Serological survey for potential disease agents of free-ranging cervids in six selected national parks from Germany

A total of 164 blood samples, collected from free-ranging red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) in six German national parks (NP) between 2000 and 2002, were assayed for antibodies against nine viral disease agents. Antibodies were only detected against the alpha-herpesviruses; specifically, bovine herpesvirus-1 (BHV-1) (22 of 157, 14%), cervid herpesvirus-1 (17 of 157, 10.8%), and caprine herpesvirus-1 (11 of 159, 6.9%). Titers ranged from 4 to 102. Most of the seropositive sera, and those with the highest antibody titers, were from red and roe deer in the Harz and Hochharz NP, which are connected and allow migration between the two. The distribution and specificity of antibodies detected in individual deer suggests that the three alpha-herpesviruses are circulating in these deer populations. No antibodies were detected against bovine viral diarrhea virus, epizootic hemorrhagic disease virus, bovine leukemia virus, bluetongue virus, foot-and-mouth disease virus, or sheep and goat poxvirus.     

Proteolytic Function of GPI-Anchored Plasma Membrane Protease Yps1p in the Yeast Vacuole and Golgi

Yps1p is a member of the GPI-anchored aspartic proteases which reside at the plasma membrane of Saccharomyces cerevisiae. Here we show that in Δerg6 cells, where a late biosynthetic step of the membrane lipid ergosterol is blocked, part of Yps1p was targeted to the vacuole. There it overtook proteolytic functions of the Pep4p protease, resulting in processing of pro-CPY to CPY in cells lacking the PEP4 gene. Yps1p was enriched in membrane microdomains, as it could be isolated in detergent-insoluble complexes from both normal and Δerg6 cells. Vacuolar Yps1 caused degradation of a mammalian sialyltransferase ectodomain fusion protein (ST6Ne), which was directed from the Golgi to the vacuole in both normal and Δerg6 cells. Unexpectedly, ST6Ne was degraded also when arrested in the Golgi in a temperature-sensitive sec7–1 mutant. Newly synthesized Yps1p, in transit to the plasma membrane, was also involved in the Golgi-associated degradation. These data show that GPI-anchored proteases, whose biological roles are unknown, may reside and function in different subcellular locations.     

Midsummer spatial variation in methane efflux from stands of littoral vegetation in a boreal meso-eutrophic lake

1. Spatial variation of methane (CH₄) efflux from the littoral zone of a meso‐eutrophic boreal lake was studied with a closed‐chamber technique for three summer days in 22 vegetation stands, consisting of three emergent and three floating‐leaved species. 2. Between‐species differences in CH₄ emission were significant. The highest emissions were measured from the emergent Phragmites australis stands (0.5–1.7 mmol m^?2 h^?1), followed by Schoenoplectus lacustris > Equisetum fluviatile > Nuphar lutea > Sparganium gramineum > Potamogeton natans. Within‐species differences between stands were not significant. 3. In P. australis stands, the stand‐specific mean CH₄ emission was significantly correlated with solar radiation, probably indicating the role of effective pressurised ventilation on CH₄ fluxes. The proportion of net primary production emitted as CH₄ was significantly higher in P. australis stands (7.4%) than in stands of S. lacustris and E. fluviatile (both 0.5%). 4. In N. lutea stands, CH₄ efflux was negatively correlated with the mean fetch and positively with the percentage cover of leaves on the water surface. There were no differences in CH₄ efflux between intact N. lutea leaves and those grazed by coleopteran Galerucella nymphaeae. In S. graminaeum and P. natans stands, CH₄ effluxes were not related to any of the measured environmental variables. 5. For all vegetation stands, the biomass above water level explained about 60% of the observed spatial variation in CH₄ emission, indicating the important role of plants as gas conduits and producers of substrates for methanogens in the anoxic sediment.     

Large-scale geographical variation confirms that climate change causes birds to lay earlier

Advances in the phenology of organisms are often attributed to climate change, but alternatively, may reflect a publication bias towards advances and may be caused by environmental factors unrelated to climate change. Both factors are investigated using the breeding dates of 25 long-term studied populations of Ficedula flycatchers across Europe. Trends in spring temperature varied markedly between study sites, and across populations the advancement of laying date was stronger in areas where the spring temperatures increased more, giving support to the theory that climate change causally affects breeding date advancement.     

Loss of Occludin and Functional Tight Junctions, but Not ZO-1, during Neural Tube Closure—Remodeling of the Neuroepithelium Prior to Neurogenesis

Neuroepithelial cells can generate nonepithelial cells, the neurons. Here we have investigated, for chick and mouse embryos, the epithelial character of neuroepithelial cells in the context of neurogenesis by examining the presence of molecular components of tight junctions during the transition from the neural plate to the neural tube. Immunoreactivity for occludin, a transmembrane protein specific to tight junctions, was detected at the apical end of the lateral membrane of neuroepithelial cells throughout the chick neural plate. During neural tube closure, occludin disappeared from all neuroepithelial cells. Correspondingly, the addition of horseradish peroxidase to the apical side of the neuroepithelium by injection into the amniotic cavity of mouse embryos revealed the presence of functional tight junctions in the neural plate (Embryonic Day 8), but not the neural tube (Embryonic Day 9). In contrast to occludin, expression of ZO-1, a peripheral membrane protein of tight junctions, increased from the neural plate to the neural tube stage, also being confined to the apical end of the lateral neuroepithelial cell membrane. This localization coincided with that of N-cadherin, whose expression increased concomitantly with the disappearance of occludin. We propose that the loss of tight junctions from neuroepithelial cells reflects an overall decrease in their epithelial nature, which precedes the generation of neurons.     

Species of Heterobasidion host a diverse pool of partitiviruses with global distribution and interspecies transmission

We investigated the geographic occurrence and genetic diversity of partitiviruses among 247 Heterobasidion specimens representing seven species and originating from Europe, Asia, and North America. Based on sequence analysis, partitiviruses were relatively rare, and occurred only in about 5 % of the Heterobasidion isolates analyzed, constituting a minority (about 28 %) of all virus-infected [double-stranded RNA (dsRNA)-positive] isolates. Altogether ten virus strains were characterized in sequence: one complete genome sequence of 3893 bp, six complete RNA-dependent RNA polymerase sequences of 2000-2033 bp, and three partial polymerase sequences. Based on phylogenetic analysis, the virus strains were assigned into three putative partitivirus species: HetRV1 (Heterobasidion RNA virus 1), HetRV4, and HetRV5. Degenerate consensus primers were designed for RT-PCR detection of these virus species. HetRV1 occurred in five different Heterobasidion species, and resembled the previously described Heterobasidion annosum virus (HaV). Highly similar HetRV1 strains with 98 % nucleotide level similarity were found from H. parviporum (member of the H. annosum species complex) and H. australe (member of the H. insulare complex) growing in the same region in Bhutan. This observation suggests recent virus transmission between these taxonomically distant Heterobasidion species in nature. It was also shown that HetRV1 can be transmitted by mycelial contact between the H. annosum and H. insulare complexes. The two other virus species, HetRV4 and HetRV5, were closely related to the Amasya Cherry Disease-associated mycovirus, to Heterobasidion parviporum partitivirus Fr110B, and also to several plant-infecting alphacryptoviruses. These results are in accordance with the view of a close evolutionary relationship between partitiviruses of plants and fungi.     

Origin of symmetrical triradial chromosomes in human cells

We have collected 23 sporadic symmetrical triradial chromosomes (plus one D with duplicate satellites), 22 from cultured lymphocytes and one from a bone marrow cell. Fifteen triradials were from patients with Bloom's syndrome, and two from a Fanconi's anemia patient. The following chromosomes and chromosome groups were involved: 1, 2, 3, 4, 5, C (11 identified), D, and 17. The branchpoints were localized nonrandomly. Regions in or near centric heterochromatin were often involved. Some of the branchpoints are regions which also contain a high number of mitotic chiasmata. When the present sporadic triradials combined with those from the literature were compared with triradials with branchpoints in the fragile regions, the localized branchpoints were different in these two groups. Our conclusion that most — possibly all — symmetrical triradials are caused by partial endoreduplication is based on the following observations: the shape of the triradials which shows that the extra segments are paired with their intact sister chromatids and not with each other; the failure of X-rays in G₂ to increase the incidence of symmetrical triradials; the fact that in some cases the end of the extra segment is joined to its intact sister chromatid; and the occurrence of duplicate satellites.     

Endomitosis in human trophoblast

Summary Endopolyploidy, which arises through the duplication of DNA without accompanying nuclear division, occurs in large numbers of lower and higher plants and animals, including the best known, the salivary gland nuclei of Drosophila. Endomitosis is one of the processes leading to endopolyploidy, in which the stages of mitosis (prophase, metaphase, anaphase) take place inside the nuclear membrane and without spindle formation. In mammals, endomitosis has been observed in the trophoblast of the placenta of the mouse, rat and rabbit. This is the first report of endomitosis in a normal human tissue, the trophoblast of first trimester human placenta.This research was supported in part by the Foundation for Reproductive Research and Education, Inc., Northwestern University, Department of Obstetrics and Gynecology.     

Mitotic crossing-over and segregation in man

Summary Although clear genetic evidence of mitotic crossing-over is lacking in man, observations of mitotic chiasmata in normal cells (0.1–1 per 1000) and in Bloom's syndrome (BS) cells (5–150 per 1000) demonstrate its occurrence. That mitotic chiasmata are true exchanges is concluded from the occurrence of heteromorphic bivalents and the pattern of sister chromatid exchanges in mitotic bivalents. Several observations demonstrate that chiasmata are different in principle from chromatid translocations which simply happen to take place at homologous loci. For example, the ratio of adjacent exchanges to mitotic chiasmata is 1/20–1/60, whereas this ratio is approximately 1:1 for chromatid translocations. Furthermore, mitotic chiasmata make up a very high proportion of total quadriradials (QRs): 48% in normal untreated cells and 90% in BS cells.Close proximity of homologous chromosomes promotes mitotic crossing-over. Thus in normal diplochromosomes, the incidence is increased a hundred-fold as compared to diploid cells. However, closeness of homologues is not the only factor promoting crossing-over; the BS gene specifically promotes exchanges between homologous segments as shown by the roughly 15-fold increase of chiasmata in BS diplochromosomes as compared to normal diplochromosomes.Mitotic chiasmata are distributed extremely nonrandomly in different chromosomes and chromosome segments. The preferred sites are short Q-dark regions, 3p21, 6p21, 11q13, 12q13, 17q12, and 19p13 or q13 being veritable hot spots. Our preferred hypothesis is that the hot spots have higher gene densities than other regions. Consequently they are active and extended in interphase which would promote their pairing and chiasma formation.Segregation after mitotic corssing-over in satellite stalks can be demonstrated by means of distinct satellites. In a BS patient there were 31 different patterns for Q-bright satellites in 58 cells. Segregation after presumed crossing-over has also been seen in three dicentric chromosomes with one centromere inactivated. Recombination in satellite stalks in BS resulted in 12/58 cells homozygous for Q-bright satellites. In two of these cells, two chromosomes were homozygous for Q-bright satellites, and in one cell, three chromosomes were homozygous. This high degree of homozygosity which obviously applies to other chromosome regions too, may explain the high incidence of malignant disease in BS on the assumption that cancer is caused by recessive genes.     

Mitotic recombination and segregation of satellites in Bloom's syndrome

Mitotic recombination in satellite stalks — a phenomenon often difficult to distinguish from satellite association — was studied in a sister and a brother with Bloom's syndrome. Segregation after recombination was analyzed in the lymphocytes of the sister who had Q-bright satellites. Her cells varied greatly both in regard to the acrocentrics which displayed Q-bright satellites and the number of such satellites per cell. In 58 cells a total of 31 different patterns were seen. In 83 cells of 6 controls who also had Q-bright satellites on at least one acrocentric chromosome, not one cell was found in which the pattern differed from that characteristic of the person. Obviously exchanges between satellite stalks in patients with Bloom's syndrome are fairly frequent (estimated lower limit 6/1000) and very rare in persons who do not have this syndrome (estimated 0.1/1000).