Excess Dally-like Induces Malformation of Drosophila Legs.

Glypicans are closely associated with organ development and tumorigenesis in animals. Dally-like (Dlp), a membrane-bound glypican, plays pivotal roles in various biological processes in Drosophila. In this study, we observed that an excess of Dlp led to the malformation of legs, particularly affecting the distal part. Accordingly, the leg disc was shrunken and frequently exhibited aberrant morphology. In addition, elevated Dlp levels induced ectopic cell death with no apparent cell proliferation changes. Furthermore, Dlp overexpression in the posterior compartment significantly altered Wingless (Wg) distribution. We observed a marked expansion of Wg distribution within the posterior compartment, accompanied by a corresponding decrease in the anterior compartment. It appears that excess Dlp guides Wg to diffuse to cells with higher Dlp levels. In addition, the distal-less (dll) gene, which is crucial for leg patterning, was up-regulated significantly. Notably, dachshund (dac) and homothorax (hth) expression, also essential for leg patterning and development, only appeared to be negligibly affected. Based on these findings, we speculate that excess Dlp may contribute to malformations of the distal leg region of Drosophila, possibly through its influence on Wg distribution, dll expression and induced cell death. Our research advances the understanding of Dlp function in Drosophila leg development.

Wnt/Wingless signaling promotes lipid mobilization through signal-induced transcriptional repression.

The Wnt/Wingless signaling pathway plays critical roles in metazoan development and energy metabolism, but its role in regulating lipid homeostasis remains not fully understood. Here, we report that the activation of canonical Wnt/Wg signaling promotes lipolysis while concurrently inhibiting lipogenesis and fatty acid ß-oxidation in both larval and adult adipocytes, as well as cultured S2R+ cells, in Drosophila. Using RNA-sequencing and CUT&RUN (Cleavage Under Targets & Release Using Nuclease) assays, we identified a set of Wnt target genes responsible for intracellular lipid homeostasis. Notably, active Wnt signaling directly represses the transcription of these genes, resulting in decreased de novo lipogenesis and fatty acid ß-oxidation, but increased lipolysis. These changes lead to elevated free fatty acids and reduced triglyceride (TG) accumulation in adipocytes with active Wnt signaling. Conversely, downregulation of Wnt signaling in the fat body promotes TG accumulation in both larval and adult adipocytes. The attenuation of Wnt signaling also increases the expression of specific lipid metabolism-related genes in larval adipocytes, wing discs, and adult intestines. Taken together, these findings suggest that Wnt signaling-induced transcriptional repression plays an important role in regulating lipid homeostasis by enhancing lipolysis while simultaneously suppressing lipogenesis and fatty acid ß-oxidation.

A dataset of CMIP6-based climate scenarios for climate change impact assessment in Great Britain.

Climate change is a critical issue in the 21st century. Assessment of the impacts of climate change is beneficial for assisting advanced recommendations for adaptations. Climate change impact assessments require high quality local-scale climate scenarios. The future climate projections from Global Climate Models (GCMs) are problematic to use at local scale due to their coarse spatial and temporal resolution, and existing biases. It is important to have climate change scenarios based on the GCMs ensemble downscaled to local scale to account for inherent uncertainty in climate projections, and to have a sufficient large number of years to account for inter-annual climate variability and low frequency, but high impact, extreme climatic events. A dataset of future climate change scenarios was therefore generated at 26 representative sites across Great Britain based on the latest CMIP6 multi-model ensemble downscaled to local-scale by using a stochastic weather generator, LARS-WG 8.0. The data set consists of climate scenarios of daily weather of 1,000 realizations of typical years for a baseline, and very near (2030) and near-future (2050) climates, based on five GCMs and two emission scenarios (Shared Socioeconomic Pathways - SSPs viz. SSP2-4.5 and SSP5-8.5). A total of 15 GCMs from the CMIP6 ensemble were integrated in LARS-WG 8.0. LARS-WG downscales future climate projections from the GCMs and incorporates changes at local scale in the mean climate, climatic variability, and extreme events by modifying the statistical distributions of the weather variables at each site. Based on the performance of the GCMs over northern Europe and their climate sensitivity, a subset of five GCMs was selected, viz.; ACCESS-ESM1-5, CNRM-CM6-1, HadGEM3-GC31-LL, MPI-ESM1-2-LR and MRI-ESM2-0. The selected GCMs are evenly distributed among the full set of 15 GCMs. The use of a subset of GCMs substantially reduces computational time, while allowing assessment of uncertainties in impact studies related to uncertain future climate projections arising from GCMs. The 1000 years of daily weather for the baseline, as well as for very near and near-future climate change scenarios, are essential for estimating inter-annual variation, and for detecting low frequency, but high impact, extreme climatic events, such as heat waves, floods and droughts. The present dataset can be used as an input to climate change impact models in various fields, including, land and water resources, agriculture and food production, ecology and epidemiology, and human health and welfare. Researchers, breeders, farm managers, social and public sector leaders, and policymakers may benefit from this new dataset when undertaking impact assessments of climate change and decision support for mitigation and adaptation to climate change.

Pygo-F773W Mutation Reveals Novel Functions beyond Wnt Signaling in Drosophila.

Pygopus (Pygo) has been identified as a specific nuclear co-activator of the canonical Wingless (Wg)/Wnt signaling pathway in Drosophila melanogaster. Pygo proteins consist of two conserved domains: an N-terminal homologous domain (NHD) and a C-terminal plant homologous domain (PHD). The PHD's ability to bind to di- and trimethylated lysine 4 of histone H3 (H3K4me2/3) appears to be independent of Wnt signaling. There is ongoing debate regarding the significance of Pygo's histone-binding capacity. Drosophila Pygo orthologs have a tryptophan (W) > phenylalanine (F) substitution in their histone pocket-divider compared to vertebrates, leading to reduced histone affinity. In this research, we utilized CRISPR/Cas9 technology to introduce the Pygo-F773W point mutation in Drosophila, successfully establishing a viable homozygous Pygo mutant line for the first time. Adult mutant flies displayed noticeable abnormalities in reproduction, locomotion, heart function, and lifespan. RNA-seq and cluster analysis indicated that the mutation primarily affected pathways related to immunity, metabolism, and posttranslational modification in adult flies rather than the Wnt signaling pathway. Additionally, a reduction in H3K9 acetylation levels during the embryonic stage was observed in the mutant strains. These findings support the notion that Pygo plays a wider role in chromatin remodeling, with its involvement in Wnt signaling representing only a specific aspect of its chromatin-related functions.

Broad synergistic antiviral efficacy between a novel elite controller-derived dipeptide and antiretrovirals against drug-resistant HIV-1.

Introduction: The naturally occurring dipeptide Tryptophylglycine (WG) is enhanced in human immunodeficiency virus (HIV-1) infected Elite Controllers (EC). We have shown that this dipeptide has an anti-HIV-1 effect and evaluated now its synergistic antiretroviral activity, in combination with current antiretrovirals against multi-drug resistant HIV-1 isolates. Methods: Drug selectivity assay with WG-am and ARVs agains HIV-1 resistant isolates were carried out. Subsequently, two methods, Chou-Talalay's Combination Index (CI) and ZIP synergy score (SS), were used to quantify the synergism. Results: WG-am had a moderate/strong synergism with the four tested antiretrovirals: raltegravir, tenofovir, efavirenz, darunavir. WG-am:TDF had strong synergism at ED50, ED75, ED90 (CI: <0.2) in isolates resistant to protease inhibitors or integrase strand inhibitors (INSTI), and a slightly less synergism in isolates resistant to non-nucleoside or nucleotide reverse transcriptase inhibitors. WG-am combined with each of the four drugs inhibited all drug-resistant isolates with over 95% reduction at maximum concentration tested. The highest selectivity indexes (CC50/ED50) were in INSTI-resistant isolates. Conclusion: Our data suggest that WG, identified as occurring and enhanced in Elite Controllers has a potential to become a future treatment option in patients with HIV-1 strains resistant to any of the four major categories of anti-HIV-1 compounds.

Ecotoxicological Evaluation of Safener and Antimicrobial Additives in Isoxaflutole-Based Herbicide Formulations.

The environmental load by isoxaflutole and its formulated herbicide products has increasingly become apparent because, after the ban of atrazine, isoxaflutole has become its replacement active ingredient (a.i.). Obtaining information regarding the fate of this a.i. in environmental matrices and its ecotoxicological effects on aquatic organisms is essential for the risk assessment of the herbicide. In this study, the effects of Merlin Flexx- and Merlin WG75 formulated isoxaflutole-based herbicide products and two selected additives (cyprosulfamide safener and 1,2-benzisothiazol-3(2H)-one antimicrobial agent) were investigated on Raphidocelis subcapitata in growth inhibition assays. In ecotoxicological tests, two conventional (optical density and chlorophyll-a content) and two induced fluorescence-based (Fv*/Fp: efficiency of the photosystem PSII and Rfd* changes in the observed ratio of fluorescence decrease) endpoints were determined by UV-spectrophotometer and by our FluoroMeter Module, respectively. Furthermore, dissipation of isoxaflutole alone and in its formulated products was examined by an HPLC-UV method. In ecotoxicological assays, the fluorescence-based Rfd* was observed as the most sensitive endpoint. In this study, the effects of the safener cyprosulfamide and the antimicrobial agent 1,2-benzisothiazol-3(2H)-one on R. subcapitata is firstly reported. The results indicated that the isoxaflutole-equivalent toxicity of the mixture of the isoxaflutole-safener-antimicrobial agent triggered lower toxicity (EC50 = 2.81 ± 0.22 mg/L) compared to the individual effect of the a.i. (EC50 = 0.02 ± 0.00 mg/L). The Merlin Flexx formulation (EC50 = 27.04 ± 1.41 mg/L) was found to be approximately 50-fold less toxic than Merlin WG75, which can be explained by the different chemical characteristics and quantity of additives in them. The additives influenced the dissipation of the a.i. in Z8 medium, as the DT50 value decreased by approximately 1.2- and 3.5-fold under light and dark conditions, respectively.

Determination of the larval precursor configuration of the Drosophila adult hindgut by G-TRACE analysis.

The Drosophila hindgut is a classical model to study organogenesis. The adult hindgut originates from the precursor cells in the larval hindgut. However, the territory of these cells has still not been well determined. A ring of wingless (wg)-expressing cells lies at the anterior zone of both the larval and adult hindgut. The larval Wg ring was thought as a portion of precursor of the adult hindgut. By applying a cell lineage tracing tool (G-TRACE), we demonstrate that larval wg-expressing cells have no cell lineage contribution to the adult hindgut. Additionally, adult Wg ring cells do not divide and move posteriorly to replenish the hindgut tissue. Instead, we determine that the precursors of the adult pylorus and ileum are situated in the cubitus interruptus (ci)-expressing cells in the anterior zone, and deduce that the precursor stem cells of the adult rectum locate in the trunk region of the larval pylorus including hedgehog (hh)-expressing cells. Together, this research advances our understanding of cell lineage origins and the development of the Drosophila hindgut.

Quantitative analysis of the protective performance of bicycle helmet with multi-direction impact protection system in oblique impact tests.

PURPOSE: The current study aimed to assess the protective performance of helmets equipped with multi-directional impact protection system (MIPS) under various oblique impact loads. METHODS: Initially, a finite element model of a bicycle helmet with MIPS was developed based on the scanned geometric parameters of an actual bicycle helmet. Subsequently, the validity of model was confirmed using the KASK WG11 oblique impact test method. Three different impact angles (30°, 45°, and 60°) and 2 varying impact speeds (5 m/s and 8 m/s) were employed in oblique tests to evaluate protective performance of MIPS in helmets, focusing on injury assessment parameters such as peak linear acceleration (PLA) and peak angular acceleration (PAA) of the head. RESULTS: The results demonstrated that in all impact simulations, both assessment parameters were lower during impact for helmets equipped with MIPS compared to those without. The PAA was consistently lower in the MIPS helmet group, whereas the difference in PLA was not significant in the no-MIPS helmet group. For instance, at an impact velocity of 8 m/s and a 30° inclined anvil, the MIPS helmet group exhibited a PAA of 3225 rad/s2 and a PLA of 281 g. In contrast, the no-MIPS helmet group displayed a PAA of 8243 rad/s2 and a PLA of 292 g. Generally, both PAA and PLA parameters decreased with the increase of anvil angles. At a 60° anvil angles, PAA and PLA values were 664 rad/s2 and 20.7 g, respectively, reaching their minimum. CONCLUSION: The findings indicated that helmets incorporating MIPS offer enhanced protection against various oblique impact loads. When assessing helmets for oblique impacts, the utilization of larger angle anvils and rear impacts might not adequately evaluate protective performance during an impact event. These findings will guide advancements in helmet design and the refinement of oblique impact test protocols.

Cognitive Testing and Piloting of the Bangla Version of the Washington Group Short Set Questionnaire on Functioning Among Adolescent Girls and Women With Disabilities in Selected Sub-districts of Bangladesh.

Introduction This study focuses on the cognitive testing and piloting of the Bangla version of the Washington Group Short Set Questionnaire on Functioning among adolescent girls and women with disabilities in selected sub-districts of Bangladesh. The Washington Group on Disability Statistics developed the questionnaire as a tool to assess the functioning and disability status of individuals. The adaptation of this questionnaire to Bangla is crucial for capturing accurate data on the experiences of adolescent girls and women with disabilities in Bangladesh. Materials and methods The research employs a two-phase approach, starting with cognitive testing to ensure the linguistic and cultural relevance of the translated questionnaire. This phase involves engaging with a sample of the target population to assess the comprehension, clarity, and appropriateness of the questions. Subsequently, a pilot study was conducted in selected sub-districts to evaluate the feasibility and validity of the Bangla version of the Washington Group Short Set Questionnaire on Functioning in real-world settings. Both of the tests were conducted in March 2023. Results There were different types of participants with different types of disabilities. Information processing, meaning, understanding the questions, thinking, and answering speed or time were different between groups, even though they were different from person to person. The initial assessments indicate strong consistency in responses. Participants demonstrated a favorable response rate, indicating potential effectiveness for broader implementation. Conclusion The current study aims to contribute to disability data collection methodologies, particularly in the context of adolescent girls and women in Bangladesh. The research seeks to empower policymakers, researchers, and advocacy groups with a robust instrument of disability screening. Researchers and clinicians may rely on our accurate and validated Washington Group Short Set Questionnaire on Functioning translation into Bangla when working with adolescent girls and women with disabilities.

Whole genome sequence-based molecular characterization of blood isolates of carbapenem-resistant Enterobacter cloacae complex from ICU patients in Kolkata, India, during 2017-2022: emergence of phylogenetically heterogeneous Enterobacter hormaechei subsp. xiangfangensis.

Blood-borne infections caused by the carbapenem-resistant Enterobacter cloacae complex (CR-ECC) are major public threats with respect to the challenges encountered during treatment. This study describes the whole genome sequencing-based molecular characteristics of blood isolates (n = 70) of CR-ECC from patients admitted to the intensive care unit of tertiary care hospitals in Kolkata, India, during 2017-2022 with respect to species identification, antimicrobial resistance (AMR) profiling, mechanism of drug resistance, and molecular subtypes. Vitek2 MALDI and species-specific PCR identified Enterobacter hormaechei subsp. xiangfangensis (47.14%) as the emerging CR-ECC subspecies in Kolkata. The predominating carbapenemase and extended-spectrum ß-lactamase genes found were blaNDM-1 (51.42%) and blaCTX-M-15 (27%), respectively. Besides, blaNDM-4, blaNDM-5, blaNDM-7, blaCMH-3, blaSFO-1, blaOXA-181, blaOXA-232, blaKPC-3, and blaDHA-7 genes were also detected, which were not previously reported from India. A multitude of Class 1 integrons (including In180, In4874, In4887, and In4888, which were novel) and plasmid replicon types (IncFIB, IncFII, IncX3, IncHI1-HI2, IncC, and IncR) involved in AMR dissemination were identified. Reverse transcription-PCR and western blot revealed that carbapenem resistance in non-carbapenemase-producing CR-ECC isolates was contributed by elevated levels of ampC, overexpression of acrAB, and loss of ompF. A total of 30 distinct sequence types (STs) were ascertained by multi-locus sequence typing; of which, ST2011, ST2018, ST2055, ST2721, and ST2722 were novel STs. Pulsed-field gel electrophoresis analysis showed heterogeneity (69 pulsotypes with a similarity coefficient of 48.40%) among the circulating isolates, suggesting multiple reservoirs of infections in humans. Phylogenetically and genetically diverse CR-ECC with multiple AMR mechanisms mandates close monitoring of nosocomial infections caused by these isolates to forestall the transmission and dissemination of AMR.IMPORTANCEThe emergence and extensive dissemination of the carbapenem-resistant Enterobacter cloacae complex (CR-ECC) have positioned it as a critical nosocomial global pathogen. The dearth of a comprehensive molecular study pertaining to CR-ECC necessitated this study, which is the first of its kind from India. Characterization of blood isolates of CR-ECC over the last 6 years revealed Enterobacter hormaechei subsp. xiangfangensis as the most prevalent subsp., exhibiting resistance to almost all antibiotics currently in use and harboring diverse transmissible carbapenemase genes. Besides the predominating blaNDM-1 and blaCTX-M-15, we document diverse carbapenemase and AmpC genes, such as blaNDM-4, blaNDM-7, blaOXA-181, blaOXA-232, blaKPC-3, blaCMH-3, blaSFO-1, and blaDHA-7, in CR-ECC, which were not previously reported from India. Furthermore, novel integrons and sequence types were identified. Our findings emphasize the need for strengthened vigilance for molecular epidemiological surveillance of CR-ECC due to the presence of epidemic clones with a phylogenetically diverse and wide array of antimicrobial resistance genes in vulnerable populations.

Recommendations on the measurement and use of the alcohol consumption biomarker CDT. A position paper from the IFCC Working Group on CDT standardisation.

BACKGROUND: Carbohydrate deficient transferrin (CDT) is a biomarker for excessive alcohol consumption utilized in clinical and forensic medicine and workplace testing. Previously, many different analytical methods for CDT were used and the measurand varied considerably, making direct comparison of test results difficult. To end this confusion, the IFCC established a working group on CDT standardisation (WG-CDT) which completed its tasks in 2017. METHODS: This IFCC position paper by the WG-CDT summarizes state of the art information about the measurand and the analytical methods and gives concise recommendations for its utilization. RESULTS: The results achieved by the CDT standardisation process led to accuracy improvements in national external quality assessment schemes over the years. A brief review of ROC based comparison studies with the traditional biomarkers (GGT, MCV, ALT and AST) discusses the bias resulting from inadequate study populations. In large groups of the general population the superior diagnostic performance of CDT is confirmed. CONCLUSION: The relationship between alcohol intake versus resulting CDT is discussed as well as the cutoff and measurement uncertainty. Concerning the application in practice, potential pitfalls are considered and recommendations handling both analytical and preanalytical caveats are given. Finally, some examples of serious misunderstandings in publications about CDT are addressed.

A practical key recovery attack on the lightweight WG-5 stream cipher.

WG-5 is a lightweight stream cipher proposed for usage in the resource-constrained devices, e.g., passive RFID tags, industrial controllers, contactless smart cards and sensors. In this paper, a weakness called slide property of WG-5 which has not been discovered in previous works is for the first time explored and analyzed. The result shows that the probability that two related key-IV pairs of WG-5 generate the shifted keystreams can be up to 2-20, which is significantly high compared with an ideal stream cipher that generates the random keystreams. The correctness and accuracy of this theoretical probability is confirmed experimentally. Based on the slide property of WG-5, some key recovery attacks on WG-5 in the related key setting are proposed. The cryptanalytic result shows that the 80-bit secret key of WG-5 can be recovered with a time complexity of 225.615, requiring 6 related keys and 80 keystream bits for each of 224.585 chosen IVs. The experimental result validates our attack and shows that WG-5 can be broken within about 92.054 seconds on a common PC in the related key setting. These results imply that the design of WG-5 is far from optimal and needs to be strengthened to provide enough security for the lightweight constrained applications.

Clothianidin-resistant Anopheles gambiae adult mosquitoes from Yaoundé, Cameroon, display reduced susceptibility to SumiShield® 50WG, a neonicotinoid formulation for indoor residual spraying.

BACKGROUND: Chronic exposure of mosquito larvae to pesticide residues and cross-resistance mechanisms are major drivers of tolerance to insecticides used for vector control. This presents a concern for the efficacy of clothianidin, an agricultural neonicotinoid prequalified for Indoor Residual Spraying (IRS). METHODS: Using standard bioassays, we tested if reduced susceptibility to clothianidin can affect the efficacy of SumiShield® 50WG, one of four new IRS formulations containing clothianidin. We simultaneously monitored susceptibility to clothianidin and to SumiShield 50WG, testing adults of Anopheles gambiae, An. coluzzii and Culex sp sampled from urban, suburban and agricultural areas of Yaoundé, Cameroon. RESULTS: We found that in this geographic area, the level of susceptibility to the active ingredient predicted the efficacy of SumiShield 50WG. This formulation was very potent against populations that reached 100% mortality within 72 h of exposure to a discriminating concentration of clothianidin. By contrast, mortality leveled off at 75.4 ± 3.5% within 7 days of exposure to SumiShield 50WG in An. gambiae adults collected from a farm where the spraying of the two neonicotinoids acetamiprid and imidacloprid for crop protection is likely driving resistance to clothianidin. CONCLUSIONS: Despite the relatively small geographic extend of the study, the findings suggest that cross-resistance may impact the efficacy of some new IRS formulations and that alternative compounds could be prioritized in areas where neonicotinoid resistance is emerging.

Size compensation in Drosophila after generalised cell death.

Regeneration is a response mechanism aimed to restore tissues that have been damaged. We are studying in the wing disc of Drosophila the regenerative response to a dose of Ionizing Radiation that kills over 35% of the cells distributed all over the disc. After such treatment the discs are able to restore normal size, indicating there is a mechanism that repairs generalised damage. We have tested the role of the JNK, JAK/STAT and Wg pathways, known to be required for regeneration after localised damage in the disc. We find that after irradiation there is size compensation in the absence of function of these pathways, indicating that they are not necessary for the compensation. Furthermore, we also find that generalised damage does not cause an increase in the proliferation rate of surviving cells. We propose that irradiated discs suffer a developmental delay and resume growth at normal rate until they reach the final stereotyped size. The delay appears to be associated with a developmental reversion, because discs undergo rejuvenation towards an earlier developmental stage. We argue that the response to generalized damage is fundamentally different from that to localized damage, which requires activity of JNK and Wg.

The Potential of Soluble Proteins in High-Moisture Soy Protein-Gluten Extrudates Preparation.

In this study, the effects of different soluble proteins, including collagen peptides (CP), soy protein hydrolysate (HSPI), whey protein isolate (WPI), sodium caseinate (SC), and egg white protein (EWP), on the structural and mechanical properties of blends containing soy protein isolate (SPI) and wheat gluten (WG) were investigated using high-moisture extrusion. The addition of CP and HSPI resulted in a more pronounced fibrous structure with increased voids, attributing to their plasticizing effect that enhanced polymer chain mobility and reduced viscosity. WPI, SC, and EWP acted as crosslinking agents, causing early crosslink formation and decreased polymer chain mobility. These structural variations directly influenced the tensile properties of the extrudates, with CP displaying the highest anisotropic index. Moreover, the presence of soluble proteins impacts the permeability of the extrudates. These insights shed light on how soluble proteins can be used to modify the properties of SPI-WG blends, making them suitable for meat analogue production.

Paradoxical behaviour of rainfall and temperature over ecologically sensitive areas along the Western Ghats.

Initial reports signify some specific isolated locations in different latitudes, revealing a paradoxical increase in both heavy and very heavy rainfall events and also an increment in total, i.e., in both rainfall and temperature, over ecologically sensitive areas along the Western Ghats (WG). This paper presents a coherent study of the full-scale of daily rainfall and temperature over 27 well-spaced stations in the study area to determine its extent and investigate whether or not this contradictory behaviour is real. Also, an attempt has been made to assess the differential behaviour of rainfall, temperature, and heavy rainfall events in association with land use and land cover change (LULC). The analysis revealed that rainfall and temperature over the study area are increasing, whereas heavy rainfall events have increased during 1981-2020 with strong peaks after 2000 around 18-19°N (Mumbai metropolitan region), 14-16°N (mining and quarrying regions in Goa), and 9-12°N (a narrow strip of land spanning across the coastal towns of Karnataka and Kerala) latitudes. The majority of the rainfall excess years coincided with El Nino years, indicating that El Nino does not affect rainfall negatively. However, rainfall over the WG is influenced by local relief and cascading topography. The spatial pattern of average annual rainfall shows a decreasing trend from south to north because the elevation and span of rainfall occurrence are higher in the southern part of WG. The findings of the current research will help in building a strategy to address trends and patterns of climatic variables in association with LULC.

SNP heterozygosity, relatedness and inbreeding of whole genomes from the isolated population of the Faroe Islands.

BACKGROUND: The population of the Faroe Islands is an isolated population but very little is known about it from whole genome sequencing. The population of about 50000 people has a high incidence of rare diseases e.g., 1:300 for Primary Carnitine Deficiency. A screening programme was implemented, and eleven persons were also whole genome sequenced at x37 coverage for diagnostic purposes of those cases that were not affected by the known mutations. The purpose of our study is to utilize the high coverage data to explore the genomic variation and the ancestral history of the population. We study the SNP heterozygosity, the pairwise relatedness from kinship, the inbreeding from runs of homozygosity ROH, and we find the minor allele frequency distribution. We estimate the population ancestry and the timing of the founding event by using the whole genomes from eight consenting individuals. RESULTS: We find the number of SNPs and the heterozygosity for the eight individual samples, and for merged samples, for which we also study the relatedness. We find close relatedness between the supposedly unrelated individuals. From ROH, we interpret the high relatedness as an ancient property of the isolated population. A bottleneck event is estimated starting between years [Formula: see text] with a maximum consanguineous population in year [Formula: see text] and similarly consanguineous between years [Formula: see text]. The ancestry analysis shows the population descends from founders of [Formula: see text] European and [Formula: see text] Admixed American ancestry. A distinct clustering near the central European and British populations of the 1000 Genome Project is likely the result of the population isolation and genetic drift. The minor allele frequency distribution suggests many rare variants. CONCLUSIONS: The ancestry is mainly European while the inbreeding is higher compared to European populations and population isolates. The Faroese population has inbreeding more like ancient Europeans. We discovered a bottlenecked and consanguineous population event and estimated it starting in the 1st-4th century as compared to the oldest archaeological findings from the 4th-6th century.

Transcriptional pausing factor M1BP regulates cellular homeostasis by suppressing autophagy and apoptosis in Drosophila eye.

During organogenesis cellular homeostasis plays a crucial role in patterning and growth. The role of promoter proximal pausing of RNA polymerase II, which regulates transcription of several developmental genes by GAGA factor or Motif 1 Binding Protein (M1BP), has not been fully understood in cellular homeostasis. Earlier, we reported that M1BP, a functional homolog of ZKSCAN3, regulates wingless and caspase-dependent cell death (apoptosis) in the Drosophila eye. Further, blocking apoptosis does not fully rescue the M1BPRNAi phenotype of reduced eye. Therefore, we looked for other possible mechanism(s). In a forward genetic screen, members of the Jun-amino-terminal-(NH2)-Kinase (JNK) pathway were identified. Downregulation of M1BP ectopically induces JNK, a pro-death pathway known to activate both apoptosis and caspase-independent (autophagy) cell death. Activation of JNK pathway components can enhance M1BPRNAi phenotype and vice-versa. Downregulation of M1BP ectopically induced JNK signaling, which leads to apoptosis and autophagy. Apoptosis and autophagy are regulated independently by their genetic circuitry. Here, we found that blocking either apoptosis or autophagy alone rescues the reduced eye phenotype of M1BP downregulation; whereas, blocking both apoptosis and autophagy together significantly rescues the M1BP reduced eye phenotype to near wild-type in nearly 85% progeny. This data suggests that the cellular homeostasis response demonstrated by two independent cell death mechanisms, apoptosis and autophagy, can be regulated by a common transcriptional pausing mechanism orchestrated by M1BP. Since these fundamental processes are conserved in higher organisms, this novel functional link between M1BP and regulation of both apoptosis and autophagy can be extrapolated to humans.

The Wnt pathway regulates wing morph determination in Acyrthosiphon pisum.

Wing dimorphism occurs in insects as a survival strategy to adapt to environmental changes. In response to environmental cues, mother aphids transmit signals to their offspring, and the offspring either emerge as winged adults or develop as wingless adults with degeneration of the wing primordia in the early instar stage. However, how the wing morph is determined in the early instar stage is still unclear. Here, we established a surgical sampling method to obtain precise wing primordium tissues for transcriptome analysis. We identified Wnt as a regulator of wing determination in the early second instar stage in the pea aphid. Inhibiting Wnt signaling via knockdown of Wnt2, Wnt11b, the Wnt receptor-encoding gene fz2 or the downstream targets vg and omb resulted in a decreased proportion of winged aphids. Activation of Wnt signaling via knockdown of miR-8, an inhibitor of the Wnt/Wg pathway, led to an increased proportion of winged aphids. Furthermore, the wing primordia of wingless nymphs underwent apoptosis in the early second instar, and cell death was activated by knockdown of fz2 under the wing-inducing condition. These results indicate that the developmental plasticity of aphid wings is modulated by the intrinsic Wnt pathway in response to environmental challenges.

Genomic and Temporal Analysis of Deletions Correlated to qRT-PCR Dropout in N Gene in Alpha, Delta and Omicron Variants.

Since the first SARS-CoV-2 outbreak, mutations such as single nucleotide polymorphisms (SNPs) and insertion/deletions (INDELs) have changed and characterized the viral genome sequence, structure and protein folding leading to the onset of new variants. The presence of those alterations challenges not only the clinical field but also the diagnostic demand due to failures in gene detection or incompleteness of polymerase chain reaction (PCR) results. In particular, the analysis of understudied genes such as N and the investigation through whole-genome next generation sequencing (WG-NGS) of regions more prone to mutate can help in the identification of new or reacquired mutations, with the aim of designing robust and long-lasting primers. In 48 samples of SARS-CoV-2 (including Alpha, Delta and Omicron variants), a lack of N gene amplification was observed in the genomes analyzed through WG-NGS. Three gene regions were detected hosting the highest number of SNPs and INDELs. In several cases, the latter can interfere deeply with both the sensitivity of diagnostic methodologies and the final protein folding. The monitoring over time of the viral evolution and the reacquisition among different variants of the same mutations or different alterations within the same genomic positions can be relevant to avoid unnecessary consumption of resources.