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Differential toxicity and venom gland gene expression in Centruroides vittatus
Variation in venom toxicity and composition exists in many species. In this study, venom potency and venom gland gene expression was evaluated in Centruroides vittatus, size class I-II (immature) and size class IV (adults/penultimate instars) size classes. Venom toxicity was evaluated by probit analysis and returned ED50 values of 50.1 μg/g for class IV compared to 134.2 μg/g for class I-II 24 hours post injection, suggesting size class IV was 2.7 fold more potent. Next generation sequencing (NGS and qPCR were used to characterize venom gland gene expression. NGS data was assembled into 36,795 contigs, and annotated using BLASTx with UNIPROT. EdgeR analysis of the sequences showed statistically significant differential expression in transcripts associated with sodium and potassium channel modulation. Sodium channel modulator expression generally favored size class IV; in contrast, potassium channel modulators were favored in size class I-II expression. Real-time quantitative PCR of 14 venom toxin transcripts detected relative expression ratios that paralleled NGS data and identified potential family members or splice variants for several sodium channel modulators. Our data suggests ontogenetic differences in venom potency and venom related genes expression exist between size classes I-II and IV.
Journal Article
The Loud house. #1, There will be chaos
\"Learn from Lincoln Loud the ins and outs of living in a huge household with 10 sisters!\"--Back cover.
Book
The AP-1 transcription factor Batf controls T(H)17 differentiation
Activator protein 1 (AP-1, also known as JUN) transcription factors are dimers of JUN, FOS, MAF and activating transcription factor (ATF) family proteins characterized by basic region and leucine zipper domains. Many AP-1 proteins contain defined transcriptional activation domains, but BATF and the closely related BATF3 (refs 2, 3) contain only a basic region and leucine zipper, and are considered to be inhibitors of AP-1 activity. Here we show that Batf is required for the differentiation of IL17-producing T helper (T(H)17) cells. T(H)17 cells comprise a CD4(+) T-cell subset that coordinates inflammatory responses in host defence but is pathogenic in autoimmunity. Batf(-/-) mice have normal T(H)1 and T(H)2 differentiation, but show a defect in T(H)17 differentiation, and are resistant to experimental autoimmune encephalomyelitis. Batf(-/-) T cells fail to induce known factors required for T(H)17 differentiation, such as RORgamma t (encoded by Rorc) and the cytokine IL21 (refs 14-17). Neither the addition of IL21 nor the overexpression of RORgamma t fully restores IL17 production in Batf(-/-) T cells. The Il17 promoter is BATF-responsive, and after T(H)17 differentiation, BATF binds conserved intergenic elements in the Il17a-Il17f locus and to the Il17, Il21 and Il22 (ref. 18) promoters. These results demonstrate that the AP-1 protein BATF has a critical role in T(H)17 differentiation.
Journal Article
The emergence of computational thinking in national mathematics curricula: An Australian example
As we move further into the digital age, the acquisition of digital literacy (DL) and computational thinking (CT) skills is emerging internationally as an essential goal for students in contemporary school curricula. As the world becomes more uncertain and volatile due to impacts of artificial intelligence (AI), international unrest, climate change, global economic instability and unforeseen catastrophes such as the Coronavirus (Covid-19) pandemic, it is necessary to review, revise and refine school education curricula and policies. The issue of what is essential for students to learn, and how they learn it, is of growing importance to international organisations such as the Organisation for Economic Co-operation and Development (OECD) and the United Nations Educational, Scientific and Cultural Organisation (UNESCO) and is emerging as a significant driver for educational reform across the globe. The growing prominence of CT and DL skills across many industry sectors has prompted recent changes to international assessment frameworks such as the Programme for International Student Assessment (PISA) and the Trends in International Mathematics and Science Study (TIMSS). This paper will briefly discuss specific examples of alternative approaches to addressing CT in national curricula for the compulsory years of schooling and explain how CT has been adopted as a driver for mathematics curriculum change in Australia.
Journal Article
The Exosome Component Rrp6 Is Required for RNA Polymerase II Termination at Specific Targets of the Nrd1-Nab3 Pathway
The exosome and its nuclear specific subunit Rrp6 form a 3'-5' exonuclease complex that regulates diverse aspects of RNA biology including 3' end processing and degradation of a variety of noncoding RNAs (ncRNAs) and unstable transcripts. Known targets of the nuclear exosome include short (<1000 bp) RNAPII transcripts such as small noncoding RNAs (snRNAs), cryptic unstable transcripts (CUTs), and some stable unannotated transcripts (SUTs) that are terminated by an Nrd1, Nab3, and Sen1 (NNS) dependent mechanism. NNS-dependent termination is coupled to RNA 3' end processing and/or degradation by the Rrp6/exosome in yeast. Recent work suggests Nrd1 is necessary for transcriptome surveillance, regulating promoter directionality and suppressing antisense transcription independently of, or prior to, Rrp6 activity. It remains unclear whether Rrp6 is directly involved in termination; however, Rrp6 has been implicated in the 3' end processing and degradation of ncRNA transcripts including CUTs. To determine the role of Rrp6 in NNS termination globally, we performed RNA sequencing (RNA-Seq) on total RNA and perform ChIP-exo analysis of RNA Polymerase II (RNAPII) localization. Deletion of RRP6 promotes hyper-elongation of multiple NNS-dependent transcripts resulting from both improperly processed 3' RNA ends and faulty transcript termination at specific target genes. The defects in RNAPII termination cause transcriptome-wide changes in mRNA expression through transcription interference and/or antisense repression, similar to previously reported effects of depleting Nrd1 from the nucleus. Elongated transcripts were identified within all classes of known NNS targets with the largest changes in transcription termination occurring at CUTs. Interestingly, the extended transcripts that we have detected in our studies show remarkable similarity to Nrd1-unterminated transcripts at many locations, suggesting that Rrp6 acts with the NNS complex globally to promote transcription termination in addition to 3' end RNA processing and/or degradation at specific targets.
Journal Article
RNA Polymerase II CTD phosphatase Rtr1 fine-tunes transcription termination
RNA Polymerase II (RNAPII) transcription termination is regulated by the phosphorylation status of the C-terminal domain (CTD). The phosphatase Rtr1 has been shown to regulate serine 5 phosphorylation on the CTD; however, its role in the regulation of RNAPII termination has not been explored. As a consequence of RTR1 deletion, interactions within the termination machinery and between the termination machinery and RNAPII were altered as quantified by Disruption-Compensation (DisCo) network analysis. Of note, interactions between RNAPII and the cleavage factor IA (CF1A) subunit Pcf11 were reduced in rtr1Δ, whereas interactions with the CTD and RNA-binding termination factor Nrd1 were increased. Globally, rtr1Δ leads to decreases in numerous noncoding RNAs that are linked to the Nrd1, Nab3 and Sen1 (NNS) -dependent RNAPII termination pathway. Genome-wide analysis of RNAPII and Nrd1 occupancy suggests that loss of RTR1 leads to increased termination at noncoding genes. Additionally, premature RNAPII termination increases globally at protein-coding genes with a decrease in RNAPII occupancy occurring just after the peak of Nrd1 recruitment during early elongation. The effects of rtr1Δ on RNA expression levels were lost following deletion of the exosome subunit Rrp6, which works with the NNS complex to rapidly degrade a number of noncoding RNAs following termination. Overall, these data suggest that Rtr1 restricts the NNS-dependent termination pathway in WT cells to prevent premature termination of mRNAs and ncRNAs. Rtr1 facilitates low-level elongation of noncoding transcripts that impact RNAPII interference thereby shaping the transcriptome.
Journal Article
Newly modified ‘pseudo flap’ without compromising vascularity to enhance repair of long distal ureteral loss: A retrospective analysis of a prospective database
Objective To present an alternative technique called pseudo‐flap for reconstructing long ureteral defects as an alternative to Boari flap. Despite being used for more than 70 years by urologists for tension‐free reconstruction of distal and mid‐ureteral defects, the Boari flap exhibits high complication rates, with an average of 27% (range 5.5%–30.4%). These complications arise from compromised blood supply, attributed to incisions made on all three sides of the flap and dependence on the flap base as the sole source of blood supply. Methods We retrospectively reviewed patients who underwent our modified technique by a single surgeon between 2008 and 2021. We used a semi‐oblique cystotomy on the lowest part of the anterior and contralateral aspects of the bladder after complete release from adhesions and sacrificing the superior vesical pedicle, if necessary. The innovative part of the technique involved making short relaxing incisions at different levels on both sides of a pseudo‐flap while pushing the bladder dome upward to reach the healthy ureter in a tension‐free manner, followed by anastomosis with a non‐refluxing or refluxing technique. Results Fifteen patients underwent the pseudo‐flap technique with a mean follow‐up of 16.9 months. Four had prior radiation, three had hyperthermic intraperitoneal chemotherapy (HIPEC) for peritoneal carcinomatosis, and one had a ureteral stricture in a transplanted kidney. Eight procedures were performed during intraoperative consultations. Only one patient (7%) developed a major complication (Clavien–Dindo grade ≥2). This patient developed postoperative leak, and none developed obstructive hydronephrosis, suggesting stricture or flap ischemia. The mean length of the flap was 9.3 cm. Conclusion Our pseudo‐flap technique has lower complication rates than the traditional Boari flap. It is not technically challenging, minimally compromises blood supply and is thus especially suitable for complex, highly morbid patients with decreased tissue vascularity, such as those with prior radiation and peritoneal carcinomatosis.
Journal Article
Learning speech-internal cues to pronoun interpretation from co-speech gesture: a training study
This study explores whether children can learn a structural processing bias relevant to pronoun interpretation from brief training. Over three days, 42 five-year-olds were exposed to narratives exhibiting a first-mentioned tendency. Two characters were introduced, and the first-mentioned was later described engaging in a solo activity. In our primary condition of interest, the Gesture Training condition, the solo-activity sentence contained an ambiguous pronoun, but co-speech gesture clarified the referent. There were two comparison conditions. In the Gender Training condition the characters were different genders, thereby avoiding ambiguity. In the Name Training condition, the first-mentioned name was simply repeated. Ambiguous pronoun interpretation was tested pre- and post-training. Children in the Gesture condition were significantly more likely to interpret ambiguous pronouns as the first-mentioned character after training. Results from the comparison conditions were ambiguous: there was a small but non-significant effect of training, but also no significant differences between conditions.
Journal Article