Comparative nitric oxide production by LPS-stimulated monocyte-derived macrophages from Ovis canadensis and Ovis aries

Bighorn sheep are more susceptible to respiratory infection by Mannheimia haemolytica than are domestic sheep. In response to bacterial challenge, macrophages produce a number of molecules that play key roles in the inflammatory response, including highly reactive nitrogen intermediates such as nitric oxide (NO). Supernatants from monocyte-derived macrophages cultured with M. haemolytica LPS were assayed for nitric oxide activity via measurement of the NO metabolite, nitrite. In response to LPS stimulation, bighorn sheep macrophages secreted significantly higher levels of NO compared to levels for non-stimulated macrophages. In contrast, levels of NO produced by domestic sheep macrophages in response to M. haemolytica LPS did not differ from levels detected in non-stimulated cell cultures. Nitrite levels detected in supernatants of LPS-stimulated bighorn macrophage cultures treated with an inducible nitric oxide synthase (INOS) inhibitor, N(G)-monomethyl-L-arginine, were similar to that observed in non-stimulated cultures indicating a role for the iNOS pathway.     

Evaluation of a vessel-sealing device for laparoscopic granulosa cell tumor removal in standing mares

OBJECTIVE: To describe a laparoscopic technique for granulosa cell tumor removal using a vessel sealing device (LigaSuretrade mark) in standing mares. STUDY DESIGN: Retrospective study. ANIMALS: Eight mares (8-24 years old; weighing, 406-525 kg). METHODS: Before surgery, ovarian size and adjacent body wall thickness was determined by ultrasonography. Mares were sedated and after local anesthesia (inverted L and local infiltration), laparoscopic cannulation was performed without insufflation. The mesovarium was anesthetized and the LigaSure instrument applied to the mesovarium for hemostasis and resection to remove the affected ovary. Mares were hospitalized for 24 hours before discharge. RESULTS: Median ovarian diameter was 10.5 cm (range, 6-14 cm). Median surgery time was 75 minutes (range, 40-180 minutes). Hemostasis was achieved using the LigaSure device in all mares. Median length of the abdominal wall incision made to remove the ovary was 13 cm (range, 5-17 cm); no incisional complications occurred. CONCLUSIONS: The LigaSure vessel sealing device provided adequate hemostasis for removal of larger neoplastic ovaries in standing mares. CLINICAL RELEVANCE: Concerns of ligature placement can be alleviated by use of the LigaSure device and standing laparoscopic technique provides excellent observation of the surgical field ensuring hemostasis.     

Black cohosh (Actaea racemosa, Cimicifuga racemosa) behaves as a mixed competitive ligand and partial agonist at the human mu opiate receptor

Black cohosh is a commonly used botanical dietary supplement for the treatment of climacteric complaints. Because the opiate system in the brain is intimately associated with mood, temperature, and sex hormonal levels, the activity of black cohosh extracts at the human mu opiate receptor (hMOR) expressed in Chinese hamster ovary cells was investigated. The 100% methanol, 75% ethanol, and 40% 2-propanol extracts of black cohosh effectively displaced the specific binding of [3H]DAMGO to hMOR. Further studies of the clinically used ethanol extract indicated that black cohosh acted as a mixed competitive ligand, displacing 77 +/- 4% [3H]DAMGO to hMOR (Ki = 62.9 microg/mL). Using the [35S]GTPgammaS assay, the action of black cohosh was found to be consistent with an agonist, with an EC50 of 68.8 +/- 7.7 microg/mL. These results demonstrate for the first time that black cohosh contains active principle(s) that activate hMOR, supporting its beneficial role in alleviating menopausal symptoms.     

Common Kibra alleles are associated with human memory performance

Human memory is a polygenic trait. We performed a genome-wide screen to identify memory-related gene variants. A genomic locus encoding the brain protein KIBRA was significantly associated with memory performance in three independent, cognitively normal cohorts from Switzerland and the United States. Gene expression studies showed that KIBRA was expressed in memory-related brain structures. Functional magnetic resonance imaging detected KIBRA allele-dependent differences in hippocampal activations during memory retrieval. Evidence from these experiments suggests a role for KIBRA in human memory.     

Growth and phenology variation in progeny of Scots pine seed orchards and commercial seed stands

Tree improvement in Poland has been most advanced for Scots pine, but existing seed orchards have not been progeny-tested yet. We examined variation in growth traits—tree height at ages 4 and 8 years, and diameter at age 13 years—in the common garden experiment testing open-pollinated progenies of 31 seed orchards and 5 commercial seed stands (referred to as populations) at 5 locations. We also examined bud burst phenology at two to five sites at three growing seasons. At one experimental site during the 5th growing season, we measured shoot growth rhythm on all populations. Similar measurements of shoot growth were done on a subset of populations during the 6th growing season together with the analysis of needle growth and foliar chemistry. We found significant variation among populations in growth traits, but also significant population × site (G × E) interactions. We used the regression approach and ecovalence analysis to examine populations’ performance stability. Most populations had average responsiveness to environment, and a set of least-responsive poor-growing populations contributed the most to the G × E interaction. Variation in bud burst phenology was associated with geographical distribution of tested progenies. The early bud-bursting populations originated from the north-eastern to north-central Poland, and a group of late bud-bursting populations originated mainly from the south-eastern region. Correlations between bud burst and growth traits were weak to medium and varied by site, but early bud-bursting populations tended to show stronger growth on height and diameter. We found significant differences among populations in final leader length, shoot elongation time and relative growth rate (RGR). However, RGR and shoot elongation time explained less than 30% of variation in leader length and were weakly correlated with tree height. Populations varied in needle length, specific leaf area and foliar nitrogen concentration, but time trends in these traits did not vary among populations or predefined groups of populations. Therefore, the analysis of growth rhythm or needle traits did not help resolve variation in tree growth to support selection decisions. Contrary to our expectation, progeny of seed orchards did not perform significantly better than that of commercial seed stands. This finding, however, should not be extrapolated beyond our set of populations. Nonetheless, the local seed sources were not always the best. From a selection standpoint, our results help culling the worst populations rather than selecting the best ones. Therefore, testing individual family progeny and genetic roguing of existing seed orchards is highly recommended.     

Evolution and functional impact of rare coding variation from deep sequencing of human exomes

As a first step toward understanding how rare variants contribute to risk for complex diseases, we sequenced 15,585 human protein-coding genes to an average median depth of 111× in 2440 individuals of European (n = 1351) and African (n = 1088) ancestry. We identified over 500,000 single-nucleotide variants (SNVs), the majority of which were rare (86% with a minor allele frequency less than 0.5%), previously unknown (82%), and population-specific (82%). On average, 2.3% of the 13,595 SNVs each person carried were predicted to affect protein function of ~313 genes per genome, and ~95.7% of SNVs predicted to be functionally important were rare. This excess of rare functional variants is due to the combined effects of explosive, recent accelerated population growth and weak purifying selection. Furthermore, we show that large sample sizes will be required to associate rare variants with complex traits.     

Elastic domains regulate growth and organogenesis in the plant shoot apical meristem

Although genetic control of morphogenesis is well established, elaboration of complex shapes requires changes in the mechanical properties of cells. In plants, the first visible sign of leaf formation is a bulge on the flank of the shoot apical meristem. Bulging results from local relaxation of cell walls, which causes them to yield to internal hydrostatic pressure. By manipulation of tissue tension in combination with quantitative live imaging and finite-element modeling, we found that the slow-growing area at the shoot tip is substantially strain-stiffened compared with surrounding fast-growing tissue. We propose that strain stiffening limits growth, restricts organ bulging, and contributes to the meristem's functional zonation. Thus, mechanical signals are not just passive readouts of gene action but feed back on morphogenesis.