Capillary electrochromatography-laser-induced fluorescence method for separation and detection of dansylated dialkylamine tags in encoded combinatorial libraries.

LC-fluorescence and LC-MS methods have been previously reported for use in decoding bead-based combinatorial libraries. We present the use of capillary electrochromatography (CEC) for highly selective decoding in combination with laser-induced fluorescence (LIF) detection for high sensitivity. The results are compared to prior data obtained using HPLC with fluorescence detection. The use of CEC shows promise for miniaturization and multiplexing for future applications, and the use of LIF detection can allow for detection at sub-pmol amounts.

On-line sample preparation for high throughput reversed-phase LC/MS analysis of combinatorial chemistry libraries.

An on-line sample preparation method utilizing a time-programmed autosampler is described for high throughput liquid chromatography/mass spectrometry (LC/MS). This approach is particularly helpful for the LC/MS analysis of samples which require solvents incompatible with HPLC in the sample preparation process. The on-line sample preparation approach minimizes a bottleneck in throughput and improves sample recovery under some circumstances.

Law and the confinement of psychopaths.

Civil commitment in the United States historically has followed a medical, targeting individuals with serious mental disorders and providing for their treatment in the least restrictive setting. In the last decade, however, commitment laws have appeared in some states permitting the hospitalization of personality-disordered criminal offenders at the end of a penal sentence. The American Psychiatric Association has fiercely opposed these laws. The U.S. Supreme Court has given its qualified approval, although legal challenges persist. These laws, together with British proposals to permit the civil commitment of dangerous personality-disordered individuals, should be resisted by all professional disciplines.

Expediting the method development and quality control of reversed-phase liquid chromatography electrospray ionization mass spectrometry for pharmaceutical analysis by using an LC/MS performance test mix.

Mass spectrometry combined with liquid chromatography (LC/MS) has become an important analytical methodology in both pharmaceutical and biomolecule analyses. LC/MS, especially with reversed-phase HPLC (RP-LC), is extensively used in the separation and structural identification of pharmaceutical samples. However, many parameters have to be considered when a new LC/MS method is developed for either separation and structural analysis of unknown mixtures or quantitative analysis of a set of known compounds in an assay. The optimization of a new LC/MS method can be a time-consuming process. A novel kit-LC/MS performance test mix-composed of aspartame, cortisone, reserpine, and dioctyl phthalate has been developed to accelerate the process of establishing a new RP-LC/MS method. The LC/MS mix makes the evaluation and validation of an LC/MS method more efficient and easier. It also simplifies the quality control procedure for an LC/MS method in use.

A novel approach to high-throughput quality control of parallel synthesis libraries.

Combinatorial chemistry is a powerful tool to enhance drug discovery efforts in the pharmaceutical industry. One type of combinatorial chemistry, parallel synthesis, is now widely used to prepare numerous compounds of structural diversity. A novel high-throughput method for quality control of parallel synthesis libraries has been developed. The method uses flow injection MS, for proof of structure and estimation of purity, and a novel direct injection CLND technique for quantitation of amount. Following the synthesis of a small molecule library, compounds analyzed using this technique were characterized by mass spectrometry, and an accurate concentration of the compound was assessed by CLND. Characterization of one compound is completed in 60 s, allowing for up to 1000 compounds to be analyzed in a single day. The data is summarized using pass/fail criteria using internally developed software.

Analytical methods for quality control of combinatorial libraries.

This literature review covers the applications of analytical techniques to solid phase organic chemistry and combinatorial chemistry published between June 96 and September 1997. Highlighted are mass spectrometry, NMR, IR and chromatographic analyses of solid phase synthesis reactions and combinatorial libraries.

Forensic mental health clinical evaluation: an analysis of interstate and intersystemic differences.

Forensic mental health evaluation systems have undergone major changes during the past two decades, and the variability of service delivery systems across states is significant. We compared assessments of competence to stand trial and criminal responsibility in three states with different systems for forensic mental health evaluations: Michigan, Ohio, and Virginia. Although all three states use comparable legal criteria to judge competence and criminal responsibility, we found large, statistically significant differences among the states in the proportion of defendants referred for evaluation who were assessed as incompetent or not criminally responsible. In addition, significant differences were found in the diagnostic and offense categories of defendants referred for evaluation. Our findings suggest that the structure of a system for providing forensic evaluation services may significantly affect both the group of individuals referred for evaluation as well as evaluation outcome.

New methods for analyzing compounds on polymeric supports.

Beyond specialized applications in peptide and oligonucleotide synthesis, widespread utilization of solid-phase methods in organic chemistry has been hampered by a lack of powerful analytical methods for characterization of polymer-supported compounds. The advent of combinatorial organic synthesis has recently spawned efforts to develop spectroscopic techniques such as infrared spectrometry, mass spectrometry and nuclear magnetic resonance as routine tools for structure elucidation in solid-phase synthesis.

Beyond competence and sanity: the influence of pretrial evaluation on case disposition.

A preliminary investigation of the impact of pretrial evaluations of trial competence and legal insanity, and the variables that mediate case outcomes is reported. Twenty-four percent of defendants evaluated as incompetent to stand trial were found competent by the court or were tried without the question of competence being adjudicated. Charges were dropped in more than half of the cases in which an evaluator considered a defendant incompetent, most frequently in cases involving misdemeanor charges and/or the clinician considered it unlikely that the defendant could be restored to competence. One third of defendants considered to meet criteria for legal insanity subsequently were acquitted NGRI; more than half did not present an insanity defense and were ultimately convicted or plea bargained a guilty verdict. The defendants considered to meet the criteria for legal insanity were more likely than their mentally ill but not insane counterparts to have treatment ordered in lieu of incarceration. The defendant's age and race and the evaluator's professional discipline were unrelated to case outcome.

Informed decision making in persons acquitted not guilty by reason of insanity.

Deciding to raise an insanity defense carries serious consequences. This is especially true for persons charged with minor offenses, for whom an acquittal not guilty by reason of insanity (NGRI) might lead to a longer period of incarceration than would conviction. Before raising an insanity defense, a defendant should be provided with information necessary to make an informed decision and should be competent to understand the consequences of the verdict. This study attempted, through retrospective review and concurrent evaluation, to determine the degree to which trial courts in Virginia attended to these important aspects of informed decision making before finding defendants charged with misdemeanors NGRI. The study also attempted to assess the degree to which defendants were competent and informed at the time of adjudication. In most instances, trial courts did not consider defendants' competence to make decisions regarding the insanity defense and did not consider whether defendants were informed about the consequences of a successful insanity defense at the time of adjudication. The average length of stay for these patients was (at least) 21 months; most would have been released earlier had they been committed civilly rather than committed as a result of insanity pleas. We stress the need to educate judges, attorneys and forensic evaluators to the importance of considering defendants competence to plead insanity and of providing information about the consequences of a successful plea. We also propose that laws be changed to recognize the importance of these elements in the decision making process regarding pleas of insanity.

Criminal offense, psychiatric diagnosis, and psycholegal opinion: an analysis of 894 pretrial referrals.

This article presents the results of a study of 894 criminal defendants referred by Virginia courts for evaluation of competency to stand trial or criminal responsibility. All evaluations were conducted on an outpatient basis by mental health professionals who had received specialized training in forensic evaluation. Findings as to the referral questions posed, the criminal offenses charged, and the clinical diagnoses and psycholegal opinions offered by the evaluators are described. Statistical analyses demonstrate significant relationships between both diagnosis and criminal charge and the psycholegal opinion rendered.

Inheritance of low density lipoprotein subclass patterns in familial combined hyperlipidemia.

The inheritance of low density lipoprotein (LDL) subclass patterns was investigated in 234 members of seven large kindreds with familial combined hyperlipidemia (FCHL), a disorder characterized by elevated LDL cholesterol and/or triglyceride and increased coronary disease risk in families. Analysis of LDL subclasses by nondenaturing gradient gel electrophoresis showed a predominance of large, buoyant LDL particles (pattern A) in 71% of the family members and a predominance of small, dense LDL particles (pattern B) in 29% of family members. Based on complex segregation analysis, pattern B appeared to be inherited as an autosomal trait with either a dominant or an additive mode of inheritance and a small, but significant, multifactorial inheritance component. The proposed allele for pattern B was common (frequency = 0.3), and reduced penetrance was observed among men under age 20 and among women under age 50. These results in these FCHL families are consistent with those from a previously reported population-based sample of families, in which pattern B showed an apparent dominant mode of inheritance. In that study, reduced penetrance was observed for men under age 20 and for premenopausal women, but a somewhat lower allele frequency was found for pattern B (0.25). In the FCHL family members, LDL subclass pattern B was associated with significantly increased plasma levels of apolipoprotein B and triglyceride and decreased high density lipoprotein cholesterol. In comparison with a group of controls, the FCHL family members with pattern A had similar mean triglyceride levels, but higher mean apolipoprotein B. Thus, in families with FCHL, a predominance of small, dense LDL particles appears to be inherited as a common, single-gene trait, which is closely associated with the higher plasma triglyceride levels found in these families. The increased plasma apolipoprotein B levels found in FCHL cannot, however, be accounted for by this proposed locus.

Defective catabolism and abnormal composition of low-density lipoproteins from mutant pigs with hypercholesterolemia.

Metabolic and chemical properties of low-density lipoproteins (LDLs) were studied in a strain of pigs carrying a specific apo-B allele associated with hypercholesterolemia and premature atherosclerosis. LDL mass was significantly greater in mutant than in control pigs (400 +/- 55 mg/dL vs 103 +/- 26 mg/dL), as was LDL cholesterol. When normal and mutant LDLs were injected into the bloodstream of normal pigs, the fractional catabolic rate (FCR) of mutant LDL was about 30% lower than that of control LDL. In mutant pigs, the mean FCRs of mutant and control LDL were similar, although they were much lower than the corresponding FCRs observed in normal pigs. The density profile of LDL particles differed in control and mutant pigs; the peak LDL flotation rate was shifted from S0f = 5.3 +/- 1.9 in controls to a more buoyant 7.4 +/- 0.5 in mutants. The elevation of LDL in the mutants was restricted to the most buoyant LDL subspecies. This subpopulation of mutant LDL was enriched with cholesteryl ester (47% vs 37%) and depleted of triglyceride, relative to LDL of similar density and size in controls. The lipid compositions of the denser LDL subpopulations (rho greater than 1.043 g/mL) were similar in mutants and controls. We conclude that the hypercholesterolemia of these mutant pigs is accounted for by defective catabolism of LDL. The buoyant cholesterol ester enriched LDL subspecies that accumulate in plasma may contribute to the accelerated atherogenesis that occurs in these animals.

Protein turnover and 3-methylhistidine excretion in non-pregnant, pregnant and gestational diabetic women.

Protein turnover was studied in nine non-pregnant (NP) and eight pregnant (P) women. The data from two gestational diabetic (GDM) women are included for comparison. Pregnant women were studied at 30-36 weeks gestation. Whole-body protein turnover, synthesis and catabolism rates were measured using a single dose of 15N-glycine followed by measurement of enrichment of urinary ammonia during the next 10 h. P and NP women had similar rates of protein turnover (4.8 g protein/kg/d) and synthesis (3.8 g protein/kg/d). GDM women appeared to have considerably higher rates for both turnover (5.6 g protein/kg/d) and synthesis (4.7 g protein/kg/d). Normal pregnant women excreted significantly more urinary 3-methylhistidine (3MH) than did non-pregnant women (190 vs 149 mumole/d). Correlation between 3MH excretion and protein catabolism rate approached significance (P = 0.087) in the NP women, but was poorly correlated (P = 0.355) in the P women.

Plasma amino acid, glucose, and insulin responses to moderate-protein and high-protein test meals in pregnant, nonpregnant, and gestational diabetic women.

Test meals providing two levels of dietary protein (13% or 26% of the energy) were fed to eight pregnant (P), nine nonpregnant (NP), and two gestational diabetic (GDM) women. Plasma levels of amino acids were measured at 0 h and 2 h. Glucose and insulin were measured at 0, 1/2, and 2 h after the meals. In the fasting state, P women had significantly lower fasting concentrations of most of the amino acids. After the high-protein meal, rises of arginine, ornithine, and branched-chain amino acids (BCAAs: leucine, isoleucine, and valine), were significantly smaller in P women. Changes in BCAAs were normal in GDM women. P women had greater rises of insulin in response to both test meals than did NP women. This may facilitate increased BCAA uptake from the circulation. Rises in plasma glucose tended to be higher in P than NP women, suggesting that insulin's effects on glucose and BCAA uptake may be mediated separately.

Measurement of urinary 3-methylhistidine with cationic-exchange resin.

A simple method is presented for measurement of urinary 3-methylhistidine (3MH) using a cationic exchange resin treatment followed by colorimetric analysis. Equations are given to correct for the interference by histidine (4.3% by mole) in the colorimetric analysis. This correction is especially important for measurement of urinary 3MH in pregnant women or in other subjects with elevated histidine excretion. Good recovery of added standard and good reproducibility of results are documented. Preliminary data from a study of pregnant women are reported, suggesting an increased excretion of 3MH during pregnancy. Large day-to-day variability of 3MH excretion was observed within subjects. It is recommended that repeated measurements be done on each subject when determining 3MH excretion.

Isolation, identification and quantitation of urinary organic acids.

An application of the HISLIB program for the comparison of gas chromatographic-mass spectrometric profiles of urinary organic acids isolated by extraction and ion-exchange methods is described. Ion-exchange methods are clearly superior to solvent extraction in terms of the variety of compounds isolated. However, the former method has practical difficulties which make solvent extraction more attractive for rapid analyses. For the compounds isolated by both methods, the precision of analysis is similar, with standard deviations of relative concentration in the range 10--30% for most compounds.