bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tyramine and itsAmtyr1receptor modulate attention in honey bees (Apis mellifera)
Authors: Latshaw Joseph S , Mazade Reece E , Petersen Mary , Mustard Julie A , Sinakevitch Irina , Wissler Lothar , Guo Xiaojiao , Cook Chelsea N. , Lei Hong , Gadau Jürgen , Smith Brian H 
Published: September 2022

AbstractAnimals must learn to ignore stimuli that are irrelevant to survival and attend to ones that enhance survival. When a stimulus regularly fails to be associated with an important consequence, subsequent excitatory learning about that stimulus can be delayed, which is a form of non-associative conditioning called ‘latent inhibition’. Honey bees show latent inhibition toward an odor they have experienced without association with food reinforcement. Moreover, individual honey bees from the same colony differ in the degree to which they show latent inhibition, and these individual differences have a genetic basis. To investigate the mechanisms that underly individual differences in latent inhibition, we selected two honey bee lines for high and low latent inhibition, respectively. We crossed those lines and mapped a Quantitative Trait Locus (QTL) for latent inhibition to a region of the genome that contains the tyramine receptor geneAmtyr11. We then show that disruption ofAmtyr1signaling either pharmacologically or through RNAi qualitatively changes the expression of latent inhibition but has little effect on appetitive conditioning, and these results suggest that AmTYR1 modulates inhibitory processing in the CNS. Electrophysiological recordings from the brain during pharmacological blockade are consistent with a model that AmTYR1 indirectly regulates Hebbian plasticity at inhibitory synapses in the CNS. Finally, sequencingAmtyr1and its up and downstream genomic region for high and low line workers suggested that individual differences might arise from variation in transcriptional control rather than structural changes in the coding region. Our results therefore identify a distinct modulatory pathway for this type of non-associative learning, and we propose a model for how it modulates Hebbian plasticity at defined synapses in the CNS. We have shown elsewhere how this modulation also underlies potentially adaptive intracolonial learning differences among individuals that benefit colony survival.
Zinc Fingers and Homeoboxes 2 is Required for Diethylnitrosamine-induced Liver Tumor Formation in C57BL/6 Mice
Authors: Jiang Jieyun , Turpin Courtney , Qiu Guofang (Shirley) , Xu Mei , Lee Eun , Hinds Terry D. , Peterson Martha L. , Spear Brett T. 
Published: September 2022

AbstractLiver cancer, comprised mainly of hepatocellular carcinoma (HCC), is the third leading cause of cancer deaths worldwide and increasing in Western countries. We previously identified the transcription factor Zinc Fingers and Homeoboxes 2 (Zhx2) as a regulator of hepatic gene expression, and many Zhx2 target genes are dysregulated in HCC. Here, we investigate HCC in Zhx2-deficient mice using the Diethylnitrosamine (DEN)-induced liver tumor model. Our study using whole-body Zhx2 knock-out (Zhx2KO) mice revealed the complete absence of liver tumors 9 and 10 months after DEN exposure.Analysis soon after DEN treatment showed no differences in expression of the DEN bioactivating enzyme CYP2E1 and DNA polymerase delta 2, or in the numbers of γH2AX foci between Zhx2KO and wild-type (Zhx2wt) mice. The absence of Zhx2, therefore, did not alter DEN bioactivation or DNA damage. Zhx2KO livers showed fewer positive foci for Ki67 staining and reduced IL-6 and AKT2 expression compared to Zhx2wt livers, suggesting that Zhx2 loss reduces liver cell proliferation and may account for reduced tumor formation. Tumors were reduced but not absent in DEN-treated liver-specific Zhx2 knock-out mice, suggesting that Zhx2 acts in both hepatocytes and non-parenchymal cells to inhibit tumor formation. Analysis of data from The Cancer Genome Atlas and Clinical Proteomic Tumor Consortium indicated that ZHX2 mRNA and protein levels were significantly higher in HCC patients and associated with clinical pathological parameters.ConclusionsIn contrast to previous studies in human hepatoma cell lines and other HCC mouse models showing that Zhx2 acts as a tumor suppressor, our data indicate that Zhx2 acts as an oncogene in the DEN-induced HCC model and is consistent with the higher ZHX2 expression in HCC patients.
Hsp90 provides a platform for kinase dephosphorylation by PP5
Authors: Jaime-Garza Maru , Nowotny Carlos , Coutandin Daniel , Wang Feng , Tabios Mariano , Agard David A. 
Published: September 2022

AbstractThe Hsp90 molecular chaperone collaborates with the phosphorylated Cdc37 cochaperone to maintain kinase proteostasis through the folding and activation of its many client kinases. As with many kinases, the Hsp90 client kinase CRaf is activated by phosphorylation at specific regulatory sites. The cochaperone phosphatase PP5 dephosphorylates CRaf but also Cdc37 in an Hsp90-dependent manner. Although dephosphorylating Cdc37 has been proposed as a mechanism for releasing Hsp90-bound kinases, here we show that Hsp90 bound kinases sterically inhibit Cdc37 dephosphorylation indicating kinase release must occur before Cdc37 dephosphorylation. The cryo-EM structure of PP5 in complex with Hsp90:Cdc37:CRaf reveals how Hsp90 both activates PP5 and scaffolds its association with the bound CRaf to dephosphorylate a site at the C-terminus of the kinase domain. Thus, we directly show how Hsp90’s role in maintaining protein homeostasis goes beyond folding and activation to include post translationally modifying its client kinases.
A Beefy-R culture medium: replacing albumin with rapeseed protein isolates
Authors: Stout Andrew J. , Rittenberg Miriam L. , Shub Michelle , Saad Michael K. , Mirliani Addison B. , Kaplan David L. 
Published: September 2022

AbstractThe development of cost-effective serum-free media is essential for the economic viability of cultured meat. A key challenge facing this goal is high-cost recombinant albumin that is necessary in some available serum-free media formulations. As such, there is substantial interest in finding albumin alternatives which are low-cost, effective, scalable, sustainable, and suitable for food applications. Recently, a serum-free medium termed Beefy-9 was developed for bovine satellite cells (BSCs), which relied on recombinant albumin as a key component to replace fetal bovine serum. Here we alter Beefy-9 by replacing albumin with rapeseed protein isolate, a bulk-protein solution obtained from agricultural waste-streams through simple isoelectric protein precipitation. This new medium, termed Beefy-R, improves BSC growth compared with Beefy-9 while maintaining cell phenotype and myogenicity. These results offer an effective, low-cost, and sustainable alternative to albumin for serum-free culture of muscle stem cells, thereby addressing a key hurdle facing cultured meat production.
Trade-offs constrain adaptive pathways to T6 survival
Authors: MacGillivray Kathryn A. , Ng Siu Lung , Wiesenfeld Sophia , Guest Randi L. , Jubery Tahrima , Silhavy Thomas J. , Ratcliff William C. , Hammer Brian K. 
Published: September 2022

AbstractMany microbial communities are characterized by intense competition for nutrients and space. One way for an organism to gain control of these resources is by eliminating nearby competitors. The Type VI Secretion System (T6) is a nano-harpoon used by many bacteria to inject toxins into neighboring cells. While much is understood about mechanisms of T6-mediated toxicity, little is known about the ways that competitors can defend themselves against this attack, especially in the absence of their own T6. Here we use directed evolution to examine the evolution of T6 resistance, subjecting eight replicate populations of Escherichia coli to T6 attack by Vibrio cholerae. Over ~500 generations of competition, the E. coli evolved to survive T6 attack an average of 27-fold better than their ancestor. Whole genome sequencing reveals extensive parallel evolution. In fact, we found only two pathways to increased T6 survival: apaH was mutated in six of the eight replicate populations, while the other two populations each had mutations in both yejM and yjeP. Synthetic reconstruction of individual and combined mutations demonstrate that yejM and yjeP are synergistic, with yejM requiring the mutation in yejP to provide a benefit. However, the mutations we identified are pleiotropic, reducing cellular growth rates, and increasing susceptibility to antibiotics and elevated pH. These trade-offs underlie the effectiveness of T6 as a bacterial weapon, and help us understand how the T6 shapes the evolution of bacterial interactions.SignificanceBacteria are the most abundant organisms on Earth and often live in dense, diverse communities, where they interact with each other. One of the most common interactions is antagonism. While most research has focused on diffusible toxins (e.g., antibiotics), bacteria have also evolved a contact-dependent nano-harpoon, the Type VI Secretion System (T6), to kill neighboring cells and compete for resources. While the co-evolutionary dynamics of antibiotic exposure is well understood, no prior work has examined how targets of T6 evolve resistance. Here, we use experimental evolution to observe how an Escherichia coli target evolves resistance to T6 when it is repeatedly competing with a Vibrio cholerae killer. After 30 rounds of competition, we identified mutations in three genes that improve E. coli survival, but found that these mutations come at a cost to other key fitness components. Our findings provide new insight into how contact-dependent antagonistic interaction drives evolution in a polymicrobial community.
Derivation of a minimal functional XIST by combining human and mouse interaction domains
Authors: Navarro-Cobos Maria Jose , Morales-Guzman Suria Itzel , Baldry Sarah E.L. , Brown Carolyn J. 
Published: September 2022

AbstractXIST is a 17-19 kb long non-coding RNA critical for X-chromosome inactivation. Tandem repeats within the RNA serve as functional domains involved in the cis-limited recruitment of heterochromatic changes and silencing. To explore the sufficiency of these domains while generating a functional miniXIST for targeted silencing approaches, we tested inducible constructs integrated into 8p in a male cell line. Previous results suggested silencing could be accomplished with a transgene comprised of the repeat A which is highly conserved and critical for silencing; the repeat F which overlaps regulatory elements and the repeat E region which contributes to XIST localization by binding proteins such as CIZ1. As PRC1 is recruited through HNRNPK binding of repeats B - C - D we included a second ‘miniXIST’ comprising AFE with the mouse PID, a 600-nucleotide region of repeat B and C. Silencing of nearby genes was possible with and without PID, however, silencing more distally required the addition of PID. The recruitment of heterochromatic marks, evaluated by IF combined with RNA FISH, revealed that the AFE domains were sufficient only for the recruitment of CIZ1. However, miniXIST transgene recruited all marks, albeit not to full XIST levels. The ability of the PID domain to facilitate silencing and heterochromatic marks recruitment was unexpected, and inhibition of PRC1 suggested that many of these are PRC1-independent. These results suggest that the addition of this small region allowed the partial recruitment of all the changes induced by a full XIST, demonstrating the feasibility of finding a minimal functional XIST.
Discovery of thyrocyte heterogeneity reveals an essential role of Notch signaling in thyroid function and mammalian homeostasis
Authors: Mosteiro Lluc , Thao Nguyen Thi Thu , Hankeova Simona , Reichelt Mike , Vandriel Shannon M. , Lai Zijuan , Choudhury Feroza K. , Sangaraju Dewakar , Kamath Binita M. , Scherl Alexis , Piskol Robert , Siebel Christian W. 
Published: September 2022

SUMMARYThe thyroid functions at the apex of a web of endocrine organs that control cell growth, differentiation and metabolic homeostasis. Thyroid dysregulation significantly impacts human health in myriad ways with thyroid diseases standing as the most common endocrine disorder. Despite the essential role of the thyroid in human health, a high-resolution view of the cellular composition as well as molecular mechanisms that govern function of this crucial organ have been lacking. Employing the first single-cell analyses of adult mouse thyroid, we here report the discovery of unexpected thyrocyte heterogeneity, specifically three distinct thyrocyte subtypes marked by different metabolic and Notch signaling patterns. Using a battery of pharmacologic and genetic methods, we find that selective inhibition of Notch ligands and receptors disrupts thyrocyte mitochondrial activity and ROS production, thus decreasing levels of circulating thyroid hormones, inducing hypothyroidism and disrupting whole-body thermoregulation. We find an enriched frequency of hypothyroidism in children with Alagille Syndrome, a genetic disorder marked by Notch loss-of-function mutations, suggesting that our Notch-thyroid mechanisms are relevant in humans and directly account for Alagille hypothyroidism. Overall, our work reveals that Notch, although classically described as a developmental pathway that determines cell fate, controls homeostasis and thermoregulation in the adult through a mitochondria-based mechanism in a subset of thyrocytes. Our fine-grained picture of the thyroid unveils a novel understanding of this key metabolic organ and provides clinically impactful insights into its pathological dysfunctions.
Utility of stromal lymphocytes in diagnosis and predicting upgrade of B3 breast lesions from core biopsies
Authors: Kader Tanjina , Hendry Shona , Provenzano Elena , Jayawardana Madawa W , Pang Jia-Min , Elder Kenneth , Byrne David J , Tjoeka Lauren , Frazer Helen ML , House Eloise , Jayasinghe Sureshni , Keane Holly , Murugasu Anand , Rajan Neeha , Miligy Islam M , Green Andrew R , Rakha Emad A , Fox Stephen B , Mann G. Bruce , Campbell Ian G , Gorringe Kylie L 
Published: September 2022

AbstractFor more than two decades attempts have been made to identify a subset of women diagnosed with lesions with uncertain malignant potential (B3 lesions) who could safely be observed rather than being treated with surgical excision and/or chemoprevention. Various histopathological, clinical and imaging parameters for risk recommendation have been evaluated, with little impact on clinical practice. The primary reason for surgery is to rule out an upgrade lesion to either ductal carcinomain situ(DCIS) or invasive breast cancer (IBC). While on average 30% of these patients are upgraded after diagnostic biopsy, a large number are over treated,making this an important harm of screening.Here we evaluated stromal lymphocytes from B3 biopsies (n=264) as a predictive biomarker for upgrade. A higher number of stromal lymphocytes were observed in upgraded B3 lesions than non-upgraded (p< 0.01, zero inflated binomial model) for both ductal and papillary lesions (n=174). This observation was validated in an independent cohort (p<0.001, p<0.05, zero binomial model, ductal and papillary lesions, respectively) (n=90). Our data suggested that the presence of ≥5% of lymphocytes in the surrounding specialised stroma of B3 lesions are predictive of B3 lesions being upgraded with a specificity of 93% and 87% in our discovery and validation cohorts, respectively. The area under the curve (AUC) for the discovery cohort using lymphocyte count and age as variables was 0.77 and was validated with an AUC of 0.81 in the validation cohort.In conclusion, we can identify a subset of the patients at risk of upgrade with high specificity. Assessing the tumour microenvironment including stromal lymphocytes may contribute to reducing unnecessary surgeries in the clinic.
Detection and Role of Feline Apolipoprotein B mRNA-editing Enzyme Catalytic Polypeptide Subunit 3G-Like Protein in Feline Cells and Tissues
Authors: Assogba Barnabe D. , Chaudhary Shannon , Kaur Harmanpreet , Soo Raymond , Dela Cruz Mark A. P. , Assogba Jovy M. G. 
Published: September 2022

AbstractThe innate host defence system is designed to resist pathogenic microorganism infections. Despite the compelling scientific evidence, our understanding of the full potential of the mechanism is still unclear due to the complex interactions between hosts and invaders. We previously reported latency in cat mucosal infected with low-dose cell-associated feline immunodeficiency virus (102 and 103 infected cells). Here we investigated the expression of Apolipoprotein B mRNA-editing enzyme catalytic subunit 3G (APOBEC3G or A3G) in feline cells and tissues and whether its presence antagonizes the viral pre-integration complex resulting in partial or complete FIV latency. Total RNA and protein lysates were collected from cell lines, blood, and tissue samples. Real-time RT-PCR and western blot assays were used to quantify fA3G-like protein in cats exposed to high versus low-dose cell-associated FIV. We consistently detected fA3G-like protein in mock T-cell lines (E-CD4+, MYA-1, Crandell feline kidney cells) and primary bone marrow-derived macrophages with variable expressions in feline peripheral blood mononuclear cells (PBMC). In addition, the fA3G-like protein was found to interact with FIV group-specific antigen (Gag) protein through immunoprecipitation assays. The protein expression was utterly abrogated following FIV infection. However, in lytic FIV infection (in vivo), fA3G-like protein decreased in early post-infection, whereas latently infected cats showed stable expression. These data are the first report of the fA3G-like protein expression in felines and its abrogation in lytic but not in latent FIV-infected individuals. These results might provide new insight into the role of fA3G-like protein in the host defence mechanism against retrovirus infections.
Microphysiological vascular malformation model reveals a role of dysregulated Rac1 and mTORC1/2 in lesion formation
Authors: Aw Wen Yih , Cho Crescentia , Wang Hao , Cooper Anne Hope , Doherty Elizabeth L. , Rocco David , Huang Stephanie A. , Kubik Sarah , Whitworth Chloe P. , Armstrong Ryan , Hickey Anthony J. , Griffith Boyce , Kutys Matthew L. , Blatt Julie , Polacheck William J. 
Published: September 2022

AbstractSomatic activating mutations ofPIK3CAare associated with the development of vascular malformations (VMs). Here, we describe a microfluidic model ofPIK3CA-driven VMs consisting of human umbilical vein endothelial cells (HUVECs) expressingPIK3CAactivating mutations embedded in 3D hydrogels. We observed enlarged and irregular vessel phenotypes, consistent with clinical signatures and concomitant with PI3K-driven upregulation of Rac1/PAK, MEK/ERK, and mTORC1/2 signaling. We observed differential effects between Alpelisib, a PIK3CA inhibitor, and Rapamycin, an mTORC1 inhibitor, in mitigating matrix degradation and vascular network topology. While both drugs are effective in preventing vessel enlargement, Alpelisib suppressed mTORC2-dependent AKT1 phosphorylation and MEK/ERK signaling. Rapamycin failed to reduce MEK/ERK and mTORC2 activity and resulted in vascular hyperbranching, while inhibiting PAK, MEK1/2, and mTORC1/2 signaling mitigates abnormal growth and vascular dilation. Collectively, these findings establish anin vitroplatform for modeling VMs and confirm a role of dysregulated Rac1/PAK and mTORC1/2 signaling inPIK3CA-driven VMs.
Aquaporin regulates cell rounding through vacuole formation during endothelial-to-hematopoietic transition
Authors: Sato Yuki , Shigematsu Mugiho , Shibata-Kanno Maria , Maejima Sho , Tamura Chie , Sakamoto Hirotaka 
Published: September 2022

ABSTRACTEndothelial-to-hematopoietic transition (EHT) is crucial for hematopoietic stem cell (HSC) generation. During EHT, the morphology of hemogenic endothelial cells (HECs) changes from flat and adherent to spherical HSCs, which detach from the dorsal aorta. HECs attain a rounded shape in a mitosis-independent manner before cell adhesion termination, suggesting an atypical cell-rounding mechanism. However, the direct mechanisms underlying this change in cell morphology during EHT remain unclear. Here, we show that large vacuoles were transiently formed in HECs and that aquaporin-1 (AQP1) was localized in the vacuole and plasma membranes. Overexpression of AQP1 in non-HECs induced ectopic vacuole expansion, cell rounding, and subsequent cell detachment from the endothelium into the bloodstream, mimicking EHT. Loss of redundant AQP functions by CRISPR/Cas9 gene editing in HECs impeded the morphological EHT. Our findings provide the first evidence indicating that morphological segregation of HSCs from endothelial cells is regulated by water influx into vacuoles. These findings provide important insights for further exploration of the mechanisms underlying cell/tissue morphogenesis through water-adoptive cellular responses.SUMMARY STATEMENTHemogenic endothelial cells transiently form large vacuoles during endothelial-to-hematopoietic transition. Aquaporin water channels regulate cell rounding and detachment of emerging hematopoietic stem cells through vacuole formation.
Development of the alpha rhythm is linked to visual white matter pathways and visual detection performance
Authors: Caffarra Sendy , Kanopka Klint , Kruper John , Richie-Halford Adam , Roy Ethan , Rokem Ariel , Yeatman Jason D. 
Published: September 2022

AbstractAlpha is the strongest electrophysiological rhythm in awake humans at rest. Despite its predominance in the EEG signal, strong variations can be observed in alpha properties during development, with an increase of alpha frequency over childhood and adulthood. Here we tested the hypothesis that these changes of alpha rhythm are related to the maturation of visual white matter pathways. We capitalized on a large dMRI-EEG dataset (dMRI n=2,747, EEG n=2,561) of children and adolescents (age range: 5-21 years old) and showed that maturation of the optic radiation specifically accounts for developmental changes of alpha frequency. Behavioral analyses also confirmed that variations of alpha frequency are related to maturational changes in visual perception. The present findings demonstrate the close link between developmental variations in white matter tissue properties, electrophysiological responses, and behavior.