Sex ratio estimations of Chelonia mydas hatchlings at Samandağ Beach, Turkey

Samandağ Beach, an important nesting location for endangered green turtles (Chelonia mydas), is located at the most eastern part of the Mediterranean in Turkey. Hatchling sex ratio can be one of the key factors for green turtle protection, which is under the threat of global climate change. In this study, sex ratios of hatchlings were estimated by histological examination of dead hatchlings (n = 190) and by nest temperatures (n = 14) at Samandağ Beach between the 2003 and 2007 nesting seasons. The histological examination of dead C. mydas hatchlings and temperature-recorded nests indicated a female-biased sex ratio on Samandağ Beach. By histological examination, top and bottom levels of nests were recorded as 94% and 64% of females. Sex ratios during the middle third of the incubation period ranged from 39% to 97% for females with a mean of 74%. Distance from the sea was correlated with the middle third of incubation temperature (r = –0.604, P = 0.022) and female ratio (r = –0.573, P = 0.032). While nest temperatures and nest parameters such as hatching success, clutch size, nest distance from the sea and from vegetation, nest depth, and the day of emergence were not found significantly different among years (P > 0.05), incubation duration was found significantly different among years (P < 0.05). The results showed a female-dominated sex ratio by both histology and nest temperature. The knowledge of hatchling sex ratios provides information on demographic parameters, and coupled with appropriate conservation measures this can make important contributions to studies of climate change effects on green sea turtle populations. © TÜBİTAK

Ubiquitin-based probes prepared by total synthesis to profile the activity of deubiquitinating enzymes

Epitope-tagged active-site-directed probes are widely used to visualize the activity of deubiquitinases (DUBs) in cell extracts, to investigate the specificity and potency of small-molecule DUB inhibitors, and to isolate and identify DUBs by mass spectrometry. With DUBs arising as novel potential drug targets, probes are required that can be produced in sufficient amounts and to meet the specific needs of a given experiment. The established method for the generation of DUB probes makes use of labor-intensive intein-based methods that have inherent limitations concerning the incorporation of unnatural amino acids and the amount of material that can be obtained. Here, we describe the total chemical synthesis of active-site-directed probes and their application to activity-based profiling and identification of functional DUBs. This synthetic methodology allowed the easy incorporation of desired tags for specific applications, for example, fluorescent reporters, handles for immunoprecipitation or affinity pull-down, and cleavable linkers. Additionally, the synthetic method can be scaled up to provide significant amounts of probe. Fluorescent ubiquitin probes allowed faster, in-gel detection of active DUBs, as compared to (immuno)blotting procedures. A biotinylated probe holding a photocleavable linker enabled the affinity pull-down and subsequent mild, photorelease of DUBs. Also, DUB activity levels were monitored in response to overexpression or knockdown, and to inhibition by small molecules. Furthermore, fluorescent probes revealed differential DUB activity profiles in a panel of lung and prostate cancer cells

Variation of mohair staple length across Angora goat fleeces : implications for animal selection and fleece evaluation

The present study aimed to determine how the average mohair staple length (SL) differences between nine sampling sites vary between sex and flock, to identify differences in SL variability between sampling sites as a result of between-animal and between-sire variability and to determine SL correlations between sampling sites in between-animal and between-sire variability. Australian Angora goats (n=301) from two farms in southern Australia were sampled at 12 and 18 months of age at nine sites (mid side, belly, brisket, hind flank, hip, hock, mid back, neck and shoulder). Staples were taken prior to shearing at skin level and stretched SL determined. For each shearing, differences in SL between sampling sites, how these differences were affected by farm, sex and sire, and the covariance between sites for sire and individual animal effects were investigated by restricted maximum likelihood (REML) analyses. The median mid-side SL at 12 and 18 months of age was 110 and 130 mm, respectively, but the actual range in mid-side SL was 65&ndash;165 mm. There was an anterior&ndash;posterior decline in SL with the hock being particularly short. There was no evidence that the between-site correlation of the sire effects differed from 1, indicating that genetic selection for SL at one site will be reflected in SL over the whole fleece. However, low heritabilities of SL at the hock, belly and brisket or at any site at 12 months of age were obtained. There was more variability between sites than between sires, but the between-animal variation was greater. The hip and mid-back sites can be recommended for within-flock (culling) and genetic selection for SL due to their low sampling variability, moderate heritability and ease of location. <br /

Ubiquitinome Profiling Reveals <i>in Vivo</i> UBE2D3 Targets and Implicates UBE2D3 in Protein Quality Control

Ubiquitination has crucial roles in many cellular processes, and dysregulation of ubiquitin machinery enzymes can result in various forms of pathogenesis. Cells only have a limited set of ubiquitin-conjugating (E2) enzymes to support the ubiquitination of many cellular targets. As individual E2 enzymes have many different substrates and interactions between E2 enzymes and their substrates can be transient, it is challenging to define all in vivo substrates of an individual E2 and the cellular processes it affects. Particularly challenging in this respect is UBE2D3, an E2 enzyme with promiscuous activity in vitro but less defined roles in vivo. Here, we set out to identify in vivo targets of UBE2D3 by using stable isotope labeling by amino acids in cell culture–based and label-free quantitative ubiquitin diGly proteomics to study global proteome and ubiquitinome changes associated with UBE2D3 depletion. UBE2D3 depletion changed the global proteome, with the levels of proteins from metabolic pathways, in particular retinol metabolism, being the most affected. However, the impact of UBE2D3 depletion on the ubiquitinome was much more prominent. Interestingly, molecular pathways related to mRNA translation were the most affected. Indeed, we find that ubiquitination of the ribosomal proteins RPS10 and RPS20, critical for ribosome-associated protein quality control, is dependent on UBE2D3. We show by Targets of Ubiquitin Ligases Identified by Proteomics 2 methodology that RPS10 and RPS20 are direct targets of UBE2D3 and demonstrate that the catalytic activity of UBE2D3 is required to ubiquitinate RPS10 in vivo. In addition, our data suggest that UBE2D3 acts at multiple levels in autophagic protein quality control. Collectively, our findings show that depletion of an E2 enzyme in combination with quantitative diGly-based ubiquitinome profiling is a powerful tool to identify new in vivo E2 substrates, as we have done here for UBE2D3. Our work provides an important resource for further studies on the in vivo functions of UBE2D3.Dutch Ministry of Health KWF-NKI2012- 5305Dutch Cancer Society 11369/2017-

Valorisation of rice husks using a TORBED® combustion process

World production of rice exceeds 750 million tonnes per year of which a fifth is removed in the form of rice husk during the milling process. The use of rice husks as a source of sustainable and renewable energy is often hindered by lack of capital and a poor understanding of rice husk combustion characteristics. This results in the selection of poor quality technology which generates significant quantities of harmful crystalline silica waste. Despite previous work in the area, detailed characterisation of the combustion of rice husk ash in a TORBED reactor across a wide temperature range has not yet been attempted and little effort has been directed towards assessing the economic viability of generating quality rice husk ashes. The use of a TORBED reactor enables low residual carbon after combustion without the generation of harmful crystalline material. Rice husk was combusted in a 400 mm reactor at temperatures between 700 and 950 °C. In the subsequent characterisation studies the resulting materials were shown to be fully amorphous high purity silica (&gt; 95%) and were readily digested in a series of alkaline digestion experiments. Complete silica conversion was only possible using uneconomic Na₂O/SiO₂ ratios and further optimisation of the combustion process to generate higher surface area material is necessary to increase the digestion rates further. Provisional economic analysis suggests that sales of the by-product enhance the returns from rice husk based power generation. TORBED reactors enable the combustion of rice husk with considerable operating flexibility and they generate products that could be used to displace resource intensive products and processes thus, added value from the by-products can be obtained by using TORBED reactor technology

Self-determined citizens? New forms of civic activism and citizenship in Armenia

This article examines the recent emergence and growth of grassroots social movements in Armenia which are locally known as ‘civic initiatives’. It considers what their emergence tells us about the development of civil society and the changing understandings and practices of citizenship in Armenia in the post-Soviet period. It analyses why civic initiatives explicitly reject and distance themselves from formal, professionalised NGOs and what new models of civic activism and citizenship they have introduced. It argues that civic initiatives embrace a more political understanding of civil society than that which was introduced by Western donors in the 1990s