DNA-based prenatal diagnosis of the infantile form of neuronal ceroid lipofuscinosis (INCL,CLN1)

Eleven fetuses at risk for the infantile form of neuronal ceroid lipofuscinosis (INCL, CLN1) were studied using DNA markers and the results were compared with the results of electron microscopy (EM) of chorionic villus specimens from pregnancies in the first or early second trimester of pregnancy. In four cases, the prenatal diagnosis was made independently with both methods, and in seven cases, the EM diagnosis was confirmed postnatally or from autopsy material using RFLP analysis. The two methods gave concordant results in all cases. The DNA analysis based on RFLP haplotypes also for the first time facilitates reliable carrier diagnostics. RFLP analysis based on polymorphic markers closely linked to the INCL locus is now available for prenatal diagnosis of this fatal brain disease, whose biochemical background is totally unknown and for which no treatment is available.     

Chromosomal prenatal diagnosis: Study of 936 cases of intrauterine abnormalities after ultrasound assessment

Nine hundred and thirty-six prenatal chromosomal analyses were performed by four cytogenetic centres after ultrasound diagnosis of fetal abnormalities, amniotic fluid disorders, fetal growth retardation, and fetal or placental abnormalities. During the same period, 6515 fetal karyotypes were analysed because of maternal age. Frequencies of chromosomal aberrations in each case were respectively 4·4, 6·7 and 15·8 per cent, compared with 3·18 per cent when the fetal karyotype was performed because of maternal age. High rates of chromosomal aberrations are observed in cases of cervical hygroma, limb abnormalities, omphaloceles, duodenal stenosis, hydrocephalus, and facial abnormalities. In the case of polymalformations, this rate was 29·2 per cent. When malformations were seen together with an amniotic fluid disorder or growth retardation, 21·5 per cent chromosomal aberrations were observed. This frequency was 10·4 per cent when growth retardation was associated with an amniotic fluid disorder. Trisomy 13, 18, 21 and monosomy X accounted for 4/5 of all abnormalities in which we observed a high rate of triploidies (4·9 per cent) and balanced (3·3 per cent) or unbalanced (9·8 per cent) non-Robertsonian structural abnormalities. Sonographic ascertainment of these aberrations and prenatal characteristics of major anomalies are discussed.     

Female tree swallows (<Emphasis Type="Italic">Tachycineta bicolor</Emphasis>) increase offspring heterozygosity through extrapair mating

Recent attention has focused on genetic compatibility as an adaptive function for why females engage in extrapair mating. We tested the genetic compatibility hypothesis in tree swallows (Tachycineta bicolor) over five breeding seasons using data from ten microsatellite loci. Tree swallows are socially monogamous passerines exhibiting high levels of extrapair paternity. Overall, we found that 47% of offspring were the result of extrapair fertilizations, and 83% of females produced at least one extrapair offspring. Consistently for all years, extrapair offspring were more heterozygous than their maternal half-siblings, which is in accordance with the genetic compatibility hypothesis. The difference was largely caused by the high heterozygosity of extrapair offspring sired by unknown males, suggesting that females are engaging in extrapair copulations with geographically distant males to increase the likelihood of being inseminated by a more compatible mate. Our findings support the idea that postcopulatory mechanisms are important for females when assessing potential sires for their offspring.     

多环芳烃对海洋贝类多生物水平毒性效应的研究进展

多环芳烃类化合物(PAHs)是海洋中常见的一类持久性有机污染物,对海洋生态安全及海洋生物健康造成严重威胁。海洋贝类作为海洋生态毒理学研究的模式生物,其滤食性、固着性等生理特点使其对PAHs具有较高的生物蓄积能力,可以在不同生物水平产生一系列的毒性效应。本文综述目前PAHs在海洋贝类多种生物水平所造成的生物毒性效应及其检测方法的研究进展,重点从个体生理特征、组织结构、细胞毒性和基因毒性4个层次展开讨论,为更有效地利用海洋贝类这一模型生物,深入开展PAHs对海洋生物的致毒效应与机制研究提供思路与检测方法参考。     

Effects of skin and hydraulic fractures on SVE wells

Soil vapor extraction (SVE) systems are intended to cause substantial volumes of air to flow through the subsurface with the purpose of removing volatile contaminants. The effectiveness of SVE can be influenced by any effect that changes the specific gas capacity (discharge as a function of vacuum) of a well. Skins of low permeability material enveloping a well bore are widely recognized to affect the performance of wells used to recover water, natural gas, or petroleum, and skin can also significantly diminish the performance of an SVE well. Skins a few mm thick consisting of material whose gas phase permeability is 0.01 of the formation can reduce the specific gas capacity of an SVE well by factors of 2 to 10 or more. Hydraulic fractures created in the vicinities of shallow wells commonly resemble sand-filled layers shaped like flat-lying disks or gently dipping saucers. The contrast between the gas-phase permeability of the sand in the fracture and that of the formation is particularly important, with significant effects requiring the ratio to be greater than approximately 50. Shallow hydraulic fractures filled with several tenths of m3 of sand in formations that are several orders of magnitude less permeable than that of the enveloping formation should increase specific gas capacity by factors of 10 or more. Field tests of the effects of hydraulic fractures on the performance of SVE were conducted by creating four wells intersecting fractures and a suite of control wells created using conventional methods in silty saprolite. Specific gas capacities ranged over more than an order of magnitude for 10 wells completed within a small area (2 m2) and at the same depth. Specific capacities correlate to the drilling method that was used to create the bore for the well: lowest values occurred in wells drilled with a machine auger, slightly better results were obtained using a Shelby tube, and the best results were obtained from conventional wells bored with a hand auger. Skin factors determined for wells created with a machine auger could be explained by a layer 1 cm thick that has 0.007 times the permeability of the enveloping material, which could readily have been created during the drilling procedure. Specific capacities of wells intersecting hydraulic fractures were 5 to 100 times more than those of conventional wells. The large difference in performance appears to be due in part to the beneficial effects of the fracture, and in part to the detrimental effects of well skin.     

Time course of isocyanate emission from curing polyurethane adhesives

The time course of isocyanate emission from curing polyurethane (PUR) resins and adhesives was studied in two different emission test chambers. The measured emissions were strongly dependent on the type of experiment. The adhesives under investigation contained different types of diisocyanates and are used for different applications, e.g. for fixing of textile floor coverings. The influence of the curing mechanism on emission was studied by comparing the emission curves of one-component adhesives (OCA) and two-component adhesives (TCA). For TCA, the decrease in isocyanate emission was found to follow a two-step process during curing. In the first step, the emission is dominated by surface evaporation, and the decay of emission is mainly caused by the decrease in monomer content due to reaction. In the second step, the release is limited by internal diffusion. The influence of monomer reactivity on the emission profile could be demonstrated for 2,4′- and 4,4′-MDI. The less-reactive 2,4′-MDI caused prolonged emission. A strong dependence of emission rates on temperature and adhesive viscosity was also obvious. The evaluation of emission rates of different commercially available PUR adhesives showed the highest emission from systems that are applied at high temperatures. The high reactivity of diisocyanates requires special techniques for sampling and analysis. Therefore, an analytical method using HPLC–MS/MS was developed that enables limits of quantitation of <5 ng/m³ with a sampling volume of 100 l.     

Small testes size despite high extra-pair paternity in the pair-living nocturnal primate Phaner furcifer

Sperm competition will be the inevitable consequence of polyandrous mating behavior if two or more males inseminate a single female. It has been demonstrated for a wide variety of animals that males adapt to this situation behaviorally, physiologically and morphologically, e.g. by evolving relatively large testes size to produce more sperm. All pair-living primates investigated so far were found to have relatively small testes, suggesting a monandrous mating system. We investigated the relationship between extra-pair paternity (EPP) rate as a measure of sperm competition intensity and relative testes size in a pair-living primate, the fork-marked lemur (Phaner furcifer). Paternity exclusion analyses for seven offspring using six polymorphic DNA-microsatellite markers suggested a high EPP rate. Female nocturnal travel distances were longer during the mating season, suggesting that females take an active role in achieving extra-pair copulations (EPCs). Surprisingly, fork-marked lemur testes size was relatively small compared to 23 other lemuroid primates, a result that is in contrast to predictions of sperm competition theory. Neither possible behavioral and morphological adaptations to an alternative paternity guard (i.e. mate guarding), nor sampling biases, phylogenetic constraints, and population density effects explain the absence of large testes in a species with high EPP, a phenomenon also known from birds with moderate to low EPP rates. We conclude that more data are needed on the frequency of EPCs, the timing of in-pair and extra-pair copulations, as well as the role of female choice, to explain why males of some species apparently do not adapt to sperm competition.Communicated by S. Alberts     

Manipulation of nest-box density affects extra-pair paternity in a population of blue tits (<Emphasis Type="Italic">Parus caeruleus</Emphasis>)

We tested the effect of manipulation of breeding density on the occurrence of extra-pair paternity in a blue tit (Parus caeruleus) population during two consecutive years. In a homogeneous oak forest, nest-box manipulation provided a high density plot (plot A, 1.10 and 1.32 pairs/ha) and a low density plot (plot B, 0.43 and 0.46 pairs/ha). Microsatellite analysis on 91 broods revealed a higher proportion of extra-pair paternity in broods in plot A (mean of 17.2%) than in plot B (mean of 11.4%). A correlative approach showed that the proportion of extra-pair young in broods was affected by the number of breeding neighbours within 100 m around the nest-box, by the distance to the nearest breeding neighbour and by an additional plot effect. However, the nearest neighbours accounted for only 39.3% of extra-pair paternities and distance to extra-pair fathers was significantly higher than the nearest neighbour distance in both plots. This implies that the effect of density on the occurrence of extra-pair paternities is associated with active female choice to enhance the brood fitness. Although there were more extra-pair young in broods when density was high, the number of extra-pair fathers did not increase and stayed close to one. We suggest that density increases the cost of mate guarding by males, thereby increasing the possibility for females to solicit extra-pair paternities to the cuckolding male they have chosen. Finally, we discuss why correlatives approaches do not always show evidence for an effect of breeding density on extra-pair paternity occurrence.Communicated by M. Soler     

Paternity and performance of golden-winged warblers (<Emphasis Type="Italic">Vermivora chrysoptera</Emphasis>) and golden-winged X blue-winged warbler (<Emphasis Type="Italic">V. pinus</Emphasis>) hybrids at the leading edge of a hybrid zone

We examined two components of reproduction in a population of golden-winged warblers in the initial stages of hybridization with blue-winged warblers. First, we used genetic analyses of mate choice to determine whether copulations outside the social pairbond (extra-pair copulations; EPCs) occur in these species and their hybrids. Second, we compared several aspects of reproduction (pairing success, clutch size, hatching success, fledging success, and loss of paternity) in nests raised by phenotypic golden-winged warblers to those raised by hybrid individuals. Together, these data provide us with the first quantitative analysis of reproduction within this hybrid system. Our data suggest several reasons why the level of hybridization between these species is likely being underestimated. First, many birds in our population showed only subtle phenotypic signs of introgression indicating that hybrid status can only be determined by close examination. Second, high rates of extra-pair paternity indicate that we cannot base our estimates of hybridization on pairbond data alone. More than 30% of nestlings were the result of EPCs, occurring in 55% of all nests. Third, there was no difference in the number of hybrid or “pure” golden-winged warbler males chosen as social mates, relative to their abundance. Indeed, based on several components of nesting success, it appears as though hybrids are having as great realized reproductive success as are phenotypic golden-winged warblers in our population. Accordingly, we argue that hybrid reproductive success and extra-pair fertilizations are likely playing a major role in the ongoing hybridization between blue-winged and golden-winged warblers. Studies such as this one are important to our understanding of the process and implications of hybridization, and findings may be applicable to other hybrid zones.     

Sperm competition in horseshoe crabs (Limulus polyphemus)

Male horseshoe crabs have two mating tactics. Some males come ashore attached to a female (clasping the posterior margin of the females' carapace with their modified pedipalps) and nest with her on the intertidal portion of the beach during the high tide. Other males come ashore unattached and crowd around nesting couples. Fertilization is external and unattached males that are in contact with a pair, i.e. satellite males, release sperm, so the assumption has been that they are fertilizing eggs. We conducted a paternity analysis to determine the proportion of eggs fertilized by attached and satellite males. Pairs with one satellite were observed during nesting on beaches in Florida and Delaware and their eggs were collected and reared to the late trilobite or first instar horseshoe crab stage. DNA was extracted from these offspring and from each adult (female, attached and satellite male) for use in paternity analysis. A Limulus-specific hypervariable microsatellite locus was identified and primers were constructed to amplify this locus via the polymerase chain reaction (PCR). Genotypes of putative parents and offspring were determined by resolving length variants of these PCR products on acrylamide gels. This allowed us to determine parentage of the offspring. We demonstrate that satellite males fertilized 40% of the eggs on average, attached males fertilized 51% and 4% of the eggs that were laid by the female were fathered neither by the attached male nor by the satellite (and 5% could not be determined unambiguously). There is high variability in the success of satellite males, ranging from 0 to 88%. Part of this variability can be explained by the position of the satellite relative to the attached male. We discuss the mechanics of fertilization and the possible advantages for multiple mating in this species.