Human dopamine transporter gene: coding region conservation among normal, Tourette's disorder, alcohol dependence and attention-deficit hyperactivity disorder populations

The dopamine transporter (DAT) provides major regulation of the synaptic levels of dopamine and is a principal target of psychostimulant drugs. Associations between DAT gene polymorphisms and human disorders with possible links to dopaminergic neurotransmission, including attention-deficit/hyperactivity disorder (ADHD) and consequences of cocaine and alcohol administration, have been reported. We now report approximately 60000 bp of genomic sequence containing the entire DAT gene. This sequence was used to amplify each of the 15 DAT gene exons and several introns and analyze these amplification products by single-stranded sequence conformation (SSCP) and/or direct sequencing. These results define silent allelic single nucleotide sequence variants in DAT gene exons 2, 6, 9 and 15. Rare conservative mutations are identified in amino acids encoded by DAT exons 2 and 8. Analyses of the common nucleotide variants and the previously reported VNTR in the non-coding region of exon 15 define the pattern of linkage disequilibrium across the DAT locus. These comprehensive analyses, however, fail to identify any common protein coding DAT sequence variant in more than 150 unrelated individuals free of neuropsychiatric disease, 109 individuals meeting City of Hope criteria for Tourette's syndrome, 64 individuals with DSM-IV diagnoses of ethanol dependence, or 15 individuals with ADHD. These data are consistent with substantial evolutionary conservation of the DAT protein sequence. They suggest that gene variants that alter levels of DAT expression provide the best current candidate mechanism for reported associations between DAT gene markers, ADHD and other more tentatively associated neuropsychiatric disorders.     

Cholinesterase inhibition by potato glycoalkaloids slows mivacurium metabolism

BACKGROUND: The duration of action for many pharmaceutical agents is dependent on their breakdown by endogenous hydrolytic enzymes. Dietary factors that interact with these enzyme systems may alter drug efficacy and time course. Cholinesterases such as acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) hydrolyze and inactivate several anesthetic drugs, including cocaine, heroin, esmolol, local ester anesthetics, and neuromuscular blocking drugs. Natural glycoalkaloid toxins produced by plants of the family Solanaceae, which includes potatoes and tomatoes, inhibit both AChE and BuChE. Here the authors assess the extent to which two solanaceous glycoalkaloids (SGAs), alpha-solanine and alpha-chaconine, can alter the effects of neuromuscular blocking drugs and cholinesterase inhibitors in vivo and in vitro. METHODS: Inhibition of purified human AChE and BuChE by SGAs, neuromuscular blocking drugs, and cholinesterase inhibitors was assessed by an in vitro colorimetric cholinesterase assay. In vivo experiments were carried out using anesthetized rabbits to test whether SGAs affect recovery from mivacurium-induced paralysis. RESULTS: SGAs inhibited human BuChE at concentrations similar to those found in serum of individuals who have eaten a standard serving of potatoes. Coapplication of SGAs (30-100 nm) with neuromuscular blocking drugs and cholinesterase inhibitors produced additive cholinesterase inhibition. SGA administration to anesthetized rabbits inhibited serum cholinesterase activity and mivacurium hydrolysis. In addition, SGA prolonged the time needed for recovery from mivacurium-induced paralysis (149 +/- 12% of control; n = 12). CONCLUSIONS: These findings support the hypothesis that inhibition of endogenous enzyme systems by dietary factors can influence anesthetic drug metabolism and duration of action. Diet may contribute to the wide variation in recovery time from neuromuscular blockade seen in normal, healthy individuals.     

Resident training in clinical chemistry

Practicing clinical chemists responded to an anonymous, open-ended questionnaire designed to define the state of clinical chemistry education in pathology training programs in the United States. Survey respondents identified many ideas for educational improvements and offered criticism regarding aspects of clinical chemistry education that are not working particularly well. Many of these findings are generalizable to other subspecialties of clinical pathology. It is hoped that this analysis will allow readers to compare their programs with national trends and identify new ways of improving clinical chemistry training at their institutions.     

Surface Finish Analysis in Single Point Incremental Sheet Forming of Rib-Stiffened 2024-T3 and 7075-T6 Alclad Aluminium Alloy Panels

The article presents the results of the analysis of the interactions between the single point incremental forming (SPIF) process parameters and the main roughness parameters of stiffened ribs fabricated in Alclad aluminium alloy panels. EN AW-7075-T6 and EN AW-2024-T3 Alclad aluminium alloy sheets were used as the research material. Panels with longitudinal ribs were produced with different values of incremental vertical step size and tool rotational speed. Alclad is formed of high-purity aluminium surface layers metallurgically bonded to aluminium alloy core material. The quality of the surface roughness and unbroken Alclad are key problems in SPIF of Alclad sheets destined for aerospace applications. The interactions between the SPIF process parameters and the main roughness parameters of the stiffened ribs were determined. The influence of forming parameters on average roughness Sa and the 10-point peak–valley surface roughness Sz was determined using artificial neural networks. The greater the value of the incremental vertical step size, the more prominent the ridges found in the inner surface of stiffened ribs, especially in the case of both Alclad aluminium alloy sheets. The predictive models of ANNs for the Sa and the Sz were characterised by performance measures with R² values lying between 0.657 and 0.979. A different character of change in surface roughness was found for sheets covered with and not covered with a soft layer of technically pure aluminium. In the case of Alclad sheets, increasing the value of the incremental vertical step size increases the value of the surface roughness parameters Sa and Sz. In the case of the sheets not covered by Alclad, reduction of the tool rotational speed increases the Sz parameter and decreases the Sa parameter. An obvious increase in the Sz parameter was observed with an increase in the incremental vertical step size.     

Ultimate Load-Carrying Ability of Rib-Stiffened 2024-T3 and 7075-T6 Aluminium Alloy Panels under Axial Compression

Stringer-stiffened panels made of aluminium alloys are often used as structural elements in the aircraft industry. The load-carrying capacity of this type of structure cannot relieve the reduction in strength in the event of local buckling. In this paper, a method of fabrication of rib-stiffened panels made of EN AW-2024-T3 Alclad and EN AW-7075-T6 Alclad has been proposed using single point incremental forming. Panels made of sheets of different thickness and with different values of forming parameters were tested under the axial compression test. A digital image correlation (DIC)-based system was used to find the distribution of strain in the panels. The results of the axial compression tests revealed that the panels had two distinct buckling modes: (i) The panels buckled halfway up the panel height towards the rib, without any appreciable loss of rib stability, and (ii) the rib first lost stability at half its height with associated breakage, and then the panel was deflected in the opposite direction to the position of the rib. Different buckling modes can be associated with the character of transverse and longitudinal springback of panels resulting from local interaction of the rotating tool on the surface of the formed ribs.     

The Influence of Low-Molecular-Weight Monomers (TEGDMA,HDDMA, HEMA) on the Properties of Selected Matrices and Composites Based on Bis-GMA and UDMA

Bisphenol A-glycidyl methacrylate (bis-GMA) and urethane dimethacrylate (UDMA) are usually combined with low-viscosity monomers to obtain more desirable viscosity, handling characteristics and general properties. The present study determined the flexural strength (FS), flexural modulus (FM), diametral tensile strength (DTS), and hardness (HV) of five matrices and composites based on these resins. The polymerization shrinkage stress (PSS) was also studied for the composites. The polymer matrices were formed using bis-GMA and UDMA. TEGDMA, HEMA and HDDMA acted as co-monomers. The composites had 45 wt.% of filler content. The highest FS and FM were obtained from the UDMA/bis-GMA/TEGDMA/HEMA matrix and the composite (matrix + filler). The best DTS values were obtained from the UDMA/bis-GMA/HEMA matrix and the composite. One of the lowest values of FS, FM, and DTS was obtained from the UDMA/bis-GMA/HDDMA matrix and the composite. All the composites demonstrated similar hardness values. The lowest polymerization shrinkage stress was observed for the UDMA/bis-GMA/TEGDMA/HEMA composite, and the highest PSS was observed for the UDMA/bis-GMA/TEGDMA/HDDMA composite. The addition of HEMA had a positive effect on the properties of the tested materials, which may be related to the improved mobility of the bis-GMA and UDMA monomers.