Continuous monitoring of a restriction enzyme digest of DNA on a microchip with automated capillary sample introduction

Continuous analysis of a DNA restriction enzyme digest on a microfabricated device is demonstrated with minimal intervention and enhanced time resolution. A 62-base-pair fragment of dsDNA containing a KpnI site was used to demonstrate this process. A capillary was used to transfer sample from a single reaction mix to a microfabricated chip with parallel separation lanes. The 6-carboxyfluorescein-labeled DNA fragments were detected with a CCD camera as they separated in the lanes, which were filled with linear polyacrylamide. The products of the restriction enzyme digest were monitored for up to 60 min at an average sampling rate of 1 injection/46 s, with consecutive injections as short as 1 injection/14 s. The digest was injected directly into the chip, eliminating the need for any sample-handling steps after addition of the enzyme to the reaction mix. The effects of temperature and restriction enzyme concentration were briefly examined, as well. This work shows the potential of this method to provide valuable information about the process of restriction enzyme cleavage.     

Alcohol Consumption Moderates the Link Between Cannabis Use and Cannabis Dependence in an Internet Survey.

The link between cannabis use and cannabis dependence remains poorly understood. Some people use cannabis regularly without signs of dependence; others show dependence despite using less. This study examined alcohol consumption as a moderator of this association. A sample of 476 people (primarily Caucasian men) who used cannabis at least once per week reported their alcohol consumption, cannabis use, and cannabis dependence symptoms in an Internet survey. Regressions revealed significant interactions between measures of cannabis use and alcohol consumption when predicting cannabis dependence. Cannabis use covaried with cannabis dependence, particularly in people who consumed alcohol frequently or in large amounts per week. Despite limitations, these data suggest that alcohol may decrease the safety of cannabis consumption. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Robert H Herman Memorial Award in Clinical Nutrition Lecture, 1997. Mechanisms causing loss of lean body mass in kidney disease

In most cell types, including resting skeletal muscle fibers, internal pH (pHi) is kept constant at a relatively alkaline level. The high pHi is obtained in spite of a chronic acid load resulting from cellular metabolism and passive influx of protons driven by electrochemical forces. Regulation of pHi depends on continuous activity of membrane transport systems that mediate an outflux of H+ (or bicarbonate influx), whereby the acid load is counterbalanced. The transporters involved in muscle pH regulation at rest are the Na+/H+ exchange system as well as the Na+-dependent and Na+-independent Cl- bicarbonate transport systems. The Na+/H+ exchanger seems to be active at resting pHi levels in skeletal muscle. Therefore, pH homeostasis in skeletal muscle most likely involves an equilibrium between counter-directed H+ fluxes. A minor fraction of H+ release during intense exercise is mediated by the Na+/H+ exchanger. The capacity of this system is increased with training and hypoxia in rat skeletal muscle. The dominant acid extruding system associated with intense exercise is the lactate/H+ co-transporter. It has been demonstrated that the capacity of the lactate/H+ co-transporter of rat skeletal muscle is upregulated with training and chronic electrical stimulation, and that it is reduced upon denervation and hindlimb unweighting. Moreover, athletes can have an elevated lactate/H+ co-transport capacity, whereas the thigh muscle of spinal cord-injured individuals has a lower transport capacity than the one of healthy untrained subjects. Thus, it appears that the capacity of the lactate/H+ transporter is affected by the level of muscle activity in both rats and humans. In addition, the rate of H+ release from muscle may also be influenced by capillarization and local blood flow. Finally the resulting pH displacement during acid accumulation is determined by the cellular buffer capacity, which may also undergo adaptive changes.     

The muscarinic M1 agonist xanomeline increases soluble amyloid precursor protein release from Chinese hamster ovary-m1 cells

The functionally selective M1 agonist xanomeline, which is currently undergoing clinical trials as a therapy for Alzheimer's disease, was compared to the muscarinic agonist carbachol for effects on secretion of soluble amyloid precursor protein (APPs) from Chinese hamster ovary cells transfected with the human m1 receptor (CHO-m1). Release of APPs from CHO-m1 cells was increased maximally (4-10 fold) by 100 microM carbachol (EC50 = 11 microM) and by 100 nM xanomeline (EC50 = 10 nM). Stimulation of APPs secretion by xanomeline and carbachol was blocked by preincubation with 1 microM atropine. Carbachol did not stimulate APPs secretion from non-transfected CHO cells. Pilocarpine at 1 mM also increased APPs release. The efficacy of carbachol, xanomeline and pilocarpine for stimulating APPs secretion did not differ significantly. Activation of protein kinase C (PKC) in m1 transfected cell lines by 1 microM phorbol dibutyrate (PDBu) increased APPs release, and this was inhibited 97% by the PKC inhibitor bisindolemalemide. The PKC inhibitor decreased xanomeline and carbachol-stimulated APPs secretion by only 25-30%. These results demonstrate that xanomeline increased APPs release by activation of m1 muscarinic receptors and support the possibility that cholinergic replacement therapy for Alzheimer's Disease may reduce amyloid deposition.     

Research contributions to improving managed health care outcomes.

Managed health care programs are beginning to look to findings from psychotherapy outcome research to help set policy, suggesting the need to consider outcomes research from the standpoint of usability or utility. It also provides an opportunity to integrate science and practice. Considering and applying outcomes in this context requires cooperation between scientists and practitioners, the willingness of each group to set aside guild agenda, and giving up favored but insupportable beliefs and practices on the parts of those in both scientist and practitioner camps. This type of cooperation may have mutual payoffs. This article considers some of the obstacles to this type of sacrifice and evaluates some of the potential costs of cooperating. It also provides perspectives on the new roles of psychological assessment and methods of outcome research that would provide a scientific basis for the function of managed health care. (PsycINFO Database Record (c) 2010 APA, all rights reserved)