Rapidly evolving RNA viruses, such as the GII.4 strain of human norovirus (HuNoV), and their vaccines elicit complex serological responses associated with previous exposure. Specific correlates of ...protection, moreover, remain poorly understood. Here, we report the GII.4-serological antibody repertoire—pre- and post-vaccination—and select several antibody clonotypes for epitope and structural analysis. The humoral response was dominated by GII.4-specific antibodies that blocked ancestral strains or by antibodies that bound to divergent genotypes and did not block viral-entry-ligand interactions. However, one antibody, A1431, showed broad blockade toward tested GII.4 strains and neutralized the pandemic GII.P16-GII.4 Sydney strain. Structural mapping revealed conserved epitopes, which were occluded on the virion or partially exposed, allowing for broad blockade with neutralizing activity. Overall, our results provide high-resolution molecular information on humoral immune responses after HuNoV vaccination and demonstrate that infection-derived and vaccine-elicited antibodies can exhibit broad blockade and neutralization against this prevalent human pathogen.
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•Serum vaccine response is dominated by a small number of abundant antibody clonotypes•Vaccine-boosted antibodies predominantly target conserved norovirus epitopes•Identified cross-genogroup and strain-specific epitopes•Discovered a pandemic-genotype neutralizing antibody recognizing a conserved epitope
Human norovirus (HuNoV) is a leading cause of gastroenteritis. Lindesmith et al. identify circulating serum antibodies following experimental HuNoV vaccination in humans and map them to viral epitopes. One antibody recognizes a neutralizing epitope conserved across three decades of pandemic strains and neutralizes virus in vitro, demonstrating that vaccination can elicit pandemic-strain neutralizing antibody responses in some individuals.
Flow chemistry was initially used for speed to early phase material delivery in the development laboratories, scaling up chemical transformations that we would not or could not scale up batch for ...safety reasons. Some early examples included a Newman Kwart Rearrangement, Claisen rearrangement, hydroformylation, and thermal imidazole cyclization. Next, flow chemistry was used to enable safe scale up of hazardous chemistries to manufacturing plants. Examples included high pressure hydrogenation, aerobic oxidation, and Grignard formation reactions. More recently, flow chemistry was used in Small Volume Continuous (SVC) processes, where highly potent oncolytic molecules were produced by fully continuous processes at about 10 kg/day including reaction, extraction, distillation, and crystallization, using disposable equipment contained in fume hoods.
The ketone intermediate LSN647712 is a key synthetic intermediate for the drug substance lasmiditan manufacturing process. A three-step connected continuous flow process utilizing a Turbo Grignard ...reagent, N-methylpiperidin-4-ylmagnesium chloride, and lithiated 2,6-dibromopyridine sequentially added to double electrophile (OC(++) synthon dimethylcarbamyl chloride (DMCC) was developed to deliver the ketone intermediate in a high chemical yield (>85%). This highly productive (>100 g/h lab system) and intensified process (τ ∼ 3 min) yields the product in high purity upon batch reactive crystallization to form a corresponding hydrobromide salt. In addition to the connected plug flow reactor system, the Grignard reagent, N-methylpiperidin-4-ylmagnesium chloride, was also prepared continuously in CSTR as a more soluble LiCl adduct in THF (Turbo Grignard).
A single, recessive mutation in soybean (Glycine max L. Merr.), which confers a seed phenotype of increased inorganic phosphate, decreased phytic acid, and a decrease in total raffinosaccharides, has ...been previously disclosed (S.A. Sebastian, P.S. Kerr, R.W. Pearlstein, W.D. Hitz 2000 Soy in Animal Nutrition, pp 56-74). The genetic lesion causing the multiple changes in seed phenotype is a single base change in the third base of the codon for what is amino acid residue 396 of the mature peptide encoding a seed-expressed myo-inositol 1-phospate synthase gene. The base change causes residue 396 to change from lysine to asparagine. That amino acid change decreases the specific activity of the seed-expressed myo-inositol 1-phosphate synthase by about 90%. Radio tracer experiments indicate that the supply of myo-inositol to the reaction, which converts UDP-galactose and myo-inositol to galactinol is a controlling factor in the conversion of total carbohydrate into the raffinosaccharides in both wild-type and mutant lines. That same decrease in myo-inositol 1-phosphate synthetic capacity leads to a decreased capacity for the synthesis of myo-inositol hexaphosphate (phytic acid) and a concomitant increase in inorganic phosphate.
Diet influences host metabolism and intestinal microbiota; however, detailed understanding of this tripartite interaction is limited. To determine whether the nonfermentable fiber hydroxypropyl ...methylcellulose (HPMC) could alter the intestinal microbiota and whether such changes correlated with metabolic improvements, C57B/L6 mice were normalized to a high‐fat diet (HFD), then either maintained on HFD (control), or switched to HFD supplemented with 10% HPMC, or a low‐fat diet (LFD). Compared to control treatment, both LFD and HPMC reduced weight gain (11.8 and 5.7 g, respectively), plasma cholesterol (23.1 and 19.6%), and liver triglycerides (73.1 and 44.6%), and, as revealed by 454‐pyrosequencing of the microbial 16S rRNA gene, decreased microbial α‐diversity and differentially altered intestinal microbiota. Both LFD and HPMC increased intestinal Erysipelotrichaceae (7.3‐ and 12.4‐fold) and decreased Lachnospiraceae (2.0‐ and 2.7‐fold), while only HPMC increased Peptostreptococcaceae (3.4‐fold) and decreased Ruminococcaceae (2.7‐fold). Specific microorganisms were directly linked with weight change and metabolic parameters in HPMC and HFD mice, but not in LFD mice, indicating that the intestinal microbiota may play differing roles during the two dietary modulations. This work indicates that HPMC is a potential prebiotic fiber that influences intestinal microbiota and improves host metabolism.—Cox, L. M., Cho, I., Young, S. A., Kerr Anderson, W. H., Waters, B. J., Hung, S.‐C., Gao, Z., Mahana, D., Bihan, M., Alekseyenko, A. V., Methé, B. A., Blaser, M. J. The nonfermentable dietary fiber hydroxypropyl methylcellulose modulates intestinal microbiota. FASEB J. 27, 692–702 (2013). www.fasebj.org
Correlational evidence suggests that animals may use changes in barometric pressure to predict or respond to changes in weather. Birds adjust the timing of migratory flights and migratory ...restlessness in response to changing weather, and they make facultative movements in response to storms during winter and breeding. Using the pressure chamber of a hypobaric climatic wind tunnel we tested the responses of white-throated sparrows, Zonotrichia albicollis, to experimental changes in air pressure alone, or air pressure and temperature in combination. Sparrows in wintering (short-day) condition were exposed to gradual changes in pressure/temperature at dawn that simulated large but realistic high- and low-pressure weather systems. During a drop in pressure, birds approached their food cup more quickly and moved more often. There was no effect of increasing pressure and no additional effects of temperature change. Sparrows in spring migratory condition (photostimulated) were exposed to pressure/temperature changes in the evening. Decreases in temperature resulted in less migratory restlessness during the first hour of night, but there was no additional effect of pressure changes. These experimental results indicate that white-throated sparrows can facultatively adjust their behaviour in direct response to changing barometric pressure and temperature.
•Changes in the weather are associated with changes in barometric pressure and temperature.•We experimentally exposed sparrows to changes in air pressure and temperature.•Decreasing air pressure led to faster foraging at dawn under winter conditions.•Decreased temperature reduced spring migratory behaviour.
To explore factors associated with sugar‐sweetened beverage (SSB) consumption in Australia.
Pooled data from Western Australian (WA) and South Australian (SA) 2009 and 2012 nutrition monitoring ...survey series interviews of 2,832 WA and 10,764 SA adults aged 18 to 64 years. Demographic data were collected and independent samples t‐test, analysis of variance, multiple logistic regression performed.
Obese participants were more likely to consume SSB than healthy weight participants (SA: OR=1.77; 95% CI 1.56–2.02; WA: OR=1.53; 1.05–2.24). SA obese participants consumed more SSB per day (152.0 mL; 140.7–163.5) than healthy weight (80.1 mL; 73.2–88.2; p<0.001) and overweight participants (106.9 mL; 99.0, 114.8; p<0.001). Males were more likely to consume SSB than females (SA: OR 1.80; 1.35–2.40; WA: 1.81; 1.64–2.00). WA participants who didn’t think about the healthiness of food (4.55; 2.71–7.64) and bought meals away from home the day prior (1.55; 1.15–2.09) were more likely to consume SSB. SA adults rating their health highest were less likely to consume SSB (0.62; 0.54–0.72).
SSB consumers are more likely to be male, have little interest in health, or have purchased a meal away from home.
Increasing awareness of the adverse health effects of consumption may be a first step in curbing SSB intake.
The ongoing evolution of Ebolaviruses poses significant challenges to the development of immunodiagnostics for detecting emergent viral variants. There is a critical need for the discovery of ...monoclonal antibodies with distinct affinities and specificities for different Ebolaviruses. We developed an efficient technology for the rapid discovery of a plethora of antigen-specific monoclonal antibodies from immunized animals by mining the VH:VL paired antibody repertoire encoded by highly expanded B cells in the draining popliteal lymph node (PLN). This approach requires neither screening nor selection for antigen-binding. Specifically we show that mouse immunization with Ebola VLPs gives rise to a highly polarized antibody repertoire in CD138(+) antibody-secreting cells within the PLN. All highly expanded antibody clones (7/7 distinct clones/animal) were expressed recombinantly, and shown to recognize the VLPs used for immunization. Using this approach we obtained diverse panels of antibodies including: (i) antibodies with high affinity towards GP; (ii) antibodies which bound Ebola VLP Kissidougou-C15, the strain circulating in the recent West African outbreak; (iii) non-GP binding antibodies that recognize wild type Sudan or Bundibugyo viruses that have 39% and 37% sequence divergence from Ebola virus, respectively and (iv) antibodies to the Reston virus GP for which no antibodies have been reported.
Epithelial tubes are the infrastructure for organs and tissues, and tube morphogenesis requires precise orchestration of cell signaling, shape, migration, and adhesion. Follicle cells in the ...Drosophila ovary form a pair of epithelial tubes whose lumens act as molds for the eggshell respiratory filaments, or dorsal appendages (DAs). DA formation is a robust and accessible model for studying the patterning, formation, and expansion of epithelial tubes. Tramtrack69 (TTK69), a transcription factor that exhibits a variable embryonic DNA-binding preference, controls DA lumen volume and shape by promoting tube expansion; the tramtrack mutation twin peaks (ttktwk) reduces TTK69 levels late in oogenesis, inhibiting this expansion. Microarray analysis of wild-type and ttktwk ovaries, followed by in situ hybridization and RNAi of candidate genes, identified the Phospholipase B-like protein Lamina ancestor (LAMA), the scaffold protein Paxillin, the endocytotic regulator Shibire (Dynamin), and the homeodomain transcription factor Mirror, as TTK69 effectors of DA-tube expansion. These genes displayed enriched expression in DA-tube cells, except lama, which was expressed in all follicle cells. All four genes showed reduced expression in ttktwk mutants and exhibited RNAi phenotypes that were enhanced in a ttktwk/+ background, indicating ttktwk genetic interactions. Although previous studies show that Mirror patterns the follicular epithelium prior to DA tubulogenesis, we show that Mirror has an independent, novel role in tube expansion, involving positive regulation of Paxillin. Thus, characterization of ttktwk-differentially expressed genes expands the network of TTK69 effectors, identifies novel epithelial tube-expansion regulators, and significantly advances our understanding of this vital developmental process.
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•Microarray, in situ, and RNAi identify Tramtrack69 (TTK69) tube-expansion effectors.•modENCODE's TTK69 embryo binding data indicate a highly variable binding preference.•We develop a method for dual immunostaining:FISH in the Drosophila ovary.•Tramtrack genetically interacts with lamina ancestor, shibire, Paxillin, and mirror.•Mirror regulates Paxillin in tube cells independently of its known role in patterning.
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