In the midst of an unstable Middle East, occupied with a combination of authentic civil protests, and illegitimate ISIS attacks, Lebanon seems to be a promising spot to true reconciliation.
As global business continues to expand and bring everyone closer, the critical element of a successful business outcome may be the appreciation and respect for regional, country, and cultural differences - known as cultural diversity and requiring good intercultural communication.
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1. Beckon with index finger. This means Come here in the U.S. To motion with the index finger to call someone is insulting, or even obscene, in many cultures. Expect a reaction when you beckon to a student from the Middle or Far East; Portugal, Spain, Latin America, Japan, Indonesia and Hong Kong. It is more acceptable to beckon with the palm down, with fingers or whole hand waving.
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Absence of Trop2 impairsproliferation of compact-bone derived MSCs. (A) CCK-8 assay of MSCsafter different passages of the cultures. For both KO and WTderived MSCs at the designated passages, equal amounts of the cells(2000 cells/100 μl) were seeded in flat-bottomed 96-well plates ata final volume of 100 μl. The cell proliferation (total viable cellnumbers) in each well was estimated by optical density (450 nm)using a Multiscan microplate spectrophotometer (Thermo Scientific,Rockford, IL, USA) at 0, 24, 48, 72, and 96 h, after initialincubation for 3 h. Except for the passage 13 (the last passage),MSCs from Trop2 KO mice showed a significantly decreasedviable cell number than that of WT for the assays after 48 h ofculture (n=3, *P
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Absence of Trop2 impairsdifferentiation of compact-bone derived MSCs. (A) Adipogenesis andosteogenesis were impaired in Trop2 KO MSCs. Equal amountsof the cells from the KO and WT mice were cultured under theconditioned medium for 3–4 weeks, and the cell differentiation wasestimated by Red Oil O and Alizarin Red staining for adipogenesisand osteogenesis, respectively. The untreated MSCs from the KO andWT mice were found negative for both adipogenesis and osteogenesis(a and b). In contrast, a significantly higher percentage ofadipogenesis was observed in MSCs of WT (c) than that ofTrop2 KO (d). Similarly, osteogenesis differentiation inMSCs of WT (e) was significantly higher than that of KO (f). (B andC) mRNA expression levels measured by RT-qPCR on WT andTrop2 KO MSCs of the marker genes were specifically involvedin the process of adipogenesis (PPAR-γ2 and Adipson)and osteogenesis (Osteocalcin and Osteopontin) weresignificantly decreased in the MSCs of Trop2 KO mice (n=3;*P
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Trop2 deficiency impairs AKTactivation, Cyclin D1 expression, and cell cycle progression. (A)Cell cycle analysis of the MSCs cultures at different passages. WTand Trop2 KO mouse compact bone-derived MSCs after differentpassages of cultures were collected, fixed, stained with propidiumiodide and analyzed for cell cycle phase distribution. Comparedwith WT, the proportion of MSCs entering the S phase fromTrop2 KO mice was significantly reduced compared to that ofWT at the same passage (n=3, *P
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The result that the proportion of active AKT in MSCsfrom KO mice was significantly lower than that from WTmice suggested that deficiency impaired the activationof AKT. However, we noted that the ratio of p-AKT/AKT andexpression of cyclin D1 showed a tread of decline with theprogression of the MSC culture passage, and the same was true forTrop2 expression except in passage 1, which was affected byhematopoietic cell contamination. This may be attributed to thefact of no addition of cytokines for both promoting proliferationand maintaining of undifferentiated state of MSCs during oursubculture (,). Without addition of the cytokines,MSCs became senescent and lost the potential of self-renewal,accompanied by gradually reduced expression of Trop2 during thepassage. Thus, Trop2 may protect MSCs from aging during thepassage, while the absence of may promote MSCssenescence, which would subsequently impair the self-renewalpotential of MSCs. However, Trop2 loss may not completely block AKTactivation. On one hand, Trop2 deficiency cannot completely abolishAKT activation by some unknown cytokines in the MSC-qualified fetalbovine serum, or AKT may be activated in multiple pathways throughcomplex signal networks (). Onthe other hand, may have pleiotropic functions as wellas EpCAM, which regulates both adhesion (), and intracellular signaling viacleavage products (). Lateststudy revealed that Trop2 loss promoted the tumor aggressivenessand epithelial-mesenchymal trans-differentiation (); while previous studies emphasizedthat Trop2 expression was positively related to the tumoraggressiveness and metastasis. Hence, further studies are requiredto elucidate the relationships between Trop2 and related signalingpathways in MSCs or other types of stem cells.