Analysis of Global Sumoylation Changes Occurring during Keratinocyte Differentiation

Sumoylation is a highly dynamic process that plays a role in a multitude of processes ranging from cell cycle progression to mRNA processing and cancer. A previous study from our lab demonstrated that SUMO plays an important role in keratinocyte differentiation. Here we present a new method of tracking the sumoylation state of proteins by creating a stably transfected HaCaT keratinocyte cell line expressing an inducible SNAP-SUMO3 protein. The SNAP-tag allows covalent fluorescent labeling that is denaturation resistant. When combined with two-dimensional gel electrophoresis, the SNAP-tag technology provides direct visualization of sumoylated targets and can be used to follow temporal changes in the global cohort of sumoylated proteins during dynamic processes such as differentiation. HaCaT keratinocyte cells expressing SNAP-SUMO3 displayed normal morphological and biochemical features that are consistent with typical keratinocyte differentiation. SNAP-SUMO3 also localized normally in these cells with a predominantly nuclear signal and some minor cytoplasmic staining, consistent with previous reports for untagged SUMO2/3. During keratinocyte differentiation the total number of proteins modified by SNAP-SUMO3 was highest in basal cells, decreased abruptly after induction of differentiation, and slowly rebounded beginning between 48 and 72 hours as differentiation progressed. However, within this overall trend the pattern of change for individual sumoylated proteins was highly variable with both increases and decreases in amount over time. From these results we conclude that sumoylation of proteins during keratinocyte differentiation is a complex process which likely reflects and contributes to the biochemical changes that drive differentiation

Good on paper: the gap between programme theory and real-world context in Pakistan's Community Midwife programme

Objective To understand why skilled birth attendance—an acknowledged strategy for reducing maternal deaths—has been effective in some settings but is failing in Pakistan and to demonstrate the value of a theory-driven approach to evaluating implementation of maternal healthcare interventions. Design Implementation research was conducted using an institutional ethnographic approach. Setting and population National programme and local community levels in Pakistan. Methods Observations, focus group discussions, and in-depth interviews were conducted with 38 Community Midwives (CMWs), 20 policymakers, 45 healthcare providers and 136 community members. A critical policy document review was conducted. National and local level data were brought together. Main outcomes Alignment of programme theory with real-world practice. Results Data revealed gaps between programme theory, assumptions and reality on the ground. The design of the programme failed to take into account: (1) the incongruity between the role of a midwife and dominant class and gendered norms that devalue such a role; (2) market and consumer behaviour that prevented CMWs from establishing private practices; (3) the complexity of public–private sector cooperation. Uniform deployment policies failed to consider existing provider density and geography. Conclusions Greater attention to programme theory and the ‘real-world’ setting during design of maternal health strategies is needed to achieve consistent results in different contexts

Structural basis of PROTAC cooperative recognition for selective protein degradation

Inducing macromolecular interactions with small molecules to activate cellular signaling is a challenging goal. PROTACs (proteolysis-targeting chimeras) are bifunctional molecules that recruit a target protein in proximity to an E3 ubiquitin ligase to trigger protein degradation. Structural elucidation of the key ternary ligase-PROTAC-target species and its impact on target degradation selectivity remain elusive. We solved the crystal structure of Brd4 degrader MZ1 in complex with human VHL and the Brd4 bromodomain (Brd4BD2). The ligand folds into itself to allow formation of specific intermolecular interactions in the ternary complex. Isothermal titration calorimetry studies, supported by surface mutagenesis and proximity assays, are consistent with pronounced cooperative formation of ternary complexes with Brd4BD2. Structure-based-designed compound AT1 exhibits highly selective depletion of Brd4 in cells. Our results elucidate how PROTAC-induced de novo contacts dictate preferential recruitment of a target protein into a stable and cooperative complex with an E3 ligase for selective degradation

PERAN ORANG TUA DALAM MEMBINA NILAI KARAKTER ANAK DI KECAMATAN SIMPANG TIGA ACEH BESAR

ABSTRAKRahmayanti KS, Sri. 2016. Peran Orang Tua Dalam Membina Nilai Karakter Anak di Kecamatan Simpang Tiga Kabupaten Aceh Besar. Skripsi, Jurusan Pendidikan Kesejahteraan Keluarga, Fakultas Keguruan dan Ilmu Pendidikan, Universitas Syiah Kuala. Pembimbing:(1)Dr. Anizar Ahmad, M.Pd., (2). Dra. Fitriana, M.SI.Kata Kunci: Nilai Karakter Anak, Peran Orang TuaPeran orang tua adalah partisipasi atau kesadaran jiwa orang tua untuk memperdulikan anaknya, terutama dalam hal memberikan dan memenuhi kebutuhan hidup anaknya baik dari segi sosial maupun material. Penelitian ini untuk mengetahui usaha orang tua dalam membina nilai karakter anak di Kecamatan Simpang Tiga Kabupaten Aceh Besar. Penelitian ini bertujuan untuk (1) mengetahui usaha yang dilakukan orang tua dalam membina nilai karakter anak dan (2) mengetahui sistem pengawasan yang diterapkan oleh orang tua terhadap anak. Metode yang digunakan dalam penelitian ini adalah metode deskriptif kuantitatif. Data penelitian ini bersumber dari orang tua yang memiliki anak usia 4 sampai 10 tahun berjumlah 28 keluarga, pengumpulan data menggunakan kuisioner. Pengolahan data penelitian ini menggunakan rumus persentase. Simpulan penelitian ini berpengaruh pada usaha orang tua dalam membina nilai karakter anak, walau sebagian kecil yang mengetahui nilai-nilai karakter, tetapi sebagian besar sudah berusaha menanamkan nilai karakter tersebut. Lebih dari setengah responden menanamkan nilai religius kepada anak dengan tujuan agar anak mempunyai akhlak yang mulia kedepannya. Sedangkan yang berperan dalam membina nilai karakter anak adalah suami dan istri. Sistem pengawasan yang diterapkan orang tua berpengaruh terhadap pembentukkan nilai karakter anak. Seluruh anak termasuk kedalam katagori anak yang mudah bersahabat. Penanaman nilai karakter pada anak di mulai pada awal masa kanak-kanak ketika berumur 2-6 tahun. Responden juga menerapkan perilaku disiplin kepada anak karena usia awal kanak-kanak merupakan usia yang masih rentan, dan akan meniru semua yang dikerjakan oleh orang tuanya. Saran untuk orang tua agar dapat mendidik anaknya dengan baik, tidak mengedepankan emosi, dapat meluangkan waktu, adanya komunikasi yang dibina orang tua dengan anak, dan jangan bersikap apatis terhadap apa yang dikerjakan sianak

On the fallibility of human memory for future actions

Human memory is a system that is inherently fallible and prone to distortion, and our memory for future actions is no exception. Prospective memory is defined either as remembering to carry out a task at a particular moment in the future or as the timely execution of a previously formed intention. For a variety of reasons, one may miss this prearranged moment and thus fail to fulfill an intention. This thesis focuses on the factors that may affect the fulfilment of a delayed intention and contribute to prospective memory failures. As the rather scant literature on the effect of stress on prospective memory functioning has produced contradictory findings, Part One of this Thesis investigates the role of stress in prospective memory failures in a strict sense, namely forgetting to carry out intended actions at the appointed time and place. One study involving healthy participants examines the disruptive effect of daily stress on prospective memory functioning and explores the moderating role of individual factors in modulating the harmful consequences associated with stress in everyday life. Another study carried out with healthcare workers investigates how work stress and burnout may contribute to forgetting clinical tasks, which may result in potential adverse events jeopardizing patient safety. Besides stress, misremembering future intentions may also arise from the lingering effect of misinformation on our memory, attitudes, and behaviors. Part Two of this Thesis, encompassing 6 experiments on healthy participants, shows how inaccurate and invalid information survive despite sophisticated correction attempts, influencing memory and reported future intentions. Overall, the results of the studies presented in this Thesis prove the fallibility of our memory for future actions. Various techniques to reduce the risks associated with memory failures are discussed