The surface accessibility of the glycine receptor M2-M3 loop is increased in the channel open state

Mutations in the extracellular M2-M3 loop of the glycine receptor (GlyR) alpha1 subunit have been shown previously to affect channel gating. In this study, the substituted cysteine accessibility method was used to investigate whether a structural rearrangement of the M2-M3 loop accompanies GlyR activation. All residues from R271C to V277C were covalently modified by both positively charged methanethiosulfonate ethyltrimethylammonium (MTSET) and negatively charged methanethiosulfonate ethylsulfonate (MTSES), implying that these residues form an irregular surface loop. The MTSET modification rate of all residues from R271C to K276C was faster in the glycine-bound state than in the unliganded state. MTSES modification of A272C, L274C, and V277C was also faster in the glycine-bound state. These results demonstrate that the surface accessibility of the M2-M3 loop is increased as the channel transitions from the closed to the open state, implying that either the loop itself or an overlying domain moves during channel activation

Both Size and GC-Content of Minimal Introns Are Selected in Human Populations

Background: We previously have studied the insertion and deletion polymorphism by sequencing no more than one hundred introns in a mixed human population and found that the minimal introns tended to maintain length at an optimal size. Here we analyzed re-sequenced 179 individual genomes (from African, European, and Asian populations) from the data released by the 1000 Genome Project to study the size dynamics of minimal introns. Principal Findings: We not only confirmed that minimal introns in human populations are selected but also found two major effects in minimal intron evolution: (i) Size-effect: minimal introns longer than an optimal size (87 nt) tend to have a higher ratio of deletion to insertion than those that are shorter than the optimal size; (ii) GC-effect: minimal introns with lower GC content tend to be more frequently deleted than those with higher GC content. The GC-effect results in a higher GC content in minimal introns than their flanking exons as opposed to larger introns ($125 nt) that always have a lower GC content than that of their flanking exons. We also observed that the two effects are distinguishable but not completely separable within and between populations. Conclusions: We validated the unique mutation dynamics of minimal introns in keeping their near-optimal size and GC content, and our observations suggest potentially important functions of human minimal introns in transcript processin

Effectiveness of a physical activity pilot intervention in youth with type 1 diabetes : the ActivPals study

Background: Type 1 Diabetes (T1D) is rising globally. Youth with T1D continue to suffer from poorer health and lower levels of physical activity (PA) than those without diabetes. The ActivPals study aimed to support young people with T1D to increase PA levels. Methods: 20 youth (aged 7-16) with T1D were randomised to a pilot PA intervention (n=10) or control group. The Actigraph GT3X+ monitor measured PA at baseline and one-month follow-up to test the effectiveness of the intervention. PedsQol scales (generic and diabetes module) were used to measure Quality of Life (QoL) in participants and parents at baseline and follow- up. Results: Changes in PA in QoL were analysed using a two- way mixed ANOVA. The results showed a significant increase in Moderate to Vigorous PA (MVPA) in both intervention and control group from baseline to follow up (p= 0.03), however there were no significant between group differences. Both groups reported significantly less overall diabetes ‘problems’ (p=0.012) and significantly less lifestyle ‘problems’ (p= 0.015) at follow up. However, intervention and control participants also reported significant increases over time in ‘problems’ with daily diabetes routine (p= 0.022). Parents reported increased worry about their child’s diabetes at follow up, significant across both groups (p= 0.046). There was no significant increase in reported hypoglycaemic occurrences despite increased MVPA. Conclusions: A larger scale trial, with longer intervention period could significantly increase the MVPA levels and QoL in youth with T1D without significantly increasing hypoglycaemic episodes

High genetic diversity at the extreme range edge: nucleotide variation at nuclear loci in Scots pine (Pinus sylvestris L.) in Scotland

Nucleotide polymorphism at 12 nuclear loci was studied in Scots pine populations across an environmental gradient in Scotland, to evaluate the impacts of demographic history and selection on genetic diversity. At eight loci, diversity patterns were compared between Scottish and continental European populations. At these loci, a similar level of diversity (θsil=~0.01) was found in Scottish vs mainland European populations, contrary to expectations for recent colonization, however, less rapid decay of linkage disequilibrium was observed in the former (ρ=0.0086±0.0009, ρ=0.0245±0.0022, respectively). Scottish populations also showed a deficit of rare nucleotide variants (multi-locus Tajima's D=0.316 vs D=−0.379) and differed significantly from mainland populations in allelic frequency and/or haplotype structure at several loci. Within Scotland, western populations showed slightly reduced nucleotide diversity (πtot=0.0068) compared with those from the south and east (0.0079 and 0.0083, respectively) and about three times higher recombination to diversity ratio (ρ/θ=0.71 vs 0.15 and 0.18, respectively). By comparison with results from coalescent simulations, the observed allelic frequency spectrum in the western populations was compatible with a relatively recent bottleneck (0.00175 × 4Ne generations) that reduced the population to about 2% of the present size. However, heterogeneity in the allelic frequency distribution among geographical regions in Scotland suggests that subsequent admixture of populations with different demographic histories may also have played a role

Lockdown labs : pivoting to remote learning in forensic science higher education

Forensic Science training and education is reliant on the application of knowledge to casework scenarios and the development of key practical skills that provide a platform for career development in the field. The COVID-19 pandemic introduced a number of challenges to effectively deliver practical content online and remotely, whilst still meeting intended learning outcomes, accreditation requirements, and attaining a high level of student engagement and experience. The MSc Forensic Science programme featured in this study is a one-year degree programme with a strong emphasis on the practical elements of forensic science, and a diverse international student cohort. Therefore, the restrictions associated with the pandemic made it very difficult not only to plan the delivery of material but also to adapt the content itself for effective online and remote learning. By focusing on the intended learning outcomes, a number of innovative teaching practices were developed to successfully transition from face-to-face teaching to online and remote delivery. A range of online and practical resources were developed, including a laboratory home kit, demonstration videos, online practical technique simulations (produced by Learning Science), data analysis tasks, and interactive workshops and activities, all designed to consolidate student learning and build confidence, in preparation for such a time that on campus practical teaching could resume. The initial feedback received from these activities from both staff and students was extremely positive and the transition from classroom to online teaching was a success, as reflected in student attainment and later student feedback. Students reported that they had a better understanding of what was expected of them, including knowledge of protocols and techniques, and felt much more confident moving into the next stage of their learning development. Even though the practical laboratory sessions were the most significantly affected by the restrictions associated with the pandemic, and resulted in reduced interaction for the students, this was counteracted by virtual sessions and workshops, which gave students the opportunity to engage with each other and communicate their thoughts and opinions, ultimately building key presentation and group working skills. This case study will detail the pivot to remote learning, as well as critically evaluating the feedback from students and discussing the changes that are likely to be retained as longer-term teaching practices, versus those that were a necessary temporary addition or adjustment in response to the pandemic

Complete chloroplast genome sequence of Holoparasite Cistanche Deserticola (Orobanchaceae) reveals gene loss and horizontal gene transfer from Its host Haloxylon Ammodendron (Chenopodiaceae)

The central function of chloroplasts is to carry out photosynthesis, and its gene content and structure are highly conserved across land plants. Parasitic plants, which have reduced photosynthetic ability, suffer gene losses from the chloroplast (cp) genome accompanied by the relaxation of selective constraints. Compared with the rapid rise in the number of cp genome sequences of photosynthetic organisms, there are limited data sets from parasitic plants. The authors report the complete sequence of the cp genome of Cistanche deserticola, a holoparasitic desert species belonging to the family Orobanchaceae

Research trends in forensic science:A scientometric approach to analyze the content of the INTERPOL reviews

The use of forensic evidence has become indispensable in many countries and jurisdictions around the world, however the dissemination of research advancements does not necessarily directly or easily reach the forensic science community. Reports from the INTERPOL International Forensic Science Managers Symposium outline major areas that are of interest to forensic practitioners across the INTERPOL member countries. The information contained in the INTERPOL reports is extensive but can be challenging to process. The purpose of this research is to provide a comprehensive overview of the evolution of trends within the INTERPOL reports over an 18 year period. References relating to 10 evidence types retrieved from the 14th to 19th INTERPOL IFSMS reports (2004-2019) were processed and compared with data exports from the citation database Scopus covering the same evidence types. The results from this work are summarised by investigating the relationships between the 10 evidence types. To explore the outputs a user-friendly R-Shiny application was developed and is freely available at: https://uod.ac.uk/lrcfsinterpolreportsexplorer

Large introns in relation to alternative splicing and gene evolution: a case study of Drosophila bruno-3

Background: Alternative splicing (AS) of maturing mRNA can generate structurally and functionally distinct transcripts from the same gene. Recent bioinformatic analyses of available genome databases inferred a positive correlation between intron length and AS. To study the interplay between intron length and AS empirically and in more detail, we analyzed the diversity of alternatively spliced transcripts (ASTs) in the Drosophila RNA-binding Bruno-3 (Bru-3) gene. This gene was known to encode thirteen exons separated by introns of diverse sizes, ranging from 71 to 41,973 nucleotides in D. melanogaster. Although Bru-3's structure is expected to be conducive to AS, only two ASTs of this gene were previously described. Results: Cloning of RT-PCR products of the entire ORF from four species representing three diverged Drosophila lineages provided an evolutionary perspective, high sensitivity, and long-range contiguity of splice choices currently unattainable by high-throughput methods. Consequently, we identified three new exons, a new exon fragment and thirty-three previously unknown ASTs of Bru-3. All exon-skipping events in the gene were mapped to the exons surrounded by introns of at least 800 nucleotides, whereas exons split by introns of less than 250 nucleotides were always spliced contiguously in mRNA. Cases of exon loss and creation during Bru-3 evolution in Drosophila were also localized within large introns. Notably, we identified a true de novo exon gain: exon 8 was created along the lineage of the obscura group from intronic sequence between cryptic splice sites conserved among all Drosophila species surveyed. Exon 8 was included in mature mRNA by the species representing all the major branches of the obscura group. To our knowledge, the origin of exon 8 is the first documented case of exonization of intronic sequence outside vertebrates. Conclusion: We found that large introns can promote AS via exon-skipping and exon turnover during evolution likely due to frequent errors in their removal from maturing mRNA. Large introns could be a reservoir of genetic diversity, because they have a greater number of mutable sites than short introns. Taken together, gene structure can constrain and/or promote gene evolution

Recent and Recurrent Selective Sweeps of the Antiviral RNAi Gene Argonaute-2 in Three Species of Drosophila

Antagonistic host–parasite interactions can drive rapid adaptive evolution in genes of the immune system, and such arms races may be an important force shaping polymorphism in the genome. The RNA interference pathway gene Argonaute-2 (AGO2) is a key component of antiviral defense in Drosophila, and we have previously shown that genes in this pathway experience unusually high rates of adaptive substitution. Here we study patterns of genetic variation in a 100-kbp region around AGO2 in three different species of Drosophila. Our data suggest that recent independent selective sweeps in AGO2 have reduced genetic variation across a region of more than 50 kbp in Drosophila melanogaster, D. simulans, and D. yakuba, and we estimate that selection has fixed adaptive substitutions in this gene every 30–100 thousand years. The strongest signal of recent selection is evident in D. simulans, where we estimate that the most recent selective sweep involved an allele with a selective advantage of the order of 0.5–1% and occurred roughly 13–60 Kya. To evaluate the potential consequences of the recent substitutions on the structure and function of AGO2, we used fold-recognition and homology-based modeling to derive a structural model for the Drosophila protein, and this suggests that recent substitutions in D. simulans are overrepresented at the protein surface. In summary, our results show that selection by parasites can consistently target the same genes in multiple species, resulting in areas of the genome that have markedly reduced genetic diversity

Population Genomic Inferences from Sparse High-Throughput Sequencing of Two Populations of Drosophila melanogaster

Short-read sequencing techniques provide the opportunity to capture genome-wide sequence data in a single experiment. A current challenge is to identify questions that shallow-depth genomic data can address successfully and to develop corresponding analytical methods that are statistically sound. Here, we apply the Roche/454 platform to survey natural variation in strains of Drosophila melanogaster from an African (n = 3) and a North American (n = 6) population. Reads were aligned to the reference D. melanogaster genomic assembly, single nucleotide polymorphisms were identified, and nucleotide variation was quantified genome wide. Simulations and empirical results suggest that nucleotide diversity can be accurately estimated from sparse data with as little as 0.2× coverage per line. The unbiased genomic sampling provided by random short-read sequencing also allows insight into distributions of transposable elements and copy number polymorphisms found within populations and demonstrates that short-read sequencing methods provide an efficient means to quantify variation in genome organization and content. Continued development of methods for statistical inference of shallow-depth genome-wide sequencing data will allow such sparse, partial data sets to become the norm in the emerging field of population genomics