Subsequent Replication Research Articles

On the influence of fuzzy perception of the time passage speed on the solutions of optimization planning problems

The paper considers an approach to the construction of fuzzy structured numerical sets based on the formation of a fuzzy original of a special type with subsequent replication on the number axis. The constructive principle of fuzzy original formalization is to define a fuzzy triangular number with the appropriate support. A variant of the formation of fuzzy numerical sets, formalizing the "fast" and "slow" passage of time, is proposed. The developed technique allows us to propose a solution to the problem associated with the formalization of the subjective perception of the time reference in processes involving a person, to obtain results and evaluate the impact of fuzzy description and taking into account the dynamics of the time reference on the solution of various optimization problems. A general approach to the formation and solution of fuzzy linear optimization problems is considered, the transition from fuzzy to parametric formulation in the Bellman-Zade form is described. Examples of the use of fuzzy flow of time for different statements of tasks that arise when determining the order of the set of tasks within a given time interval with or without additional restrictions on the execution process are considered. An approach is proposed for the correction of the initial time distribution plans, taking into account different rates of time counting. A mathematical model of the fuzzy traveling salesman problem is formulated as a problem of finding a route to visit a given number of cities without repetitions with a minimum travel time with time parameters specified as right fuzzy numbers, the support value in which depends on various external conditions and factors. The results of calculations of solutions of fuzzy optimization problems are obtained. The influence of the speed of the passage of time on the solution of the considered optimization problems is determined.

DNA primase large subunit is an essential plant gene for geminiviruses, putatively priming viral ss-DNA replication.

The family of Geminiviridae consists of more than 500 circular single-stranded (ss) DNA viral species that can infect numerous dicot and monocot plants. Geminiviruses replicate their genome in the nucleus of a plant cell, taking advantage of the host's DNA replication machinery. For converting their DNA into double-stranded DNA, and subsequent replication, these viruses rely on host DNA polymerases. However, the priming of the very first step of this process, i.e. the conversion of incoming circular ssDNA into a dsDNA molecule, has remained elusive for almost 30 years. In this study, sequencing of melon (Cucumis melo) accession K18 carrying the Tomato leaf curl New Delhi virus (ToLCNDV) recessive resistance quantitative trait locus (QTL) in chromosome 11, and analyses of DNA sequence data from 100 melon genomes, showed a conservation of a shared mutation in the DNA Primase Large subunit (PRiL) of all accessions that exhibited resistance upon a challenge with ToLCNDV. Silencing of (native) Nicotiana benthamiana PriL and subsequent challenging with three different geminiviruses showed a severe reduction in titers of all three viruses, altogether emphasizing an important role of PRiL in geminiviral replication. A model is presented explaining the role of PriL during initiation of geminiviral DNA replication, i.e. as a regulatory subunit of primase that generates an RNA primer at the onset of DNA replication in analogy to DNA Primase-mediated initiation of DNA replication in all living organisms.

Physicochemical properties of green synthesised ZnO nanoparticles and utilisation for treatment of breast cancer

Metal oxide nanoparticles with unique properties have recently attracted much attention as a potential smart material for treatment of cancer. The inherent structural and ionic defects in ZnO nanoparticles allow the formation of intracellular superoxide and hydroxyl radicals after entering a cancer cell causing damage on DNA and preventing subsequent replication. However, the full extent of the DNA damage and the knock-on effect with respect to protein synthesis is not copiously comprehended. This study tries to tune the cytotoxic activity of ZnO nanoparticles in virtue of physicochemical property against human breast cancer cell (MCF-7) via reactive oxygen species (ROS) induced DNA damage leading to apoptosis. In order to produce an array of different size and shape of ZnO nanoparticles with varying structural defects, this study utilised the unique phytochemicals of Terminalia arjuna, Swertia chirayita, and Psidium guajava as the active reducing agents for the formation of the nanoparticle from the mineral salt. Variations in the created nanoparticle size and relative structural defects have caused a consequential surge in ROS production inside cancer cells resulting in enhanced effectiveness of ZnO nanoparticles for cancer treatment.

Therapeutic Potential of PTBP1 Inhibition, If Any, Is Not Attributed to Glia-to-Neuron Conversion.

A holy grail of regenerative medicine is to replenish the cells that are lost due to disease. The adult mammalian central nervous system (CNS) has, however, largely lost such a regenerative ability. An emerging strategy for the generation of new neurons is through glia-to-neuron (GtN) conversion in vivo, mainly accomplished by the regulation of fate-determining factors. When inhibited, PTBP1, a factor involved in RNA biology, was reported to induce rapid and efficient GtN conversion in multiple regions of the adult CNS. Remarkably, PTBP1 inhibition was also claimed to greatly improve behaviors of mice with neurological diseases or aging. These phenomenal claims, if confirmed, would constitute a significant advancement in regenerative medicine. Unfortunately, neither GtN conversion nor therapeutic potential via PTBP1 inhibition was validated by the results of multiple subsequent replication studies with stringent methods. Here we review these controversial studies and conclude with recommendations for examining GtN conversion in vivo and future investigations of PTBP1.

Chelerythrine inhibits HSV-1 infection by suppressing virus binding to the cells

ObjectiveHerpes simplex virus type 1 (HSV-1) is a worldwide pathogen and mainly puts immunocompromised patients at higher risk of various complications. This research aims to demonstrate the antiviral activity of chelerythrine (CHE), an alkaloid compound derived from the traditional Chinese medicine Mcleaya cordata, towards the HSV-1 F strain. MethodsBioassay-guided fractionation was carried out in order to isolate the antiviral compound CHE. The plaque reduction assay was used to evaluate the antiviral activity of CHE. The combination of quantitative PCR (qPCR) and western blot was used to elucidate whether CHE interferes with viral DNA synthesis, followed by time-of-addition assay to examine which steps of HSV-1 infection CHE affected. The immunofluorescent assay and transmission electron microscopy assay were performed to further analyze the intracellular localization and the ultrastructural features of HSV-1 virions with the treatment of CHE. The binding affinity in CHE to targeted viral glycoprotein was investigated by surface plasmon resonance (SPR) assay and docking simulation. ResultsCHE displayed a significant antiviral effect against HSV-1 at 0.312 to 2.5 µM with a viral titer reduction of 29.38% to 61.02%. CHE had an EC50 value of 1.78 µM against HSV-1, and the selectivity index (SI) for CHE was 4.07. Additionally, CHE inhibits ICP0 expression in the immediate early stage of infection, which further affected the early stage of infection, and subsequent viral replication. Results of the SPR assay showed that CHE had strong-binding affinity to gB with a KD value of 7.80 × 107 M, while gD with a value of 1.18 × 106 M. TEM revealed that CHE can clearly inhibit the virion release. In silico experiments further confirmed that CHE had low binding free energy in the activity sites of gB and gD. ConclusionThis study revealed that CHE directly targeted viral gB or gD as a potent glycoprotein inhibitor, which suggested that CHE could be a virus entry inhibitor for HSV-1 treatment.

Direct Processing of PVD Hard Coatings via Focused Ion Beam Milling for Microinjection Molding

Hard coatings can be applied onto microstructured molds to influence wear, form filling and demolding behaviors in microinjection molding. As an alternative to this conventional manufacturing procedure, "direct processing" of physical-vapor-deposited (PVD) hard coatings was investigated in this study, by fabricating submicron features directly into the coatings for a subsequent replication via molding. Different diamondlike carbon (DLC) and chromium nitride (CrN) PVD coatings were investigated regarding their suitability for focused ion beam (FIB) milling and microinjection molding using microscope imaging and areal roughness measurements. Each coating type was deposited onto high-gloss polished mold inserts. A specific test pattern containing different submicron features was then FIB-milled into the coatings using varied FIB parameters. The milling results were found to be influenced by the coating morphology and grain microstructure. Using injection-compression molding, the submicron structures were molded onto polycarbonate (PC) and cyclic olefin polymer (COP). The molding results revealed contrasting molding performances for the studied coatings and polymers. For CrN and PC, a sufficient replication fidelity based on AFM measurements was achieved. In contrast, only an insufficient molding result could be obtained for the DLC. No abrasive wear or coating delamination could be found after molding.

Spatiotemporal view of malignant histogenesis and macroevolution via formation of polyploid giant cancer cells

To understand how malignant tumors develop, we tracked cell membrane, nuclear membrane, spindle, and cell cycle dynamics in polyploid giant cancer cells (PGCCs) during the formation of high-grade serous carcinoma organoids using long-term time-lapse imaging. Single cells underwent traditional mitosis to generate tissue with uniform nuclear size, while others formed PGCCs via asymmetric mitosis, endoreplication, multipolar endomitosis, nuclear fusion, and karyokinesis without cytokinesis. PGCCs underwent restitution multipolar endomitosis, nuclear fragmentation, and micronuclei formation to increase nuclear contents and heterogeneity. At the cellular level, the development of PGCCs was associated with forming transient intracellular cells, termed fecundity cells. The fecundity cells can be decellularized to facilitate nuclear fusion and synchronized with other nuclei for subsequent nuclear replication. PGCCs can undergo several rounds of entosis to form complex tissue structures, termed fecundity structures. The formation of PGCCs via multiple modes of nuclear replication in the absence of cytokinesis leads to an increase in the nuclear-to-cytoplasmic (N/C) ratio and intracellular cell reproduction, which is remarkably similar to the mode of nuclear division during pre-embryogenesis. Our data support that PGCCs may represent a central regulator in malignant histogenesis, intratumoral heterogeneity, immune escape, and macroevolution via the de-repression of suppressed pre-embryogenic program in somatic cells.

Malaria parasites harness Rho GTPase signaling and host cell membrane ruffling for productive invasion of hepatocytes

Plasmodium sporozoites are the motile forms of the malaria parasites that infect hepatocytes. The initial invasion of hepatocytes is thought to be actively driven by sporozoites, but host cell processes might also play a role. Sporozoite invasion triggers a host plasma membrane invagination that forms a vacuole around the intracellular parasite, which is critical for subsequent intracellular parasite replication. Using fast live confocal microscopy, we observed that the initial interactions between sporozoites and hepatocytes induce plasma membrane ruffles and filopodia extensions. Importantly, we find that these host cell processes facilitate invasion and that Rho GTPase signaling, which regulates membrane ruffling and filopodia extension, is critical for productive infection. Interestingly, sporozoite cell traversal stimulates these processes, suggesting that it increases hepatocyte susceptibility to productive infection. Our study identifies host cell signaling events involved in plasma membrane dynamics as a critical host component of successful malaria parasite infection of hepatocytes.

Umifenovir Epigenetically Targets the IL-10 Pathway in Therapy against Coxsackievirus B4 Infection.

Umifenovir, a broad-spectrum nonnucleoside antiviral drug, has a promising efficacy against coxsackievirus B4 (CVB4) infection, but its mechanism remains unclear. CVB4 is a common human single-stranded RNA virus that belongs to the Picornaviridae family and the Enterovirus genus. Enterovirus can cause severe diseases, such as meningitis, myocarditis, pancreatitis, insulin-dependent diabetes, and several other diseases, in both adults and children. We have previously demonstrated the critical role of interleukin 10 (IL-10) in promoting CVB4 infection and the downregulation of IL-10 by umifenovir. To further explore the underlying mechanisms of umifenovir, we characterized the epigenetic regulation of IL-10 in IL-10 knockout RAW264.7 cells and a BALB/c mouse splenocyte model. Mechanistically, we found that umifenovir inhibited CVB4-activated IL-10 by enhancing the methylation level of the repressive histones H3K9me3 and H3K27me3 while reducing the acetylation level of the activating histone H3K9ac in the promoter region of the IL-10 gene. Furthermore, using a chromosome conformation capture approach, we discovered that CVB4 infection activated the IL-10 gene by forming an intrachromosomal interaction between the IL-10 gene promoter and an intronic enhancer of the downstream MK2 (mitogen-activated protein kinase [MAPK]-activated protein kinase 2 [MAPKAPK2]) gene, a critical component of the p38-MAPK signaling pathway, which is required for IL-10 gene expression. However, umifenovir treatment abolished this spatial conformation and chromatin interaction, thus reducing the continuous expression of IL-10 and subsequent CVB4 replication. In conclusion, this study reveals a novel epigenetic mechanism by which umifenovir controls CVB4 infections, thus laying a theoretical foundation for therapeutic use of umifenovir. IMPORTANCE Viral infections are major threats to human health because of their strong association with a variety of inflammation-related diseases, especially cancer. Many antiviral drugs are performing poorly in treating viral infections. This is probably due to the immunosuppressive effect of highly expressed IL-10, which is caused by viral infection. Umifenovir is a broad-spectrum antiviral drug. Our recent studies showed that umifenovir has a significant inhibitory effect on CVB4 infection and can reduce IL-10 expression caused by CVB4. However, another antiviral drug, rupintrivir, showed good antiviral activity but had no effect on the expression of IL-10. This suggests that the regulation of IL-10 expression is a key part of the antiviral mechanism of umifenovir. Therefore, due to the dual function of the inhibition of CVB4 replication and the regulation of immune antiviral pathway, the mechanism of umifenovir is of great value to study.

Nonmuscle myosin IIA promotes the internalization of influenza A virus and regulates viral polymerase activity through interacting with nucleoprotein in human pulmonary cells

Influenza A virus (IAV), responsible for seasonal epidemics and recurring pandemics, represents a global threat to public health. Given the risk of a potential IAV pandemic, it is increasingly important to better understand virus-host interactions and develop new anti-viral strategies. Here, we reported nonmuscle myosin IIA (MYH9)-mediated regulation of IAV infection. MYH9 depletion caused a profound inhibition of IAV infection by reducing viral attachment and internalization in human lung epithelial cells. Surprisingly, overexpression of MYH9 also led to a significant reduction in viral productive infection. Interestingly, overexpression of MYH9 retained viral attachment, internalization, or uncoating, but suppressed the viral ribonucleoprotein (vRNP) activity in a minigenome system. Further analyses found that excess MYH9 might interrupt the formation of vRNP by interacting with the viral nucleoprotein (NP) and result in the reduction of the completed vRNP in the nucleus, thereby inhibiting subsequent viral RNA transcription and replication. Together, we discovered that MYH9 can interact with IAV NP protein and engage in the regulation of vRNP complexes, thereby involving viral replication. These findings enlighten new mechanistic insights into the complicated interface of host-IAV interactions, ultimately making it an attractive target for the generation of antiviral drugs.

A systematic review of the theory of mind studies and the potential for replications in linguistics

The theory of mind is a psychological concept that refers to the awareness of one’s mental states and others’. In the seminal study conducted by Kidd and Castano (2013), the researchers found that reading literary fiction led to better performance in the theory of mind test relative to reading popular fiction, nonfiction, or reading nothing at all. This paper collates seven subsequent replications of the original study: five direct replications and two indirect ones using other media, namely the visual narrative of television and movies. The findings of these replications are then compared to the original findings in terms of whether they achieved successful replication of the results as well. After that, key differences between the genres of the media used in the original study and its replications are elaborated, namely the writerly and readerly texts. For discussion, this paper further argues the case for replicating the original study in linguistics. In particular, this paper explores two main points pertaining to second language acquisition and emotions. The first one is that the original study and its replications were carried out exclusively with native speakers of English. Accordingly, this necessitates replicating the original study with participants other than first speakers. Meanwhile, considering the call for more replications, this paper makes the argument that since songs are short and also elicit emotions, they present another viable medium for indirect replications.

In Vitro Bactericidal Activity of An Environmentally Isolated Phage Cocktail Against a Characterized Recombinant Endolysin Lysecd7 on Clinical Bacterial Isolates

Background: Antimicrobial resistance (AMR) is rapidly spreading, which is pushing humanity into a “post-antibiotic era” where most antibiotics are no longer effective. Scientists are researching bacteriophages as a potential therapy against ESKAPE pathogens, which are a problem in hospital settings, as a result of the development of bacterial resistance to antibiotics. This study assessed the antibacterial efficacy of recombinant LysECD7 and environmental bacteriophage cocktail on bacterial isolates. Materials and Methods: Environmental water and soil samples were examined for the presence of bacteriophages that were capable of killing the bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterobacter cloacae. Utilizing environmental isolates, a bacteriophage cocktail was created and its ability to effectively lyse bacterial growth cultures in vitro was examined. A synthetically generated Myoviridae lytic enzyme named LysECD7 was cloned, produced, and demonstrated wide bactericidal efficacy against gram-negative bacterial pathogens. The bacteriophage cocktail and endolysin’s (0.203mg/ml) antibacterial activity were assessed using a spot assay and an antibacterial activity assay in broth culture. Phage and endolysin bacteria samples were taken and their OD600 levels were checked every 30 minutes for phages and every 30 seconds for endolysin. Results: The average antibacterial activity of isolated bacteriophages was 43.58%, as opposed to the recombinant endolysin’s 31.48%. Bacterial absorbance declined steadily with each subsequent phage replication cycle, however, after six hours, there was a minor improvement in bacterial growth. The Gram-negative E. coli ATCC 25922 and E. cloacae NCTC 134565 exhibited the highest activity for both treatments, while the S. aureus and P. aeruginosa had the lowest activity. Conclusion: A single dose of low endolysin concentration had a very significant antibacterial activity, bacteriophage treatment of bacterial isolates varied greatly per each species under the same conditions. Under the same circumstances, bacteriophage and endolysin activity change within bacterial isolates, demonstrating that activity is independent of concentration. Lysin activity was highest in E. coli and lowest in the other bacterial species, but it should be emphasized that lysin and bacteriophage activity and efficacy differ with each treatment.f

Unscheduled DNA replication in G1 causes genome instability and damage signatures indicative of replication collisions

DNA replicates once per cell cycle. Interfering with the regulation of DNA replication initiation generates genome instability through over-replication and has been linked to early stages of cancer development. Here, we engineer genetic systems in budding yeast to induce unscheduled replication in a G1-like cell cycle state. Unscheduled G1 replication initiates at canonical S-phase origins. We quantifiy the composition of replisomes in G1- and S-phase and identified firing factors, polymerase α, and histone supply as factors that limit replication outside S-phase. G1 replication per se does not trigger cellular checkpoints. Subsequent replication during S-phase, however, results in over-replication and leads to chromosome breaks and chromosome-wide, strand-biased occurrence of RPA-bound single-stranded DNA, indicating head-to-tail replication collisions as a key mechanism generating genome instability upon G1 replication. Low-level, sporadic induction of G1 replication induces an identical response, indicating findings from synthetic systems are applicable to naturally occurring scenarios of unscheduled replication initiation.

Legionella pneumophila: The Journey from the Environment to the Blood.

An outbreak of a potentially fatal form of pneumonia in 1976 and in the annual convention of the American Legion was the first time that Legionella spp. was identified. Thereafter, the term Legionnaires’ disease (LD) was established. The infection in humans is transmitted by the inhalation of aerosols that contain the microorganisms that belong to the Legionellaceae family and the genus Legionella. The genus Legionella contains genetically heterogeneous species and serogroups. The Legionella pneumophila serogroup 1 (Lp1) is the most often detected strain in outbreaks of LD. The pathogenesis of LD infection initiates with the attachment of the bacterial cells to the host cells, and subsequent intracellular replication. Following invasion, Legionella spp. activates its virulence mechanisms: generation of specific compartments of Legionella-containing vacuole (LCV), and expression of genes that encode a type IV secretion system (T4SS) for the translocation of proteins. The ability of L. pneumophila to transmigrate across the lung’s epithelium barrier leads to bacteremia, spread, and invasion of many organs with subsequent manifestations, complications, and septic shock. The clinical manifestations of LD depend on the bacterial load in the aerosol, the virulence factors, and the immune status of the patient. The infection has two distinct forms: the non- pneumatic form or Pontiac fever, which is a milder febrile flu-like illness, and LD, a more severe form, which includes pneumonia. In addition, the extrapulmonary involvement of LD can include heart, brain, abdomen, and joints.

Demystifying the components of public service motivation among young public servants in China: a qualitative inquiry

ABSTRACT While public service motivation (PSM) is universally acknowledged as a value-laden and context-contingent construct, few studies have relied on qualitative methodologies to investigate unique components and dimensions of PSM in a Chinese context. Thus, our knowledge of on-the-job motivation (held by Chinese public employees) and how the cultural and political systems shape these motivations remains limited. This study fills this gap through in-depth, semi-structured interviews with 22 junior public servants working at various organisational levels, geographic locations, and functional departments. The findings show that an attachment to the governance regime and predilection towards becoming a politician are two key dimensions that capture the rational motives among Chinese civil servants. In terms of norm-based motives, Confucianism-oriented authoritative values (including government-centric conception, collectivism, obedience, and moral obligations to serve the public) combine to affect the ways in which Chinese nascent public workers interpret the notion of public interests. In addition, love for both the family and nation serves as a core affective factor that contributes towards one’s willingness to seek membership in the public sector. This study ultimately helps construct an all-encompassing yet localised PSM concept, laying the foundation for subsequent quantitative examinations, validations, and replications of PSM-related theories in China.

Long-Term Effects of Kindergarten Vocabulary Instruction and Intervention on Target Vocabulary Knowledge Through Second Grade

We evaluated the long-term effects of a supplemental, small-group kindergarten vocabulary intervention in fall and spring of first grade and winter of second grade. Participants included students from two studies, an initial efficacy study and a subsequent replication study, identified as at risk for language and learning difficulties, who were randomly assigned in clusters to either a control group that received only classroom vocabulary instruction or a treatment group that received the classroom instruction plus small-group supplemental intervention. We also identified a group of not-at-risk students who received classroom vocabulary instruction as a reference group. Analyses using multilevel modeling indicated that students in the treatment group continued to outperform students in the control group on measures of expressive and receptive knowledge of words taught during the intervention through the winter of second grade, almost 2 years after the end of the intervention. Although long-term effects of the intervention on target word learning in second grade were substantial and meaningful, there was some deterioration of intervention effects across time. There were no effects of the kindergarten intervention on general vocabulary knowledge at posttest or any follow-up points. Findings suggest that the effects of direct and extended vocabulary instruction and intervention are generally durable and sustain over time for taught words.

Enhancement of oil repellency on hyperbolic microarrays by compressive bending of elastomeric films

Applying elastic deformation to modulate the microscale surface shapes of films is an efficient way to prepare samples with various geometric designs. Tensile stress can be applied by stretching films, and it can increase the pitches of line arrays or the diameters of pillar arrays on a film surface. However, it is difficult to apply lateral compressive stress to films because films buckle during compression. In this study, we exploit bending to apply lateral compressive stress to the surface of a film. To accomplish this, we use arrays of microhyperbolic pillars or lines that have inversely tapered shapes in the upper region and widened bottoms. By using deformation of elastomeric films and a subsequent replication process, we fabricate microhyperbolic structures with decreased pitches and increased pattern heights. In addition, the deformation of microarrays by compressive stress can enhance oil repellency by increasing the sharpness of the inversely tapered shapes. Using this concept, we demonstrate the enhancement of oil repellency on patterned surfaces prepared by compressive bending of microarrays. Furthermore, we provide selective oil repellency by modulating the compressive stress on a single line array. The concept of compressive bending can be applied widely to decrease the dimensions of arrays and modulate surface shapes for greater oil repellency.

A DNA replication fork-centric view of the budding yeast DNA damage response

The DNA damage response (DDR) checkpoint is activated when DNA is damaged or when DNA replication forks stall. The DDR checkpoint plays a critical role in preserving the integrity of stalled replication forks; this is essential for subsequent fork resumption, faithful and complete genome replication, and cell survival. The mechanisms by which the DDR checkpoint preserves stalled replication forks are still incompletely understood. Many substrates of the DDR checkpoint kinases have been identified over the years, but in many cases the functional consequences of phosphorylation are still unclear. Emerging as a complementary approach, recent advances in biochemical reconstitution of DNA replication have made it possible to characterise specific mechanisms of DNA replication regulation by the DDR checkpoint. In this review, we discuss the role of DNA replication in the activation of the DDR checkpoint and how this checkpoint regulates different aspects of DNA replication. We then distinguish between checkpoint action locally at the site of replication stalling and more globally, and we discuss how these functions contribute to coordinating complete replication of the genome in the face of replication stress.

Labor epidural analgesia and subsequent risk of offspring autism spectrum disorder and attention-deficit/hyperactivity disorder: a cross-national cohort study of 4.5 million individuals and their siblings

A recent study has suggested that labor epidural analgesia may be associated with increased rates of offspring autism spectrum disorder. Subsequent replication attempts have lacked sufficient power to confidently exclude the possibility of a small effect, and the causal nature of this association remains unknown. This study aimed to investigate the extent to which exposure to labor epidural analgesia is associated with offspring autism spectrum disorder and attention-deficit/hyperactivity disorder following adjustments for unmeasured familial confounding. We identified 4,498,462 singletons and their parents using the Medical Birth Registers in Finland (cohorts born from 1987-2005), Norway (1999-2015), and Sweden (1987-2011) linked with population and patient registries. These cohorts were followed from birth until they either had the outcomes of interest, emigrated, died, or reached the end of the follow-up (at mean ages 13.6-16.8 years), whichever occurred first. Cox regression models were used to estimate country-specific associations between labor epidural analgesia recorded at birth and outcomes (eg, at least 1 secondary care diagnosis of autism spectrum disorder and attention-deficit/hyperactivity disorder or at least 1 dispensed prescription of medication used for the treatment of attention-deficit/hyperactivity disorder). The models were adjusted for sex, birth year, birth order, and unmeasured familial confounders via sibling comparisons. Pooled estimates across all the 3 countries were estimated using inverse variance weighted fixed-effects meta-analysis models. A total of 4,498,462 individuals (48.7% female) were included, 1,091,846 (24.3%) of which were exposed to labor epidural analgesia. Of these, 1.2% were diagnosed with autism spectrum disorder and 4.0% with attention-deficit/hyperactivity disorder. On the population level, pooled estimates showed that labor epidural analgesia was associated with increased risk of offspring autism spectrum disorder (adjusted hazard ratio, 1.12; 95% confidence interval, 1.10-1.14, absolute risks, 1.20% vs 1.07%) and attention-deficit/hyperactivity disorder (adjusted hazard ratio, 1.20; 95% confidence interval, 1.19-1.21; absolute risks, 3.95% vs 3.32%). However, when comparing full siblings who were differentially exposed to labor epidural analgesia, the associations were fully attenuated for both conditions with narrow confidence intervals (adjusted hazard ratio [autism spectrum disorder], 0.98; 95% confidence interval, 0.93-1.03; adjusted hazard ratio attention-deficit/hyperactivity disorder, 0.99; 95% confidence interval, 0.96-1.02). In this large cross-national study, we found no support for the hypothesis that exposure to labor epidural analgesia causes either offspring autism spectrum disorder or attention-deficit/hyperactivity disorder.

MORC3 restricts human cytomegalovirus infection by suppressing the major immediate-early promoter activity.

During the long coevolution of human cytomegalovirus (HCMV) and humans, the host has formed a defense system of multiple layers to eradicate the invader, and the virus has developed various strategies to evade host surveillance programs. The intrinsic immunity primarily orchestrated by promyelocytic leukemia (PML) nuclear bodies (PML-NBs) represents the first line of defense against HCMV infection. Here, we demonstrate that microrchidia family CW-type zinc finger 3 (MORC3), a PML-NBs component, is a restriction factor targeting HCMV infection. We show that depletion of MORC3 through knockdown by RNA interference or knockout by CRISPR-Cas9 augmented immediate-early protein 1(IE1) gene expression and subsequent viral replication, and overexpressing MORC3 inhibited HCMV replication by suppressing IE1 gene expression. To relief the restriction, HCMV induces transient reduction of MORC3 protein level via the ubiquitin-proteasome pathway during the immediate-early to early stage. However, MORC3 transcription is upregulated, and the protein level recovers in the late stages. Further analyses with temporal-controlled MORC3 expression and the major immediate-early promoter (MIEP)-based reporters show that MORC3 suppresses MIEP activity and consequent IE1 expression with the assistance of PML. Taken together, our data reveal that HCMV enforces temporary loss of MORC3 to evade its repression against the initiation of immediate-early gene expression.