The use of wastewater analysis in forensic intelligence: drug consumption comparison between Sydney and different European cities

© 2019, © 2019 The Author(s). Published by Taylor & Francis Group on behalf of the Academy of Forensic Science. Wastewater analysis offers objective and complementary information to illicit drug agencies by monitoring patterns of illicit drug consumption. In this study, wastewater samples from three different wastewater treatment plants in Sydney, Australia were collected in March 2016. Ten targeted drugs were analysed and temporal and geographical analyses were performed to obtain a better understanding of the type and amount of illicit drugs consumed in Sydney in comparison with similar studies conducted around Australia and in Europe. Among the targeted drugs, methamphetamine was consumed the most, followed by cocaine and 3,4-methylenedioxymethamphetamine (MDMA). Weekly patterns were observed where a peak during the weekend was present. The geographical analysis showed differences between the regions targeted. This observation may be related to socio-demographic aspects. The comparison of our study to other data sources from Australia showed a high consumption of methamphetamine in Sydney and Western Australia. The comparison between Sydney and different European cities revealed a difference in consumption, which is in line with traditional market indicators. The information obtained through wastewater analysis provides complementary information regarding illicit drug consumption, the size, and the evolution of the illicit drug market. This, ultimately, will assist authorities in making informed decisions

Single \pi^- production in np collisions for excess energies up to 90 MeV

The quasifree reaction np\to pp\pim was studied in a kinematically complete experiment by bombarding a liquid hydrogen target with a deuteron beam of momentum 1.85 GeV/c and analyzing the data along the lines of the spectator model. In addition to the three charged ejectiles the spectator proton was also detected in the large-acceptance time-of-flight spectrometer COSY-TOF. It was identified by its momentum and flight direction thus yielding access to the Fermi motion of the bound neutron and to the effective neutron 4-momentum vector $\mathbb{P}_n$ which differed from event to event. A range of almost 90 MeV excess energy above threshold was covered. Energy dependent angular distributions, invariant mass spectra as well as fully covered Dalitz plots were deduced. Sizeable $pp$ FSI effects were found as were contributions of $p$ and $d$ partial waves. The behavior of the elementary cross section $\sigma_{01}$ close to threshold is discussed in view of new cross section data. In comparison with existing literature data the results provide a sensitive test of the spectator model.Comment: 21 pages, 9 figures, 4 tables, submitted to EPJ

Observation of strong final-state effects in pi+ production in pp collisions at 400 MeV

Differential cross sections of the reactions $pp \to d\pi^+$ and $pp \to pn\pi^+$ have been measured at $T_p = 400$ MeV by detecting the charged ejectiles in the angular range $4^0 \leq \Theta_{Lab} \leq 21^\circ$. The deduced total cross sections agree well with those published previously for neighbouring energies. The invariant mass spectra are observed to be strongly affected by $\Delta$ production and $NN$ final-state interaction. The data are well described by Monte Carlo simulations including both these effects. The ratio of $pp \to pn\pi^+$ and $pp \to d\pi^+$ cross sections also compares favourably to a recent theoretical prediction which suggests a dominance of $np$-production in the relative $^3S_1$-state.Comment: 17 pages, 5 figure

Controlled assembly of SNAP-PNA-fluorophore systems on DNA templates to produce fluorescence resonance energy transfer

The SNAP protein is a widely used self-labeling tag that can be used for tracking protein localization and trafficking in living systems. A model system providing controlled alignment of SNAP-tag units can provide a new way to study clustering of fusion proteins. In this work, fluorescent SNAP-PNA conjugates were controllably assembled on DNA frameworks forming dimers, trimers, and tetramers. Modification of peptide nucleic acid (PNA) with the O6-benzyl guanine (BG) group allowed the generation of site-selective covalent links between PNA and the SNAP protein. The modified BG-PNAs were labeled with fluorescent Atto dyes and subsequently chemo-selectively conjugated to SNAP protein. Efficient assembly into dimer and oligomer forms was verified via size exclusion chromatography (SEC), electrophoresis (SDS-PAGE), and fluorescence spectroscopy. DNA directed assembly of homo- and hetero-dimers of SNAP-PNA constructs induced homo- and hetero-FRET, respectively. Longer DNA scaffolds controllably aligned similar fluorescent SNAP-PNA constructs into higher oligomers exhibiting homo-FRET. The combined SEC and homo-FRET studies indicated the 1:1 and saturated assemblies of SNAP-PNA-fluorophore:DNA formed preferentially in this system. This suggested a kinetic/stoichiometric model of assembly rather than binomially distributed products. These BG-PNA-fluorophore building blocks allow facile introduction of fluorophores and/or assembly directing moieties onto any protein containing SNAP. Template directed assembly of PNA modified SNAP proteins may be used to investigate clustering behavior both with and without fluorescent labels which may find use in the study of assembly processes in cells

Mitochondrial Diabetes in Children: Seek and You Will Find It

Maternally Inherited Diabetes and Deafness (MIDD) is a rare form of diabetes due to defects in mitochondrial DNA (mtDNA). 3243 A>G is the mutation most frequently associated with this condition, but other mtDNA variants have been linked with a diabetic phenotype suggestive of MIDD. From 1989 to 2009, we clinically diagnosed mitochondrial diabetes in 11 diabetic children. Diagnosis was based on the presence of one or more of the following criteria: 1) maculopathy; 2) hearing impairment; 3) maternal heritability of diabetes/impaired fasting glucose and/or hearing impairment and/or maculopathy in three consecutive generations (or in two generations if 2 or 3 members of a family were affected). We sequenced the mtDNA in the 11 probands, in their mothers and in 80 controls. We identified 33 diabetes-suspected mutations, 1/33 was 3243A>G. Most patients (91%) and their mothers had mutations in complex I and/or IV of the respiratory chain. We measured the activity of these two enzymes and found that they were less active in mutated patients and their mothers than in the healthy control pool. The prevalence of hearing loss (36% vs 75–98%) and macular dystrophy (54% vs 86%) was lower in our mitochondrial diabetic adolescents than reported in adults. Moreover, we found a hitherto unknown association between mitochondrial diabetes and celiac disease. In conclusion, mitochondrial diabetes should be considered a complex syndrome with several phenotypic variants. Moreover, deafness is not an essential component of the disease in children. The whole mtDNA should be screened because the 3243A>G variant is not as frequent in children as in adults. In fact, 91% of our patients were mutated in the complex I and/or IV genes. The enzymatic assay may be a useful tool with which to confirm the pathogenic significance of detected variants

Pregabalin in fibromyalgia - responder analysis from individual patient data

<p>Abstract</p> <p>Background</p> <p>Population mean changes are difficult to use in clinical practice. Responder analysis may be better, but needs validating for level of response and treatment duration. A consensus group has defined what constitutes minimal, moderate, and substantial benefit based on pain intensity and Patient Global Impression of Change scores.</p> <p>Methods</p> <p>We obtained individual patient data from four randomised double blind trials of pregabalin in fibromyalgia lasting eight to 14 weeks. We calculated response for all efficacy outcomes using any improvement (≥ 0%), minimal improvement (≥ 15%), moderate improvement (≥ 30%), substantial improvement (≥ 50%), and extensive improvement (≥ 70%), with numbers needed to treat (NNT) for pregabalin 300 mg, 450 mg, and 600 mg daily compared with placebo.</p> <p>Results</p> <p>Information from 2,757 patients was available. Pain intensity and sleep interference showed reductions with increasing level of response, a significant difference between pregabalin and placebo, and a trend towards lower (better) NNTs at higher doses. Maximum response rates occurred at 4-6 weeks for higher levels of response, and were constant thereafter. NNTs (with 95% confidence intervals) for ≥ 50% improvement in pain intensity compared with placebo after 12 weeks were 22 (11 to 870) for pregabalin 300 mg, 16 (9.3 to 59) for pregabalin 450 mg, and 13 (8.1 to 31) for pregabalin 600 mg daily. NNTs for ≥ 50% improvement in sleep interference compared with placebo after 12 weeks were 13 (8.2 to 30) for pregabalin 300 mg, 8.4 (6.0 to 14) for pregabalin 450 mg, and 8.4 (6.1 to 14) for pregabalin 600 mg. Other outcomes had fewer respondents at higher response levels, but generally did not discriminate between pregabalin and placebo, or show any dose response. Shorter duration and use of 'any improvement' over-estimated treatment effect compared with longer duration and higher levels of response.</p> <p>Conclusions</p> <p>Responder analysis is useful in fibromyalgia, particularly for pain and sleep outcomes. Some fibromyalgia patients treated with pregabalin experience a moderate or substantial pain response that is consistent over time. Short trials using 'any improvement' as an outcome overestimate treatment effects.</p

Pathogenic p62/SQSTM1 mutations impair energy metabolism through limitation of mitochondrial substrates

Abnormal mitochondrial function has been found in patients with frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). Mutations in the p62 gene (also known as SQSTM1) which encodes the p62 protein have been reported in both disorders supporting the idea of an ALS/FTD continuum. In this work the role of p62 in energy metabolism was studied in fibroblasts from FTD patients carrying two independent pathogenic mutations in the p62 gene, and in a p62-knock-down (p62 KD) human dopaminergic neuroblastoma cell line (SH-SY5Y). We found that p62 deficiency is associated with inhibited complex I mitochondrial respiration due to lack of NADH for the electron transport chain. This deficiency was also associated with increased levels of NADPH reflecting a higher activation of pentose phosphate pathway as this is accompanied with higher cytosolic reduced glutathione (GSH) levels. Complex I inhibition resulted in lower mitochondrial membrane potential and higher cytosolic ROS production. Pharmacological activation of transcription factor Nrf2 increased mitochondrial NADH levels and restored mitochondrial membrane potential in p62-deficient cells. Our results suggest that the phenotype is caused by a loss-of-function effect, because similar alterations were found both in the mutant fibroblasts and the p62 KD model. These findings highlight the implication of energy metabolism in pathophysiological events associated with p62 deficiency