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

Evaluation Of Agronomic Traits And Adaptation Of Some Promising Salt Tolerant Rice Lines/Varieties Growing In The Coastal Areas In Thanh Hoa Province, Vietnam

Rice (Oryza sativa L.) is the main food in Vietnam and plays an important role in economic activity in this country. However, rice yield and growing areas are adversely influenced by the threats of the devastation caused by the rise of sea level. This study aimed to evaluate the salt effects in field experiments of 19 promising salt tolerance rice lines/varieties carrying QTL/Saltol, which were selected from the BC2F5 breeding combination between FL479 and Bac Thom 7 (BC7). The field experiments were conducted in two consecutive seasons at three saline-affected rice growing areas, including Nga Son, Hoang Hoa and Quang Xuong districts, Thanh Hoa province. The results showed that among those promising rice lines, the HL15 line had the best yield components in all experimented sites, and good agronomic traits with short plant height and shorter growth duration than the BC7 variety (115-117 days) in the spring season and 102-103 days in the summer season, respectively. All tested had the same level of pest resistance/infection as the control variety BT7 under production conditions using pesticides including HL1, HL13 and HL15, which were mildly infected with the bacterial leaf blight, while HL15 variety showed high resistance to pests and diseases. Moreover, the HL2, HL15 and HL19 lines had scored 3, equivalent to FL478 (tolerant), and the survival rate was over 85% compared with the control variety. Overall, our findings showed that the promising purebred variety HL15 had a range of good agronomic characteristics, resistance to major pests and diseases, high yield potential and salinity tolerance of 6‰. The HL15 variety was renamed as SHPT15 and continued to conduct procedures to recognize it as a new rice variety

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)

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

Migration-related emotional distress among Vietnamese psychiatric patients in Germany: An interdisciplinary, mixed methods study

Culture and socialization influence how individuals perceive and express emotional distress. Research therefore, must consider the context to capture individual experiences. However, the majority of studies on factors associated with emotional distress among migrants use quantitative approaches, limiting an in-depth understanding. This study investigates emic themes of emotional distress among Vietnamese migrants by integrating anthropological and psychiatric approaches. The mixed methods study first quantified differences in reported themes of distress between Vietnamese (n = 104) and German (n = 104) patients, who utilized two psychiatric outpatient clinics in Berlin, Germany. Based on these differences, ethnographic interviews were conducted with 20 Vietnamese patients. In the quantitative part, differences in frequency of reported distress between Vietnamese and German patients indicate cultural and migration-related issues among Vietnamese migrants, such as the upbringing of children in a transcultural context. In the qualitative part, interviews with Vietnamese patients elicited contextualizing information and additional themes of distress. Besides commonly expressed socioeconomic themes, such as work and finances, we identified affectively charged themes concerning roles toward partnership and children. A central emic theme is expressed as “moments of speechlessness,” which go beyond a lack of language proficiency and challenge patients in different spheres of life. Migration entails complex affective dynamics, determined by a specific migratory and post-migratory context. Within this context, norms and values determine which themes of distress patients articulate openly. Therefore, an interdisciplinary, mixed-methods approach can yield a contextualized understanding of emotional distress and the complex nature of migration

Severe malaria not responsive to artemisinin derivatives in man returning from Angola to Vietnam.

Resistance to artemisinin derivatives, the most potent antimalarial drugs currently used, has emerged in Southeast Asia and threatens to spread to Africa. We report a case of malaria in a man who returned to Vietnam after 3 years in Angola that did not respond to intravenous artesunate and clindamycin or an oral artemisinin-based combination

Genotyping and antibiotic resistance of thermophilic Campylobacter isolated from chicken and pig meat in Vietnam

Background Campylobacter species are recognized as the most common cause of foodborne bacterial gastroenteritis in humans. In this study nine Campylobacter strains isolated from chicken meat and pork in Hanoi, Vietnam, were characterized using molecular methods and tested for antibiotic resistance. Results The nine isolates (eight C. jejuni and one C. coli) were identified by multiplex PCR, and tested for the presence or absence of 29 gene loci associated with virulence, lipooligosaccharide (LOS) biosynthesis and further functions. flaA typing, multilocus sequence typing and microarray assay investigation showed a high degree of genetic diversity among these isolates. In all isolates motility genes (flaA, flaB, flhA, fliM), colonization associated genes (cadF, docB), toxin production genes (cdtA, cdtB, secD, secF), and the LOS biosynthesis gene pglB were detected. Eight gene loci (fliY, virB11, Cje1278, Cj1434c, Cj1138, Cj1438c, Cj1440c, Cj1136) could not be detected by PCR. A differing presence of the gene loci ciaB (22.2 %), Cje1280 (77.8 %), docC (66.7 %), and cgtB (55.6 %) was found. iamA, cdtC, and the type 6 secretion system were present in all C. jejuni isolates but not in C. coli. flaA typing resulted in five different genotypes within C. jejuni, MLST classified the isolates into seven sequence types (ST-5155, ST-6736, ST-2837, ST-4395, ST-5799, ST-4099 and ST-860). The microarray assay analysis showed a high genetic diversity within Vietnamese Campylobacter isolates which resulted in eight different types for C. jejuni. Antibiotic susceptibility profiles showed that all isolates were sensitive to gentamicin and most isolates (88.8 %) were sensitive to chloramphenicol, erythromycin and streptomycin. Resistance rates to nalidixic acid, tetracycline and ciprofloxacin were 88.9, 77.8 and 66.7 %, respectively. Conclusions To the best of our knowledge, this study is the first report that shows high genetic diversity and remarkable antibiotic resistance of Campylobacter strains isolated from meat in Vietnam which can be considered of high public health significance. These preliminary data show that large scale screenings are justified to assess the relevance of Campylobacter infections on human health in Vietnam