COPII collar defines the boundary between ER and ER exit site and does not coat cargo containers

COPII and COPI mediate the formation of membrane vesicles translocating in opposite directions within the secretory pathway. Live-cell and electron microscopy revealed a novel mode of function for COPII during cargo export from the ER. COPII is recruited to membranes defining the boundary between the ER and ER exit sites, facilitating selective cargo concentration. Using direct observation of living cells, we monitored cargo selection processes, accumulation, and fission of COPII-free ERES membranes. CRISPR/Cas12a tagging, the RUSH system, and pharmaceutical and genetic perturbations of ER-Golgi transport demonstrated that the COPII coat remains bound to the ER–ERES boundary during protein export. Manipulation of the cargo-binding domain in COPII Sec24B prohibits cargo accumulation in ERES. These findings suggest a role for COPII in selecting and concentrating exported cargo rather than coating Golgi-bound carriers. These findings transform our understanding of coat proteins’ role in ER-to-Golgi transport.     

Comparative life cycle sustainability assessment of urban water reuse at various centralization scales

Purpose

Population growth and urbanization lead to increasing water demand, putting significant pressure on natural water sources. The rising amounts of domestic wastewater (WW) in urban areas may be treated to serve as an alternative water source that may alleviate this pressure. This study examines sustainability of utilizing reclaimed domestic wastewater in urban households for toilet flushing and garden irrigation. It models a city characterized by water scarcity, using a coal-based electricity mix.

Methods

Four approaches were compared: (0) Business-as-usual (BAU) alternative, where the central WW treatment plant effluent is discharged to nature; (1) central WW treatment and urban reuse of the effluent produced; (2) semi-distributed greywater treatment and reuse, at cluster scale; (3) Distributed greywater treatment and reuse, at building scale. Environmental life cycle assessment (LCA), social LCA (S-LCA), and life cycle costing (LCC) were applied to the system model of the above scenarios, with seawater desalination as the source for potable water. System boundaries include water supply, WW collection, and treatment facilities. Analytical hierarchy process (AHP), a multi-criteria decision analysis (MCDA) methodology, was integrated into the life cycle sustainability assessment (LCSA) framework as a means for weighting sustainability criteria through judgment elicitation from a panel of 20 experts.

Results and discussion

Environmentally and socially, the two distributed alternatives perform better in most impact categories. Socially, semi-distributed (cluster scale) reuse is somewhat advantageous over the fully distributed alternative (building scale), due to the benefits of community engagement. Economically, the cluster-level scenario is the most preferable, while the building-scale scenario is the least preferable. A hierarchical representation of the problem’s criteria was constructed, according to the principals of AHP. Each criterion was weighted and those of extreme low importance were eliminated, while maintaining the integrity of the experts’ judgments. Weighted and aggregated sustainability scores revealed that cluster level reclamation, under modeled conditions, is the most sustainable option and the BAU scenario is the least sustainable. The other two alternatives, centralized and fully distributed reclamation, obtained similar intermediate scores.

Conclusions

Distributed urban water reuse was found to be more sustainable than current practice. Different alternative solutions are advantageous in different ways, but overall, the reclamation and reuse of greywater at the cluster level seems to be the best option among the three reuse options examined in this assessment. AHP proved an effective method for aggregating the multiple sustainability criteria. The hierarchical view maintains transparency of all local weights while leading to the final weight vector.

     

Epigenetic metaphors: an interdisciplinary translation of encoding and decoding

Looking at the new and often disputed science of epigenetics, we examined the challenges faced by scientists when they communicate scientific research to the public. We focused on the use of metaphors to illustrate notions of epigenetics and genetics. We studied the “encoding” by epigeneticists and “decoding” in focus groups with diverse backgrounds. We observed considerable overlap in the dominant metaphors favored by both researchers and the lay public. However, the groups differed markedly in their interpretations of which metaphors aided understanding or not. We conclude by discussing the role of metaphors and their interpretations in the context of a shift from pre-deterministic genomic metaphors to more active, dynamic and nuanced epigenetic metaphors. These reflections on the choice of metaphors and differences in encoding/decoding are important for science communication and scientific boundary-maintenance.     

The sentineling—Foraging trade‐off in dominant and subordinate Arabian babblers

Many cooperative breeders forage under predation risks, sentineling is a central activity, and groupmates have to balance between sentineling and foraging. The optimal balance between sentinel activity and foraging may differ among dominant and subordinate individuals, as dominants are more efficient foragers. Two theoretical models pertain to this balance and predict when individuals with different foraging abilities should switch between the two activities on the basis of their energetic state. In one of these models, individuals must attain a critical energetic level by dusk to pass the night, and in the second model fitness is monotonically increasing with the energetic state. We tested these models in the cooperatively breeding Arabian babbler, Turdoides squamiceps. We measured the length of sentinel bouts and the gaps between them both in natural conditions and following experimental feeding. Following feeding ad libitum, subordinates expanded their sentinel bouts significantly more than dominants in comparison with natural conditions. These findings are consistent with the first model, but not with the second. In the experiment, we measured the mass of mealworms consumed by each individual following a sentinel bout relative to its body mass. This ratio was larger for subordinates, indicating that they ended their sentinel bouts at a lower energetic state than dominants. This finding is consistent with the second model, but not with the first. Immediately after eating ad libitum, in 62% of the cases the first behavior performed by the babblers was a new sentinel bout, but in 17% it was a mutual interaction with a groupmate, indicating that social interactions also play a role in the trade‐off vis‐à‐vis sentinel activity.     

Glycerol triacetate as solvent and acyl donor in the production of isoamyl acetate with Candida antarctica lipase B

Glycerol triacetate was successfully used as a green solvent and as the acyl donor in the transesterification of isoamyl alcohol to produce isoamyl acetate using free and immobilized Candida antarctica lipase B. Immobilized lipase was more catalytically active than free lipase and could be easily separated from the reaction mixture by filtration. In addition, it was found that increasing either the reaction temperature or the enzyme to substrate ratio increased the conversion of isoamyl alcohol. Using triacetin as the solvent also enabled the separation of product by simple extraction with petroleum ether and catalyst recycling.     

Fracture mechanics analysis of vertical root fracture from condensation of gutta-percha

A two-dimensional fracture mechanics analysis of vertical root fracture (VRF) in single-canal roots from apical condensation of gutta-percha (gp) is developed. The resulting analytic relation for apical load causing VRF agrees with major trends reported in in-vitro tests on roots subjected to either continuous or, the more clinically relevant, repeating vertical condensation of gp. The model explicitly exposes the role of root canal morphology and dentin fracture toughness on VRF. Ovoid and irregular canals are prone to fracture while the effect of mean root canal radius is modest. Canal taper and instrumentation details may affect VRF only marginally and indirectly. The model predicts dentinal cracks to occur following root canal instrumentation and obturation, which may pose long-term threats to tooth integrity.     

A p38 MAPK-CREB pathway functions to pattern mesoderm in Xenopus

Dorsal-ventral patterning is specified by signaling centers secreting antagonizing morphogens that form a signaling gradient. Yet, how morphogen gradient is translated intracellularly into fate decisions remains largely unknown. Here, we report that p38 MAPK and CREB function along the dorsal-ventral axis in mesoderm patterning. We find that the phosphorylated form of CREB (S133) is distributed in a gradient along the dorsal-ventral mesoderm axis and that the p38 MAPK pathway mediates the phosphorylation of CREB. Knockdown of CREB prevents chordin expression and mesoderm dorsalization by the Spemann organizer, whereas ectopic expression of activated CREB-VP16 chimera induces chordin expression and dorsalizes mesoderm. Expression of high levels of p38 activator, MKK6E or CREB-VP16 in embryos converts ventral mesoderm into a dorsal organizing center. p38 MAPK and CREB function downstream of maternal Wnt/β-catenin and the organizer-specific genes siamois and goosecoid. At low expression levels, MKK6E induces expression of lateral genes without inducing the expression of dorsal genes. Loss of CREB or p38 MAPK activity enables the expansion of the ventral homeobox gene vent1 into the dorsal marginal region, preventing the lateral expression of Xmyf5. Overall, these data indicate that dorsal-ventral mesoderm patterning is regulated by differential p38/CREB activities along the axis.     

Reciprocal changes in calcification of the gastrolith and cuticle during the molt cycle of the red claw crayfish Cherax quadricarinatus

Mobilization of calcium during the molt cycle from the cuticle to transient calcium deposits is widely spread in crustaceans. The dynamics of calcium transport to transient calcium deposits called gastroliths and to the cuticle over the course of the molt cycle were studied in the crayfish Cherax quadricarinatus. In this species, calcium was deposited in the gastroliths during premolt and transported back to the cuticle during postmolt, shown by digital X-ray radiograph analysis. The predominant mineral in the crayfish is amorphous calcium carbonate embedded in an organic matrix composed mainly of chitin. Scanning electron micrographs of the cuticle during premolt showed that the endocuticle and parts of the exocuticle were the source of most of the labile calcium, while the epicuticle did not undergo degradation and remained mineralized throughout the molt cycle. The gastroliths are made of concentric layers of amorphous calcium carbonate intercalated between chitinous lamella. Measurements of pH and calcium levels during gastrolith deposition showed that calcium concentrations in the gastroliths, stomach, and muscle were about the same (10 to 11 mmol l(-1)). On the other hand, pH varied greatly, from 8.7+/-0.15 in the gastrolith cavity through 7.6+/-0.2 in muscle to 6.9+/-0.5 in the stomach.     

Effects of acupuncture on rates of pregnancy and live birth among women undergoing in vitro fertilisation: systematic review and meta-analysis

Objective To evaluate whether acupuncture improves rates of pregnancy and live birth when used as an adjuvant treatment to embryo transfer in women undergoing in vitro fertilisation.Design Systematic review and meta-analysis.Data sources Medline, Cochrane Central, Embase, Chinese Biomedical Database, hand searched abstracts, and reference lists.Review methods Eligible studies were randomised controlled trials that compared needle acupuncture administered within one day of embryo transfer with sham acupuncture or no adjuvant treatment, with reported outcomes of at least one of clinical pregnancy, ongoing pregnancy, or live birth. Two reviewers independently agreed on eligibility; assessed methodological quality; and extracted outcome data. For all trials, investigators contributed additional data not included in the original publication (such as live births). Meta-analyses included all randomised patients.Data synthesis Seven trials with 1366 women undergoing in vitro fertilisation were included in the meta-analyses. There was little clinical heterogeneity. Trials with sham acupuncture and no adjuvant treatment as controls were pooled for the primary analysis. Complementing the embryo transfer process with acupuncture was associated with significant and clinically relevant improvements in clinical pregnancy (odds ratio 1.65, 95% confidence interval 1.27 to 2.14; number needed to treat (NNT) 10 (7 to 17); seven trials), ongoing pregnancy (1.87, 1.40 to 2.49; NNT 9 (6 to 15); five trials), and live birth (1.91, 1.39 to 2.64; NNT 9 (6 to 17); four trials). Because we were unable to obtain outcome data on live births for three of the included trials, the pooled odds ratio for clinical pregnancy more accurately represents the true combined effect from these trials rather than the odds ratio for live birth. The results were robust to sensitivity analyses on study validity variables. A prespecified subgroup analysis restricted to the three trials with the higher rates of clinical pregnancy in the control group, however, suggested a smaller non-significant benefit of acupuncture (odds ratio 1.24, 0.86 to 1.77).Conclusions Current preliminary evidence suggests that acupuncture given with embryo transfer improves rates of pregnancy and live birth among women undergoing in vitro fertilisation.     

Peptide inhibitors of HIV-1 integrase: From mechanistic studies to improved lead compounds

The HIV-1 integrase enzyme (IN) catalyzes integration of viral DNA into the host genome. We previously developed peptides that inhibit IN in vitro and HIV-1 replication in cells. Here we present the design, synthesis and evaluation of several derivatives of one of these inhibitory peptides, the 20-mer IN1. The peptide corresponding to the N-terminal half of IN1 (IN1 1–10) was easier to synthesize and much more soluble than the 20-mer IN1. IN1 1–10 bound IN with improved affinity and inhibited IN activity as well as HIV replication and integration in infected cells. While IN1 bound the IN tetramer, its shorter derivatives bound dimeric IN. Mapping the peptide binding sites in IN provided a model that explains this difference. We conclude that IN1 1–10 is an improved lead compound for further development of IN inhibitors.