The role of Importin-βs in the maintenance and lineage commitment of mouse embryonic stem cells

AbstractMembers of the Importin-β family recognize nuclear localization signals (NLS) and nuclear export signals (NES). These proteins play important roles in various nucleocytoplasmic transport processes in cells. Here, we examined the expression patterns of 21 identified Importin-β genes in mouse embryonic stem cells (mESCs), mouse embryonic fibroblast (MEF) and mESCs differentiated into neural ectoderm (NE) or mesoendoderm (ME). We observed striking differences in the Importin-β mRNA expression levels within these cell types. We also found that knockdown of selected Importin-β genes led to suppression of Nanog, and altered the balance of Oct4/Sox2 expression ratio, which is important for NE/ME lineage choice. Furthermore, we demonstrated that knockdown of XPO4, RanBP17, RanBP16, or IPO7 differentially affected the lineage selection of differentiating mESCs. More specifically, knockdown of XPO4 selectively stimulated the mESC differentiation towards definitive endoderm, while concomitantly inhibiting NE differentiation. RanBP17 knockdown also promoted endodermal differentiation with no effect on NE differentiation. RanBP16 knockdown caused differentiation into ME, while IPO7 knockdown inhibited NE differentiation, without obvious effects on the other lineages. Collectively, our results suggest that Importin-βs play important roles in cell fate determination processes of mESCs, such as in the maintenance of pluripotency or selection of lineage during differentiation

Contribution of acidic extracellular microenvironment of cancer-colonized bone to bone pain

Solid and hematologic cancer colonized bone produces a number of pathologies. One of the most common complications is bone pain. Cancer-associated bone pain (CABP) is a major cause of increased morbidity and diminishes the quality of life and affects survival. Current treatments do not satisfactorily control CABP and can elicit adverse effects. Thus, new therapeutic interventions are needed to manage CABP. However, the mechanisms responsible for CABP are poorly understood. The observation that specific osteoclast inhibitors can reduce CABP in patients indicates a critical role of osteoclasts in the pathophysiology of CABP. Osteoclasts create an acidic extracellular microenvironment by secretion of protons via vacuolar proton pumps during bone resorption. In addition, bone-colonized cancer cells also release protons and lactate via plasma membrane pH regulators to avoid intracellular acidification resulting from increased aerobic glycolysis known as the Warburg effect. Since acidosis is algogenic for sensory neurons and bone is densely innervated by sensory neurons that express acid-sensing nociceptors, the acidic bone microenvironments can evoke CABP. Understanding of the mechanism by which the acidic extracellular microenvironment is created in cancer-colonized bone and the expression and function of the acid-sensing nociceptors are regulated should facilitate the development of novel approaches for management of CABP. Here, the contribution of the acidic microenvironment created in cancer-colonized bone to elicitation of CABP and potential therapeutic implications of blocking the development and recognition of acidic microenvironment will be described. This article is part of a Special Issue entitled: Membrane channels and transporters in cancers

Two mechanisms of oral malodor inhibition by zinc ions

Objectives The aim of this study was to reveal the mechanisms by which zinc ions inhibit oral malodor. Material and Methods The direct binding of zinc ions to gaseous hydrogen sulfide (H2S) was assessed in comparison with other metal ions. Nine metal chlorides and six metal acetates were examined. To understand the strength of H2S volatilization inhibition, the minimum concentration needed to inhibit H2S volatilization was determined using serial dilution methods. Subsequently, the inhibitory activities of zinc ions on the growth of six oral bacterial strains related to volatile sulfur compound (VSC) production and three strains not related to VSC production were evaluated. Results Aqueous solutions of ZnCl2, CdCl2, CuCl2, (CH3COO)2Zn, (CH3COO)2Cd, (CH3COO)2Cu, and CH3COOAg inhibited H2S volatilization almost entirely. The strengths of H2S volatilization inhibition were in the order Ag+ >; Cd2+ >; Cu2+ >; Zn2+. The effect of zinc ions on the growth of oral bacteria was strain-dependent. Fusobacterium nucleatum ATCC 25586 was the most sensitive, as it was suppressed by medium containing 0.001% zinc ions. Conclusions Zinc ions have an inhibitory effect on oral malodor involving the two mechanisms of direct binding with gaseous H2S and suppressing the growth of VSC-producing oral bacteria

Enterococcus faecium WB2000 Inhibits Biofilm Formation by Oral Cariogenic Streptococci

This study investigated the inhibitory effect of probiotic Enterococcus faecium WB2000 on biofilm formation by cariogenic streptococci. The ability of E. faecium WB2000 and JCM5804 and Enterococcus faecalis JCM5803 to inhibit biofilm formation by seven laboratory oral streptococcal strains and 13 clinical mutans streptococcal strains was assayed. The Enterococcal strains inhibited biofilm formation in dual cultures with the mutans streptococcal strains Streptococcus mutans Xc and Streptococcus sobrinus JCM5176 (P < 0.05), but not with the noncariogenic streptococcal strains. Enterococcus faecium WB2000 inhibited biofilm formation by 90.0% (9/10) of the clinical S. mutans strains and 100% (3/3) of the clinical S. sobrinus strains. After culturing, the pH did not differ between single and dual cultures. The viable counts of floating mutans streptococci were lower in dual cultures with E. faecium WB2000 than in single cultures. Enterococcus faecium WB2000 acted as a probiotic bacterial inhibitor of cariogenic streptococcal biofilm formation

Application of a Chairside Anaerobic Culture Test for Endodontic Treatment

Periapical lesions are caused by bacterial infections. The fundamental objective of endodontic treatment is to eliminate bacteria present in the root canal system because they play an important role in the development and maintenance of periapical lesions. Therefore, confirming the absence of bacteria before filling root canals is important. Anaerobic culture tests have been used in many endodontic cases, and they have brought about good treatment outcomes. These tests, however, require specific apparatuses and bacteriological techniques. Here, we report a chairside anaerobic culture test that does not require any specialized apparatuses or techniques. We also report two endodontic cases in which this simple test was used. Both patients were diagnosed with chronic purulent periapical lesions. After confirming the absence of bacteria in the root canals, they were filled with gutta-percha points. At followup, the radiolucencies showed recovery, although longterm observation is under way. From these results, the authors conclude that this simple chairside anaerobic culture test is effective for evaluating periapical lesion treatment procedures

Effect of S-PRG Eluate on Biofilm Formation and Enzyme Activity of Oral Bacteria

Recently, the antibacterial activity of a composite resin containing prereacted glass ionomer (S-PRG) filler was revealed. We examined the effect of an S-PRG eluate on various biologic activities of Streptococcus mutans and Porphyromonas gingivalis. Adherence ability of S. mutans was evaluated by microtiter plate assay; protease and gelatinase activities of P. gingivalis were examined by synthetic substrate hydrolysis and gelatin film spot assay, respectively. Coaggregation of P. gingivalis with Fusobacterium nucleatum was also examined. S-PRG eluate was found to suppress streptococcal adherence. S-PRG eluate inhibited the protease and gelatinase activities of P. gingivalis and the coaggregation between P. gingivalis and F. nucleatum. These results indicate that S-PRG eluate suppresses streptococcal adherence and inhibits the protease and coaggregation activities of P. gingivalis. These findings may prompt research into novel strategies for preventing caries and periodontitis

Analisis Antibodi IgG Spesifik Fimbria Porphyromonas gingivalis dengan Metoda Elisa pada Penderita Early-Onset Periodontitis

The adherence of bacteria to the host tissues is a first step in the development of infections. Bacterial fimbriae have been shown to play an important role in the interaction between bacteria and host cells or among bacterial cells. Optical density of serum IgG specific Porphyromonas gingivalis strain ATCC 33277 fimbriae antibodies has been estimated in 32 cases of Early-Onset Periodontitis and 14 healthy periodontal subjects as control. The mean level of IgG were significantly elevated (p=0,000) in their cases. The data support the role of P. gingivalis as a key pathogen in Early-Onset Periodontitis