Genotyping for DQA1 and PM loci in urine using PCR-based amplification: effects of sample volume, storage temperature, preservatives, and aging on DNA extraction and typing.

Urine is often the sample of choice for drug screening in aviation/general forensic toxicology and in workplace drug testing. In some instances, the origin of the submitted samples may be challenged because of the medicolegal and socioeconomic consequences of a positive drug test. Methods for individualization of biological samples have reached a new boundary with the application of the polymerase chain reaction (PCR) in DNA profiling, but a successful characterization of the urine specimens depends on the quantity and quality of DNA present in the samples. Therefore, the present study investigated the influence of storage conditions, sample volume, concentration modes, extraction procedures, and chemical preservations on the quantity of DNA recovered, as well as the success rate of PCR-based genotyping for DQA1 and PM loci in urine. Urine specimens from male and female volunteers were divided and stored at various temperatures for up to 30 days. The results suggested that sample purification by dialfiltration, using 3000-100,000 molecular weight cut-off filters, did not enhance DNA recovery and typing rate as compared with simple centrifugation procedures. Extraction of urinary DNA by the organic method and by the resin method gave comparable typing results. Larger sample volume yielded a higher amount of DNA, but the typing rates were not affected for sample volumes between 1 and 5 ml. The quantifiable amounts of DNA present were found to be greater in female (14-200 ng/ml) than in male (4-60 ng/ml) samples and decreased with the elapsed time under both room temperature (RT) and frozen storage. Typing of the male samples also demonstrated that RT storage samples produced significantly higher success rates than that of frozen samples, while there was only marginal difference in the DNA typing rates among the conditions tested using female samples. Successful assignment of DQA1 + PM genotype was achieved for all samples of fresh urine, independent of gender, starting sample volume, or concentration method. Preservation by 0.25% sodium azide was acceptable for sample storage at 4 degrees C during a period of 30 days. For longer storage duration, freezing at -70 degrees C may be more appropriate. Thus, the applicability of the DQA1 + PM typing was clearly demonstrated for individualization of urine samples.     

GC/MS determination of zolpidem in postmortem specimens in a voluntary intoxication

An ingestion of an unknown quantity of Ivadal (zolpidem) tablets in a case of drug abuse is described. The authors report a new and fast method of analysing and determining the zolpidem concentration in postmortem specimens. Quantitation of zolpidem was performed by ethyl acetate extraction from alkalinized body fluids before GC/MS analysis. The analyses were performed without any complex sample clean-up steps and with little sample material. Postmortem concentrations of zolpidem in body fluids are given. The proposed method is a rapid procedure of analysis in cases of deliberate poisoning with the sedative-hypnotic drug, zolpidem.     

Are Overlapping Local Governments Competing With Each Other When Issuing Debt?

Local public services are produced through various overlapping jurisdictions. This study examines how the issuance of municipal general obligation bonds is affected by the tax policies of overlapping local governments. The findings challenge the hypothesis that the shared tax base would be overused in a common-pool resource scenario. Instead, the empirical results show that the issuance of general obligation bonds is more likely in jurisdictions where counties and school districts make more intensive use of the property tax. These findings highlight the importance of the signals local governments receive from their overlapping neighbors regarding voters’ demand for additional public spending.     

In- and out-flows of elements in bones embedded in reference soils

Possible exchanges of elements between bone and the surrounding soil after being embedded underground for 2 years were estimated. Bone pieces were samples from human vertebrae without any treatments after resection. Sixteen elements were determined by atomic emission mass spectrometry. These were divided into three types; Type I, an in-flow in which elements increased, as in Fe, Al and Ba; type II, a balanced decrease in which changes were found in S, Mg and Zn; and type III, an out-flow in which elements, such as Ca and P, entered into bones from embedded soils. These exchanges depended on the varying nature of soils and also on the time underground. The exchanges were progressed in duration of the time after burial. Data obtained are possible references to judge the time-lapse after burial of bones in relating to characters of soils embedded, and to identify proper bone elements from containment elements.     

First, do no harm: the use of covert video surveillance to detect Munchausen Syndrome by Proxy--an unethical means of "preventing" child abuse

Since it was first identified in 1977, Munchausen Syndrome by Proxy has uniquely affected the way in which the medical and legal communities deal with the issue of child abuse. Inherent in the medical response to the disease are issues of suspicion, investigation, identification, confrontation, and, of course, the health of an innocent child. Given the deceptive dynamics of this disease, however, denial and disbelief naturally overshadow every action taken by medical professionals in pursuing these issues. Fortunately, as medical knowledge about the dynamics of the disease continues to develop, medical professionals become more willing and better able to identify the disease and focus their response on the safety of the child. The greatest problem in prosecuting Munchausen Syndrome by Proxy is that judges and juries remain unwilling to accept the reality of the disease. Consequently, in an effort to confirm medical suspicions and quell legal doubts, the medical community has resorted to covert video surveillance of the abuse while it is being perpetrated in the hospital. In this Article, Flannery argues that this response is an unnecessary and unethical, means of preventing Munchausen Syndrome by Proxy and protecting the child. Flannery supports the approach taken by the Family Court of New York in addressing Munchausen Syndrome by Proxy cases. The Family Court of New York recognizes the unique dynamics of this bizarre disorder, and, therefore, considers all cumulative circumstantial evidence in a Munchausen Syndrome by Proxy case, comparing the facts of the subject case to the commonly accepted features of confirmed cases. Part of the circumstantial evidence that should be considered, Flannery argues, is the dissipation of the child's condition upon temporary separation from the alleged perpetrating parent. As is done by the Flannery Court of New York, a res ipsa loquitur standard should then be applied, and an appropriate disposition for the child should be determined. By employing this standard, the court may confirm suspicions of Munchausen Syndrome by Proxy while avoiding the unnecessary harm to the child inherent in the covert video surveillance of Munchausen Syndrome by Proxy.