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"Dixon, David"
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A skateboard cat-astrophe
Everyone in Bayport is talking about the first annual skateboard contest. The winner gets free mac and cheese balls from The Easy Cheesy food truck for the whole year! Frank and Joe just wish there was someone to beat the school's biggest bully, and best skateboarder, Diesel Diffendorfer. Then their best friend Chet shows them a video of Skeeter the skateboarding cat, a cat that happens to belong to the new kid in town, Carlos Martinez. Suddenly, a plan is hatched.
Book
Single-cell RNA-seq reveals new types of human blood dendritic cells, monocytes, and progenitors
Blood contains many types of cells, including many immune system components. Immune cells used to be characterized by marker-based assays, but now classification relies on the genes that cells express. Villani et al. used deep sequencing at the single-cell level and unbiased clustering to define six dendritic cell and four monocyte populations. This refined analysis has identified, among others, a previously unknown dendritic cell population that potently activates T cells. Further cell culture revealed possible differentiation progenitors within the different cell populations. Science , this issue p. eaah4573 Discovery of additional immune cell subtypes will help identify functions and immune monitoring during disease. Dendritic cells (DCs) and monocytes play a central role in pathogen sensing, phagocytosis, and antigen presentation and consist of multiple specialized subtypes. However, their identities and interrelationships are not fully understood. Using unbiased single-cell RNA sequencing (RNA-seq) of ~2400 cells, we identified six human DCs and four monocyte subtypes in human blood. Our study reveals a new DC subset that shares properties with plasmacytoid DCs (pDCs) but potently activates T cells, thus redefining pDCs; a new subdivision within the CD1C + subset of DCs; the relationship between blastic plasmacytoid DC neoplasia cells and healthy DCs; and circulating progenitor of conventional DCs (cDCs). Our revised taxonomy will enable more accurate functional and developmental analyses as well as immune monitoring in health and disease.
Journal Article
Enzyme activities and subcellular localization of members of the Arabidopsis glutathione transferase superfamily
Enzyme screens with Strep-tagged recombinant proteins and expression studies with the respective green fluorescent protein (GFP) fusions have been employed to examine the functional activities and subcellular localization of members of the Arabidopsis glutathione transferase (GST) superfamily. Fifty-one of 54 GST family members were transcribed and 41 found to express as functional glutathione-dependent enzymes in Escherichia coli. Functional redundancy was observed and in particular three theta (T) class GSTs showed conserved activities as hydroperoxide-reducing glutathione peroxidases (GPOXs). When expressed in tobacco as GFP fusions, all three GSTTs localized to the peroxisome, where their GPOX activity could prevent membrane damage arising from fatty acid oxidation. Through alternative splicing, two of these GSTTs form fusions with Myb transcription factor-like domains. Examination of one of these variants showed discrete localization within the nucleus, possibly serving a role in reducing nucleic acid hydroperoxides or in signalling. Based on this unexpected differential sub-cellular localization, 15 other GST family members were expressed as GFP fusions in tobacco. Most accumulated in the cytosol, but GSTU12 localized to the nucleus, a family member resembling a bacterial tetrachlorohydroquinone dehalogenase selectively associated with the plasma membrane, and a lambda GSTL2 was partially directed to the peroxisome after removal of a putative chloroplast transit peptide. Based on the results obtained with the GSTTs, it was concluded that these proteins can exert identical protective functions in differing subcellular compartments.
Journal Article
The hobbit : or There and back again
\"When Thorin Oakenshield and his band of dwarves embark upon a dangerous quest to reclaim the hoard of gold stolen from them by the evil dragon Smaug, Gandalf the wizard suggests an unlikely accomplice: Bilbo Baggins, an unassuming Hobbit dwelling in peaceful Hobbiton. Along the way, the company faces trolls, goblins, giant spiders, and worse. But as they journey from the wonders of Rivendell to the terrors of Mirkwood and beyond, Bilbo will find that there is more to him than anyone--himself included--ever dreamed. Unexpected qualities of courage and cunning, and a love of adventure, propel Bilbo toward his great destiny . . . a destiny that waits in the dark caverns beneath the Misty Mountains, where a twisted creature known as Gollum jealously guards a precious magic ring.\"-- Provided by publisher.
BOOK
Duration of protection from live attenuated vs. sub unit HSV-2 vaccines in the guinea pig model of genital herpes: Reassessing efficacy using endpoints from clinical trials
Although herpes simplex viruses (HSV) are a major target for vaccine development no vaccine is currently licensed.
A live attenuated HSV virus vaccine, VC2 was compared to a subunit HSV vaccine, glycoprotein D (gD2) administered with the adjuvant, MPL/Alum using the guinea pig model of genital herpes. Three doses of intramuscular (IM) vaccine were provided followed by intravaginal challenge with HSV-2 at either 3 weeks or six months after the last vaccination.
Both VC2 and gD2 vaccines reduced acute genital disease. VC2 was somewhat more effective in reducing acute vaginal replication, the amount of virus in neural tissue, subsequent recurrent disease and recurrent virus shedding following challenge at 3 weeks post vaccination. Both vaccines continued to provide protection at 6 months after vaccination but the differences between the vaccines became more pronounced in favor of the live attenuated vaccine, VC2. Significant differences in acute disease, acute vaginal virus replication, recurrent disease and recurrent virus shedding (P<0.05 for each) was observed comparing the vaccines. Re-examination of protection for this study using criteria similar to those used in recent clinical trials (inclusion of recurrent disease) showed that efficacy may not be as high in this model as previously thought prompting a need to assess the best predictive outcomes for protection in humans.
While both the live attenuated vaccine, VC2, and the gD2 subunit vaccine provided protection, the duration of protection appeared to be greater for VC2. Using the same evaluation criteria as used in human trials provided unique insights into the utility of the guinea pig model.
Journal Article
The Hobbit : or there and back again
Bilbo Baggins, a quiet and contented hobbit, embarks on a grand and dangerous adventure when he joins the wizard Gandalf and a group of dwarves on their quest to reclaim a stolen treasure. Presented in graphic novel form.
BOOK
Substrate specificity and safener inducibility of the plant UDP‐glucose‐dependent family 1 glycosyltransferase super‐family
Summary Plants contain large numbers of family 1 UDP‐glucose‐dependent glycosyltransferases (UGTs), including members that conjugate xenobiotics. Arabidopsis contains 107 UGT genes with 99 family members successfully expressed as glutathione transferase (GST)‐fusion proteins in E. coli. A high‐throughput catalytic screen was developed based on quantification of the fusion by measuring GST activity. UGT activity using UDP‐glucose as donor was then determined using 11 synthetic acceptors bearing hydroxyl, amino and thiol groups that had been shown to undergo conjugation in plant extracts. In total, 44 UGTs, largely members of the D and E groups, were active towards xenobiotics, glucosylating phenol and thiol acceptors. In contrast, N‐glucosyltransferase (NGT) activity was almost exclusively restricted to a single enzyme, UGT72B1. Using DNA microarrays, the induction of UGT transcripts following treatment with the herbicide safener fenclorim was compared in Arabidopsis and rice. D and L group members were the most safener‐inducible UGTs in both species. The respective Arabidopsis enzymes showed low conjugating activity towards xenobiotics. Using Genevestigator, a small group of safened D and L UGTs were consistently induced in response to biotic and abiotic stress suggestive of protective activities beyond xenobiotic detoxification in both species. The induction of other detoxifying gene families following treatment with fenclorim, namely cytochromes P450 and glutathione transferases, further confirmed the selective enhancement of related subfamily members in the two species giving new insight into the safening response in cereals, where herbicide tolerance is enhanced compared with dicots, which are unresponsive to these treatments.
Journal Article
SpongeBob comics. #2, Aquatic adventurers, unite!
A collection of superhero-themed SpongeBob SquarePants comics includes the reunion of the Aquatic Adventurers and SpongeBob's turn as the Super Bubble Sponge.
Book
Protein-Ligand Fishing in planta for Biologically Active Natural Products Using Glutathione Transferases
Screening for natural products which bind to proteins
has been used to identify ligands of the plant-specific glutathione transferase (GST) tau (U) and phi (F) classes, that are present in large gene families in crops and weeds, but have largely undefined functions. When expressed as recombinant proteins in
these proteins have been found to tightly bind a diverse range of natural product ligands, with fatty acid-and porphyrinogen-derivatives associated with GSTUs and a range of heterocyclic compounds with GSTFs. With an interest in detecting the natural binding partners of these proteins
, we have expressed the two best characterized GSTs from
(
),
GSTF2 and
GSTU19, as
-tagged fusion proteins
Following transient and stable expression in Nicotiana and Arabidopsis, respectively, the GSTs were recovered using Strep-Tactin affinity chromatography and the bound ligands desorbed and characterized by LC-MS.
GSTF2 predominantly bound phenolic derivatives including
-glutathionylated lignanamides and methylated variants of the flavonols kaempferol and quercetin.
GSTU19 captured glutathionylated conjugates of oxylipins, indoles, and lignanamides. Whereas the flavonols and oxylipins appeared to be authentic
ligands, the glutathione conjugates of the lignanamides and indoles were artifacts formed during extraction. When tested for their binding characteristics, the previously undescribed indole conjugates were found to be particularly potent inhibitors of
GSTU19. Such ligand fishing has the potential to both give new insight into protein function
as well as identifying novel classes of natural product inhibitors of enzymes of biotechnological interest such as GSTs.
Journal Article