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In the early secretory pathway, endoplasmic reticulum (ER) and Golgi membranes form a nearly spherical interface. In this ribosome-excluding zone, bidirectional transport of cargo coincides with a spatial segregation of anterograde and retrograde carriers by an unknown mechanism. We show that at physiological conditions, the Trk-fused gene (TFG) self-organizes to form a hollow, anisotropic condensate that matches the dimensions of the ER–Golgi interface and is dynamically regulated across the cell cycle. Regularly spaced hydrophobic residues in TFG control the condensation mechanism and result in a porous condensate surface. We find that TFG condensates act as a molecular sieve capable of allowing access of anterograde coats (COPII) to the condensate interior while restricting retrograde coats (COPI). We propose that a hollow TFG condensate structures the ER–Golgi interface to create a diffusion-limited space for anterograde transport. We further propose that TFG condensates optimize membrane flux by insulating secretory carriers in their lumen from retrograde carriers outside TFG cages. The authors show that protein TFG self-organizes to form a hollow sphere that helps to guide and spatially organize bidirectional transport at a critical junction between the ER and Golgi in the early secretory pathway.

Coronatin-2, a 14.5 kDa protein, was isolated from culture filtrates of the entomopathogenic fungus Conidiobolus coronatus (Costantin) Batko (Entomophthoramycota: Entomophthorales). After LC–MS/MS (liquid chromatography tandem mass spectrometry) analysis of the tryptic peptide digest of coronatin-2 and a mass spectra database search no orthologs of this protein could be found in fungi. The highest homology was observed to the partial translation elongation factor 1a from Sphaerosporium equinum (protein sequence coverage, 21%), with only one peptide sequence, suggesting that coronatin-2 is a novel fungal protein that has not yet been described. In contrast to coronatin-1, an insecticidal 36 kDa protein, which shows both elastolytic and chitinolytic activity, coronatin-2 showed no enzymatic activity. Addition of coronatin-2 into cultures of hemocytes taken from larvae of Galleria mellonella Linnaeus (Lepidoptera: Pyralidae), resulted in progressive disintegration of nets formed by granulocytes and plasmatocytes due to rapid degranulation of granulocytes, extensive vacuolization of plasmatocytes accompanied by cytoplasm expulsion, and cell disintegration. Spherulocytes remained intact, while oenocytes rapidly disintegrated. Coronatin-2 produced 80% mortality when injected into G. mellonella at 5 µg larva−1. Further study is warranted to determine the relevance of the acute toxicity of coronatin-2 and its effects on hemocytes in vitro to virulence of C. coronatus against its hosts.

Concurrent engineering through overlapping of activities (i.e., fast-tracking) has been used as a schedule acceleration technique. Fast-track construction projects are generally recognized as riskier and subject to risks arising due to the concurrency of work. This work reports the risk perception of construction professionals to three different degrees of overlapping. Semi-structured interviews were used to collect the data, and the analysis applied data transformation and descriptive statistics. The risks were mainly perceived in the middle degree of overlapping and in activities occurring earlier in the schedule. The low and high degrees of overlapping were mainly perceived as having no risk or not being feasible, respectively. The four risk types accounted for most of the perceived threats: construction error, design change, crew interference, and poor construction productivity. The findings of this study suggest that construction professionals perceive risks differently based on the amount of activity overlapping. It is consistent with previous studies that found that risks decrease as the project advances and that fast-track projects face additional risks.

•A novel dual route vaccination using the MERS-CoV RBD sub-unit has been developed.•Murine antisera induced to the RBD protein , were neutralising in vitro.•MERS-CoV susceptibility was induced in naïve mice with Ad5hDPP4.•Passive transfer of anti-RBD sera protected susceptible mice.•Protected mice had a significantly reduced viral titre (P = 0.02) in their lungs. Murine antisera with neutralising activity for the coronavirus causative of Middle East respiratory syndrome (MERS) were induced by immunisation of Balb/c mice with the receptor binding domain (RBD) of the viral Spike protein. The murine antisera induced were fully-neutralising in vitro for two separate clinical strains of the MERS coronavirus (MERS-CoV). To test the neutralising capacity of these antisera in vivo, susceptibility to MERS-CoV was induced in naive recipient Balb/c mice by the administration of an adenovirus vector expressing the human DPP4 receptor (Ad5-hDPP4) for MERS-CoV, prior to the passive transfer of the RBD-specific murine antisera to the transduced mice. Subsequent challenge of the recipient transduced mice by the intra-nasal route with a clinical isolate of the MERS-CoV resulted in a significantly reduced viral load in their lungs, compared with transduced mice receiving a negative control antibody. The murine antisera used were derived from mice which had been primed sub-cutaneously with a recombinant fusion of RBD with a human IgG Fc tag (RBD-Fc), adsorbed to calcium phosphate microcrystals and then boosted by the oral route with the same fusion protein in reverse micelles. The data gained indicate that this dual-route vaccination with novel formulations of the RBD-Fc, induced systemic and mucosal anti-viral immunity with demonstrated in vitro and in vivo neutralisation capacity for clinical strains of MERS-CoV.

The construction industry has extensively applied the fast-tracking approach to the demanding need for the fast delivery of infrastructure projects. However, the fast-track strategy might be threatened by distinctive risks or changes in risk characteristics that emerge when activities are overlapped (overlapping risks). This article proposes a risk assessment simulation model to quantify the economic impact of overlapping risks on fast-track infrastructure projects. The model uses Monte Carlo simulation and a proprietary engine solution for the optimization procedure. It quantifies the overlapping risk impacts in the project duration and cost that could originate in three different overlapping degrees and evaluates the optimal overlapping degree to reduce the impact of the overlapping risks. The model demonstration used a commercial renovation project. The results suggest that overlapping risks have a high potential impact on the total cost, although with a high probability of attaining the target duration. Eight top risks affected the total duration, cost, or both. The optimum overlapping to reduce the economic impact and achieve the target project duration combines different overlapping degrees. This study contribution is a model for fast-track projects considering overlapping risks, their impact characteristic as a distribution, and the potential relationship between these risks.

•Induction of rapid immunity (d35); median serum IgG titres to F1 and V of >1800 and >2200 µg/ml respectively.•Induction of specific IgA in serum and faecal extracts.•Full protective efficacy against 2 × 104 median lethal doses of Y. pestis (s.c.) at day 46, 14 days after single boost.•Oral dose effectively boosts systemic immunity and induced mucosal immunity.•VypVaxDuo fulfils rapid response criteria for a vaccine designed for emergency use authorisation. Here, we report a dual-route vaccination approach for plague, able to induce a rapid response involving systemic and mucosal immunity, whilst also providing ease of use in those resource-poor settings most vulnerable to disease outbreaks. This novel vaccine (VypVaxDuo) comprises the recombinant F1 and V proteins in free association. VypVaxDuo has been designed for administration via a sub-cutaneous priming dose followed by a single oral booster dose and has been demonstrated to induce early onset immunity 14 days after the primary immunisation; full protective efficacy against live organism challenge was achieved in Balb/c mice exposed to 2 × 104 median lethal doses of Yersinia pestis Co92, by the sub-cutaneous route at 25 days after the oral booster immunisation. This dual-route vaccination effectively induced serum IgG and serum and faecal IgA, specific for F1 and V, which constitute two key virulence factors in Y. pestis, and is therefore suitable for further development to prevent bubonic plague and for evaluation in models of pneumonic plague. This is an essential requirement for control of disease outbreaks in areas of the world endemic for plague and is supported further by the observed exceptional stability of the primary vaccine formulation in vialled form under thermostressed conditions (40 °C for 29 weeks, and 40 °C with 75% relative humidity for 6 weeks), meaning no cold chain for storage or distribution is needed. In clinical use, the injected priming dose would be administered on simple rehydration of the dry powder by means of a dual barrel syringe, with the subsequent single booster dose being provided in an enteric-coated capsule suitable for oral self-administration.

Non-mammal models are gaining popularity due to their ease of maintenance, low cost and lack of ethical restrictions. As some processes taking place in insect hemocytes are mirrored in human immunocytes, many immunological studies are now turning to the use of insect models like Galleria mellonella. G. mellonella larvae are susceptible to infection by Conidiobolus coronatus: an entomopathogenic fungus causing rhinofacial mycosis in humans. The study examines G. mellonella larvae during C. coronatus infection for the presence of certain compounds known to be involved in the human immunological response (histamine, HSF1, Cysteinyl leukotriene TLR1 and TLR2). G. mellonella larvae were exposed to fully grown fungus, and the results were determined by ELISA tests, fluorescence microscopy and flow cytometry. CysLT showed no significant changes between the control group and the infected larvae. Both histamine and HSF1 levels increased significantly after exposure to C. coronatus. Immunolocalization and flow cytometry of TLR1 and TLR2 proves that both receptors increase their levels due to the fungus infection. Our findings show that the cellular response of the G. mellonella is similar to the human response, indicating that G. mellonella larvae may be considered as a human-like immunological model in further studies.

Roadway pavement surface distress information is critical for effective pavement asset management, and subsequently, transportation management agencies at all levels (i.e., federal, state, and local) dedicate a large amount of time and money to routinely evaluate pavement surface distress conditions as the core of their asset management programs. However, currently adopted ground-based evaluation methods for pavement surface conditions have many disadvantages, like being time-consuming and expensive. Aircraft-based evaluation methods, although getting more attention, have not been used for any operational evaluation programs yet because the acquired images lack the spatial resolution to resolve finer scale pavement surface distresses. Hyper-spatial resolution natural color aerial photography (HSR-AP) provides a potential method for collecting pavement surface distress information that can supplement or substitute for currently adopted evaluation methods. Using roadway pavement sections located in the State of New Mexico as an example, this research explored the utility of aerial triangulation (AT) technique and HSR-AP acquired from a low-altitude and low-cost small-unmanned aircraft system (S-UAS), in this case a tethered helium weather balloon, to permit characterization of detailed pavement surface distress conditions. The Wilcoxon Signed Rank test, Mann-Whitney U test, and visual comparison were used to compare detailed pavement surface distress rates measured from HSR-AP derived products (orthophotos and digital surface models generated from AT) with reference distress rates manually collected on the ground using standard protocols. The results reveal that S-UAS based hyper-spatial resolution imaging and AT techniques can provide detailed and reliable primary observations suitable for characterizing detailed pavement surface distress conditions comparable to the ground-based manual measurement, which lays the foundation for the future application of HSR-AP for automated detection and assessment of detailed pavement surface distress conditions.

The composition of cuticular and internal n-alkanes in Lucilia sericata larvae, pupae, and male and female imagines were studied. The cuticular and internal lipid extracts were separated by HPLC-LLSD, after which the hydrocarbon fraction was identified by GC/MS in selected ion monitoring (SIM) and total ion current (TIC) modes. The cuticular lipids of the larvae contained seven n-alkanes from C23 to C31. The major n-alkane in L. sericata larvae was C29 (42.1%). The total cuticular n-alkane content in the cuticular lipids was 31.46 μg g−1 of the insect body. The internal lipids of L. sericata larvae contained five n-alkanes ranged from C25 to C31. The most abundant compound was C27 (61.71 μg g−1 of the insect body). Eighteen n-alkanes from C14 to C31 were identified in the cuticular lipids of the pupae. The most abundant n-alkanes ranged from C25 to C31; those with odd-numbered carbon chains were particularly abundant, the major one being C29:0 (59.5%). Traces of eight cuticular n-alkanes were present. The internal lipids of L. sericata pupae contained five n-alkanes, ranging from C25 to C31. The cuticular lipids of female imagines contained 17 n-alkanes from C12 to C30. Among the cuticular n-alkanes of females, C27 (47.5%) was the most abundant compound. Four n-alkanes, with only odd-numbered carbon chains, were identified in the internal lipids of females. The lipids from both sexes of L. sericata had similar n-alkane profiles. The cuticular lipids of adult males contained 16 n-alkanes ranging from C13 to C31. C27 (47.9%) was the most abundant cuticular n-alkanes in males. The same n-alkanes only with odd-numbered carbon chains and in smaller quantities of C27 (0.1%) were also identified in the internal lipids of males. The highest amounts of total cuticular n-alkanes were detected in males and females of L. sericata (330.4 and 158.93 μg g−1 of the insect body, respectively). The quantities of total cuticular alcohols in larvae and pupae were smaller (31.46 μg g−1 and 42.08 μg g−1, respectively). The internal n-alkane contents of larvae, pupae, and male and female imagines were significantly higher than the cuticular n-alkane contents (153.53, 99.60, 360.06 and 838.76 μg g−1 of the insect body, respectively).

To examine the impacts on food and nutrition-related outcomes resulting from participation in urban gardens, especially on healthy food practices, healthy food access, and healthy food beliefs, knowledge and attitudes. The systematic review identified studies by searching the PubMed, ERIC, LILACS, Web of Science and Embase databases. An assessment of quality and bias risk of the studies was carried out and a narrative summary was produced. Studies published as original articles in peer-reviewed scientific journals in English, Spanish or Portuguese between 2005 and 2015 were included. The studies included were based on data from adult participants in urban gardens. Twenty-four studies were initially selected based on the eligibility criteria, twelve of which were included. There was important heterogeneity of settings, population and assessment methods. Assessment of quality and bias risk of the studies revealed the need for greater methodological rigour. Most studies investigated community gardens and employed a qualitative approach. The following were reported: greater fruit and vegetable consumption, better access to healthy foods, greater valuing of cooking, harvest sharing with family and friends, enhanced importance of organic production, and valuing of adequate and healthy food. Thematic patterns related to adequate and healthy food associated with participation in urban gardens were identified, revealing a positive impact on practices of adequate and healthy food and mainly on food perceptions.