Direct demonstration of an adaptive constraint

The role of constraint in adaptive evolution is an open question. Directed evolution of an engineered beta-isopropylmalate dehydrogenase (IMDH), with coenzyme specificity switched from nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide phosphate (NADP), always produces mutants with lower affinities for NADP. This result is the correlated response to selection for relief from inhibition by NADPH (the reduced form of NADP) expected of an adaptive landscape subject to three enzymatic constraints: an upper limit to the rate of maximum turnover (kcat), a correlation in NADP and NADPH affinities, and a trade-off between NAD and NADP usage. Two additional constraints, high intracellular NADPH abundance and the cost of compensatory protein synthesis, have ensured the conserved use of NAD by IMDH throughout evolution. Our results show that selective mechanisms and evolutionary constraints are to be understood in terms of underlying adaptive landscapes.     

Effects of bacterial lipopolysachharide on peripheral leukocytes in the American alligator (Alligator mississippiensis)

Alligators were injected intraperitoneally with four different doses (10, 1.0, 0.1, and 0.01 mg/kg body weight) of a mixture of bacterial lipopolysaccharides (LPS) derived from three different types of bacteria (Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae). Injection of the alligators with the LPS mixture resulted in a dose- and time-dependent increase in total peripheral leukocytes Lymphocytes increased at days 3 and 4 post-injection, and decreased back to baseline levels at day 7 for all doses. Alligators that were not treated, and those injected with pyrogen-free saline, did not exhibit statistically significant changes in total leukocytes during the course of the study. Injection of alligators with 0.5 mg LPS/kg body weight derived from one of three bacterial species revealed that the leukocyte increases observed were not statistically different for all three types of LPS. The animals displayed the same increases in total counts and the levels of all circulating leukocyte types were not different between animals treated with a combination of LPS from all three bacterial species.     

Broad spectrum antimicrobial activity of leukocyte extracts from the American alligator (Alligator mississippiensis)

Leukocytes were isolated from whole blood of wild alligators by differential sedimentation. The leukocytes were disrupted in 5% AcOH and the crude extracts processed by ultrafiltration. The extracts were subjected to solvent exchange (0.1% AcOH) and the fraction that contained macromolecules between 1 and 10 kDa were subjected to further analyses. The acid extracts of the alligator leukocytes exhibited substantial antimycotic activities against six of eight species of Candida yeast tested. In addition, the alligator leukocyte extracts were effective as antimicrobial agents against 10 of 12 bacterial species, and displayed moderate activity against two enveloped viruses (human immunodeficiency virus-1 and herpes simplex virus-1_HF). Kinetic analyses revealed that the antimycotic effects of the leukocyte extract occurred rapidly, with 64% fungal growth inhibition within 3 min of exposure. The molecule(s) responsible for the antimicrobial activities were sensitive to proteases, heat-stable, acid soluble, and in the 1–10 kDa range. These data suggest that alligator leukocytes express cationic peptides that are responsible for their antimicrobial properties.     

2009年韩国家禽业展望

对于韩国的家禽生产商,2008年是极不容易的一年.禽流感和投入成本上升对家禽业造成的损害,2009年应该能有所改善.2009年的形势对韩国的家禽生产商来说是乐观的,2008年市场上出现的困难现在已经基本结束,预计2009年将超过2007年的销售记录.     

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.     

Elementary Ca2+ signals through endothelial TRPV4 channels regulate vascular function

Major features of the transcellular signaling mechanism responsible for endothelium-dependent regulation of vascular smooth muscle tone are unresolved. We identified local calcium (Ca(2+)) signals ("sparklets") in the vascular endothelium of resistance arteries that represent Ca(2+) influx through single TRPV4 cation channels. Gating of individual TRPV4 channels within a four-channel cluster was cooperative, with activation of as few as three channels per cell causing maximal dilation through activation of endothelial cell intermediate (IK)- and small (SK)-conductance, Ca(2+)-sensitive potassium (K(+)) channels. Endothelial-dependent muscarinic receptor signaling also acted largely through TRPV4 sparklet-mediated stimulation of IK and SK channels to promote vasodilation. These results support the concept that Ca(2+) influx through single TRPV4 channels is leveraged by the amplifier effect of cooperative channel gating and the high Ca(2+) sensitivity of IK and SK channels to cause vasodilation.     

B cell receptor signal transduction in the GC is short-circuited by high phosphatase activity

Germinal centers (GCs) generate memory B and plasma cells, which are essential for long-lived humoral immunity. GC B cells with high-affinity B cell receptors (BCRs) are selectively expanded. To enable this selection, BCRs of such cells are thought to signal differently from those with lower affinity. We show that, surprisingly, most proliferating GC B cells did not demonstrate active BCR signaling. Rather, spontaneous and induced signaling was limited by increased phosphatase activity. Accordingly, both SH2 domain-containing phosphatase-1 (SHP-1) and SH2 domain-containing inositol 5 phosphatase were hyperphosphorylated in GC cells and remained colocalized with BCRs after ligation. Furthermore, SHP-1 was required for GC maintenance. Intriguingly, GC B cells in the cell-cycle G(2) period regained responsiveness to BCR stimulation. These data have implications for how higher-affinity B cells are selected in the GC.     

An impactor origin for lunar magnetic anomalies

The Moon possesses strong magnetic anomalies that are enigmatic given the weak magnetism of lunar rocks. We show that the most prominent grouping of anomalies can be explained by highly magnetic extralunar materials from the projectile that formed the largest and oldest impact crater on the Moon: the South Pole-Aitken basin. The distribution of projectile materials from a model oblique impact coincides with the distribution of magnetic anomalies surrounding this basin, and the magnetic properties of these materials can account for the intensity of the observed anomalies if they were magnetized in a core dynamo field. Distal ejecta from this event can explain the origin of isolated magnetic anomalies far from this basin.