Dendrobium officinale polysaccharides-chemical properties and pharmacodynamic effects on the airways in experimental conditions.

The study aimed to analyze the effects of Dendrobium polysaccharides on the cough and airway reactivity and compare them with the effects of clinically used antitussives (codeine phosphate and butamirate citrate) and bronchodilators (salbutamol), using the guinea pig test system. Dendrobium officinale polysaccharides contained proteins (4.0 wt%) and phenolic compounds (1.7 wt%) with a molecular weight of 25,000 g/mol. The sugar analysis revealed a dominance of glucose (93.7 wt%) and a lesser amount of mannose (5.1 wt%) while other sugar quantities were negligible. Methylation analysis indicated the presence of highly branched polysaccharides. Glucose was found mainly as terminal, 1,4- and 1,6-linked. Furthermore, some 1,4- and 1,6-linked glucose units were found branched at O2, O3, and O6/O4. Mannose was terminal and 1,4-linked. NMR spectra signals indicate the presence of the (1→4)-linked α-d-glucan, (1→4)-linked ß-d-glucan branched at position O6, (1→6)-linked ß-d-glucan branched at position O3 and (1→4)-linked glucomannan. Pharmacological studies showed statistically significant antitussive activity of Dendrobium polysaccharides, exceeding the effect of clinically used antitussives, which may be partially associated with confirmed bronchodilation and the ability of polysaccharides to increase the threshold of cough receptor activation. Dendrobium polysaccharides may increase the possibility of symptomatic treatment of cough, especially in asthmatics.

Pineapple Lectin AcmJRL Binds SARS-CoV-2 Spike Protein in a Carbohydrate-Dependent Fashion.

The highly glycosylated spike protein of SARS-CoV-2 is essential for infection and constitutes a prime target for antiviral agents and vaccines. The pineapple-derived jacalin-related lectin AcmJRL is present in the medication bromelain in significant quantities and has previously been described to bind mannosides. Here, we performed a large ligand screening of AcmJRL by glycan array analysis, quantified the interaction with carbohydrates and validated high-mannose glycans as preferred ligands. Because the SARS-CoV-2 spike protein was previously reported to carry a high proportion of high-mannose N-glycans, we tested the binding of AcmJRL to the recombinantly produced extraviral domain of spike protein. We could demonstrate that AcmJRL binds the spike protein with a low-micromolar KD in a carbohydrate-dependent fashion.

Mannose-Anchored Nano-Selenium Loaded Nanostructured Lipid Carriers of Etravirine for Delivery to HIV Reservoirs.

The present investigation aims to develop and explore mannosylated lipid-based carriers to deliver an anti-HIV drug, Etravirine (TMC) and Selenium nanoparticles (SeNPs), to the HIV reservoirs via the mannose receptor. The successful mannosylation was evaluated by the change in zeta potential and lectin binding assay using fluorescence microscopy. Electron microscopy and scattering studies were employed to study the structure and surface of the nanocarrier system. The presence of selenium at the core-shell of the nanocarrier system was confirmed by X-ray photoelectron spectroscopy and energy dispersive X-ray analysis. Further, the in vitro anti-HIV1 efficacy was assessed using HIV1 infected TZM-bl cells followed by in vivo biodistribution studies to evaluate distribution to various reservoirs of HIV. The results exhibited higher effectiveness and a significant increase in the therapeutic index as against the plain drug. The confocal microscopy and flow cytometry studies exhibited the efficient uptake of the coumarin-6 tagged respective formulations. The protective effect of nano selenium toward oxidative stress was evaluated in rats, demonstrating the potential of the lipidic nanoparticle-containing selenium in mitigating oxidative stress in all the major organs. The in vivo biodistribution assessment in rats showed a 12.44, 8.05 and 9.83-fold improvement in the brain, ovary, and lymph node biodistribution, respectively as compared with plain TMC. Delivery of such a combination via mannosylated nanostructured lipid carriers could be an efficient approach for delivering drugs to reservoirs of HIV while simultaneously reducing the oxidative stress induced by such long-term therapies by co-loading Nano-Selenium.

Structural characterization of a pure polysaccharide from Bletilla striata tubers and its protective effect against H2O2-induced injury fibroblast cells.

The present study investigated the structural characteristics and its protective effect against H2O2-induced injury fibroblast cells of Bletilla striata tuber polysaccharide. The polysaccharides were gently extracted by water and recovered using the method of alcohol precipitation, and after further purification by DEAE-Sepharose Fast Flow gel column, a pure polysaccharide (pBSP) was finally obtained. The structural characterization of pBSP were investigated by using periodate oxidation studies, Smith-degradation, FT-IR spectroscopy, 1D and 2D NMR spectroscopy. The antioxidant effect of pBSP was evaluated by inhibiting the production of reactive oxygen species (ROS) in human fibroblast model cells induced by H2O2. It was firstly reported that pBSP was composed of d-glucose and D-mannose in a molar ratio of 1.00:1.34 with a molecular weight of 327.6 kDa. The repeating units of pBSP contained (1 â†’ 4)-linked-ß-D-Manp, (1 â†’ 4)-linked-α-D-Glcp and (1 â†’ 3)-linked-ß-D-Manp, and there was no branched chain. pBSP exhibited no toxic effect on fibroblasts cells and could protect them against H2O2-induced injuries. After pretreatment with pBSP for 24 h, the content of ROS in fibroblasts decreased significantly. These results not only confirm the availability B. striata, but also indicate that pBSP have potential antioxidant capacity. Our observations can provide foundation for further development of pBSP-based cosmetics.

Structural investigation of a thermostable 1,2-ß-mannobiose phosphorylase from Thermoanaerobacter sp. X-514.

1,2-ß-Mannobiose phosphorylases (1,2-ß-MBPs) from glycoside hydrolase 130 (GH130) family are important bio-catalysts in glycochemistry applications owing to their ability in synthesizing oligomannans. Here, we report the crystal structure of a thermostable 1,2-ß-MBP from Thermoanaerobacter sp. X-514 termed Teth514_1789 to reveal the molecular basis of its higher thermostability and mechanism of action. We also solved the enzyme complexes of mannose, mannose-1-phosphate (M1P) and 1,4-ß-mannobiose to manifest the enzyme-substrate interaction networks of three main subsites. Notably, a Zn ion that should be derived from crystallization buffer was found in the active site and coordinates the phosphate moiety of M1P. Nonetheless, this Zn-coordination should reflect an inhibitory status as supplementing Zn severely impairs the enzyme activity. These results indicate that the effects of metal ions should be taken into consideration when applying Teth514_1789 and other related enzymes. Based on the structure, a reliable model of Teth514_1788 that shares 61.7% sequence identity to Teth514_1789 but displays a different substrate preference was built. Analyzing the structural features of these two closely related enzymes, we hypothesized that the length of a loop fragment that covers the entrance of the catalytic center might regulate the substrate selectivity. In conclusion, these information provide in-depth understanding of GH130 1,2-ß-MBPs and should serve as an important guidance for enzyme engineering for further applications.

Comparative study of structural properties and biological activities of polysaccharides extracted from Chroogomphus rutilus by four different approaches.

Extraction processes significantly alter the structural and functional properties of polysaccharides. In this study, we extracted polysaccharides from Chroogomphis rutilus fruiting bodies (designated as CRP) using four methods, including hot water, ultrasound, microwave and sequential ultrasound-microwave, and designated these polysaccharides as CRP-H, CRP-M, CRP-U and CRP-UM, respectively. All CRPs were heteropolysaccharides with semblable monosaccharide types of glucose, mannose and galactose, mainly constituted of α-d-glucopyranosyl-(1 â†’ 4). The extraction processes significantly affected the molecular weights, monosaccharide proportions, glycosidic bond ratios, branching degrees, triple-helix conformation and surface morphology of the CRPs. Among them, CRP-UM showed the highest yield and most potent antioxidative capacity in vitro and in HL-7702 cells, but the weakest activation of immunostimulatory response in RAW264.7 cells. In contrast, CRP-H exhibited the lowest yield but strongest immunostimulatory activity. Overall, microwave extraction could be utilized as a general and practical CRP extraction approach, based on its relatively high yield and bioactivities.

Study on a novel spherical polysaccharide from Fructus Mori with good antioxidant activity.

A novel polysaccharide (MFP1P) was isolated from Fructus Mori, followed by purification via DEAE-52 cellulose and 27 % ethanol fraction. The MFP1P had the molecular weight of 56.78 kDa and the total sugar content of 93.32±0.54 %. And the MFP1P is mainly composed of glucose, galactose, galacturonic acid and mannose with molar ratio of 66.62 %, 13.94 %, 18.24 % and 1.20 %, respectively. MFP1P was mainly composed of →3)-α-D-Gal (1→, ß-D-Man-(1→ and →6)-α-D-Glc (1→ glycosidic bond and showed a spherical chain conformation with uniform distribution in solution. The MFP1P exhibited great antioxidant activity with oxygen-free radical absorption capacity (ORAC) values of 291.63±6.81 µmol TE/g and MDA IC50 of 0.289±0.022 mg/mL.

Glyco-Nanoadjuvants: Sugar Structures on Carriers of a Small Molecule TLR7 Ligand Affect Their Immunostimulatory Activities.

Toll-like receptors (TLRs) are pattern recognition receptors that activate innate immunity, and their ligands are promising adjuvants for vaccines and immunotherapies. Small molecule TLR7 ligands are ideal vaccine adjuvants as they induce not only proinflammatory cytokines but also type I interferons. However, their application has only been approved for local administration due to severe systemic immune-related adverse events. In a previous study, we prepared the gold nanoparticles coimmobilized with synthetic small molecule TLR7 ligand, 1V209, and α-mannose (1V209-αMan-GNPs). 1V209-αMan-GNPs were selectively delivered via a cell surface sugar-binding protein, mannose receptor, which enabled selective delivery of TLR7 ligands to immune cells. Besides the mannose receptor, immune cells express various sugar-binding proteins such as macrophage galactose binding lectins and sialic acid-binding immunoglobulin-type lectins and recognize distinct sugar structures. Hence, in the present study, we investigated whether sugar structures on GNPs affect the efficiency and selectivity of intracellular delivery and subsequent immunostimulatory potencies. Five neutral sugars and two sialosides were selected and each sugar was coimmobilized with 1V209 onto GNPs (1V209-SGNPs) and their innate immunostimulatory potencies were compared to that of 1V209-αMan-GNPs. The in vitro study using mouse bone marrow derived dendritic cells (BMDCs) demonstrated that α-glucose, α-N-acetylglucosamine, or α-fucose immobilized 1V209-SGNPs increased interleukin-6 and type I interferon release similar to that of 1V209-αMan-GNPs, whereas galacto-type sugar immobilized 1V209-SGNPs predominantly enhanced type I interferon release. In contrast, sialoside immobilized 1V209-SGNPs did not enhance the potency of 1V209. In the in vivo immunization study using ovalbumin as a model antigen, neutral sugar immobilized 1V209-SGNPs induced comparable T helper-1 immune response to that of 1V209-αMan-GNPs and by 10-fold higher than that of sialoside immobilized 1V209-SGNPs. These results indicate that the sugar structures on 1V209-SGNPs affect their immunostimulatory activities, and functionalization of the carrier particles is important to shape immune responses.

Mannose-Modified Liposome Co-Delivery of Human Papillomavirus Type 16 E7 Peptide and CpG Oligodeoxynucleotide Adjuvant Enhances Antitumor Activity Against Established Large TC-1 Grafted Tumors in Mice.

BACKGROUND: Previously, we demonstrated the therapeutic efficacy of a human papillomavirus (HPV) vaccine, including HPV16 E7 peptide and CpG oligodeoxynucleotides (CpG ODN), against small TC-1 grafted tumors. Here, we developed an HPV16 E7 peptide and CpG ODN vaccine delivered using liposomes modified with DC-targeting mannose, Lip E7/CpG, and determined its anti-tumor effects and influence on systemic immune responses and the tumor microenvironment (TME) in a mouse large TC-1 grafted tumor model. METHODS: L-alpha-phosphatidyl choline (SPC), cholesterol (CHOL), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycol-2000)] (DSPE-PEG-2000), 1,2-dioleoyl-3-trimethylammonium-propane chloride salt (DOTAP) and Mannose-PEG-DSPE, loaded with HPV16 E7 peptide and CpG ODN, were used to construct the Lip E7/CpG vaccine. The anti-tumor effects and potential mechanism of Lip E7/CpG were assessed by assays of tumor growth inhibition, immune cells, in vivo cytotoxic T lymphocyte (CTL) responses and cytokines, chemokines, CD31, Ki67 and p53 expression in the TME. In addition, toxicity of Lip E7/CpG to major organs was evaluated. RESULTS: Lip E7/CpG had a diameter of 122.21±8.37 nm and remained stable at 4°C for 7 days. Co-delivery of HPV16 E7 peptide and CpG ODN by liposomes exerted potent anti-tumor effects in large (tumor volume ≥200mm3) TC-1 grafted tumor-bearing mice with inhibition rates of 80% and 78% relative to the control and Free E7/CpG groups, respectively. Vaccination significantly increased numbers of CD4+ and CD8+ T cells, and IFN-γ-producing cells in spleens and tumors and enhanced HPV-specific CTL responses, while reducing numbers of inhibitory cells including myeloid-derived suppressor cells and macrophages. Expression of cytokines and chemokines was altered and formation of tumor blood vessels was reduced in the Lip E7/CpG group, indicating possible modulation of the immunosuppressive TME to promote anti-tumor responses. Lip E7/CpG did not cause morphological changes in major organs. CONCLUSION: Lip E7/CpG induced anti-tumor effects by enhancing cellular immunity and improving tumor-associated immunosuppression. Mannose-modified liposomes are the promising vaccine delivery strategy for cancer immunotherapy.

Optimization, characterization, rheological study and immune activities of polysaccharide from Sagittaria sagittifolia L.

To improve the extraction efficiency of polysaccharides from Sagittaria sagittifolia L. (SPU) by ultrasonic assisted extraction (UAE), the optimal extraction conditions were optimized as follows: extraction temperature of 85℃, extraction time of 15 min and ratio of liquid to raw material 43 mL/g, under these conditions, the yield of SPU increased by about 168 % compared with hot water extraction (HWE). After separation and purification by DEAE-52 cellulose column and Sephadex G-50 column, the pure polysaccharide fraction (SPU70-W1) was obtained, and its structure, rheology and immune activity were analyzed. The results indicated that SPU70-W1 (7.70 kDa) contained mannose, glucose and galactose in the molar ratio of 2.06:93.58:4.36 with typical pseudoplasticity fluids behavior and possessed the backbone of →2,6)-α-D-Glcp-(1→, →3,6)-α-D-Glcp-(1→, α-D-Glcp-(1→ and 6)-α-D-Glcp-(1→. In addition, SPU70-W1 exhibited remarkable immunomodulatory activity. Thus, SPU70-W1 could contribute to the food, medicine, cosmetics as a functional additive.

Immunomodulatory activity-guided isolation and characterization of a novel polysaccharide from Atractylodis macrocephalae Koidz.

The Rhizoma of Atractylodis macrocephala Koidz. is a traditional Chinese herbal medicine that has been widely and empirically used for unexplained recurrent spontaneous abortion. In this study, a polysaccharide from the Rhizoma of Atractylodis macrocephala Koidz., designated as RAMP2, with an absolute molecular weight of 4.354 × 103 Da was isolated and found to be composed of mannose, galacturonic acid, glucose, galactose and arabinose. The NMR results displayed that →3-ß-glcp-(1→, →3,6-ß-glcp-(1→, →6-ß-glcp-(1→, T-ß-glcp-(1→, →4-α-galpA-(1→, →4-α-galpA-6-OMe-(1→, →5-α-araf-(1→, →4,6-ß-manp-(1→ and →4-ß-galp-(1→ were the main linkages in RAMP2. TEM and SEM results indicated that RAMP2 was globular in structure. Furthermore, in vitro experiments on murine CD4+ T cells revealed that RAMP2 could increase the percentage of Treg cells, up-regulate Foxp3, IL-10 and IL-2 mRNA expressions and the secretion of IL-10 and IL-2. RAMP2 was further shown to increase STAT5 phosphorylation levels in Treg cells, suggesting that RAMP2 increased the number of Treg cells through IL-2/STAT5 pathway.

Multi-functional nanocomplex codelivery of Trp2 and R837 to activate melanoma-specific immunity.

Antigen-adjuvant combination could induce a protective and long-lasting anti-tumor immune response. However, exploiting system which could co-deliver melanoma antigen peptide Trp2 (Tyrosinase-related protein 2) and Toll-like-receptor-7 (TLR7) agonists imiquimod (R837) both are poor aqueous solubility is still challenging. Our new nanocomplex was explored for specific delivery of Trp2 and R837 into antigen-presenting cells (APCs). R837 was loaded into mannosylated-ß-cyclodextrin (Man-CD) to target dendritic cells (DCs) by binding mannose receptors (MR) on DCs. A fusion peptide (WT) was constructed by incorporating the amino acid region of TAT (cell-penetrating peptide) into Trp2 to improve the TAT-mediated intracellular efficiency. Negatively charged sodium alginate (SA), a biocompatible polymer, which can induce adjuvant responses by affecting the functions of DCs, was complexed with Man-CD/R837 and WT via physical adsorption. The optimized nanocomplex promoted the cellular uptake and showed remarkable efficacy to enhance the secreting of Th1-cytokines. This multi-functional nanocomplex system may allow effective targeting-codelivery of multi-hydrophobic drugs and be a promising subunit vaccine candidate as a potential prevention action of tumor.

Purification of GNA-Related Lectins from Natural Sources.

The Galanthus nivalis lectin, abbreviated as GNA, is the model protein for a large group of mannose-binding lectins. Here, we describe the purification of GNA starting from dry bulbs. Using a combination of ion exchange chromatography and affinity chromatography on mannose-Sepharose, a highly pure preparation of GNA can be obtained.

Construction and application of carbohydrate microarrays to detect foodborne bacteria.

The rapid detection of pathogenic bacteria is vital for the prevention of outbreaks of infectious diseases, including infections by the common foodborne bacteria E.coli and Salmonella Carbohydrate microarrays have been developed as a powerful method to investigate carbohydrate-protein interaction with only very small amounts of glycans, which show great potential for detect the carbohydrate mediated interaction with pathogens. Here, different mannose-coated microarrays were constructed and tested with E.coli (K-12 and BL-21) and Salmonella enterica strains (ATCC9184 and ATCC31685) exhibiting different mannose binding affinities. The optimized carbohydrate microarray was then applied to test the binding of 12 Salmonella enterica and 9 E.coli isolates from local patients for the first time and showed strong binding with certain serovars or subtypes. The results showed that microarray probed with the single mannose structure is not enough for the detection of bacteria with various serovars or subtypes, which contain a high degree of allelic variation in adhesin. We suggest that a complex carbohydrate microarray containing different glycan conformation may be needed for detection of different bacteria isolates.

Peptide Glycodendrimers as Potential Vaccines for Olive Pollen Allergy.

Olive pollen is one of the most important causes of respiratory allergy, with Ole e 1 being the most clinically relevant sensitizing allergen. Peptide-based vaccines represent promising therapeutic approaches, but the use of adjuvants is required to strengthen the weak immunogenicity of small peptides. We propose the use of dendrimeric scaffolds conjugated to the T cell immunodominant epitope of Ole e 1 (OE109-130) for the development of novel vaccines against olive pollen allergy. Four dendrimeric scaffolds containing an ester/ether with nine mannoses, an ester succinimidyl linker with nine N-acetyl-glucosamine units or nine ethylene glycol units conjugated to OE109-130 peptide were designed, and their cytotoxicity, internalization pattern, and immunomodulatory properties were analyzed in vitro. None of the dendrimers exhibited cytotoxicity in humanized rat basophil (RBL-2H3), human bronchial epithelial Calu-3, and human mast LAD2 cell lines. Confocal images indicated that mannosylated glycodendropeptides exhibited lower colocalization with a lysosomal marker. Moreover, mannosylated glycodendropeptides showed higher transport tendency through the epithelial barrier formed by Calu-3 cells cultured at the air-liquid interface. Finally, mannosylated glycodendropeptides promoted Treg and IL10+Treg proliferation and IL-10 secretion by peripheral blood mononuclear cells from allergic patients. Mannosylated dendrimers conjugated with OE109-130 peptide from Ole e 1 have been identified as suitable candidates for the development of novel vaccines of olive pollen allergy.

Structural characterization and antioxidant activities of a water soluble polysaccharide isolated from Glycyrrhiza glabra.

A water-soluble polysaccharide, named GPN, with molecular mass 38.7 kDa was isolated from Glycyrrhiza glabra with hot water extraction, ethanol precipitation, and purified by column chromatography. Monosaccharide composition analysis confirmed the presence of predominant glucose (98.03%) and trace amount of mannose, arabinose, and galactose. Methylation and GC-MS analysis revealed that the main glycosidic bonds in GPN comprised 1,4-linked Glcp, T-linked Glcp, 1,4,6-linked Glcp, and 1,6-linked Glcp. Based on these results and 1D/2D NMR spectroscopy, GPN has a linear backbone of 1,4-linked α-D-Glcp and 1,6-linked α-D-Glcp which substituted at C-4 of glucose. The side chain probably composed from 1,4-linked to main side α-D-Glcp and terminal 1-linked ß-D-Glcp to the C-3 of D residue. Congo red assay confirmed the existence of triple helix structure. Moreover, SEM and XRD analysis revealed that the GPN had irregular fibrous, filaments like surface; and both crystalline and amorphous structure. GPN also displayed favorable thermal stability. Moreover, G.glabra polysaccharide showed good antioxidant activity.

Isolations, characterizations and bioactivities of polysaccharides from the seeds of three species Glycyrrhiza.

In this paper, polysaccharides from the seeds of three species of genus Glycyrrhiza were extracted to investigate the physicochemical properties, structural characteristics and antioxidant activities. The polysaccharides were composed of xylose, mannose, galactose, and glucose with different molar ratio. Fourier transform infrared spectroscopy showed the presence of key functional groups of polysaccharides whereas scanning electron microscopy analysis revealed the characteristic morphology of different polysaccharides, and thermogravimetric analysis exhibited good thermal stability of all samples. The antioxidant activities of polysaccharides were evaluated in vitro. All the three polysaccharides demonstrated strong reducing power, as well as scavenging activity against DPPH, ABTS, and hydroxyl free radicals. Antioxidant assays indicated that all the polysaccharides have obvious antioxidant activities and possess a potential development and application value in food, cosmetics as well as pharmaceutical industries.

Gold Nanoparticles Coimmobilized with Small Molecule Toll-Like Receptor 7 Ligand and α-Mannose as Adjuvants.

Adjuvants enhance the immune response during vaccination. Among FDA-approved adjuvants, aluminum salts are most commonly used in vaccines. Although aluminum salts enhance humoral immunity, they show a limited effect for cell-mediated immune responses. Thus, further development of adjuvants that induce T-cell-mediated immune response is needed. Toll-like receptors (TLRs) recognizing specific pathogen-associated molecular patterns activate innate immunity, which is crucial to shape adaptive immunity. Using TLR ligands as novel adjuvants in vaccines has therefore attracted substantial attention. Among them a small molecule TLR7 ligand, imiquimod, has been approved for clinical use, but its use is restricted to local administration due to unwanted adverse side effects when used systematically. Since TLR7 is mainly located in the endosomal compartment of immune cells, efficient transport of the ligand into the cells is important for improving the potency of the TLR7 ligand. In this study we examined gold nanoparticles (GNPs) immobilized with α-mannose as carriers for a TLR7 ligand to target immune cells. The small molecule synthetic TLR7 ligand, 2-methoxyethoxy-8-oxo-9-(4-carboxy benzyl)adenine (1V209), and α-mannose were coimmobilized via linker molecules consisting of thioctic acid on the GNP surface (1V209-αMan-GNPs). The in vitro cytokine production activity of 1V209-αMan-GNPs was higher than that of the unconjugated 1V209 derivative in mouse bone marrow-derived dendritic cells and in human peripheral blood mononuclear cells. In the in vivo immunization study, 1V209-αMan-GNPs induced significantly higher titers of IgG2c antibody specific to ovalbumin as an antigen than did unconjugated 1V209, and splenomegaly and weight loss were not observed. These results indicate that 1V209-αMan-GNPs could be useful as safe and effective adjuvants for development of vaccines against infectious diseases and cancer.

Selective targeting of tumor cells and tumor associated macrophages separately by twin-like core-shell nanoparticles for enhanced tumor-localized chemoimmunotherapy.

Tumor associated macrophage (TAM)-based immunotherapy has been presented as a promising strategy in cancer therapy. The combination of TAM-based immunotherapy with sorafenib (SF) could be conceivably quite more effective in hepatocellular carcinoma (HCC) treatment. A co-delivery system was superior in improving the co-accumulation of two drugs in tumor tissues for chemoimmunotherapy, while in the case of selective targeting of separated cells such as tumor cells and immune cells, a novel targeted co-delivery strategy was badly required. In this study, twin-like core-shell nanoparticles (TCN) were developed for synchronous biodistribution and separated cell targeting delivery of SF and TAM re-polarization agents IMD-0354 to cancer cells and TAM to enhance tumor-localized chemoimmunotherapy, respectively. First of all, SF loaded cationic lipid-based nanoparticles (SF-CLN) and mannose-modified IMD-0354 loaded cationic lipid-based nanoparticles (M-IMD-CLN) were prepared, respectively. SF on the surface of SF-CLN and mannose on the M-IMD-CLN were regarded as targeting ligands for selective targeting delivery of SF-CLN and M-IMD-CLN to cancer cells and TAM separately. Then, pH-responsive charge reversal polymer O-carboxymethyl-chitosan (CMCS) was coated on the SF-CLN and M-IMD-CLN to obtain twin-like CMCS/SF-CLN and CMCS/M-IMD-CLN, respectively. The results of cellular uptake assay on Hepa1-6 cells and RAW 264.7 cells in vitro, respectively, as well as the results of tumor tissue distribution of SF and IMD-0354 in vivo suggested that CMCS/SF-CLN and CMCS/M-IMD-CLN exhibited similar properties in vitro and synchronous biodistribution in vivo, and were efficient at separated cell targeting delivery. What's more, the results of antitumor efficiency in vivo and phenotype analysis of TAM in tumor tissues proved that CMCS/SF-CLN and CMCS/M-IMD-CLN exhibited superior synergistic antitumor efficacy and M2-type TAM polarization ability compared with SF treatment in Hepa1-6 tumor bearing mice. Consequently, TCN which was the combination of co-administration and nano-drug delivery systems has great potential to be used in tumor-localized chemoimmunotherapy in clinics.

Proposed dual antagonist approach for the prevention and treatment of urinary tract infections caused by uropathogenic Escherichia coli.

Urinary tract infections are among the most common infectious diseases worldwide, primarily caused by uropathogenic Escherichia coli (UPEC) strains that harbor type I pili and P pili on the surface. Standard E. coli therapy still entails antibiotic consumption, but urinary tract infections tend to recur at a very high rate. Due to the emergence of antibiotic resistant strains of UPEC, as well as high infection recurrence rates, there is a need for new approaches to efficiently treat and prevent urinary tract infections. Since aforementioned adhesive organelles are the principal virulence factors in UPEC, anti-adhesion strategy seems to be the most promising (and hitherto unexplored) treatment option. Here we propose an antiadhesive dual targeting approach towards FimH and PapG adhesive proteins placed on two key virulence factors for UPEC - type I fimbriae and P pili. Such dual antagonists will contain appropriate pharmacophores (mannose and natural cranberry-containing polyphenol) joined together and will more efficiently block the infection and prevent the progression of the disease in comparison to FimH and PapG as isolated targets. More specifically, polyphenol mannosides (due to the structural similarities with the most potent biaryl inhibitors) can act as high-affinity FimH ligands, while cranberry-associated polyphenol moiety can additionally inhibit the PapG-mediated adhesion. Proposed compound may also contribute to the antioxidant capacity of the human organism. In conclusion, this dual-target hypothesis for the prevention and treatment of UPEC infections represents an important foundation for further research on this topic.