Tryptophan synthase uses an atypical mechanism to achieve substrate specificity

Tryptophan synthase (TrpS) catalyzes the final steps in the biosynthesis of L-tryptophan from L-serine (Ser) and indole-3-glycerol phosphate (IGP). We report that native TrpS can also catalyze a productive reaction with L-threonine (Thr), leading to (2S,3S)-β-methyltryptophan. Surprisingly, β-substitution occurs in vitro with a 3.4-fold higher catalytic efficiency for Ser over Thr using saturating indole, despite >82,000-fold preference for Ser in direct competition using IGP. Structural data identify a novel product binding site and kinetic experiments clarify the atypical mechanism of specificity: Thr binds efficiently but decreases the affinity for indole and disrupts the allosteric signaling that regulates the catalytic cycle

HENNOVATION: Learnings from Promoting Practice-Led Multi-Actor Innovation Networks to Address Complex Animal Welfare Challenges within the Laying Hen Industry

The Hennovation project, an EU H2020 funded thematic network, aimed to explore the potential value of practice-led multi-actor innovation networks within the laying hen industry. The project proposed that husbandry solutions can be practice-led and effectively supported to achieve durable gains in sustainability and animal welfare. It encouraged a move away from the traditional model of science providing solutions for practice, towards a collaborative approach where expertise from science and practice were equally valued. During the 32-month project, the team facilitated 19 multi-actor networks in five countries through six critical steps in the innovation process: problem identification, generation of ideas, planning, small scale trials, implementation and sharing with others. The networks included farmers, processors, veterinarians, technical advisors, market representatives and scientists. The interaction between the farmers and the other network actors, including scientists, was essential for farmer innovation. New relationships emerged between the scientists and farmers, based on experimental learning and the co-production of knowledge for improving laying hen welfare. The project demonstrated that a practice-led approach can be a major stimulus for innovation with several networks generating novel ideas and testing them in their commercial context. The Hennovation innovation networks not only contributed to bridging the science-practice gap by application of existing scientific solutions in practice but more so by jointly finding new solutions. Successful multi-actor, practice-led innovation networks appeared to depend upon the following key factors: active participation from relevant actors, professional facilitation, moderate resource support and access to relevant expertise. Farmers and processors involved in the project were often very enthusiastic about the approach, committing significant time to the network’s activities. It is suggested that the agricultural research community and funding agencies should place greater value on practice-led multi-actor innovation networks alongside technology and advisor focused initiatives to improve animal welfare and embed best practices

Phylloquinone (vitamin K 1 ) biosynthesis in plants: two peroxisomal thioesterases of lactobacillales origin hydrolyze 1,4‐dihydroxy‐2‐naphthoyl‐coa

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/1/TPJ_4972_sm_FigS3.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/2/TPJ_4972_sm_TableS1.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/3/TPJ_4972_sm_FigS2.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/4/TPJ_4972_sm_TableS4.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/5/TPJ_4972_sm_FigS6.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/6/j.1365-313X.2012.04972.x.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/7/TPJ_4972_sm_FigS1.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/8/TPJ_4972_sm_TableS3.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/9/TPJ_4972_sm_FigS5.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/10/TPJ_4972_sm_TableS2.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/92396/11/TPJ_4972_sm_FigS4.pd

Analysis of detector performance in a gigahertz clock rate quantum key distribution system

We present a detailed analysis of a gigahertz clock rate environmentally robust phase-encoded quantum key distribution (QKD) system utilizing several different single-photon detectors, including the first implementation of an experimental resonant cavity thin-junction silicon single-photon avalanche diode. The system operates at a wavelength of 850 nm using standard telecommunications optical fibre. A general-purpose theoretical model for the performance of QKD systems is presented with reference to these experimental results before predictions are made about realistic detector developments in this system. We discuss, with reference to the theoretical model, how detector operating parameters can be further optimized to maximize key exchange rates

The first Neanderthal remains from an open-air Middle Palaeolithic site in the Levant

The late Middle Palaeolithic (MP) settlement patterns in the Levant included the repeated use of caves and open landscape sites. The fossil record shows that two types of hominins occupied the region during this period - Neandertals and Homo sapiens. Until recently, diagnostic fossil remains were found only at cave sites. Because the two populations in this region left similar material cultural remains, it was impossible to attribute any open-air site to either species. In this study, we present newly discovered fossil remains from intact archaeological layers of the open-air site 'Ein Qashish, in northern Israel. The hominin remains represent three individuals: EQH1, a nondiagnostic skull fragment; EQH2, an upper right third molar (RM3); and EQH3, lower limb bones of a young Neandertal male. EQH2 and EQH3 constitute the first diagnostic anatomical remains of Neandertals at an open-air site in the Levant. The optically stimulated luminescence ages suggest that Neandertals repeatedly visited 'Ein Qashish between 70 and 60 ka. The discovery of Neandertals at open-air sites during the late MP reinforces the view that Neandertals were a resilient population in the Levant shortly before Upper Palaeolithic Homo sapiens populated the region

Management of Primocane-fruiting Blackberry: Impacts on Yield, Fruiting Season, and Cane Architecture

Primocane management systems were compared for ‘Prime-Jan’® and ‘Prime-Jim’®, primocane-fruiting blackberry (Rubus L. subgenus Rubus, Watson), grown in a field planting in Aurora, OR. Treatments studied were: 1) no manipulation of primocanes (untipped; no floricanes); 2) untipped primocanes growing in the presence of floricanes; 3) untipped primocanes grown with rowcover in late winter to early spring; and 4) primocanes “soft-tipped” at 1 m to encourage branching. Date of primocane first bloom and cane height at bloom were unaffected by cultivar and were only affected by primocane management in 2005. The number of growing degree-days to first bloom ranged from 1272 to 1390 depending on year. Primocane management did not affect ovule or drupelet number per berry or percent drupelet set. ‘Prime-Jim’ had more drupelets and greater weight per berry in 2005 than ‘Prime-Jan’. Fruit harvested earlier in the season had more ovules and drupelets than later harvested fruit in 2004. Primocanes that grew in the presence of floricanes were longer and bloomed later but did not differ in yield from untipped canes grown only for a primocane crop. Use of rowcover in 2005 advanced bloom and harvest, improving yield 73% compared with untipped control canes. Soft-tipping primocanes increased yield 114% to 150% compared with untipped canes (5.6 vs. 2.4 t·ha⁻¹) through increasing branch and node number per cane and percentage of fruiting nodes; soft-tipping did not delay harvest. Yield/cane was negatively correlated with the number of fruiting canes/plot but positively correlated with branches/cane, total branch and cane length, number of nodes and percent fruiting nodes, fruit/cane, and berry weight. The proportion of fruiting nodes was greater on branches than on the main cane illustrating the importance of managing this type of blackberry to increase branch number for high yield.Keywords: Summer pruning, Fruit size, ‘Prime-Jim’®, Rubus, Tipping, Yield components, ‘Prime-Jan’

High Efficiency Planar Geometry Germanium-on-silicon Single-photon Avalanche Diode Detectors

This paper presents the performance of 26 μm and 50 μm diameter planar Ge-on-Si single-photon avalanche diode (SPAD) detectors. The addition of germanium in these detectors extends the spectral range into the short-wave infrared (SWIR) region, beyond the capability of already well-established Si SPAD devices. There are several advantages for extending the spectral range into the SWIR region including: reduced eye-safety laser threshold, greater attainable ranges, and increased depth resolution in range finding applications, in addition to the enhanced capability to image through obscurants such as fog and smoke. The time correlated single-photon counting (TCSPC) technique has been utilized to observe record low dark count rates, below 100 kHz at a temperature of 125 K for up to a 6.6 % excess bias, for the 26 μm diameter devices. Under identical experimental conditions, in terms of excess bias and temperature, the 50 μm diameter device consistently demonstrates dark count rates a factor of 4 times greater than 26 μm diameter devices, indicating that the dark count rate is proportional to the device volume. Single-photon detection efficiencies of up to ~ 29 % were measured at a wavelength of 1310 nm at 125 K. Noise equivalent powers (NEP) down to 9.8 × 10-17 WHz-1/2 and jitters < 160 ps are obtainable, both significantly lower than previous 100 μm diameter planar geometry devices, demonstrating the potential of these devices for highly sensitive and high-speed imaging in the SWIR

Epithelial Immunization Induces Polyfunctional CD8+ T Cells and Optimal Mousepox Protection.

We assessed several routes of immunization with vaccinia virus (VACV) in protecting mice against ectromelia virus (ECTV). By a wide margin, skin scarification provided the greatest protection. Humoral immunity and resident-memory T cells notwithstanding, several approaches revealed that circulating, memory CD8(+) T cells primed via scarification were functionally superior and conferred enhanced virus control. Immunization via the epithelial route warrants further investigation, as it may also provide enhanced defense against other infectious agents

Ectopic Expression of Vaccinia Virus E3 and K3 Cannot Rescue Ectromelia Virus Replication in Rabbit RK13 Cells

Citation: Hand, E. S., Haller, S. L., Peng, C., Rothenburg, S., & Hersperger, A. R. (2015). Ectopic Expression of Vaccinia Virus E3 and K3 Cannot Rescue Ectromelia Virus Replication in Rabbit RK13 Cells. Plos One, 10(3), 15. doi:10.1371/journal.pone.0119189As a group, poxviruses have been shown to infect a wide variety of animal species. However, there is individual variability in the range of species able to be productively infected. In this study, we observed that ectromelia virus (ECTV) does not replicate efficiently in cultured rabbit RK13 cells. Conversely, vaccinia virus (VACV) replicates well in these cells. Upon infection of RK13 cells, the replication cycle of ECTV is abortive in nature, resulting in a greatly reduced ability to spread among cells in culture. We observed ample levels of early gene expression but reduced detection of virus factories and severely blunted production of enveloped virus at the cell surface. This work focused on two important host range genes, named E3L and K3L, in VACV. Both VACV and ECTV express a functional protein product from the E3L gene, but only VACV contains an intact K3L gene. To better understand the discrepancy in replication capacity of these viruses, we examined the ability of ECTV to replicate in wild-type RK13 cells compared to cells that constitutively express E3 and K3 from VACV. The role these proteins play in the ability of VACV to replicate in RK13 cells was also analyzed to determine their individual contribution to viral replication and PKR activation. Since E3L and K3L are two relevant host range genes, we hypothesized that expression of one or both of them may have a positive impact on the ability of ECTV to replicate in RK13 cells. Using various methods to assess virus growth, we did not detect any significant differences with respect to the replication of ECTV between wild-type RK13 compared to versions of this cell line that stably expressed VACV E3 alone or in combination with K3. Therefore, there remain unanswered questions related to the factors that limit the host range of ECTV