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At least 397 records · Page 22Linked to original sources

Spatial genetic structure in two congeneric epiphytes with different dispersal strategies analysed by three different methods.

Three different approaches were used to assess the kinship structure of two epiphytic bryophytes, Orthotrichum speciosum and O. obtusifolium, that have different dispersal strategies. The two species were sampled in a 200 ha landscape where species occurrence and host trees had been mapped previously. Local environmental conditions at sampled trees were recorded and kinship between individuals was calculated based on amplified fragment length polymorphism (AFLP)-marker data. We did not detect any association between AFLP-markers and investigated environmental conditions. In both species, significant kinship coefficients were found between individuals up to 300-350 m apart which shows that both species have a restricted dispersal range. The spatial kinship structure was detected with both autocorrelation analysis and generalized additive models (GAMs), but linear regression failed to detect any structure in O. speciosum. Although the dioecious O. obtusifolium is currently the more common species it may, none the less, due to its restricted dispersal range and reproduction mode, become threatened in the future by current silvicultural practices which enhance the distance between host trees and decrease their life span. Finally, GAMs seem most appropriate for analysing spatial genetic structure because the effects of local environmental conditions and spatial structure can be analysed simultaneously, no assumption of a parametric form between kinship coefficient and distance is required, and spatial data resolution is not lost in the arbitrary choice of distance classes characterizing autocorrelation analysis.

Bryopsida↗

Using intrinsically fluorescent proteins for plant cell imaging.

The intrinsically fluorescent proteins (IFPs), such as the green, cyan and yellow fluorescent proteins, have revolutionized how we can image the dynamics of cellular events. Intrinsically fluorescent proteins have been used as reporter genes to monitor transcriptional regulation, as targeted markers for organelles and subcellular structures, in fusion proteins to directly observe protein motility and dynamics, and in sensors designed to show changes in cellular environments ranging from pH to protein kinase activity. The IFPs hold tremendous potential to reveal the dynamic processes that underlie plant cell function; however, as with all technology there are artifacts and pitfalls inherent in their use. In this review, we highlight some of the practical issues in using IFPs for live cell imaging. These include choice of the appropriate IFP, dealing with autofluorescence, photobleaching and phototoxicity, and application of approaches such as fluorescence resonance energy transfer (FRET), fluorescence lifetime imaging (FLIM) and fluorescence recovery after photobleaching (FRAP) to gain high-resolution data about protein dynamics within the cell. We also discuss some of the more common artifacts associated with these fluorescence imaging approaches and suggest controls that should help both spot these problems and suggest their solutions.

Fluorescence Resonance Energy Transfer↗

High-resolution typing of HLA-DRB1 locus in the Macedonian population.

The Macedonian population is of special interest for HLA anthropological study in the light of unanswered questions regarding its origin and relationship with other populations, especially the neighbouring Balkanians. Two studies have been performed to examine HLA molecular polymorphism in the Macedonian population, so far. The present study is the first to be performed in Macedonia using high-resolution sequence-based method for direct HLA typing. The study included 158 unrelated healthy volunteers of Macedonian origin and nationality, having a Christian Orthodox religion. After the simultaneous amplification of exon-2 on both HLA-DRB1 alleles, DNA sequencing was used for genotype assignment. In the 158 samples analysed, all 316 alleles were typed and a total of 29 different DRB1 alleles were detected, with DRB1*1601 being the most frequent allele (14.9%), followed by DRB1*1104 (13.9%). A phylogenetic tree constructed on the basis of the high-resolution data deriving from other populations revealed the clustering of Macedonians together with other Balkan populations (Greeks, Croats, Turks and Romanians) and Sardinians, close to another "European" cluster consisting of the Italian, French, Danish, Polish and Spanish populations. The included African populations grouped on the opposite side of the tree.

Alleles↗

How we do it: coblation tonsillectomy complication rates from a single ENT department compared with the National Prospective Tonsillectomy Audit.

Coblation tonsillectomy is a relatively new technique, the results of which need auditing within practising units, to justify its continued usage. * The National Prospective Tonsillectomy Audit provides an excellent source of data for individual units to compare their results to. * This retrospective audit of 391 coblation tonsillectomies shows that our units haemorrhage and return to theatre rates are similar to the National rates for cold steel & ties data. * Resolution of training issues and patient selection may lead to increased use of this technique.

Adolescent↗

Conservation conflicts across Africa.

There is increasing evidence that areas of outstanding conservation importance may coincide with dense human settlement or impact. We tested the generality of these findings using 1 degree-resolution data for sub-Saharan Africa. We find that human population density is positively correlated with species richness of birds, mammals, snakes, and amphibians. This association holds for widespread, narrowly endemic, and threatened species and looks set to persist in the face of foreseeable population growth. Our results contradict earlier expectations of low conflict based on the idea that species richness decreases and human impact increases with primary productivity. We find that across Africa, both variables instead exhibit unimodal relationships with productivity. Modifying priority-setting to take account of human density shows that, at this scale, conflicts between conservation and development are not easily avoided, because many densely inhabited grid cells contain species found nowhere else.

Africa South of the Sahara↗

Systematic variation in DNA length yields highly ordered repressor-operator cocrystals.

Crystals have been grown that contain the operator-binding domain of the lambda repressor and the lambda operator site OL1. Crystallization conditions were tested with a set of DNA fragments, ranging in length from 17 to 23 base pairs. The best crystals were grown with a 20-base pair DNA fragment. These crystals have space-group symmetry P2I, with unit cell dimensions a = 37.1 A, b = 68.8 A, c = 56.8 A, and a beta angle of 91.5 degrees. They diffracted to at least 2.5 A resolution. High resolution data from these crystals should allow the direct determination of how a repressor recognizes its operator site.

Base Sequence↗

Structural and thermodynamic basis for the binding of TMC114, a next-generation human immunodeficiency virus type 1 protease inhibitor.

TMC114, a newly designed human immunodeficiency virus type 1 (HIV-1) protease inhibitor, is extremely potent against both wild-type (wt) and multidrug-resistant (MDR) viruses in vitro as well as in vivo. Although chemically similar to amprenavir (APV), the potency of TMC114 is substantially greater. To examine the basis for this potency, we solved crystal structures of TMC114 complexed with wt HIV-1 protease and TMC114 and APV complexed with an MDR (L63P, V82T, and I84V) protease variant. In addition, we determined the corresponding binding thermodynamics by isothermal titration calorimetry. TMC114 binds approximately 2 orders of magnitude more tightly to the wt enzyme (K(d) = 4.5 x 10(-12) M) than APV (K(d) = 3.9 x 10(-10) M). Our X-ray data (resolution ranging from 2.2 to 1.2 A) reveal strong interactions between the bis-tetrahydrofuranyl urethane moiety of TMC114 and main-chain atoms of D29 and D30. These interactions appear largely responsible for TMC114's very favorable binding enthalpy to the wt protease (-12.1 kcal/mol). However, TMC114 binding to the MDR HIV-1 protease is reduced by a factor of 13.3, whereas the APV binding constant is reduced only by a factor of 5.1. However, even with the reduction in binding affinity to the MDR HIV protease, TMC114 still binds with an affinity that is more than 1.5 orders of magnitude tighter than the first-generation inhibitors. Both APV and TMC114 fit predominantly within the substrate envelope, a property that may be associated with decreased susceptibility to drug-resistant mutations relative to that of first-generation inhibitors. Overall, TMC114's potency against MDR viruses is likely a combination of its extremely high affinity and close fit within the substrate envelope.

Binding Sites↗

Structural relationships in the two-zinc insulin hexamer.

The refinement of the crystal structure of two-Zn pig insulin using 1.5-A (1A = 0.1 nm) resolution data by Fourier and fast Fourier least-squares methods allows us to make detailed comparisons between the two independent molecules present in the two-Zn insulin dimer and to describe their interactions in the monomer, dimer, and hexamer. The main chain structures for the two molecules agree well except at the N terminus of the A chain and the C terminus of the B chain. The residues along the line of the local two-fold axes, apart from the B25 side chain, conform extremely closely to the two-fold symmetry, although the discrepancies are much more apparent away from this axis. The ability of the insulin molecule to adopt different conformations may be an important factor in the expression of its biological activity.

Animals↗

Methodology of in vivo human sodium MR imaging at 1.5 T.

The methodology of sodium-23 (Na-23) imaging is reported in relationship to the physiological factors that determine the chemical environment of the Na-23 nucleus. Contrast resolution is given as a function of imaging time and spatial resolution. Data showing the optimal relaxation time for sodium imaging are given, and the linear quantitative relationship between sodium concentration and voxel intensity for our imaging system is confirmed. The major problem facing in vivo sodium imaging is the ability to differentiate intracellular sodium from extracellular sodium. The sodium in blood serum (extracellular) and packed red blood cells (intracellular) both exhibit biexponential T2 decay. These results indicate that T2 measurements alone will be insufficient for discriminating extracellular from intracellular sodium. Instead, other methods based on the underlying physiological properties of in vivo sodium imaging, such as the diffusion coefficient, will be necessary to truly separate extracellular from intracellular sodium.

Brain Neoplasms↗

Vascular emergencies of the thorax after blunt and iatrogenic trauma: multi-detector row CT and three-dimensional imaging.

Multi-detector row computed tomographic (CT) angiography is an effective modality for vascular imaging in the thorax. It allows acquisition of high-resolution data sets during a single breath hold, making it the preferred method for evaluation of patients with acute vascular disease. In contrast to conventional angiography, multirow CT angiography not only depicts the vessels but also allows assessment of adjacent structures. Multirow CT angiography with two- and three-dimensional reformation can be used to diagnose vascular emergencies of the thorax after blunt and iatrogenic trauma. These include incomplete and complete aortic rupture; traumatic aortic dissection; arterial dissection and rupture after minor trauma in patients with Ehlers-Danlos syndrome; traumatic intramural hematoma; pseudoaneurysm after endovascular repair; injuries due to Swan-Ganz catheters; complications of central venous cannulation, pacemaker implantation, and percutaneous pericardial drainage; and foreign-body embolism. The diagnoses can be established with multirow CT angiography in the emergency department. Thus, the time to diagnosis can be considerably decreased by obviating conventional angiography. Knowledge of the CT findings in various vascular conditions is essential to make use of multirow CT angiography in combination with two- and three-dimensional reformation as an efficient and accurate diagnostic tool in emergency radiology.

Adult↗

Multi-detector row CT and postprocessing techniques in the assessment of diffuse lung disease.

Many acute and chronic lung diseases are characterized by diffuse infiltration of the lung parenchyma. High-resolution computed tomography (CT) has been widely accepted as the imaging standard of reference for the assessment of these diseases. However, only approximately 10% of the lung parenchyma is scanned with high-resolution CT, and characteristic foci of disease may be missed. With use of the established characteristic high-resolution CT patterns, multi-detector row chest CT has revolutionized the evaluation of diffuse lung disease. Multi-detector row CT generates isotropic volumetric high-resolution data, allowing contiguous three-dimensional (3D) visualization of the lung parenchyma, with the capacity to create high-quality two-dimensional (2D) and 3D reformatted images. Minimum intensity projection is the postprocessing technique of choice for the detection and characterization of most patterns of diffuse lung disease. Maximum intensity projection (MIP) allows the detection and characterization of micronodules; the recognition of enlarged pulmonary veins, which is extremely useful in the diagnosis of pulmonary edema and the assessment of mosaic perfusion; and differentiation between perilymphatic, miliary, and centrilobular distribution. MIP can also help differentiate between constrictive bronchiolitis and mixed emphysema. Two-dimensional reformatted images are now of equal importance with the 2D axial images in diagnosing specific diffuse lung diseases. In the future, 3D reformatted images may be used to help quantify these disorders.

Adult↗

Ancient DNA and Human Physiology.

Ancient DNA (aDNA) enables the reconstruction of chronologically sampled genomes from ancient humans, animals, plants, pathogens, and microorganisms, as well as environmental DNA, providing a record of biological changes through time. Improvements in short and degraded DNA extraction methods and low-cost sequencing now enable the generation of broad, cross-regional datasets that expand evolutionary analyses from past population demography to biological mechanisms. By tracking temporal shifts of allele frequencies, integrating functional genomics resources (e.g., gene expression, chromatin structure variation), modeling population demography to separate selection from genetic drift, and aligning genetic changes with archaeological, cultural, and climatic data, aDNA has the potential to link sequence variation to physiological function within their temporal and environmental contexts. In this review, we summarize illustrative case studies from aDNA research spanning complex traits, dietary adaptations, and responses to pathogens and other environmental changes, showing how human biology has evolved under multiple selective pressures through time. These dated signals help triage experimental work and expose mechanisms that are rare or absent in living cohorts. Although some challenges remain, such as geographic and temporal sampling disparities, limitations in data resolution and variant detection, and genotype-phenotype uncertainties, rapid methodological progress and stronger ethical frameworks are expanding what can be inferred, making aDNA a promising tool for refining physiological pathways, their timing, and their drivers.

Humans↗

Microfluorimetric scanning of sympathetic nerve fibers. An improved method to quantitate formaldehyde induced fluorescence of biogenic amines.

A two dimensional scanning procedure was developed for studying quantitative aspects of formaldehyde induced fluorescence (FIF) from noradrenaline containing nerve fibers. A computer (PDP 12) controlled scanning device was equipped with a cooled photomultiplier, a 0.5 micron scanning stage and a 0.5 micron measuring spot. Photodecomposition was reduced by a high scanspeed (100/sec) and a small excitation field. Digitized images were obtained with a high resolution. Data (10,000/scan) of large scan areas (2500 micron2) in the iris of the rat were transformed into fluorescence histograms, which allowed statistical evaluation. Visible changes induced in the histochemical preparation by administration of drugs (reserpine, L-Dopa) were reflected in characteristic changes in the histograms. A minor reduction in noradrenaline content, which was too small to be detected by visual observation of the microscopical image and by classical microfluorimetric methods, did evoke a significant change in the histograms. It is concluded that microfluorimetric scanning is a very sensitive technique for detecting changes of formaldehyde induced fluorescence from neuronal networks.

Adrenergic Fibers↗

Solar ultraviolet-B exposure and cancer incidence and mortality in the United States, 1993-2002.

BACKGROUND: An inverse relationship between solar ultraviolet-B (UV-B) exposure and non-skin cancer mortality has long been reported. Vitamin D, acquired primarily through exposure to the sun via the skin, is believed to inhibit tumor development and growth and reduce mortality for certain cancers. METHODS: We extend the analysis of this relationship to include cancer incidence as well as mortality, using higher quality and higher resolution data sets than have typically been available. Over three million incident cancer cases between 1998 and 2002 and three million cancer deaths between 1993 and 2002 in the continental United States were regressed against daily satellite-measured solar UV-B levels, adjusting for numerous confounders. Relative risks of reduced solar UV-B exposure were calculated for thirty-two different cancer sites. RESULTS: For non-Hispanic whites, an inverse relationship between solar UV-B exposure and cancer incidence and mortality was observed for ten sites: bladder, colon, Hodgkin lymphoma, myeloma, other biliary, prostate, rectum, stomach, uterus, and vulva. Weaker evidence of an inverse relationship was observed for six sites: breast, kidney, leukemia, non-Hodgkin lymphoma, pancreas, and small intestine. For three sites, inverse relationships were seen that varied markedly by sex: esophagus (stronger in males than females), gallbladder (stronger in females than males), and thyroid (only seen in females). No association was found for bone and joint, brain, larynx, liver, nasal cavity, ovary, soft tissue, male thyroid, and miscellaneous cancers. A positive association between solar UV-B exposure and cancer mortality and incidence was found for anus, cervix, oral cavity, melanoma, and other non-epithelial skin cancer. CONCLUSION: This paper adds to the mounting evidence for the influential role of solar UV-B exposure on cancer, particularly for some of the less-well studied digestive cancers. The relative risks for cancer incidence are similar to those for cancer mortality for most sites. For several sites (breast, colon, rectum, esophagus, other biliary, vulva), the relative risks of mortality are higher, possibly suggesting that the maintenance of adequate vitamin D levels is more critical for limiting tumor progression than for preventing tumor onset. Our findings are generally consistent with the published literature, and include three cancer sites not previously linked with solar UV-B exposure, to our knowledge: leukemia, small intestine, and vulva.

Epidemiologic Studies↗

Synaptogenesis in the giant-fibre system of Drosophila: interaction of the giant fibre and its major motorneuronal target.

The tergotrochanteral (jump) motorneuron is a major synaptic target of the Giant Fibre in Drosophila. These two neurons are major components of the fly's Giant-Fibre escape system. Our previous work has described the development of the Giant Fibre in early metamorphosis and the involvement of the shaking-B locus in the formation of its electrical synapses. In the present study, we have investigated the development of the tergotrochanteral motorneuron and its electrical synapses by transforming Drosophila with a Gal4 fusion construct containing sequences largely upstream of, but including, the shaking-B(lethal) promoter. This construct drives reporter gene expression in the tergotrochanteral motorneuron and some other neurons. Expression of green fluorescent protein in the motorneuron allows visualization of its cell body and its subsequent intracellular staining with Lucifer Yellow. These preparations provide high-resolution data on motorneuron morphogenesis during the first half of pupal development. Dye-coupling reveals onset of gap-junction formation between the tergotrochanteral motorneuron and other neurons of the Giant-Fibre System. The medial dendrite of the tergotrochanteral motorneuron becomes dye-coupled to the peripheral synapsing interneurons between 28 and 32 hours after puparium formation. Dye-coupling between tergotrochanteral motorneuron and Giant Fibre is first seen at 42 hours after puparium formation. All dye coupling is abolished in a shaking-B(neural) mutant. To investigate any interactions between the Giant Fibre and the tergotroachanteral motorneuron, we arrested the growth of the motorneuron's medial neurite by targeted expression of a constitutively active form of Dcdc42. This results in the Giant Fibre remaining stranded at the midline, unable to make its characteristic bend. We conclude that Giant Fibre morphogenesis normally relies on fasciculation with its major motorneuronal target.

Animals↗

Polycomb group gene silencing proteins are concentrated in the perichromatin compartment of the mammalian nucleus.

Human Polycomb group (PcG) proteins are involved in cell-type-dependent epigenetic gene silencing in an evolutionarily conserved manner. We have analysed the subnuclear localisation of these regulatory proteins in two different human cell lines and in rat liver tissue by means of light and electron immunomicroscopy using specific antibodies. We find that the PcG proteins HPC2, HPH1, BMI1 and RING1 are highly concentrated in the perichromatin compartment, situated at the surface of condensed chromatin domains. This compartment was demonstrated earlier to be the nuclear site where most pre-mRNA synthesis takes place. Interestingly, these PcG proteins are virtually absent from the interior of condensed chromatin areas. The present observations therefore show that transcriptionally active and PcG-silenced loci occur within the same spatially limited nuclear domain. Our novel high-resolution data strongly support the idea that epigenetic PcG-mediated gene silencing is a local event, rather than affecting large chromatin domains. In addition to being associated with the perichromatin region, PcG proteins also occur in the interchromatin space. Implications of these observations for higher order chromatin structure and for the mechanisms of PcG-mediated gene silencing are discussed.

Animals↗

A continental risk map for malaria mosquito (Diptera: Culicidae) vectors in Europe.

Although malaria was officially declared eradicated from Europe in 1975, its former vectors, mainly members of the Anopheles maculipennis (Meigen) complex, are still distributed throughout the continent. The present situation of Anophelism without malaria indicates that current socio-economic and environmental conditions maintain the basic case reproduction number, Ro, below 1. Recently, it has been speculated that predicted climate changes may increase anopheline abundance and biting rates (as well as reduce the Plasmodium parasite extrinsic incubation period), allowing the reemergence of malaria transmission in Europe. As a preliminary step toward predicting future scenarios, we have constructed models to test whether the current distribution of the five former European malaria vectors [An. atroparvus (Van Thiel),An. labranchiae (Falleroni), An. messeae (Swellengrebel & De Buck), An. sacharovi (Favr) and An. superpictus (Grassi)] can be explained by environmental parameters, including climate. Multivariate logistic regression models using climate surfaces derived from interpolation of meteorological station data (resolution 0.5 x 0.5 degrees) and remotely sensed land cover (resolution 1 x 1 km) were fitted to 1,833 reported observations of the presence and absence of each species across Europe. These relatively crude statistical models predicted presence and absence with a sensitivity of 74-85.7% and specificity of 73.4-98.1% (with climate a significantly better predictor than land cover type). A geographically independent validation of the models gave a sensitivity of 72.9-88.5% and a specificity of 72.7-99.6%. This allowed us to generate risk maps for each species across Europe. Assuming that high risk equates with the potential for high abundance, these models should permit the development of risk maps for European mosquitoes under future climate scenarios. These techniques would be equally useful for estimating the risk of reemergence in other nonendemic areas such as the United States and Australia, as well as changes to risk within endemic areas.

Animals↗

Spatial analysis of land use impact on ground water nitrate concentrations.

In spatial analyses of causes or health effects of environmental pollutants, small units of analyses are usually preferred for internal environmental homogeneity reasons but can only be done when fine resolution data are available for most units. Objectives of this study were to determine which land use practices were spatially associated with ground water nitrate concentrations across Prince Edward Island (PEI), Canada, and which spatial aggregation is the preferred unit of analyses. Nitrate concentrations were determined for 4855 samples from private wells. Validated field-by-field land use data were available. Average nitrate concentration and percentage of area for the 14 major land use categories in PEI were determined for each of three spatial aggregations: watersheds based on topography and hydrology; freeform polygon boundaries based on similar neighboring nitrate concentrations; and 500-m buffer zones around each well. Results showed that the percentages of potato, grain, and hay coverage were positive predictors of ground water nitrate concentrations. Percentage of blueberry was a marginally significant negative predictor in the watershed and freeform polygon models, and percentage of residential coverage was a positive predictor in the freeform polygon and buffer zone models. Spatial autocorrelation was present in the freeform polygon and buffer zone models even after land use was taken into account. In conclusion, analyses based on watersheds produced the best predictive model with the percentages of land cover of potato, hay, and grain being significantly associated with ground water nitrate concentrations, and the percentages of blueberry, clear-cut woodland, and other agriculture being marginally significant.

Agriculture↗