Emerging polymeric materials for treatment of oral diseases: design strategy towards a unique oral environment.

Oral diseases are prevalent but challenging diseases owing to the highly movable and wet, microbial and inflammatory environment. Polymeric materials are regarded as one of the most promising biomaterials due to their good compatibility, facile preparation, and flexible design to obtain multifunctionality. Therefore, a variety of strategies have been employed to develop materials with improved therapeutic efficacy by overcoming physicobiological barriers in oral diseases. In this review, we summarize the design strategies of polymeric biomaterials for the treatment of oral diseases. First, we present the unique oral environment including highly movable and wet, microbial and inflammatory environment, which hinders the effective treatment of oral diseases. Second, a series of strategies for designing polymeric materials towards such a unique oral environment are highlighted. For example, multifunctional polymeric materials are armed with wet-adhesive, antimicrobial, and anti-inflammatory functions through advanced chemistry and nanotechnology to effectively treat oral diseases. These are achieved by designing wet-adhesive polymers modified with hydroxy, amine, quinone, and aldehyde groups to provide strong wet-adhesion through hydrogen and covalent bonding, and electrostatic and hydrophobic interactions, by developing antimicrobial polymers including cationic polymers, antimicrobial peptides, and antibiotic-conjugated polymers, and by synthesizing anti-inflammatory polymers with phenolic hydroxy and cysteine groups that function as immunomodulators and electron donors to reactive oxygen species to reduce inflammation. Third, various delivery systems with strong wet-adhesion and enhanced mucosa and biofilm penetration capabilities, such as nanoparticles, hydrogels, patches, and microneedles, are constructed for delivery of antibiotics, immunomodulators, and antioxidants to achieve therapeutic efficacy. Finally, we provide insights into challenges and future development of polymeric materials for oral diseases with promise for clinical translation.

Development of Procathepsin L (pCTS-L)-Inhibiting Lanosterol-Carrying Liposome Nanoparticles to Treat Lethal Sepsis.

The pathogenesis of microbial infections and sepsis is partly attributable to dysregulated innate immune responses propagated by late-acting proinflammatory mediators such as procathepsin L (pCTS-L). It was previously not known whether any natural product could inhibit pCTS-L-mediated inflammation or could be strategically developed into a potential sepsis therapy. Here, we report that systemic screening of a NatProduct Collection of 800 natural products led to the identification of a lipophilic sterol, lanosterol (LAN), as a selective inhibitor of pCTS-L-induced production of cytokines [e.g., Tumor Necrosis Factor (TNF) and Interleukin-6 (IL-6)] and chemokines [e.g., Monocyte Chemoattractant Protein-1 (MCP-1) and Epithelial Neutrophil-Activating Peptide (ENA-78)] in innate immune cells. To improve its bioavailability, we generated LAN-carrying liposome nanoparticles and found that these LAN-containing liposomes (LAN-L) similarly inhibited pCTS-L-induced production of several chemokines [e.g., MCP-1, Regulated upon Activation, Normal T Cell Expressed and Presumably Secreted (RANTES) and Macrophage Inflammatory Protein-2 (MIP-2)] in human blood mononuclear cells (PBMCs). In vivo, these LAN-carrying liposomes effectively rescued mice from lethal sepsis even when the first dose was given at 24 h post the onset of this disease. This protection was associated with a significant attenuation of sepsis-induced tissue injury and systemic accumulation of serval surrogate biomarkers [e.g., IL-6, Keratinocyte-derived Chemokine (KC), and Soluble Tumor Necrosis Factor Receptor I (sTNFRI)]. These findings support an exciting possibility to develop liposome nanoparticles carrying anti-inflammatory sterols as potential therapies for human sepsis and other inflammatory diseases.

Systemic administration of choline acetyltransferase decreases blood pressure in murine hypertension.

Acetylcholine (ACh) decreases blood pressure by stimulating endothelium nitric oxide-dependent vasodilation in resistance arterioles. Normal plasma contains choline acetyltransferase (ChAT) and its biosynthetic product ACh at appreciable concentrations to potentially act upon the endothelium to affect blood pressure. Recently we discovered a T-cell subset expressing ChAT (TChAT), whereby genetic ablation of ChAT in these cells produces hypertension, indicating that production of ACh by TChAT regulates blood pressure. Accordingly, we reasoned that increasing systemic ChAT concentrations might induce vasodilation and reduce blood pressure. To evaluate this possibility, recombinant ChAT was administered intraperitoneally to mice having angiotensin II-induced hypertension. This intervention significantly and dose-dependently decreased mean arterial pressure. ChAT-mediated attenuation of blood pressure was reversed by administration of the nitric oxide synthesis blocker L-nitro arginine methyl ester, indicating ChAT administration decreases blood pressure by stimulating nitic oxide dependent vasodilation, consistent with an effect of ACh on the endothelium. To prolong the half life of circulating ChAT, the molecule was modified by covalently attaching repeating units of polyethylene glycol (PEG), resulting in enzymatically active PEG-ChAT. Administration of PEG-ChAT to hypertensive mice decreased mean arterial pressure with a longer response duration when compared to ChAT. Together these findings suggest further studies are warranted on the role of ChAT in hypertension.

An in situ tissue engineering scaffold with growth factors combining angiogenesis and osteoimmunomodulatory functions for advanced periodontal bone regeneration.

BACKGROUND: The regeneration of periodontal bone defect remains a vital clinical challenge. To date, numerous biomaterials have been applied in this field. However, the immune response and vascularity in defect areas may be key factors that are overlooked when assessing the bone regeneration outcomes of biomaterials. Among various regenerative therapies, the up-to-date strategy of in situ tissue engineering stands out, which combined scaffold with specific growth factors that could mimic endogenous regenerative processes. RESULTS: Herein, we fabricated a core/shell fibrous scaffold releasing basic fibroblast growth factor (bFGF) and bone morphogenetic protein-2 (BMP-2) in a sequential manner and investigated its immunomodulatory and angiogenic properties during periodontal bone defect restoration. The in situ tissue engineering scaffold (iTE-scaffold) effectively promoted the angiogenesis of periodontal ligament stem cells (PDLSCs) and induced macrophage polarization into pro-healing M2 phenotype to modulate inflammation. The immunomodulatory effect of macrophages could further promote osteogenic differentiation of PDLSCs in vitro. After being implanted into the periodontal bone defect model, the iTE-scaffold presented an anti-inflammatory response, provided adequate blood supply, and eventually facilitated satisfactory periodontal bone regeneration. CONCLUSIONS: Our results suggested that the iTE-scaffold exerted admirable effects on periodontal bone repair by modulating osteoimmune environment and angiogenic activity. This multifunctional scaffold holds considerable promise for periodontal regenerative medicine and offers guidance on designing functional biomaterials.

Enhanced Oral Bioavailability and Anti-Echinococcosis Efficacy of Albendazole Achieved by Optimizing the "Spring" and "Parachute".

Maximizing the pharmacological efficacy of albendazole (ABZ), an anti-echinococcosis drug, is essential in the long-term treatment of patients with echinococcosis. As a weakly alkaline drug, ABZ has a pH-dependent solubility that decreases dramatically from gastric fluid (pH 1.4) to intestinal fluid (pH 6.5), where it is absorbed. In this study, we endeavored to develop an optimized tablet formulation of ABZ to improve its dissolution and oral bioavailability from two aspects: a faster initial dissolution in the gastric pH condition (i.e., the "spring") and a more prolonged drug supersaturation in the intestinal pH condition (i.e., the "parachute"). To achieve this goal, ABZ-HCl salt was selected first, which demonstrated a higher intrinsic dissolution rate under pH 1.4 compared with the ABZ free base that is used in the commercial product Albenda. Second, by comparing the ABZ supersaturation kinetics under pH 6.5 in the presence of various polymers including poly(vinylpyrrolidone) (PVP), PVP/VA, hydroxypropyl methylcellulose (HPMC), and HPMC acetate succinate (HPMC-AS), HPMC-AS was found to be the most effective crystallization inhibitor for ABZ, likely due to the hydrophobic interaction between ABZ and HPMC-AS in an aqueous environment. The newly designed tablet formulation containing ABZ-HCl and HPMC-AS showed ∼3 times higher oral bioavailability compared with that of Albenda in Beagle dogs. More significantly, the anti-echinococcosis efficacy of the improved formulation was 2.4 times higher than that of Albenda in a secondary hepatic alveolar echinococcosis Sprague-Dawley rat model. The strategy of simultaneously improving the spring and parachute of an oral formulation of ABZ, by using a highly soluble salt and an effective polymeric crystallization inhibitor, was once again proven to be a viable and readily translatable approach to optimize the unsatisfactory oral medicines due to solubility and bioavailability limitations.

Enhanced osteogenesis of quasi-three-dimensional hierarchical topography.

Natural extracellular matrices (ECMs) are three-dimensional (3D) and multi-scale hierarchical structure. However, coatings used as ECM-mimicking structures for osteogenesis are typically two-dimensional or single-scaled. Here, we design a distinct quasi-three-dimensional hierarchical topography integrated of density-controlled titania nanodots and nanorods. We find cellular pseudopods preferred to anchor deeply across the distinct 3D topography, dependently of the relative density of nanorods, which promote the osteogenic differentiation of osteoblast but not the viability of fibroblast. The in vivo experimental results further indicate that the new bone formation, the relative bone-implant contact as well as the push-put strength, are significantly enhanced on the 3D hierarchical topography. We also show that the exposures of HFN7.1 and mAb1937 critical functional motifs of fibronectin for cellular anchorage are up-regulated on the 3D hierarchical topography, which might synergistically promote the osteogenesis. Our findings suggest the multi-dimensions and multi-scales as vital characteristic of cell-ECM interactions and as an important design parameter for bone implant coatings.

Promoting the Outgrowth of Neurites on Electrospun Microfibers by Functionalization with Electrosprayed Microparticles of Fatty Acids.

Controlling the outgrowth of neurites is important for enhancing the repair of injured nerves and understanding the development of nervous systems. Herein we report a simple strategy for enhancing the outgrowth of neurites through a unique integration of topographical guidance and a chemical cue. We use electrospray to easily functionalize the surface of a substrate with microparticles of natural fatty acids at a controllable density. Through a synergistic effect from the surface roughness arising from the microparticles and the chemical cue offered by the fatty acids, the outgrowth of neurites from PC12 cells is greatly enhanced. We also functionalize the surfaces of uniaxially aligned, electrospun microfibers with the microparticles and further demonstrate that the substrates can guide and enhance directional outgrowth of neurites from both PC12 multicellular spheroids and chick embryonic dorsal root ganglia bodies.

Antibiotic cement-coated locking plate as a temporary internal fixator for femoral osteomyelitis defects.

PURPOSE: Recently we modified the Masquelet technique by using an antibiotic cement-coated locking plate as a temporary internal fixator when treating septic bone defects. This modification is in order to prevent the complications related to external fixator use and provides the involved limb with a greater stability to undergo earlier and more vigorous physical therapy for recovery of joint function. The purpose of this study was to assess the outcomes of large femoral osteomyelitis defects managed by Masquelet technique combined with the antibiotic cement-coated locking plate used as a temporary internal fixator. METHODS: Between November 2013 to November 2014, 13 cases of large femoral osteomyelitis defects were treated by Masquelet technique and the antibiotic cement-coated locking plate was used as a temporary internal fixator in the first stage surgery. All the patients' clinical and imaging results were retrospectively analyzed. RESULTS: After debridement, there was a femoral bone defect with a mean of 9.8 cm (range, 5-16 cm). The mean follow-up was 17.8 months (range 12 to 24 months). One patient developed infection in nine months after second stage surgery. Radiographic bony union was achieved within a mean 20.3 weeks (range, 18-30 weeks) in all patients. The mean time period to full weight bearing after the second step procedure was 5.8 months (range, 5-8.5 months). The mean knee range of motion for the patients at the last follow up was 122° (range 100-135°). CONCLUSION: Based on our experience, we believe that antibiotic cement-coated locking plate is a viable fixation method in the first stage of Masquelet technique for the management of large femoral osteomyelitis defects. It may offer a better chance of infection eradication as well as improved recovery of joint function without increasing the infection recurrence rate and without compromising bone graft union.

Micropatterning of the Ferroelectric Phase in a Poly(vinylidene difluoride) Film by Plasmonic Heating with Gold Nanocages.

Polymer thin films with patterned ferroelectric domains are attractive for a broad range of applications, including the fabrication of tactile sensors, infrared detectors, and non-volatile memories. Herein, we report the use of gold nanocages (AuNCs) as plasmonic nanostructures to induce a ferroelectric-paraelectric phase transition in a poly(vinylidene fluoride) (PVDF) thin film by leveraging its photothermal effect. This technique allows us to generate patterned domains of ferroelectric PVDF within just a few seconds. The incorporation of AuNCs significantly enhances the pyroelectric response of the ferroelectric film under near-infrared irradiation. We also demonstrate the use of such patterned ferroelectric films for near-infrared sensing/imaging.

Nanostructured titanate with different metal ions on the surface of metallic titanium: a facile approach for regulation of rBMSCs fate on titanium implants.

Titanium (Ti) is widely used for load-bearing bio-implants, however, it is bio-inert and exhibits poor osteo-inductive properties. Calcium and magnesium ions are considered to be involved in bone metabolism and play a physiological role in the angiogenesis, growth, and mineralization of bone tissue. In this study, a facile synthesis approach to the in situ construction of a nanostructure enriched with Ca(2+) and Mg(2+) on the surface of titanium foil is proposed by inserting Ca(2+) and Mg(2+) into the interlayers of sodium titanate nanostructures through an ion-substitution process. The characteriz 0.67, and 0.73 nm ation results validate that cations can be inserted into the interlayer regions of the layered nanostructure without any obvious change of morphology. The cation content is positively correlated to the concentration of the solutions employed. The biological assessments indicate that the type and the amount of cations in the titanate nanostructure can alter the bioactivity of titanium implants. Compared with a Na(+) filled titanate nanostructure, the incorporation of divalent ions (Mg(2+) , Ca(2+) ) can effectively enhance protein adsorption, and thus also enhance the adhesion and differentiation ability of rat bone-marrow stem cells (rBMSCs). The Mg(2+) /Ca(2+) -titanate nanostructure is a promising implantable material that will be widely applicable in artificial bones, joints, and dental implants.

[Comparisons of therapeutic efficacy and safety of solifenacin versus tolterodine in patients with overactive bladder: a meta-analysis of outcomes].

OBJECTIVE: To compare the clinical efficacy and safety of solifenacin versus tolterodine in patients with overactive bladder (OAB). METHODS: Literature searches were performed with PubMed, EMBASE, Ovid and Google Scholar databases, Wanfang and CNKI from inception to October 2013 for comparative studies assessing solifenacin and tolterodine for OAB. The data were extracted and evaluated by two reviewers independently according to the Cochrane Handbook for Systematic Reviews. Meta-analyses were conducted with RevMan 5.2. RESULTS: A total of 7 studies involving 1 805 patients were retrieved. Compared with tolterodine immediate release (IR), the number of urgency episodes and urge incontinence episodes in 24 h and the rate of dry mouth were significantly lower in patients on solifenacin (RR = -0.34, 95% CI: -0.50--0.18, P = 0.00; RR = -0.29, 95% CI:-0.55--0.04, P = 0.03; RR = 0.58, 95% CI: 0.41-0.83, P = 0.00) and the rate of constipation was higher in those on solifenacin (RR = 2.72, 95% CI: 1.38-5.39, P = 0.00). No significant differences existed between tolterodine IR and solifenacin in mean micturition volume per voiding and micturitions episodes in 24 h (P = 0.05,0.08). Between solifenacin and tolterodine extended release (ER), the number of urgency episodes, micturition and urge incontinence episodes in 24 h and mean micturition volume per voiding were not statistically different (all P > 0.05). The incidence of major adverse events, such as dry mouth, constipation and blurred vision, was not significantly different (all P > 0.05). And most adverse events were mild. CONCLUSIONS: Solifenacin is superior to tolterodine IR in treating OAB symptoms. However the rate of constipation is higher for solifenacin. Both solifenacin and tolterodine ER have similar therapeutic efficacies and adverse events.

[Clinical effectiveness and safety of combined therapy with alpha-blocker and an anticholinergic drug for bladder outlet obstruction with overactive bladder: a Meta-analysis of outcomes].

OBJECTIVE: To compare the clinical effectiveness and safety of alpha-blocker alone and combined tamsulosin with an anticholinergic drug for bladder outlet obstruction (BOO) with overactive bladder (OAB). METHODS: Literature search was performed using PubMed, EMBASE, Ovid, Wanfang, and CNKI from inception to October 2013 for comparative studies assessing alpha-blocker alone and combined alpha-blocker with an anticholinergic drug for BOO+OAB. Data were extracted and evaluated by two reviewers independently according to the Cochrane Handbook for systematic reviews. Meta-analyses were conducted using RevMan 5.2. RESULTS: A total of 7 studies involving 3 458 patients were included for the analysis. The values of total IPSS and storage IPSS reduced significantly after treatment in combination group (RR = -0.23, 95%CI: -0.44--0.02, P = 0.03; RR = -0.69, 95%CI: -0.88--0.51, P < 0.01). There were no significant differences between the two groups in voiding IPSS and Qmax (P = 0.86 and 0.89). The incidences of dry mouth (OR = 2.53), constipation (OR = 3.74), dizziness (OR = 0.73), and urinary retention (OR = 0.26) were higher in combination group than in alpha-blocker alone group (P < 0.05). But most adverse events were mild in degree. CONCLUSION: Alpha-blocker combined with an anticholinergic drug in the treatment of BOO+OAB was better than that of alpha-blocker alone, and was safe and well tolerated.

Microenvironment-Driven Fenton Nanoreactor Enabled by Metal-Phenolic Encapsulation of Calcium Peroxide for Effective Control of Dental Caries.

Caries are one of the most common oral diseases caused by pathogenic bacterial infections, which are widespread and persistently harmful to human health. Using nanoparticles to invade biofilms and produce reactive oxygen species (ROS) in situ is a promising strategy for killing bacteria and disrupting the structure of biofilms. In this work, a biofilm-targeting Fenton nanoreactor is reported that can generate ROS responsive to the cariogenic microenvironment. The nanoreactor is constructed by metal-phenolic encapsulation of calcium peroxide (CaO2) followed by modification with a biofilm targeting ligand dextran. Within the cariogenic biofilm, the Fenton nanoreactor is activated by an acidic microenvironment to be decomposed into H2O2 and iron ions, triggering a Fenton-like reaction to generate ROS that can eliminate the biofilm by breaking down extracellular polymeric substances (EPS) and killing cariogenic bacteria. Meanwhile, the depletion of excess protons in biofilm leads to a reversal of the cariogenic microenvironment. The Fenton nanoreactor can effectively inhibit the biofilm formation of Streptococcus mutans on ex vivo human teeth and is effective in preventing caries meanwhile maintaining the oral microbial diversity in rat caries infection model. This work provides a novel and efficient modality for acid microenvironment-driven ROS therapy.

Using Mendelian randomization provides genetic insights into potential targets for sepsis treatment.

Sepsis is recognized as a major contributor to the global disease burden, but there is a lack of specific and effective therapeutic agents. Utilizing Mendelian randomization (MR) methods alongside evidence of causal genetics presents a chance to discover novel targets for therapeutic intervention. MR approach was employed to investigate potential drug targets for sepsis. Pooled statistics from IEU-B-4980 comprising 11,643 cases and 474,841 controls were initially utilized, and the findings were subsequently replicated in the IEU-B-69 (10,154 cases and 454,764 controls). Causal associations were then validated through colocalization. Furthermore, a range of sensitivity analyses, including MR-Egger intercept tests and Cochran's Q tests, were conducted to evaluate the outcomes of the MR analyses. Three drug targets (PSMA4, IFNAR2, and LY9) exhibited noteworthy MR outcomes in two separate datasets. Notably, PSMA4 demonstrated not only an elevated susceptibility to sepsis (OR 1.32, 95% CI 1.20-1.45, p = 1.66E-08) but also exhibited a robust colocalization with sepsis (PPH4 = 0.74). According to the present MR analysis, PSMA4 emerges as a highly encouraging pharmaceutical target for addressing sepsis. Suppression of PSMA4 could potentially decrease the likelihood of sepsis.

AI model to detect contact relationship between maxillary sinus and posterior teeth.

Objectives: To establish a novel deep learning networks (MSF-MPTnet) based on panoramic radiographs (PRs) for automatic assessment of relationship between maxillary sinus floor (MSF) and maxillary posterior teeth (MPT), and to compare accuracy of MSF-MPTnet, dentists and radiologists identifying contact relationship. Study design: A total of 1035 PRs and 1035 Cone-beam computed tomographys （CBCT）images were collected from January 2018 to April 2022. The relationships were classified into class I and II by CBCT. Class I represents non-contact group, and class II represents contact group. 350 PRs were randomly selected as test dataset and accuracy of MSF-MPTnet, dentists, and radiologists was compared. Results: The intraclass correlation coefficient of dentists was 0.460-0.690 and it was 0.453-0.664 for radiologists. Sensitivity and accuracy of MSF-MPTnet were 0.682-0.852and 0.890-0.951, indicating that the output performance of MSF-MPTnet was reliable. Accuracy of maxillary premolars and molars were 79.7%-90.3 %, 76.2%-89.2 % and 72.9%-88.3 % in MSF-MPTnet model, dentists and radiologists. Accuracy of class I relationship in the MSF-MPTnet model (67.7%-94.6 %) was higher than that of dentists (56.5%-84.6 %) in maxillary first premolars and right second premolar, and accuracy of class I relationship in the MSF-MPTnet model is also higher than radiologists (40.0%-78.1 %) in all teeth positions (p < 0.05). Conclusions: MSF-MPTnet model could increase detecting accuracy of the relationship between MSF and MPT, minimize pseudo contact relationship and reduce frequency of CBCT use.

A Multifunctional Metal-Phenolic Nanocoating on Bone Implants for Enhanced Osseointegration via Early Immunomodulation.

Surface modification is an important approach to improve osseointegration of the endosseous implants, however it is still desirable to develop a facile yet efficient coating strategy. Herein, a metal-phenolic network (MPN) is proposed as a multifunctional nanocoating on titanium (Ti) implants for enhanced osseointegration through early immunomodulation. With tannic acid (TA) and Sr2+ self-assembled on Ti substrates, the MPN coatings provided a bioactive interface, which can facilitate the initial adhesion and recruitment of bone marrow mesenchymal stem cells (BMSCs) and polarize macrophage toward M2 phenotype. Furthermore, the TA-Sr coatings accelerated the osteogenic differentiation of BMSCs. In vivo evaluations further confirmed the enhanced osseointegration of TA-Sr modified implants via generating a favorable osteoimmune microenvironment. In general, these results suggest that TA-Sr MPN nanocoating is a promising strategy for achieving better and faster osseointegration of bone implants, which can be easily utilized in future clinical applications.

Discovering medical conditions associated with periodontitis using linked electronic health records.

AIM: To use linked electronic medical and dental records to discover associations between periodontitis and medical conditions independent of a priori hypotheses. MATERIALS AND METHODS: This case-control study included 2475 patients who underwent dental treatment at the College of Dental Medicine at Columbia University and medical treatment at NewYork-Presbyterian Hospital. Our cases are patients who received periodontal treatment and our controls are patients who received dental maintenance but no periodontal treatment. Chi-square analysis was performed for medical treatment codes and logistic regression was used to adjust for confounders. RESULTS: Our method replicated several important periodontitis associations in a largely Hispanic population, including diabetes mellitus type I (OR = 1.6, 95% CI 1.30-1.99, p < 0.001) and type II (OR = 1.4, 95% CI 1.22-1.67, p < 0.001), hypertension (OR = 1.2, 95% CI 1.10-1.37, p < 0.001), hypercholesterolaemia (OR = 1.2, 95% CI 1.07-1.38, p = 0.004), hyperlipidaemia (OR = 1.2, 95% CI 1.06-1.43, p = 0.008) and conditions pertaining to pregnancy and childbirth (OR = 2.9, 95% CI: 1.32-7.21, p = 0.014). We also found a previously unreported association with benign prostatic hyperplasia (OR = 1.5, 95% CI 1.05-2.10, p = 0.026) after adjusting for age, gender, ethnicity, hypertension, diabetes, obesity, lipid and circulatory system conditions, alcohol and tobacco abuse. CONCLUSIONS: This study contributes a high-throughput method for associating periodontitis with systemic diseases using linked electronic records.

Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides.

In view of the biological significance and micro-heterogeneity of protein glycosylation for human health, specific enrichment of N-glycosylated proteins/peptides from complex biological samples is a prerequisite for the discovery of disease biomarkers and clinical diagnosis. In this work, we propose a "grafting-from" N-glycoprotein enriching method based on the in-situ growth of thermoresponsive polymer brushes from the N-glycosylated site of proteins. The initiator was first attached to the pre-oxidized glycan moieties by hydrazide chemistry, from which the thermoresponsive polymers can be grown to form giant protein-polymer conjugates (PPC). The thermosensitive PPC can be precipitated and separated by raising the temperature to above its lower critical solubility temperature (LCST). Mass spectrometry verified 210 N-glycopeptides corresponding to 136 N-glycoproteins in the rabbit serum. These results demonstrate the capability of the tandem thermoprecipitation strategy to enrich and separate N-glycoprotein/glycopeptide. Due to its simplicity and efficiency specifically, this method holds the potential for identifying biomarkers from biological samples in N-glycoproteome analysis.

A potential role for Giardia chaperone protein GdDnaJ in regulating Giardia proliferation and Giardiavirus replication.

BACKGROUND: Giardia duodenalis (referred to as Giardia) is a flagellated binucleate protozoan parasite, which causes one of the most common diarrheal diseases, giardiasis, worldwide. Giardia can be infected by Giardiavirus (GLV), a small endosymbiotic dsRNA virus belongs to the Totiviridae family. However, the regulation of GLV and a positive correlation between GLV and Giardia virulence is yet to be elucidated. METHODS: To identify potential regulators of GLV, we performed a yeast two-hybrid (Y2H) screen to search for interacting proteins of RdRp. GST pull-down, co-immunoprecipitation and bimolecular fluorescence complementation (BiFC) assay were used to verify the direct physical interaction between GLV RdRp and its new binding partner. In addition, their in vivo interaction and colocalization in Giardia trophozoites were examined by using Duolink proximal ligation assay (Duolink PLA). RESULTS: From Y2H screen, the Giardia chaperone protein, Giardia DnaJ (GdDnaJ), was identified as a new binding partner for GLV RdRp. The direct interaction between GdDnaJ and GLV RdRp was verified via GST pull-down, co-immunoprecipitation and BiFC. In addition, colocalization and in vivo interaction between GdDnaJ and RdRp in Giardia trophozoites were confirmed by Duolink PLA. Further analysis revealed that KNK437, the inhibitor of GdDnaJ, can significantly reduce the replication of GLVs and the proliferation of Giardia. CONCLUSION: Taken together, our results suggested a potential role of GdDnaJ in regulating Giardia proliferation and GLV replication through interaction with GLV RdRp.

Surface-Confined Piezocatalysis Inspired by ROS Generation of Mitochondria Respiratory Chain for Ultrasound-Driven Noninvasive Elimination of Implant Infection.

Implant-associated infections (IAI) are great challenges to medical healthcare and human wellness, yet current clinical treatments are limited to the use of antibiotics and physical removal of infected tissue or the implant. Inspired by the protein/membrane complex structure and its generation of reactive oxygen species in the mitochondria respiration process of immune cells during bacteria invasion, we herein propose a metal/piezoelectric nanostructure embedded on the polymer implant surface to achieve efficient piezocatalysis for combating IAI. The piezoelectricity-enabled local electron discharge and the induced oxidative stress generated at the implant-bacteria interface can efficiently inhibit the activity of the attachedStaphylococcus aureusby cell membrane disruption and sugar energy exhaustion, possess high biocompatibility, and eliminate the subcutaneous infection by simply applying the ultrasound stimulation. For further demonstration, the treatment of root canal reinfection with simplified procedures has been achieved by using piezoelectric gutta-percha implanted in ex vivo human teeth. This surface-confined piezocatalysis antibacterial strategy, which takes advantage of the limited infection interspace, easiness of polymer processing, and noninvasiveness of sonodynamic therapy, has potential applications in IAI treatment.