Enzyme-induced Formation of ß-Lactoglobulin Fibrils by AspN Endoproteinase

This paper describes a low temperature, enzymatic route to induce fibrillar structures in a protein solution. The route comprises two steps. First, ß-lactoglobulin was hydrolyzed into peptides at pH 8 and 37°C with the enzyme AspN endoproteinase, which resulted in the formation of random aggregates. After hydrolysis, the pH was lowered to 2. As a result, long fibrillar aggregates were formed which was observed using transmission electron microscopy and Thioflavin T fluorescence measurements

Onderzoekingen over levenswijze en bestrijding van den witten rijstboorder op Java

For practical control, a study was made of Scirpophaga innotata Wlk., a noxious rice stalk borer in certain regions of Java.Caterpillars normally developed and pupated in stalks that had not reached the stage of ear formation. In paddy from before flowering, however, caterpillars entered into a resting stage (diapause), induced by food rich in carbohydrates and proteins.After harvest diapausing caterpillars remained in the stubble. Further development started after at least 3 months diapause and the stubble and the soil were wetted. This was usually with the first heavy rainfall in the west monsoon. The longer the diapause the sooner moths appeared; with a 3 months' diapause moths emerged 80 days after rainfall began, with a 6 months' diapause after a mean of 33 days.The cultural practice of timed rice sowing was introduced. Sowing started not earlier than one month after the first rains had fallen. Thus infestation of the young fields was prevented for the greater part

Profiling patterns of fecal 20-oxopregnane concentrations during ovarian cycles in free-ranging southern white rhinoceros (Ceratotherium simum simum)

Unlike their wild counterparts, many white rhinoceros females in captivity fail to reproduce successfully such that current captive populations are not self-sustaining. The causes of the problem are poorly understood. Variation in cycle length and long periods of acyclicity are characteristics of the majority of these non-reproducing females in captivity but it is unknown whether these characteristics are a feature of reproductively successful free-ranging females. This study therefore aimed to monitor cyclic activity in a wild population of southern white rhinoceros at Lapalala Wilderness, South Africa, by measuring the concentrations of immunoreactive fecal progestagen metabolites (fPM). Five adult females were tracked twice per week for 20 months and if located a fresh fecal sample was collected. Reproductive events and group structural dynamics were also recorded and subsequently correlated with the fPM data. The baseline concentration of fPM was 0.69 ± 0.20 μg/g DW while concentrations during pregnancy were 30- to 400- fold higher. The females exhibited estrous cycle lengths of 30.6 ± 7.7 days and, based on fPM data, gestation length in one female was 502 ± 3 days. Year-round monitoring showed no clear evidence of seasonality in ovarian activity. During cyclic luteal activity females were often seen in the presence of a dominant bull. One female stopped cycling after removal of the local dominant bull and luteal activity only returned after a new bull was introduced. This suggests that white rhinoceros females in the wild might need external stimuli from a male to ovulate. These findings indicate that the irregular cyclicity reported for white rhinoceros housed in zoos and animal parks may result from conditions in captivity and account for reduced fertility.The University of Western Australia, the Mammal Research Institute and Endocrine Research Laboratory, University of Pretoria, the Institute for Breeding Rare and Endangered African Mammals, the AAZK‘s Bowling For Rhinos, and the SAVE African Rhino Foundation.http://www.elsevier.com/locate/anireprosci2016-10-31hb201

CMG2/ANTXR2 regulates extracellular collagen VI which accumulates in hyaline fibromatosis syndrome.

Loss-of-function mutations in capillary morphogenesis gene 2 (CMG2/ANTXR2), a transmembrane surface protein, cause hyaline fibromatosis syndrome (HFS), a severe genetic disorder that is characterized by large subcutaneous nodules, gingival hypertrophy and severe painful joint contracture. Here we show that CMG2 is an important regulator of collagen VI homoeostasis. CMG2 loss of function promotes accumulation of collagen VI in patients, leading in particular to nodule formation. Similarly, collagen VI accumulates massively in uteri of Antxr2 <sup>-/-</sup> mice, which do not display changes in collagen gene expression, and leads to progressive fibrosis and sterility. Crossing Antxr2 <sup>-/-</sup> with Col6a1 <sup>-/-</sup> mice leads to restoration of uterine structure and reversion of female infertility. We also demonstrate that CMG2 may act as a signalling receptor for collagen VI and mediates its intracellular degradation

Evaluating Surveillance Strategies for the Early Detection of Low Pathogenicity Avian Influenza Infections

In recent years, the early detection of low pathogenicity avian influenza (LPAI) viruses in poultry has become increasingly important, given their potential to mutate into highly pathogenic viruses. However, evaluations of LPAI surveillance have mainly focused on prevalence and not on the ability to act as an early warning system. We used a simulation model based on data from Italian LPAI epidemics in turkeys to evaluate different surveillance strategies in terms of their performance as early warning systems. The strategies differed in terms of sample size, sampling frequency, diagnostic tests, and whether or not active surveillance (i.e., routine laboratory testing of farms) was performed, and were also tested under different epidemiological scenarios. We compared surveillance strategies by simulating within-farm outbreaks. The output measures were the proportion of infected farms that are detected and the farm reproduction number (Rh). The first one provides an indication of the sensitivity of the surveillance system to detect within-farm infections, whereas Rh reflects the effectiveness of outbreak detection (i.e., if detection occurs soon enough to bring an epidemic under control). Increasing the sampling frequency was the most effective means of improving the timeliness of detection (i.e., it occurs earlier), whereas increasing the sample size increased the likelihood of detection. Surveillance was only effective in preventing an epidemic if actions were taken within two days of sampling. The strategies were not affected by the quality of the diagnostic test, although performing both serological and virological assays increased the sensitivity of active surveillance. Early detection of LPAI outbreaks in turkeys can be achieved by increasing the sampling frequency for active surveillance, though very frequent sampling may not be sustainable in the long term. We suggest that, when no LPAI virus is circulating yet and there is a low risk of virus introduction, a less frequent sampling approach might be admitted, provided that the surveillance is intensified as soon as the first outbreak is detected

Faecal progestagen profiles in wild southern white rhinoceros (Ceratotherium simum simum)

Knowledge of the reproductive biology of wild animals can provide valuable information for the development of appropriate in situ and ex situ management plans. The present study aimed to establish a non-invasive protocol for monitoring faecal progestagen metabolite (FPM) patterns in wild female southern white rhinoceroses (Ceratotherium simum simum). Six adult females at Lapalala Wilderness, South Africa, were tracked and accurately identified at least once every week. Three animals gave birth during the study period. Fresh faecal samples were collected for 12 months and stored frozen at –20°C until analysis with an enzyme immunoassay utilizing an antiserum raised against 5 -pregnan-3 -ol-20-one which cross-reacts with a number of progestagens. Mean FPM concentrations were 35 to 64-fold higher during pregnancy (55–145 days before parturition) compared to postpartum (120–140 days after parturition) (P < 0.001). Also, the non-pregnant animals had mean FPM concentrations significantly higher than postpartum values (P = 0.006). Our results show that non-invasive FPM measurements provide information on the pregnancy status of wild female white rhinoceroses, and may be used for the detection of pregnancy in free-living individuals, without the necessity of immobilization and/or relocation of the animal. This information has potential value for optimizing breeding management of wild and captive populations.We wish to thank the Institute for Breeding Rare and Endangered African Mammals (IBREAM) for making this research possible.We are indebted to the following students from the faculty of Veterinary Medicine at Utrecht University for their contribution in collecting faecal samples: Yvette Charbon, Belinda Bitter, Meike Meijers and Julie Swinkels. We are especially thankful to Thomas Litshani, whose exceptional rhinoceros knowledge and tracking skills greatly contributed to the number of samples collected.We are most grateful to the management of the Lapalala Wilderness reserve: Duncan Parker, Mike Gregor, Anton Walker, Roger Collinson and Anthony and Erin Roberts for permission to conduct research in the reserve as well as for their help and support. We also thank John Hanks and Henk Bertschinger for their encouragement and helpful advice, and Stefanie Ganswindt for expert help in laboratory techniques.http://africanzoology.journals.ac.za/am2013ab201

Relative cerebral flow from dynamic PIB scans as an alternative for FDG scans in Alzheimer's disease PET studies

In Alzheimer's Disease (AD) dual-tracer positron emission tomography (PET) studies with 2-[F-18]-fluoro-2-deoxy-D-glucose (FDG) and C-11-labelled Pittsburgh Compound B (PIB) are used to assess metabolism and cerebral amyloid-beta deposition, respectively. Regional cerebral metabolism and blood flow (rCBF) are closely coupled, both providing an index for neuronal function. The present study compared PIB-derived rCBF, estimated by the ratio of tracer influx in target regions relative to reference region (R-1) and early-stage PIB uptake (ePIB), to FDG scans. Fifteen PIB positive (+) patients and fifteen PIB negative (-) subjects underwent both FDG and PIB PET scans to assess the use of R-1 and ePIB as a surrogate for FDG. First, subjects were classified based on visual inspection of the PIB PET images. Then, discriminative performance (PIB+ versus PIB-) of rCBF methods were compared to normalized regional FDG uptake. Strong positive correlations were found between analyses, suggesting that PIB-derived rCBF provides information that is closely related to what can be seen on FDG scans. Yet group related differences between method's distributions were seen as well. Also, a better correlation with FDG was found for R-1 than for ePIB. Further studies are needed to validate the use of R-1 as an alternative for FDG studies in clinical applications

Protection and Virus Shedding of Falcons Vaccinated against Highly Pathogenic Avian Influenza A Virus (H5N1)

Virus shedding by vaccinated birds was markedly reduced

Functional Diversity and Structural Disorder in the Human Ubiquitination Pathway

The ubiquitin-proteasome system plays a central role in cellular regulation and protein quality control (PQC). The system is built as a pyramid of increasing complexity, with two E1 (ubiquitin activating), few dozen E2 (ubiquitin conjugating) and several hundred E3 (ubiquitin ligase) enzymes. By collecting and analyzing E3 sequences from the KEGG BRITE database and literature, we assembled a coherent dataset of 563 human E3s and analyzed their various physical features. We found an increase in structural disorder of the system with multiple disorder predictors (IUPred - E1: 5.97%, E2: 17.74%, E3: 20.03%). E3s that can bind E2 and substrate simultaneously (single subunit E3, ssE3) have significantly higher disorder (22.98%) than E3s in which E2 binding (multi RING-finger, mRF, 0.62%), scaffolding (6.01%) and substrate binding (adaptor/substrate recognition subunits, 17.33%) functions are separated. In ssE3s, the disorder was localized in the substrate/adaptor binding domains, whereas the E2-binding RING/HECT-domains were structured. To demonstrate the involvement of disorder in E3 function, we applied normal modes and molecular dynamics analyses to show how a disordered and highly flexible linker in human CBL (an E3 that acts as a regulator of several tyrosine kinase-mediated signalling pathways) facilitates long-range conformational changes bringing substrate and E2-binding domains towards each other and thus assisting in ubiquitin transfer. E3s with multiple interaction partners (as evidenced by data in STRING) also possess elevated levels of disorder (hubs, 22.90% vs. non-hubs, 18.36%). Furthermore, a search in PDB uncovered 21 distinct human E3 interactions, in 7 of which the disordered region of E3s undergoes induced folding (or mutual induced folding) in the presence of the partner. In conclusion, our data highlights the primary role of structural disorder in the functions of E3 ligases that manifests itself in the substrate/adaptor binding functions as well as the mechanism of ubiquitin transfer by long-range conformational transitions. © 2013 Bhowmick et al

Late Endosomal Cholesterol Accumulation Leads to Impaired Intra-Endosomal Trafficking

Background Pathological accumulation of cholesterol in late endosomes is observed in lysosomal storage diseases such as Niemann-Pick type C. We here analyzed the effects of cholesterol accumulation in NPC cells, or as phenocopied by the drug U18666A, on late endosomes membrane organization and dynamics. Methodology/Principal Findings Cholesterol accumulation did not lead to an increase in the raft to non-raft membrane ratio as anticipated. Strikingly, we observed a 2–3 fold increase in the size of the compartment. Most importantly, properties and dynamics of late endosomal intralumenal vesicles were altered as revealed by reduced late endosomal vacuolation induced by the mutant pore-forming toxin ASSP, reduced intoxication by the anthrax lethal toxin and inhibition of infection by the Vesicular Stomatitis Virus. Conclusions/Significance These results suggest that back fusion of intralumenal vesicles with the limiting membrane of late endosomes is dramatically perturbed upon cholesterol accumulation