Hepatocyte nuclear factor-1β (HNF-1β) is a tissue-specific transcription factor that is required for normal kidney development and renal epithelial differentiation. Mutations of HNF-1β produce congenital kidney abnormalities and inherited renal tubulopathies. Here, we show that ablation of HNF-1β in mIMCD3 renal epithelial cells results in activation of β-catenin and increased expression of lymphoid enhancer–binding factor 1 (LEF1), a downstream effector in the canonical Wnt signaling pathway. Increased expression and nuclear localization of LEF1 are also observed in cystic kidneys from Hnf1b mutant mice. Expression of dominant-negative mutant HNF-1β in mIMCD3 cells produces hyperresponsiveness to exogenous Wnt ligands, which is inhibited by siRNA-mediated knockdown of Lef1. WT HNF-1β binds to two evolutionarily conserved sites located 94 and 30 kb from the mouse Lef1 promoter. Ablation of HNF-1β decreases H3K27 trimethylation repressive marks and increases β-catenin occupancy at a site 4 kb upstream to Lef1. Mechanistically, WT HNF-1β recruits the polycomb-repressive complex 2 that catalyzes H3K27 trimethylation. Deletion of the β-catenin–binding domain of LEF1 in HNF-1β–deficient cells abolishes the increase in Lef1 transcription and decreases the expression of downstream Wnt target genes. The canonical Wnt target gene, Axin2, is also a direct transcriptional target of HNF-1β through binding to negative regulatory elements in the gene promoter. These findings demonstrate that HNF-1β regulates canonical Wnt target genes through long-range effects on histone methylation at Wnt enhancers and reveal a new mode of active transcriptional repression by HNF-1β.

Thoracic great vessels such as the aorta and subclavian arteries are formed through dynamic remodeling of embryonic pharyngeal arch arteries (PAAs). Previous work has shown that loss of a basic helix-loop-helix transcription factor Hey1 in mice causes abnormal fourth PAA development and lethal great vessel anomalies resembling congenital malformations in humans. However, how Hey1 mediates vascular formation remains unclear. In this study, we revealed that Hey1 in vascular endothelial cells, but not in smooth muscle cells, played essential roles for PAA development and great vessel morphogenesis in mouse embryos. Tek-Cre–mediated Hey1 deletion in endothelial cells affected endothelial tube formation and smooth muscle differentiation in embryonic fourth PAAs and resulted in interruption of the aortic arch and other great vessel malformations. Cell specificity and signal responsiveness of Hey1 expression were controlled through multiple cis-regulatory regions. We found two distal genomic regions that had enhancer activity in endothelial cells and in the pharyngeal epithelium and somites, respectively. The novel endothelial enhancer was conserved across species and was specific to large-caliber arteries. Its transcriptional activity was regulated by Notch signaling in vitro and in vivo, but not by ALK1 signaling and other transcription factors implicated in endothelial cell specificity. The distal endothelial enhancer was not essential for basal Hey1 expression in mouse embryos but may likely serve for Notch-dependent transcriptional control in endothelial cells together with the proximal regulatory region. These findings help in understanding the significance and regulation of endothelial Hey1 as a mediator of multiple signaling pathways in embryonic vascular formation.

Investigations of bacterial resistance strategies can aid in the development of new antimicrobial drugs as a countermeasure to the increasing worldwide prevalence of bacterial antibiotic resistance. One such strategy involves the TipA class of transcription factors, which constitute minimal autoregulated multidrug resistance (MDR) systems against diverse antibiotics. However, we have insufficient information regarding how antibiotic binding induces transcriptional activation to design molecules that could interfere with this process. To learn more, we determined the crystal structure of SkgA from Caulobacter crescentus as a representative TipA protein. We identified an unexpected spatial orientation and location of the antibiotic-binding TipAS effector domain in the apo state. We observed that the α6–α7 region of the TipAS domain, which is canonically responsible for forming the lid of antibiotic-binding cleft to tightly enclose the bound antibiotic, is involved in the dimeric interface and stabilized via interaction with the DNA-binding domain in the apo state. Further structural and biochemical analyses demonstrated that the unliganded TipAS domain sterically hinders promoter DNA binding but undergoes a remarkable conformational shift upon antibiotic binding to release this autoinhibition via a switch of its α6–α7 region. Hence, the promoters for MDR genes including tipA and RNA polymerases become available for transcription, enabling efficient antibiotic resistance. These insights into the molecular mechanism of activation of TipA proteins advance our understanding of TipA proteins, as well as bacterial MDR systems, and may provide important clues to block bacterial resistance.

Cellular energy demands are met by uptake and metabolism of nutrients like glucose. The principal transcriptional regulator for adapting glycolytic flux and downstream pathways like de novo lipogenesis to glucose availability in many cell types is carbohydrate response element–binding protein (ChREBP). ChREBP is activated by glucose metabolites and post-translational modifications, inducing nuclear accumulation and regulation of target genes. Here we report that ChREBP is modified by proline hydroxylation at several residues. Proline hydroxylation targets both ectopically expressed ChREBP in cells and endogenous ChREBP in mouse liver. Functionally, we found that specific hydroxylated prolines were dispensable for protein stability but required for the adequate activation of ChREBP upon exposure to high glucose. Accordingly, ChREBP target gene expression was rescued by re-expressing WT but not ChREBP that lacks hydroxylated prolines in ChREBP-deleted hepatocytes. Thus, proline hydroxylation of ChREBP is a novel post-translational modification that may allow for therapeutic interference in metabolic diseases.

The Wnt/β-catenin pathway is one of the major pathways that regulates embryonic development, adult homeostasis, and stem cell self-renewal. In this pathway, transcription factors T-cell factor and lymphoid enhancer factor (TCF/LEF) serve as a key switch to repress or activate Wnt target gene transcription by recruiting repressor molecules or interacting with the β-catenin effector, respectively. It has become evident that the protein stability of the TCF/LEF family members may play a critical role in controlling the activity of the Wnt/β-catenin signaling pathway. However, factors that regulate the stability of TCF/LEFs remain largely unknown. Here, we report that pVHL binding protein 1 (VBP1) regulates the Wnt/β-catenin signaling pathway by controlling the stability of TCF/LEFs. Surprisingly, we found that either overexpression or knockdown of VBP1 decreased Wnt/β-catenin signaling activity in both cultured cells and zebrafish embryos. Mechanistically, VBP1 directly binds to all four TCF/LEF family members and von Hippel-Lindau tumor-suppressor protein (pVHL). Either overexpression or knockdown of VBP1 increases the association between TCF/LEFs and pVHL and then decreases the protein levels of TCF/LEFs via proteasomal degradation. Together, our results provide mechanistic insights into the roles of VBP1 in controlling TCF/LEFs protein stability and regulating Wnt/β-catenin signaling pathway activity.

Mitochondrial dysfunction is associated with a variety of human diseases including neurodegeneration, diabetes, nonalcohol fatty liver disease (NAFLD), and cancer, but its underlying causes are incompletely understood. Using the human hepatic cell line HepG2 as a model, we show here that endoplasmic reticulum-associated degradation (ERAD), an ER protein quality control process, is critically required for mitochondrial function in mammalian cells. Pharmacological inhibition or genetic ablation of key proteins involved in ERAD increased cell death under both basal conditions and in response to proinflammatory cytokines, a situation frequently found in NAFLD. Decreased viability of ERAD-deficient HepG2 cells was traced to impaired mitochondrial functions including reduced ATP production, enhanced reactive oxygen species (ROS) accumulation, and increased mitochondrial outer membrane permeability. Transcriptome profiling revealed widespread down-regulation of genes underpinning mitochondrial functions, and up-regulation of genes associated with tumor growth and aggression. These results highlight a critical role for ERAD in maintaining mitochondrial functional and structural integrity and raise the possibility of improving cellular and organismal mitochondrial function via enhancing cellular ERAD capacity.

Acute lung injury (ALI), is a rapidly progressing heterogenous pulmonary disorder that possesses a high risk of mortality. Accumulating evidence has implicated the activation of the p65 subunit of NF-κB [NF-κB(p65)] activation in the pathological process of ALI. microRNAs (miRNAs), a group of small RNA molecules, have emerged as major governors due to their post-transcriptional regulation of gene expression in a wide array of pathological processes, including ALI. The dysregulation of miRNAs and NF-κB activation has been implicated in human diseases. In the current study, we set out to decipher the convergence of miR-99b and p65 NF-κB activation in ALI pathology. We measured the release of pro-inflammatory cytokines (IL-1β, IL-6, and TNFα) in bronchoalveolar lavage fluid using ELISA. MH-S cells were cultured and their viability were detected with cell counting kit 8 (CCK8) assays. The results showed that miR-99b was up-regulated, while PRDM1 was down-regulated in a lipopolysaccharide (LPS)-induced murine model of ALI. Mechanistic investigations showed that NF-κB(p65) was enriched at the miR-99b promoter region, and further promoted its transcriptional activity. Furthermore, miR-99b targeted PRDM1 by binding to its 3'UTR, causing its down-regulation. This in-creased lung injury, as evidenced by increased wet/dry ratio of mouse lung, myeloperoxidase activity and pro-inflammatory cytokine secretion, and enhanced infiltration of inflammatory cells in lung tissues. Together, our findings indicate that NF-κB(p65) promotion of miR-99b can aggravate ALI in mice by down-regulating the expression of PRDM1.

Paramphistomosis, caused by the rumen fluke, Calicophoron daubneyi, is a parasitic infection of ruminant livestock which has seen a rapid rise in prevalence throughout Western Europe in recent years. Following ingestion of metacercariae (parasite cysts) by the mammalian host, newly-excysted juveniles (NEJs) emerge and invade the duodenal submucosa which causes significant pathology in heavy infections. The immature larvae then migrate upwards, along the gastrointestinal tract, and enter the rumen where they mature and begin to produce eggs. Despite their emergence, and sporadic outbreaks of acute disease, we know little about the molecular mechanisms used by C. daubneyi to establish infection, acquire nutrients and to avoid the host immune response. Here, transcriptome analysis of four intra-mammalian life-cycle stages, integrated with secretome analysis of the NEJ and adult parasites (responsible for acute and chronic disease respectively), revealed how the expression and secretion of selected families of virulence factors and immunomodulators are regulated in accordance with fluke development and migration. Our data show that whilst a family of cathepsins B with varying S2 sub-site residues (indicating distinct substrate specificities) are differentially secreted by NEJs and adult flukes, cathepsins L and F are secreted in low abundance by NEJs only. We found that C. daubneyi has an expanded family of aspartic peptidases, which is up-regulated in adult worms, although they are underrepresented in the secretome. The most abundant proteins in adult fluke secretions were helminth defence molecules (HDMs) that likely establish an immune environment permissive to fluke survival and/or neutralise pathogen-associated molecular patterns (PAMPs) such as bacterial lipopolysaccharide in the microbiome-rich rumen. The distinct collection of molecules secreted by C. daubneyi allowed the development of the first coproantigen-based ELISA for paramphistomosis which, importantly, did not recognise antigens from other helminths commonly found as co-infections with rumen fluke.

In nucleotide excision repair, bulky DNA lesions such as UV-induced cyclobutane pyrimidine dimers are removed from the genome by concerted dual incisions bracketing the lesion, followed by gap filling and ligation. So far, two dual-incision patterns have been discovered: the prokaryotic type, which removes the damage in 11–13-nucleotide-long oligomers, and the eukaryotic type, which removes the damage in 24–32-nucleotide-long oligomers. However, a recent study reported that the UvrC protein of Mycobacterium tuberculosis removes damage in a manner analogous to yeast and humans in a 25-mer oligonucleotide arising from incisions at 15 nt from the 3´ end and 9 nt from the 5´ end flanking the damage. To test this model, we used the in vivo excision assay and the excision repair sequencing genome-wide repair mapping method developed in our laboratory to determine the repair pattern and genome-wide repair map of Mycobacterium smegmatis. We find that M. smegmatis, which possesses homologs of the Escherichia coli uvrA, uvrB, and uvrC genes, removes cyclobutane pyrimidine dimers from the genome in a manner identical to the prokaryotic pattern by incising 7 nt 5´ and 3 or 4 nt 3´ to the photoproduct, and performs transcription-coupled repair in a manner similar to E. coli.

VOLUME 289 (2014) PAGES 6604–6618In Fig. 4G, in the foxi1 panel, the images in Fig. 4G, i and l, corresponding to “smad1 MO” and “smad5 MO + samd1/9 mRNA” samples, respectively, were inadvertently reused during figure preparation. This error has now been corrected using images pertaining to each treatment and sample. This correction does not affect the results or conclusions of the work.jbc;295/52/18650/F4F1F4Figure 4G.

GABAergic inhibitory interneurons comprise many subtypes that differ in their molecular, anatomical, and functional properties. In mouse visual cortex, they also differ in their modulation with an animal’s behavioral state, and this state modulation can be predicted from the first principal component (PC) of the gene expression matrix. Here, we ask whether this link between transcriptome and state-dependent processing generalizes across species. To this end, we analysed seven single-cell and single-nucleus RNA sequencing datasets from mouse, human, songbird, and turtle forebrains. Despite homology at the level of cell types, we found clear differences between transcriptomic PCs, with greater dissimilarities between evolutionarily distant species. These dissimilarities arise from two factors: divergence in gene expression within homologous cell types and divergence in cell-type abundance. We also compare the expression of cholinergic receptors, which are thought to causally link transcriptome and state modulation. Several cholinergic receptors predictive of state modulation in mouse interneurons are differentially expressed between species. Circuit modelling and mathematical analyses suggest conditions under which these expression differences could translate into functional differences.

OM EAST responds to the need for Christian books in the Georgian language.

Penn State Health will collect unwanted, unneeded or expired medications, needles and syringes for safe disposal on Saturday, Oct. 26, as part of National Prescription Drug Take-Back Day.

Books played an important role in Aslan's salvation. Now he provides literature to other Central Asians.

Delaware will hold its 22nd National DEA Prescription Drug Take-Back Day on Saturday, April 30, 2022. Delawareans can discard their expired or unused medications at locations statewide between 10:00 a.m. and 2:00 p.m. There will also be Sharps disposals for needle disposal at select locations, and overdose response education with free Narcan available at select locations.

*Editor/Reporter note: We originally stated that five DEA National Prescription Drug Take Back locations would be offering Overdose Response Training and Narcan distribution to the public; there are only two – Milford and Middletown. We also erroneously stated that there are 23 locations currently participating in the National Prescription Drug Take Back event; there are 22.    […]

The Delaware Division of Public Health (DPH) recently held its 24th Prescription Drug Take-Back Day event. Organized nationally by the Drug Enforcement Administration (DEA), Prescription Drug Take-Back Day is operated locally by DPH. The twice-a-year event (April and October) aims to reduce the risk of prescription medications being diverted for misuse and has resulted in 114,470 pounds of medication being collected in Delaware since 2010. […]

The Delaware Division of Public Health (DPH), in collaboration with the Delaware Division of Professional Regulation (DPR), announces important updates to the Mental Health and Substance Use Disorder section of the My Healthy Community (MHC) Dashboard.  These updates are located within this section’s Prescription Monitoring Program (PMP) tab and present trends in prescription drug dispensing in Delaware. The […]

NEW CASTLE – The Delaware Department of Health and Social Services (DHSS) announced today enhancements to the CostAware website, designed to help Delawareans understand how their health care dollars are spent by comparing the variation of average costs for different episodes of care and medical services based on actual medical and pharmacy claims in Delaware. […]

Amidst a shortage of prescription stimulants, concerns have been raised about the potential increase in illicit drug use. With many individuals relying on prescription stimulants to manage conditions such as attention-deficit/hyperactivity disorder (ADHD), the shortage has left many struggling to access the medication they need. This has led some individuals to turn to illicit drugs […]

Delaware will hold its 25th National DEA Prescription Drug Take-Back Day on Saturday, October 28, 2023. Delawareans can discard their expired or unused medications at locations statewide between 10:00 a.m. and 2:00 p.m. Sharps disposals for needle disposal will be available at select locations, as well as overdose response education with free Narcan available at […]

Here Mudassar Khan has explained

Here Mudassar Khan has explained

Here Mudassar Khan has explained

Here Mudassar Khan has explained

Here Mudassar Khan has explained with an example, how to disable Browser Back Button after Logout using JavaScript in ASP.Net MVC.

For Immediate Release: August 13, 2024 Contact: Samuel Barry, Policy Advisor (302) 382-0651, Samuel.Barry@delaware.gov The tracker is a publicly accessible tool for Delawareans to learn about how Prescription Opioid Abatement and Remediation Grants have been distributed to date. Dover, DE – Delaware Auditor of Accounts Lydia E. York published an official Prescription Opioid Settlement Tracker […]

Hello Community,

I have encountered an issue that is a mystery to me and hope somebody could give me a clue about what is happening in Cadence and maybe even a solution?

I am running a test scripted in a SKILL file that sequentially opens two different projects with MC analyses and in between I get an error message box and also multiple logs in CIW with exactly the same text.

Both projects run a simulation with a call like this:

historyName = maeRunSimulation(?session sessionName ?waitUntilDone t)

After this the script closes the current project, opens the next project and executes the same line with maeRunSimulation() for the second project. Then immediately this error message happens, and also is logged repeatedly in the CIW window

The message box looks like this:

The logs I get in CIW:

nil
hiCancelProgressBox(_axlNetlistCreateProgressBar)
nil
hiCancelProgressBox(_axlUILoadForm)
nil
when(dwindow('axlDataViewessWindow1) hiMapWindow(dwindow('axlDataViewessWindow1)))
t
when(dwindow('axlRunSummaryessWindow1) hiMapWindow(dwindow('axlRunSummaryessWindow1)))
t
ERROR (ASSEMBLER-1600): Cannot find an active session named fnxSession0.
You can only modify an ADE Assembler session that is active.
Perhaps the session name was misspelled or has not yet been created.
Verify the session name matches an existing ADE Assembler session.

1>
ERROR (ASSEMBLER-1600): Cannot find an active session named fnxSession0.
You can only modify an ADE Assembler session that is active.
Perhaps the session name was misspelled or has not yet been created.
Verify the session name matches an existing ADE Assembler session.

*WARNING* hiDisplayAppDBox: modal dbox 'adexlMessageDialog' is already displayed!
ERROR (ASSEMBLER-1600): Cannot find an active session named fnxSession0.
You can only modify an ADE Assembler session that is active.
Perhaps the session name was misspelled or has not yet been created.
Verify the session name matches an existing ADE Assembler session.

*WARNING* hiDisplayAppDBox: modal dbox 'adexlMessageDialog' is already displayed!
ERROR (ASSEMBLER-1600): Cannot find an active session named fnxSession0.
You can only modify an ADE Assembler session that is active.
Perhaps the session name was misspelled or has not yet been created.
Verify the session name matches an existing ADE Assembler session.

I would like to send an update about a vmanager regression status x days after the regression has been run. In the current environment, the vmanager regression is creating a new filepath for logs automatically based on regression name/date, so I can't use a cron job to gather logs, as the log location is not known. 

I tried to use the pre session script to launch a detached shell script that would run after a delay, but when the pre_script runs, it waits until everything is completed before finishing and moving on to starting the regression.

Here is the test pre_script I am using:

#!/bin/sh

echo "pre_script start"

delay_script "FIRST" 1
nohup delay_script "SECOND" 30 & disown
delay_script "THIRD" 1

echo "pre_script end"
exit 0

Here is the test delay_script I am using:

#!/bin/sh

echo "Starting $1"

sleep $2

echo "Ending $1"

Here is the script output when run from terminal. After the "pre_script end", I get control back.

Here is the script output when run from vmanager. There is no "nohup", and the pre_session phase doesn't complete until all the delay scripts complete.

My question is, is there a better way to achieve my goal here? (The goal being to run a script from the vmanager log directory automatically x days after the regression). I think I could use the pre_script to send directory information for an auxiliary cron job to pick up, but I would prefer to not have to have extra cronjobs needed for this.

Hello Team,
I am currently following this AWR script posted on them, to run a python script directly from inside AWR using VBA script.

Launching Python from AWRDE VBA Script - AWR Scripts - AWR Knowledgebase

Following all the basic steps with no changes to the code. I have Vscode and python-3.12.2 installed in my system. 

This is the error I am getting while running this code. 

Thank you for your help 

Best Regards

SID

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Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.

Check here each week to keep up with the latest from John MacArthur's pulpit at Grace Community Church.