RNA/DNA co-analysis from blood stains--results of a second collaborative EDNAP exercise

A second collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling was organized by the European DNA Profiling Group (EDNAP). Six human blood stains, two blood dilution series (5-0.001 μl blood) and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by the participating laboratories using a RNA/DNA co-extraction or solely RNA extraction method. Two novel mRNA multiplexes were used for the identification of blood: a highly sensitive duplex (HBA, HBB) and a moderately sensitive pentaplex (ALAS2, CD3G, ANK1, SPTB and PBGD). The laboratories used different chemistries and instrumentation. All of the 18 participating laboratories were able to successfully isolate and detect mRNA in dried blood stains. Thirteen laboratories simultaneously extracted RNA and DNA from individual stains and were able to utilize mRNA profiling to confirm the presence of blood and to obtain autosomal STR profiles from the blood stain donors. The positive identification of blood and good quality DNA profiles were also obtained from old and compromised casework samples. The method proved to be reproducible and sensitive using different analysis strategies. The results of this collaborative exercise involving a RNA/DNA co-extraction strategy support the potential use of an mRNA based system for the identification of blood in forensic casework that is compatible with current DNA analysis methodology.     

Tissue selectivity of endothelin

The effects of endothelin, a potent endogenous vasoconstrictor peptide, were examined in a range of vascular and non-vascular tissues. At concentrations that cause vasoconstriction in portal vein and aorta, the peptide strongly contracted rat uterus, trachea and vas deferens, but not guinea pig ileum. Nifedipine, a dihydropyridine calcium anatgonist, partially inhibited these contractions. Endothelin had no inotropic or chronotropic effect on the isolated rat heart. The peptide did not modulate secretion at the neuromuscular junction, from adrenal medullary cells or neutrophils, nor affect secretion or aggregation of platelets. The tissue responsiveness to endothelin was not the same as the tissue distribution of dihydropyridine receptors. This supports the idea that endothelin interacts with a specific receptor distinct from dihydropyridine sensitive calcium channels. The contractile effect of endothelin on non vascular smooth muscle suggests that the concept of endothelium dependent modulation of vascular smooth muscle tone may be extended to include epithelium dependent modulation of non vascular tissues.     

Dose-related effects of perfluorodecanoic acid on growth,feed intake and hepatic peroxisomal β-oxidation

The effects of the persistent peroxisome proliferator, perfluorodecanoic acid (PFDA), on growth, feed intake and the enzyme activities associated with peroxisomal β-oxidation were studied in female Sprague Dawley rats. Rats received one of six levels of PFDA (0, 0.3, 1.0, 3.0, 10.0 or 30.0 mg/kg/injection) in four IP doses at 2-week intervals. Rats with cumulative doses of ≤ 12.0 mg/kg did not differ from control rats in growth or feed intake, while rats receiving cumulative doses of ≥40 mg/kg lost weight and decreased their feed intake. Rats which received cumulative doses between these levels increased their feed intake but did not significantly alter their body weight. Total peroxisomal β-oxidation was decreased in a dose-related manner, whereas the liver to body weight ratio and the activities of individual enzymes comprising the peroxisomal β-oxidation system, namely fatty acyl-CoA oxidase, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and thiolase, were increased. This study clearly shows that the inhibition of peroxisomal ß-oxidation by PFDA is not reflected in the in vitro measurement of the individual enzyme activities comprising this pathway.     

The recognition site for hepatic clearance of plasma kallikrein is on its heavy chain and is latent on prokallikrein.

We partially purified the glycoproteins prokallikrein and kallikrein from rat plasma. The purification of rat plasma kallikrein may result in two forms: an intact form (alpha, M(r) 84-87 kDa) and a partially degraded form (beta, M(r) 46-51 kDa). The alpha-form is composed of a heavy chain (M(r) 50 kDa) and a light chain (M(r) 34-37 kDa) linked by a disulfide bond. The catalytic site is found on the light chain. The beta-form has a partially degraded heavy chain (M(r) 28 kDa). Using a preparation of exsanguinated and perfused rat liver, we verified that rat plasma prokallikrein is not activated by the liver and that neither the proenzyme nor the light chain is removed by the organ. Both forms (alpha and beta) of the active enzyme are similarly removed from the perfusate. We also observed that the clearance of plasma kallikrein is temperature-dependent, and not affected by substances that inhibit binding to galactosyl-, mannosyl-, fucosyl- or phosphomannosyl-specific lectins, but inhibited by beta-galactosides. We suggest that: (a) the binding site to hepatocytes is latent on prokallikrein and is located on its heavy chain, more specifically on the 28-kDa fragment still present in the beta form of the active enzyme and (b) plasma kallikrein is recognized by an S-type lectin.     

Comorbidity of substance use disorders with mood and anxiety disorders: Results of the international consortium in psychiatric epidemiology

This article reports the results of a cross-national investigation of patterns of comorbidity between substance use and psychiatric disorders in six studies participating in the International Consortium in Psychiatric Epidemiology. In general, there was a strong association between mood and anxiety disorders as well as conduct and antisocial personality disorder with substance disorders at all sites. The results also suggest that there is a continuum in the magnitude of comorbidity as a function of the spectrum of substance use category (use, problems, dependence), as well as a direct relationship between the number of comorbid disorders and increasing levels of severity of substance use disorders (which was particularly pronounced for drugs). Finally, whereas there was no specific temporal pattern of onset for mood disorders in relation to substance disorders, the onset of anxiety disorders was more likely to precede that of substance disorders in all countries. These results illustrate the contribution of cross-national data to understanding the patterns and risk factors for psychopathology and substance use disorders.     

Infection by cytomegalovirus in patients with acquired immunodeficiency syndrome (AIDS): clinical, virological, and histopathological correlations]

Between April 1986 and June 1987, 50 patients meeting the CDC criteria for AIDS were studied for serological and virological evidence of CMV infection. Attempts for virus isolation from peripheral blood, urine and saliva were performed in cell culture lines of human foreskin fibroblasts and CMV specific IgG and IgM were assayed by IFI and IgG by ELISA. A total of 121 blood, 119 urine and 96 saliva samples were collected. During the study period viremia was noted at least once in 12.5%, viruria in 23.2%, and excretion in saliva in 21.9%. When admitted in the study, 20% (10/50) of the patients had anti-CMV IgM antibodies and 100% (50/50) of them had IgG anti-CMV antibodies (IFI). Five of the 40 patients IgM negative at admission presented anti-CMV IgM antibodies during the study, suggesting CMV reactivation or reinfection. Active CMV infection based on virus isolation and/or IgM positivity was demonstrated in 60% of the patients. Histopathological studies were performed in 24 patients. CMV was found in 50% of the autopsies, mainly in the digestive system, lungs and adrenals. There was no correlation between clinical, virological (serology and isolation) and histopathological diagnosis.     

Topography of choline acetyltransferase Immunoreactive Neurons and fibers in the rat spinal cord

The topography of choline acetyltransferase immunoreactivity was studied in the rat spinal cord with a monoclonal antibody. Cholinergic fibers were most prominent in lamina III of the dorsal horn and originated from cholinergic neurons within the spinal cord. Lamina X, which was rich in cholinergic neurons and fibers, provided cholinergic interconnections between the dorsal, intermediate and ventral gray. Within the ventral gray, choline acetyltransferase immunoreactive boutons were found on motor neurons. This study suggests that the cholinergic innervation of the spinal cord arises from neurons intrinsic to the spinal cord. The cholinergic neurons within the spinal cord may provide several, overlapping levels of regulation of spinal cord neurons.