Successful extraction of human genomic DNA from serum and its application to forensic identification

We report here on the successful extraction of human genomic DNA from a serum sample in a forensic case. The extracted DNA was successfully used for the identification of remains presumably immersed for more than three weeks for which the only comparison sample was a 250-microL serum aliquot kept frozen in a laboratory. The analysis made it possible to identify a second victim as the daughter of the first.     

Brazilian population data on the polymerase chain reaction-based loci LDLR,GYPA, HBGG,D7S8, and Gc

Gene and genotype frequencies in relation to the low density lipoprotein receptor (LDLR), glycophorin A (GYPA), hemoglobin G gammaglobin (HBGG), D7S8, and group specific component (Gc) loci were determined in a sample of 344 unrelated individuals (250 whites and 94 mulattoes) living in the city of S?o Paulo, Brazil. DNA was extracted from 5 mL of peripheral blood obtained from each of the 344 volunteers by the salting-out procedure. Polymerase chain reaction and reverse dot-blot analysis were performed with the Amplitype PM PCR Amplification and Typing Kit (Polymarker Multiplex; Applied Biosystems, Foster City, CA) under conditions recommended by the manufacturer. Estimated allele frequencies in the white sample were in the usual range of that of other United States and European population groups. In any case, genotype distributions for these loci did not deviate significantly from Hardy-Weinberg equilibrium proportions. Only 1 marginally significant (0.01 < P < 0.05) association, between loci HBGG and Gc, was detected in our mulatto sample out of a total of 20 possible pairwise comparisons of the 5 loci for both data sets. Allele frequencies were significantly different (P < 0.001) at the HBGG and Gc loci when the white and mulatto samples were compared. Biologic relationship exclusion probabilities (test powers) were calculated for the data. A Brazilian database has thus been established for the loci LDLR, GYPA, HBGG, D7S8, and Gc, 5 polymerase chain reaction-based loci systems that have been shown to be a useful tool for biologic relationship identification and exclusion.     

Genetic profiling of a central Venezuelan population using 15 STR markers that may be of forensic importance

The AmpFlSTR Identifiler kit has recently been accepted for use in DNA databasing of forensic samples in the FBI's National DNA Index System. In the present study, we used this kit to analyze the allele distribution of 15 short tandem repeat markers (STR) in individuals living in Caracas city, Venezuela. The allele frequencies of two of these STR, D2S1338 and D19S433, have not previously been reported for this or any other Latin American population. The results indicate that for the population here studied, the 15 STR tested are useful markers for paternity testing and forensic casework.     

Instability of calcium channel antagonists during sample preparation for LC-MS-MS analysis of serum samples

1,4-Dihydropyridines calcium channel antagonists (1,4-DHP CCAs) are photolabile and the products of their photodecomposition have no pharmaceutical activity. In our previous work we have presented a screening procedure for eleven 1,4-DHPs in plasma by LC-MS-MS using multiple reaction motoring. The laboratory process includes preparation and storage of stock solutions, plasma storage, solid-phase extraction, reconstitution of extracts and storage time in an autosampler for LC-MS-MS analysis. Prior to validation of the analytical procedure, we have tested the stability of these compounds by exposure to light. Methanolic solutions have been exposed to laboratory and UV light and the stability of the compounds in plasma was tested by exposure of spiked plasma samples to laboratory light at room temperature. Stability during freeze-thaw cycles and stability during 2 month storage at -20 degrees C have been tested as well. Products of photodecomposition have been identified after forced degradation and the degree of degradation has been quantified using LC-UV-DAD and LC-MS-MS, respectively. A 96% degradation after only 2h has been observed when solutions of nifedipine or nisoldipine were exposed to laboratory light in clear glass vials. In plasma samples degradation was 25% in only 2h for both compounds. The main degradation product was produced by oxidation of the dihydropyridinic ring resulting in the pyridine analogue that has been described as the first metabolite in the metabolic pathway. Only minor degradation was found for the other tested compounds after 2h light exposure in methanolic solutions. Furthermore, lercanidipine and nicardipine were also degradated by esterhydrolysis. Several additional minor degradation products were found for the other tested 1,4-DHPs, however, some of them could not be identified. Preconditions for storage and handling of plasma samples prior to and during analysis for 1,4-DHP CCAs are suggested in order to avoid photodecomposition of the analytes.     

Survival of severe amlodipine intoxication due to medical intensive care

We report the case of attempted suicide with amlodipine, chlorthalidone and mefenamic acid and subsequent medical intensive care measures which resulted in total recovery of a 42-year-old male. After admission to the medical intensive care unit the intoxicated patient was deeply hypotensive and needed fluid replacement, dobutamine and norepinephrine. Additionally insulin and calcium gluconate were given. Since hypotension persisted and the patient developed oliguria, terlipressin was applied and finally showed an effect on blood pressure and on urinary output. A volume overload of 7 L in the first 24 h resulted in a pulmonary edema. The patient was started on non-invasive ventilation with continuous positive airway pressure (CPAP) and frusemide was added to the therapy with good success. Quantitative determination of amlodipine in plasma samples was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The highest amlodipine concentrations was measured in the plasma sample collected approximately 8 h after ingestion of the drug, and was 393 microg/L. Four days later, it was possible to stop the treatment with catecholamines, at that time the amlodipine plasma concentration had declined to 132 microg/L, still tenfold higher than therapeutic (5-18 microg/L). Elimination half-life of amlodipine is approximately 55 h. After 6 days in the intensive care unit the patient was transferred to psychiatric treatment. Intensive care management and plasma levels in this intoxication case are compared to data from literature on other cases.     

Luminous lip-prints as criminal evidence

Luminescence is specially a useful property for the search of invisible evidences at the scene of a crime. In the latent fingerprints particular case, there are at one's disposal fluorescent reagents for their localization. The study of latent lip prints (that is lip prints from protective lipstick, or permanent or long-lasting lipstick that do not leave any visible marks) is more recent than fingerprints study. Because of the different composition of both types of prints, different reagents have been tried out on their developing. Although, lysochromes are particularly useful reagents to obtain latent lip prints, it may occur on coloured or multicoloured surfaces, the developing is not perceived due to contrast problems between the reagent and the surface where the print is searched. Again, luminescence offers the possibility to solve this problem. Nile Red is being studied as a potential developer for latent lip prints. The results on very old prints (over 1year) indicate that this reagent is highly efficient to get latent lip prints.     

Power rather than path dependency? The dynamics of institutional change under health care federalism

Proposals for government decentralization rank high on the political reform agenda of health systems worldwide. Their impact on welfare state performance and change, however, is still under theoretical scrutiny. This article examines the impact of devolution on the construction of the Spanish National Health Service (NHS) in an attempt to shed some light on this debate. Against widespread claims of path dependency, we argue that the specific nature of the devolution model developed in Spain, given the more egalitarian sociopolitical structure that resulted from democratization, fostered policy innovation and institutional change. Consolidation of an NHS system was compatible with some regional diversity and apparently prevented the rise of significant territorial inequalities. The Spanish case also suggests that policy change depends more on the distribution of social power than on institutions. It underlines the key role of financial and knowledge transfers vis-à-vis institutional reforms in effecting social change as well as the potential for state intervention in supporting the development of collective action resources by social groups.