Keruloplasmiini aikuisten maksasairauksien diagnostiikassa

Tiivistelmä. Wilsonin tauti on harvinainen periytyvä kupariaineenvaihduntahäiriö, joka johtaa kuparin kertymiseen eri kudoksiin keruloplasmiini nimisen proteiinin puuttuessa, joka normaalisti sitoo suurimman osan plasman kuparista. Kuparin kertyminen elimistöön aiheuttaa maksaperäisiä, neurologisia ja psykiatrisia oireita. Wilsonin tautia epäiltäessä ensisijainen tutkimus on seerumin keruloplasmiinin määritys, vaikka se ei olekaan tälle yksinään diagnostinen. Tutkimuksemme tarkoituksena on selvittää Pohjois-Pohjanmaan sairaanhoitopiirin (PPSHP) alueella vuosina 2015–2017 keruloplasmiinimäärityksistä aiheutuneet kustannukset, näytteistä saadut tulokset sekä muita keruloplasmiiniin assosioituvia taustatekijöitä. Aineisto koostui potilaista, joilta on otettu keruloplasmiinimääritys vuosien 2015–2017 välisenä aikana. Aineistosta poistettiin alle 16-vuotiaat sekä ne potilaat, joilla ei ollut potilasasiakirjamerkintöjä siltä hoitojaksolta, jolla keruloplasmiinimääritys tehtiin. Potilaiden tiedot on kerätty Esko-potilastietojärjestelmästä. Kerätty data käsiteltiin siten, että keruloplasmiinin osalta potilaat jaoteltiin viitearvon alapuolella, yläpuolella ja viitealueella oleviin arvoihin. Potilaat jaoteltiin iän perusteella kolmeen ryhmään. Muita potilasasiakirjoista kerättyjä tietoja olivat aiemmat maksasairaudet, keruloplasmiinimäärityksen indikaatio ja löydökset, pyynnön tehnyt yksikkö sekä mahdollinen hoitojakson aikana ohjelmoitu maksabiopsia. Tutkimuksessa selvisi, että keruloplasmiinia otettiin eniten yli 55-vuotiaiden ryhmässä, jossa odotusarvo Wilsonin taudin löytymiselle on alhaisin. Aineistosta ei löytynyt uusia Wilsonin tautia sairastavia. Tämän tutkimuksen kohteena olevan tutkimusjoukon osalta keruloplasmiini-tutkimusta tilattiin eniten erikoissairaanhoidon yksiköissä, joiden osalta lukumääräisesti suurin yksittäinen tilaaja oli Oulun yliopistollisen sairaalan vatsakeskus. Yleisin syy keruloplasmiini-tutkimukselle oli maksasairauksien erotusdiagnostiikka. Suurin osa potilaista, joilta keruloplasmiini oli otettu, jäi ilman maksalöydöksiä. Tutkimuksemme mukaan neurologiset oireet saattavat ennustaa paremmin alhaista keruloplasmiinia kuin maksaperäiset oireet, mutta suppean aineiston vuoksi löydös jää suuntaa antavaksi. Esitämme, että keruloplasmiinin määrittäminen tulisi keskittää erikoissairaanhoidon yksiköihin, joissa poikkeavan keruloplasmiiniarvon merkitsevyys kokonaistilanteen kannalta pystytään paremmin arvioimaan yhteistyössä eri erikoisalojen kanssa ja tarvittaessa järjestämään jatkotutkimuksia kuten esimerkiksi AASLD:n (American Association for the Study of Liver Diseases) suosituksessa

Glycogen Synthase Kinase (GSK) 3β phosphorylates and protects nuclear myosin 1c from proteasome-mediated degradation to activate rDNA transcription in early G1 cells

Nuclear myosin 1c (NM1) mediates RNA polymerase I (pol I) transcription activation and cell cycle progression by facilitating PCAF-mediated H3K9 acetylation, but the molecular mechanism by which NM1 is regulated remains unclear. Here, we report that at early G1 the glycogen synthase kinase (GSK) 3β phosphorylates and stabilizes NM1, allowing for NM1 association with the chromatin. Genomic analysis by ChIP-Seq showed that this mechanism occurs on the rDNA as active GSK3β selectively occupies the gene. ChIP assays and transmission electron microscopy in GSK3β-/- mouse embryonic fibroblasts indicated that at G1 rRNA synthesis is suppressed due to decreased H3K9 acetylation leading to a chromatin state incompatible with transcription. We found that GSK3β directly phosphorylates the endogenous NM1 on a single serine residue (Ser-1020) located within the NM1 C-terminus. In G1 this phosphorylation event stabilizes NM1 and prevents NM1 polyubiquitination by the E3 ligase UBR5 and proteasome-mediated degradation. We conclude that GSK3β-mediated phosphorylation of NM1 is required for pol I transcription activation

Soluble AXL is a novel blood marker for early detection of pancreatic ductal adenocarcinoma and differential diagnosis from chronic pancreatitis

Background: Early diagnosis is crucial for patients with pancreatic ductal adenocarcinoma (PDAC). The AXL receptor tyrosine kinase is proteolytically processed releasing a soluble form (sAXL) into the blood stream. Here we explore the use of sAXL as a biomarker for PDAC. Methods: AXL was analysed by immunohistochemistry in human pancreatic tissue samples. RNA expression analysis was performed using TCGA/GTEx databases. The plasma concentrations of sAXL, its ligand GAS6, and CA19-9 were studied in two independent cohorts, the HMar cohort (n = 59) and the HClinic cohort (n = 142), including healthy controls, chronic pancreatitis (CP) or PDAC patients, and in a familial PDAC cohort (n = 68). AXL expression and sAXL release were studied in PDAC cell lines and murine models. Findings: AXL is increased in PDAC and precursor lesions as compared to CP or controls. sAXL determined in plasma from two independent cohorts was significantly increased in the PDAC group as compared to healthy controls or CP patients. Patients with high levels of AXL have a lower overall survival. ROC analysis of the plasma levels of sAXL, GAS6, or CA19-9 in our cohorts revealed that sAXL outperformed CA19-9 for discriminating between CP and PDAC. Using both sAXL and CA19-9 increased the diagnostic value. These results were validated in murine models, showing increased sAXL specifically in animals developing PDAC but not those with precursor lesions or acinar tumours. Interpretation: sAXL appears as a biomarker for early detection of PDAC and PDAC–CP discrimination that could accelerate treatment and improve its dismal prognosis. Funding: This work was supported by grants PI20/00625 (PN), RTI2018-095672-B-I00 (AM and PGF), PI20/01696 (MG) and PI18/01034 (AC) from MICINN-FEDER and grant 2017/SGR/225 (PN) from Generalitat de Catalunya. © 2021 The Author(s

Interaction of eukaryotic translation initiation factor 4G with the nuclear cap-binding complex provides a link between nuclear and cytoplasmic functions of the m7 guanosine cap

In eukaryotes the majority of mRNAs have an m7G cap that is added cotranscriptionally and that plays an important role in many aspects of mRNA metabolism. The nuclear cap-binding complex (CBC; consisting of CBP20 and CBP80) mediates the stimulatory functions of the cap in pre-mRNA splicing, 3' end formation, and U snRNA export. As little is known about how nuclear CBC mediates the effects of the cap in higher eukaryotes, we have characterized proteins that interact with CBC in HeLa cell nuclear extracts as potential mediators of its function. Using cross-linking and coimmunoprecipitation, we show that eukaryotic translation initiation factor 4G (eIF4G), in addition to its function in the cytoplasm, is a nuclear CBC-interacting protein. We demonstrate that eIF4G interacts with CBC in vitro and that, in addition to its cytoplasmic localization, there is a significant nuclear pool of eIF4G in mammalian cells in vivo. Immunoprecipitation experiments suggest that, in contrast to the cytoplasmic pool, much of the nuclear eIF4G is not associated with eIF4E (translation cap binding protein of eIF4F) but is associated with CBC. While eIF4G stably associates with spliceosomes in vitro and shows close association with spliceosomal snRNPs and splicing factors in vivo, depletion studies show that it does not participate directly in the splicing reaction. Taken together the data indicate that nuclear eIF4G may be recruited to pre-mRNAs via its interaction with CBC and accompanies the mRNA to the cytoplasm, facilitating the switching of CBC for eIF4F. This may provide a mechanism to couple nuclear and cytoplasmic functions of the mRNA cap structure

London 2012: changing delivery patterns in response to the impact of the Games on traffic flows

The paper addresses road freight transport operations during the London Olympic and Paralympic Games in 2012. It presents work carried out prior to the Games to understand pre-Games patterns of freight deliveries in London (for both light and heavy goods vehicles) and the results of modelling work carried out to assess the likely impacts of the Games road restrictions on freight operations. The modelling results indicated that increases in total hours travelled carrying out collection and delivery work would range from 1.4% to 11.4% in the six sectors considered. The results suggested increases in hours travelled in excess of 3.5% in four of the six sectors modelled. The possible actions that could be taken by organizations to reduce these negative impacts were also modelled and the results indicated that such actions would help to mitigate the impact of the road restrictions imposed on operators during the Games. The actual impacts of the 2012 Games on transport both in general terms and specifically in terms of freight transport are also discussed, together with the success of the actions taken by Transport for London (TfL) to help the road freight industry. The potential freight transport legacy of the London 2012 Games in terms of achieving more sustainable urban freight transport is considered and the steps being taken by TfL to help ensure that such a legacy can be realized are discussed. Such steps include policy-makers continuing to collaborate closely with the freight industry through the ‘London Freight Forum’, and TfL's efforts to encourage and support companies revising their delivery and collection times to the off-peak; improving freight planning in the design and management of TfL-funded road schemes; electronic provision of traffic information by TfL to the freight industry, and the further development of freight journey planning tools

An HDAC9-MALAT1-BRG1 complex mediates smooth muscle dysfunction in thoracic aortic aneurysm

Thoracic aortic aneurysm (TAA) has been associated with mutations affecting members of the TGF-β signaling pathway, or components and regulators of the vascular smooth muscle cell (VSMC) actomyosin cytoskeleton. Although both clinical groups present similar phenotypes, the existence of potential common mechanisms of pathogenesis remain obscure. Here we show that mutations affecting TGF-β signaling and VSMC cytoskeleton both lead to the formation of a ternary complex comprising the histone deacetylase HDAC9, the chromatin-remodeling enzyme BRG1, and the long noncoding RNA MALAT1. The HDAC9–MALAT1–BRG1 complex binds chromatin and represses contractile protein gene expression in association with gain of histone H3-lysine 27 trimethylation modifications. Disruption of Malat1 or Hdac9 restores contractile protein expression, improves aortic mural architecture, and inhibits experimental aneurysm growth. Thus, we highlight a shared epigenetic pathway responsible for VSMC dysfunction in both forms of TAA, with potential therapeutic implication for other known HDAC9-associated vascular diseases

A kinetic and thermodynamic investigation into the removal of methyl orange from wastewater utilizing fly ash in different process configurations

The removal of methyl orange using coal fly ash, which is a widely available low-cost adsorbent, has been investigated. Adsorption studies for dye removal were conducted using various configurations such as batch, column and heap adsorption at various temperatures and adsorbent dosages at neutral pH. The Langmuir, Freundlich and Tempkin isotherm models were used to describe the process. The Freundlich model best represented the adsorption. Kinetic studies show the adsorption followed pseudo-second-order kinetics. Thermodynamic studies show that the process is spontaneous, endothermic and random. Column configuration was found to be the most efficient with a dye removal percentage of 99.95%, followed by heap adsorption at 99.25% removal and lastly batch configuration with 96.68% removal. Economic analysis shows that column operation would be the most effective for practical implementation

Towards the prediction of essential genes by integration of network topology, cellular localization and biological process information

<p>Abstract</p> <p>Background</p> <p>The identification of essential genes is important for the understanding of the minimal requirements for cellular life and for practical purposes, such as drug design. However, the experimental techniques for essential genes discovery are labor-intensive and time-consuming. Considering these experimental constraints, a computational approach capable of accurately predicting essential genes would be of great value. We therefore present here a machine learning-based computational approach relying on network topological features, cellular localization and biological process information for prediction of essential genes.</p> <p>Results</p> <p>We constructed a decision tree-based meta-classifier and trained it on datasets with individual and grouped attributes-network topological features, cellular compartments and biological processes-to generate various predictors of essential genes. We showed that the predictors with better performances are those generated by datasets with integrated attributes. Using the predictor with all attributes, i.e., network topological features, cellular compartments and biological processes, we obtained the best predictor of essential genes that was then used to classify yeast genes with unknown essentiality status. Finally, we generated decision trees by training the J48 algorithm on datasets with all network topological features, cellular localization and biological process information to discover cellular rules for essentiality. We found that the number of protein physical interactions, the nuclear localization of proteins and the number of regulating transcription factors are the most important factors determining gene essentiality.</p> <p>Conclusion</p> <p>We were able to demonstrate that network topological features, cellular localization and biological process information are reliable predictors of essential genes. Moreover, by constructing decision trees based on these data, we could discover cellular rules governing essentiality.</p

Genome Analysis Reveals Interplay between 5′UTR Introns and Nuclear mRNA Export for Secretory and Mitochondrial Genes

In higher eukaryotes, messenger RNAs (mRNAs) are exported from the nucleus to the cytoplasm via factors deposited near the 5′ end of the transcript during splicing. The signal sequence coding region (SSCR) can support an alternative mRNA export (ALREX) pathway that does not require splicing. However, most SSCR–containing genes also have introns, so the interplay between these export mechanisms remains unclear. Here we support a model in which the furthest upstream element in a given transcript, be it an intron or an ALREX–promoting SSCR, dictates the mRNA export pathway used. We also experimentally demonstrate that nuclear-encoded mitochondrial genes can use the ALREX pathway. Thus, ALREX can also be supported by nucleotide signals within mitochondrial-targeting sequence coding regions (MSCRs). Finally, we identified and experimentally verified novel motifs associated with the ALREX pathway that are shared by both SSCRs and MSCRs. Our results show strong correlation between 5′ untranslated region (5′UTR) intron presence/absence and sequence features at the beginning of the coding region. They also suggest that genes encoding secretory and mitochondrial proteins share a common regulatory mechanism at the level of mRNA export