Flow cytometric analysis of cellular immune responses in dogs experimentally infected with a North American isolate of Leishmania infantum

Canine leishmaniasis caused by Leishmania infantum is endemic in the foxhound population in North America. Studies of canine leishmaniasis in the Mediterranean basin indicate a role for both CD4+ and CD8+ lymphocytes with clinical illness and in asymptomatic dogs. Limited information is available on the strain of L. infantum infecting foxhounds in North America. The present study investigated changes in cellular immune responses in dogs experimentally infected with 1x10(7) (low dose, LD; N=4) or 2x10(8) (high dose, HD; N=4) promastigotes of a United States isolate of L. infantum and control dogs (N=2) for 72 weeks. Density gradient separation was used to enrich for peripheral blood lymphocytes from canine blood. Lymphocyte subsets (CD4+ and CD8+) were quantified by flow cytometric analysis. Lymphocyte population expression levels over the course of the present study were compared to clinical status of the dog and antibody responses in infected and control dogs. No significant differences (P>0.05) were observed in either CD4+ or CD8+ lymphocyte expression in of the groups over the experimental period. This study suggests that the cellular immune responses to North American L. infantum in experimentally infected dogs may differ from other strains of L. infantum.     

Vitrification of Immature Feline Oocytes with a Commercial Kit for Bovine Embryo Vitrification

The aim of this study was to evaluate the suitability of a commercial kit for bovine embryo vitrification for cryopreserving cat oocytes and to evaluate comparatively the effects of its use with slow freezing procedure on cryotolerance in terms of morphology and oocyte resumption of meiosis. Germinal vesicle stage oocytes isolated from cat ovaries were either vitrified (n = 72) using a vitrification kit for bovine embryo or slow frozen (n = 69) by exposing oocyte to ethylene glycol solution before being transferred to a programmable embryo freezer. After thawing and warming, oocytes were cultured for 48 h and then were examined for meiosis resumption using bisbenzimide fluorescent staining (Hoechst 33342). Fresh immature oocytes (n = 92) were used as the control group. The proportion of oocytes recovered in a morphologically normal state after thawing/warming was significantly higher in frozen oocytes (94.5%) than in the vitrified ones (75%, p < 0.01). Morphological integrity after culture was similar in vitrified (73.6%) and slow frozen oocytes (76.8%); however, only 37.5% of the morphologically normal oocytes resumed meiosis after vitrification compared to 60.9% of those submitted to slow freezing procedure (p < 0.01). Fresh oocytes showed higher morphological integrity (91.3%) and meiosis resumption rates (82.6%, p < 0.002) than cryopreserved oocytes, irrespective of the procedure used. These results suggest that immature cat oocytes vitrified with a kit for bovine embryos retain their capacity to resume meiosis after warming and culture, albeit at lower rates than slow frozen oocytes. Vitrification and slow freezing methods show similar proportions of oocytes with normal morphology after culture, which demonstrate that thawed and warmed oocytes that resist to cryodamage have the same chances to maintain their integrity after 48 h of culture.     

Impact of sustained deficit irrigation on spearmint (<Emphasis Type="Italic">Mentha spicata</Emphasis> L.) biomass production,oil yield,and oil quality

Crop response to deficit irrigation is an important consideration for establishing irrigation management strategies when water supplies are limited. This study evaluated the response of native spearmint to water deficits applied using overhead sprinklers in eastern Washington, US. Five levels of irrigation were applied ranging from full irrigation (100%) to 5% of weekly averaged full crop water needs. Soil water monitoring with soil water balance was used to estimate soil water deficits for irrigation scheduling and soil water use. Mint oil yields, oil components, dry matter production, and the water-use efficiency of the spearmint were assessed. There was significant reduction in fresh mint hay (harvested biomass) yield with increasing water deficit. However, spearmint oil yields remained generally uniform across irrigation treatments at the first cutting but decreased at the driest plots during the second harvest due to a loss of plant stand. The wet harvest index and water-use efficiency improved significantly for both harvests with increasing water deficit. Hay yield, oil yield, wet harvest index, and water-use efficiency are pooled across sides and replicate blocks to provide trends with changes in maximum evapotranspiration. The three major monoterpenes show changes suggesting less mature oil yields. The study demonstrates the feasibility of sustaining native spearmint yields under managed deficit irrigations for deficits not lower than 0.5 ETc.     

High-quality binary protein interaction map of the yeast interactome network

Current yeast interactome network maps contain several hundred molecular complexes with limited and somewhat controversial representation of direct binary interactions. We carried out a comparative quality assessment of current yeast interactome data sets, demonstrating that high-throughput yeast two-hybrid (Y2H) screening provides high-quality binary interaction information. Because a large fraction of the yeast binary interactome remains to be mapped, we developed an empirically controlled mapping framework to produce a "second-generation" high-quality, high-throughput Y2H data set covering approximately 20% of all yeast binary interactions. Both Y2H and affinity purification followed by mass spectrometry (AP/MS) data are of equally high quality but of a fundamentally different and complementary nature, resulting in networks with different topological and biological properties. Compared to co-complex interactome models, this binary map is enriched for transient signaling interactions and intercomplex connections with a highly significant clustering between essential proteins. Rather than correlating with essentiality, protein connectivity correlates with genetic pleiotropy.     

Elemental concentrations in fresh snowfall across a regional transect in the northeastern U.S.: Apparent sources and contribution to acidity

Fresh snowfall was collected on the surface of 8 lakes across a 350 km west-east transect from northeastern New York state to the coast of Maine after a single storm. In addition, every snowfall event during the winter of 1993 was collected on a single lake near the center of the transect. Across the transect, midwestern sources appear to dominate Pb and Cd concentrations, while Sb appears to be derived from midwestern sources as well as local and/or industrial East Coast sources. In all samples, the highest Na, Cl and Mg concentrations reflect a marine influence, but at some transect sites roadspray aerosol appears to contribute to Na and Cl concentrations. The regional pattern of Ca, K, Mn and Sr concentrations and Mn/Sr ratios indicate that woodsmoke may be an important winter source of these elements at some sites. In all samples, H⁺ is strongly correlated with NO₃ ^? (R² = 0.97) and mean NO₃ ^?/SO₄ ^2? molar ratios of 6.4 for transect samples, and 4.7 for temporal samples, are higher than mean NO₃ ^?/SO₄ ^2? reported for other precipitation studies in the same region. The contribution of NO₃ ^? to the snowpack greatly exceeds that of SO₄ ^2?, and may be a major source of acidity in aquatic ecosystems during snowmelt.     

Retrograde signaling by Syt 4 induces presynaptic release and synapse-specific growth

The molecular pathways involved in retrograde signal transduction at synapses and the function of retrograde communication are poorly understood. Here, we demonstrate that postsynaptic calcium 2+ ion (Ca2+) influx through glutamate receptors and subsequent postsynaptic vesicle fusion trigger a robust induction of presynaptic miniature release after high-frequency stimulation at Drosophila neuromuscular junctions. An isoform of the synaptotagmin family, synaptotagmin 4 (Syt 4), serves as a postsynaptic Ca2+ sensor to release retrograde signals that stimulate enhanced presynaptic function through activation of the cyclic adenosine monophosphate (cAMP)-cAMP-dependent protein kinase pathway. Postsynaptic Ca2+ influx also stimulates local synaptic differentiation and growth through Syt 4-mediated retrograde signals in a synapse-specific manner.     

Thermal Characterization of Alkali Treated Kenaf Fibers and Kenaf-Epoxy Composites

Chemical treatment of natural fibers is a well-defined means of mechanical property improvement in natural fiberreinforced composites. An understanding of mechanical and thermal properties in these media is essential for evaluating heat transfer, thermal degradation, and overall performance of these composites over their product lifetime. However, very little information is available illustrating the effect of such treatment on the thermal properties of kenaf composites. Also, no study to date has reported the thermal conductivity of individual kenaf fibers. This study reports the effects of fiber treatment (in 6 % NaOH) on thermal transport in unidirectionally oriented kenaf-epoxy composites and individual kenaf fibers. The effective thermal conductivities and thermal diffusivities of chemically treated fiber composites show a general increase over untreated fiber composites (0.210 to 0.232 W/m/K at 28 °C, 0.206 to 0.234 W/m/K at 200 °C). This improvement may be attributed to improved interfacial contact between the fibers and epoxy matrix shown in microstructural images after chemical treatment. The thermal conductivity of individual fibers was evaluated at room temperature using two techniques. Results from both techniques showed slight increases after chemical treatment (0.58±0.53 to 1.0±0.13 W/m/K and 1.2±0.54 to 1.6±0.28 W/m/K) but lacked statistical significance. Any improvement in surface crystallinity after chemical treatment does not appear to affect overall fiber thermal conductivity. A better understanding of thermal transport in kenaf fibers and composites enables better estimation of the performance of these composites in different applications. Moreover, the thermal conductivities of individual fibers are useful in understanding the fiber’s contribution to conduction in different fiber reinforcement configurations.     

The use of high-energy electrons to depilate the breech of sheep

The use of high-energy electrons for permanently depilating areas of sheep skin was evaluated. The most effective dose of electrons for depilation was 17.5Gy. Histological changes in skin treated at this dose were examined in one sheep over 18 months and in 5 sheep over 89 d. Effects of treatment on bodyweight gains and fleece growth were examined by comparing the productivity of a further 5 sheep treated on the breech with high-energy electrons, with that of conventionally mulesed sheep (n = 5) and untreated controls (n = 5). Electron treatment resulted in immediate death of cells in the germinative region of the wool follicle bulbs. Within 10 d of treatment the treated areas were completely depilated. Wool follicle shafts in the treated areas regressed rapidly towards the epidermis and remained quiescent for the whole trial (89 d). A sheep treated 18 months previously has remained largely depilated, although a few sparse fibres are present. Epidermal acanthosis and orthokeratosis were present at 26 d after treatment. The thickened stratum corneum then sloughed off, but the epidermis remained acanthotic for the entire trial. Sweat glands and most sebaceous glands were destroyed by the treatment and were replaced by fibrotic, avascular tissue in the dermis. In all other respects the external appearance of the electron-treated breech was similar to that of mulesed sheep. There were no apparent side-effects of treatment. Neither mulesing nor electron treatment altered weight gains or fleece growth rates.     

Outplanting Wyoming Big Sagebrush Following Wildfire: Stock Performance and Economics

Finding ecologically and economically effective ways to establish matrix species is often critical for restoration success. Wyoming big sagebrush (Artemisia tridentata subsp. wyomingensis) historically dominated large areas of western North America, but has been extirpated from many areas by large wildfires; its re-establishment in these areas often requires active management. We evaluated the performance (survival, health) and economic costs of container and bare-root stock based on operational plantings of more than 1.5 million seedlings across 2 200 ha, and compared our plantings with 30 other plantings in which sagebrush survival was tracked for up to 5 yr. Plantings occurred between 2001 and 2007, and included 12 combinations of stock type, planting amendment, and planting year. We monitored 10 500 plants for up to 8 yr after planting. Survival to Year 3 averaged 21% and was higher for container stock (30%) than bare-root stock (17%). Survival did not differ among container-stock plantings, whereas survival of bare-root stock was sometimes enhanced by a hydrogel dip before planting, but not by mycorrhizal amendments. Most mortality occurred during the first year after planting; this period is the greatest barrier to establishment of sagebrush stock. The proportion of healthy stock in Year 1 was positively related to subsequent survival to Year 3. Costs were minimized, and survival maximized, by planting container stock or bare-root stock with a hydrogel dip. Our results indicate that outplanting is an ecologically and economically effective way of establishing Wyoming big sagebrush. However, statistical analyses were limited by the fact that data about initial variables (stock quality, site conditions, weather) were often unrecorded and by the lack of a replicated experimental design. Sharing consistent data and using an experimental approach would help land managers and restoration practitioners maximize the success of outplanting efforts.