSP-1) was expressed in Tetrahymena thermophila. The recombinant antigen was secreted into the culture medium and purified by monoclonal antibody (mAb) affinity chromatography. The vaccine was immunogenic in MF1 mice, eliciting high antibody titers against both N- and C-terminal components. Sera from immunized animals reacted strongly with P. falciparum parasites from three antigenically different strains by immunofluorescence assays, confirming that the antibodies produced are able to recognize parasite antigens in their native form. Epitope mapping of serum reactivity with a peptide library derived from all three MSP-1 Block 2 serotypes confirmed that the MSP-1 Block 2 hybrid component of the vaccine had effectively targeted all three serotypes of this polymorphic region of MSP-1. This study has successfully demonstrated the use of Tetrahymena thermophila as a recombinant protein expression platform for the production of malaria vaccine antigens.