Miracle-dz/UniFAR-FTUs huggingface dataset:
Ninjin'yoeito Targets Distinct Ca2+ Channels to Activate Ghrelin-Responsive vs. Unresponsive NPY Neurons in the Arcuate Nucleus

Appetite loss or anorexia substantially deteriorates quality of life in various diseases, and stand upstream of frailty. Neuropeptide Y (NPY) in the hypothalamic arcuate nucleus (ARC) and ghrelin released from stomach are potent inducers of appetite. We previously reported that Ninjin'yoeito, a Japanese kampo medicine comprising twelve herbs, restores food intake, and body weight in cisplatin-treated anorectic mice. Furthermore, Ninjin'yoeito increased cytosolic Ca2+ concentration ([Ca2+]i) in not only ghrelin-responsive but ghrelin-unresponsive NPY neurons in ARC. The cellular lineage/differentiation of ghrelin-unresponsive neuron is less defined but might alter along with aging and diet. This study examined the occupancy of ghrelin-unresponsive neurons among ARC NPY neurons in adult mice fed normal chow, and explored the mechanisms underlying Ninjin'yoeito-induced [Ca2+]i increases in ghrelin-unresponsive vs. ghrelin-responsive NPY neurons. Single ARC neurons were subjected to [Ca2+]i measurement and subsequent immunostaining for NPY. Ghrelin failed to increase [Ca2+]i in 42% of ARC NPY neurons. Ninjin'yoeito (10 μg/ml)-induced increases in [Ca2+]i were abolished in Ca2+ free condition in ghrelin-responsive and ghrelin-unresponsive ARC NPY neurons. Ninjin'yoeito-induced [Ca2+]i increases were inhibited by N-type Ca2+ channel blocker ω-conotoxin in the majority (17 of 20), while by L-type Ca2+ channel blocker nitrendipine in the minority (2 of 23), of ghrelin-responsive neurons. In contrast, Ninjin'yoeito-induced [Ca2+]i increases were inhibited by nitrendipine in the majority (14 of 17), while by ω-conotoxin in the minority (8 of 24), of ghrelin-unresponsive neurons. These results indicate that ghrelin-unresponsive neurons occur substantially among NPY neurons of ARC in adult mice fed normal chow. Ninjin'yoeito preferentially target N-type and L-type Ca2+ channels in the majority of ghrelin-responsive and ghrelin-unresponsive neurons, respectively, to increase [Ca2+]i. We suggest ARC N- and L-type Ca2+ channels as potential targets for activating, respectively, ghrelin-responsive, and unresponsive NPY neurons to treat anorexia.
Evaluation of NGS-based approaches for SARS-CoV-2 whole genome characterisation

Since the beginning of the COVID-19 outbreak, SARS-CoV-2 whole-genome sequencing (WGS) has been performed at unprecedented rate worldwide with the use of very diverse Next Generation Sequencing (NGS) methods. Herein, we compare the performance of four NGS-based approaches for SARS-CoV-2 WGS. Twenty four clinical respiratory samples with a large scale of Ct values (from 10.7 to 33.9) were sequenced with four methods. Three used Illumina sequencing: an in-house metagenomic NGS (mNGS) protocol and two newly commercialized kits including a hybridization capture method developed by Illumina (DNA Prep with Enrichment kit and Respiratory Virus Oligo Panel, RVOP) and an amplicon sequencing method developed by Paragon Genomics (CleanPlex SARS-CoV-2 kit). We also evaluated the widely used amplicon sequencing protocol developed by ARTIC Network and combined with Oxford Nanopore Technologies (ONT) sequencing. All four methods yielded near-complete genomes (>99%) for high viral loads samples, with mNGS and RVOP producing the most complete genomes. For mid viral loads, 2/8 and 1/8 genomes were incomplete (<99%) with mNGS and both CleanPlex and RVOP, respectively. For low viral loads (Ct ≥25), amplicon-based enrichment methods were the most sensitive techniques yielding complete genomes for 7/8 samples. All methods were highly concordant in terms of identity in complete consensus sequence. Just one mismatch in two samples was observed in CleanPlex vs the other methods, due to the dedicated bioinformatics pipeline setting a high threshold to call SNP compared to reference sequence. Importantly, all methods correctly identified a newly observed 34-nt deletion in ORF6 but required specific bioinformatic validation for RVOP. Finally, as a major warning for targeted techniques, a default of coverage in any given region of the genome should alert to a potential rearrangement or a SNP in primer annealing or probe-hybridizing regions and would require regular updates of the technique according to SARS-CoV-2 evolution.
Cortical Presynaptic Boutons Progressively Engulf Spinules as They Mature

Despite decades of discussion in the neuroanatomical literature, the role of the synaptic ‘spinule’ in synaptic development and function remains elusive. Canonically, spinules are finger-like projections that emerge from postsynaptic spines and can become enveloped by presynaptic boutons. When a presynaptic bouton encapsulates a spinule in this manner, the membrane apposition between the spinule and surrounding bouton can be significantly larger than the membrane interface at the synaptic active zone. Hence, spinules may represent a mechanism for extrasynaptic neuronal communication, and/or may function as structural ‘anchors’ that increase the stability of cortical synapses. Yet despite their potential to impact synaptic function, we have little information on the percentages of developing and adult cortical bouton populations that contain spinules, the percentages of these cortical spinule-bearing boutons (SBBs) that contain spinules from distinct neuronal/glial origins, or whether the onset of activity or cortical plasticity are correlated with increased prevalence of cortical SBBs. Here, we employed 2D and 3D electron microscopy to determine the prevalence of spinules in excitatory presynaptic boutons at key developmental time points in the primary visual cortex (V1) of female and male ferrets. We find that the prevalence of SBBs in V1 increases across postnatal development, such that ~25% of excitatory boutons in late adolescent ferret V1 contain spinules. In addition, we find that a majority of spinules within SBBs at later developmental time points emerge from postsynaptic spines and adjacent boutons/axons, suggesting that synaptic spinules may enhance synaptic stability and allow for axo-axonal communication in mature sensory cortex. Significance Statement Synaptic spinules are finger-like projections from neurites that can become completely embedded within presynaptic boutons, potentially enhancing synaptic communication and stability. Yet while their existence has been discussed for decades, spinule prevalence, projection origins, and relationship to neocortical sensory activity remain unknown. In this study, we employed 2D and 3D electron microscopy techniques to characterize the development of excitatory cortical spinule-bearing boutons (SBBs) and their relationship to sensory activity and plasticity. Our results demonstrate that neocortical presynaptic SBB prevalence is not correlated with the onset of sensory activity or heightened cortical plasticity, and that by late adolescence nearly one-quarter of presynaptic boutons contain a spinule. Hence, synaptic spinules may play progressively important roles in cortical function as these synapses mature.
Does Customer Demotion Jeopardize Loyalty?

Hierarchical loyalty programs award elevated customer status (e.g., “elite membership”) to consumers who meet a predefined spending level. However, if a customer subsequently falls short of the required spending level, firms commonly revoke that status. The authors investigate the impact of such customer demotion on loyalty intentions toward the firm. Building on prospect theory and emotions theory, the authors hypothesize that changes in customer status have an asymmetric negative effect, such that the negative impact of customer demotion is stronger than the positive impact of status increases. An experimental scenario study provides evidence that loyalty intentions are indeed lower for demoted customers than for those who have never been awarded a preferred status, meaning that hierarchical loyalty programs can drive otherwise loyal customers away from a firm. A field study using proprietary sales data from a different industry context demonstrates the robustness of the negative impact of customer demotion. The authors test the extent to which design variables of hierarchical loyalty programs may attenuate the negative consequences of status demotions with a second experimental scenario study and present an analytical model that links status demotion to customer equity to aid managerial decision making.
Reconstitution of human kinetochore in mitotic cell extracts reveals permitted and restricted assembly steps

Assembly of a functional kinetochore is critical for accurate chromosome segregation. Hierarchical recruitment of soluble components during kinetochore assembly is a highly regulated mitotic event, but the underlying steps are not well understood. In yeast and Xenopus egg extracts, soluble kinetochore components can spontaneously assemble into microtubule-binding subcomplexes. Although the molecular interactions among specific kinetochore components are evolutionary conserved in eukaryotes, it remains unclear which de novo assembly steps are permitted in extracts of mitotic human cells. By analyzing the recruitment of GFP-fused kinetochore proteins from human mitotic cell extracts to inner kinetochore components immobilized on microbeads, we reconstructed the interaction between CENP-C and CENP-A–containing nucleosomes. However, subsequent phospho-dependent binding of the Mis12 complex was less efficient, whereas binding of the Ndc80 complex was inhibited. Consistently, the microtubule-binding activity of native kinetochore components, as well as those assembled using a combination of native and recombinant human proteins, was weaker than that of recombinant Ndc80 complex alone. Such inhibitory mechanisms that prevent interactions between different kinetochore components are likely to guard against spurious formation of kinetochores in the cytosol of mitotic human cells, and imply existence of specific regulatory mechanisms that permit these interactions at the assembling kinetochore.
Safety evaluation of Morinda citrifolia (noni) leaves extract: assessment of genotoxicity, oral short term and subchronic toxicity

Morinda citrifolia L (noni) is an evergreen or small tree that grows in many tropical regions of the world. The use of the noni leaves has not been so studied however; there are reports of its pharmacological benefits. Aims: The objective of this investigation was to assess the genotoxicity, short-term, and subchronic oral toxicity of Morinda citrifolia L leaves aqueous extract. Methods: The genotoxicity of the M. citrifolia extract was investigated by measuring the frequency of micronuclei in mice bone marrow cells. The animals were treated with three doses of the extract (500, 1000, and 2000 mg/kg). For short-term toxicity, both sexes Wistar rats received 1000 mg/kg/day for 28 days. Animals were sacrificed for hematological and biochemical evaluation. For the subchronic study, Wistar rats were administered with three doses of M. citrifolia extract (100, 300, and 1000 mg/kg) by oral route for 90 days. Mortalities, clinical signs, body weight changes, food and water consumption, hematological and biochemical parameters, gross findings, organ weights, and histological examination were monitored during the study period. Results: Genotoxicity and short-term toxicity test resulted in absence of toxicity at doses between 500 and 2000 mg/kg. Significant differences were observed in hemoglobin, and differential leukocyte count after subchronic dosing of the extract. Histology evaluation did not reveal treatment-related abnormalities. Variations observed were within to normal range and reversible. Conclusions: In summary, 1000 mg/kg orally was the NOAEL for M. citrifolia extract for effects other than transient variations in some hematological parameters within normal range.
Superparamagnetic iron oxide: a novel tracer for sentinel lymph node detection in vulvar cancer

Objectives Superparamagnetic techniques for sentinel lymph node (SLNs) biopsy in breast cancer is well recognized but remains novel in the literature in relation to early stage vulvar cancer. The aim of this study was to compare and validate SLN detection using a superparamagnetic iron oxide tracer and a magnetometer probe compared with the standard procedure with a radioisotope (99Tc–technetium 99) and a gamma probe, in patients with vulvar cancer. Methods Patients were included in the study with squamous vulvar tumors less than 4 cm in diameter and without suspicious groin lymph nodes on preoperative magnetic resonance imaging. Patients must have previously qualified for SLN biopsy with a radiotracer as the standard of care. The primary endpoint was the proportion of successful SLN detection with superparamagnetic iron oxide tracer versus 99Tc. The secondary endpoints were average number of SLNs retrieved per patient, proportion of SLNs detected (nodal detection rate), and proportion of pathologically positive results (malignancy rate) per patient and per node comparing both SLN detection methods. Results A total of 20 patients were included in the study. SLNs were found in all patients with both methods, resulting in similar average distributions (3.1/3.2 SLN per patient). The SLN detection rate per patient was 100% with both techniques. Nodal detection sensitivity was 98.5% for the superparamagnetic technique and 93.8% for the radiotracer. Percentage of metastatic lymph nodes detected was 100% with both tracers. The rate of lymph node positivity was 21.5% (14 lymph nodes with metastases) and for patients 45% (9 patients with nodal metastases). Additionally, SLN tainted brown due to superparamagnetic iron oxide nanoparticles in 19 of 20 patients. Conclusions The use of superparamagnetic iron oxide tracer in patients with vulvar cancer seems reliable and not inferior to the standard approach with radiotracer.
Overexpression of HLA class I predominantly on alpha cell in at risk individuals for type 1 diabetes

Human leukocyte antigens of class-I (HLA-I) molecules are hyper-expressed in insulin-containing islets (ICI) of type 1 diabetic (T1D) donors. This study investigated the HLA-I expression in autoantibody positive (AAB+) donors and more closely defined its intra-islet and intracellular localization as well as proximity to infiltrating CD8 T cells with high-resolution confocal microscopy. We found HLA-I hyper-expression had already occurred prior to clinical diagnosis of T1D in islets of AAB+ donors. Interestingly, throughout all stages of disease, HLA-I was mostly expressed by alpha cells. Hyper-expression in AAB+ and T1D donors was associated with intra-cellular accumulation in the Golgi. Proximity analysis showed a moderate but significant correlation between HLA-I and infiltrating CD8 T cells only in ICI of T1D donors, but not in AAB+ donors. These observations not only demonstrate a very early, islet-intrinsic immune-independent increase of HLA-I during diabetes pathogenesis, but also point towards a role for alpha cells in T1D. Summary Human leukocyte antigens of class-I (HLA-I) are hyper-expressed in insulin-containing islets of type 1 diabetic (T1D) donors. This study finds HLA-I hyper-expression occurs prior to clinical diagnosis of T1D in islets of autoantibody-positive donors and mostly expressed by alpha cells.
A SUBSTANTIAL ENERGETIC COST TO MALE REPRODUCTION IN A SEXUALLY DIMORPHIC UNGULATE

Whereas the energetic costs of reproduction for female ungulates are well documented, those of males are not. We investigated age- and sex-specific changes in the kidney fat reserves of a large sample of Himalayan tahr (Hemitragus jemlahicus), a highly sexually dimorphic and polygynous mountain ungulate, during and following the mating period. Subadult male tahr (2–4 years old) obtain copulations opportunistically by “coursing,” whereas adult males (>4 years old) spend more time traveling and searching for females, and obtain copulations after engaging in more time-consuming “tending” and “blocking” displays. If there is a cost to these reproductive behaviors, we predicted that (1) the kidney fat index (KFI) of reproductive males (≥2 years old) should decline during the mating period relative to females and nonreproductive males, and (2) that the KFI of adult males should decline more than that of subadult males. We formulated our predictions into a series of candidate models, including three covariates also likely to affect fat reserves, and compared support for these models using Akaike's Information Criterion (AIC). The best-fitting model revealed that fat reserves in females and nonreproductive males changed similarly: Fat reserves increased during the mating period, in late autumn and early winter, reached a peak in mid-winter, and then declined throughout the remainder of winter and early spring. In contrast, the fat reserves of reproductive males declined during the mating period, as predicted, and remained low throughout winter. The extent of decline in fat reserves was greater in adult males than in subadult males, also as predicted. Our results strongly imply that male tahr incur a substantial energetic cost for engaging in reproduction and that they trade off reproductive success with reduced body condition during winter, which may lower survival.
Choice can be predicted from populations of bursting neurons in superficial layers of monkey V1

Primary visual cortex (V1) is absolutely necessary for normal visual processing, but whether V1 encodes upcoming behavioral decisions based on visual information is an unresolved issue, with conflicting evidence. Further, no study so far has been able to predict choice from time-resolved spiking activity in V1. Here, macaque monkeys are tested in a decision-making task with a categorical variable: match versus non-match of two consecutive stimuli. Our aim is to predict the binary choice “same” versus “different” on trials that only differ in choice, but not in stimuli. Classical decoding schemes fail to predict the choice in such a setting, however, the choice can be predicted by learning the structure of population responses in trials with correct behavior. The learned structure of population responses transfers to the representation of the choice, even when the choice is incorrect. With such generalized learning scheme, the choice can be successfully predicted from spiking activity of neural ensembles in V1 in single trials. We show that the choice signal is primarily carried by bursting neurons in the superficial layer of the cortex. Results suggest that bursting of single neurons and noise correlations between neurons with similar decoding selectivity are dynamical mechanisms that help the formation of time-resolved choice signals. Several lines of evidence point towards the feedback from higher brain areas as the source of information on the upcoming behavioral choice in V1.
A gustatory receptor tuned to the steroid plant hormone brassinolide in Plutella xylostella (Lepidoptera: Plutellidae)

Feeding and oviposition deterrents help phytophagous insects to identify host plants. The taste organs of phytophagous insects contain bitter gustatory receptors (GRs). To explore their function, we focused on PxylGr34, a bitter GR in Plutella xylostella (L.). We detected abundant PxylGr34 transcripts in the larval head and specific expression of PxylGr34 in the antennae of females. Analyses using the Xenopus oocyte expression system and two-electrode voltage-clamp recording showed that PxylGr34 is specifically tuned to the plant hormone brassinolide (BL) and its analog 24-epibrassinolide. Electrophysiological analyses revealed that the medial sensilla styloconica on the maxillary galea of the 4th instar larvae are responsive to BL. Dual-choice bioassays demonstrated that BL inhibits larval feeding and female ovipositing. Knock-down of PxylGr34 by RNAi abolished BL-induced feeding inhibition. These results shed light on gustatory coding mechanisms and deterrence of insect feeding and ovipositing, and may be useful for designing plant hormone-based pest management strategies.
Fish community and bioassessment responses to stream network position

Abstract If organisms move beyond the boundaries of local sampling units, regional metacommunity dynamics could undermine the ability of bioassessment studies to characterize local environmental quality. We tested the prediction that fish dispersal influences local fish community structure and bioassessment metrics as a function of site position within stream networks. We evaluated fish community data from the US Environmental Protection Agency's Regional Environmental Monitoring and Assessment Program in West Virginia, USA, to compare the influences of stream network position, ecoregion, basin, and stream size on local fish community composition. We assigned sites to 1 of 3 stream network positions: 1) main channels (MC, n = 12) encompassed streams with upstream catchment areas >200 km2, 2) mainstem tributaries (MT, n = 43) flowed into MC-sized confluences within 15 fluvial km, 3) headwater tributaries (HT, n = 31) lacked such riverine confluences within 15 fluvial km. MT and HT sites had similar upstream catchment sizes and landuse gradients, but species richness was greater in MT sites than HT sites, whereas MT and MC sites were not different in this regard. Three bioassessment metrics were greater in MT sites than HT sites (intolerant species richness, cyprinid species richness, benthic species richness), but a multimetric index of biotic integrity did not differ among stream network positions. Ordinations revealed that fish community composition was organized primarily by zoogeographic basin (Monongahela River basin, New River basin, Ohio River basin), ecoregion (Central Appalachian Plateau, Western Appalachian Plateau, Ridge and Valley), and stream size. Riverine specialists were more abundant in MT than HT sites and were more abundant in basins connected to the Ohio River than in basins isolated from the Ohio River by a large waterfall (New River). Our results suggest that contemporary dispersal among streams influences fish community composition over small spatial scales (101 km), historical dispersal constrained by zoogeographic barriers influences community structure over larger areas (102 km), and contemporary dispersal by fishes influences certain metrics commonly used in bioassessment programs.
Unexpected diversity of CPR bacteria and nanoarchaea in the rare biosphere of rhizosphere-associated grassland soil

Candidate Phyla Radiation (CPR) bacteria and nanoarchaea populate most ecosystems, but are rarely detected in soil. We concentrated particles less than 0.2 μm from grassland soil, enabling targeted metagenomic analysis of these organisms, which are almost totally unexplored in soil. We recovered a diversity of CPR bacteria and some nanoarchaea sequences, but no sequences from other cellular organisms. The sampled sequences include Doudnabacteria (SM2F11) and Pacearchaeota, organisms not previously reported in soil, as well as Saccharibacteria, Parcubacteria and Microgenomates. CPR and DPANN (an acronym of the names of the first included archaea phyla) enrichments of 100-1000-fold were achieved compared to bulk soil, in which we estimate these organisms comprise about 1 to 100 cells per gram of soil. Like most CPR and DPANN sequenced to date, we predict these microorganisms live symbiotic, anaerobic lifestyles. However, Saccharibacteria, Parcubacteria, and Doudnabacteria genomes sampled here also encode ubiquinol oxidase operons that may have been acquired from other bacteria, likely during adaptation to aerobic soil environments. We posit that although present at low abundance, CPR bacteria and DPANN archaea could impact overall soil microbial community function by modulating host organism abundances and activity.