Effects of introduced aspartic and glutamic acid residues on the P'1 substrate specificity, pH dependence and stability of carboxypeptidase Y

Carboxypeptidase Y is a serine carboxypeptidase isolated from Saccharomyces cerevisiae with a preference for C-terminal hydrophobic amino acid residues. In order to alter the inherent substrate specificity of CPD-Y into one for basic amino acid residues in P'1, we have introduced Asp and/or Glu residues at a number of selected positions within the S'1 binding site. The effects of these substitutions on the substrate specificity, pH dependence and protein stability have been evaluated. The results presented here demonstrate that it is possible to obtain significant changes in the substrate preference by introducing charged amino acids into the framework provided by an enzyme with a quite different specificity. The introduced acidic amino acid residues provide a marked pH dependence of the (kcat/Km)FA-A-R-OH/(kcat/Km)FA-A-L-OH ratio. The change in stability upon introduction of Asp/Glu residues can be correlated to the difference in the mean buried surface area between the substituted and the substituting amino acid. Thus, the effects of acidic amino acid residues on the protein stability depend upon whether the introduced amino acid protrudes from the solvent accessible surface as defined by the surrounding residues in the wild type enzyme or is submerged below.     

The dynamics of market-level change

Health care exhibits a competitive dynamic today that increasingly resembles that in other service industries. Organizations are becoming larger to achieve scale economies and to increase market power. Vertical integration, whether through ownership or complex contracts, is also being pursued both to seek efficiencies and to improve the bargaining position of the organization. External forces that are driving these changes include more aggressive activities on the part of purchasers to contain their costs, developments in information technology, management innovation in other service industries, and advances in medical technology. Within the health care industry, there is a pattern of organizations taking the initiative to respond to these external forces--often in anticipation of them--and other organizations then responding to the pressures in turn placed on them. Although information on strategies is communicated rapidly throughout the country, what is attempted and what succeeds differs a great deal across communities. The nature of current health care institutions in the community, including the presence of large entities with extensive capital and strong management in a particular segment of the health system and the community's experience with managed care are important factors in the path that change takes.     

Comfort limits for asymmetric thermal radiation

Groups of 32 and 16 subjects of both sexes were exposed in an environmental chamber to radiant asymmetry caused by a cool wall, a warm wall, and a cool ceiling. Each subject was tested individually while seated and clothed at 0.6 clo. During each 3.5-hour experiment the subject was exposed to six radiant temperature asymmetries. He was asked whether and where he experienced any local cool or warm sensation, and whether it was felt to be uncomfortable. During the entire experiment he was kept thermally neutral by changing the air temperature according to his wishes.For cool walls, warm walls, and cool ceilings curves have been established showing the percentage of dissatisfied subjects as a function of the radiant asymmetry. Radiant asymmetry at a warm wall caused less discomfort than a cool wall. A cool ceiling caused less discomfort than a warm ceiling. Accepting that 5% of the subjects may feel uncomfortable. a radiant temperature asymmetry of 10°C is allowable at a cool wall, 23°C at a warm wall, and 14°C under a cool ceiling. A previous study showed that 4°C is allowable under a warm ceiling. Radiant asymmetry had no significant impact on the operative temperatures preferred by the subjects. No significant differences were observed between the responses of men and women exposed to radiant asymmetry.     

Power management in networked devices

Networks are one of the most significant developments in computing and a hallmark of modern society. However, along with increasing efficiency and productivity, both at home and in the workplace, networks have costs. One cost is the additional energy that electronic devices consume when attached to networks. Power management, a standard feature of modern PCs, was primarily developed to increase battery lifetime in laptop PCs, which historically were not network-connected when using battery power. Today, however, many laptops are connected to a network - typically a Wi-Fi network - as are the majority of desktop computers. Three key drivers of energy use are induced consumption by devices prevented by network connections from entering low-power states, increasing link data rates that inherently consume more energy for the network interfaces, and proliferation of network-connected displays that actively update and display data when no one is present.     

Reflex jaw motions and jaw stiffness pertaining to whiplash injury of the neck

Because a so-called mandibular whiplash injury requires the absence of short-latency jaw-closing reflexes in order to explain the postulated mechanism of injury (excessive jaw opening); the authors studied the presence and absence and more importantly, the kinematics (duration, displacement, velocity, acceleration) of monosynaptic and possibly, polysynaptic myotatic (stretch) reflexes in the jaw elevator muscles. In six healthy adults jaw jerk maneuvers were elicited through a brisk tap on the chin, and surface electromyography identified elevator reflexes while translational electrognathography identified the kinematics of the reflexes. The maneuvers were done while maintaining the rest position (3% MVC) and moderate clenching of the teeth (30% MVC). Electromyography was also used to identify phasic elevator excitations during a passive brisk neck extension maneuver. A sudden and unexpected elongation of the jaw elevators released autogenic reflex responses that, in conjunction with augmented tissue elasticity (stiffness), elevated the mandible into centric occlusion within approximately 150 milliseconds. In 86% of trials, the responses occurred regardless of the prevailing resting and clenching contractile activities. There was no evidence of a depressor force that consistently would and could anchor the mandible in a position of extreme or moderate depression, the theoretical linchpin of the mandibular whiplash injury. It was concluded that the mandibular locomotor system is very efficient in maintaining the rest and intercuspal positions of the mandible. This study found no evidence corroborating the mechanism claimed to release a so-called mandibular whiplash injury.     

Status of risk factors for dementia associated with stroke

This study presents a brief investigation into sex differences of speakers in the voice onset time of English plosives that are stressed in both word-initial and prevocalic position. 72 short phrases were presented to 5 men (range 25 to 37 years, mean age 34.2 yr.) and five women speakers (range 28 to 38 years, mean 32.6 yr.). Analysis showed that the women as speakers had on average, longer voice onset time values than their male peers.     

Role of newly synthesized cholesterol or its metabolites on the regulation of bile acid biosynthesis after short-term biliary diversion in the rat

Cholesterol 7 alpha-hydroxylase, the rate-limiting enzyme in the bile acid biosynthetic pathway, is thought to be regulated by hydrophobic bile acids through negative feedback control. The role of cholesterol in the regulation of cholesterol 7 alpha-hydroxylase is more controversial, in part because of incomplete understanding of the relationship between the pathways of cholesterol synthesis and degradation. The main objective of this study was to define the interaction between these two pathways in an experimental model in which the supply of newly synthesized cholesterol was interrupted by sustained infusion of mevinolin (lovastatin), an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) or accelerated by a continuous infusion of mevalonate, a cholesterol precursor. The study was carried out in rats subjected to short-term bile fistula. In one set of experiments, rats were treated postoperatively with mevinolin (5 mg/kg loading dose followed by 2 mg/kg/hr infusion), mevalonate (180 mumol/hr infusion) or both for up to 96 hr. In a separate set of experiments, rats were infused intraduodenally with taurocholate (36 mumol/100 gm/hr for up to 96 hr). We determined cholesterol 7 alpha-hydroxylase- and HMG-CoA reductase specific activities at those time intervals, whereas bile acid synthesis rates were determined throughout the study. Compared with rats not subjected to surgery, rats with short-term biliary diversion had increases in cholesterol 7 alpha-hydroxylase activity of 259% and 827% at 48 and 96 hr, respectively. The increase in bile acid biosynthesis was less pronounced. Continuous infusion of mevinolin completely prevented increases in cholesterol 7 alpha-hydroxylase specific activity and bile acid biosynthesis at both time intervals.(ABSTRACT TRUNCATED AT 250 WORDS)