Determination of drug levels in two species of necrophagous Coleoptera reared on substrates containing morphine

Two species of necrophagous Coleoptera: Dermestes frischi (Dermestidae) and Thanatophilus sinuatus (Silphidae), were reared on substrates containing different amounts of morphine. Colonies of D. frischi were reared on rabbit carcasses which had been given 10, 20, and 40 mg/h of morphine hydrochloride via ear artery perfusion over a 3 h period prior to death. A fourth rabbit served as a control. T. sinuatus was reared on minced beef spiked with morphine hydrochloride to give concentrations of 1,000, 2,500, 5,000, and 10,000 ng/g and one control colony. These dosages were calculated to create tissue concentrations of morphine similar to those encountered in human deaths due to morphine overdose. Larvae. pupae, and adults (except for T. sinuatus) were analyzed for morphine content. All developmental stages of D. frischi were positive for morphine and concentrations correlated with cadaveric tissue concentrations during larval stages and to a lesser extent in the adult stage. For T. sinuatus, the best correlations were found in 2nd and 3rd instar larvae. This study demonstrates the potential for use of necrophagous Coleoptera, as well as Diptera larvae, as alternate specimens for toxicological analyses.     

A standard of AmpliType PM typing from aged evidentiary samples

In analyzing aged samples by the AmpliType PM PCR amplification and Typing kit, it was occasionally observed that color developed typing strips had dark allele dots on PM loci but no visible S dot. Since the S dot acts as a minimum dot intensity control to determine positive alleles on the PM loci, it is necessary to apply another control system. To achieve positive PM typing from a degraded DNA sample that is inferred to be derived from a single donor, a standard has been adopted wherein loci from which sufficient PCR products are observed on agarose gel can be typed. The objective determination of sufficient PCR was done by comparison between band peak height of each locus generated from a sample and that of the corresponding locus generated from two nanograms (recommended minimum quantity as template DNA) of the control DNA provided in the kit.     

The structure, frequency, and forensic application of the STR locus D16S543 in the Japanese population

D16S543 is a complex STR locus consisting of five types of repeat units. The frequency distribution and genetic characteristics of this locus in Japanese were investigated using blood samples from 124 unrelated Japanese and 15 families. Alleles were detected using denatured polyacrylamide gels followed by automated analysis on an ABI 373 sequencer using Genescan software 672. Twenty-one alleles were identified, ranging in size from 281 to 489 bp. An allelic ladder containing the 21 alleles was constructed and used as a typing standard. The repeat unit arrays allowed the 21 alleles to be classified into three distinct groups, including alleles 1 to 7 in group I, alleles 8 to 14 in group II, and alleles 15 to 22 in group III. The alleles in group II were characterized by the insertion of one repeat unit of CAGG, one of AAAG, and three of AAGG, while the group III alleles differed from those of groups I and II by the insertion of a total of 32 repeat units ranging in 5 types. Within each group, the alleles differed from each other only in one 5' side tetranucleotide AAGG. The power of discrimination (Pd) and the estimated heterozygosity were calculated to be 0.989 and 0.934, respectively. Typing of this locus was successfully applied in four old forensic materials. The study presented herein demonstrates that D16S543 is a highly polymorphic and applicable locus in Japanese.     

Demonstration and interpretation of bridging vein ruptures in cases of infantile subdural bleedings

Report of two cases of lethal infantile subdural bleedings (SDB). Bridging vein (BV) ruptures were directly proven as the source of the (minimal) SDB by a postmortem X-ray. In the controversial discussion concerning the causes of infantile SDB, proof of the occurrence of several BV ruptures is seen as an important sign of a trauma of significant degree. Although infantile SDB undoubtedly can result from accidental as well as intentional injuries, and therefore, the SDB itself does not allow far-reaching conclusions as to the cause of injury, the presence of several BV ruptures combined with an SDB of insignificant volume, in an infant dead or in a deep coma on clinical presentation, is not compatible with the supposition of a minor fall as the cause. We have not observed such findings as the result of a minor accidental event for more than 15 years.     

An analytical method for the rapid screening of organophosphate pesticides in human biological samples and foodstuffs

Gas chromatography with nitrogen/phosphorus sensitive detection (GC/PND) and electron impact mass spectrometry (GC/MS) with selected ion monitoring provides a simple, rapid and sensitive method for the determination of organophosphate pesticides (OPs). A selective single-step extraction of 23 different OPs in urine, blood, serum and food samples (baby food, soft drinks and instant soups suspected of contamination from a blackmailing scare) is described. The OPs were extracted with 1ml toluene (with and without addition of mevinphos as internal standard), using a 0.7ml aliquot of urine, blood or serum sample. Food samples (0.2g) were homogenised with water (0.5ml) before extraction. An amount of 1microl of the toluene phase (extraction supernatant) was analysed directly by GC/PND and GC/MS.The method was validated using spiked human serum. The OPs were mixed with serum containing 10mg/ml disodium ethane diamine tetraacetic acid disodium salt (EDTA disodium salt) and stored up to 10 days at 4 and -20 degrees C, respectively. The recovery rates of OPs in freshly spiked human plasma ranged between 50% (dimethoate) and 133% (dialifos). OPs in plasma proved to be stable at -20 degrees C. Their levels decreased only slightly after storage at 4 degrees C.     

Detection of the cholinesterase inhibitor tacrine and its main metabolites

Tacrine, a cholinesterase inhibitor for symptomatic treatment of minor to moderate dementia, and its primary metabolites 1-hydroxy-tacrine and 4-hydroxy-tacrine were studied by means of thin-layer chromatography, UV spectroscopy and gas-chromatography/mass spectroscopy. The analytical data (corrected hRf values, UV spectra in solution as well as reflectance spectra, high-pressure liquid chromatography data, GC retention indices and EI mass spectra) including various derivatization methods are described.     

N-cyanomethyl-N-methyl-1-(3',4'-methylenedioxyphenyl)-2-propylamine: an MDMA manufacturing by-product

This paper describes the structural elucidation of a compound produced during the synthesis of 3,4-methylenedioxymethylamphetamine (MDMA) via the reductive amination of 3,4-methylenedioxyphenyl-2-propanone (3,4-MDP-2-P) with methylamine and sodium cyanoborohydride. The compound was isolated from MDMA by column chromatography, proton and carbon nuclear magnetic resonance spectroscopy, LC/mass spectrometry, and total synthesis were used to identify the compound as N-cyanomethyl-N-methyl-1-(3',4'-methylenedioxyphenyl)-2-propylamine. This compound has been identified as a potential synthetic route marker for the reductive amination of 3,4-MDP-2-P with methylamine and sodium cyanoborohydride and as such it should prove valuable to forensic scientists engaged in profiling illicit drugs. Profiling MDMA can provide useful information to law enforcement agencies relating to synthetic route, precursor chemicals and reagents employed and may be used for comparative analyses of different drug seizures. This paper also describes the structural elucidation of the analogous methylamphetamine synthetic route marker compound, N-cyanomethyl-N-methyl-1-phenyl-2-propylamine, produced during the reductive amination of phenyl-2-propanone using methylamine and sodium cyanoborohydride.     

Population genetic analysis of 15 STR loci of Chinese Tu ethnic minority group

We studied and established a DNA database of 15 euchromosome STRs (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA) in a population sample of 151 unrelated individuals of Tu ethnic minority group. Allelic frequencies and statistical parameters of Tu population were calculated. Totally 136 alleles were observed, with the corresponding allelic frequencies ranging from 0.0033 to 0.5359. Chi-square test showed that all STR loci agreed with Hardy-Weinberg equilibrium. Our study population data were compared with the previously publishing population data of other ethnic groups or areas. Our results of present study were valuable for human identification and paternity tests in Chinese Tu population.