AAV-mediated and pharmacological induction of Hsp70 expression stimulates survival of retinal ganglion cells following axonal injury.

We evaluated the effect of AAV2- and 17-AAG (17-N-allylamino-17-demethoxygeldanamycin)-mediated upregulation of Hsp70 expression on the survival of retinal ganglion cells (RGCs) injured by optic nerve crush (ONC). AAV2-Hsp70 expression in the retina was primarily observed in the ganglion cell layer. Approximately 75% of all transfected cells were RGCs. RGC survival in AAV2-Hsp70-injected animals was increased by an average of 110% 2 weeks after the axonal injury compared with the control. The increase in cell numbers was not even across the retinas with a maximum effect of approximately 306% observed in the inferior quadrant. 17-AAG-mediated induction of Hsp70 expression has been associated with cell protection in various models of neurodegenerative diseases. We show here that a single intravitreal injection of 17-AAG (0.2 ug ul(-1)) results in an increased survival of ONC-injured RGCs by approximately 49% compared with the vehicle-treated animals. Expression of Hsp70 in retinas of 17-AAG-treated animals was upregulated approximately by twofold compared with control animals. Our data support the idea that the upregulation of Hsp70 has a beneficial effect on the survival of injured RGCs, and the induction of this protein could be viewed as a potential neuroprotective strategy for optic neuropathies

Kestävän kehityksen toteutuminen Suomen lukioissa opiskelijan näkökulmasta

Tiivistelmä. Kestävän kehityksen katsotaan olevan yksi ihmiskunnan suurimpia haasteita ja koulutus on avainasemassa tavoitteiden saavuttamisessa. Koulutuksen tulee vastata yhteiskunnallisiin muutoksiin ja arvoihin. Lukion opetussuunnitelman arvoperustassa peräänkuulutetaan kestävän kehityksen olevan yhteinen ongelma. Sen tulee näkyä toimintakulttuurissa ja kaikkien oppiaineiden opetuksessa. Tässä tutkielmassa selvitetään, miten kestävä kehitys näkyy lukioissa oppitunneilla ja toimintakulttuurissa opiskelijoiden näkökulmasta. Lisäksi tarkastellaan, miten opiskelijat toivovat kestävää kehitystä käsiteltävän. Huomionarvoista on myös se, millainen asenne opiskelijoilla on kestävää kehitystä kohtaan ja miten asenne vaikuttaa muihin vastauksiin. Tutkielman aineisto on kerätty keväällä 2018 yhteistyössä kolmen eri lukion kanssa. Aineisto koostuu 449 opiskelijan kyselyvastauksesta ja kolmesta puolistrukturoidusta ryhmähaastattelusta, joissa jokaisessa oli neljä opiskelijaa. Kyselyaineiston avointen kysymysten vastaukset siirrettiin Microsoft Word -tekstinkäsittelyohjelmaan ja analysoitiin kvantifioimalla. Näin saadut luokat esitetään taulukkomuotoisina. Suljetut kysymykset analysoitiin Microsoft Excel -taulukkolaskentaohjelman avulla ja näin saatiin aikaan havainnollistavia kuvaajia. Litteroitu haastatteluaineisto analysoitiin sisällönanalyysin avulla. Tulokset osoittavat, että kestävä kehitys on edelleen luonnontieteelliseksi mielletty ongelma ja se näkyy eniten biologian ja maantieteen oppitunneilla. Muidenkin oppiaineiden oppitunneilla aihetta käsitellään lähinnä ympäristöllisistä näkökulmista. Toimintakulttuurissa kestävä kehitys ei selkeän enemmistön mielestä näy lainkaan. Opiskelijat toivovat kestävää kehitystä käsiteltävän uusista näkökulmista, mutta perinteisillä opetusmetodeilla. Lisäksi he haluaisivat oppia konkreettisia toimia kestävän kehityksen edistämiseksi. Opiskelijat pitävät kestävää kehitystä tärkeänä ja vastaajien asenteiden havaittiin vaikuttavan vastauksiin. Mitä tärkeämpänä opiskelija pitää kestävää kehitystä, sen paremmin hän osaa määritellä sen, havaitsee sen näkyvän lukioissa ja tuo esiin kehitysehdotuksia. Tutkielma osoittaa, ettei kestävä kehitys näy lukioissa odotetulla tavalla. Tähän ongelmaan tulisi vastata nykyistä tehokkaammalla suunnitelmallisuudella siitä, kenen vastuulla kestävän kehityksen mikäkin osa-alue on. Lisäksi kestävän kehityksen sisällyttäminen kaikkien aineiden opettajien koulutukseen ja täydennyskoulutukseen voisi edesauttaa tilannetta

Pengaruh Kesehatan, Pelatihan Dan Penggunaan Alat Pelindung Diri Terhadap Kecelakaan Kerja Pada Pekerja Konstruksi Di Kota Tomohon

Faktor sumber daya manusia memegang peranan penting dan menentukan sukses atau tidaknya suatu pekerjaan konstruksi. Masalah keselamatan kerja pada pekerja konstruksi masih kurang diperhatikan, masih banyak pekerjaan konstruksi yang mengabaikan faktor keselamatan kerja seperti tidak menggunakan alat pelindung diri. Tujuan dilaksanakannya penelitian ini adalah untuk menganalisa pengaruh kesehatan, pelatihan dan penggunaan alat pelindung diri terhadap kecelakaan kerja pada pekerja konstruksi di kota Tomohon.Metode survey digunakan untuk mendapatkan data utama, dengan cara menyebarkan kuesioner untuk diisi oleh para pekerja konstruksi di kota Tomohon. Populasi penelitian ini sebanyak 3314 pekerja konstruksi, dengan sampel 370 pekerja. Data dianalisa dengan menggunakan uji korelasi dan analisis regresi linier berganda.Hasil penelitian menunjukkan bahwa faktor kesehatan, pelatihan dan penggunaan alat pelindung diri memiliki hubungan negatif dengan kecelakaan kerja, dimana semakin meningkatnya nilai ketiga faktor ini akan menurunkan potensi terjadinya kecelakaan kerja pada pekerja konstruksi di kota Tomohon

Implementation of GDPR: Learning with a local administration case study

The General Data Protection Regulation has come into force in the European Union in May 2018 in order to meet current challenges related to personal data protection and to help harmonise the data protection across the EU. Although the GDPR was expected to benefit companies, being private or public, by offering consistency in data protection activities and liabilities across the EU countries and by enabling more integrated EU wide data protection policies, it poses new challenges to companies. However, if we take a step back and think that this regulation has been in transit for more than 2 years, and that only after the implementation of this regulation has begun the real concern is: are companies ready to make this leap?This work has been supported by national funds through FCT – Fundação para a Ciência e Tecnologia within the Project Scope: UID/CEC/00319/2019

Dramapedagoške tehnike u razvijanju govora u nastavi njemačkog kao stranog jezika

Das Thema dieser Arbeit sind dramapädagogische Techniken zur Förderung des Sprechens im Deutschunterricht als Fremdsprache. Die Arbeit besteht aus zwei Haupteilen, dem theoretischen und dem empirischen Teil. Im theoretischen Teil bespricht man die Grundbedeutung „Dramapädagogik“ und ihre historische Entwicklung, führt die Versuche ihrer Bedeutungsbestimmung und ihre Verbindung mit Theater auf und gibt konkrete Beschreibungen von dramapädagogischen Aufgaben zur „Aufwärmung“ und dramapädagogische Techniken im Unterricht. Der empirische Teil beginnt mit einer Einleitung in die Aktionsforschung und ihren Zweck. Darauffolgend wird die Methodologie der Aktionsforschung vorgestellt, ihre Resultate und die Diskussion über die gewonnenen Daten. Das Hauptziel der Aktionsforschung war die Förderung des Sprechens bei Schülern auf Deutsch durch die Anwendung von dramapädagogischen Techniken. Im Einklang mit den Merkmalen und Charakteristiken der Aktionsforschungen wurde eine Unterrichtstunde entworfen, in der Länge von 45 Minuten, gedacht für die zweite Klasse einer Mittelschule. Die Resultate der Aktionsforschung zeigen, dass die dramapädagogischen Aktivitäten den Schülern interessant sind und ihre Motivation zum aktiven Sprechen im Fremdsprachenunterricht fördern. Die Schüler kamen in Situationen, die nicht nur ihre geistig- sprachlichen Fähigkeiten erfordern, sondern auch die Körpersprache, gemeinsame Aushandlung von Bedeutung und emotionales Verständnis. Durch die Anwendung und Realisierung des dramapädagogischen Zugangs wird das Erlernen einer Fremdsprache auf ein höheres Niveau gebracht, weil es unter anderem die Phantasie, Kreativität, Spontanität und Originalität der Schüler weckt. Somit kann man darauf schließen, dass der dramapädagogische Zugang viel mehr Vorteile aufweist, als der herkömmliche Unterricht, der auf Inhalt und Fakten basiert.Tema ovog rada su dramapedagoške tehnike u procesu razvoja govora u nastavi njemačkog kao stranog jezika. Rad je podijeljen u dvije osnovne cjeline, teorijski i empirijski dio. U teorijskom dijelu rada polazi se od temeljnog pojma „Dramapädagogik“, povijesnog razvoja ove metode, pokušaja definiranja pojma, pojašnjenja temelja dramapedagogije te njezinih poveznica s kazalištem i nastavom stranog jezika. Navedeni dio završava opisom konkretnih dramapedagoških vježbi za „zagrijavanje“ i dramapedagoških tehnika u nastavi. U empirijskom se dijelu daje uvod u akcijsko istraživanje i njegovu svrhu, daje opis metodologije akcijskog istraživanja, prikazuju se rezultati i diskutira o dobivenim podatcima. Osnovni je cilj istraživanja bio unaprijediti i potaknuti razvoj govora učenika na njemačkom jeziku korištenjem dramapedagoških tehnika. U skladu s obilježjima i karakteristikama akcijskog istraživanja, osmišljen je jedan nastavni sat njemačkog jezika kroz dramapedagoški pristup, u trajanju od 45 minuta. Navedeni je sat ostvaren s učenicima drugog razreda srednje škole. Rezultati istraživanja pokazali su kako su dramapedagoške aktivnosti učenicima zabavne te da promiču njihovu interakciju i motivaciju za aktivnim sudjelovanjem i govorom u nastavi stranog jezika. Učenici su se našli u situacijama koje nisu zahtijevale primjenu samo njihovih intelektualnih vještina, već i govor tijela, zajedničko konstruiranje značenja te emocionalno razumijevanje. Ostvarivanjem dramapedagoškog pristupa u nastavi učenje stranog jezika podiže se na višu razinu jer se, između ostaloga, razvija i učenikova mašta, kreativnost, spontanost i originalnost. Prema tome može se zaključiti kako dramapedagoški pristup nastavi ima puno više prednosti u odnosu na konvencionalnu, tradicionalnu nastavu koja je usmjerena samo na sadržaj i činjenično znanje

PET/CT imaging of spinal inflammation and microcalcification in patients with low back pain: A pilot study on the quantification by artificial intelligence-based segmentation

Background: Current imaging modalities are often incapable of identifying nociceptive sources of low back pain (LBP). We aimed to characterize these by means of positron emission tomography/computed tomography (PET/CT) of the lumbar spine region applying tracers 18F-fluorodeoxyglucose (FDG) and 18F-sodium fluoride (NaF) targeting inflammation and active microcalcification, respectively. Methods: Using artificial intelligence (AI)-based quantification, we compared PET findings in two sex- and age-matched groups, a case group of seven males and five females, mean age 45 \ub1 14 years, with ongoing LBP and a similar control group of 12 pain-free individuals. PET/CT scans were segmented into three distinct volumes of interest (VOIs): lumbar vertebral bodies, facet joints and intervertebral discs. Maximum, mean and total standardized uptake values (SUVmax, SUVmean and SUVtotal) for FDG and NaF uptake in the 3 VOIs were measured and compared between groups. Holm–Bonferroni correction was applied to adjust for multiple testing. Results: FDG uptake was slightly higher in most locations of the LBP group including higher SUVmean in the intervertebral discs (0.96 \ub1 0.34 vs. 0.69 \ub1 0.15). All NaF uptake values were higher in cases, including higher SUVmax in the intervertebral discs (11.63 \ub1 3.29 vs. 9.45 \ub1 1.32) and facet joints (14.98 \ub1 6.55 vs. 10.60 \ub1 2.97). Conclusion: Observed intergroup differences suggest acute inflammation and microcalcification as possible nociceptive causes of LBP. AI-based quantification of relevant lumbar VOIs in PET/CT scans of LBP patients and controls appears to be feasible. These promising, early findings warrant further investigation and confirmation

Allele-specific transcriptional elongation regulates monoallelic expression of the IGF2BP1 gene

<p>Abstract</p> <p>Background</p> <p>Random monoallelic expression contributes to phenotypic variation of cells and organisms. However, the epigenetic mechanisms by which individual alleles are randomly selected for expression are not known. Taking cues from chromatin signatures at imprinted gene loci such as the insulin-like growth factor 2 gene 2 (<it>IGF2</it>), we evaluated the contribution of CTCF, a zinc finger protein required for parent-of-origin-specific expression of the <it>IGF2 </it>gene, as well as a role for allele-specific association with DNA methylation, histone modification and RNA polymerase II.</p> <p>Results</p> <p>Using array-based chromatin immunoprecipitation, we identified 293 genomic loci that are associated with both CTCF and histone H3 trimethylated at lysine 9 (H3K9me3). A comparison of their genomic positions with those of previously published monoallelically expressed genes revealed no significant overlap between allele-specifically expressed genes and colocalized CTCF/H3K9me3. To analyze the contributions of CTCF and H3K9me3 to gene regulation in more detail, we focused on the monoallelically expressed <it>IGF2BP1 </it>gene. <it>In vitro </it>binding assays using the CTCF target motif at the <it>IGF2BP1 </it>gene, as well as allele-specific analysis of cytosine methylation and CTCF binding, revealed that CTCF does not regulate mono- or biallelic <it>IGF2BP1 </it>expression. Surprisingly, we found that RNA polymerase II is detected on both the maternal and paternal alleles in B lymphoblasts that express <it>IGF2BP1 </it>primarily from one allele. Thus, allele-specific control of RNA polymerase II elongation regulates the allelic bias of <it>IGF2BP1 </it>gene expression.</p> <p>Conclusions</p> <p>Colocalization of CTCF and H3K9me3 does not represent a reliable chromatin signature indicative of monoallelic expression. Moreover, association of individual alleles with both active (H3K4me3) and silent (H3K27me3) chromatin modifications (allelic bivalent chromatin) or with RNA polymerase II also fails to identify monoallelically expressed gene loci. The selection of individual alleles for expression occurs in part during transcription elongation.</p

Development and validation of eco-friendly strategies based on thin film microextraction for water analysis

The final publication is available at Elsevier via https://dx.doi.org/10.1016/j.chroma.2018.10.026 © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/The aim of the current study is the establishment of Green Analytical Chemistry strategies for water analysis by elimination/reduction of hazardous chemicals, energy consumption, and waste generation throughout the entire analytical workflow. The first approach introduced in this manuscript consists of addition of water to a sampling vessel that contains a thin film microextraction (TFME) device, followed by removal of the device after equilibration, and subsequent quantification of the extracted components by thermal desorption GC/MS. In this approach, namely the in-bottle TFME approach, analyte-loss associated errors that stem from analyte adherence to glass surfaces and/or degradation are avoided as extraction occurs in situ, while analytes are isolated from a complex matrix that contains degradation agents (bacteria, oxidizing or reducing species, etc.). This procedure also circumvents the splitting of original samples into sub-samples. The second approach involves the use of portable TFME devices that facilitate on-site extraction of compounds, therefore eliminating the use of collection vessels, a factor known to potentially introduce errors into analysis. The herein described procedure involves attachment of the TFME device to drill accessories, analyte extraction via direct immersion into sampled site, and subsequent on-site instrumental analysis, which is carried out with the use of a portable GC/MS containing an appropriate thermal desorption interface, or alternatively, by transferring the membrane to the laboratory for bench-top GC/MS analysis. To facilitate a better understanding of the processes governing the developed approaches, modeling by COMSOL Multiphysics® software was performed. The findings of this study were applied for further method optimization, and the optimized developed methods were then applied for on-site surface water analyses. Finally, the greenness of the developed methods was evaluated with use of the eco-scale assessment, with obtained scores compared to that of the US EPA 8270 method.Ontario Research FundMaxxam Analytics (Mississagua, ON