Specific binding of the methyl binding domain protein 2 at the BRCA1-NBR2 locus

The methyl-CpG binding domain (MBD) proteins are key molecules in the interpretation of DNA methylation signals leading to gene silencing. We investigated their binding specificity at the constitutively methylated region of a CpG island containing the bidirectional promoter of the Breast cancer predisposition gene 1, BRCA1, and the Near BRCA1 2 (NBR2) gene. In HeLa cells, quantitative chromatin immunoprecipitation assays indicated that MBD2 is associated with the methylated region, while MeCP2 and MBD1 were not detected at this locus. MBD2 depletion (∼90%), mediated by a transgene expressing a small interfering RNA (siRNA), did not induce MeCP2 or MBD1 binding at the methylated area. Furthermore, the lack of MBD2 at the BRCA1-NBR2 CpG island is associated with an elevated level of NBR2 transcripts and with a significant reduction of induced-DNA-hypomethylation response. In MBD2 knockdown cells, transient expression of a Mbd2 cDNA, refractory to siRNA-mediated decay, shifted down the NBR2 mRNA level to that observed in unmodified HeLa cells. Variations in MBD2 levels did not affect BRCA1 expression despite its stimulation by DNA hypomethylation. Collectively, our data indicate that MBD2 has specific targets and its presence at these targets is indispensable for gene repression

Analysis of Polyphenols from Laminaria hyperborea for application as Life Science ingredients

Makroalger, også kjent som tang og tare, er anerkjent som en verdifull kilde til bioaktive forbindelser. Av disse har polyfenoler fått særlig oppmerksomhet. Polyfenoler har blitt assosiert med en rekke helsefordeler, og er mest kjent for sine ulike biologiske egenskaper, så som antioksidant-, antiinflammatorisk-, antidiabetisk-, og antikreftegenskaper. og. Dette prosjektet undersøkte to typer makroalger; den brune algen Laminaria hyperborea (nor.: stortare) og grønnalgen Ulva intestinalis (nor.: tarmgrønske). De første ekstraksjonene av L. hyperborea blader var utfordrerne grunnet dere høye polysakkarid innhold. Derfor ble det utført en innledende studie med grønn algen U. intestinalis for å utforske de ulike analyseteknikkene som brukes for kvantifisering og identifisering av polyfenoler. Teknikkene som ble benyttet i denne studien inkluderte tradisjonell DAD-HPLC, en totalt-fenolinnhold (TPC) test med Folin-Ciocalteu reagens, en totalt-flavonoidinnhold (TFC) test med en AlCl3 reagens, samt 1H kvantitativ NMR (qNMR) – inkludert 2D NMR. Resultatene viste at det polyfenoliske innholdet var komplekst, og med lav konsentrasjon. For råprøven (crude) av U. intestinalis ble det totale fenolinnholdet målt til 11.3 ± 1.4 mg GAE/g ved bruk av HPLC, 5.0 ± 1.0 mg GAE/g med TPC, og 27.3 ± 2.7 mg GAE/g med qNMR. TFC-testen oppdaget imidlertid ingen flavonoider i råprøven. Denne innledende studien avdekket et sterkt behov for å optimalisere de analytiske metodene som ble benyttet. I tillegg ble det utført LC-MS karakterisering på opprensede prøver av U. intestinalis. Disse identifiserte tentativt tilstedeværelsen av flere fenolsyrer og flavonoider. Noen av disse ble også bekreftet av referansestandarder. Etter optimalisering av TPC-testen og 1H qNMR-metoden, ble kvantifisering og sammenlikning av L. hyperborea med fire andre brunalger gjennomført. Det totale polyfenolinnholdet i L. hyperborea ble funnet å være 5.51 ± 0.00 mg GAE/g (qNMR) og 5.72 ± 0.07 mg GAE/g (TPC). For tang som tilhører Laminariaceae-familien, som vokser i sublitoralsone, viste TPC-testen og selektiv qNMR liknende, men lave polyfenolkonsentrasjoner. Samtidig ble det for tang som vokser i epilittoralsone, som Ascophyllum nodosum (nor.: grisetang) og Fucus vesiculosus (no.: blæretang), observert en større forskjell mellom TPC og qNMR resultatet. Tang som vokser i strand sonen vil bli utsatt for mer lys, noe som kan føre til at det polyfenolske innholdet består av flere polyfenoler som inneholder flere reagerende grupper enn referanse standarden (gallesyre eller floroglucinol) som benyttes, noe som kan produsere overestimering av polyfenol innholdet ved bruk at TPC testen. Det er også mulig at disse artene som vokser nærmere havoverflaten inneholder en økt mengden TPC-forstyrrende forbindelser enn artene som vokser dypere i den sublitorale sonen. For å karakterisere det polyfenoliske innholdet i L. hyperborea, ble flash-kromatografi og preparativ HPLC benyttet til å rense opp tangprøven, for å opparbeide rensede fraksjoner til bruk i analysen. Resultatene viste at prøver som var mer opprenset korresponderte med høyere polyfenolinnhold både i TPC-testen og ved selektiv qNMR. I tillegg viste ORAC-testresultatene økende antioksidantaktivitet med økende renhet. For å identifisere den molekylære strukturen til polyfenolene i L. hyperborea, ble både lavt- og høytoppløselig LC-MS benyttet. Analysene avdekket at polyfenolinnholdet hovedsakelig var sammensatt av lavmolekylære forbindelser, der 96% av de tentativt identifiserte forbindelsene hadde masser under 800 Da. 11 polyfenoliske forbindelser ble identifisert, inkludert fenolsyrer og florotanniner, og flere ble funnet å være sulfaterte, hvilket antas å skyldes en økologisk tilpasning til det marine miljøet. Dette prosjektet gir den først omfattende beskrivelsen av stortares polyfenoliske innhold. Resultatene gir en grundig identifisering av polyfenolinnholdet i bladene av L. hyperborea og antyder potensialet for en «total utnyttelses»-strategi innen kommersiell alginatproduksjon. Videre gir prosjektet verdifull molekylær innsikt i det fenoliske innholdet i tang, noe som gir viktig informasjon til videre forskning på tang- bioraffineriet, kjemisk økologi, og havovervåkning.Macroalgae, or seaweed, has been recognized as a valuable source of bioactive compounds. Among these, polyphenols have gained particular attention. Polyphenols are well-known for their various bioactivities, such as antioxidant, anti-inflammatory, antidiabetic, and anticancer activities, and have been associated with a range of health benefits. This project focused on the investigation of two types of macroalgae, namely the leaf biomass of the brown algae Laminaria hyperborea (eng.: tangle/cuvie) and the green algae Ulva intestinalis (eng.: gut weed). The initial extractions of L. hyperborea leaves were challenging due to its high polysaccharide content. Therefore, an initial case study was conducted on U. intestinalis to explore the combination of analytical techniques employed for polyphenol quantification and identification in seaweeds. The techniques utilized in this investigation included DAD-HPLC, the total phenolic content (TPC) assay with the Folin–Ciocalteu (FC) reagent, total flavonoid content (TFC) assay with an AlCl3 reagent, and 1H qNMR – including 2D NMR. The analyses showed a relatively composed polyphenolic nature with lower individual concentrations. For the crude sample of U. intestinalis, the total phenolic content was found to be 11.3 ± 1.4 mg GAE/g using HPLC, 5.0 ± 1.0 mg GAE/g with TPC, and 27.3 ± 2.7 mg GAE/g with qNMR. However, the TFC assay detected no flavonoids in the crude sample. The case study strongly indicated the need for optimization of the analytical methods. In addition, characterization using LC-MS was performed on purified samples of U. intestinalis, which tentatively identified the presence of several phenolic acids and flavonoid aglycones. Some of these were confirmed using reference standards. Following optimization of the TPC assay and the 1H qNMR method, the quantification and comparison of Laminaria hyperborea to four other brown algae was conducted. The total polyphenolic content of L. hyperborea was found to be 5.51 ± 0.00 mg GAE/g (qNMR) and 5.72 ± 0.07 mg GAE/g (TPC). For seaweeds belonging to the Laminariaceae family, which grow in the sublittoral zone, the TPC assay and selective qNMR showed similar, lower polyphenol yields. Still, a larger difference was observed between the TPC and qNMR results for seaweed growing in the eulittoral zone, such as Ascophyllum nodosum (eng.: knotted kelp) and Fucus vesiculosus (eng.: bladder wrack). Seaweed growing in more shallow waters will have higher light accessibility, possibly producing a polyphenolic pool with an increased portion of polyphenols containing an increased number of reacting groups compared to the reference standard (gallic acid or phloroglucinol) used, thus resulting in an overestimation when quantifying with the TPC assay. It is also possible that these species in more shallow waters have a higher occurrence of TPC-interfering compounds than seaweed species growing in the sublittoral zone. To characterize the polyphenolic content of L. hyperborea, flash chromatography and preparative HPLC were used to purify the seaweed sample, obtaining purified fractions for analysis. Results showed that increasing purification corresponded to higher polyphenolic content, as determined by both the TPC assay and selective qNMR. In addition, ORAC assay results revealed increasing antioxidant activity with increasing purification. To identify the molecular structure of polyphenols in L. hyperborea, both low- and high-resolution LC-MS were utilized. Analysis showed that the polyphenolic matrix was mostly composed of low-molecular weight polyphenols, with 96% of the tentatively identified compounds having masses below 800 Da. Eleven polyphenolic compounds were confirmed, including phenolic acids and phlorotannins, and several were found to be sulfated, which is believed to be an ecological adaptation to the marine environment. This project provides the first comprehensive characterization of the polyphenolic content of L. hyperborea leaves. These results provide a characterization of the polyphenolic content of L. hyperborea’s leaf biomass, important for the implementation of a “total utilization” strategy in commercial alginate production. Moreover, the project provides valuable molecular-level insights into the phenolic content of seaweed, yielding valuable implications for research across disciplines, such as in the seaweed biorefinery, chemical ecology, and ocean monitoring.Doktorgradsavhandlin

to the sides of this body

This is a written reflection on the performance and process of making "to the sides of this body" together with Marlene Bonnesen during our MA in Dance

Towards a prototype paleo-detector for supernova neutrino and dark matter detection

Using ancient minerals as paleo-detectors is a proposed experimental technique expected to transform supernova neutrino and dark matter detection. In this technique, minerals are processed and closely analyzed for nanometer scale damage track remnants from nuclear recoils caused by supernova neutrinos and possibly dark matter. These damage tracks present the opportunity to directly detect and characterize the core-collapse supernova rate of the Milky Way Galaxy as well as the presence of dark matter. Current literature presents theoretical estimates for these potential tracks, however, there is little research investigating the experimental feasibility of this technique. At the University of North Florida, we have contributed to the field by searching for and analyzing these damage tracks in prototype detectors constructed from selected minerals, including: halite, Muscovite mica, and Phlogopite mica. This research seeks to characterize the applicable backgrounds in these prototype detectors. We have employed non-destructive techniques, including laser confocal and atomic force microscopy to identify and characterize damage tracks in the minerals. Chemical etching and plasma etching of target minerals is used to enhance the detectability of these damage tracks at the expense of altering some of their geometrical attributes. With the use of an etching rate model and automatic track detection via Python, damage track lengths will be reconstructed. Our data is compared to current theoretical predictions to pursue the practical implementation of paleo-detectors as local core-collapse supernova neutrino and dark matter detectors

Au nom du Bon Dieu et du Buffalo: Metis Lived Catholicism on the Northern Plains

This dissertation argues that Metis lived Catholicism was a tool of identity formation, resistance to colonialism, and political action among bison hunters of the northern plains in the long nineteenth century. The Catholicity of Metis bison hunters quotidian is highlighted through the extensive Michif French written legacy of Turtle Mountain historian ChWeUm (William Jr.) Davis (1845-1937). Daviss biography anchors a Metis national memory, weaving stories and events from both sides of the Medicine Line. His life story and the religious experiences of his relatives come together to explain why some Metis people adhered to Catholicism and its practices. Sustained experiences of the divine helped Metis families adapt and resist the effects of settler colonialism on the northern plains, including the end of organized bison-hunting expeditions. This dissertation blends several methodologies social history, biography, ethnohistory, and social network analysis from the digital humanities to interrogate the history of Catholicism among Metis peoples

Factory management

Thesis (M.B.A.)--Boston University, 1939, p.19 missing in origina

Shaping Service Delivery For Cervical Cancer Screening: Understanding Knowledge, Acceptability And Preferences Among Women In The Neno District Of Malawi

Background: Located in south-eastern Africa, Malawi is a low income country with a population of 16.4 million. It is currently estimated to have the highest age-standardized rate of cervical cancer in the world at 75.9 per 100,000 population. Cervical cancer accounts for 28% of all reported female cancers in the country, and 80% of women diagnosed with the disease die from it, making cervical cancer a public health priority for the country. Despite the fact that cervical cancer screening services (by visual inspection with acetic acid (VIA)) has been offered free of charge at public health facilities in the country since 2004, the services remain underutilized across the country. The purpose of this study was to explore potential factors associated with the acceptability and utilization of cervical cancer screening services among adult women in the Neno district of Malawi, and to identify their service delivery preferences in order to guide the design of effective cervical cancer education and screening programs. Methods: This study employed an exploratory qualitative research study design. Data was collected between July and August 2015 in the district of Neno, Malawi by conducting in-depth semi-structured interviews within a sample of 50 women between the ages of 18-55 years. Participants were recruited among women attending health facilities and community screening events within the district using a purposive sampling technique. Constructs from the Health Belief Model and Socio-Ecological Model were used to construct a conceptual framework for this study. Results: Several themes emerged from the data, namely: misconceptions about how cervical cancer develops, low perceptions of risk of the disease, effects of interpersonal interactions on preventative behaviors, barriers limiting access to cervical cancer screening services, and service preferences for the integration of cervical cancer screening. Despite limited understanding about the transmission and development of cervical cancer, knowledge about its risk factors and signs and symptoms was relatively high among the study sample. Perceived barriers to using cervical cancer prevention services included: distance from health facilities, challenges in accessing transportation, limited availability and awareness of services, logistical factors, and a low perception of risk. Facilitators to using the services were based on: high satisfaction with healthcare, strong desires to know about one’s health status, interpersonal interactions with health professionals, husband’s approval, and faith in the curative abilities of western medicine. Women in the sample reported high acceptability and intention to obtain screening if the services were available at a nearby location. The study sample was also highly receptive of integrated approaches to receiving cervical cancer screening both at health facilities and community screening events Conclusions: Knowledge and awareness about cervical cancer and its risk factors among the study sample was relatively high, indicating that sporadic health education and awareness raising about the disease has been taking place throughout health facilities within the Neno district. Misconceptions about the disease however highlight the importance of having a systematic method and coordinated strategy for delivering cervical cancer health education. The expansion of cervical cancer screening services across all health centers in the district is needed in order to address barriers to accessing preventative services and to increase coverage and utilization rates for VIA screening services in Neno

Meiotic Maturation of the Mouse Oocyte Requires an Equilibrium between Cyclin B Synthesis and Degradation

AbstractAmong the proteins whose synthesis and/or degradation is necessary for a proper progression through meiotic maturation, cyclin B appears to be one of the most important. Here, we attempted to modulate the level of cyclin B1 and B2 synthesis during meiotic maturation of the mouse oocyte. We used cyclin B1 or B2 mRNAs with poly(A) tails of different sizes and cyclin B1 or B2 antisense RNAs. Oocytes microinjected with cyclin B1 mRNA showed two phenotypes: most were blocked in MI, while the others extruded the first polar body in advance when compared to controls. Moreover, these effects were correlated with the length of the poly(A) tail. Thus it seems that the rate of cyclin B1 translation controls the timing of the first meiotic M phase and the transition to anaphase I. Moreover, overexpression of cyclin B1 or B2 was able to bypass the dbcAMP-induced germinal vesicle block, but only the cyclin B1 mRNA-microinjected oocytes did not extrude their first polar body. Oocytes injected with the cyclin B1 antisense progressed through the first meiotic M phase but extruded the first polar body in advance and were unable to enter metaphase II. This suggested that inhibition of cyclin B1 synthesis only took place at the end of the first meiotic M phase, most likely because the cyclin B1 mRNA was protected. The injection of cyclin B2 antisense RNA had no effect. The life observation of the synthesis and degradation of a cyclin B1–GFP chimera during meiotic maturation of the mouse oocyte demonstrated that degradation can only occur during a given period of time once it has started. Taken together, our data demonstrate that the rates of cyclin B synthesis and degradation determine the timing of the major events taking place during meiotic maturation of the mouse oocyte