Genome-wide loss-of-function genetic screen identifies INSIG2 as the vulnerability of hepatitis B virus-integrated hepatoma cells.

There are approximately 250 million people chronically infected with hepatitis B virus (HBV) worldwide. Although HBV is often integrated into the host genome and promotes hepatocarcinogenesis, vulnerability of HBV integration in liver cancer cells has not been clarified. The aim of our study is to identify vulnerability factors for HBV-associated hepatocarcinoma. Loss-of-function screening was undertaken in HepG2 and HBV-integrated HepG2.2.15 cells expressing SpCas9 using a pooled genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) library. Genes whose guide RNA (gRNA) abundance significantly decreased in HepG2.2.15 cells but not in HepG2 cells were extracted using the MAGeCK algorithm. We identified four genes (BCL2L1, VPS37A, INSIG2, and CFLAR) that showed significant reductions of gRNA abundance and thus potentially involved in the vulnerability of HBV-integrated cancer cells. Among them, siRNA-mediated mRNA inhibition or CRISPR-mediated genetic deletion of INSIG2 significantly impaired cell proliferation in HepG2.2.15 cells but not in HepG2 cells. Its inhibitory effect was alleviated by cotransfection of siRNAs targeting HBV. INSIG2 inhibition suppressed the pathways related to cell cycle and DNA replication, downregulated cyclin-dependent kinase 2 (CDK2) levels, and delayed the G1 -to-S transition in HepG2.2.15 cells. CDK2 inhibitor suppressed cell cycle progression in HepG2.2.15 cells and INSIG2 inhibition did not suppress cell proliferation in the presence of CDK2 inhibitor. In conclusion, INSIG2 inhibition induced cell cycle arrest in HBV-integrated hepatoma cells in a CDK2-dependent manner, and thus INSIG2 might be a vulnerability factor for HBV-associated liver cancer.

Collaborative Mother-Toddler Communication and Theory of Mind Development at Age 4.

Focusing on social pragmatics, this longitudinal study investigated the contribution of mother-toddler collaborative communication to theory of mind (ToM) development at age 4. At age 2½, 78 toddlers' (42 boys) and their mothers were observed during pretend play. At age 4, children were tested using 4 false belief understanding tasks. Both mothers and toddlers engaged in more collaborative (inform, guide/request, and support/confirm) than non-collaborative communication acts. Other-focused collaborative acts of support/confirm by mothers and toddlers predicted children's false belief understanding, even after controlling for 5 covariates. In addition, as active agents in their own ToM development, the contribution of toddlers' collaborative acts to false belief understanding was independent of their mothers. Finally, the way toddlers and their mothers co-constructing their communication mattered. Only when toddlers engaged in high levels of collaborative acts, the mothers' high levels of collaborative acts demonstrated a positive effect on children's ToM development. The applied implications of these findings were discussed.

The serum tenascin C level is a marker of metabolic disorder-related inflammation affecting pancreatic cancer prognosis.

Obesity is a risk factor for pancreatic cancer development, partly due to the tissue environment of metabolic disorder-related inflammation. We aimed to detect a tissue environment marker triggered by obesity-related metabolic disorders related to pancreatic cancer progression. In murine experiments, Bl6/j mice fed a normal diet (ND) or a high-fat diet (HFD) were orthotopically injected with mPKC1, a murine-derived pancreatic cancer cell line. We used stocked sera from 140 pancreatic cancer patients for analysis and 14 colon polyp patients as a disease control. Compared with ND-fed mice, HFD-fed mice exhibited obesity, larger tumors, and worse prognoses. RNA sequencing of tumors identified tenascin C (TNC) as a candidate obesity-related serum tissue environment marker with elevated expression in tumors of HFD-fed mice. Serum TNC levels were greater in HFD-fed mice than in ND-fed mice. In pancreatic cancer patients, serum TNC levels were greater than those in controls. The TNC-high group had more metabolic disorders and greater CA19-9 levels than did the TNC-low group. There was no relationship between serum TNC levels and disease stage. Among 77 metastatic patients treated with chemotherapy, a high serum TNC concentration was an independent poor prognostic factor. Pancreatic cancer patients with high serum TNC levels experienced progression more rapidly.

Microstructural Control Strategy Based on Optimizing Laser Powder Bed Fusion for Different Hastelloy X Powder Size.

In additive manufacturing (AM), the powder properties and laser powder bed fusion (LPBF) process parameters influence the quality of materials and building parts. However, the relationship between the size of the powder, LPBF process parameters, and mechanical properties is not well-established. In addition, Hastelloy X (HX) is attracting attention for its excellent high-temperature properties, but it is difficult to process, such as by cutting and milling, because of its high hardness and high ductility. This can be overcome by applying the AM process. We compared the LPBF window process maps for two HX powders of different sizes. Despite their small difference of 19.7% in particle size, it was confirmed that the difference in laser power was more than 40 W, the difference in scan speed was more than 100 mm/s, and the difference in energy density was more than 20% under the optimal process conditions. The as-built specimen had a larger molten-pool size as the energy density was higher, which resulted in the differences in mechanical properties at room temperature and high temperature (816 °C). We considered the control of the size of the powder to obtain the properties required for each temperature condition. The microstructures and mechanical properties of the as-built LPBF specimens were also investigated and compared with those of cast HX. Because of the rapid melting and solidification processes in LPBF, the as-built HX exhibited nano-sized dendrite structures and large internal strain energy. This resulted in the as-built LPBF exhibiting a higher room-temperature tensile strength than the cast material. Under high-temperature conditions, the grain boundary of the as-built LPBF acts as a sliding path, and the as-built LPBF HX showed significantly better high-temperature tensile strength characteristics than the cast HX.

Exploring the use and challenges of implementing virtual visits during COVID-19 in primary care and lessons for sustained use.

BACKGROUND: The COVID-19 pandemic has rapidly transformed how healthcare is delivered to limit the transmission of the virus. This descriptive cross-sectional study explored the current use of virtual visits in providing care among primary care providers in southwestern Ontario during the first wave of the COVID-19 pandemic and the anticipated level of utilization post-pandemic. It also explored clinicians' perceptions of the available support tools and resources and challenges to incorporating virtual visits within primary care practices. METHODS: Primary care physicians and nurse practitioners currently practicing in the southwestern part of Ontario were invited to participate in an online survey. The survey invite was distributed via email, different social media platforms, and newsletters. The survey questions gathered clinicians' demographic information and assessed their experience with virtual visits, including the proportion of visits conducted virtually (before, during the pandemic, and expected volume post-pandemic), overall satisfaction and comfort level with offering virtual visits using modalities, challenges experienced, as well as useful resources and tools to support them in using virtual visits in their practice. RESULTS: We received 207 responses, with 96.6% of respondents offering virtual visits in their practice. Participants used different modalities to conduct virtual visits, with the vast majority offering visits via phone calls (99.5%). Since the COVID-19 pandemic, clinicians who offered virtual visits have conducted an average of 66.4% of their visits virtually, compared to an average of 6.5% pre-pandemic. Participants anticipated continuing use of virtual visits with an average of 43.9% post-pandemic. Overall, 74.5% of participants were satisfied with their experience using virtual visits, and 88% believed they could incorporate virtual visits well within the usual workflow. Participants highlighted some challenges in offering virtual care. For example, 58% were concerned about patients' limited access to technology, 55% about patients' knowledge of technology, and 41% about the lack of integration with their current EMR, the increase in demand over time, and the connectivity issues such as inconsistent Wi-Fi/Internet connection. There were significant differences in perception of some challenges between clinicians in urban vs, rural areas. Clinicians in rural areas were more likely to consider the inconsistent Wi-Fi and limited connectivity as barriers to incorporating virtual visits within the practice setting (58.8% vs. 40.2%, P = 0.030). In comparison, clinicians in urban areas were significantly more concerned about patients overusing virtual care services (39.4% vs. 21.6%, P = 0.024). As for support tools, 47% of clinicians advocated for virtual care standards outlined by their profession's college. About 32% identified change management support and technical training as supportive tools. Moreover, 39% and 28% thought local colleagues and in-house organizational support are helpful resources, respectively. CONCLUSION: Our study shows that the adoption of virtual visits has exponentially increased during the pandemic, with a significant interest in continuing to use virtual care options in the delivery of primary care post-pandemic. The study sheds light on tools and resources that could enhance operational efficiencies in adopting virtual visits in primary care settings and highlights challenges that, when addressed, can expand the health system capacity and sustained use of virtual care.

Water-slider method reduced colonic polyp fragmentation compared with conventional suction retrieval: A prospective randomized controlled study.

OBJECTIVES: After resecting colonic polyps, retrieval through the scope channel may lead to polyp fragmentation, making pathologic evaluation challenging. An easy and reliable method for complete polyp retrieval is needed. METHODS: We developed the water-slider method (WSM), in which the resected polyp is surrounded by water from an auxiliary water channel during suctioning. We prospectively randomized patients who underwent endoscopic resection for colonic polyps in our institute into WSM and non-WSM groups, and evaluated the polyp fragmentation rate. RESULTS: Analysis of the data regarding small polyps (≤10 mm in size) revealed that the WSM group had a significantly lower polyp fragmentation rate (8.2%) than the non-WSM group (23.8%, p < 0.001). Polyp retrieval time did not differ significantly between groups. The rate of a clear-cut end on neoplastic polyps was significantly higher in the WSM group (63.8%) than in the non-WSM group (50.0%; p = 0.029). CONCLUSIONS: The WSM achieved a significantly lower polyp fragmentation rate, allowing for more accurate histologic evaluation than the conventional method. The WSM did not influence the polyp retrieval time.

A Study on the Mechanical Properties of an Automobile Part Additively Printed through Periodic Layer Rotation Strategies.

In metal product manufacturing, additive manufacturing (AM) is a method that has the advantage of fabricating complex shapes and customized production, unlike existing machining methods. However, owing to the characteristics of the AM process, anisotropy of macrostructure occurs because of various causes such as the scan direction, melting, fusion, and cooling of the powdered material. The macrostructure anisotropy is realized from the scan direction, and when a single layer is stacked in one direction, it is expressed as orthogonal anisotropy. Here, the classical lamination theory is applied to simply calculate the individual orthotropic layers by superimposing them. Through this, the authors analyzed whether the mechanical properties of the product are isotropically expressed with a periodic layer rotation strategy. To determine if the mechanical properties can be reasonably considered to be isotropic, a shock absorber mount for a vehicle was manufactured by AM. The tensile and vibration test performed on the product was compared with the finite element analysis and experimental results. As a result of the comparison, it was confirmed that the macroscopically of the product was considered isotropic as the load-displacement diagram and the fracture location coincided, as well as the natural frequency and mode shape.

Properties of Tool Steels Printed by Directed Energy Deposition Process on S45C Base Metal.

We present a fundamental study on the development of trimming dies at room temperature for the hot-stamping process using directed energy deposition. Specimens of G and F materials were fabricated by machining 3D-printed blocks. The hardness of G-layered specimens was slightly higher than that of F-layered specimens, reaching approximately 700 HV at the surface. The G-layered specimens consisted of columnar and equiaxed dendrites, whereas the F-layered specimens mainly consisted of equiaxed dendrites. Spherical pores were observed inside the layered cross section, whereas relatively large irregular-shaped cavities were observed in layered boundaries. The tensile strengths of the G-layered and F-layered specimens were approximately 1800 and 1650 MPa, respectively. During bonding strength tests on an area bonded with S45C base metal, a fracture occurred in one case because of the lack of fusion at the boundary, and the F-layered specimens showed a lower strength than the G-layered ones. During wear tests on a quenched 1.5 GPa-grade aluminized steel plate, the F-layered specimens showed lower wear loss. However, the G-layered specimens showed better wear resistance during wear tests on a 1.5 GPa-grade electrogalvanized steel plate. These findings serve as fundamental data for additive manufacturing processes using tool steels of high-strength materials with high melting points.

Multimodal sex-related differences in infant and in infant-directed maternal behaviors during months three through twelve of development.

Using the concepts of sensory and affective experience, this work relates the concepts of socialization and cognitive development to the embodiment of gender in the human infant. Evidence obtained from biweekly observations from 30 children and their mothers observed from age 3 months to age 12 months revealed measurable sex-related differences in how mothers handle and touch their infants. This work offers novel approaches to visualizing combinations of behaviors with the aim of encouraging researchers to think in terms of suites of action rather than singular sensory or motor systems. New avenues of research into the mechanisms which produce sex-related differences in behavior are suggested.

Improved sero-monitoring assay for classical swine fever (CSF) using the recombinant E2 protein of a recent Korean isolate.

Classical swine fever (CSF) is a highly contagious disease of pigs that causes fever, diarrhea and paralysis, often resulting in death. E2 is the major structural protein of the CSF virus (CSFV) and mediates the entrance of the virus, subsequently inducing a neutralizing immune response. In this study, the E2 gene of a recent Korean isolate of CSF, SW03, was cloned and the DNA sequence was compared to other strains via phylogenetic analysis. With the purified E2 protein, an enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of CSFV infection. The sensitivity and specificity of the E2-ELISA were 96.1% and 94.8%, respectively. A total of 17 out of 485 field-collected pig sera tested demonstrated conflicting results between two ELISA methods, a commercial kit and the E2-ELISA. Of these sera, 60% were determined to be CSFV positive by a virus neutralization test (VNT), suggesting involvement of different immune responses in the cases of CSFV infection. As the E2-ELISA was developed using a recent Korean isolate, SW03, this assay is capable of rapidly identifying newly emerging CSFV strains.

First-line gefitinib treatment for patients with advanced non-small cell lung cancer with poor performance status.

BACKGROUND: Best supportive care only is recommended for patients with advanced non-small cell lung cancer (NSCLC) with poor performance status (PS) of Eastern Cooperative Oncology Group 3 or 4. Recently, the possibility of using epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor therapy has been reported for poor PS patients harboring EGFR mutations. METHODS: We retrospectively analyzed 74 patients with advanced NSCLC who were treated with first-line gefitinib during hospitalization for Eastern Cooperative Oncology Group PS 3 or 4. All patients were classified according to three clinical parameters: smoking history, gender, and histology type. RESULTS: The median age was 64 years (range, 35-86 years). The proportions of females, never smokers, and adenocarcinoma were 51.4%, 54.1%, and 78.4%, respectively. An overall response rate, median progression-free survival (PFS), and median overall survival (OS) was 27.0%, 32 days (95% confidence interval [CI], 22-48 days), and 61 days (95% CI, 7-115 days), respectively. Female gender, never smoking, and adenocarcinoma histology were strong predictors of tumor response. Never smoking and adenocarcinoma were independent predictors of better PFS but not of OS. Seven patients experienced treatment-related adverse effects of grade 3 to 4, which included anorexia (n = 2), pneumonitis (n = 4), and elevated liver enzymes (n = 1). Never-smoker females with adenocarcinoma exhibited a response rate of 50.0%, median PFS of 130 days (95% CI, 51-209 days), and median OS of 236 days (95% CI, 150-322 days). CONCLUSIONS: Gefitinib may provide clinical benefits for patients with NSCLC with poor PS who were selected according to clinically favorable parameters.

Separation anxiety in first-time mothers: infant behavioral reactivity and maternal parenting self-efficacy as contributors.

This short-term longitudinal study examined the contribution of infant behavioral reactivity and maternal parenting self-efficacy to first-time mothers' (total n=32) separation anxiety. Infants' behavioral reactivity indexed by gaze and facial affect were observed in the conditions of routine play and maternal still face at 3 months. Mothers reported their self-perceived parenting efficacy at 3 months and separation anxiety at 6 months. Results revealed that infants' frequent gazing at their mother, greater negative affect, and less positive affect observed during the still face, but not during the routine play, contributed to heightened maternal separation anxiety. In addition to a direct link between low maternal parenting self-efficacy and high maternal separation anxiety, maternal parenting self-efficacy buffered against the impact of infant negative reactivity on maternal separation anxiety. The role of infant reactivity and maternal self-efficacy in parenting was discussed.