Optimization of a duplex amplification and sequencing strategy for the HVI/HVII regions of human mitochondrial DNA for forensic casework

A duplex primer set for the amplification of mitochondrial DNA HVI and HVII control regions was evaluated for the optimization of a DNA sequencing protocol suitable for forensic casework. HVI and HVII products, with the absence of non-specific products, could be detected by agarose gel electrophoresis when as little as 0.5 and 0.1pg of DNA were amplified for 34 and 38 cycles, respectively. Because HVI and HVII amplicons are co-synthesized in the duplex PCR, fewer steps are required (lessening the risk of cross contamination events) and more frugal use of precious extracted DNA samples is possible, both desirable features for forensic casework. The ABI Prism BigDyetrade mark version 1.1 chemistry provided high quality sequencing data, with little or no background noise and uniform peak heights, outcomes that favored reliable detection of heteroplasmy, particularly at early sequence reads (<40 bases). Optimal compromise between sensitivity and sequence accuracy in the absence of noise was achieved starting at 150 mitochondrial genome copies. The protocol is effective (no sequence errors) with highly degraded DNA (average detectable template size of 200bp). Dual artificial template mixtures with the minor component at 15% suggests that heteroplasmy should be detected at this level with confidence.     

Inmate family functioning

Two-hundred-nine noninmates and 169 inmates completed questionnaires that assessed retrospective perceptions of 12 dimensions of family life and one overall assessment of quality of family life. Between the inmates and noninmates, means for all dependent variables differed significantly except for self-reliance; however, meaningful eta-squares were found only for dimensions of bridging, disengagement, and quality of life. Among the independent-samples t tests for gender, eta-squares were not meaningful. Implications for family life interventions in correctional facilities are suggested.     

Mass spectral and NMR spectral data of two new designer drugs with an alpha-aminophenone structure: 4'-methyl-alpha-pyrrolidinohexanophenone and 4'-methyl-alpha-pyrrolidinobutyrophenone

This study presents and discusses the nuclear magnetic resonance (NMR) spectroscopic and mass spectroscopic data of the new designer drug 4'-methyl-alpha-pyrrolidinobutyrophenone (MPBP) and its homolog 4'-methyl-alpha-pyrrolidinohexanophenone (MPHP) which were seized in 2004 and 2000 in Germany for the first time. The structure elucidation of the aliphatic part of MPBP was carried out by product ion spectroscopy of the immonium ion formed after electron ionization as well as with 1H and 13C NMR. Product ion spectroscopy of immonium ions again proved to be a powerful tool to determine the structure of designer drugs and to distinguish between isobaric structures of the alkyl-amino moiety.     

Effect of ethanol on cardiac single sodium channel gating

Alcohol in modest and higher doses has the potential to induce cardiac arrhythmias. The most famous alcohol-related arrhythmia is the "holiday heart syndrome". Furthermore, there is a clear association between excessive alcohol consumption and the risk of sudden cardiac death. However, the acute effects of ethanol on arrhythmia induction are not well understood. The effect of ethanol on single cardiac sodium channels has not been studied yet. To elucidate the effect of ethanol on human cardiac sodium channels we performed a patch clamp study in HEK-293 cells overexpressing the human cardiac sodium channel. We used HEK-293 cells overexpressing the human cardiac sodium channel (Na(1.5)). Single channel gating was investigated by the cell-attached patch clamp technique. Sodium channel currents were elicited by depolarizing pulses from -120 to -20mV for a duration of 150ms. Single channel availability, open probability and peak average current were assessed baseline and after addition of ethanol in increasing concentrations (0.50 per thousand (10.9mM), 1.00 per thousand (21.7mM), 2.00 per thousand (43.5mM) and 4.00 per thousand (87.0mM)). We found a concentration-dependent reduction of open probability which was statistically significant at 2.00 per thousand ethanol (66.5+/-14% of control). At higher concentrations (4.00 per thousand) also availability decreased to 66.5+/-11.0% of control. This resulted in a significant decrease of peak average current at 2.00 per thousand and at 4.00 per thousand ethanol (61.8+/-7.4 and 53.0+/-8.2% of control). For the first time the present study demonstrates acute inhibitory effects of ethanol on single cardiac sodium channel gating and provides one potential mechanism for the well known clinical observation that ethanol triggers supraventricular and ventricular arrhythmias.     

Quasi-experimental designs for community-level public health violence reduction interventions: a case study in the challenges of selecting the counterfactual

Objectives

We highlight the importance of documenting the step-by-step processes used for the selection of comparison areas when evaluating a community-level intervention that targets a large-scale community.

Methods

We demonstrate the proposed method using a propensity score matching framework for an impact analysis of the Cure Violence Public Health Model in Philadelphia. To select comparison communities, propensity score models are run using different levels of aggregation to define the intervention site. We discuss the trade-offs made.

Results

We find wide variation in documentation and explanation in the extant literature of the methods used to select comparison communities. The size of the unit of analysis at which a community is measured complicates the decision processes, and in turn, can affect the validity of the counterfactual.

Conclusions

It is important to carefully consider the unit of analysis for measurement of comparison communities. Assessing the geographic clustering of matched communities to mirror that of the treated community holds conceptual appeal and represents a strategy to consider when evaluating community-level interventions taking place at a large scale. Regardless of the final decisions made in the selection of the counterfactual, the field could benefit from more systematic diagnostic tools that document and guide the steps and decisions along the way, and ask: “could there have been another way of doing each step, and what difference would this have made?” Overall, across community-level evaluations that utilize quasi-experimental designs, documentation of the counterfactual selection process will provide a more fine-grained understanding of causal inference.