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Objectives The aim of this AFIRE (Atrial Fibrillation and Ischemic Events With Rivaroxaban in Patients With Stable Coronary Artery Disease) trial subgroup analysis was to examine rivaroxaban monotherapy benefits and their relation to the time between stenting and enrollment among patients after coronary stenting. Background Of 2,215 patients with atrial fibrillation and stable coronary artery disease in the AFIRE trial, rivaroxaban monotherapy was noninferior to rivaroxaban plus antiplatelet therapy (combination therapy) in terms of efficacy and superior for safety endpoints. However, thrombotic risk after antiplatelet therapy cessation remained a concern among 1,444 patients who had undergone coronary stenting >1 year before enrollment. Methods The benefits of rivaroxaban monotherapy in coronary stenting subgroups were assessed for efficacy (a composite of stroke, systemic embolism, myocardial infarction, unstable angina requiring revascularization, or death of any cause), safety (major bleeding defined according to International Society on Thrombosis and Haemostasis criteria), ischemic endpoints, net adverse clinical event, and time between stenting and enrollment. Results Efficacy and safety endpoints for monotherapy were superior to combination therapy, with HRs of 0.70 for efficacy (95% CI: 0.50-0.98; P = 0.036) and 0.55 for safety (95% CI: 0.33-0.92; P = 0.019). For ischemic endpoints, the HR was 0.82 (95% CI: 0.58-1.15; P = 0.240). The HR became smaller with longer time between stenting and enrollment (efficacy, P for interaction = 0.158; safety, P = 0.097). Conclusions In patients with atrial fibrillation after coronary stenting, the benefits of rivaroxaban monotherapy for efficacy and safety endpoints were consistent with those in the whole AFIRE trial population. The benefits became apparent with longer time between stenting and enrollment. (Atrial Fibrillation and Ischemic Events With Rivaroxaban in Patients With Stable Coronary Artery Disease Study [AFIRE]; UMIN000016612 , NCT02642419 )

Male-killing, a male-specific death of arthropod hosts during development, is induced by

The endosymbiotic bacterium Wolbachia occasionally increases host fitness or manipulates host reproductions to enhance vertical transmission. Multiple Wolbachia strains can coinfect the same host individual, which alters the density as well as phenotypes of the bacteria. However, the effects of Wolbachia coinfection on host fitness remain largely unknown. Here, we examined the effects of three phylogenetically distinct Wolbachia strains, wHm-a, wHm-b, and wHm-c, on host fitness by comparing non-infected, singly infected, and triply infected Homona magnanima lines within a fixed genetic background. By examining the effects of Wolbachia on host longevity, survivorship, and reproduction, we demonstrated that single infection with either wHm-b or wHm-c reduced host reproduction, but the triple infection led to the highest intrinsic growth rate. Susceptibility to the natural pathogens such as viruses and fungi was not different among the lines regardless of Wolbachia infection status. Cellular and humoral immunities were not affected by Wolbachia in females, whereas phenoloxidase activity was suppressed in males of all Wolbachia-infected lines, implying that it was a result of the mother's curse hypothesis or a strategy of Wolbachia to increase their horizontal transmission efficiency. Although how the host's genetic diversity affects the Wolbachia fitness effects is yet unknown, our findings indicated that the effects of Wolbachia are deeply influenced by infection status and that Wolbachia could change symbiotic strategy depending on host sex and transmission route.

Protection from parasitism by a virus Parasitoid wasps have developed myriad systems to overcome the defense mechanisms of their hosts as they lay their eggs in the bodies and eggs of targeted species. Gasmi et al . report how the host can fight back when infected by a virus that expresses a protein conferring resistance to the parasitoid. When members of the butterfly and moth family are targeted by wasps, a protein family has evolved that is horizontally carried by viruses—and sometimes is incorporated into the host genome—and impairs the ability of parasitoid offspring to fully develop and emerge. Characterizing the ability of this protein to protect hosts against specific parasites, the authors document an ongoing host-parasite evolutionary arms race. —LMZ

The microsporidian Nosema ceranae commonly infects the Asian honey bee Apis cerana Fabricius (Hymenoptera: Apidae) and poses a high potential risk to co-occurring A. mellifera. Here, we demonstrated the effect of N. ceranae isolated from a wild colony of A. cerana japonica in Tokyo on A. mellifera workers. Sequencing analyses of the Tokyo N. ceranae isolate using the SSU rRNA gene showed a high probability match to the European N. ceranae isolates. Experimental exposure of A. mellifera workers with 102–104 spores of N. ceranae resulted in 100% infection but did not significantly affect mortality, despite spore load doubling over the duration of infection. Nosema ceranae populations are geographically mixed, likely by the worldwide commercial trade of honeybees for commercial pollination. More than 100 years of A. mellifera beekeeping in Japan with repeated importations may promote N. ceranae transmission among A. mellifera and A. cerana japonica populations. Our results did not detect the lethal effects of the N. ceranae infection on A. mellifera workers. However, N. ceranae infections sometimes cause the sublethal effects, such as digestive disruption and nutritional stress, potentially resulting in reduced colony-level productivity. Therefore, we need to monitor the N. ceranae infections on A. mellifera.

ABSTRACT: The time-dependent behavior of salt rocks poses challenges to well design. Salt mobility is associated with critical wellbore closure and excessive casing loads. In the presence of cement imperfections, non-uniform loads on casing strings can lead to failure. A parametric finite-element analysis considers an uncemented annulus, a fully cemented annulus, and a partially cemented annulus with a channel covering 20%, 50% and 80% of the pipe circumference. No pressure build-up was considered. Commonly applied to Brazilian salts, a double-mechanism creep law is adopted, and strain rate uncertainties are accounted for. The stress distribution is less severe when uniform loads act on the casing surface. A partially cemented annulus results in non-uniform loads and deformation of the casing cross-section. A small volume of cement channels leads to localized regions of critical stress within the casing. As the channel size increases, the higher moment arm results in higher casing stresses, especially when two-point bending occurs at the salt-casing contact. Analysis of casing yield and failure when cement channels are present along salt sections allows for more accurate evaluation of casing integrity. 1. INTRODUCTION Well design in salt formations faces many challenges due to the time-dependent behavior of these rocks. Wellbore closure due to salt creep may lead to drill string sticking or aborted casing runs, and the correct drilling mud weight must be applied. After casing is set, salt creep can increase casing loads and compromise well integrity. When submitted to a deviatoric stress, salt exhibits a time-dependent strain response according to Figure 1. During the first stage of loading (primary creep) salt undergoes a high rate of deformation, that gradually slows down until it reaches steady state (Secondary) creep. Finally, during tertiary creep the strain rate increases until rupture (Jaeger et al, 2007; Goodman, 1989). The strain rate is influenced by many factors, such as mineralogical composition, impurities, formation temperature and the imposed differential stress (Liu et al., 2013, Fjaer et al., 2008).

The origin and mechanism of male-killing, an advantageous strategy employed by maternally transmitted symbionts such as Wolbachia, remain unclear. We compared genomes of four Wolbachia strains derived from Homona magnanima, a male-killing strain wHm-t (1.5 Mb), and three non-male-killing strains, wHm-a (1.1 Mb), wHm-b (1.3 Mb), and wHm-c (1.4 Mb). A wHm-t-specific 76-kbp prophage region harboured two tandemly arrayed WO-mediated killing (wmk) gene homologs (wmk-1/wmk-2 and wmk-3/wmk-4). Of these, wmk-1 or wmk-3 killed almost all Drosophila melanogaster individuals when transgenically overexpressed. Dual expression of wmk-3 and wmk-4 killed all males and rescued females. We propose a novel hypothesis wherein horizontally transmitted proto-Wolbachia with a single wmk killed both sexes, and tandem duplication of wmk allowed an evolutionary transition to a vertically transmitted symbiont, causing male-killing. Our study highlights the bacteriophage as a critical driver of the evolution of male-killing and argues for a conserved male-killing mechanism in diverse insects.

Bumblebees are important pollinators of wild and agricultural plants but recently have been declining due to various stressors, such as pesticides and diseases. Because of the haplo-diploid sex determination system in hymenopterans, experiments using micro-colonies (small sub colonies without a queen) to identify risks to bumblebee health are limited as they are only able to produce males. Therefore, an experimental protocol for rearing bumblebee larvae in vitro is needed to better understand effects on worker larvae. Here, we aimed to establish a rearing method for larvae of Bombus terrestris for use in risk assessment assays. To confirm the validity of our rearing method, we tested two insecticides used for tomato cultivation, chlorfenapyr and dinotefuran. Bombus terrestris larvae fed with a high nutrient quantity and quality diet increased growth per day. All chlorfenapyr-exposed individuals died within 10 days at 2000-fold dilution, an application dose used for tomatoes. There were significant differences in adult emergence rate among almost all chlorfenapyr treatments. On the other hand, sublethal dinotefuran-exposure did not affect rates of pupation and adult emergence, growth, or larval and pupal periods. Although larvae were smaller than in the natural colony, this rearing method for B. terrestris larvae proved to be effective at evaluating realistic sub-colonies to pesticide exposures.

Male killing, the phenomenon of male death during development, is considered to be one of the advantageous strategies exerted by maternally transmitted microbes. Male killing has attracted interest in the fields of evolutionary biology and ecology for decades; however, little is known about its mechanism and origin. Here, we characterized and compared the effects of three distinct male killers, Wolbachia (Alphaproteobacteria), Spiroplasma (Mollicutes), and Osugoroshi virus (OGV) (Partitiviridae) in the tea pest moth Homona magnanima (Lepidoptera, Tortricidae). Regardless of the genetic sex (male: ZZ; female: ZW), female specific splice variants of the doublesex gene (dsx), a downstream regulator of the sex-determining gene cascade, was expressed in H. magnanima harbored either male-killing Wolbachia or Spiroplasma. However, OGV and non-male-killing Wolbachia did not alter dsx splicing. RNA sequencing and quantitative PCR assays demonstrated that male-killing Wolbachia impaired the host’s dosage compensation system by altering the global gene expression of the Z chromosome (corresponding to Bombyx mori chromosome 1 and 15) in males, whereas Spiroplasma did not affect dosage compensation. In contrast, the partiti-like virus OGVs did not affect sex-determination cascades or dosage compensation systems. Besides, male killers distinctly altered host gene expression and metabolomes associated with physiology, morphology, and diverse metabolic pathways. Moreover, Wolbachia and Spiroplasma infections triggered abnormal apoptosis only in male embryos. These findings suggest that distantly related microbes employ distinct machineries to kill identical host males, which have been acquired through independent evolutionary processes.ImportanceMale-killing caused by diverse microbes has attracted substantial attention. However, it remains unclear how such male killers have evolved similar phenotypes, in part because male-killing mechanisms have been studied using different insect models. Here, by comparing three phylogenetically distinct male killers, Wolbachia, Spiroplasma, and a partiti-like virus, in an identical host, we provide evidence that microbes can affect male viability through distinct machinery, demonstrating distinct evolutionary scenarios for microbes to acquire make-killing ability. These findings provide insight into new directions for studying microbe–host interactions.

Funding Acknowledgements Type of funding sources: None. Background and Aims n-3 polyunsaturated fatty acids (PUFAs) have an inhibitory effect against the development of coronary artery disease (CAD). However, whether n-6 PUFAs, dihomo-gamma-linolenic acid (DGLA), and arachidonic acid (AA) play a role in the development of CAD remains unclear. This study investigated the association between PUFAs and the risk of developing acute coronary syndrome (ACS) using the lipid and PUFAs data of patients who received percutaneous coronary intervention (PCI) for either non-emergent conditions (staged group) or ACS (ACS group). Methods We retrospectively evaluated 433 patients who underwent PCI between 2014 and 2021. The patients were divided into the ACS group (n = 18) and the staged group (n = 132). The lipid and PUFAs values of each patient between the two groups were compared. Moreover, to investigate the correlation between n-6 PUFAs levels and ACS, the effects of confounding factors such as use of strong statins and low-density cholesterol (LDL-C) levels were adjusted. Results The ACS group had higher n-6 PUFAs levels than the staged group (DGLA: 36.8 ug/mL vs. 29.6 ug/mL; AA: 203.3 ug/mL vs. 145.8 ug/mL). Furthermore, the analysis of covariance adjusted for LDL-C levels showed a significant difference between the two groups in terms of DGLA and AA levels. Conclusions The n-3 PUFAs levels did not significantly differ between the staged and ACS groups. Moreover, the ACS group had higher DGLA and AA levels and lower n-3 PUFAs/AA ratios than the staged group. Therefore, excess n-6 PUFAs may be a risk factor for ACS.

Tortricidae (Lepidoptera), commonly known as tortrix or leafroller moths, comprises many agricultural and forestry pests, which cause serious agricultural losses. To understand the biology of such pest moths, fundamental techniques have been in high demand. Here, methods for mass-rearing, observations, and molecular studies are developed using two tea tortrix, Homona magnanima and Adoxophyes honmai (Lepidoptera: Tortricidae). Insects were mass-reared with sliced artificial diet and maintained by inbreeding for over 100 generations by considering their biological characteristics. Insects have various sex dimorphisms; hence it is difficult to distinguish the sex during the developing stages, which have prevented subsequent assays. The present work highlighted that the sex of tortricids larvae could be determined by observing testes or lactic-acetic orcein staining to visualize the female-specific W chromosome. Moreover, using the sex determination methods, the present study enabled nucleic acid extractions from sex determined embryos and application toward high throughput sequencing. These tips are applicable for other pest insects and will facilitate further morphological and genetic studies.

Microsporidia are spore-forming intracellular parasites of various invertebrates and vertebrates. Vairimorpha bombi negatively affects the fitness of bumblebees and its prevalence correlates with declining bumblebee populations. The invasive alien species Bombus terrestris colonized Japan and possibly introduced new parasites. To assess the infection prevalence of V. bombi in Japanese bumblebees and B. terrestris, we investigated V. bombi infections using PCR and microscopy. The prevalence of sporulating V. bombi infections in three Bombus s. str. species/subspecies was low, whereas that of non/low-sporulating Vairimorpha sp. infections in three Diversobombus species/subspecies was high. Invasive B. terrestris showed low prevalence of non/low-sporulating V. bombi infections and shared the same V. bombi haplotype with B. hypocrita found in Hokkaido, where B. terrestris is present, and in Honshu, where B. terrestris is absent. Although V. bombi may have been introduced with B. terrestris colonies imported from Europe, it seems to be originally distributed in Japan. Furthermore, a new Vairimorpha sp. was found in Japanese bumblebee species. V. bombi and Vairimorpha sp. showed different organ and host specificities in bumblebees. There are no reports on the specific effects of other Vairimorpha spp. on bumblebees; further studies are needed to clarify the individual characteristics of Vairimorpha spp.

Coconut rhinoceros beetle (CRB), Oryctes rhinoceros, is a pest of palm trees in the Pacific. Recently, a remarkable degree of palm damage reported in Guam, Hawaii, Papua New Guinea and Solomon Islands has been associated with a particular haplotype (clade I), known as “CRB-G”. In the Palau Archipelago, both CRB-G and another haplotype (clade IV) belonging to the CRB-S cluster coexist in the field. In this study, more than 75% of pheromone trap-captured adults of both haplotypes were Oryctes rhinoceros nudivirus (OrNV)-positive by PCR. No significant difference in OrNV prevalence between the haplotypes was detected. In PCR-positive CRB-G tissue specimens from Palau, viral particles were observed by electron microscopy. Hemocoel injection of CRB larvae with crude virus homogenates from these tissues resulted in viral infection and mortality. OrNV isolated from Palauan-sourced CRB was designated as OrNV-Palau1. Both OrNV-Palau1 and OrNV-X2B, a CRB biological control isolate released in the Pacific, were propagated using the FRI-AnCu-35 cell line for production of inoculum. However, the OrNV-Palau1 isolate exhibited lower viral production levels and longer larval survival times compared to OrNV-X2B in O. rhinoceros larvae. Full genome sequences of the OrNV-Palau1 and -X2B isolates were determined and found to be closely related to each other. Altogether these results suggest CRB adults in Palau are infected with a less virulent virus, which may affect the nature and extent of OrNV-induced pathology in Palauan populations of CRB.

For regulatory purposes, the name ‘Asian gypsy moth’ refers to a group of closely related Asian Lymantria species and subspecies whose female moths display flight capability, a trait believed to confer enhanced invasiveness relative to the European gypsy moth, Lymantria dispar dispar, whose females are flightless. Lymantria albescens and Lymantria postalba are Asian gypsy moths occurring in the southern Ryukyu Islands and in the northern Ryukyu and adjacent Kyushu and Shikoku Islands of Japan, respectively. Although once considered subspecies of L. dispar, their status as distinct species, relative to the latter, is now well established. While postalba was subsequently considered a subspecies of L. albescens, largely on the basis of differences in forewing ground colour in males, both taxa were later given distinct species status by Pogue & Schaefer (2007) following their revision of the genus Lymantria. Here, we re‐examined the validity of this revised status through the sequencing of a large portion of the mitochondrial genome (c. 60%) and multiple nuclear marker genes [elongation factor 1‐alpha (Ef‐1α), wingless (Wgl), internal transcribed spacer 2 (ITS‐2), ribosomal protein S5 (RpS5)] in representative specimens of both taxa and other Lymantria species, including L. monacha, L. xylina, L. mathura and members of the L. dispar + L. umbrosa clade. A comparison of the number of substitutions in these genomic regions among the taxa we considered showed lower or equivalent variation between L. albescens and L. postalba compared with subspecies of L. dispar, for mitochondrial and nuclear sequences, respectively. This finding was reflected in the maximum likelihood trees generated independently for mitochondrial and nuclear data, where L. albescens and L. postalba formed, in both analyses, a short‐branch sister clade basal to the L. dispar + L. umbrosa clade. We further sequenced three markers [cytochrome c oxydase 1 (COI), EF‐1α, Wgl] in multiple L. albescens–L. postalba specimens collected along a south‐to‐north transect across the Ryukyu Arc and observed no clear distinction among the sampled specimens as a function of taxonomic designation. We conclude that L. albescens and L. postalba form a single species, with postalba representing a darker‐winged morph along an apparent south‐to‐north wing colour cline. Accordingly, L. postalba is relegated to synonymy under L. albescens (syn.n.).

Wolbachia are inherited intracellular bacteria that cause male-specific death in some arthropods, called male-killing. To date, three Wolbachia strains have been identified in the oriental tea tortrix Homona magnanima (Tortricidae, Lepidoptera); however, none of these caused male-killing in the Japanese population. Here, we describe a male-killing Wolbachia strain in Taiwanese H. magnanima. From field-collected H. magnanima, two female-biased host lines were established, and antibiotic treatments revealed Wolbachia (wHm-t) as the causative agent of male-killing. The wsp and MLST genes in wHm-t are identical to corresponding genes in the nonmale-killing strain wHm-c from the Japanese population, implying a close relationship of the two strains. Crossing the Japanese and Taiwanese H. magnanima revealed that Wolbachia genotype rather than the host genetic background was responsible for the presence of the male-killing phenotype. Quantitative PCR analyses revealed that the density of wHm-t was higher than that of other Wolbachia strains in H. magnanima, including wHm-c. The densities of wHm-t were also heterogeneous between host lines. Notably, wHm-t in the low-density and high-density lines carried identical wsp and MLST genes but had distinct lethal patterns. Furthermore, over 90% of field-collected lines of H. magnanima in Taiwan were infected with wHm-t, although not all host lines harboring wHm-t showed male-killing. The host lines that showed male-killing harbored a high density of Wolbachia compared to the host lines that did not show male-killing. Thus, the differences in the phenotypes appear to be dependent on biological and genetic characteristics of closely related Wolbachia strains.

The gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae), is distributed throughout most of the northern hemisphere and known as one of the most significant insect pest both in its native and introduced regions. The aim of this study was to examine whether there are differences in the outbreak pattern of L. dispar, and these differences can be explained by different subspecies or divided populations across Japan for appropriate management of L. dispar. We reviewed the records of outbreaks of L. dispar in Japan using available literature and internet sources. We also examined phylogenetic relationships between L. dispar populations in Japan and other global populations to clarify current debated classification of the Japanese L. dispar. We then estimated the distribution of Japanese L. dispar with a species distribution model (Maxent). Phylogenetic mtDNA analysis revealed that Lymantria species consisted of four clusters: the first cluster contained three L. dispar subspecies from Europe and continental Asia (classified into L. dispar dispar), the populations of Honshu and western Hokkaido (L. dispar asiatica), and the central and eastern Hokkaido populations of L. dispar hokkaidoensis, while the second and third clusters comprised L. albescens and L. postalba, respectively. According to historical records, L. dispar outbreaks occur in approximately 11 year-cycles in Hokkaido, regardless of its classification at the subspecies level. Within northeastern Japan, L. dispar outbreaks occurred simultaneously from 2014 to 2015, whereas there were few reports of outbreaks in southwestern Japan. The distribution ranges of L. dispar were determined mainly by the maximum depth of snow cover (100–250 cm), forest patch area (0–0.6 km2), and elevation (600–1400 m); clearly divided at the species level but not at the subspecies or divided population level. On the other hand, the forest types are likely to contribute to the population dynamics and geographical distribution of L. dispar.

Study question Do patients with inherited thrombophilia associated to RIF and RPL benefit from anticoagulant therapy? Summary answer Low molecular weight heparin (LMWH) in patients with medium and high risk of hereditary thrombophilia, associated with RIF could improve the reproductive prognosis. What is known already Thrombophilia is a condition that can be acquired and/or inherited genetically, that is characterized by the predisposition of patients to form venous and arterial thromboembolic events. Inherited thrombophilia has been associated with different complications during pregnancy, such as RPL. Genetic variants linked to hereditary thrombophilia can be classified by the thromboembolic risk: low (F12, F13A1, FGB), medium (MTHFR, PROCR, PROS1, SERPINC1, SERPINC1 PAI–1) and high (F2, F5, GP1BA), according to Martinez - Zamora. RPL rate may reduce with anticoagulant therapy. However, there is no conclusive evidence that prophylactic treatment improves the pregnancy rate in infertile women during IVF. Study design, size, duration We performed a prospective observational study which included patients referred to Ceras Clinic between March 2018 and March 2020, due to RPL (n = 38) and RIF (n = 40). All patients underwent genetic analysis for hereditary thrombophilia(F13, F2, F5, FGB, GP1BA, MTHFR C677T, MTHFR A1298C, PAI1,, PROCR, SERPIN1 CM910058, SERPIN 1 CM920113 ,F12, PROS1) by Sanger sequencing. The characteristics of anticoagulant therapy with clinical pregnancy rate and LBR were analyzed, using chi-squared test with STATA version 16. Participants/materials, setting, methods Patients have been included in the study according to their past medical history (stroke or myocardial infarction, personal or familiar history of deep vein thrombosis or pulmonary embolism, smoking, hormone replacement therapy), and reproductive history. Two groups were formed, the first group (n = 40) corresponds to RIF, and the second (n = 38), RPL. Genetic study of hereditary thrombophilia (11 genes) was performed to examine the genetic risk and assess the administration of anticoagulant therapy. Main results and the role of chance The prevalence of pathological antecedents in patients with RIF and RPL was not statistically significant (p > 0,05), indicating that the factors that contribute to poor reproductive outcomes in these two groups of patients could be similar. Patients with RIF had a medium risk of thrombophilia in 65%, followed by low risk in 32.5% and high risk in 2.5%. RPL group presented 78.95%, 15.79% and 5.26%, respectively. All patients with medium and high risk for trombophilia received anticoagulation. Clinical pregnancy rate (69.7%) and live birth rate (63.64%) were not statistically significant (p > 0.05) in RPL with anticoagulant therapy, compared to RPL group who did not received treatment (clinical pregnancy rate and live birth rate in 60%). Therefore, it is proposed that there may be other factors associated with abortions that require investigation. However, clinical pregnancy rate (77.14%) and live birth rate (74.29%) were statistically significant (p Limitations, reasons for caution The small number of patients assessed is the main limitation of this work. Larger studies must be designed to accurately determine participation of each mutation associated with recurrent implantation failure and recurrent pregnancy loss. The role of anticoagulant therapy should be evaluated in randomized clinical trials. Wider implications of the findings: Establishing a stronger evidence base implies that future studies should include large population groups. It is primordial to assess whether it is cost-effective to determine the risk of inherited thrombophilia in RIL and RPL, to increase the live birth rate by anticoagulant therapy. The information is controversial to this day. Trial registration number ‘not applicable’

The gypsy moth, Lymantria dispar, is a notorious forest defoliator, and various pathogens are known to act as natural regulators of its population density. As a widespread herbivore with a broad range of inhabited areas and host plants, it is potentially exposed to parasitic microorganisms from other insect hosts. In the present paper, we determined the susceptibility of gypsy moth larvae to the microsporidium Nosema pyrausta from the European corn borer, Ostrinia nubilalis. Gypsy moth samples from two localities of Western Siberia were used. N. pyrausta developed infections in the salivary gland and adipose tissue of gypsy moth prepupae and pupae, forming spore masses after 30 days of alimentary exposure to the second instar larvae. Among the experimental groups, the infection levels ranged from 0 to 9.5%. Effects of a covert baculovirus infection, phenylthiourea pretreatment and feeding insects on an artificial diet versus natural foliage were not significant in terms of microsporidia prevalence levels. Thus, L. dispar showed a low level of susceptibility to a non-specific microsporidium. It can be referred to as a resistant model host and not an appropriate substitute host for laboratory propagation of the microsporidium.

We present the first observation by the Telescope Array Surface Detector (TASD) of the effect of thunderstorms on the development of cosmic ray single count rate intensity over a 700 km$^{2}$ area. Observations of variations in the secondary low-energy cosmic ray counting rate, using the TASD, allow us to study the electric field inside thunderstorms, on a large scale, as it progresses on top of the 700 km$^{2}$ detector, without dealing with the limitation of narrow exposure in time and space using balloons and aircraft detectors. In this work, variations in the cosmic ray intensity (single count rate) using the TASD, were studied and found to be on average at the $\sim(0.5-1)\%$ and up to 2\% level. These observations were found to be both in excess and in deficit. They were also found to be correlated with lightning in addition to thunderstorms. These variations lasted for tens of minutes; their footprint on the ground ranged from 6 to 24 km in diameter and moved in the same direction as the thunderstorm. With the use of simple electric field models inside the cloud and between cloud to ground, the observed variations in the cosmic ray single count rate were recreated using CORSIKA simulations. Depending on the electric field model used and the direction of the electric field in that model, the electric field magnitude that reproduces the observed low-energy cosmic ray single count rate variations was found to be approximately between 0.2-0.4 GV. This in turn allows us to get a reasonable insight on the electric field and its effect on cosmic ray air showers inside thunderstorms.

In this paper we report the first close, high-resolution observations of downward-directed terrestrial gamma-ray flashes (TGFs) detected by the large-area Telescope Array cosmic ray observatory, obtained in conjunction with broadband VHF interferometer and fast electric field change measurements of the parent discharge. The results show that the TGFs occur during strong initial breakdown pulses (IBPs) in the first few milliseconds of negative cloud-to-ground and low-altitude intracloud flashes, and that the IBPs are produced by a newly-identified streamer-based discharge process called fast negative breakdown. The observations indicate the relativistic runaway electron avalanches (RREAs) responsible for producing the TGFs are initiated by embedded spark-like transient conducting events (TCEs) within the fast streamer system, and potentially also by individual fast streamers themselves. The TCEs are inferred to be the cause of impulsive sub-pulses that are characteristic features of classic IBP sferics. Additional development of the avalanches would be facilitated by the enhanced electric field ahead of the advancing front of the fast negative breakdown. In addition to showing the nature of IBPs and their enigmatic sub-pulses, the observations also provide a possible explanation for the unsolved question of how the streamer to leader transition occurs during the initial negative breakdown, namely as a result of strong currents flowing in the final stage of successive IBPs, extending backward through both the IBP itself and the negative streamer breakdown preceding the IBP., Comment: Typo fixed and reference added. Manuscript is 36 pages. Supplemental Information is 42 pages. This paper is to be published in the Journal of Geophysical Research: Atmospheres. Online data repository: Open Science Framework DOI: 10.17605/OSF.IO/Z3XDA

Late male-killing, a male-specific death after hatching, is a unique phenotype found in Homona magnanima, oriental tea tortrix. The male-killing agent was suspected to be an RNA virus, but details were unknown. We herein successfully isolated and identified the putative male-killing virus as Osugoroshi viruses (OGVs). The three RNA-dependent RNA polymerase genes detected were phylogenetically related to Partitiviridae, a group of segmented double-stranded RNA viruses. Purified dsRNA from a late male-killing strain of H. magnanima revealed 24 segments, in addition to the RdRps, with consensus terminal sequences. These segments included the previously found male-killing agents MK1068 (herein OGV-related RNA16) and MK1241 (OGV-related RNA7) RNAs. Ultramicroscopic observation of purified virions, which induced late male-killing in the progeny of injected moths, showed sizes typical of Partitiviridae. Mathematical modeling showed the importance of late male-killing in facilitating horizontal transmission of OGVs in an H. magnanima population. This study is the first report on the isolation of partiti-like virus from insects, and one thought to be associated with late male-killing, although the viral genomic contents and combinations in each virus are still unknown.

Background Anti-Gerbich (Ge) alloantibody against high-frequency erythrocyte antigen is extremely rare. Owing to incomplete evidence regarding the degree and severity of adverse events induced by hemolytic transfusion reactions, the transfusion management often remains cumbersome in these patients. We report an anti-Ge alloantibody positive patient with hepatocellular carcinoma (HCC) who underwent central hepatectomy (CH) without the need for an allogeneic blood transfusion. Case presentation A 76-year-old Japanese woman was diagnosed with HCC measuring 9.5 × 8.0 cm in segments 4, 5, and 8 of the liver. This patient with anti-Ge alloantibody had a history of two pregnancies without transfusion. CH was planned, and based on the suggestion from the multidisciplinary team meeting, preoperative autologous donation (PAD) and acute normovolemic hemodilution (ANH) were performed. CH was successfully performed by using CUSA and Thunderbeat® with Pringle maneuver and infra-hepatic inferior vena cava clamping without perioperative need for an allogeneic blood transfusion. She has been alive without recurrence after a follow-up period of 45 months. Conclusion To our knowledge, this is the first case report of hepatectomy in a patient with anti-Ge alloantibody. A multidisciplinary team approach, PAD and ANH, and bloodless liver surgical techniques appear to be useful for major hepatectomy in patients with extremely rare blood type.

Cyclosporine A is known to be effective in some genetic podocyte injury. However, the efficacy of cyclosporine A depends on the degree of histopathological findings, and the relationship between long-term use and renal prognosis remains unknown. Frasier syndrome is a rare genetic disorder caused by intronic mutations in WT1, and is characterized by progressive glomerulopathy, a 46,XY disorder of sex development, and an increased risk of gonadoblastoma. We report here a 16-year-old phenotypically female patient with Frasier syndrome. A renal biopsy at the age of seven years showed segmentally effaced podocyte foot processes with no evidence of glomerulosclerosis. Steroid-resistant proteinuria progressed to the nephrotic range at the age of 10 years, which responded to once-daily administration of cyclosporine A with low two-hour post-dose cyclosporine A (C2) levels; she then achieved stable partial remission in combination with renin-angiotensin system (RAS) blockade. At the age of 12 years, examinations for delayed puberty confirmed the diagnosis of Frasier syndrome. The second renal biopsy showed widespread foot process effacement and a minor lesion of segmental glomerulosclerosis without findings suggestive of cyclosporine A nephropathy. She maintained partial remission and normal renal function with the continuation of once-daily low-dose cyclosporine A. The C2 levels required for the remission were between 212 and 520 ng/ml. Cyclosporine A dosages sufficient for maintaining the C2 levels were 1.1-1.2 mg/kg per day. In conclusion, the long-lasting treatment of once-daily low-dose cyclosporine A with RAS inhibition was effective for induction and maintenance of partial remission in Frasier syndrome.

Central biliary metastasis from colorectal cancer (CRC) is a rare manifestation. Given the rarity of the lesion, cases with central biliary metastasis from CRC pose challenges in identifying the proper treatment strategy. We present a case of stage IV CRC with synchronous multiple liver metastases and central biliary invasion. We also conducted a literature search and reviewed the reports on central biliary metastasis of CRC treated by major hepatectomy combined with extrahepatic bile duct resection. The patient underwent colon resection, partial hepatectomy, and transileocecal portal embolization (TIPE) as the first procedure, which was followed by an extended right hepatectomy and extrahepatic bile duct resection. Adjuvant chemotherapy was introduced, and the patient was doing well with no recurrence at 17 months after the first procedure. A liberal radical and aggressive resection in a multimodal treatment strategy could offer a favorable outcome in patients with a rare central biliary manifestation of CRC.

Bursts of gamma ray showers have been observed in coincidence with downward propagating negative leaders in lightning flashes by the Telescope Array Surface Detector (TASD). The TASD is a 700~square kilometer cosmic ray observatory located in southwestern Utah, U.S.A. In data collected between 2014 and 2016, correlated observations showing the structure and temporal development of three shower-producing flashes were obtained with a 3D lightning mapping array, and electric field change measurements were obtained for an additional seven flashes, in both cases co-located with the TASD. National Lightning Detection Network (NLDN) information was also used throughout. The showers arrived in a sequence of 2--5 short-duration ($\le$10~$\mu$s) bursts over time intervals of several hundred microseconds, and originated at an altitude of $\simeq$3--5 kilometers above ground level during the first 1--2 ms of downward negative leader breakdown at the beginning of cloud-to-ground lightning flashes. The shower footprints, associated waveforms and the effect of atmospheric propagation indicate that the showers consist primarily of downward-beamed gamma radiation. This has been supported by GEANT simulation studies, which indicate primary source fluxes of $\simeq$$10^{12}$--$10^{14}$ photons for $16^{\circ}$ half-angle beams. We conclude that the showers are terrestrial gamma-ray flashes (TGFs), similar to those observed by satellites, but that the ground-based observations are more representative of the temporal source activity and are also more sensitive than satellite observations, which detect only the most powerful TGFs.

Endosymbiotic bacterium Wolbachia interacts with host in either a mutualistic or parasitic manner. Wolbachia is frequently identified in various arthropod species, and to date, Wolbachia infections have been detected in different insects. Here, we found a triple Wolbachia infection in Homona magnanima, a serious tea pest, and investigated the effects of three infecting Wolbachia strains (wHm-a, -b, and -c) on the host. Starting with the triple-infected host line (Wabc), which was collected in western Tokyo in 1999 and maintained in laboratory, we established an uninfected line (W−) and three singly infected lines (Wa, Wb, and Wc) using antibiotics. Mating experiments with the host lines revealed that only wHm-b induced cytoplasmic incompatibility (CI) in H. magnanima, with the intensities of CI different between the Wb and Wabc lines. Regarding mutualistic effects, wHm-c shortened larval development time and increased pupal weight in both the Wc and Wabc lines to the same extent, whereas no distinct phenotype was observed in lines singly infected with wHm-a. Based on quantitative PCR analysis, Wolbachia density in the Wa line was higher than in the other host lines (p

A granulovirus (GV) that produces occlusion bodies (OBs) having an unusual morphology was found in an Adoxophyes sp. (Lepidoptera: Tortricidae) larva in a tea field in Miyazaki Prefecture, Japan. This isolate is considered to be a mutant of Adoxophyes orana granulovirus, designated AdorGV-M, because the nucleotide sequence of its genome is 99.7% identical to that of an English isolate of AdorGV, AdorGV-E. AdorGV-E produces typical ovocylindrical OBs that contain one occlusion-derived virus (ODV) per OB. On the other hand, AdorGV-M produces large cuboidal OBs, but the number of ODVs per OB was unknown. In this study, we quantified viral DNA in OBs of both AdorGV-E and -M, and determined the number of ODVs occluded in an OB of AdorGV-M. The two isolates had the same quantity of viral DNA in each OB, and we thus confirmed that one OB of AdorGV-M contains one ODV. To investigate the process of OB formation, fat body tissue of A. honmai larvae inoculated with each isolate was observed in a time course by transmission electron microscopy, and OB sizes were measured from micrographs. The main difference in OB formation was that AdorGV-M required more time to mature than AdorGV-E. In AdorGV-E, ODVs began to be covered from one end with an ovocylindrical OB at 96 h post-inoculation (hpi), and most of them were completely occluded at 120 hpi. Occlusion of AdorGV-M ODVs also began at 96 hpi, but the OB shape was cuboidal. Moreover, the OB size of AdorGV-M was similar to that of AdorGV-E at 120 hpi, but continued to grow until 192 hpi. AdorGV-M thus took more time to complete OB formation. Consequently, AdorGV-E has mature OBs with a diameter 0.22 μm and length 0.39 μm, but those of AdorGV-M are 1.34 × 1.23 μm.

A method for increasing compositional or material contrast of a standard semiconductor BSE detector in a scanning electron microscope (SEM) by compensation of the topographic contrast component is proposed. Compensation is based on the physical properties of backscattered electron emission and topography information of the specimen's surface. Three analytical and semi-empirical compensation algorithms employing different physical models and approximations are implemented and compared to conventional BSE signals to show the effectivity of the proposed compensation approach.

The Telescope Array (TA) experiment is designed to detect air showers induced by ultra high energy cosmic rays. The TA ground Surface particle Detector (TASD) observed several short-time bursts of air shower like events. These bursts are not likely due to chance coincidence between single shower events. The expectation of chance coincidence is less than 10 − 4 for five-year's observation. We checked the correlation between these bursts of events and lightning data, and found evidence for correlations in timing and position. Some features of the burst events are similar to those of a normal cosmic ray air shower, and some are not. On this paper, we report the observed bursts of air shower like events and their correlation with lightning.

Motivated by the detection of a significant dipole structure in the arrival directions of ultrahigh-energy cosmic rays above 8 EeV reported by the Pierre Auger Observatory (Auger), we search for a large-scale anisotropy using data collected with the surface detector array of the Telescope Array Experiment (TA). With 11 years of TA data, a dipole structure in a projection of the right ascension is fitted with an amplitude of 3.3+- 1.9% and a phase of 131 +- 33 degrees. The corresponding 99% confidence-level upper limit on the amplitude is 7.3%. At the current level of statistics, the fitted result is compatible with both an isotropic distribution and the dipole structure reported by Auger., Comment: 6 pages, 3 figures, 1 table, Proofed title. Added journal reference and DOI