Antisense-induced reduction in nucleus accumbens cyclic AMP response element binding protein attenuates cocaine reinforcement.

Repeated cocaine exposure up-regulates cyclic AMP signaling and increases the transcriptional activity of cyclic AMP response element binding protein (CREB) in the nucleus accumbens. To study the possibility that nucleus accumbens CREB activity regulates self-administration behavior, we tested the effects of a single, bilateral infusion of CREB antisense oligonucleotide into nucleus accumbens core and shell sub-regions on cocaine self-administration in rats. Nucleus accumbens core infusions of CREB antisense reduced CREB and the CREB-regulated immediate early gene brain-derived neurotrophic factor by 31 and 27%, respectively, but failed to alter levels of the homologous CREB family proteins cyclic AMP response element modulator and activating transcription factor 1, and had no effect on CREB levels in adjacent nucleus accumbens shell tissue. Similar infusions of CREB antisense in either core or shell produced a transient downward shift in cocaine self-administration dose-response curves on a fixed ratio 5 (five responses/injection) reinforcement schedule, indicating a reduction in cocaine reinforcement that fully recovered 3 days after treatment. CREB antisense also increased the threshold dose of cocaine required for reinstating cocaine self-administration, indicating that nucleus accumbens CREB levels regulate the incentive properties of cocaine. When access to cocaine was less restricted on a fixed ratio 1 schedule, infusion of CREB antisense in the core, but not shell, caused a transient (1-2 days) reduction in stabilized cocaine self-administration, but had no effect on responding maintained by sucrose pellets, indicating that basal CREB levels in the nucleus accumbens core regulate drug intake. None of these effects were produced by nucleus accumbens infusions of complementary sense oligonucleotide. These results suggest a necessary role for nucleus accumbens CREB activity in cocaine reinforcement, and, by converse analogy, up-regulation in CREB activity after chronic cocaine use could contribute to addiction-related increases in cocaine self-administration.

Role for GDNF in biochemical and behavioral adaptations to drugs of abuse.

The present study examined a role for GDNF in adaptations to drugs of abuse. Infusion of GDNF into the ventral tegmental area (VTA), a dopaminergic brain region important for addiction, blocks certain biochemical adaptations to chronic cocaine or morphine as well as the rewarding effects of cocaine. Conversely, responses to cocaine are enhanced in rats by intra-VTA infusion of an anti-GDNF antibody and in mice heterozygous for a null mutation in the GDNF gene. Chronic morphine or cocaine exposure decreases levels of phosphoRet, the protein kinase that mediates GDNF signaling, in the VTA. Together, these results suggest a feedback loop, whereby drugs of abuse decrease signaling through endogenous GDNF pathways in the VTA, which then increases the behavioral sensitivity to subsequent drug exposure.

Expression of the transcription factor deltaFosB in the brain controls sensitivity to cocaine.

Acute exposure to cocaine transiently induces several Fos family transcription factors in the nucleus accumbens, a region of the brain that is important for addiction. In contrast, chronic exposure to cocaine does not induce these proteins, but instead causes the persistent expression of highly stable isoforms of deltaFosB. deltaFosB is also induced in the nucleus accumbens by repeated exposure to other drugs of abuse, including amphetamine, morphine, nicotine and phencyclidine. The sustained accumulation of deltaFosB in the nucleus accumbens indicates that this transcription factor may mediate some of the persistent neural and behavioural plasticity that accompanies chronic drug exposure. Using transgenic mice in which deltaFosB can be induced in adults in the subset of nucleus accumbens neurons in which cocaine induces the protein, we show that deltaFosB expression increases the responsiveness of an animal to the rewarding and locomotor-activating effects of cocaine. These effects of deltaFosB appear to be mediated partly by induction of the AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole) glutamate receptor subunit GluR2 in the nucleus accumbens. These results support a model in which deltaFosB, by altering gene expression, enhances sensitivity to cocaine and may thereby contribute to cocaine addiction.

A model for selecting a clinical laboratory information system. A four-phase process.

Laboratory information systems (LISs) have become an essential part of an efficient and effective laboratory. In the past, selecting an LIS was a relatively simple procedure because there were only a few candidates to select from. Now, however, selecting an LIS from the myriad available has become a time-consuming and complex process. This article presents a multi-attribute utility (MAU) method for selecting an LIS. This MAU method has four phases: identifying LIS vendors and disseminating requests for proposals (RFPs), analyzing RFP responses and selecting the top three vendors, validating responses of the top three vendors, and selecting a primary vendor and preparing a formal recommendation. By following this four-phase process, laboratories will simplify the complex LIS selection process and increase their chances of selecting the best LIS for their needs.

A multicenter evaluation of the Boehringer Mannheim/Hitachi 717 system.

Analytical performance of the Boehringer Mannheim/Hitachi 717 system was evaluated in a multicenter study involving seven different laboratories. Fifty-five methods including end point chemistries, enzymes, ISE, TDM, DAU, and specific protein assays were assessed over a 7 month period. Methods on the analyzer exhibited excellent precision with CVs less than 2% for within run precision, and CVs less than 3% for between day precision for most analytes; linearity, which met or exceeded manufacturer's claims; minimal sample and reagent carryover, and no significant interference from hemolysis; icterus; and lipemia. Recovery of the assigned value for 10 analytes in SRM 909 was acceptable. Comparison of methods with other BM/Hitachi analyzers resulted in slopes close to unity (0.93-1.06); comparison to other clinical chemistry analyzers yielded slopes of 0.88-1.07. Excellent performance and diverse method applications make the BM/Hitachi 717 analyzer a suitable instrument for work station consolidation.

Automated sample handling in a clinical laboratory.

Bar codes have received wide acceptance and have permeated many industries since their early days in the railroad industry. They offer a simple and cost-effective approach to a variety of operational situations. Clinical labs are especially well-suited for this technology.

Test strip meconium screening for cystic fibrosis.

The Boehringer-Mannheim Corporation (BMC) strip test is extremely reliable in indicating an albumin content above 20 mg/gm of dried meconium. All infants born during one year in 14 Milwaukee area hospitals were tested. Of 16,224 newborns, two were diagnosed correctly as suffering from cystic fibrosis and two were missed. False-positive tests were obtained in 0.9% of infants (prematurity, melena, gastroschisis, and intrauterine infection). The strip test is, at present, the best available but not the perfect screening method for cystic fibrosis.