Conserved sequences and cell-specific DNase I hypersensitive sites upstream from the co-ordinately expressed alpha I- and alpha II-globin genes of Xenopus laevis

The globin gene family of Xenopus laevis comprises pairs of closely related genes that are arranged in two clusters, each pair of genes being co-ordinately and stage-specifically expressed. To get information on putative regulatory elements, we compared the DNA sequences and the chromatin conformation 5' to the co-ordinately expressed adult alpha-globin genes. Sequence analysis revealed a relatively conserved region from the cap site up to position -289, and further upstream seven distinct boxes of homology, separated by more diverged sequences or deletions/insertions. The homology boxes comprise 22 to 194 base-pairs showing 78 to 95% homology. Analysis of chromatin conformation showed that DNase I preferentially cuts the upstream region of both genes at similar positions, 5' to the T-A-T-A and the C-C-A-A-T boxes, only in chromatin of adult erythroblasts and erythrocytes, where adult globin genes are expressed, but not in chromatin of adult liver cells or larval erythrocytes, where these genes are silent. This suggests that cell- and stage-specific activation of these genes coincides with specific changes in chromatin conformation within the proximal upstream region. No difference was found in the nucleotide sequence within the DNase I hypersensitive region proximal to the adult alpha 1-globin gene in DNA from embryonic cells, in which this gene is inactive, and adult erythrocytes, expressing this gene.

Trends of mean and extreme temperature indices since 1874 at low-elevation sites in the Southern Alps

We describe the recovery of three daily meteorological records for the southern Alps (Domodossola, Riva del Garda, and Rovereto), all starting in the second half of the nineteenth century. We use these new data, along with additional records, to study regional changes in the mean temperature and extreme indices of heat waves and cold spells frequency and duration over the period 1874–2015. The records are homogenized using subdaily cloud cover observations as a constraint for the statistical model, an approach that has never been applied before in the literature. A case study based on a record of parallel observations between a traditional meteorological window and a modern screen shows that the use of cloud cover can reduce the root-mean-square error of the homogenization by up to 30% in comparison to an unaided statistical correction. We find that mean temperature in the southern Alps has increased by 1.4°C per century over the analyzed period, with larger increases in daily minimum temperatures than maximum temperatures. The number of hot days in summer has more than tripled, and a similar increase is observed in duration of heat waves. Cold days in winter have dropped at a similar rate. These trends are mainly caused by climate change over the last few decades.

Functional sites involved in modulation of the GABAA receptor channel by the intravenous anesthetics propofol, etomidate and pentobarbital.

GABAA receptors are the major inhibitory neurotransmitter receptors in the brain and are the target for many clinically important drugs. Among the many modulatory compounds are also the intravenous anesthetics propofol and etomidate, and barbiturates. The mechanism of receptor modulation by these compounds is of mayor relevance. The site of action of these compounds has been located to subunit interfaces in the intra-membrane region of the receptor. In α1β2γ2 GABAA receptors there are five such interfaces, two β+/α- and one each of α+/β-, α+/γ- and γ+/β- subunit interfaces. We have used reporter mutations located in the second trans-membrane region in different subunits to probe the effects of changes at these subunit interfaces on modulation by propofol, etomidate and pentobarbital. We provide evidence for the fact that each of these compounds either modulates through a different set of subunit interfaces or through the same set of subunit interfaces to a different degree. As a GABAA receptor pentamer harbors two β+/α- subunit interfaces, we used concatenated receptors to dissect the contribution of individual interfaces and show that only one of these interfaces is important for receptor modulation by etomidate.

New Sites, New Challenges? Ethical and Methodological Questions in the Context of Research on Migration Exclusion

The challenges of research ethics and methodologies have been reflected on extensively, but – aside from the context of feminist methodologies – less so in relation to research on particular migration sites such as in transit, detention centres, at the borders or within migration administration. First attempts in this direction have been made (Düvell et al. 2010, Fresia et al. 2005, Riedner 2014, van Liempt/Bilger2009), however, more reflection and theorization is needed, considering the contested nature of these temporal and volatile sites. In this workshop, we thus aim at examining methodological as well as ethical questions that arise during field work: We attempt to reflect the power relations involved in the research process, the ethics of research design, the dissemination of research results, the question of gaining access to and – whenever necessary – staying in contact with our research subjects. How can we negotiate informed consent with subjects whose life is currently marked by transit and insecurity concerning their own future, and who are in an uncertain situation in which substantial information (legal, social, cultural etc.) is likely to be missing? How do we deal with the dilemma of possibly contributing to knowledge production that might facilitate removals and deportations in the future, considering that the reception of the results is not in the hands of the researchers? How do we deal with the anticipated as well as unexpected impacts of our research on social and political practice? Regarding fieldwork in state institutions, how do we negotiate the multiple loyalties we often find ourselves faced with as social researchers, both with the excluded migrants and with the authorities implementing the exclusions – two groupings considered to be opposite to each other (Lavanchy 2013)? Which different roles do researchers need to take on? The aim of our workshop is first and foremost to exchange experiences on fieldwork with others doing qualitative research on related topics and to consider its possible implications – including affective dimensions – for all participants involved in the research process: the migrants, the security staff of detention centres, its social workers, border police and bureaucrats and, last but not least, the researchers themselves. Furthermore, we generally wish to reflect upon the question of how best to conduct research in this contested field, applying an interdisciplinary perspective.

Macrofossils as records of plant responses to rapid Late Glacial climatic changes at three sites in the Swiss Alps

Plant macrofossils from the end of the Younger Dryas were analysed at three sites, Gerzensee (603 m asl), Leysin (1230 m asl), and Zeneggen (1510 m asl). For the first two sites an oxygen-isotope record is also available that was used to develop a time scale (Schwander et al., this volume); dates refer therefore to calibrated years according to the GRIP time scale. Around Gerzensee a pine forest with some tree birches grew during the Younger Dryas. With the onset of the isotopic shift initiating the rapid warming (about 11,535 cal. years before 1950), the pine forest became more productive and denser. At Leysin no trees except some juniper scrub grew during the Younger Dryas. Tree birches, pine, and poplar immigrated from lower altitudes and arrived after the end of the isotopic shift (about 11,487 B.P.), i.e., at the beginning of the Preboreal (at about 11,420 B.P.). Zeneggen is situated somewhat higher than Leysin, but single tree birches and pines survived the Younger Dryas at the site. At the beginning of the Preboreal their productivity and population densities increased. Simultaneously shifts from Nitella to Chara and from silt to gyttja are recorded, all indicating rapidly warming conditions and higher nutrient levels of the lake water (and probably of the soils in the catchment). At Gerzensee the beginning of the Younger Dryas was also analysed: the beginning of the isotopic shift correlates within one sample (about 15 years) to rapid decreases of macrofossils of pines and tree birches.

Pond-Breeding Amphibian Species Distributions in a Beaver-Modified Landscape, Acadia National Park, Mount Desert Island, Maine

In order to maintain pond-breeding amphibian species richness, it is important to understand how both natural and anthropogenic disturbances affect species assemblages and individual species distributions both at the scale of individual ponds and at a larger landscape scale. The goal of this project was to investigate what characteristics of ponds and the surrounding wetland landscape were most effective in predicting pond-breeding species richness and the individual occurrence of wood frog (Rana sylvatica), bullfrog (Rana catesbeiana) and pickerel frog (Rana palustris) breeding sites in a beaver-modified landscape and how this landscape has changed over time. The wetland landscape of Acadia National Park was historically modified by the natural disturbance cycles of beaver (Castor cazadensis), and since their reintroduction to the island in 1921, beaver have played a large role in creating and maintaining palustrine wetlands. In 2000 and 2001, I studied pond-breeding amphibian assemblages at 71 palustrine wetlands in Acadia National Park, Mount Desert Island, Maine. I determined breeding presence of 7 amphibian species and quantified 15 variables describing local pond conditions and characteristics of the wetland landscape. I developed a priori models to predict sites with high amphibian species and used model selection with Akaike's Information Criterion (AIC) to identify important variables. Single species models were also developed to predict wood frog, bullfrog and pickerel frogs breeding presence. The variables for wetland connectivity by stream corridors and the presence of beaver disturbance were the most effective variables to predict sites with high amphibian richness. Wood frog breeding was best predicted by local scale variables describing temporary, fishless wetlands and the absence of active beaver disturbance. Abandoned beaver sites provided wood frog breeding habitat (70%) in a similar proportion to that found in non beaver-influenced sites (79%). In contrast, bullfrog breeding presence was limited to active beaver wetlands with fish and permanent water, and 80% of breeding sites were large (≥2ha in size). Pickerel frog breeding site selection was predicted best by the connectivity of sites in the landscape by stream corridors. Models including the presence of beaver disturbance, greater wetland perimeter and greater depth were included in the confidence set of pickerel frog models but showed considerably less support. Analysis of historic aerial photographs showed an 89% increase in the total number of ponded wetlands available in the landscape between the years of 1944 and 1997. Beaver colonization generally converted forested wetlands and riparian areas to open water and emergent wetlands. Temporal colonization of beaver wetlands favored large sites low in the watersheds and sites that were impounded later were generally smaller, higher in the watershed, and more likely to be abandoned. These results suggest that beaver have not only increased the number of available breeding sites in the landscape for pond-breeding amphibians, but the resulting mosaic of active and abandoned beaver wetlands also provides suitable breeding habitat for species with differing habitat requirements.

Variables affecting the stability of multiple cloning sites and hairpin structures in retroviral vectors

Retroviruses are RNA viruses that replicate through a double-stranded DNA intermediate. The viral enzyme reverse transcriptase copies the retroviral genomic RNA into this DNA intermediate through the process of reverse transcription. Many variables can affect the fidelity of reverse transcriptase during reverse transcription, including specific sequences within the retroviral genome. ^ Previous studies have observed that multiple cloning sites (MCS) and sequences predicted to form stable hairpin structures are hotspots for deletion during retroviral replication. The studies described in this dissertation were performed to elucidate the variables that affect the stability of MCS and hairpin structures in retroviral vectors. Two series of retroviral vectors were constructed and characterized in these studies. ^ Spleen necrosis virus-based vectors were constructed containing separate MCS insertions of varying length, orientation, and symmetry. The only MCS that was a hotspot for deletion formed a stable hairpin structure. Upon more detailed study, the MCS previously reported as a hotspot for deletion was found to contain a tandem linker insertion that formed a hairpin structure. Murine leukemia virus-based vectors were constructed containing separate sequence insertions of either inverted repeat symmetry (122IR) that could form a hairpin structure, or little symmetry (122c) that would form a less stable structure. These insertions were made into either the neomycin resistance marker ( neo) or the hygromycin resistance marker (hyg) of the vector. 122c was stable in both neo and hyg, while 122IR was preferentially deleted in neo and was remarkably unstable in hyg. ^ These results suggest that MCS are hotspots for deletion in retroviral vectors if they can form hairpin structures, and that hairpin structures can be highly unstable at certain locations in retroviral vectors. This information may contribute to improved design of retroviral vectors for such uses as human gene therapy, and will contribute to a greater understanding of the basic science of retroviral reverse transcription. ^

Using GIS methods to assess the occurrence of neural tube defect births by residential proximity at conception to hazardous waste sites

In the United States, approximately 4,000 pregnancies each year are affected by the two most common birth defects, spina bifida and anencephaly. Studies have shown that exposure to environmental chemicals before and after conception may adversely affect reproduction by inducing cell death or dysfunction, which leads to infertility, fetal loss, lowered weight at birth, or birth anomalies in the offspring. The objective of the study was to evaluate the relationship between Neural Tube Defect births and residence at conception in proximity to hazardous waste sites in the Texas-Mexico border region between 1993 and 2000. ^ The study design was a nested matched case-control and utilized secondary data from a project, “The role of chemical and biological factors in the etiology of neural tube birth defects births along the Texas-Mexico Border” (Irina Cech, Principal Investigator). Geographic Information Systems (GIS) database methods were used to compare Neural Tube Defects cases to controls on status of conception residence occurring within a one-mile radius from hazardous waste sites, as compared to conception residence further away. Information on the exposures was obtained from the OnTarget Database and Environment Protection Agency website. Conditional logistic regression was used for the matched case-control study to investigate the relationship between an outcome of being a case or a control and proximity to hazardous waste sites. ^ The result of the study showed a 36 percent non-significant increased risk of having an NTD birth associated with maternal proximity to abandoned hazardous waste sites (95% CI = 0.62–3.02). In addition, there was a 24% non-significant elevated risk of having an NTD birth when living in proximity to air pollutant sites than when living further away (95% CI = 0.67–2.32). Although this study did not find statistically significant associations, it will expand on the existing knowledge of the relationship between NTD and proximity to hazardous waste sites. ^

Controlling Bacterial Wilt in Muskmelon with Perimeter Trap Cropping

Perimeter trap cropping (PTC) involves planting one or more rows of a cucurbit crop that is highly attractive to cucumber beetles around the border of a main cucurbit cash crop that is less attractive to the beetles. Cucumber beetles attempting to migrate into the field are concentrated in the relatively more attractive border crop, where they can be controlled by insecticides. In New England, perimeter trap cropping using Blue Hubbard squash as the border crop around pumpkin, cucumber, or butternut squash controlled cucumber beetle/bacterial wilt with as few as one border spray of insecticide. This strategy reduced insecticide use on the main crop by up to 94 percent, nearly eliminating sprays on the main cash crop. In on-farm trials, 8 of 10 Massachusetts growers found that using perimeter trap cropping saved them money. The same tactic also effectively managed cucumber beetles on muskmelon and squash in Oklahoma.