This aspect may have influenced the pattern of HR response observed in this study when isotonic solution was ingested. In the present study, no hydration also reduced global HRV after exercise. In relation to the SDNN (ms), despite presenting similar behavior in both conditions,

higher values were displayed in the hydrated condition. This finding confirms the influence check details of hydration on post-exercise cardiac autonomic stability. This study has some limitations. The minimum interval see more between the execution of control and experimental protocols was adhered to, however, some collections were completed over a period longer than a week, which may hinder the interpretation of the variables studied. Urine density was not determined at the end of the control protocol in this study, even though this might have

contributed to the consolidation 4EGI-1 chemical structure and interpretation of results. However, we were unable to collect urine from the subjects, as they were unable to urinate because they were not hydrated. Another important aspect refers to the use of supine rest and recovery conditions, considering that this exercise was performed in the upright position. Although we chose to compare rest and exercise in different positions, we believed Celecoxib that the modifications produced in the parameters during exercise were not influenced by the postural change. However, in addition to being more tolerable for the volunteer, the choice of the supine position during the recovery period has not impaired the results since the parameters were compared to a baseline, with subjects in the same position. Considering the importance of the issue presented, other studies are in progress to evaluate the influence of water intake on cardiac autonomic

modulation and cardiorespiratory parameters. Water ingestion provides rapid gastric emptying, requires no adaptation to the palatability of the solution and offers an economic alternative [39], aspects that are important in the context of hydration during and after exercise. These studies will allow us to evaluate the influence of water intake as a rehydration drink and to compare the effects of the ingestion of isotonic solutions and water as a means of rehydration on cardiac autonomic modulation. Such studies may enrich the knowledge in exercise physiology. Conclusions We concluded that regardless of hydration status, the exercise protocol caused alterations in cardiac autonomic modulation, characterized by increased sympathetic and decreased parasympathetic activity.

These data suggest that survivin plays an important role in prompting the development of lung cancer. In recent years, studies have showed that the activity of survivin BMS202 datasheet promoter in tumor cells is significantly increased [24–27]. This suggests that the expression of survivin is transcriptional regulated. Reduction of promoter activity could significantly decrease the mRNA and thus decrease the protein expression of survivin. Although the survivin promoter contains several GC boxes,

but methylation of these GC boxes has not been found in the survivin promoter. It is implicit that the regulation of survivin expression is at the level of transcription but it is still unclear how survivin transcription is regulated by the Cis-acting elements. HIF-1α is highly expressed in various tumor tissues and plays an important Poziotinib cost role in regulating hypoxia, and tumor invasion and progress [17, 19, 20]. In this study, we confirmed that HIF-1α is highly expressed in NSCLC tissue, as was found in breast cancer [28]. The expression of HIF-1α is related to differentiation,

lymph node metastasis and clinical stage of lung cancer. Correlation analysis showed the expression of survivin was positively correlated with HIF-1α. The previous studies have showed that HIF-1α is intermediate link in the evolution of the tumor, and this protein could regulate a variety of hypoxia-induced gene expression [29]. In vitro, we also found that the expressions of HIF-1α and survivin AZD3965 in vivo in A549 cells were significantly increased under hypoxic conditions. Therefore, we speculated that HIF-1α might be a transcriptional activator of survivin. An early study using bioinformatic analysis of the survivin promoter 5′-upstream MRIP non-coding

region found that the survivin gene TSS (transcriptional start site) was located in -64 bp upstream of translation initiation codon (ATG). This bioinformatic analysis also showed that the potential transcription factors that could bind to the survivin promoter included Sp1, E2F, p53, CDE, CHR, etc [14]. Our study detected that there are also 2 putative binding sites for HIF-1α, which are located at-16 bp to -19 bp and at -133 bp to -136 bp in the proximal promoter region of human survivin gene. The first site (16 bp to -19 bp) partially overlaps with one of the potential Sp1 binding sites. Peng et al [20] also confirmed that there is a putative HIF-1α binding site in the survivin core promoter (-203 to +27). They also found that in breast cancer cells, HIF-1α, induced by EGF, could bind to this putative binding-site under hypoxic or normoxic conditions and that when HIF-1α is bound to its binding site in the survivin promoter the expression of survivin is increased [20].

PubMedCrossRef 61. Azad AK, Sadee W, Schlesinger LS: Innate immune gene polymorphisms in tuberculosis. Infect Immun 2012,80(10):3343–3359.PubMedCrossRef 62. Herrmann

JL, O’Gaora P, Gallagher A, Thole JE, Young DB: Bacterial glycoproteins: BAY 80-6946 ic50 a link between glycosylation and proteolytic cleavage of a 19 kDa antigen from Mycobacterium tuberculosis. Embo J 1996,15(14):3547–3554.PubMed 63. Tjalsma H, van Dijl JM: Proteomics-based consensus prediction of protein retention in a bacterial membrane. Proteomics 2005,5(17):4472–4482.PubMedCrossRef 64. Zhang YJ, Ioerger TR, Huttenhower C, Long JE, Sassetti CM, Sacchettini JC, Rubin EJ: Global assessment of genomic regions required for growth in Mycobacterium tuberculosis. PLoS Pathog 2012,8(9):e1002946.PubMedCrossRef 65. Robichon C, Vidal-Ingigliardi D, Pugsley AP: Depletion of apolipoprotein N-acyltransferase selleck compound causes mislocalization of outer membrane lipoproteins in Escherichia coli. J Biol Chem

2005,280(2):974–983.PubMedCrossRef 66. Niederweis M, Danilchanka O, Huff J, Hoffmann C, Engelhardt H: Mycobacterial outer membranes: in search of proteins. Trends Microbiol 2010,18(3):109–116.PubMedCrossRef 67. Sutcliffe IC: A phylum level perspective on bacterial cell envelope architecture. Trends Microbiol 2010,18(10):464–470.PubMedCrossRef 68. Zuber B, Chami M, Houssin C, Dubochet J, Griffiths G, Daffe M: Direct visualization of the outer membrane of mycobacteria and corynebacteria in their native state. J Bacteriol 2008,190(16):5672–5680.PubMedCrossRef GNAT2 Competing interests The authors declare that they have no competing interests. Authors’ contributions JKB designed the study, performed experimental work and drafted the manuscript. AT carried out the genetic engineering of M. bovis BCG mutant strain and participated in the MS/MS data analyses. PS conceived of the study,

participated in its coordination and helped to draft the manuscript. All authors read and approved the final manuscript.”
“Background Coxiella burnetii is a highly infectious Gram-negative intracellular bacterium that causes the zoonosis Q fever [1]. Central to C. burnetii ��-Nicotinamide cost pathogenesis is the ability to proliferate within a parasitophorous vacuole (PV) of macrophages that has characteristics of a large phagolysosome [2, 3]. By unknown mechanisms, the pathogen can resist the degradative activities of the vacuole while exploiting the biochemical and biophysical properties of the PV to promote robust intracellular replication [4, 5]. The C. burnetii PV is a unique cellular compartment that can occupy nearly the entire host cell cytoplasm [6]. C. burnetii protein synthesis is required for PV interactions with a subset of cellular vesicles that contribute material to the growing vacuole [7, 8].

To further confirm that single point mutation only causing SHP099 non-synonymous mutation was truly involved in the serotype shift, we performed the opposite experiment, i. e. induction of Inaba serotype in Ogawa strains. We created the T472C substitution on the chromosomal rfbT gene of Ogawa strain 7743 through homologous recombination. As expected, this substitution caused serotype shift from Ogawa to Inaba in strain 7743. Subsequent introducing the recombinant plasmid pBR-rfbT carrying intact the rfbT gene induced the seroconversion from Inaba to Ogawa phenotype. Taken together, our study experimentally demonstrated T472C substitution is truly involved in the serotype shift. Discussion In this study,

we presented the descriptive data regarding cholera serotype-cycling in China over a 48-year (1961–2008) period, GDC-0449 research buy and also noted the multiplicity of rfbT sequence variations in V. cholerae O1 isolates. Three single nucleotide substitutions and deletion mutations of rfbT have been reported which caused serotype switching due to a frameshift or crucial amino acid residue change in RfbT [22, 41, 42]. In our study much more mutations are found in the Inaba

serotype strains, including single amino acid residue substitutions, frameshifts caused by single nucleotide and short fragment insertions/deletions, and transposition events. These mutations occurred randomly over the entire open reading frame of rfbT, which may suggest the mutations occurred frequently and differently under pressures from environment and human immunity, as well as spontaneous mutation. With the complementation of the intact rfbT gene, these Inaba strains were

converted to the Ogawa serotype, which validate the mutations on the Inaba serotype conversion. Our study provides the first evidence that mobile genetic elements, including the transposase OrfAB and ISVch5 transposase, are involved in inactivating the rfbT of V. cholerae, thus contributing to PD184352 (CI-1040) the serotype interconversion. The insertion of the two kinds of transposases both led to duplication of the inserted sequence. Although there is difference in terms of the insertion position and orientation, the target sequence (AAAC) of the transposase OrfAB elements in different strains was the same. We further surveyed the distribution of transposase OrfAB copies in click here several strains which genome sequences are available. The copy number and the distribution of transposase OrfAB on chromosomes I and/or II vary in strains from different regions and years. Strain N16961 contains six copies, each chromosome harbors three copies. In IEC224, in addition to the three copies on each chromosome, there is an additional transposase OrfAB subunit B on chromosome I. In strain MJ-1236, all four copies are located on chromosome II. All these and our data suggest that the transposase OrfAB is quite active in transposition in V.

6 to 4.1 nm), and the globular structure appears on the glass. Further increase of Au thickness leads to the increase of layer’s homogeneity and the globular structure being less pronounced as well as the surface roughness. The thermal annealing Autophagy inhibitor screening library leads to a significant increase of surface roughness (Figure 3, second column). The globular structure is strongly amplified probably due to the local surface melting of gold nanoparticles during the thermal annealing process [16]. The dimensions of globular structures

are significantly higher in comparison to non-annealed ones. The surface morphology of the annealed Au with thickness of 35 nm is similar to those observed on glass substrate deposited by sputtering [15]. Similar changes in the morphology of the thin gold annealed structures and a sharp increase in surface roughness were observed on the samples annealed at 200°C for 20 h [17] and at 450°C for 2 h [18]. Figure 3 AFM images of the evaporated Au layers at different temperatures. AFM images of the evaporated Au layers on glass with room temperature

(first column, RT) and the same samples consequently annealed at 300°C (second column, annealed). The thicknesses of evaporated Au were 7, 18, and 35 nm. R a is the arithmetic mean surface roughness in nanometers. The rather different appearance of surface morphology was determined for evaporated Au deposited on check details glass already heated to 300°C (Figure 4). The gold layer of 7-nm thickness exhibited globular nanostructure with roughness of 3.8 nm. With increasing Au layer thickness, the globular nanostructure has a tendency to disappear. The electrically continuous nanolayer (35 nm) exhibits the lowest values of surface roughness (1.7 nm), the surface Oxymatrine pattern being similar to those obtained for sputtered Au [19]. The reason for such appearance should be within the formation of nanolayer and its nucleation. The electrical measurement revealed that the difference in thickness when the electrically continuous layer (Figure 1) is formed for buy LY2603618 as-evaporated and consequently

annealed layer and is minor in comparison to previously studied annealing of sputtered Au [5]. Therefore, we can suppose that the surface diffusion of gold nanoparticles is suppressed when the layer is heated, which is connected with the different surface wettability when the substrate is heated. The influence of surface diffusion may take place also in the case of evaporation in the already heated glass (Figure 4). The appearance of globular structures caused by the evaporation of 7-nm Au is probably caused by the surface melting of evaporated Au nanoparticles during the deposition process. Even when the melting process takes place, the surface diffusion is suppressed and the structure has regular and homogeneous character.

Four weeks after initial treatment, all mice were sacrificed to assess the effects of drug treatments. All procedures involving mice complied with the Guide for the Care and Use of Laboratory Animals (National Institutes of Health). Western blotting The tissues were homogenized in 0.5 ml Hepes (50 mM, pH 7.5) containing 100 mM NaCl, 1 mM CaCl2, 1 mM dithiothreitol,

1% ethylene find more glycol-bis(aminoethyl ether)-tetraacetic acid 1% Triton X- 100 and proteinase inhibitors. Protein extracts were kept in ice for 30 min and then centrifuged at 14,000 PARP inhibition g at 4°C for 30 min. Protein concentrations were determined using a bicinchoninic acid protein assay reagent kit. Protein samples (20 mg) were mixed with equal volumes of loading buffer (20% glycerol, 4% check details sodium dodecyl sulfate, and 100 mM Tris-HCl, pH 6.8) and then boiled for 5 min in the presence of β-mercaptoethanol. Proteins were separated in 8% sodium dodecyl sulfate-polyacrylamide gels at 100 V for 2 h and then electrotransferred to nitrocellulose membranes at 270 mA for 2 h. Membranes were blocked with 5% non-fat dry milk in PBS with 0.1% Tween 20 for 1 h at room temperature. Then, membranes were incubated with anti- HIF-1α (1:500) overnight at 4°C

and finally with a horseradish peroxidase-conjugated anti-mouse IgG for 1 h at room temperature after washing with TBS containing 0.1% Tween 20. Proteins were visualized by enhanced chemiluminescence reagents after washing. Protein expression was semi-quantified using an image analysis system. CD34-PAS dual staining Four micrometer paraffin sections were routinely deparaffinized and dehydrated. First, CD34 immunohistochemical staining was applied to the sections. Endogenous peroxidase activity was blocked with

3% hydrogen peroxide in 50% methanol for 10 min at room temperature. Sections were rehydrated and washed with PBS and then pretreated with citrate buffer (0.01 M citric acid, pH 6.0) for 20 Docetaxel min at 100°C in a microwave oven. Non-specific binding sites were blocked with 2% normal goat serum in PBS for 20 min at 37°C. Sections were then incubated overnight at 4°C with anti-CD34 at a 1:200 dilution. Then, sections were rinsed with PBS and incubated with biotinylated goat anti-mouse IgG for 20 min at 37°C, followed by incubation with 3,3′-diaminobenzidine(DAB) chromogen for 10 min at room temperature. Sections were then rinsed with water for 1 min to stop the DAB-staining reaction. Formalin and melanin granules were then removed using the methods mentioned above. Finally, sections were treated with 0.5% periodic acid solution for 10 min and rinsed with distilled water for 2-3 min. In a dark chamber, sections were treated with Schiff solution for 15-30 min. After rinsing with distilled water, sections were counterstained with hematoxylin [9].

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) are found in ~50% of all bacterial species, including Salmonella[27]. CRISPR elements comprise several unique short sequences, called spacers, which are interspaced

PND-1186 cost by conserved direct repeats. In some bacteria, homology between a spacer and a complementary target nucleic acid results in degradation of the target by sequence-specific endonucleases, providing protection from exogenous bacteriophage or plasmid DNA [reviewed in [28]. Due to both acquisition and loss of these spacer elements, CRISPRs represent arguably the most rapidly evolving prokaryotic loci [29–31]. Sequence analysis of CRISPR loci has been used to subtype clinical isolates of Salmonella[32–34], Escherichia coli[35, 36], group A Streptococcus[37] and Campylobacter species [38]. Salmonella contains two of these non-coding loci, which are comprised of direct selleck repeats of 29 nucleotides separated by spacers of 32 nucleotides (Figure 1). Generally, CRISPR polymorphisms between Salmonella strains are due to deletion or repetition of one or more spacers, termed ‘spacer microevolution’ [32–34, 39, 40]. An extensive investigation of 738 isolates, representing several different serovars, showed that polymorphisms within

the CRISPR loci correlate highly with serovar, with isolates from individual serovars bearing distinct CRISPR patterns [32]. Figure 1 Salmonella CRISPR loci. Salmonella have two CRISPR loci, selleck inhibitor CRISPR1 and CRISPR2 comprised of direct repeats of 29 nucleotides (black diamonds) separated by spacers (empty rectangles). There is an A-T rich leader sequence why upstream of each locus (shaded rectangle) and the CRISPR-associated genes (cas) are upstream of the CRISPR1

locus (grey boxed arrow). Primers used for amplification are shown in blue and red for CRISPR1 and CRISPR2, respectively. We recently developed a sequence-based subtyping assay (multi-virulence locus sequence typing; MVLST) for Salmonella that involves the sequencing of two virulence genes, fimH1 (fimH) and sseL, in addition to CRISPR sequencing [33]. Preliminary studies showed that this approach, termed CRISPR-MVLST, provided better discrimination than either CRISPR or MVLST alone and, importantly, exhibited strong epidemiologic concordance among eight out of nine of the most common illness-causing Salmonella enterica serovars [33], including both S. Heidelberg and S. Typhimurium outbreak strains. Subsequently, among a large number of clinical isolates of the highly clonal S. Enteritidis, a combination of CRISPR-MVLST and PFGE was required to provide a sufficient discriminatory power [34]. Among a large set of S. Newport clinical isolates, CRISPR-MVLST provides similar discrimination to PFGE [41]. To further determine the functionality of this new subtyping approach, we investigated the discriminatory power of both CRISPR-MVLST and PFGE among a larger and unbiased collection of clinical S. Typhimurium and S.

PubMedCrossRef 10. Sánchez B, Bressolier P, Urdaci MC: Exported proteins in probiotic bacteria: adhesion to intestinal surfaces, host immunomodulation and molecular cross-talking with the host. FEMS Immunol Med Microbiol 2008, 54:1–17.PubMedCrossRef 11. Tjalsma H, Lambooy L, Hermans PW, Swinkels DW: Shedding & shaving: disclosure of proteomic expression on a bacterial face.

Proteomics 2008, 8:1415–1428.PubMedCrossRef 12. Munoz-Provencio D, Perez-Martinez G, Monedero V: Identification RG7112 mouse of surface proteins from Lactobacillus casei BL23 able to bind fibronectin and collagen. Probiotics & Antimicro Prot 2011, 3:15–20.CrossRef 13. Esko J, Lindahl U: Molecular diversity of heparan sulfate. J Clin Invest 2001, 108:169–173.PubMed 14. Prydz K, Dalen KT: Synthesis and sorting of proteoglycans. J Cell Sci 2000, 113:193–205.PubMed 15. Turnbull J, Powell A, Guimond S: Heparan sulpatem decoding a dynamicl multifunctional cell regulator. TRENDS Cell Biol 2001, 11:75–82.PubMedCrossRef 16. Bernfield M, Götte M, Park PW, Reizes O, Fitzgerald M, Lincecum J, Zako M: Functions of cell surface heparan sulfate proteoglycans. Annu Rev Biochem 1999, 68:729–777.PubMedCrossRef 17. Rapraeger A, Jalkanen M, Bernfield M: Cell

surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells. J Cell Biol 1986, 103:2683–2696.PubMedCrossRef 18. Schmidt G, Robenek H, Harrach B, Glössl J, Nolte V, Hörmann H, Richter H, Kresse H: Interaction of small dermatan sulfate proteoglycan from fibroblasts with

fibronectin. J Cell Biol 1987, 104:1683–1691.PubMedCrossRef SCH727965 19. Kirn-Safran C, Farach-Carson MC, Carson DD: Multifunctionality of extracellular and cell surface heparan sulfate proteoglycans. Cell Mol Life Sci 2009, 66:3421–3434.PubMedCrossRef 20. Schaefer L, Schaefer RM: Proteoglycans: from structural Sitaxentan compounds to signaling molecules. Cell Tissue Res 2010, 339:237–246.PubMedCrossRef 21. de Vries FP, Cole R, Dankert J, Frosch M, van Putten JP: check details Neisseria meningitidis producing the Opc adhesin binds epithelial cell proteoglycan receptors. Mol Microbiol 1998, 27:1203–1212.PubMedCrossRef 22. Chen T, Belland RJ, Wilson J, Swanson J: Adherence of pilus- Opa+ gonococci to epithelial cells in vitro involves heparan sulfate. J Exp Med 1995, 182:511–517.PubMedCrossRef 23. Grant CC, Bos MP, Belland RJ: Proteoglycan receptor binding by Neisseria gonorrhoeae MS11 is determined by the HV-1 region of OpaA. Mol Microbiol 1999, 32:233–242.PubMedCrossRef 24. Dupres V, Verbelen C, Raze D, Lafont F, Dufrêne YF: Force spectroscopy of the interaction between mycobacterial adhesins and heparan sulphate proteoglycan receptors. Chemphyschem 2009, 10:1672–1675.PubMedCrossRef 25. Sava IG, Zhang F, Toma I, Theilacker C, Li B, Baumert TF, Holst O, Linhardt RJ, Huebner J: Novel interactions of glycosaminoglycans and bacterial glycolipids mediate binding of enterococci to human cells. J Biol Chem 2009, 284:18194–18201.PubMedCrossRef 26.

Gut microbiota is acquired during early life and intestinal colonization starts immediately after birth. The ability of species to establish themselves durably in the colonic ecosystem depends on complex interactions between host and bacteria as well as between the bacteria themselves [3]. A wide range of factors may influence the establishment of the intestinal microbiota, including

type of delivery, feeding pattern, antibiotic therapy, contact with parents, siblings and hospital staff [4]. The nature of the gut flora, colonic bacterial metabolic pathways, the partial HDAC activity assay pressure of hydrogen in the colon, the buffering capacity of the colon, and incomplete monosaccharide absorption may play a part in infantile colic. Miller reported an increased breath hydrogen excretion in subjects suffering from infantile colic [5]. In 1994, Lehtonen et al. observed that an inadequate lactobacilli level occurring in the first months of life may affect the intestinal

HSP990 fatty acids profile and could favour the NU7026 nmr development of infantile colic [6]. Treem suggested that colicky infants produce large amounts of gas probably by colonic bacterial fermentation of malabsorbed dietary carbohydrate and that they are relieved of symptoms by the passage of gas [7]. It has also been demonstrated that less methane is produced by stool of colicky infants and this could be due to an inability of the gut microbiota to convert hydrogen to methane with a gastrointestinal hydrogen accumulation [8].

Moreover few old data support the notion that colicky infants produce more breath hydrogen in the fasting state and in response to feedings, which is thought to be evidence of lactose intolerance [9–11]. Differences in gut microbiota have been found Tenoxicam among colicky and non-colicky infants: colicky infants are less frequently colonized by Lactobacillus spp. and more frequently by anaerobic gram-negative bacteria [12]. Further, different colonization patterns of lactobacilli have been found among colicky and healthy infants: L. brevis and L. lactis are present only in colicky infants while L. acidophilus was detected only in healthy ones [13]. The recent finding that L. reuteri improve colic symptoms in breastfed infants suggested that a peculiar composition of the intestinal microbiota could favour the development of such disturbance [14, 15]; however the mechanisms through which lactic acid bacteria act on colic symptoms remain speculative.

The ratio imaging was conducted on fluorescent microscope

(Olympus, IX71-32PH, Shinjuku-ku, Tokyo, Japan). The PLGA microsphere was excited at 335 and 381 nm, and the images emitted at 452 and 521 nm were taken for analysis. The fluorescent intensity was analyzed using the software, WASABI V.1.4. The standard curve of ratio of fluorescent intensity vs. pH was generated by placing the LysoSensor™ Yellow/Blue dextran-loaded dextran nanoparticles at a known pH on a microscope slide. Multiple images were taken at each pH and then averaged to obtain the standard curve. Results and discussion Morphology of dextran nanoparticle The strategy for fabricating dextran nanoparticles loaded with proteins is shown in Figure 1. Briefly, proteins and PEG were dissolved in dextran solutions and aqueous solution, respectively. After these two solutions were mixed click here to get a clear solution, the solution was frozen dried under vacuum and washed with dichloromethane selleck products to give fine dextran nanoparticles loaded with proteins. Figure 1 The BMS202 concentration formulation strategy of fabricating the dextran nanoparticles loaded with proteins. Figure 2 shows SEM images of dextran nanoparticles loaded with BSA (DP-BSA).

DP-BSA exhibit a spherical shape, smooth surfaces, and diameters ranging from 200 to 500 nm. These results are consistent with that of the particle size analysis which shows the effective diameter of 293 nm for DP-BSA (Figure 3). Figure 2 An SEM photo of dextran nanoparticles loaded with BSA. Figure 3 The size distribution of dextran nanoparticles

loaded with BSA. Encapsulation efficiency of dextran nanoparticles As shown in Table 1, the encapsulation efficiency of dextran nanoparticles loaded with different proteins was generally larger than 98%. The recovery of proteins extracted from dextran nanoparticles ranged from 65% to 72%. Some proteins might be washed away by dichloromethane during the preparation Resminostat process. Table 1 The encapsulation efficiency and recovery of dextran nanoparticles ( n = 3) Number Protein Encapsulation efficiency(ave% ± SD) Recovery (%) (ave% ± SD) 1 BSA 99.23 ± 1.69 71.26 ± 2.06 2 GM-CSF 98.37 ± 1.27 69.16 ± 2.78 3 MYO 98.16 ± 1.55 65.57 ± 1.56 Protein aggregation during the formulation steps In order to address this novel dextran nanoparticle that may protect proteins from aggregation during the formulation process, the BSA, GM-CSF, and G-CSF were selected as model proteins, and SEC-HPLC was used to characterize the protein extracted from the protein standard solution, dextran nanoparticle, and controlled W/O emulsion. Figure 4 shows the SEC-HPLC charts of BSA extracted from the BSA standard solution, dextran nanoparticle, and W/O emulsion. The peak of BSA samples around 9.8 and 8.2 min were ascribed to the monomer and dimer BSAs, respectively. As shown in Figure 4, only one peak corresponding to the monomer BSA was observed in the BSA solution and dextran nanoparticle.