Placental DAPK1 and autophagy marker LC3B-II are dysregulated by TNF-α in a gestational age-dependent manner

Autophagy, a cell-survival process responsible for degradation of protein aggregates and damaged organelles, is increasingly recognized as another mechanism essential for human placentation. A substantial body of experiments suggests inflammation and oxidative stress as the underlying stimuli for altered placental autophagy, giving rise to placenta dysfunction and pregnancy pathologies. Here, the hypothesis is tested whether or not pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-{alpha} are able to influence the expression profile of autophagy genes in human first-trimester villous placenta. Autophagy-focused qPCR arrays identified substantial downregulation of death-associated protein kinase 1 (DAPK1) in first-trimester placental explants in response to IL-6 and TNF-{alpha}, respectively. Immunohistochemistry of placental explants detected considerable DAPK1 staining in placental macrophages, villous cytotrophoblasts and less intense in the syncytiotrophoblast. Both immunohistochemistry and Western blot showed decreased DAPK1 protein in TNF-{alpha}-treated placental explants compared to control. On cellular level, DAPK1 expression decreased in SGHPL-4 trophoblasts in response to TNF-{alpha}. Observed changes in the expression profile of autophagy-related genes were reflected by significantly decreased lipidation of autophagy marker microtubule-associated protein light chain 3 beta (LC3B-II) in first trimester placental explants in response to TNF-{alpha}. Analysis of TNF-{alpha}-treated term placental explants showed decreased DAPK1 protein, whereas in contrast to first-trimester LC3B expression and lipidation increased. Immunohistochemistry of placental tissues from early-onset preeclampsia (PE) showed less DAPK1 staining, when compared to controls. Accordingly, DAPK1 mRNA and protein were decreased in primary trophoblasts isolated from early-onset PE, while LC3B-I and -II were increased. Results from this study suggest that DAPK1, a regulator of apoptosis, autophagy and programmed necrosis, decreases in human placenta in response to elevated maternal TNF-{alpha}, irrespective of gestational age. In contrast, TNF-{alpha} differentially regulates levels of autophagy marker LC3B in human placenta over gestation

Using Bayesian networks to guide the assessment of new evidence in an appeal case.

When new forensic evidence becomes available after a conviction there is no systematic framework to help lawyers to determine whether it raises sufficient questions about the verdict in order to launch an appeal. This paper presents such a framework driven by a recent case, in which a defendant was convicted primarily on the basis of audio evidence, but where subsequent analysis of the evidence revealed additional sounds that were not considered during the trial. The framework is intended to overcome the gap between what is generally known from scientific analyses and what is hypothesized in a legal setting. It is based on Bayesian networks (BNs) which have the potential to be a structured and understandable way to evaluate the evidence in a specific case context. However, BN methods suffered a setback with regards to the use in court due to the confusing way they have been used in some legal cases in the past. To address this concern, we show the extent to which the reasoning and decisions within the particular case can be made explicit and transparent. The BN approach enables us to clearly define the relevant propositions and evidence, and uses sensitivity analysis to assess the impact of the evidence under different assumptions. The results show that such a framework is suitable to identify information that is currently missing, yet clearly crucial for a valid and complete reasoning process. Furthermore, a method is provided whereby BNs can serve as a guide to not only reason with incomplete evidence in forensic cases, but also identify very specific research questions that should be addressed to extend the evidence base and solve similar issues in the future.This research was funded by the Engineering and Physical Sciences Research Council of the UK through the Security Science Doctoral Research Training Centre (UCL SECReT) based at University College London (EP/G037264/1), and the European Research Council (ERC-2013-AdG339182-BAYES_KNOWLEDGE)

A comment on the PCAST report:skip the “match”/“non-match” stage

This letter comments on the report “Forensic science in criminal courts: Ensuring scientific validity of feature-comparison methods” recently released by the President's Council of Advisors on Science and Technology (PCAST). The report advocates a procedure for evaluation of forensic evidence that is a two-stage procedure in which the first stage is “match”/“non-match” and the second stage is empirical assessment of sensitivity (correct acceptance) and false alarm (false acceptance) rates. Almost always, quantitative data from feature-comparison methods are continuously-valued and have within-source variability. We explain why a two-stage procedure is not appropriate for this type of data, and recommend use of statistical procedures which are appropriate

Determination of time since deposition of bloodstains through NIR and UV–Vis spectroscopy – A critical comparison

Time elapsed since bloodstain deposition is a crucial aspect in forensic investigations, where non-destructive spectroscopic methods play a pivotal role. While extensive research has been conducted by UV–Vis spectroscopy, showcasing its utility in specific cases, there is still a paucity of studies based on NIR spectroscopy, which has the potential to overcome the limitations of the UV–Vis-based methods. To compensate for this disequilibrium, the present study aimed to evaluate the NIR applicability for estimating the age of forensic bloodstains and develop a performance comparison with UV–Vis spectroscopy methods. Capillary blood was sampled and subjected to a 16-day aging, during which it was repeatedly analyzed using both spectroscopic methods. Subsequently, chemometric analysis was applied to process the spectral data and independently assess the methods’ performance. Classical preprocessing transforms (i.e., Savitzky-Golay derivatives and SNV transform) were used together with more targeted strategies, such as class centering, whose benefit was highlighted by PCA. Lastly, PLS regression models were computed to evaluate the effectiveness of both spectroscopic methods in estimating the time elapsed since blood trace deposition. Comparable root mean square errors in prediction (RMSEP) – 40 and 55 h for UV–Vis and NIR spectroscopy, respectively – were observed for both techniques, featuring an improvement with respect to the existing literature for NIR spectroscopy. Data fusion strategies for a multi-instrumental platform were also explored, evaluating advantages and disadvantages of low-level and mid-level approaches. The results indicated that NIR spectroscopy integrated with adequate chemometric strategies deserves increased appreciation in forensic bloodstain dating

The effect of environmental conditions and deposition substrate on bloodstains ageing: a spectroscopic approach to target a forensic analytical problem

Determining time since deposition of bloodstains can be a potent tool for the forensic chemist in resolving legal cases. This topic has been investigated through several analytical techniques, mainly spectroscopies, such as ultraviolet-visible (UV-Vis), Raman, mid and near infrared (MIR and NIR). In this context, the contribution of chemometrics proved to be essential to gain understanding of the subtle spectral changes ongoing during the ageing of this complex biological fluid. Until now, reduced attention has been devoted to the contribution of environmental conditions and deposition substrate which may have a significant influence on speed and kinetics of blood degradation. Therefore, understanding which factors influence this process (and to what extent) is crucial to avoid over/under estimations of bloodstain age. To study in depth the impact of such factors, bloodstains were subjected to ageing under controlled temperature (T) and humidity (RH%) in a constant climate chamber (KMF115 – BINDER GmbH, Tuttlingen, Germany), mimicking two environmental conditions with a proper day-night alternance: condition CW – day: T = 25°C, RH% = 75%; night: T = 10°C, RH% = 90%; condition HD – day: T = 35°C, RH% = 60%; night: T = 20°C, RH% = 75%. The effect of direct light exposure was investigated as well, performing the ageing sessions either with or without a window that filters the light going from the lamp to the samples. Six 20-μl bloodstains (from two different donors) were deposited on each of four different substrates, namely hydrophilic fabric (cotton), hydrophobic fabric (polyblend), metal (knife) and glass, making a total of 96 samples. Their ageing was followed for 12 days with two different analytical techniques: a handheld NIR device (MicroNIR® – Viavi Solutions Inc., Santa Rosa, California, USA), working in the spectral range 900-1700 nm; a confocal Raman microscope (InViaTM – Renishaw plc, Wotton-under-Edge, Gloucestershire, UK), acquiring in the Raman shift range 150-1900 cm-1. The obtained spectra were pre-processed for minimizing the unwanted systematic effects and subjected to chemometric analysis. Exploratory data analysis by means of principal component analysis (PCA) showed that NIR spectra are more subjected to the effect of the substrate, when compared with Raman spectra. It was observed that absorbing substrates (cotton, polyblend) yield a more efficient time trend model when analysed with NIR spectroscopy, while non-absorbing substrates (glass, metal) gave more consistent results with Raman spectroscopy. PCA also evidenced shifts between sessions carried out at different temperature and humidity, suggesting that environmental conditions may determine not only a faster/slower ageing, but also a different ageing kinetics. Then, ANOVA simultaneous component analysis (ASCA) was applied to quantify the effect of each factor (temperature, humidity, substrate, light exposure) on the ageing of bloodstains, to understand which conditions are most important to be controlled in real case scenarios. The present study confirmed that determination of time since deposition of bloodstains is strongly related to environmental conditions and deposition substrate and, therefore, that tailoring models to take into account these factors is preferable

ENFSI guideline for evaluative reporting in forensic science

Criminal Justice: Legitimacy, accountability, and effectivit

Outcome of infantile nephropathic cystinosis depends on early intervention, not genotype: A multicenter sibling cohort study.

Infantile nephropathic cystinosis (INC) is an inheritable lysosomal storage disorder characterized by lysosomal cystine accumulation, progressive kidney disease, and multiple extrarenal complications (ERCs). Cysteamine postpones the onset of end-stage kidney disease (ESKD) and reduces the incidence of ERCs; however, cysteamine is generally initiated upon establishment of the renal Fanconi syndrome (FS) and partial loss of kidney function, whereas data on long-term effects of cysteamine administered from neonatal age are lacking. An international multicenter retrospective cohort study of siblings with INC was set up to investigate the outcome in relation to age at initiation of cysteamine versus CTNS genotype, with attention to patients treated with cysteamine from neonatal age. None of the siblings treated from neonatal age (n = 9; age 10 ± 6 years) had reached ESKD, while 22% of their index counterparts (n = 9; age 14 ± 5 years) had commenced renal replacement therapy. Siblings treated with cysteamine from the onset of symptoms at a younger age compared with their index counterparts, reached ESKD at a significant older age (13 ± 3 vs. 10 ± 3 years, p = 0.002). In contrast, no significant difference in ERCs was observed between sibling and index patients, independently from the age at initiation of cysteamine. The CTNS genotype had no impact on the overall outcome in this cohort. In INC, presymptomatic treatment with cysteamine results in a better renal outcome in comparison to treatment initiated from the onset of symptoms. This justifies including cystinosis into newborn screening programs. SYNOPSIS: In infantile nephropathic cystinosis, presymptomatic treatment with cysteamine improves the renal outcome which justifies the inclusion of cystinosis into newborn screening programs