A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in VeroE6 cells

We developed a multi-scale microscopy imaging toolbox to address some major issues related to SARS-CoV-2 interactions with host cells. Our approach harnesses both conventional and super-resolution fluorescence microscopy (Airyscan, STORM, and STED) and easily matches the spatial scale of single virus-cell checkpoints. We deployed this toolbox to characterize subtle issues related to the entry phase of SARS-CoV-2 variants in VeroE6 cells. Our results suggest that the variant of concern B.1.1.7, currently on the rise in several countries by a clear transmission advantage, in these cells outcompetes its ancestor B.1 in terms of a much faster kinetics of entry. Given the molecular scenario (entry by the late pathway and similar fraction of pre-cleaved S protein for B.1.1.7 and B.1), the faster entry of B.1.1.7 could be directly related to the N501Y mutation in the S protein, which is known to strengthen the binding of Spike RBD with ACE2. Remarkably, we also observed directly the significant role of clathrin as mediator of late entry endocytosis, which had been previously suggested in analogy with other CoVs and from experiments on pseudotyped virus models. On overall, we believe that our fluroescence microscopy-based approach is valuable for future studies addressing of how SARS-CoV-2 and its variants interact with cells.

Born-Again Globals

Based on the literature on the Uppsala model, born-again globals, non-linear internationalization model, and late market entry, this chapter aims to portray the history, changes, and adaptations of OMEGA's internationalization process. This transitioning firm manufactures furniture and wooden hockey sticks. This chapter identifies that OMEGA follows a non-linear internationalization process and late entry into international markets. The principal added value of the case study presented here is related to presenting OMEGA's non-linear internationalization process, which displays reactive internationalization behavior in response to a saturated domestic market, typical of the Uppsala model, and which subsequently ends in a rapid internationalization process, as a born-again global, as a result of a change in its top management. During its internationalization process, OMEGA changed its internationalization pace, modes of entry, and export actions by adapting to the external environment and then changing its strategic focus.

PERAN GURU BK DENGAN LAYANAN INFORMASI DALAM MENGATASI KENAKALAN REMAJA BAGI SISWA KELAS X DI SMK TUNAS BANGSA

Tujuan penelitian ini adalah untuk mengetahui layanan informasi yang berjalan saat ini dan untuk mengetahui peran layanan informasi dalam mengangani kenakalan remaja pada siswa kelas X SMK Tunas Bangsa. Desain penelitian yang digunakan yaitu metode kualitatif.. Adapun jumlah sampel yang dibutuhkan dalam penelitian ini dengan lima orang siswa yaitu dengan meminta bantuan guru BK memilih beberapa siswa untuk mewawancara dan memberikan layanan informasi kepada lima orang siswa. Teknik analisis data yang digunakan yaitu wawancara, dan observasi. Dari hasil penelitian dapat ditari kesimpulan layanan informasi yang deiberikan untuk memberikan pemahaman kepada siswa tentang kenakalan remaja terutama tentang informasi yang dibutuhkan dan bisa membantu siswa dalam mengambil keputusan yang tepat dan ditemukannya bentuk kenalakan remaja yang ada di SMK Tunas Bangsa yaitu terlambat masuk sekolah, membolos, menyontek dan melanggar peratusan sekolah. Sarannya untuk guru bimingan konseling diharapkan lebih memberikan informasi sebnyak-banyaknya kepada siswa terkait dengan kenakalan remaja. ___________________________________________________________________ The purpose of this study was to determine information services currently running and to determine the role of information services in dealing with juvenile delinquency in class X SMK Tunas Bangsa students. The research design used is a qualitative method. The number of samples needed in this study with five students, namely by asking the guidance counselor to select several students to interview and provide information services to five students. The data analysis techniques used were interviews and observation. From the research results can be drawn the conclusion of the information services provided to provide understanding to students about juvenile delinquency, especially about the information needed and can help students in making the right decisions and finding forms of adolescent acquaintance in SMK Tunas Bangsa, namely late entry to school, truancy, cheating and violating school requirements. The suggestion for counseling guidance teachers is that it is hoped that they will provide as much information as possible to students regarding juvenile delinquency.

Saving Some Inequality for Young Adulthood? Social Origin Differences in ‘Re’-enrollment in Postsecondary Education in the United States, Sweden, and Germany

In this article, three country contexts are compared with regard to inequality by social origin in late enrollment in post-secondary education. Countries differ in the frequency at which young adults leave the labor market and (re)-enroll. This article discusses how inequality in late entry into education by social origin can be linked to characteristics of the education system and the labor market. Two cases with education systems that welcome adult learners and offer many second chances – Sweden and the US – are contrasted with the German system before its recent reforms. Systems with a welcoming environment for adult learners in higher education are shown to have higher educational inequalities in later transitions, while inequality in earlier transitions is lower.

A Hyperstabilizing Mutation in the Base of the Ebola Virus Glycoprotein Acts at Multiple Steps To Abrogate Viral Entry

ABSTRACTEbola virus (EBOV) causes highly lethal disease outbreaks against which no FDA-approved countermeasures are available. Although many host factors exploited by EBOV for cell entry have been identified, including host cell surface phosphatidylserine receptors, endosomal cysteine proteases, and the lysosomal cholesterol trafficking protein NPC1, key questions remain. Specifically, late entry steps culminating in viral membrane fusion remain enigmatic. Here, we investigated a set of glycoprotein (GP) mutants previously hypothesized to be entry defective and identified one mutation, R64A, that abolished infection with no apparent impact on GP expression, folding, or viral incorporation. R64A profoundly thermostabilized EBOV GP and rendered it highly resistant to proteolysisin vitro. Forward-genetics and cell entry studies strongly suggested that R64A’s effects on GP thermostability and proteolysis arrest viral entry at least at two distinct steps: the first upstream of NPC1 binding and the second at a late entry step downstream of fusion activation. Concordantly, toremifene, a small-molecule entry inhibitor previously shown to bind and destabilize GP, may selectively enhance the infectivity of viral particles bearing GP(R64A) at subinhibitory concentrations. R64A provides a valuable tool to further define the interplay between GP stability, proteolysis, and viral membrane fusion; to explore the rational design of stability-modulating antivirals; and to spur the development of next-generation Ebola virus vaccines with improved stability.IMPORTANCEEbola virus is a medically relevant virus responsible for outbreaks of severe disease in western and central Africa, with mortality rates reaching as high as 90%. Despite considerable effort, there are currently no FDA-approved therapeutics or targeted interventions available, highlighting the need of development in this area. Host-cell invasion represents an attractive target for antivirals, and several drug candidates have been identified; however, our limited understanding of the complex viral entry process challenges the development of such entry-targeting drugs. Here, we report on a glycoprotein mutation that abrogates viral entry and provides insights into the final steps of this process. In addition, the hyperstabilized phenotype of this mutant makes it useful as a tool in the discovery and design of stability-modulating antivirals and next-generation vaccines against Ebola virus.

PERUBAHAN PERILAKU NEGATIF SISWA MELALUI PENERAPAN TEAM GAME TOURNAMENT (TGT) PADA MATERI AKHLAK TERPUJI DI KELAS VIII MTS AL-MUNAWWAROH - KAB. MALANG

The use of methods that are not in accordance with the purpose of teaching will be an obstacle in achieving the objectives that have been formulated. This study is to find out the changes in students' negative behavior through the application of cooperative methods team games tournament (TGT) models on the material of commendable morals to class VIII students at MTs Al-Munawwaroh - Kab. Poor. This research is a classroom action research that begins pre-cycle and is carried out with 2 (two) cycles. The research subjects were 26 students. The results showed that the TGT method in Aqidah Akhlak subjects was able to change students' negative behavior. This can be seen from the comparison starting from the pre-cycle to the second cycle, namely leaving the class without permission decreasing from 11.5% to 0%, often permission to go to the toilet when learning decreased from 31% to 8%, late entry decreased from 19% to 4%, annoying friends when learning decreases from 11.5% to 0%.

Counterpoint: Keeping the Demons at Bay When Handling Time-Varying Exposures—Beyond Avoiding Immortal Person-Time

The potential for immortal time bias is pervasive in epidemiologic studies with left truncation or time-varying exposures. Unlike other biases in epidemiologic research (e.g., measurement bias, confounding due to unmeasured factors, and selection based on unmeasured predictors of the outcome), immortal time bias can and should be avoided by the correct assignment of person-time during follow up. However, even when handing person-time correctly, allowing late entry into a study or into an exposure group can open the door to more insidious sources of bias, some of which we explore here. Clear articulation of the study question, including the treatment plans of interest, can provide navigation around these sources of bias and elucidate the assumptions needed for inference given the available data. Here, we use simulated data to illustrate the assumptions required under various approaches to estimate the effect of a time-varying treatment and describe how these assumptions relate to the assumptions necessary to estimate single sample rates and risks in settings with censoring and truncation.