Quantitative profiling of the human substantia nigra proteome from laser-capture microdissected FFPE tissue

Laser-capture microdissection (LCM) allows the visualization and isolation of morphologically distinct subpopulations of cells from heterogeneous tissue specimens. In combination with formalin-fixed and paraffin-embedded (FFPE) tissue it provides a powerful tool for retrospective and clinically relevant studies of tissue proteins in a healthy and diseased context. We first optimized the protocol for efficient LCM analysis of FFPE tissue specimens. The use of SDS containing extraction buffer in combination with the single-pot solid-phase-enhanced sample preparation (SP3) digest method gave the best results regarding protein yield and protein/peptide identifications. Microdissected FFPE human substantia nigra tissue samples (~3,000 cells) were then analyzed, using tandem mass tag (TMT) labeling and LC-MS/MS, resulting in the quantification of &gt;5,600 protein groups. Nigral proteins were classified and analyzed by abundance, showing an enrichment of extracellular exosome and neuron-specific gene ontology (GO) terms among the higher abundance proteins. Comparison of microdissected samples with intact tissue sections, using a label-free shotgun approach, revealed an enrichment of neuronal cell type markers, such as tyrosine hydroxylase and alpha-synuclein, as well as proteins annotated with neuron-specific GO terms. Overall, this study provides a detailed protocol for laser-capture proteomics using FFPE tissue and demonstrates the efficiency of LCM analysis of distinct cell subpopulations for proteomic analysis using low sample amounts.</p

‘What’s it like to have ME?’ The discursive construction of ME in computer-mediated communication and face-to-face interaction

ME/CFS (chronic fatigue syndrome) is a debilitating illness for which no cause or medical tests have been identified. Debates over its nature have generated interest from qualitative researchers. However, participants are difficult to recruit because of the nature of their condition. Therefore, this study explores the utility of the internet as a means of eliciting accounts. We analyse data from focus groups and the internet in order to ascertain the extent to which previous research findings apply to the internet domain. Interviews were conducted among 49 members of internet (38 chatline, 11 personal) and 7 members of two face-to-face support groups. Discourse analysis of descriptions and accounts of ME/CFS revealed similar devices and interactional concerns in both internet and face-to-face communication. Participants constructed their condition as serious, enigmatic and not psychological. These functioned to deflect problematic assumptions about ME/CFS and to manage their accountability for the illness and its effects

Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster

Spinal Muscular Atrophy (SMA) is caused by homozygous mutations in the human survival motor neuron 1 (SMN1) gene. SMN protein has a well-characterized role in the biogenesis of small nuclear ribonucleoproteins (snRNPs), core components of the spliceosome. SMN is part of an oligomeric complex with core binding partners, collectively called Gemins. Biochemical and cell biological studies demonstrate that certain Gemins are required for proper snRNP assembly and transport. However, the precise functions of most Gemins are unknown. To gain a deeper understanding of the SMN complex in the context of metazoan evolution, we investigated its composition in Drosophila melanogaster. Using transgenic flies that exclusively express Flag-tagged SMN from its native promoter, we previously found that Gemin2, Gemin3, Gemin5, and all nine classical Sm proteins, including Lsm10 and Lsm11, co-purify with SMN. Here, we show that CG2941 is also highly enriched in the pulldown. Reciprocal co-immunoprecipitation reveals that epitope-tagged CG2941 interacts with endogenous SMN in Schneider2 cells. Bioinformatic comparisons show that CG2941 shares sequence and structural similarity with metazoan Gemin4. Additional analysis shows that three other genes (CG14164, CG31950 and CG2371) are not orthologous to Gemins 6-7-8, respectively, as previously suggested. In D.melanogaster, CG2941 is located within an evolutionarily recent genomic triplication with two other nearly identical paralogous genes (CG32783 and CG32786). RNAi-mediated knockdown of CG2941 and its two close paralogs reveals that Gemin4 is essential for organismal viability

Does child abuse predict a population segment with large economic burden?

Objectives: The enormous societal and individual consequences of mental health disorders and detrimental health behaviours in the general population are of paramount concern. Many argue that ‘prevention is the best cure’, pushing for the implementation of early (preventive) interventions. Key questions regarding early interventions include which population segment to target for screenings and what information these screenings should focus on. In line with previous efforts, this study aimed to identify which population segment holds the majority (≥ 80 %) of different economically costly outcomes in society, and whether child abuse before the age of 16 years predicts being part of that population segment. Study design: Epidemiological cohort study. Methods: This study used the Netherlands Mental Health Survey and Incidence Study-2, a Dutch epidemiological cohort study including 6646 adults aged 18–64 years at baseline, spanning four timepoints from 2007 to 2018. Cumulative distributions were computed to identify high-cost population segments of economically costly outcomes in adulthood (i.e., mental and physical health [behaviours], unemployment and work absenteeism). Child abuse was examined as a potential predictor of these segments and the risk of multiple high-cost population segment membership was investigated by conducting Poisson regressions. Results: A 20 % population segment carried between 42 % and 100 % of economically costly outcomes. Being exposed to more child abuse predicted being in a high-cost population segment, albeit with small effect sizes. Being exposed to more child abuse also predicted belonging to multiple high-cost population segments across different economically costly outcomes. Conclusions: The study findings have implications for policy makers. Emphasis should be placed on prevention aimed at identifying potential members of multiple high-cost population segments.</p

Insomnia and the incidence, recurrence and persistence of common mental disorders:Sex-differences in the general population

Insomnia is common throughout the population and thought to be a risk factor for mental disorders. We assessed the association of insomnia symptoms with incidence, recurrence and persistence of mood, anxiety and substance use disorders. In 4007 participants (55 % women, mean age 51.0 ± 12.3) of the population-based Netherlands Mental Health Survey and Incidence Study (NEMESIS), having insomnia symptoms increased the odds of developing, recurring and persisting mood disorders, mostly in men. Insomnia only associated with recurring anxiety disorders, particularly in women, and not with substance use disorders. Treating insomnia may aid recovery and prevention of mental disorders, particularly mood disorders.</p

Enhanced activity of desilicated Cu-SSZ-13 for the selective catalytic reduction of NOx and its comparison with steamed Cu-SSZ-13

Mesoporous Cu-SSZ-13 was created by first synthesizing zeolite H-SSZ-13 and subsequently desilicating the material by base leaching using NaOH in different concentrations. The catalyst materials were prepared by ion exchanging the leached samples back to their acidic form using NH4NO3, and to their active Cu form by ion exchanging them with CuSO4. For comparison, H- and Cu-SSZ-13 were steamed using a wide variety of different conditions. Using a 0.10 M NaOH solution for base leaching, it was found that Cu-SSZ-13 becomes more active in the selective catalytic reduction of NOx with NH3 (NH3-SCR) over the entire temperature region but especially in the low temperature region (<200 °C). This increase could be explained by a decrease in pore diffusion limitations due to the introduction of mesopores on the outside of the zeolite crystals but keeping the chemical environment of the catalyst nearly the same as that of the parent material. Higher base leaching concentrations do, however, lead to a decrease in the amount of Brønsted acid sites, pore volume and accessible surface area, accompanied by a decrease in NH3-SCR activity. Ar physisorption coupled with SEM and confocal fluorescence microscopy in combination with two differently sized fluorescent organic probe molecules (i.e., 4-(4-dimethyl-aminostyryl)-1-methyl-pyridinium-iodide and 4-(4-dicyclohexyl-aminostyryl)-1-methyl-pyridinium-iodide) show an increase in the external surface area due to the creation of mesopores. The development of mesoporosity starts from the crystal surface and continues into the crystal with increasing alkaline solution strength, but under our conditions it never reaches the center. On the other hand, zeolite steaming did not successfully introduce mesoporosity and mainly managed to deactivate the Cu-SSZ-13 zeolite catalysts

The global governance of human cloning: the case of UNESCO

Since Dolly the Sheep was cloned in 1996, the question of whether human reproductive cloning should be banned or pursued has been the subject of international debate. Feelings run strong on both sides. In 2005, the United Nations adopted its Declaration on Human Cloning to try to deal with the issue. The declaration is ambiguously worded, prohibiting “all forms of human cloning inasmuch as they are incompatible with human dignity and the protection of human life”. It received only ambivalent support from UN member states. Given this unsatisfactory outcome, in 2008 UNESCO (the United Nations Educational, Scientific and Cultural Organization) set up a Working Group to investigate the possibility of a legally binding convention to ban human reproductive cloning. The Working Group was made up of members of the International Bioethics Committee, established in 1993 as part of UNESCO’s Bioethics Programme. It found that the lack of clarity in international law is unhelpful for those states yet to formulate national regulations or policies on human cloning. Despite this, member states of UNESCO resisted the idea of a convention for several years. This changed in 2015, but there has been no practical progress on the issue. Drawing on official records and first-hand observations at bioethics meetings, this article examines the human cloning debate at UNESCO from 2008 onwards, thus building on and advancing current scholarship by applying recent ideas on global governance to an empirical case. It concludes that, although human reproductive cloning is a challenging subject, establishing a robust global governance framework in this area may be possible via an alternative deliberative format, based on knowledge sharing and feasibility testing rather than the interest-based bargaining that is common to intergovernmental organizations and involving a wide range of stakeholders. This article is published as part of a collection on global governance

Validity of the five-item mental health inventory for screening current mood and anxiety disorders in the general population

Objectives: The Mental Health Inventory (MHI-5) is frequently used as a screener for mood and anxiety disorders. However, few population-based studies have validated it against a diagnostic instrument assessing disorders following current diagnostic criteria. Methods: Within the third Netherlands Mental Health Survey and Incidence Study (NEMESIS-3), a representative population-based study of adults (N = 6194; age: 18–75 years), the MHI-5 was used to measure general mental ill-health in the past month. Presence of mood (major depressive disorder, persistent depressive disorder, or bipolar disorder) and anxiety disorders (panic disorder, agoraphobia, social phobia, or generalized anxiety disorder) in the past month was assessed with a slightly modified version of the Composite International Diagnostic Interview 3.0 per the Diagnostic and Statistical Manual of Mental disorders-5. Results: The MHI-5 was good to excellent at distinguishing people with and without a mood disorder, an anxiety disorder, and any mood or anxiety disorder. The cut-off value associated with the highest sensitivity and highest specificity for mood disorder was ≤68, and ≤76 for an anxiety disorder or any mood or anxiety disorder. Conclusions: The MHI-5 can identify individuals at high risk of a current mood or anxiety disorder in the general population when diagnostic interviews are too time consuming.</p

The design of a randomised controlled trial to evaluate the (cost-) effectiveness of the posterolateral versus the direct anterior approach for THA (POLADA - trial)

Background: Total hip arthroplasty (THA) is one of the most successful orthopaedic procedures. Because of the increasing number of THAs, a growing demand for faster recovery and a greater emphasis on cost-effectiveness, minimally invasive THAs have been introduced in the last decades. The direct anterior approach is a minimally invasive, tissue-sparing approach in which intermuscular planes are used. Theoretically, this approach should result in a faster recovery of physical functioning and higher health-related quality of life. Methods/design: A randomised controlled trial will be performed. Patients will be randomly allocated to undergo THA by means of the anterior or posterolateral approach. Both the intervention and control group will consist of two subgroups: 1) patients with a good bone stock who will receive an uncemented femoral stem, and 2) patients with a poor bone stock who will receive a cemented femoral stem. Patients between 18 and 90 years with primary or secondary osteoarthritis will be included. Physical functioning and health-related quality of life will be assessed by means of questionnaires. Additionally, performance based tests will be performed to objectively assess the physical functioning. Cost-effectiveness will be assessed by obtaining data on medical costs in and outside the hospital and other nonmedical costs. Measurements will take place preoperatively, two and six weeks, three months and one year postoperatively. Discussion: There is some evidence that the anterior approach results in reduced tissue damage and faster recovery in the direct postoperative period, compared to the posterolateral approach. However, there is still a lack of well-designed studies that have confirmed the better outcomes and cost-effectiveness of the anterior approach. Therefore, the purpose of this study is to assess the physical functioning, health related quality of life and the cost-effectiveness of the anterior approach, compared to the conventional posterolateral approach