Towards site-specific functional analysis of RNA N6-methyladenosine and 5-methylcytosine in Arabidopsis thaliana

Phenotype prediction from DNA or RNA sequence in eukaryotes is difficult as a result of the multiple layers of gene expression regulation. One of these regulatory layers is RNA modification which occurs either co-transcriptionally or post-transcriptionally to RNA and affects many aspects of RNA biology. Recent transcriptome-wide insights of RNA modifications have begun to elucidate the extent of this landscape, leading to the proposition of the “epitranscriptome”. However, understanding the “epitranscriptome” and its consequences to RNA metabolism, functional relevance and mechanism of action remains an enormous undertaking. The lack of tools to specifically manipulate an RNA modification has hampered the un-biased evaluation of their importance as well as understanding of the mechanism underlying their activities in a site-specific manner. During my research, I investigated the RNA-guided RNA targeting system CRISPRdCas13 and the RNA-based technique Short Tandem Target Mimic (STTM) regarding their potential to develop programmable systems for targeted demethylation of two RNA modifications N6-methyladenosine (m6A) and 5-methylcytosine (m5C) in Arabidopsis thaliana. Initial results indicated unsuccessful interference to m5C deposition using the STTM approach. In contrast, fusion of dCas13 to RNA modifying domains ALKBH10B or human TEN-ELEVEN TRANSLOCATION1 (hTET1) respectively enabled alterations of m6A- or m5C-bearing reporter or endogenous transcripts in either Nicotiana benthamiana or A. thaliana. However, further investigation is required to understand the robustness of both approaches. Possible improvements are discussed for targeted RNA demethylation tools using CRISPRdCas13. In addition, the functional relevance of RNA m5C in root development in A. thaliana was also extensively investigated with regards to previously proposed involvement of two transcripts SHORT HYPOCOTYL 2 (SHY2/IAA3) and INDOLEACETIC ACID-INDUCED PROTEIN 16 (IAA16). Phenotypic and mRNA quantification analyses of m5C-deficient and SHY2/IAA3 loss-of-function mutants refuted a key role for SHY2/IAA3 and IAA16 in m5C regulation of root development. Although I identified an m5C site C348 on SHY2/IAA3 using bisulfite RNA amplicon sequencing and initial nucleotide conservation analysis suggested that this site is more conserved than other non-methylated cytosines within SHY2/IAA3 mRNA, functional analysis using dCas13-hTET1 conjugate did not support the importance of this C348 in planta. In summary, RNA-guided CRISPR-dCas13 and multi-strategy approaches for site-specific functional study offers potential to significantly improve our understanding of RNA modifications.Thesis (Ph.D.) -- University of Adelaide, School of Biological Sciences, 202

Teacher returnees from overseas programs in the west: a narrative study in Vietnam

In this narrative inquiry research, our focus was on exploring teacher identity within the context of the expanding field of language teacher education and professional development. Utilizing theories of teacher identities to analyze data from interviews and reflections, we observed that conversations and stories were frequently shared in social settings, with overseas teaching practices being a substantial influence on their reflections. The findings indicated that the participants' teacher identities continually changed and dynamically evolved, particularly influenced by their engagement in overseas educational programs. Upon their return, participants expressed hope in maintaining their newly transformed perspectives on teaching and learning. In conclusion, we discussed further research directions and implications to underscore the ongoing significance of this topic

Purification and characterization of novel fibrinolytic proteases as potential antithrombotic agents from earthworm Perionyx excavatus

Six protease fractions, namely FI, FII, FIII-1, FIII-2, FIII-3 and FIV, were isolated from Perionyx excavatus earthworm biomass by acetone precipitation, followed by serial chromatography using anion exchange, hydrophobic interaction and size exclusion chromatography. All fractions exhibited strong hydrolytic activity towards casein. The activity of six fractions towards fibrin, determined by fibrin plate assay, ranged from 44 to 831 plasmin unit.mg-1 and ranked as FIII-3 > FIII-2 > FI > FIII-1 > FIV > FII. Casein degradation was optimal at pH 7 and 11, and at 45-60°C. All fractions were considerably stable at high temperature and wide pH range. They were completely inhibited by phenylmethylsulfonyl fluoride (PMSF). The molecular weights (MW) and isoelectric points (pI) determined by 2D-electrophoresis were 27.5-34.5 kDa, and 4.3-5.2, respectively. Tandem mass spectrometry (MS) analysis was used to deduce the amino acid sequences of some peptides from FIII-1 and FIII-2. The sequences shared 16.9% and 13.2% similarity, respectively, with the fibrinolytic enzymes from two related earthworm species, Lumbricus rubellus and Eisenia fetida. The P. excavatus proteases were classified as serine proteases. They could perform rapid hydrolysis on both coagulated fibrous fibrin and soluble fibrinogen monomers without the presence of activators such as tPA or urokinase

Globalization And Its Effect On Post-Millennium Literature

Globalization, as a slowly spreading risk factor, covers almost the whole country with its full force. Literature and literary studies have evolved into a space for the evocation, promotion and interpretation of various social, political, literary and cultural ideas in the world of globalization. Indian Literature after Independence of India has seen some significant changes in the literary writings. Today, the younger generation in the country is searching for a new identity and they are at the intersection of Individuality, Culture and Society. In the world of literature, the call-center generation sees itself as India’s cultural commissars, projecting India as a land of illumination and emancipation. The major Samson of the youth lies in the power of the youth through the unlimited consumption of natural and human resources. The novels of the millennium assist us in dispelling the myth that contemporary writing is only for the light entertainment of the younger generations. &nbsp

Artikulationen des Affektiven und die Formierung transkultureller Emotionsrepertoires im Vietnamesischen Berlin

Aus anthropologisch-psychiatrischer Perspektive thematisiert unser Beitrag die Formierung transkultureller Emotionsrepertoires in den Lebensentwürfen vietnamesischer Migrant_innen der ersten Generation in Berlin. Konkret gilt unser empirisches Interesse den affektiven Anstrengungen von Migration, die sich im Leben von vietnamesischen Patient_innen abzeichnen, die psychiatrisch- psychotherapeutische Hilfe in Anspruch nehmen, sowie von deren Angehörigen. Zum einen möchten wir der Frage nachgehen, wann genau affektive Krisenerfahrungen zu Belastungen werden, die auf Basis bisheriger Emotionsrepertoires nicht länger zu bewältigen sind und zu einer Inanspruchnahme psychiatrisch-psychotherapeutischer Hilfe führen. Zum anderen möchten wir Antworten auf die Frage geben, inwiefern diese Inanspruchnahme zu einer Herausbildung neuer und dezidiert transkultureller Emotionsrepertoires beiträgt, die Beheimatungsprozesse begünstigen können, welche in neue und multiple Zugehörigkeiten und/oder Nichtzugehörigkeiten münden (Pfaff-Czarnecka 2012, Scheer 2014, Lähdesmäki et al. 2016, Röttger-Rössler 2016). Die Relevanz der letzteren Frage spiegelt sich in der zunehmenden Akzeptanz vietnamesisch- sprachiger und transkulturell sensibler psychiatrischer Versorgungsan-gebote im Rahmen der Eröffnung zweier Spezialambulanzen in Berlin wider (Ta et al. 2015,Hahn et al. 2016). Diese Versorgungsangebote sind eingebettet in ein aktives Netzwerk, welches die Förderung der seelischen Gesundheit von vietnamesischen Migrant_innen in Deutschland dient und dabei Träger sozialer Hilfen miteinander in Beziehung setzt und auch transnationale psychiatrische Perspektiven miteinbezieht (Ta et al. 2016b). Aus unseren ethnographischen Begegnungen, Beobachtungen und Gesprächen geht hervor, dass das erwähnte Setting in den Spezialambulanzen für unsere Gesprächspartner_innen einen einzigartigen Artikulationsraum darstellt, in dem in bislang ungewohnter Weise und jenseits von Stigmatisierungsängsten über affektive Erfahrungen, Anstrengungen und Belastungen gesprochen werden kann. Wer sich in Vietnam in psychiatrische Behandlung begebe oder in eine psychiatrische Klinik „eingewiesen“ werde, habe es laut unserer Gesprächspartner_innen nicht nur in Bezug auf die Qualität der Versorgung schlechter als in Deutschland: Der-/diejenige werde schnell als điên bezeichnet und müsse mit Diskriminierung und Stigmatisierung in Form eines Gesichtsverlusts rechnen, der sich auch auf das familiäre Umfeld ausweite. Das Wort điên bedeute „verrückt sein“, umgangssprachlich steht es für den medizinischen Ausdruck „an einer psychischen Krankheit leiden“ (bị bệnh tâm thần). Jedoch birgt es durchweg negative Konnotationen in sich, da Menschen, die als điên bezeichnet werden, nicht länger ernst genom-men und aus ihrem sozialen Umfeld ausgegrenzt würden. Affektive Belastungen werden aus diesem Grund häufig verdeckt, um einerseits nicht gegen die Gebote der Wahrung sozialer Harmonie zu verstoßen und andererseits, um eine individuelle und familiäre Stigmatisierung zu vermeiden (Lauber & Rössler 2007, Machleidt 2013). Selbstverständlich gibt es auch in Vietnam vielfältige Bewältigungsstrategien. In Migrationserfahrungen fußende affektive Belastungen beinhalten unserer Meinung nach aber andere Herausforderungen, welche die vertrauten Strategien in dem veränderten sozialen, politischen und wirtschaftlichen Kontext oftmals an ihre Grenzen stoßen lassen. Im Folgenden erläutern wir zunächst, inwiefern wir Affekte von Emotionen im Kontext von Migration konzeptuell unterscheiden und was unter einem transkulturellen Emotionsrepertoire zu verstehen ist. Des Weiteren wird die psychiatrische Ambulanz als ein besonderer Artikulationsraum des Affektiven vorgestellt, sowie auch als Forschungsraum unseres interdisziplinären Projektes. Sodann gewähren wir Einblicke in die Lebensentwürfe von zwei Gesprächspartnern, die beide der ersten Generation vietnamesischer Migrant_innen angehören. Um auch eine transgenerationale Sicht auf Affekte und Emotionen in der Migration zu ermöglichen, beschließen wir unseren empirischen Teil mit einer Beschreibung der Ansichten einer Gesprächspartnerin der zweiten Generation, deren Mutter in psychiatrisch- psychotherapeutischer Be-handlung ist. Unser Beitrag endet mit einer Diskussion, in der wir gängigen Vorstellungen widersprechen, die die Inanspruchnahme psychiatrisch-psychotherapeutischer Hilfe als ein Zeichen von Hilflosigkeit werten und insbesondere Patient_innen mit migrationsbezogenem Hintergrund Handlungsmacht und Kompetenz absprechen (Pratt Ewing 2005, Kirmayer 2007)

An Ultra-high Quality Factor Terahertz Photonic Crystal Cavity

oai:ojs.rev-jec.org:article/345High quality factor Terahertz (THz) cavities are highly desired for many THz applications. This paper presents an ultra-high quality factor terahertz planar photonic crystal cavity at 300 GHz range. Two approaches are employed to reduce the losses in the cavity increasing the quality factor of the cavity. Firstly, short embedded photonic crystal waveguides are employed to reduce the in – plane loss. Secondly, a novel way of hole displacement is adopted for four edged holes of the L3 – type photonic crystal cavity to decrease the radiation loss. An ultra – high quality factor of 65000 at a resonant frequency of 315.3 GHz was achieved for the designed cavity. This result could enable promising applications such as THz sensing

Mental Health Determinants Among a Psychiatric Outpatient Sample of Vietnamese Migrants in Germany

Background: Mental health risk-factors for Asian migrants have been studied almost exclusively in the US, Canada, and Australia but not in European countries. Therefore, we aimed to identify sociodemographic, clinical, and migration-surrounding factors associated with experienced mental distress among Vietnamese migrants in Germany. Method: 305 Vietnamese migrants utilizing Germany's first Vietnamese psychiatric outpatient clinic filled out at admission the Brief-Symptom-Inventory 18 (BSI-18) as well as a questionnaire on 22 potential mental health determinants. Using a multiple linear regression model, we identified those sociodemographic, clinical, and migration-surrounding factors that were significantly related to the Global Severity Index (GSI) of the BSI-18. Results: The factors unemployment (B = -6.32, p = 0.014), financial problems (B = -10.71, p < 0.001), no or only little religious involvement (B = -3.23, p = 0.002), no psychiatric precontact (B = -7.35, p = 0.004), previous migration experiences (B = 8.76, p = 0.002), and perceived discrimination (B = 6.58, p = 0.011) were found to significantly increase the level of mental distress according to the BSI-GSI. Conclusion: Based on these results, we were able to construct a mental health risk-profile for Vietnamese migrants in Germany, which aims to detect candidates for psychiatric problems earlier and supply them with customized prevention and therapy options

Lovastatin for the Treatment of Adult Patients With Dengue: A Randomized, Double-Blind, Placebo-Controlled Trial.

BACKGROUND: Dengue endangers billions of people in the tropical world, yet no therapeutic is currently available. In part, the severe manifestations of dengue reflect inflammatory processes affecting the vascular endothelium. In addition to lipid lowering, statins have pleiotropic effects that improve endothelial function, and epidemiological studies suggest that outcomes from a range of acute inflammatory syndromes are improved in patients already on statin therapy. METHODS: Following satisfactory review of a short pilot phase (40 mg lovastatin vs placebo in 30 cases), we performed a randomized, double-blind, placebo-controlled trial of 5 days of 80 mg lovastatin vs placebo in 300 Vietnamese adults with a positive dengue NS1 rapid test presenting within 72 hours of fever onset. The primary outcome was safety. Secondary outcomes included comparisons of disease progression rates, fever clearance times, and measures of plasma viremia and quality of life between the treatment arms. RESULTS: Adverse events occurred with similar frequency in both groups (97/151 [64%] placebo vs 82/149 [55%] lovastatin; P = .13), and were in keeping with the characteristic clinical and laboratory features of acute dengue. We also observed no difference in serious adverse events or any of the secondary outcome measures. CONCLUSIONS: We found lovastatin to be safe and well tolerated in adults with dengue. However, although the study was not powered to address efficacy, we found no evidence of a beneficial effect on any of the clinical manifestations or on dengue viremia. Continuing established statin therapy in patients who develop dengue is safe.Chinese Clinical Trials Registration. ISRCTN03147572

Implementation of point-of-care testing of C-reactive protein concentrations to improve antibiotic targeting in respiratory illness in Vietnamese primary care: a pragmatic cluster-randomised controlled trial

Background In previous trials, point-of-care testing of C-reactive protein (CRP) concentrations safely reduced antibiotic use in non-severe acute respiratory infections in primary care. However, these trials were done in a research-oriented context with close support from research staff, which could have influenced prescribing practices. To better inform the potential for scaling up point-of-care testing of CRP in respiratory infections, we aimed to do a pragmatic trial of the intervention in a routine care setting. Methods We did a pragmatic, cluster-randomised controlled trial at 48 commune health centres in Viet Nam between June 1, 2020, and May 12, 2021. Eligible centres served populations of more than 3000 people, handled 10–40 respiratory infections per week, had licensed prescribers on site, and maintained electronic patient databases. Centres were randomly allocated (1:1) to provide point-of-care CRP testing plus routine care or routine care only. Randomisation was stratified by district and by baseline prescription level (ie, the proportion of patients with suspected acute respiratory infections to whom antibiotics were prescribed in 2019). Eligible patients were aged 1–65 years and visiting the commune health centre for a suspected acute respiratory infection with at least one focal sign or symptom and symptoms lasting less than 7 days. The primary endpoint was the proportion of patients prescribed an antibiotic at first attendance in the intention-to-treat population. The per-protocol analysis included only people who underwent CRP testing. Secondary safety outcomes included time to resolution of symptoms and frequency of hospitalisation. This trial is registered with ClinicalTrials.gov, NCT03855215. Findings 48 commune health centres were enrolled and randomly assigned, 24 to the intervention group (n=18 621 patients) and 24 to the control group (n=21 235). 17 345 (93·1%) patients in the intervention group were prescribed antibiotics, compared with 20 860 (98·2%) in the control group (adjusted relative risk 0·83 [95% CI 0·66–0·93]). Only 2606 (14%) of 18 621 patients in the intervention group underwent CRP testing and were included in the per-protocol analysis. When analyses were restricted to this population, larger reductions in prescribing were noted in the intervention group compared with the control group (adjusted relative risk 0·64 [95% CI 0·60–0·70]). Time to resolution of symptoms (hazard ratio 0·70 [95% CI 0·39–1·27]) and frequency of hospitalisation (nine in the intervention group vs 17 in the control group; adjusted relative risk 0·52 [95% CI 0·23–1·17]) did not differ between groups. Interpretation Use of point-of-care CRP testing efficaciously reduced prescription of antibiotics in patients with non-severe acute respiratory infections in primary health care in Viet Nam without compromising patient recovery. The low uptake of CRP testing suggests that barriers to implementation and compliance need to be addressed before scale-up of the intervention. Funding Australian Government, UK Government, and the Foundation for Innovative New Diagnostics