BACKGROUND:This study was designed to investigate the impact of bispectral index (BIS) or A-line AAI index (based on middle-latency auditory evoked potential) monitoring on recovery times and drug consumption when compared with standard anaesthetic practice during desflurane-remifentanil anaesthesia.METHODS:After having obtained approval from the institutional review board and written informed consent, 200 adult patients undergoing minor surgical procedures were randomized to receive a desflurane-remifentanil anaesthetic controlled either solely by clinical parameters or by BIS or AAI to the following target values: during maintenance of anaesthesia to a value of '50' (BIS) or '30' (AAI), 15 min before the end of surgery to '60' (BIS) or '45' (AAI). Recovery times and drug consumption were recorded by a blinded investigator.RESULTS:Compared with standard practice, patients with BIS or AAI monitoring needed similar desflurane concentrations (standard practice 2.9 [0.5] vol%, BIS 3.3 [0.9] vol%, AAI 2.6 [0.5] vol%), and had similar recovery times (open eyes 5.6 [2.5] min, 5.9 [3.4] min, 5.0 [3.1] min; extubation 6.3 [2.4] min, 6.6 [3.5] min, 5.6 [3.0] min; stating name 7.3 [2.4] min, 7.6 [3.5] min, 7.3 [6.6] min).CONCLUSIONS:Compared with standard anaesthetic practice BIS and AAI guided titration to the used target ranges did not result in a reduction of desflurane consumption or recovery times during minor surgery with use of remifentanil.

series of 12 bovine pancreatic trypsin inhibitor variants mutated in the P-4 and P-3 positions of the canonical binding loop containing additional R15R and M52L mutations mere used to probe the role of single amino acid substitutions on binding to bovine trypsin and to the following human proteinases involved in blood clotting: plasmin, plasma kallikrein, factors X-a and XIIa, thrombin, and protein C. The mutants were expressed in Escherichia coli as fusion proteins with the LE1413 hydrophobic polypeptide and purified from inclusion bodies; these steps were followed by CNBr cleavage and oxidative refolding, The mutants inhibited the blood-clotting proteinases with association constants in the range of 10(3)-10(10) M-1. Inhibition of plasma kallikrein, factors X-a and XIIa, thrombin, and protein C could be improved by up to 2 orders of magnitude by the K15R substitution The highest increase in the association constant for P-3 mutant was measured for factor XIIa; P13S substitution increased the K-a value 58-fold, Several other substitutions at P-3 resulted in about 10-fold increase for factor X-a, thrombin, and protein C. The cumulative P-3 and P-1 effects on K-alpha values for the strongest mutant compared with the wild type bovine pancreatic trypsin inhibitor were in the range of 2,2- (plasmin) to 4,000-fold (factors XIIa and X-a). The substitutions at the P-4 site always caused negative effects (a decrease in the range from over 1.000- to 1.3-fold) on binding to all studied enzymes, including trypsin, Thermal stability studies showed a very large decrease of the denaturation temperature (about 22 degreesC) for all P-4 mutants, suggesting that substitution of the mild type Gly-12 residue leads to a change in the binding loop conformation manifesting itself in non-optimal binding to the proteinase active site.

Senescence in plants is a complex trait, which is controlled by both genetic and environmental factors and can affect the yield and quality of cotton. However, the genetic basis of cotton senescence remains relatively unknown. In this study, we reported genome-wide association studies (GWAS) based on 185 accessions of upland cotton and 26,999 high-quality single-nucleotide polymorphisms (SNPs) to reveal the genetic basis of cotton senescence. To determine cotton senescence, we evaluated eight traits/indices. Our results revealed a high positive correlation (r>0.5) among SPAD value 20 days after topping (SPAD20d), relative difference of SPAD (RSPAD), nodes above white flower on topping day (NAWF0d), nodes above white flower 7 days after topping (NAWF7d), and number of open bolls on the upper four branches (NB), and genetic analysis revealed that all traits had medium or high heritability ranging from 0.53 to 0.86. Based on a multi-locus method (FASTmrMLM), a total of 63 stable and significant quantitative trait nucleotides (QTNs) were detected, which represented 50 genomic regions (GWAS risk loci) associated with cotton senescence. We observed three reliable loci located on chromosomes A02 (A02_105891088_107196428), D03 (D03_37952328_38393621) and D13 (D13_59408561_60730103) because of their high repeatability. One candidate gene (Ghir_D03G011060) was found in the locus D03_37952328_38393621, and its Arabidopsis thaliana homologous gene (AT5G23040) encodes a cell growth defect factor-like protein (CDF1), which might be involved in chlorophyll synthesis and cell death. Moreover, qRT-PCR showed that the transcript level of Ghir_D03G011060 was down-regulated in old cotton leaves, and virus-induced gene silencing (VIGS) indicated that silencing of Ghir_D03G011060 resulted in leaf chlorosis and promoted leaf senescence. In addition, two candidate genes (Ghir_A02G017660 and Ghir_D13G021720) were identified in loci A02_105891088_107196428 and D13_59408561_60730103, respectively. These results provide new insights into the genetic basis of cotton senescence and will serve as an important reference for the development and implementation of strategies to prevent premature senescence in cotton breeding programs.

The role of the S(1) subsite in trypsin, chymotrypsin and plasmin has been examined by measuring the association with seven different mutants of bovine pancreatic trypsin inhibitor (BPTI); the mutants contain Gly, Ala, Ser, Val, Leu, Arg, and Trp at the P(1) position of the reactive site. The effects of substitutions at the P(1) position on the association constants are very large, comprising seven orders of magnitude for trypsin and plasmin, and over five orders for chymotrypsin. All mutants showed a decrease of the association constant to the three proteinases in the same order: Ala>Gly>Ser>Arg>Val>Leu>Trp. Calorimetric and circular dichroism methods showed that none of the P1 substitutions, except the P1-Val mutant, lead to destabilisation of the binding loop conformation. The X-ray structure of the complex formed between bovine beta-trypsin and P(1)-Leu BPTI showed that the P(1)-Leu sterically conflicts with the side-chain of P(3)-Ile, which thereby is forced to rotate approximately 90 degrees. Ile18 (P(3)) in its new orientation, in turn interacts with the Tyr39 side-chain of trypsin. Introduction of a large side-chain at the P1' position apparently leads to a cascade of small alterations of the trypsin-BPTI interface that seem to destabilise the complex by it adopting a less optimized packing and by tilting the BPTI molecule up to 15 degrees compared to the native trypsin-BPTI complex.