Nesobasis rito sp. nov. (Zygoptera: Coenagrionidae), a new species of forest damselfly from Vanua Levu, Fiji

Nesobasis rito sp. nov. (Holotype ♂, Fiji, Vanua Levu, Drawa, 31 v 2018, A. Rivas-Torres leg.) from the comosa group is here described, illustrated, diagnosed, and compared with morphologically close species of the genus. Nesobasis rito can be distinguished from its related congeners by the shape of the caudal appendages and the ligula. The most similar species are N. comosa and N. heteroneura, which, like N. rito, have the caudal appendages covered by dense setae (especially the first species), but the shape differs clearly in lateral view, with N. rito having longer and more slender appendages, and a basal tooth clearly seen in dorsal view, absent in other members of the comosa group. The specific status of the collected specimens is also supported by the results of genetic analyses, where N. rito appears as a well-supported monophyletic clade. Nesobasis rito also has a distinct distribution from its most similar congeners: it is found on Vanua Levu, while N. comosa is found on Viti Levu and the closely related N. heteroneura is found on Viti Levu and Ovalau. All species of this group are found in streams with native forest riparian vegetation on their respective islands.

iDEP-assisted isolation of insulin secretory vesicles

ABSTRACTOrganelle heterogeneity and inter-organelle associations within a single cell contribute to the limited sensitivity of current organelle separation techniques, thus hindering organelle subpopulation characterization. Here we use direct current insulator-based dielectrophoresis (DC-iDEP) as an unbiased separation method and demonstrate its capability by identifying distinct distribution patterns of insulin vesicles from pancreatic β-cells. A multiple voltage DC-iDEP strategy with increased range and sensitivity has been applied, and a differentiation factor (ratio of electrokinetic to dielectrophoretic mobility) has been used to characterize features of insulin vesicle distribution patterns. We observed a significant difference in the distribution pattern of insulin vesicles isolated from glucose-stimulated cells relative to unstimulated cells, in accordance with functional maturation of vesicles upon glucose stimulation, and interpret this to be indicative of high-resolution separation of vesicle subpopulation. DC-iDEP provides a path for future characterization of subtle biochemical differences of organelle subpopulations within any biological system.

The Impact of Host Genotype, Intestinal Sites and Probiotics Supplementation on the Gut Microbiota Composition and Diversity in Sheep

Three sampling strategies with a 16s rRNA high-throughput sequencing and gene expression assay (by RT-PCR) were designed, to better understand the host and probiotics effect on gut microbiota in sheep. Sampling: (1) colon contents and back-fat tissues from small-tailed Han sheep (SHS), big-tailed Hulun Buir sheep (BHBS), and short-tailed Steppe sheep (SHBS) (n = 12, 14, 12); (2) jejunum, cecum and colon contents, and feces from Tan sheep (TS, n = 6); (3) feces from TS at 4 time points (nonfeeding, 30 and 60 feeding days, and stop feeding 30 days) with probiotics supplementation (n = 7). The results indicated SHS had the highest Firmicutes abundance, the thinnest back-fat, and the lowest expression of C/EBPβ, C/EBPδ, ATGL, CFD, and SREBP1. Some bacteria orders and families could be potential biomarkers for sheep breeds with a distinct distribution of bacterial abundance, implying the host genotype is predominant in shaping unique microbiota under a shared environment. The microbiota diversity and Bifidobacterial populations significantly changed after 60 days of feeding but restored to its initial state, with mostly colonies, after 30 days ceased. The microbiota composition was greatly different between the small and large intestines, but somewhat different between the large intestine and feces; feces may be reliable for studying large intestinal microbiota in ruminants.

Astrocytic IP3Rs: Beyond IP3R2

Astrocytes are sensitive to ongoing neuronal/network activities and, accordingly, regulate neuronal functions (synaptic transmission, synaptic plasticity, behavior, etc.) by the context-dependent release of several gliotransmitters (e.g., glutamate, glycine, D-serine, ATP). To sense diverse input, astrocytes express a plethora of G-protein coupled receptors, which couple, via Gi/o and Gq, to the intracellular Ca2+ release channel IP3-receptor (IP3R). Indeed, manipulating astrocytic IP3R-Ca2+ signaling is highly consequential at the network and behavioral level: Depleting IP3R subtype 2 (IP3R2) results in reduced GPCR-Ca2+ signaling and impaired synaptic plasticity; enhancing IP3R-Ca2+ signaling affects cognitive functions such as learning and memory, sleep, and mood. However, as a result of discrepancies in the literature, the role of GPCR-IP3R-Ca2+ signaling, especially under physiological conditions, remains inconclusive. One primary reason for this could be that IP3R2 has been used to represent all astrocytic IP3Rs, including IP3R1 and IP3R3. Indeed, IP3R1 and IP3R3 are unique Ca2+ channels in their own right; they have unique biophysical properties, often display distinct distribution, and are differentially regulated. As a result, they mediate different physiological roles to IP3R2. Thus, these additional channels promise to enrich the diversity of spatiotemporal Ca2+ dynamics and provide unique opportunities for integrating neuronal input and modulating astrocyte–neuron communication. The current review weighs evidence supporting the existence of multiple astrocytic-IP3R isoforms, summarizes distinct sub-type specific properties that shape spatiotemporal Ca2+ dynamics. We also discuss existing experimental tools and future refinements to better recapitulate the endogenous activities of each IP3R isoform.

Endophytic Fungi of Emersed Halophytes in River Deltas and Tidal Flats of the Korean Ramsar Wetlands

This study aimed to obtain information on the diversity and distribution of the endophytic fungi in Ramsar wetlands. Vast salt marshes in Suncheon Bay, Korea, are formed by two types of ecotones (tidal flats and deltas) that are supported by the emersed halophytes Phragmites australis and Suaeda japonica. Overall, 324 endophytes were isolated from P. australis (six sampling points in the delta and five in the tidal flats) and S. japonica (six in tidal flats). Margalef’s, Menhinick’s, Shannon’s, and Simpson’s diversity indices significantly varied among the ecotones. In particular, higher variance in diversity value and unevenness was observed in the delta marsh compared with the tidal flat marsh. Further, morphological diversity in the delta salt marsh was 1.8 times higher than that of the tidal flat. Comprising several dominant genera (Aspergillus, Cladosporium, and Penicillium), Epicoccum, Paraconiothyrium, Septoriella, and Talaromyces were widely distributed regardless of the aquatic conditions or halophyte species. This study highlights the distinct distribution and diversity of marine endophytes in various ecotones that can physically protect the coastal areas. This data is of much value to secure a national culture collection for future restoration of the coastal wetlands and their ecosystems.

Analysis of the Regulatory Science Applied to a Single Portfolio of Eight Biosimilar Product Approvals by Four Key Regulatory Authorities

Slow uptake of biosimilars in some regions is often attributed to a lack of knowledge combined with concerns about safety and efficacy. To alleviate physician and patient apprehensions, regulatory reviews from four major regulatory authorities (RAs) (European Medicines Agency, US Food and Drug Administration, Health Canada, and Japan Pharmaceuticals and Medical Devices Authority) across a portfolio of eight biosimilars were analyzed to provide insight into RA review focus and approach. RA queries were evaluated in an unbiased and systematic manner by major classification (Chemistry, Manufacturing and Controls [CMC], nonclinical, clinical or regulatory) and then via detailed sub-classification. There was a consistent, predominant focus on CMC from all RAs. The review focus based on sub-classification of clinical and regulatory queries was influenced by molecular complexity, with significant differences between categories (monoclonal antibody or protein) in the distribution of query topics; specifically, bioanalytical (p = 0.023), comparative safety and efficacy (p = 0.023), and statutory (including the justification of extrapolation) (p = 0.00033). Each biosimilar had a distinct distribution of clinical query topics, tailored to product-specific data. This analysis elucidated areas of heightened RA interest, and validated their application of regulatory science in the evaluation of biosimilar safety and efficacy.

Cargo sorting zones in the trans-Golgi network visualized by super-resolution confocal live imaging microscopy in plants

The trans-Golgi network (TGN) has been known as a key platform to sort and transport proteins to their final destinations in post-Golgi membrane trafficking. However, how the TGN sorts proteins with different destinies still remains elusive. Here, we examined 3D localization and 4D dynamics of TGN-localized proteins of Arabidopsis thaliana that are involved in either secretory or vacuolar trafficking from the TGN, by a multicolor high-speed and high-resolution spinning-disk confocal microscopy approach that we developed. We demonstrate that TGN-localized proteins exhibit spatially and temporally distinct distribution. VAMP721 (R-SNARE), AP (adaptor protein complex)−1, and clathrin which are involved in secretory trafficking compose an exclusive subregion, whereas VAMP727 (R-SNARE) and AP-4 involved in vacuolar trafficking compose another subregion on the same TGN. Based on these findings, we propose that the single TGN has at least two subregions, or “zones”, responsible for distinct cargo sorting: the secretory-trafficking zone and the vacuolar-trafficking zone.

Ancient Origin of Two 5S rDNA Families Dominating in the Genus Rosa and Their Behavior in the Canina-Type Meiosis

The genus Rosa comprises more than 100 woody species characterized by intensive hybridization, introgression, and an overall complex evolutionary history. Besides many diploid species (2n = 2x = 14) polyploids ranging from 3x to 10x are frequently found. Here we analyzed 5S ribosomal DNA in 19 species covering two subgenera and the major sections within subg. Rosa. In addition to diploids and polyploids with regular meiosis, we focused on 5x dogroses (Rosa sect. Caninae), which exhibit an asymmetric meiosis differentiating between bivalent- and univalent-forming chromosomes. Using genomic resources, we reconstructed 5S rDNA units to reveal their phylogenetic relationships. Additionally, we designed locus-specific probes derived from intergenic spacers (IGSs) and determined the position and number of 5S rDNA families on chromosomes. Two major 5S rDNA families (termed 5S_A and 5S_B, respectively) were found at variable ratios in both diploid and polyploid species including members of the early diverging subgenera, Rosa persica and Rosa minutifolia. Within subg. Rosa species of sect. Rosa amplified the 5S_A variant only, while taxa of other sections contained both variants at variable ratios. The 5S_B family was often co-localized with 35S rDNA at the nucleolar organizer regions (NOR) chromosomes, whereas the co-localization of the 5S_A family with NOR was only exceptionally observed. The allo-pentaploid dogroses showed a distinct distribution of 5S rDNA families between bivalent- and univalent-forming chromosomes. In conclusion, two divergent 5S rDNA families dominate rose genomes. Both gene families apparently arose in the early history of the genus, already 30 myrs ago, and apparently survived numerous speciation events thereafter. These observations are consistent with a relatively slow genome turnover in the Rosa genus.

Time series analysis of the demand for COVID-19 related chest imaging during the first wave of the SARS-CoV-2 pandemic: An explorative study

Objectives The aim of this study was to investigate possible patterns of demand for chest imaging during the first wave of the SARS-CoV-2 pandemic and derive a decision aid for the allocation of resources in future pandemic challenges. Materials and methods Time data of requests for patients with suspected or confirmed coronavirus disease 2019 (COVID-19) lung disease were analyzed between February 27th and May 27th 2020. A multinomial logistic regression model was used to evaluate differences in the number of requests between 3 time intervals (I1: 6am - 2pm, I2: 2pm - 10pm, I3: 10pm - 6am). A cosinor model was applied to investigate the demand per hour. Requests per day were compared to the number of regional COVID-19 cases. Results 551 COVID-19 related chest imagings (32.8% outpatients, 67.2% in-patients) of 243 patients were conducted (33.3% female, 66.7% male, mean age 60 ± 17 years). Most exams for outpatients were required during I2 (I1 vs. I2: odds ratio (OR) = 0.73, 95% confidence interval (CI) 0.62–0.86, p = 0.01; I2 vs. I3: OR = 1.24, 95% CI 1.04–1.48, p = 0.03) with an acrophase at 7:29 pm. Requests for in-patients decreased from I1 to I3 (I1 vs. I2: OR = 1.24, 95% CI 1.09–1.41, p = 0.01; I2 vs. I3: OR = 1.16, 95% CI 1.05–1.28, p = 0.01) with an acrophase at 12:51 pm. The number of requests per day for outpatients developed similarly to regional cases while demand for in-patients increased later and persisted longer. Conclusions The demand for COVID-19 related chest imaging displayed distinct distribution patterns depending on the sector of patient care and point of time during the SARS-CoV-2 pandemic. These patterns should be considered in the allocation of resources in future pandemic challenges with similar disease characteristics.

Updated SARS-CoV-2 Single Nucleotide Variants and Mortality Association

Since its outbreak in December 2019, COVID-19 has caused 100,5844,555 cases and 2,167,313 deaths as of Jan 27, 2021. Comparing our previous study of SARS-CoV-2 single nucleotide variants (SNVs) before June 2020, we found out that the SNV clustering had changed considerably since June 2020. Apart from that the group SNVs represented by two non-synonymous mutations A23403G (S: D614G) and C14408T (ORF1ab: P4715L) became dominant and carried by over 95% genomes, a few emerging groups of SNVs were recognized with sharply increased monthly occurrence ratios up to 70% in November 2020. Further investigation revealed that several SNVs were strongly associated with the mortality, but they presented distinct distribution in specific countries, e.g., Brazil, USA, Saudi Arabia, India, and Italy. SNVs including G25088T, T25A, G29861T and G29864A were adopted in a regularized logistic regression model to predict the mortality status in Brazil with the AUC of 0.84. Protein structure analysis showed that the emerging subgroups of non-synonymous SNVs and those mortality-related ones in Brazil were located on protein surface area. The clashes in protein structure introduced by these mutations might in turn affect virus pathogenesis through conformation changes, leading to the difference in transmission and virulence. Particularly, we found that SNVs tended to occur in intrinsic disordered regions (IDRs) of Spike (S) and ORF1ab, suggesting a critical role of SNVs in protein IDRs to determine protein folding and immune evasion.