Identification of cell surface markers for acute myeloid leukemia prognosis based on multi-model analysis

Given the extremely high inter-patient heterogeneity of acute myeloid leukemia(AML),the identification of biomarkers for prognostic assessment and therapeutic guidance is critical.Cell surface markers(CSMs)have been shown to play an important role in AML leukemogenesis and progression.In the current study,we evaluated the prognostic potential of all human CSMs in 130 AML patients from The Cancer Genome Atlas(TCGA)based on differential gene expression analysis and univariable Cox proportional hazards regression analysis.By using multi-model analysis,including Adaptive LASSO regression,LASSO regression,and Elastic Net,we constructed a 9-CSMs prognostic model for risk stratification of the AML patients.The predictive value of the 9-CSMs risk score was further validated at the transcriptome and proteome levels.Multivariable Cox regression analysis showed that the risk score was an independent prognostic factor for the AML patients.The AML patients with high 9-CSMs risk scores had a shorter overall and event-free survival time than those with low scores.Notably,single-cell RNA-sequencing analysis indicated that patients with high 9-CSMs risk scores exhibited chemotherapy resistance.Furthermore,PI3K inhibitors were identified as potential treatments for these high-risk patients.In conclusion,we constructed a 9-CSMs prognostic model that served as an independent prognostic factor for the survival of AML patients and held the potential for guiding drug therapy.

Characterization of the osteogenic potential of mesenchymal stem cells from human periodontal ligament based on cell surface markers

Mesenchymal stem cell （MSC）-mediated therapy has been shown to be clinically effective in regenerating tissue defects. For improved regenerative therapy, it is critical to isolate homogenous populations of MSCs with high capacity to differentiate into appropriate tissues. The utilization of stem cell surface antigens provides a means to identify MSCs from various tissues. However, few surface markers that consistently isolate highly regenerative MSCs have been validated, making it challenging for routine clinical applications and making it all the more imperative to identify reliable surface markers. In this study, we used three surface marker combinations： CD51/CD140a, CD271, and STRO-1/CD146 for the isolation of homogenous populations of dental mesenchymal stem cells （DMSCs） from heterogeneous periodontal ligament cells （PDLCs）. Fluorescence-activated cell sorting analysis revealed that 24% of PDLCs were CD51＋/CD140a＋, 0.8% were CD271＋, and 2.4% were STRO-1＋/CD146＋. Sorted cell populations were further assessed for their multipotent properties by inducing osteogenic and chondrogenic differentiation. All three subsets of isolated DMSCs exhibited differentiation capacity into osteogenic and chondrogenic lineages but with varying degrees. CD271＋ DMSCs demonstrated the greatest osteogenic potential with strong induction of osteogenic markers such as DLX5, RUNX2, and BGLAP. Our study provides evidence that surface marker combinations used in this study are sufficient markers for the isolation of DMSCs from PDLCs. These results provide important insight into using specific surface markers for identifying homogenous populations of DMSCs for their improved utilization in regenerative medicine.

Effects of Nano-TiO2 on Chlamydomonas reinhardtii Cell Surface under UV,Natural Light Conditions

Cell surface of aquatic organisms constitutes a primary site for the interaction and a barrier for the nano-TiO2 biological effects.In the present study,the biological effects of nano-TiO2 on a unicellular green algae Chlamydomonas reinhardtii were studied by observing the changes of the cell surface morphology and functional groups under UV or natural light.By SEM,the cell surface morphology of C.reinhardtii was changed under UV light,nano-TiO2 with UV light or natural light,which indicated that photocatalysis damaged cell surface.It was also observed that cell surface was surrounded by TiO2 nanoparticles.The ATR-FTIR spectra showed that the peaks of functional groups such as C-N,-C=O,-C-O-C and P=O,which were the important components of cell wall and membrane,were all depressed by the photocatalysis of nano-TiO2 under UV light or natural light.The photocatalysis of nano-TiO2 promoted peroxidation of functional groups on the surface of C.reinhardtii cells,which led to the damages of cell wall and membrane.

Mechanobiology of the cell surface:Probing its remodeling dynamics using membrane tether pulling assays with optical tweezers

Mammalian cell surfaces consist of the plasma membrane supported by an underneath cortical cytoskeleton.Together,these structures can control not only the shape of cells but also a series of cellular functions ranging from migration and division to exocytosis,endocytosis and differentiation.Furthermore,the cell surface is capable of exerting and reacting to mechanical forces.Its viscoelastic properties,especially membrane tension and bending modulus,are fundamental parameters involved in these responses.This viewpoint summarizes our current knowledge on how to measure the viscoelastic properties of cell surfaces employing optical tweezers-based tether assays,paving the way for a better understanding of how cells react to external mechanical forces,with a glance on their remodeling dynamics and possible consequences on downstream cellular processes.

<i>In Vitro</i>Characterization of Cell Surface Properties of 14 Vaginal <i>Lactobacillus</i>Strains as Potential Probiotics

Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Human-origin Lactobacillus is a preferable source of probiotic bacteria. This study screened 14 vaginal Lactobacillus strains as probiotic candidates by investigating probiotic-related cell surface characteristics including cell surface hydrophobicity (CSH), Lewis acidity/basicity, autoaggregation, and biofilm formation. Moderate to high CSH and autoaggregation, high basicity and low acidity were prevalent in the 14 tested strains. Biofilm formation varied in a large range among the 14 tested strains. CSH showed a high correlation with Lewis acidity and autoaggregation, while Lewis acidity was highly correlated with autoaggregation and biofilm formation. Four strains were selected as promising probiotic strains. This study was the first one to compare antibiotic sensitivity between biofilm-forming cells and planktonic cells of Lactobacillus species, and found that biofilm-forming cells of a L. fermentum strain had a significantly higher survival rate than planktonic cells in cefotaxime, cefmetazole and tetracycline, but were as sensitive to oxacillin and ampicillin as planktonic cells were.

Aqueous Leaves Extract of Gongronema latifolium (Benth) Downregulates the Expression of IFN-γ, IL-10 and Cell Surface Markers in Rabbits

Background: The pathophysiology of the inflammatory process reveals intricate signaling which includes the IL-1β, IL-6, and TNFα pathways that could serve as drug targets. Aim: This study determined the effect of the aqueous extract of Gongronema latifolium (AEGL) leaves on the expression of IFNγ, IL-10, CD3, and CD56 in rabbits. Materials and Methods: ELISA tests were performed to determine the effect of the AEGL on the expression of a pro-inflammatory cytokine (IFNγ), an anti-inflammatory cytokine (IL-10), and CD3 and CD56 cell surface markers in rabbits. Twenty cross-bred male rabbits with an average weight range of 1.0 - 1.5 kg were selected. The rabbits were separated into four groups of four rabbits each treated as follows: Grp1 is the untreated control;Grp2 is the treated control;and Grp3, Grp4, and Grp5 were treated with 200, 400, and 600 mg/kg of AEGL respectively for 28 days. Results: The AEGL showed its greatest inhibitory effect in Group 4 on IL-10 (118.5 pg/ml), and IFNγ (332 pg/ml) on days 14 and 21 respectively. AEGL also showed the highest inhibition of CD3 expression on days 14 and 21 (0 pg/ml) in Group 3;and CD56 expression on day 21 (630.5 pg/ml) in Group 4. Conclusion: AEGL showed exhibited strong T cell mediated anti- inflammatory, and immunomodulatory activity in test rabbits within the 28-day period which can be confirmed by cell based assays. Specifically at 400 mg/kg, AEGL exhibited the greatest anti-inflammatory activity which is suggestive of its maximum effective dose.

Expression of phenylalanine ammonia lyase as an intracellularly free and extracellularly cell surface-immobilized enzyme on a gut microbe as a live biotherapeutic for phenylketonuria

Phenylketonuria(PKU),a disease resulting in the disability to degrade phenylalanine(Phe)is an inborn error with a 1 in 10,000 morbidity rate on average around the world which leads to neurotoxicity.As an potential alternative to a protein-restricted diet,oral intake of engineered probiotics degrading Phe inside the body is a promising treatment,currently at clinical stage II(Isabella,et al.,2018).However,limited transmembrane transport of Phe is a bottleneck to further improvement of the probiotic’s activity.Here,we achieved simultaneous degradation of Phe both intracellularly and extracellularly by expressing genes encoding the Phe-metabolizing enzyme phenylalanine ammonia lyase(PAL)as an intracellularly free and a cell surface-immobilized enzyme in Escherichia coli Nissle 1917(EcN)which overcomes the transportation problem.The metabolic engineering strategy was also combined with strengthening of Phe transportation,transportation of PAL-catalyzed trans-cinnamic acid and fixation of released ammonia.Administration of our final synthetic strain TYS8500 with PAL both displayed on the cell surface and expressed inside the cell to the Pah^(F263S)PKU mouse model reduced blood Phe concentration by 44.4%compared to the control Ec N,independent of dietary protein intake.TYS8500 shows great potential in future applications for PKU therapy.

A new photolabeling probe for efficient enrichment and deep profiling of cell surface membrane proteome by mass spectrometry

The cell surface membrane proteome is a class of proteins encoded by ~25% of all protein-coding genes in living organisms and plays a key role in mediating communication between the cells and their surrounding environment. However, most cell surface membrane proteins(CSMPs) are naturally expressed at very low levels compared with intracellular proteins. The difficulties in their purification with high specificity further hinder the understanding of their structure and function. In this study, we developed a new photolabeling probe to achieve efficient tagging and facile enrichment of the CSMPs. The probe is composed of a lipid tail for cell surface localization, a polyethylene glycol(PEG) spacer for increased water solubility, two 4-(N-maleimido)benzophenone(MBP) groups for UV-active tagging of the CSMPs, and a biotin tag for subsequent isolation. Application of this photolabeling probe resulted in the successful enrichment and identification of 3098 annotated CSMPs in HT22 cells with close to 70% selectivity. The proposed photolabeling probe and enrichment strategy were demonstrated to be a powerful method for deep cell surface proteome profiling, representing one of the largest groups of current drug targets.

Surface activity of cancer cells:The fusion of two cell aggregates

A key feature that distinguishes cancer cells from all other cells is their capability to spread throughout the body.Although how cancer cells collectively migrate by following molecular rules which influence the state of cell-cell adhesion contacts has been comprehensively formulated,the impact of physical interactions on cell spreading remains less understood.Cumulative effects of physical interactions exist as the interplay between various physical parameters such as(1)tissue surface tension,(2)viscoelasticity caused by collective cell migration,and(3)solid stress accumulated in the cell aggregate core region.This review aims to point out the role of these physical parameters in cancer cell spreading by considering and comparing the rearrangement of various mono-cultured cancer and epithelial model systems such as the fusion of two cell aggregates.While epithelial cells undergo volumetric cell rearrangement driven by the tissue surface tension,which directs cell movement from the surface to the core region of two-aggregate systems,cancer cells rather perform surface cell rearrangement.Cancer cells migrate toward the surface of the two-aggregate system driven by the solid stress while the surface tension is significantly reduced.The solid stress,accumulated in the core region of the two-aggregate system,is capable of suppressing the movement of epithelial cells that can undergo the jamming state transition;however,this stress enhances the movement of cancer cells.We have focused here on the multi-scale rheological modeling approaches that aimed at reproducing and understanding these biological systems.

Generation and characterisation of rabbit monoclonal antibodies against the native cell surface antigens of embryonic stem cells

Embryonic stem （ES） cells are potent resources for cell therapy,and monoclonal antibodies （mAbs） against native cell surface markers of ES cells could be useful tools for therapeutic applications.Here,we report the development of a feasible approach,which could be used in mass production,for experimentally producing rabbit mAbs against native cell surface antigens on the cell surface.Two of the 14 mAbs,which were selected at random,could be bound to the cell surface antigens of mES cells.The immunocytochemistry （ICC） and Western blot results showed that mAb 39 recognises conformational epitopes.The target antigen of mAb 39 was then successfully purified using an improved immunoprecipitation approach in which mAb was bounded to intact mES cells before the cells were lysed.The LC-LTQ mass spectrum analysis showed that the target antigen of mAb 39 was Glut3.This result was further confirmed by Western blot using commercially available antibodies against Glut3.Further experiments showed that mAb 39 exhibited an antiproliferative effect on mES cells.We also found that Glut3 was differentially expressed among the mES cell population as detected by flow cytometry.

Cell surface protein engineering for high-performance whole-cell catalysts

Cell surface protein engineering facilitated by accumulation of information on genome and protein structure involves heterologous production and modifica- tion of cell surface proteins using genetic engineering, and is important for the development of high-performance whole-cell catalysts. In this field, cell surface display is a major technology by exposing target proteins, such as enzymes, on the cell surface using a cartier protein. The target proteins are fused to the carrier proteins that transport and tether them to the cell surface, as well as to a secretion signal. This paper reviews cell surface display systems for prokaryotic and eukaryotic cells from the perspective of carrier proteins, which determine the number of displayed molecules, and the localization, size, and direction （N- or C-terminal anchoring） of the passengers. We also discuss advanced methods for displaying multiple enzymes and a new method for the immobilization of whole-cell catalysts using adhesive surface proteins.

The Plant Cell Surface

Multicellular organization and tissue construction has evolved along essentially different lines in plants and animals. Since plants do not run away, but are anchored in the soil, their tissues are more or less firm and stiff. This strength stems from the cell walls, which encase the fragile cytoplasm, and protect it. Properties of plant cell walls translate into properties of plant tissue. For instance, the cellulose microfibril angle in the different layers of walls of individual cells is a determinant of mechanical functions, which are useful to the plant itself. It also determines material properties of tissues and their potential industrial use. Indeed, plant cell walls determine the industrial value of a range of plant products including paper, timber, foodstuff, fodder, spun fibers, coatings, renewable polymers and future nanocomposites. Cell walls and their biosynthesis is a very active field of plant research.

Improvement of the Open Circuit Voltage of CZTSe Thin-Film Solar Cells by Surface Sulfurization Using SnS

The objective of this study is to find an effective method to improve Voc without Jsc loss for Cu2ZnSnSe4 （CZTSe） thin film solar cells, which have been fabricated by the one step co-evaporation technique. Surface sulfurization of CZTSe thin films is carried out by using one technique that does not utilize toxic H2S gas; a sequential evaporation of SnS after CZTSe deposition and the annealing of CZTSe thin films in selenium vapor. A Cu2ZnSn（S, Se）4 （CZTSSe） thin layer is grown on the surface of the CZTSe thin film after the annealing. The conversion efficiency of the completed device is improved due to the enhancement of Voc, which could be attributed to the formation of a hole-recombination barrier at the surface or the passivation of the surface and grain boundary by S incorporation.

The 18.3% Silicon Solar Cells with Nano-Structured Surface and Rear Emitter

A nano-structured surface is formed on the pyramid structure of n-type silicon solar cells by size-controlled silver nano-particle assisted etching. Such a nano-structure creates a front average weighted reflectance of less than 2.5% in the 300-1200nm range due to the broadband reflection suppression. The sodium hydroxide is used to obtain the low-area surface by post-etching the nano-structure, thus the severe carrier recombination associated with the nano-structured surface could be reduced. After emitter forming, screen printing and firing by means of the industrial fabrication protocol, an 18.3%-efficient nano-structured silicon solar cell with rear emitter is fabricated. The process of fabricating the solar cells matches well with industrial manufacture and shows promising prospects.

Inhibiting Smooth Muscle Cell Proliferation via Immobilization of Heparin/Fibronectin Complexes on Titanium Surfaces

The aim of this study was to investigate the inhibitory effect of heparin/fibronectin （Hep/Fn） complexes on neointimal hyperplasia following endovascular intervention. Hep/Fn complexes were immobilized onto titanium （Ti） surfaces, with subsequent X-ray photoelectron spectroscopy （XPS）, Toluidine Blue 0 （TBO） and immunohistochemistry methods were used to characterize surface properties. Smooth muscle cell （SMC） cultures were used to evaluate the effect of Hep/Fn complexes on SMC proliferation. Results showed that Hep/Fn complexes successfully immobilized onto Ti surfaces and resulted in an inhibition of SMC proliferation. This study suggests that Hep/Fn surface-immobilized biomaterials develop as a new generation of biomaterials to prevent neointimal hyperplasia, particularly for use in cardiovascular implants.

Identification,Characterization,and Probiotic Potentials of Lactobacillus pentosus SF-1

In recent years probiotics have been considered as a potential substitution of antibiotics to control pathogens and treat infectious diseases in aquaculture.In the present study a strain of Lactobacillus pentosus,named as L.pentosus SF-1,was isolated from waters in aquaculture.The species identification of this strain was conducted by 16S rRNA sequence,and the physiological and biochemical characteristics of this strain were assessed.Furthermore,the virulence,antibiotic sensitivity,cell surface characteristics and acid/base-resistance of L.pentosus SF-1 were determined to evaluate the probiotic potentials of this strain.Specifically,L.pentosus SF-1 is sensitive to most common antibiotics,and no hemolysin was generated from it,indicating the safety of this strain to hosts.In addition,L.pentosus SF-1 was able to tolerate the artificial gastric juice at pH 3 for 4 h and the artificial intestinal fluid at pH 6.8 or 8.0 for 6 h.Moreover,the analysis of self-aggregation and the adhesion of L.pentosus SF-1 to organic solvents suggested a high potential of L.pentosus SF-1 to inhabit the hosts,which was confirmed by testing the colonization of L.pentosus SF-1 in germ-free zebrafish.Interestingly,L.pentosus SF-1 displayed a high bactericidal activity against several bacterial pathogens.Consistently,the incubation of L.pentosus SF-1 significantly promoted the expression of antimicrobial components in zebrafish,contributing to the protection of the fish from E.tarda infection in vivo.Taken together,the probiotic strain L.pentosus SF-1 could be applied as anti-infection reagent in aquaculture.

Expression of CD44,CD24 and ESA in pancreatic adenocarcinoma cell lines varies with local microenvironment

BACKGROUND:Emerging evidence suggests that pancreatic adenocarcinoma is hierarchically organized and sustained by pancreatic cancer stem cells.Furthermore,elimination of these cells is possible and therapeutically relevant.This study aimed to investigate the expression patterns of pancreatic cancer stem cell surface markers CD44,CD24 and ESA in pancreatic adenocarcinoma cell lines and explore the influence of their local microenvironment.METHODS:Flow cytometry was used to analyze the expression patterns of CD44,CD24 and ESA in five pancreatic adenocarcinoma cell lines (PANC-1,PC-2,MIA-Paca-2,AsPC-1 and BxPC-3).In addition,the capacity for sphereformation in serum-free medium of four cell lines (PANC-1,PC-2,MIA-Paca-2 and BxPC-3) was assessed.Then,the same assays were performed when tumor cell spheres were developed.The role of sonic hedgehog (SHH) in cell spheres from PANC-1 and MIA-Paca-2 were also assessed by RT-PCR.RESULTS:CD44 and CD24 were detected in PANC-1.Only CD44 expression was detected in PC-2,MIA-Paca-2 and AsPC-1.CD44,CD24 and ESA were all detected in BxPC-3.Tumor cell spheres developed in PANC-1 and MIA-Paca-2 in serumfree medium.This was accompanied by an increase in CD24 expression and a decrease in CD44 expression in PANC-1.Interestingly,the expression of CD44 and CD24 returned to initial levels once the medium was changed back from serumfree to serum-containing medium.No significant change in the expression of CD44 was detected in MIA-Paca-2.Furthermore,the relative quantification of SHH mRNA in PANC-1 cell spheres was significantly higher than that in cells cultured in the serum-containing medium.CONCLUSION:The expression patterns of the pancreatic cancer stem cell surface markers CD44,CD24 and ESA were diverse in different pancreatic adenocarcinoma cell lines and changed with their local microenvironment.

Single CD271 marker isolates mesenchymal stem cells from human dental pulp

Mesenchymal stem cells （MSCs） are a promising tool in regenerative medicine due to their capacity to differentiate into multiple lineages. In addition to MSCs isolated from bone marrow （BMSCs）, adult MSCs are isolated from craniofacial tissues including dental pulp tissues （DPs） using various stem cell surface markers. However, there has been a lack of consensus on a set of surface makers that are reproducibly effective at isolating putative multipotent dental mesenchymal stem cel^s （~M^Cs）. II1 ~his stucly, we used clif^et（~nt combinations of surface markers （CD51/CD140a, CD271, and STRO-1/CD146） to isolate homogeneous populations of DMSCs from heterogeneous dental pulp cells （DPCs） obtained from DP and compared their capacity to undergo multilineage differentiation. Fluorescence-activated cell sorting revealed that 27.3% of DPCs were CD51＋/CD140a＋, 10.6% were CD271＋, and 0.3% were STRO-1＋/CD146＋. Under odontogenic conditions, all three subsets of isolated DMSCs exhibited differentiation capacity into odontogenic lineages. Among these isolated subsets of DMSCs, CD271＋ DMSCs demonstrated the greatest odontogenic potential. While all three combinations of surface markers in this study successfully isolated DMSCs from DPCs, the single CD271 marker presents the most effective stem cell surface marker for identification of DMSCs with high odontogenic potential. Isolated CD271＋ DMSCs could potentially be utilized for future clinical applications in dentistry and regenerative medicine.

Carbon material-based anodes in the microbial fuel cells

For the performance improvement of microbial fuel cells(MFCs),the anode becomes a breakthrough point due to its influence on bacterial attachment and extracellular electron transfer(EET).On other level,carbon materials possess the following features:low cost,rich natural abundance,good thermal and chemical stability,as well as tunable surface properties and spatial structure.Therefore,the development of carbon materials and carbon-based composites has flourished in the anode of MFCs during the past years.In this review,the major carbon materials used to decorate MFC anodes have been systematically summarized,based on the differences in composition and structure.Moreover,we have also outlined the carbon material-based hybrid biofilms and carbon material-modified exoelectrogens in MFCs,along with the discussion of known strategies and mechanisms to enhance the bacteria-hosting capabilities of carbon material-based anodes,EET efficiencies,and MFC performances.Finally,the main challenges coupled with some exploratory proposals are also expounded for providing some guidance on the future development of carbon material-based anodes in MFCs.