EFFORTS TO IMPROVE CHILDREN’S LANGUAGE SKILLS THROUGH THE WORD GUESSING METHOD WITH PICTURE CARD MEDIA

This research is based on pre-action observation results, which indicate that children's language skills at RA Tarbiyatul Banin are still low, especially in vocabulary recognition and understanding letter or word concepts. The aim of this study is to improve children's language skills through the word-guessing method assisted by picture card media at RA Tarbiyatul Banin, Plosorejo Village, Pucakwangi District, Pati Regency, in the 2019/2020 academic year. This issue arises because children's language skills in word-guessing activities are not optimal due to monotonous teaching methods and a lack of activity variation, causing children to feel bored and uninterested. Consequently, their cognitive abilities remain underdeveloped. The research subjects consist of 26 children from RA Tarbiyatul Banin in the 2019/2020 academic year, including 13 boys and 13 girls. The research was conducted at RA Tarbiyatul Banin, Plosorejo Village, Pucakwangi District, Pati Regency. This study employs a classroom action research (CAR) approach, which consists of four stages: (1) Planning, (2) Implementation, (3) Observation, and (4) Reflection. Data collection techniques include observation and documentation. The research was carried out in three cycles: pre-action, Cycle 1, and Cycle 2. The results indicatean improvement in children's language skills through the word-guessing method. This is evident from the pre-cycle learning outcomes, where no children were able to fully participate. However, after improvements in learning during Cycle 1, the "Developing as Expected" (BSB) scale increased by 1.92%. In Cycle 2, the "Well Developed" (BSB) scale reached 25%. Based on these percentage results, the researcher concludes that the word-guessing game assisted by picture cards can effectively improve children's language skills at RA Tarbiyatul Banin, Plosorejo Village, Pucakwangi District, Pati Regency, in the 2022/2023 academic year

التعاليم الدينية في رواية العائدة لسلام أحمد إدريسو:دراسة سميائية لجوليا كريستيفا

رواية العائدة لسلام أحمد إدريسو هي الرواية الحافلة بالقيمة الدينية. التعاليم الدينية لها دور مهمّ في عملية تحويل أخلاق المجتمع إلى الجهة الأحسن وتكون هدى للناس. في هذا البحث، استفادت الباحثة دراسة سميائية لكريستيفا في بحث تلك الرواية لأنّ هذه الدراسة أكبر مناسبا لاكتشاف القيمة الدينية المضمونة في تلك الرواية و ترابطها بالنص ما يكون مرجعا لها في القرآن و الحديث. وغرض هذا البحث هو لمعرفة شكل القيمة الدينية المضمونة في رواية العائدة لسلام أحمد إدريسو و ترابطها بالنص ما يكون مرجعا لها في القرآن و الحديث عند نظر دراسة سميائية لكريستيفا. وتعتمد الباحثة على المنهج الوصفي التحليلي في تحليل هذا البحث وهو المنهج المستخدم بطريقة وصف البيانات عن القيمة الدينية المضمونة في رواية العائدة لسلام أحمد إدريسو، ثم عمل التحليل اليها بمساعدة المنهج المقارن يعني قارنت القيم الدينية بنص المرجع من آيات القرآن و الحديث ما تؤيّد المعنى لتلك القيمة الدينية باستفادة دراسة سميائية لكريستيفا. أشكال القيمة الدينية المضمونة في تلك الرواية هي : (١) العقيدة التي تدور على الإيمان، الإيمان بالله و ملائكته و كتبه و رسله و باليوم الآخر و بالقضاء خيره و شرّه من الله تعالى؛ (٢) الأخلاق التي تشتمل على الأخلاق للخالق و الأخلاق للناس و العالَم؛ (٣) الشريعة التي تعمل الناس لتحقيق عن رِضا الله كمثل العبادة، المعاملة، وغير ذلك. يمكن تلك القيمة الدينية الثالثة قد تراجع من آيات القرآن و الحديث لتناصية. بناء على طريقة تناصية لكريستيفا، يعرف ان نص الرواية في تصوير القيمة الدينية قد عمل عملية تدوّل لأنّ النص في الرواية تتدوّل النص في القرآن و الحديث و ينقل المعنى الوريد فيه

ANALISIS DAMPAK GAYA TUMBUKAN MELALUI PERUBAHAN BENTUK SEGI CRUSH INITIATOR PADA STRUKTUR TAILOR-WELDED BLANK

Tailor-welded blank (TWB) structure is one of the vehicle component models used in vehicle parts. In addition, these structures can be used in crashworthiess technology which can reduce injuries during a collision.. The structure of a vehicle that has greater strength can cause passengers to be thrown from the passenger compartment. This paper discusses the effect of faceted holes such as square, hexagonal and octagonal as crush initiator mounted on TWB which results in a maximum impact force (Fmax). The smallest maximum impact force is the criteria achieved from this study. The method used experimental quasi-static loading of the actuator speed of 0.5 mm/s to achieve 9.5 mm of deformation. TWB was made from plate formation with a process of stamping to spot weld. The result showed that the maximum impact force has an increase directly proportional to the addition of the shape of the crush initiator in the amount of 14.633 kN to 18.705 kN. From these results, it can be seen a square hole as best design in obtaining the smallest maximum impact force

Deep splicing plasticity of the human adenovirus type 5 transcriptome drives virus evolution

Viral genomes have high gene densities and complex transcription strategies rendering transcriptome analysis through short-read RNA-seq approaches problematic. Adenovirus transcription and splicing is especially complex. We used long-read direct RNA sequencing to study adenovirus transcription and splicing during infection. This revealed a previously unappreciated complexity of alternative splicing and potential for secondary initiating codon usage. Moreover, we find that most viral transcripts tend to shorten polyadenylation lengths as infection progresses. Development of an open reading frame centric bioinformatics analysis pipeline provided a deeper quantitative and qualitative understanding of adenovirus’s genetic potential. Across the viral genome adenovirus makes multiple distinctly spliced transcripts that code for the same protein. Over 11,000 different splicing patterns were recorded across the viral genome, most occurring at low levels. This low-level use of alternative splicing patterns potentially enables the virus to maximise its coding potential over evolutionary timescales

Direct RNA sequencing of respiratory syncytial virus infected human cells generates a detailed overview of RSV polycistronic mRNA and transcript abundance

To characterize species of viral mRNA transcripts generated during respiratory syncytial virus (RSV) infection, human fibroblast-like MRC-5 lung cells were infected with subgroup A RSV for 6, 16 and 24 hours. In addition, we characterised the viral transcriptome in infected Calu-3 lung epithelial cells at 48 hours post infection. Total RNA was harvested and polyadenylated mRNA was enriched and sequenced by direct RNA sequencing using an Oxford nanopore device. This platform yielded over 450,000 direct mRNA transcript reads which were mapped to the viral genome and analysed to determine the relative mRNA levels of viral genes using our in-house ORF-centric pipeline. We examined the frequency of polycistronic readthrough mRNAs were generated and assessed the length of the polyadenylated tails for each group of transcripts. We show a general but non-linear decline in gene transcript abundance across the viral genome, as predicted by the model of RSV gene transcription. However, the decline in transcript abundance is not uniform. The polyadenylate tails generated by the viral polymerase are similar in length to those generated by the host polyadenylation machinery and broadly declined in length for most transcripts as the infection progressed. Finally, we observed that the steady state abundance of transcripts with very short polyadenylate tails less than 20 nucleotides is less for N, SH and G transcripts in both cell lines compared to NS1, NS2, P, M, F and M2 which may reflect differences in mRNA stability and/or translation rates within and between the cell lines

Direct RNA sequencing of respiratory syncytial virus infected human cells generates a detailed overview of RSV polycistronic mRNA and transcript abundance.

To characterize species of viral mRNA transcripts generated during respiratory syncytial virus (RSV) infection, human fibroblast-like MRC-5 lung cells were infected with subgroup A RSV for 6, 16 and 24 hours. In addition, we characterised the viral transcriptome in infected Calu-3 lung epithelial cells at 48 hours post infection. Total RNA was harvested and polyadenylated mRNA was enriched and sequenced by direct RNA sequencing using an Oxford nanopore device. This platform yielded over 450,000 direct mRNA transcript reads which were mapped to the viral genome and analysed to determine the relative mRNA levels of viral genes using our in-house ORF-centric pipeline. We examined the frequency of polycistronic readthrough mRNAs were generated and assessed the length of the polyadenylated tails for each group of transcripts. We show a general but non-linear decline in gene transcript abundance across the viral genome, as predicted by the model of RSV gene transcription. However, the decline in transcript abundance is not uniform. The polyadenylate tails generated by the viral polymerase are similar in length to those generated by the host polyadenylation machinery and broadly declined in length for most transcripts as the infection progressed. Finally, we observed that the steady state abundance of transcripts with very short polyadenylate tails less than 20 nucleotides is less for N, SH and G transcripts in both cell lines compared to NS1, NS2, P, M, F and M2 which may reflect differences in mRNA stability and/or translation rates within and between the cell lines

The P323L substitution in the SARS-CoV-2 polymerase (NSP12) confers a selective advantage during infection

Background: The mutational landscape of SARS-CoV-2 varies at the dominant viral genome sequence and minor genomic variant population. During the COVID-19 pandemic, an early substitution in the genome was the D614G change in the spike protein, associated with an increase in transmissibility. Genomes with D614G are accompanied by a P323L substitution in the viral polymerase (NSP12). However, P323L is not thought to be under strong selective pressure. Results: Investigation of P323L/D614G substitutions in the population shows rapid emergence during the containment phase and early surge phase during the first wave. These substitutions emerge from minor genomic variants which become dominant viral genome sequence. This is investigated in vivo and in vitro using SARS-CoV-2 with P323 and D614 in the dominant genome sequence and L323 and G614 in the minor variant population. During infection, there is rapid selection of L323 into the dominant viral genome sequence but not G614. Reverse genetics is used to create two viruses (either P323 or L323) with the same genetic background. L323 shows greater abundance of viral RNA and proteins and a smaller plaque morphology than P323. Conclusions: These data suggest that P323L is an important contribution in the emergence of variants with transmission advantages. Sequence analysis of viral populations suggests it may be possible to predict the emergence of a new variant based on tracking the frequency of minor variant genomes. The ability to predict an emerging variant of SARS-CoV-2 in the global landscape may aid in the evaluation of medical countermeasures and non-pharmaceutical interventions

Molnupiravir versus placebo in unvaccinated and vaccinated patients with early SARS-CoV-2 infection in the UK (AGILE CST-2): a randomised, placebo-controlled, double-blind, phase 2 trial.

The antiviral drug molnupiravir was licensed for treating at-risk patients with COVID-19 on the basis of data from unvaccinated adults. We aimed to evaluate the safety and virological efficacy of molnupiravir in vaccinated and unvaccinated individuals with COVID-19. This randomised, placebo-controlled, double-blind, phase 2 trial (AGILE CST-2) was done at five National Institute for Health and Care Research sites in the UK. Eligible participants were adult (aged ≥18 years) outpatients with PCR-confirmed, mild-to-moderate SARS-CoV-2 infection who were within 5 days of symptom onset. Using permuted blocks (block size 2 or 4) and stratifying by site, participants were randomly assigned (1:1) to receive either molnupiravir (orally; 800 mg twice daily for 5 days) plus standard of care or matching placebo plus standard of care. The primary outcome was the time from randomisation to SARS-CoV-2 PCR negativity on nasopharyngeal swabs and was analysed by use of a Bayesian Cox proportional hazards model for estimating the probability of a superior virological response (hazard ratio [HR]&gt;1) for molnupiravir versus placebo. Our primary model used a two-point prior based on equal prior probabilities (50%) that the HR was 1·0 or 1·5. We defined a priori that if the probability of a HR of more than 1 was more than 80% molnupiravir would be recommended for further testing. The primary outcome was analysed in the intention-to-treat population and safety was analysed in the safety population, comprising participants who had received at least one dose of allocated treatment. This trial is registered in ClinicalTrials.gov, NCT04746183, and the ISRCTN registry, ISRCTN27106947, and is ongoing. Between Nov 18, 2020, and March 16, 2022, 1723 patients were assessed for eligibility, of whom 180 were randomly assigned to receive either molnupiravir (n=90) or placebo (n=90) and were included in the intention-to-treat analysis. 103 (57%) of 180 participants were female and 77 (43%) were male and 90 (50%) participants had received at least one dose of a COVID-19 vaccine. SARS-CoV-2 infections with the delta (B.1.617.2; 72 [40%] of 180), alpha (B.1.1.7; 37 [21%]), omicron (B.1.1.529; 38 [21%]), and EU1 (B.1.177; 28 [16%]) variants were represented. All 180 participants received at least one dose of treatment and four participants discontinued the study (one in the molnupiravir group and three in the placebo group). Participants in the molnupiravir group had a faster median time from randomisation to negative PCR (8 days [95% CI 8-9]) than participants in the placebo group (11 days [10-11]; HR 1·30, 95% credible interval 0·92-1·71; log-rank p=0·074). The probability of molnupiravir being superior to placebo (HR&gt;1) was 75·4%, which was less than our threshold of 80%. 73 (81%) of 90 participants in the molnupiravir group and 68 (76%) of 90 participants in the placebo group had at least one adverse event by day 29. One participant in the molnupiravir group and three participants in the placebo group had an adverse event of a Common Terminology Criteria for Adverse Events grade 3 or higher severity. No participants died (due to any cause) during the trial. We found molnupiravir to be well tolerated and, although our predefined threshold was not reached, we observed some evidence that molnupiravir has antiviral activity in vaccinated and unvaccinated individuals infected with a broad range of SARS-CoV-2 variants, although this evidence is not conclusive.</p

Genome evolution of dengue virus serotype 1 under selection by <i>Wolbachia pipientis</i> in <i>Aedes aegypti</i> mosquitoes.

The introgression of antiviral strains of Wolbachia into Aedes aegypti mosquito populations is a public health intervention for the control of dengue. Plausibly, dengue virus (DENV) could evolve to bypass the antiviral effects of Wolbachia and undermine this approach. Here, we established a serial-passage system to investigate the evolution of DENV in Ae. aegypti mosquitoes infected with the wMel strain of Wolbachia. Using this system, we report on virus genetic outcomes after twenty passages of serotype 1 of DENV (DENV-1). An amino acid substitution, E203K, in the DENV-1 envelope protein was more frequently detected in the consensus sequence of virus populations passaged in wMel-infected Ae. aegypti than wild-type counterparts. Positive selection at residue 203 was reproducible; it occurred in passaged virus populations from independent DENV-1-infected patients and also in a second, independent experimental system. In wild-type mosquitoes and human cells, the 203K variant was rapidly replaced by the progenitor sequence. These findings provide proof of concept that wMel-associated selection of virus populations can occur in experimental conditions. Field-based studies are needed to explore whether wMel imparts selective pressure on DENV evolution in locations where wMel is established

Characterising the genomic stability of nucleoside analogue-treated SARS-CoV-2.

Therapeutics that have broad-spectrum antiviral activity are crucial in a pandemic preparedness toolkit. When the SARS-CoV-2 pandemic began, many antiviral drugs were repurposed, and new ones were discovered. However, despite promising results from in vitro and animal models, many of the tested repurposed drugs were clinically ineffective. Detailed characterisation was necessary to assess how nucleoside analogues, a class of broad-spectrum antivirals, induce lethal mutagenesis. This focused on the impact on the genomic stability of SARS-CoV-2 and the potential longevity of the clinical administration of these drugs. This thesis evaluates two nucleoside analogues, molnupiravir and favipiravir, and the likelihood of SARS-CoV-2 developing resistance to them when widely used. This thesis confirmed the mechanism of action of molnupiravir in human patients, which was lacking in the scientific literature at the time. High-throughput amplicon sequencing, minor genomic variant analysis, and bespoke data visualisation pipelines were used to analyse longitudinal nasopharyngeal samples from the AGILE coronavirus drug testing initiative's phase II molnupiravir efficacy trial. The study found no indication of antiviral resistance markers in the time points analysed. The data from this trial was used as a control dataset in a global investigation into clusters of SARS-CoV-2 sequences with strong molnupiravir-like mutation signals appearing in sequence databases, some of which showed evidence of seeding onward transmission. Overall, the data from this thesis established a clear rubric for assessing the evolutionary safety of molnupiravir, which can inform public health, genomic surveillance, and molnupiravir licensing and regulatory approvals worldwide. Experimental evolution studies were conducted to elucidate why SARS-CoV-2 viability might persist and lead to transmission even after drug treatment. The study demonstrated the molnupiravir-induced mutagenic tolerability of SARS-CoV-2 ancestral and Delta lineages, particularly in the spike gene. Although less extensively authorised than molnupiravir, favipiravir has been widely used to treat COVID-19 patients in several countries, including Japan and Saudi Arabia. Consequently, a similar in vitro study was conducted to examine the effect of favipiravir on the genomic stability of XBB.1.1, an Omicron sub-lineage. This study identified the emergence of two RNA-dependent RNA polymerase mutations in nsp12, which have potential implications for antiviral resistance. The research described in this thesis has contributed critical data on how nucleoside analogues affect the evolution of SARS-CoV-2. This information can be used to develop bioinformatic tools tailored to different antiviral drug modalities to predict and monitor the incidence of antiviral drug resistance and other evolutionary consequences