Enhanced activity of hyperthermostable Pyrococcus horikoshii endoglucanase in superbase ionic liquids

Objectives Ionic liquids (ILs) that dissolve biomass are harmful to the enzymes that degrade lignocellulose. Enzyme hyperthermostability promotes a tolerance to ILs. Therefore, the limits of hyperthemophilic Pyrococcus horikoschii endoglucanase (PhEG) to tolerate 11 superbase ILs were explored. Results PhEG was found to be most tolerant to 1-ethyl-3-methylimidazolium acetate ([EMIM]OAc) in soluble 1% carboxymethylcellulose (CMC) and insoluble 1% Avicel substrates. At 35% concentration, this IL caused an increase in enzyme activity (up to 1.5-fold) with CMC. Several ILs were more enzyme inhibiting with insoluble Avicel than with soluble CMC. K-m increased greatly in the presence ILs, indicating significant competitive inhibition. Increased hydrophobicity of the IL cation or anion was associated with the strongest enzyme inhibition and activation. Surprisingly, PhEG activity was increased 2.0-2.5-fold by several ILs in 4% substrate. Cations exerted the main role in competitive inhibition of the enzyme as revealed by their greater binding energy to the active site. Conclusions These results reveal new ways to design a beneficial combination of ILs and enzymes for the hydrolysis of lignocellulose, and the strong potential of PhEG in industrial, high substrate concentrations in aqueous IL solutions.Peer reviewe

Inhibition of hyperthermostable xylanases by superbase ionic liquids

The use of enzymes in aqueous solutions of ionic liquids (ILs) could be useful for the enzymatic treatment of lignocellulose. Hydrophilic ILs that dissolve lignocellulose are harmful to enzymes. The toleration limits and enzyme-friendly superbase IL combinations were investigated for the hyperthermophilic Thermopolyspora flexuosa GH10 xylanase (endo-1,4-beta-xylanase EC 3.2.1.8) TfXYN10A and Dictyoglomus thermophilum GH11 xylanase DtXYN11B. TfXYN10A was more tolerant than DtXYN11B to acetate or propionate-based ILs. However, when the anion of the ILs was bigger (guaiacolate), GH11 xylanase showed higher tolerance to ILs. 1-Ethyl-3-methylimidazolium acetate ([EMIM]OAc), followed by 1,1,3,3-tetramethylguanidine acetate ([TMGH]OAc), were the most enzyme-friendly ILs for TfXYN10A and [TMGH](+)-based ILs were tolerated best by DtXYN11B. Double-ring cations and a large size anion were associated with the strongest enzyme inhibition. Competitive inhibition appears to be a general factor in the reduction of enzyme activity. However, with guaiacolate ILs, the denaturation of proteins may also contribute to the reduction in enzyme activity. Molecular docking with IL cations and anions indicated that the binding mode and shape of the active site affect competitive inhibition, and the cobinding of cations and anions to separate active site positions caused the strongest enzyme inhibition.Peer reviewe

Työterveyshuollon palveluiden käyttö työikäisten terveydenhuollon palvelukokonaisuudessa : Perusterveydenhuollon hoitoilmoitusrekisterin (Avohilmo) tietojen hyödyntäminen tilannekuvan luomisessa

Huolimatta sähköisten potilaskertomuksien laajamittaisesta käytöstä Suomessa, tietopohja työterveyshuollon palveluiden tarjonnasta ja käytöstä on yhä rajallista. Työterveyshuollon käyttötietoja on kerätty valtakunnalliseen Perusterveydenhuollon avohoidon hoitoilmoitusrekisteriin (Avohilmoon) pääsääntöisesti vasta vuoden 2019 alusta lähtien. Työterveyshuollon palveluntuottajat ovat tulleet enenevästi mukaan Avohilmo-tiedonkeruuseen vuosien 2020 ja 2021 aikana, minkä myötä Avohilmo-tietojen kattavuus on hiljalleen parantunut. Vuodesta 2019 alkaen Avohilmoon on kerätty uusia, työ-terveyshuollon toimintaa ja tavoitteita paremmin kuvaavia tietoja. Kerätyt työterveyshuollon käyttötiedot voivat tarjota oleellista lisätietoa työterveyshuollon roolista osana työikäisten terveydenhuollon kokonaisuutta. On kuitenkin yhä epäselvää, mikä on tämän tiedon laatu ja soveltuvuus tutkimuskäyttöön. Tässä tutkimuksessa tarkastelimme työikäisen väestön (15-74-vuotiaat) työterveyshuoltopalveluiden käytöstä Avohilmo-rekisteriin kertyneen yksilötasoisen tiedon laatua vuosina 2020-2022. Arvioimme laatua erityisesti tiedon kattavuuden ja virheettömyyden osalta. Huomioimme tarkaste-lussa laajasti eri palvelu- ja käyntimuotoja. Keskityimme kuvaamaan työterveyshuollon asiakasmääriä, palveluiden käyttöä, henkilöstörakennetta ja lähetemääriä, sekä alueittain että palveluntuottajittain. Lisäksi tarkastelimme millä tavoin työterveyshuollon käyttö jakautuu työterveyshuollon eh-käiseviin palveluihin ja työterveyshuollon sairaanhoitoon ja muuhun terveydenhoitoon, sekä miten työterveyshuollon sairaanhoitoa käytetään julkisen perusterveydenhuollon rinnalla. Olimme erityisen kiinnostuneita työterveyshuollon palveluiden yhteyksistä ja integraatiosta julkiseen terveydenhuoltojärjestelmään. Työpaperi on suunnattu erityisesti alan tutkijoille, päätöksentekijöille, palveluntuottajille sekä Hilmo-tietojen tuotannosta vastaaville henkilöille

Screening of glycoside hydrolases and ionic liquids for fibre modification

BACKGROUNDThis study elaborates the possibility to apply combined ionic liquid (IL) and enzyme treatments for pulp fibre modification. The approach involves swelling of fibre surfaces with IL followed by enzymatic modification of the disrupted fibre surface using carbohydrate active enzymes. RESULTSThe capacity of seven cellulose-dissolving or cellulose-swelling ionic liquids to swell pulp fibres was compared. In addition, thirteen cellulases and five xylanases were screened for their IL tolerance, which determines their applicability in combined or sequential IL-enzyme treatments of fibres. Among the studied ionic liquids, 1-ethyl-3-methylimidazolium dimethylphosphate ([EMIM]DMP) and 1,3-dimethylimidazolium dimethylphosphate ([DMIM]DMP) had the strongest effect on fibre swelling. These solvents were also found to be the least inactivating for the studied enzymes. CONCLUSIONEnzyme compatibility and cellulose-dissolving capability are not two conflicting properties of an ionic liquid. (c) 2017 Society of Chemical IndustryPeer reviewe

Nurses’ ability to timely activate rapid response systems for deteriorating patients: A comparative case scenario study between Finnish and British nurses

Supplementary data to this article can be found online at https://doi.org/10.1016/j.iccn.2020.102871

Oncogenic Herpesvirus Utilizes Stress-Induced Cell Cycle Checkpoints for Efficient Lytic Replication

Kaposi's sarcoma herpesvirus (KSHV) causes Kaposi's sarcoma and certain lymphoproliferative malignancies. Latent infection is established in the majority of tumor cells, whereas lytic replication is reactivated in a small fraction of cells, which is important for both virus spread and disease progression. A siRNA screen for novel regulators of KSHV reactivation identified the E3 ubiquitin ligase MDM2 as a negative regulator of viral reactivation. Depletion of MDM2, a repressor of p53, favored efficient activation of the viral lytic transcription program and viral reactivation. During lytic replication cells activated a p53 response, accumulated DNA damage and arrested at G2-phase. Depletion of p21, a p53 target gene, restored cell cycle progression and thereby impaired the virus reactivation cascade delaying the onset of virus replication induced cytopathic effect. Herpesviruses are known to reactivate in response to different kinds of stress, and our study now highlights the molecular events in the stressed host cell that KSHV has evolved to utilize to ensure efficient viral lytic replication.Peer reviewe

The Glanville fritillary genome retains ancient karyotype and reveals selective chromosomal fusions in Lepidoptera

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Antiviral Properties of Chemical Inhibitors of Cellular Anti-Apoptotic Bcl-2 Proteins

Viral diseases remain serious threats to public health because of the shortage of effective means of control. To combat the surge of viral diseases, new treatments are urgently needed. Here we show that small-molecules, which inhibit cellular anti-apoptotic Bcl-2 proteins (Bcl-2i), induced the premature death of cells infected with different RNA or DNA viruses, whereas, at the same concentrations, no toxicity was observed in mock-infected cells. Moreover, these compounds limited viral replication and spread. Surprisingly, Bcl-2i also induced the premature apoptosis of cells transfected with viral RNA or plasmid DNA but not of mock-transfected cells. These results suggest that Bcl-2i sensitizes cells containing foreign RNA or DNA to apoptosis. A comparison of the toxicity, antiviral activity, and side effects of six Bcl-2i allowed us to select A-1155463 as an antiviral lead candidate. Thus, our results pave the way for the further development of Bcl-2i for the prevention and treatment of viral diseases.Peer reviewe