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The aquaculture industry is vital in providing a valuable protein food source for humans, but generates a huge amount of solid and dissolved wastes that pose great risks to the environment and aquaculture sustainability. Suspended solids (in short SS), one of the aquaculture wastes, are very difficult to be treated due to their high organic contents. The bioconversion from wastewater, food effluents, and activated sludge into poly-β-hydroxybutyrate (PHB) is a sustainable alternative to generate an additional income and could be highly attractive to the agricultural and environmental management firms. However, little is known about its potential application in aquaculture wastes. In the present study, we first determined that 7.2% of SS was PHB. Then, the production of PHB was increased two-fold by the optimal fermentation conditions of wheat bran and microbial cocktails at a C/N ratio of 12. Also, the PHB-enriched SS showed a higher total ammonia nitrogen removal rate. Importantly, we further demonstrated that the PHB-enriched SS as a feed could promote fish growth and up-regulate the expression of the immune-related genes. Our study developed an eco-friendly and simple approach to transforming problematic SS wastes into PHB-enriched high-quality food for omnivorous fish, which will increase the usage efficiency of SS and provide a cheaper diet for aquatic animals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Qiao, Li, Li, Zhang, Shen, Zhao, Zhang, Qi, Chen, Sun, Cang, Liu, Wangkahart and Wang.)

Bcl2-associated athanogene3 (BAG3) protein, mainly induced by stressful stimuli, has been confirmed to participate in apoptosis and autophagy. In recent studies, BAG3 has gradually become a key molecule in tumors. However, the role of BAG3 in the progression of lumbar facet joint osteoarthritis (FJOA) and whether it can regulate chondrocyte apoptosis and autophagy are still unknown. In both human and FJOA rat models, we observed an upregulation of BAG3 and apoptosis and autophagy-related proteins compared with healthy tissues. Then, we established the chondrocytes injury model in vitro by using IL-1β to stimulate human SW1353 cells. Western blot analysis data showed significant expression of BAG3, apoptosis, and autophagy-related proteins in SW1353 cells. Finally, by knocking down and overexpressing BAG3, we discovered possible anti-apoptotic and autophagy-promoted effects of BAG3 in FJOA through various experimental methods. This study demonstrated that BAG3 actively participates in regulating chondrocyte apoptosis and autophagy in FJOA and may be a highly interesting target for pharmacological interventions., (© 2022. The Author(s) under exclusive licence to University of Navarra.)

Large-scale and diffuse population flow amplifies the localized COVID-19 outbreak into a widespread pandemic. Network analysis provides a new methodology to uncover the topology and evolution of the population flow and understand its influence on the early dynamics of COVID-19 transmission. In this paper, we simulated 42 transmission scenarios to show the distribution of the COVID-19 outbreak across China. We predicted some original (Guangzhou, Shanghai, Shenzhen) had higher total aggregate population outflows than Wuhan, indicating larger spread scopes and faster growth rates of COVID-19 outbreak. We built an importation risk model to identify some major cities (Dongguan and Foshan) with the highest total importation risk values and the highest standard deviations, indicating the core transmission chains (Dongguan-Shenzhen, Foshan-Guangzhou). We built the population flow networks to analyze their Spatio-temporal characteristics and identify the influential sub-groups and spreaders. By removing different influential spreaders, we identified Guangzhou can most influence the network's topological characteristics, and some major cities' degree centrality was significantly decreased. Our findings quantified the effectiveness of travel restrictions on delaying the epidemic growth and limiting the spread scope of COVID-19 in China, which helped better derive the geographical COVID-19 transmission related to population flow networks' structural features., Competing Interests: None., (© 2021 Elsevier Ltd. All rights reserved.)

Tumor necrosis factor receptor-associated factor 6 (TRAF6), a regulator of NF-κB signaling, has been discovered recently to be probably related to osteoarthritis, while the function of TRAF6 in lumbar facet joint osteoarthritis(FJOA)still remains unknown. The aim of this study was to probe the specific function of TRAF6 in chondrocytes and its connection with the pathophysiology of FJOA. We found upregulation of TRAF6 in FJOA cartilage by western blot analysis. In vitro, we stimulated immortalized human chondrocytes by LPS to establish the cells apoptosis model. Western blot analysis demonstrated that levels of TRAF6 and cleaved caspase-3/8 in the chondrocyte injury model increased significantly. Knockdown of TRAF6 suppressed the expression of matrix metallopeptidase-13 (MMP-13) and interleukin-6 (IL-6) induced by LPS, and alleviated cell apoptosis. Meanwhile, western blot and immunofluorescent staining demonstrated that IκBα degradation and p65 nuclear transportation were also inhibited, revealing that knockdown of TRAF6 suppressed activation of the NF-κB pathway in LPS-induced chondrocytes apoptosis model. Collectively, our findings suggest that TRAF6 plays a crucial role in FJOA development by regulating NF-κB signaling pathway. Knockdown of TRAF6 may supply a potential therapeutic strategy for FJOA.

This study reports a simple and practical method to introduce antimicrobial and biofilm-controlling functions into hydroxyl- or amino-containing polymers such as cellulose using compounds derived from widely used reactive dyes. Two dichloro- s -triazine-based dyes, reactive blue 4 and sodium 4-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate (a colorless reactive "dye"), were covalently attached to cellulose at room temperature by replacing one chloride on the dyes with the hydroxyl groups on cellulose followed by hydrolysis under alkaline conditions to transform the remaining chloride into hydroxyl groups. The chemical reactions were confirmed by FT-IR studies, energy-dispersive X-ray spectroscopy, water contact angle measurement, and zeta potential analysis. The resulting cellulose provided powerful antimicrobial activities against Staphylococcus epidermidis ( S. epidermidis , ATCC 35984, Gram-positive bacteria), Escherichia coli ( E. coli , ATCC 15597, Gram-negative bacteria), and Candida albicans ( C. albicans , ATCC 10231, yeast) and effectively prevented the formation of bacterial or fungal biofilms. The minimum inhibition concentrations of the hydrolyzed dyes were similar to that of phenol. In the zone of inhibition studies using phenolic compounds as positive controls, the hydrolyzed dyes and their model compound cyanuric acid demonstrated antimicrobial functions, suggesting that the antimicrobial activities were associated with the phenol-like hydroxyl groups on the triazine rings. Antimicrobial mechanism investigation indicated that the phenol-like structures on the dyed cellulose caused microbial lysis and leakage of intracellular components. The antimicrobial functions were durable upon repeated washing, and the dyed cellulose showed outstanding biocompatibility toward mammalian cells.

Background: Spinal cord injuries (SCIs), along with subsequent secondary injuries, often result in irreversible damage to both sensory and motor functions. However, a thorough view of the underlying pathological mechanisms of SCIs, especially in a temporal-spatial manner, is still lacking., Methods: To obtain a comprehensive, real-time view of multiple subsets of the cellular mechanisms involved in SCIs, we applied RNA-sequencing technology to characterize the temporal changes in gene expression around the lesion site of contusion SCI in rats. First, we identified the differentially expressed genes (DEGs) in contrast to sham controls at 1, 4, and 7 days post SCI. Through bioinformatics analysis, including Pathway analysis, Gene-act-net, and Pathway-act-net, we screened and verified potential key pathways and genes associated with either the acute or subacute stages of SCI pathology., Results: The top three overrepresented pathways were associated with cytokine-cytokine receptor interaction, TNF signaling pathway, and cell cycle at day 1; lysosome, cytokine-cytokine receptor interaction, phagosome at day 4; and phagosome, lysosome, cytokine-cytokine receptor interaction at day 7 post injury. Further, we identified uniquely enriched genes at each time point, such as Ccr1 and Nos2 at day 1; as well as Mgst2, and Pla2g3 at 4 and 7 days post-injury., Conclusions: Our pathway analysis suggested a transition from inflammatory responses to multiple forms of cell death processes from the acute to subacute stages of SCI. Further, our results revealed a continuous transformation from a more inflammatory to an apoptotic/self-repairing transcriptome following the time-course of SCIs. Our research provides novel insights into the molecular mechanisms of SCI pathophysiology and identifies potential targets for therapeutic intervention after SCI., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/atm-20-6519). The authors have no conflicts of interest to declare., (2020 Annals of Translational Medicine. All rights reserved.)

The pathophysiological changes in cartilage are a crucial feature of lumbar facet joint (LFJ) degeneration and arthritis. However, the molecular mechanism of human LFJ degeneration remains largely defined. This study aimed to examine the changes in chondrocytes at different stages of degenerative LFJ using hematoxylin and eosin and Safranin O staining. The significant loss of chondrocytes in grades 2 and 3 of LFJs was observed. The expression levels of CCAAT enhancer binding protein β (C/EBPβ), Runt-related transcription factor 2 (RUNX2), and matrix metalloproteinase 13 (MMP13) also increased with the aggravation of degeneration (4.89, 5.77, and 6.3 times by Western blot). In vitro, chondrocytes scraped from the LFJs during surgery were stimulated by interleukin (IL)-1β to establish the injury model. The association of C/EBPβ and RUNX2 with active caspase-3 on chondrocytes was analyzed. The high expression level of C/EBPβ, RUNX2, and MMP13 was consistent with that of caspase-3, which reached a peak after 36 h of stimulation. Immunofluorescence suggested that C/EBPβ, RUNX2, and MMP13 co-labeled with active caspase-3. Moreover, immunoprecipitation data prompted that C/EBPβ was able to interact with RUNX2. The knockdown of C/EBPβ significantly decreased the expression levels of MMP13 and active caspase-3 (2.48 and 2.89 times as detected by Western blot analysis) and inhibited chondrocyte apoptosis, which was further demonstrated using flow cytometry. Taken together, the findings of this study uncovered that C/EBPβ could interact with RUNX2 to induce chondrocyte apoptosis in human LFJ degeneration by regulating the expression of MMP13.

Facet joint osteoarthritis (FJOA) is a common degenerative joint disorder with high prevalence in the elderly. FJOA causes lower back pain and lower extremity pain, and thus severely impacts the quality of life of affected patients. Emerging studies have focused on the histomorphological and histomorphometric changes in FJOA. However, the dynamic genetic changes in FJOA have remained to be clearly determined. In the present study, previously obtained RNA deep sequencing data were subjected to an ingenuity pathway analysis (IPA) and canonical signaling pathways of differentially expressed genes (DEGs) in FJOA were studied. The top 25 enriched canonical signaling pathways were identified and canonical signaling pathways with high absolute values of z-scores, specifically leukocyte extravasation signaling, Tec kinase signaling and osteoarthritis pathway, were investigated in detail. DEGs were further categorized by disease, biological function and toxicity (tox) function. The genetic networks between DEGs as well as hub genes in these functional networks were also investigated. It was demonstrated that C-X-C motif chemokine ligand 8, elastase, neutrophil expressed, growth factor independent 1 transcriptional repressor, Spi-1 proto-oncogene, CCAAT enhancer binding protein epsilon, GATA binding protein 1, TAL bHLH transcription factor 1, erythroid differentiation factor, minichromosome maintenance complex component 4, BTG anti-proliferation factor 2, BRCA1 DNA repair-associated, cyclin D1, chromatin assembly factor 1 subunit A, triggering receptor expressed on myeloid cells 1 and tumor protein p63 were hub genes in the top 5 IPA networks (with a score >30). The present study provides insight into the pathological processes of FJOA from a genetic perspective and may thus benefit the clinical treatment of FJOA., (Copyright: © Chen et al.)

Osteoarthritis is a general joint disease characterized by articular cartilage degeneration. The extracellular matrix is a principal component in articular cartilage. The dynamic remodeling of the extracellular matrix is involved in the pathological degradation of the articular cartilage. Facet joint osteoarthritis (FJOA) is a common form of osteoarthritis that occurs in the posterior aspect of the vertebral column. However, to the best of our knowledge, the current understanding of the genetic changes in FJOA is limited. The most significantly differentially expressed genes and Gene Ontology categories in FJOA were identified by transcriptome sequencing analysis. The extracellular matrix, matrix metalloproteinases (MMPs) and proteinases of the extracellular matrix were highly involved in FJOA. The canonical signaling pathway 'inhibition of matrix metalloproteinases' was further studied in detail by identifying and validating differentially expressed genes in the signaling pathway. Taken together, the present study revealed changes in MMP-related genes in FJOA and showed the importance of extracellular matrix remodeling in FJOA from a genetic aspect., (Copyright: © Chen et al.)

Background: Osteoarthritis (OA) is a prevalent degenerative joint disease, which was characterized by inflammation and cartilage degradation. Accumulating evidence has demonstrated that Tanshinone I has an anti-inflammatory effect in various diseases. However, the efficacy of Tanshinone I as an anti-inflammatory agent in OA remains unclear. This study aimed to explore the role of Tanshinone I on OA both in vitro and in vivo., Methods: CHON-001 cells were treated with IL-1β (10 ng/mL) for 72 hrs to induce OA model in vitro. Meanwhile, CHON-001 cells were pre-treated with 20 μM Tanshinone I for 24 hrs and then stimulated with IL-1β (10 ng/mL) for 72 hrs. CCK-8, immunofluorescence and flow cytometry assays were used to detect the viability, proliferation and apoptosis in CHON-001 cells, respectively. Western blotting assay was used to detect the levels of collagen II, aggrecan, MMP-13, cleaved caspase 1, Gasdermin D, SOX11 and p-NF-κB in CHON-001 cells. In addition, the mouse model of OA was built by anterior cruciate ligament transection (ACLT) in the right knee. Meanwhile, the mice were administrated with 10 or 30 mg/kg Tanshinone I for 8 weeks. Safranin-O/Fast Green staining was used to assess cartilage destruction in a mouse model of OA., Results: In this study, IL-1β significantly induced apoptosis, extracellular matrix degradation and inflammatory response in CHON-001 cells. Tanshinone I significantly inhibited IL-1β-induced apoptosis in CHON-001 cells. In addition, the IL-1β-induced collagen II, aggrecan degradation, SOX11 downregulation, and MMP-13 and p-NF-κB upregulation in CHON-001 cells were notably reversed by Tanshinone I treatment. Moreover, Tanshinone I alleviated cartilage destruction and synovitis and reduced OARSI scores and subchondral bone thickness in a mouse model of OA., Conclusion: Our findings showed that Tanshinone I could alleviate the progression of OA in vitro and in vivo. These results demonstrated that Tanshinone I might be regarded as a promising therapeutic agent for the treatment of OA., Competing Interests: The authors report no conflicts of interest in this work., (© 2019 Wang et al.)

Dexamethasone (DEX) induces significant cytotoxicity to human osteoblasts. cPWWP2A is recently-indentified novel circular RNA (circRNA), acting as an endogenous sponge of microRNA-579 (miR-579). The present study tested the expression and potential functions of the cPWWP2A-miR-579 axis in DEX-treated osteoblasts. We show that cPWWP2A is downregulated in the necrotic femoral head tissues of DEX-taking human patients as well as in DEX-treated human osteoblasts. In OB-6 osteoblastic cells and primary human osteoblasts ectopic overexpression of cPWWP2A potently inhibited DEX-induced miR-579 accumulation, cell death, apoptosis and programmed necrosis. Silencing miR-579, by targeted siRNAs, also attenuated DEX-induced cytotoxicity in human osteoblasts. Significantly, mimicking DEX-induced actions, cPWWP2A silencing or forced miR-579 overexpression induced significant cytotoxicity in human osteoblasts. Further analyses demonstrated that miR-579's targets, including SIRT1 and PDK1 (phosphoinositide-dependent protein kinase 1), were downregulated in DEX-treated osteoblasts. Their levels were decreased as well in the necrotic femoral head tissues of DEX-taking human patients. Taken together we show that dysregulation of the cPWWP2A-miR-579 axis is involved in DEX-induced cytotoxicity in human osteoblasts., (Copyright © 2019 Elsevier Inc. All rights reserved.)

In order to prepare multifunctional fibrous membranes with hydrophobicity, antibacterial properties and UV resistance, we used silica and titanium dioxide for preparing SiO 2 @TiO 2 nanoparticles (SiO 2 @TiO 2 NPs) to create roughness on the fibrous membranes surfaces. The introduction of TiO 2 was used for improving UV resistance. N-Halamine precursor and silane precursor were introduced to modify SiO 2 @TiO 2 NPs to synthesize SiO 2 @TiO 2 -based core@shell composite nanoparticles. The hydrophobic antibacterial fibrous membranes were prepared by a dip-pad process of electrospun biodegradable polyhydroxybutyrate/poly-ε-caprolactone (PHB/PCL) with the synthesized SiO 2 @TiO 2 -based core@shell composite nanoparticles. TEM, SEM and FT-IR were used to characterize the synthesized SiO 2 @TiO 2 -based core@shell composite nanoparticles and the hydrophobic antibacterial fibrous membranes. The fibrous membranes not only showed excellent hydrophobicity with an average water contact angle of 144° ± 1°, but also appreciable air permeability. The chlorinated fibrous membranes could inactivate all S. aureus and E. coli O157:H7 after 5 min and 60 min of contact, respectively. In addition, the chlorinated fibrous membranes exhibited outstanding cell compatibility with 102.1% of cell viability. Therefore, the prepared hydrophobic antibacterial degradable fibrous membranes may have great potential application for packaging materials., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

The reactions of a monomeric N-halamine, 1-chloro-5,5-dimethylhydantoin (MCDMH), and a mesoporous material-supported N-halamine (MMSNs) with phenol and p-cresol (two common contaminants in water) were investigated. MCDMH reacted rapidly with the phenolic compounds, and pH values had little effects on the reactions. On the contrary, MMSNs reacted with phenol and p-cresol only when the pH values were higher than 10. Phenol showed a lower reaction rate than p-cresol toward MMSNs. GCMS analysis suggested that MMSNs might react with the phenolic compounds through step-wise electrophilic chlorination reactions, and the main product was 2,4,6-trichlorophenol. The reaction kinetics were studied by following the disappearance of phenolic UV absorption bands, and the kinetic parameters were determined., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Formation of biofilms on solid surfaces is a long-standing challenge to multiple medical and health-related applications. Once formed, biofilms are very difficult to destroy, and microorganisms in biofilms are much more resistant to antimicrobial agents than their free-floating counterparts. The current antimicrobial agents/disinfectants often have low residence time on biofilms, leading to low antimicrobial effect on biofilm microorganisms. We designed and synthesized an amphiphilic quaternary ammonium chitosan (CS612) as biocompatible antimicrobial agents that bind onto preformed biofilms to kill adherent microorganisms. CS612 was synthesized through an external acid-free approach, and showed excellent concentration-dependent cytocompatibility toward mammalian cells. Antimicrobial functions of CS612 were confirmed by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests against Gram-positive bacteria, Gram-negative bacteria, and fungi. CS612 rapidly bound onto preformed bacterial and fungal biofilms, and killed adherent microorganisms living in the biofilms. The biofilm-binding kinetic parameters were determined, pointing to a new strategy to manage microorganisms in biofilms and their accompanying problems., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Antimicrobial mesoporous materials with polymer brushes on the surface were prepared, and their structure and antimicrobial performance investigated. Poly ((3-acrylamidopropyl) trimethylammonium chloride) (PAPTMAC) modified mesoporous silica was prepared by a polymer-brush-grafted method through treatment with the initiator 4,4'-azobis (4-cyanovaleric acid) (ACVA) and polymerized with (3-acrylamidopropyl) trimethylammonium chloride (APTMAC). A covalent bond was formed between mesoporous silica and N-halamine precursor; N-H bonds were successfully transformed to N-Cl bonds after chlorination. Morphology and structure of mesoporous silica were affected to some extent after modification. The surface area of the polymerized sample decreased, but was sufficient for further applications. Compare to the original sample, antimicrobial properties of the polymerized samples with quaternary ammonium groups (QAS) increased slightly. After exposure to dilute household bleach, the chlorinated samples showed excellent antimicrobial properties against 100% of S. aureus (ATCC 6538) (7.63 log) and E. coli O157:H7 (ATCC 43895) (7.52 log) within 10 min. The prepared mesoporous silicas with effective antimicrobial properties could be very useful for potential application in water filtration., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Oral ulcerative lesions are painful and debilitating, particularly for immunosuppressed patients undergoing chemotherapeutic/irradiation treatment. Their clinical management requires multiple drugs to be administered simultaneously. Current formulations available to patients require frequent dosing, leading to poor compliance and suboptimal clinical outcomes. In this study, we prepared wheat germ agglutinin (WGA)-conjugated liposomes (WGA liposomes) to serve as bioadhesive drug carriers that can encapsulate various classes of drugs, rapidly bind to oral epithelial cells within minutes, and stay on the cells to provide sustained, localized drug release for days. Fluorescence binding studies found a significant increase ( p < 0.05) in the binding of WGA liposomes to oral cells in as short an incubation time as 1 min compared to that for nonconjugated liposomes. WGA liposomes encapsulating model drug amoxicillin showed sustained in vitro drug release, and the released drugs provided potent antimicrobial activity against Streptococcus mutans in an oral epithelial-bacterial coculture system. Exocytosis studies confirmed that the WGA liposomes stayed within the oral cells for 48 h, after which the cells completely removed the liposomes. Moreover, cell viability studies showed that there was a significant reduction in oral cell damage when the bacterially infected cells were treated with amoxicillin-loaded WGA liposomes compared to the untreated controls. These results point to the great potential of the lectin-conjugated liposomes as cell-binding drug-delivery systems in achieving localized, sustained drug release for the management of oral ulcerative lesions and other related complications.

HS1-associated protein X-1 (HAX1) is a class of multifunctional protein, participated in various physiological processes such as cell apoptosis, proliferation and motility. However, the HAX1 expression and function in the spinal cord injury (SCI) pathological process have not been investigated. In our current research, the rat model of SCI was established, and then we explored the possible role of HAX1 after SCI. The results of western blot indicated that HAX1 was present in sham operated control group and significantly elevated at 3 days post SCI, then declined gradually. Immunohistochemical studies indicated HAX1 expression was enhanced significantly in white and gray matter at 3 days post SCI compared with sham operated group. Double immunofluorescence staining showed the proportion of cells, double-labeled HAX1 and neurons, astrocytes, increased significantly at 3 days post SCI. In addition, co-localization of HAX1/active caspase-3 and HAX1/PCNA was tested in cells. Furthermore, over-expression of HAX1 inhibited neuronal apoptosis in vitro, and in astrocytes HAX1 silencing could down-regulate PCNA expression post LPS treatment. Meanwhile, CCK8 assay showed that knockdown of HAX1 could inhibit the astrocyte proliferation. In summary, our data indicated that HAX1 might play significant roles in pathological process of neuronal apoptosis and astrocyte proliferation during SCI., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Tumor necrosis factor receptor-associated factor 2 (TRAF2) has been demonstrated that it plays a significant role in cell death receptor signal transduction. The purpose of this study was to investigate the expression of TRAF2 and its possible role in FJOA. We observed an up-regulation of TRAF2 in FJOA by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) compared to normal tissues. In vitro, we used TNF-α to stimulate Human SW1353 chondrosarcoma cells to establish the chondrocytes injury model. Western blot analysis revealed significant expression of TRAF2 and cleaved caspase-3/8 in SW1353 cells. Co-localization of TRAF2/cleaved caspase-3/8 was detected in the cells injury model by double-labeling immunofluorescent staining. We demonstrated a possible anti-apoptotic effect of TRAF2 in chondrocyte apoptosis in FJOA by knockdown of its expression with siRNA. Moreover, TRAF2 knockdown was demonstrated to enhance TNF-α-induced apoptosis by flow cytometry assay. In conclusion, our results show that the up-regulation of TRAF2 may play an important role in the inhibition of chondrocyte apoptosis of FJOA., (Copyright © 2018 Elsevier Inc. All rights reserved.)

This article describes an electrochemical method to remove bacterial biofilm from a stainless steel (SS) surface using a potential pulse/reverse pulse technique. This technique employs a periodic waveform that consists of anodic and cathodic pulses. The pulses can effectively strip a thin layer of metal off the SS surface, along with the adherent biofilm, in a saline solution. Not only can the pulses effectively remove biofilm from the SS surface, but they also regenerate the original mirror-like shiny surface. The importance of this electrochemical biofilm removal method is its wide applicability for any types of biofilms. That is, instead of directly removing the biofilm, it removes a very thin layer of the metal under the biofilm. Thus, the removal process is independent to the nature of the biofilms. Furthermore, this electrochemical biofilm removal method is rapid (less than 30 s of potential pulse time) and does not require hazardous chemicals.

E3 ubiquitin ligase c-Caritas B cell lymphoma (c-cbl) is associated with negative regulation of receptor tyrosine kinases, signal transduction of antigens and cytokine receptors, and immune response. However, the expression and function of c-cbl in the regulation of neuropathic pain after chronic constriction injury (CCI) are unknown. In rat CCI model, c-cbl inhibited the activation of spinal cord microglia and the release of pro-inflammatory factors including tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β) and interleukin 6 (IL-6), which alleviated mechanical and heat pain through down-regulating extracellular signal-regulated kinase (ERK) pathway. Additionally, exogenous TNF-α inhibited c-cbl protein level vice versa. In the primary microglia transfected with c-cbl siRNA, when treated with TNF-α or TNF-α inhibitor, the corresponding secretion of IL-1β and IL-6 did not change. In summary, CCI down-regulated c-cbl expression and induced the activation of microglia, then activated microglia released inflammatory factors via ERK signaling to cause pain. Our data might supply a novel molecular target for the therapy of CCI-induced neuropathic pain.

A series of amine-based cationic polymeric N-halamine precursors, poly(2,2,6,6-tetramethyl-4-piperidyl methacrylate-co-trimethyl-2-methacryloxyethylammonium chloride)(PMPQ), were synthesized by copolymerizing 2,2,6,6-tetramethyl-4-piperidyl methacrylate (TMPM) with trimethyl-2-methacryloxyethylammonium chloride (TMAC) at different molar ratios (TMPM:TMAC = 10:90,30:70,50:50,70:30, and 90:10). After chlorine bleach treatment, the TMPM moieties in the new copolymers were transformed into amine-based N-halamines (Cl-PMPQ). The chemical structures of the samples were characterized with 1 H NMR, FT-IR, and UV spectra, and the molecular weights were determined by dynamic light scattering (DLS). With lower than 70 mol% of the original TMPM content, the resulting Cl-PMPQ copolymers were soluble in water, and demonstrated potent antibacterial functions against Escherichia coli (E. coli, a representative Gram-negative bacteria) and Staphylococcus epidermidis (S. epidermidis, a representative Gram-positive bacteria). E. coli and S. epidermidis were allowed to form biofilms on glass slides. Zeta potential analyses demonstrated that the Cl-PMPQ copolymers rapidly adsorbed onto the preexisting biofilms, and bacterial culturing studies confirmed that the bound Cl-PMPQ provided a total kill of the adherent bacteria in the biofilms. The kinetics of the Cl-PMPQ binding onto the preexisting biofilms were studied with UV analyses. The data fitted well to the bimodal model. The binding kinetic parameters of Cl-PMPQ onto the bacterial biofilms were thus determined., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Facet joint osteoarthritis is common lumbar osteoarthritis characterized by facet joint cartilage degeneration. However, the molecular basis of facet joint osteoarthritis remains largely undetermined. In the current study, we collected facet joint tissue samples from 10 control patients and 48 patients with facet joint osteoarthritis (20 patients with moderate degeneration and 28 with severe degeneration). The control patients underwent internal fixation of the lumbar spine due to vertebral fracture. RNA deep sequencing was performed, and Bioinformatic tools were applied. Among top 30 enriched signaling pathways, we focused on two inflammation-related signaling pathways, Wnt and NF-κB signaling pathways. Subsequently, using the quantitative RT-PCR analysis, we confirmed that in Wnt signaling pathway, the mRNA levels of Dickkopf WNT Signaling Pathway Inhibitor 2 (DKK2), Sex-determining Region Y-box 17 (SOX17), MYC, Cyclin D1, Calcium/Calmodulin Dependent Protein Kinase II Alpha (CAMK2A), and Wnt Family Member 11 and 5 were increased in facet joint osteoarthritis, while the mRNA levels of WNT Inhibitory Factor 1, Casein Kinase 1 Alpha 1, Transcription Factor 7/Lymphoid Enhancer Binding Factor 1 (TCF7/LEF1), and VANGL Planar Cell Polarity Protein 2 were decreased. In NF-κB signaling pathway, the mRNA levels of C-C Motif Chemokine Ligand 4 (CCL4) and C-C Motif Chemokine Ligand 4 Like 2 (CCL4L2) were increased, while the mRNA levels of BCL2 Related Protein A1 were decreased. These results suggest that Wnt and NF-κB signaling may be altered in the process of facet joint cartilage degeneration. The present study will expand our understanding of the molecular bases underlying facet joint osteoarthritis.

Although thought as a selective serotonin reuptake inhibitor (SSRI), the antidepressant mechanisms of paroxetine remain unknown. Previous studies have shown the role of the mammalian target of rapamycin (mTOR) signaling in depression. In this study, we investigated whether the antidepressant effects of paroxetine require mTOR signaling. We first examined whether chronic paroxetine administration restores the effects of CUMS and CSDS on the mTOR signaling cascade in the hippocampus and prefrontal cortex. Then, the pharmacologcial inhibitors of mTOR signaling (LY294002, U0126 and rapamycin) were used to assay if the paroxetine-induced reversing effects in the CUMS and CSDS models were prevented by mTOR system blockade. Furthermore, gene knockdown of mTOR by mTOR-shRNA was also used to test whether mTOR is necessary for the antidepressant effects of paroxetine. It was found that paroxetine treatment fully reversed the effects of CUMS and CSDS on the mTOR signaling in the hippocampus, but not the prefrontal cortex. Pharmacological inhibition of the mTOR signaling significantly blocked the antidepressant effects of paroxetine in the CUMS and CSDS models. Moreover, gene silencing of hippocampal mTOR by mTOR-shRNA also abolished the antidepressant effects of paroxetine. Taken together, hippocampal mTOR signaling is necessary for the antidepressant effects of paroxetine., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Vimentin, among the family of the intermediate filament, plays as the organizer of some critical proteins involved in migration, attachment, and cell signaling. In this study, the role of vimentin in chronic constriction injury (CCI) was investigated. Western blot revealed increased protein level of vimentin following CCI, peaking at 7 days. Double immunofluorescent staining showed that vimentin was mostly co-localized with astrocytes, not with neurons or microglia. In vitro, sensory neuronal injury stimulated astrocytes to produce more pro-inflammation cytokines, p-ERK (phosphorylated extracellular signal-regulated protein kinase), and vimentin. However, vimentin knockdown by siRNA (small interfering RNA) reversed the upregulation of p-ERK and vimentin expression and reduced the release of pro-inflammatory cytokines. Overall, stimulated astrocytes might release pro-inflammatory cytokines to promote the development of neuropathic pain via vimentin/ERK signaling.

Bacterial colonization and biofilm formation on dental resin composites cause secondary caries that shortens the service life of dental restorative materials. In this study, commercial barium borosilicate based glass powders, fillers widely used in dental composites, were covalently immobilized with silver sulfadiazine to provide antimicrobial effects. The chemical reactions were followed by FT-IR study, elemental analysis, and energy dispersive spectroscopy (EDS), and the reaction pathway was confirmed by model compound studies. The resulting glass powders demonstrated potent antimicrobial effects against Streptococcus mutans (S. mutans). BisGMA (2,2-bis[4-(2-hydroxy-3-methacrylyloxypropoxy)phenyl] propane)-based dental resins containing 2% to 10% of the new silver sulfadiazine glass powders exhibited powerful and durable antimicrobial efficacy against S. mutans. The mechanical properties of the antimicrobial resins were not negatively affected by the silver sulfadiazine glass powders, making them attractive candidates as antimicrobial fillers for dental composites and other related biomedical applications., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Annexin A2 (ANXA2), is a member of the annexin family of proteins that exhibit Ca 2+ -dependent binding to phospholipids. One attractive biological function of ANXA2 is participating in DNA synthesis and cell proliferation. Previous studies have shown that ANXA2 play a role in the development of the central nervous system. However, the biological function of ANXA2 after spinal cord injury (SCI) is still with limited acquaintance. In the present study, we performed a SCI model in adult rats and investigated the dynamic changes of ANXA2 expression in the spinal cord. Western blot analysis indicated a striking expression upregulation of ANXA2 after SCI. Immunohistochemistry further confirmed that ANXA2 immunoactivity was expressed at low levels in normal condition and increased at 5day after SCI. Double immunofluorescence staining prompted that ANXA2 immunoreactivity was found in astrocytes and neurons. Interestingly, ANXA2 expression was increased predominantly in astrocytes. We also examined the expression profiles of proliferating cell nuclear antigen (PCNA), Cyclin D1 and active caspase-3 in the injured spinal cords by western blot. Co-expression of ANXA2/PCNA, ANXA2/Cyclin D1 was detected in glial fibrillary acidic protein. Importantly, double immunofluorescence staining revealed that cell proliferation evaluated by PCNA appeared in many ANXA2-expressing cells and rare caspase-3 was observed in ANXA2-expressing cells after SCI. In addition, ANXA2 knockdown in astrocytes resulted in the increase of PCNA expression after LPS stimulation, showing that ANXA2 inhibited astrocyte proliferation after inflammation. Our data suggested that ANXA2 might play important roles in CNS pathophysiology after SCI., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study aims to compare two kinds of modified poly(lactic acid)(PLA)materials:PLA-chitosan(PLA-CTS)and PLA-poly(glycolic acid)(PLA-PGA).PLA-CTS and PLA-PGA scaffolds were prepared and observed under electron microscope.The scaffold porosity was calculated and the pH of the degradation solution was measured.Then rat olfactory ensheathing cells(OECs)were cultivated,and mixed cultured respectively with two scaffolds as two groups.The proliferation,adhesion rate and growth condition of the OECs were observed and compared between the two groups.Results showed that both the prepared PLA-CTS and PLA-PGA scaffolds were threedimensional porous structure and the porosity of PLA-CTS was 91%,while that of PLA-PGA was 87%.The pH of degradation solution decreased gradually,of which PLA-PGA fell faster than PLA-CTS.After added to the two scaffolds,most OECs could grow well,and there were no significant differences between the two groups on MTT test and nuclei number determined by fluorescent microscope.However,the cell adhesion rate of PLA-CTS group was significantly higher than that of PLA-PGA.It can be concluded that compared with PLA-PGA,PLA-CTS might be a better choice as OECs scaffold.

Fungal biofilm formation on healthcare materials is a significant clinical concern, often leading to medical-device-related infections, which are difficult to treat. A novel fungal repellent strategy is developed to control fungal biofilm formation. Methylacrylic acid (MAA) is grated onto poly methyl methacrylate (PMMA)-based biomaterials via plasma-initiated grafting polymerization. A cationic polymer, trimethylchitosan (TMC), is synthesized by reacting chitosan with methyl iodide. Sodium alginate (SA) is used as an anionic polymer. TMC/SA multilayers are coated onto the MAA-grafted PMMA via layer-by-layer self-assembly. The TMC/SA multilayer coatings significantly reduce fungal initial adhesion, and effectively prevent fungal biofilm formation. It is concluded that the anti-adhesive property of the surface is due to its hydrophilicity, and that the biofilm-inhibiting action is attributed to the antifungal activity of TMC as well as the chelating function of TMC and SA, which may have acted as fungal repellents. Phosphate buffered saline (PBS)-immersion tests show that the biofilm-modulating effect of the multilayer coatings is stable for more than 4 weeks. Furthermore, the presence of TMC/SA multilayer coatings improves the biocompatibility of the original PMMA, offering a simple, yet effective, strategy for controlling fungal biofilm formation., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Previous studies have found inconsistent results from testing methods used to measure heterotrophic plate count (HPC) bacteria in dental unit waterline (DUWL) samples. This study used 63 samples to compare the results obtained from an in-office chairside method and 2 currently used commercial laboratory HPC methods (Standard Methods 9215C and 9215E). The results suggest that the Standard Method 9215E is not suitable for application to DUWL quality monitoring, due to the detection of limited numbers of heterotrophic organisms at the required 35°C incubation temperature. The results also confirm that while the in-office chairside method is useful for DUWL quality monitoring, the Standard Method 9215C provided the most accurate results.

We reported a 12-year-old boy with neck pain and dysphagia 6 days after his neck being tied by another boy's arms from the back. The boy was found to have a tear-drop fracture of the axis in both radiograph and MRI, which was a rare case in children. He was placed in a cervical traction with Glisson belt and received transfusion for 2 weeks until his neck pain and dysphagia were released. Thereafter, an operation of anterior fusion and fixation was chosen to ensure further stability of the cervical spine and replacement of the fracture fragment. An 8-year follow-up examination showed a satisfactory range of motion of the cervical spine, fusion at C2-3 intervertebral space, and normal signal of C3-4 intervertebral disk. We conclude that tear-drop fractures of the axis in children may be caused by extension injury without a high-energy trauma, and operation of anterior fusion and fixation can be the treatment of choice.

Hypoxia and other adverse conditions are usually encountered by rapidly growing cells. The RNA-binding motif protein 3 (RBM3) is induced by low temperature and hypoxia. However, its expression and function in spinal cord injury are still unclear. To investigate the certain expression and biological function in the central nervous system, we performed an acute spinal cord contusion injury (SCI) model in adult rats. Western blot analysis indicated a striking expression upregulation of RBM3 after spinal cord injury (SCI). Double immunofluorescence staining prompted that RBM3 immunoreactivity was found in astrocytes and neurons. Interestingly, RBM3 expression was increased predominantly in astrocytes. Furthermore, colocalization of RBM3 with proliferating cell nuclear antigen (PCNA) was detected in astrocytes. To further understand whether RBM3 plays a role in astrocyte proliferation, we applied lipopolysaccharide (LPS) to induce astrocyte proliferation in vitro. Western blot analysis demonstrated that RBM3 expression was positively correlated with PCNA expression following LPS stimulation. Immunofluorescence analysis showed that the expression of RBM3 was also changed following the stimulation of astrocytes with LPS, which was parallel with the data in vivo. Additionally, knocking RBM3 down with small interfering RNA (siRNA) demonstrated that RBM3 might play a significant role in the proliferation of astrocytes treated by hypoxia in vitro. These results suggest that RBM3 may be involved in the proliferation of astrocytes after SCI. To summarize, we firstly uncover the temporal and spatial expression changes of RBM3 in spinal cord injury. Our data suggest that RBM3 might be implicated in central nervous system pathophysiology after SCI.

The performance of 2 American Public Health Association standard laboratory methods, the R2A spread plate and the SimPlate(TM) for heterotrophic plate count, for quantifying heterotrophic microorganisms in dental waterline samples was evaluated. Microbial counts were underestimated on SimPlate(TM) compared with R2A, and the results indicated a poor correlation between the 2 methods., (© 2013.)

Background: More and more attention has been focused on the inflammation or degeneration caused by biochemical factors in radiculopathy during lumbar facet joint degeneration. This study was designed to examine the expression and relationship of MMP-1/TIMP-1 and interleukin-1β (IL-1β), and to analyze the possible mechanism in degenerative lumbar facet joint disease., Methods: Lumbar facet joint cartilage and synovial tissues in 36 cases of posterior lumbar surgery were harvested to investigate IL-1β and MMP-1/TIMP-1 by immunohistochemistry and Western blot analysis. Double labeling immunofluorescence and real-time PCR, respectively, were used to assess the relationship between IL-1β and MMP-1., Results: IL-1β and MMP-1 were low in the lumbar disc herniation (LDH) group, and increased markedly in the lumbar spinal canal stenosis (LSCS) group (P < 0.05). However, there is no significant difference of TIMP-1 between LDH group and LSCS group (P > 0.05). Double staining results indicated that IL-1β overlapped with MMP-1 in the LSCS group. Moreover, real-time PCR results showed that MMP-1 mRNA in chondrocytes in vitro was affected in a dose- and time-dependent manner in response to IL-1β stimulation., Conclusions: Overexpression of MMP-1, induced by IL-1β, plays an important role in the inflammatory process of lumbar facet joint degeneration.

Minichromosome maintenance protein 7 (MCM7), a member of the minichromosome maintenance protein family, is essential for eukaryotic DNA replication initiation and the early stage of the elongation process. MCM7 participates in the cell cycle control of genome duplication. While it is ubiquitously expressed in all tissues, the biological function of MCM7 in the central nervous system is still with limited acquaintance. In the present study, we performed a spinal cord injury (SCI) model in adult rats. Western blotting indicated a marked alteration of MCM7 after SCI. Immunohistochemistry analysis revealed a wide distribution of MCM7 in the spinal cord. Double immunofluorescence staining showed that MCM7 immunoreactivity was increased predominantly in neurons, astrocytes, and microglia after SCI. We also examined the expression profiles of active caspase-3, proliferating cell nuclear antigen (PCNA), and Ki67, whose changes were correlated with the expression profiles of MCM7. Moreover, colocalization of MCM7/active caspase-3 was detected in neuronal nuclei (NeuN), and colocalization of MCM7/PCNA was detected in NeuN, glial fibrillary acidic protein, and CD11b, respectively. Our results suggest that MCM7 might be implicated in the apoptosis of neuron and proliferation of astrocytes and microglia after SCI.

In this study, a berberine-loaded electrospun poly-(epsilon-caprolactone) nanofibrous membrane (B-NFM) was formulated. The B-NFM was composed of uniform nanofibers with the average diameter of 190 +/- 53 nm. Whole blood assay revealed that the B-NFM had higher absorption capacity than gauze wound dressing (GWD) and non-drug-loaded nanofibrous membrane (NFM). Hemostatic studies showed that the B-NFM was more efficient at promoting blood clotting than GWD and NFM. B-NFM was also demonstrated to possess broad-spectrum antimicrobial activity. Biocompatibility studies indicated that B-NFM had good compatibility with skin fibroblast cells. These results suggested that the B-NFM can promote in vitro hemostasis to minimize wound bleeding, absorb biofluid to provide a favorable environment, inhibit on-site microbial agents to prevent wound infection, and support skin cell growth to facilitate wound healing. These data highly suggest a potential clinical use of B-NFM as wound dressing material.

Myelin and lymphocyte protein (MAL) was identified as a tetraspan proteolipid that is highly expressed by oligodendrocytes and Schwann cells as a component of compact myelin. It has also been reported to be a tumour suppressor and induces apoptosis through the Fas pathway. However, its expression and function in spinal cord injury are still unclear, especially in gray matter. In this study, we performed a spinal cord contusion injury (SCI) model in adult rats and detected the dynamic changes of MAL expression in spinal cord. Western blot and immunohistochemistry analysis revealed that MAL was present in gray and white matter of normal spinal cord. It gradually increased, got a peak at 1 day, and then declined to basal levels after spinal cord injury. Double immunofluorescence staining showed that MAL immunoreactivity was found in neurons and oligodendrocytes. Interestingly, MAL expression was increased predominantly in neurons rather than oligodendrocytes. We also examined the expression profiles of active caspase-3, whose changes were correlated with the expression profiles of MAL. Moreover, co-localization of MAL with active caspase-3 was detected. In conclusion, this is the first description of MAL expression changes in gray matter after spinal cord injury. Our results prompted that MAL might participate in CNS pathophysiology after SCI.

An N-halamine precursor, 5,5-dimethylhydantoin (DMH), was covalently linked to the surface of polyurethane (PU) with 1,6-hexamethylene diisocyanate (HDI) as the coupling agent. The reaction pathways were investigated using propyl isocyanate (PI) as a model compound. The results suggested that the imide and amide groups of DMH have very similar reactivities toward the isocyanate groups on PU surfaces activated with HDI. After bleach treatment the covalently bound DMH moieties were transformed into N-halamines. The new N-halamine-based PU provided potent antimicrobial effects against Staphylococcus aureus (Gram-positive bacterium), Escherichia coli (Gram-negative bacterium), methicillin-resistant Staphylococcus aureus (MRSA, drug-resistant Gram-positive bacterium), vancomycin-resistant Enterococcus faecium (VRE, drug-resistant Gram-positive bacterium), and Candida albicans (fungus), and successfully prevented bacterial and fungal biofilm formation. The antimicrobial and biofilm controlling effects were stable for longer than 6 months under normal storage in open air. Furthermore, if the functions were lost due to prolonged use they could be recharged by another chlorination treatment. The recharging could be repeated as needed to achieve long-term protection against microbial contamination and biofilm formation., (Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Glucose-regulated protein (GRP) 94 is a member of the stress protein family, which is localized in the endoplasmic reticulum (ER). Spinal cord injury (SCI) induced ER stress that results in apoptosis. However, the role of GRP94 in injury of the central nervous system remains unknown. In this study, we performed SCI in adult rats and investigated acutely the protein expression and cellular localization of GRP94 in the spinal cord. Western blot analysis revealed that GRP94 was low in normal spinal cord. It rose at 6h after SCI, peaked at 1 day, remained for another 3 days, then declined to basal levels at 5 days after injury. Immunohistochemistry further confirmed that GRP94 immunoactivity was expressed at low levels in gray matter and white matter in normal condition and increased after SCI. Double immunofluorescence staining showed that GRP94 was co-expressed with NeuN (neuronal marker), and GFAP (astroglial marker). In addition, caspase-12, caspase-3 and phospho-c-Jun NH2-kinase (p-JNK) levels increased at 6h, peaked at 1day, and then gradually reduced to normal levels for 2 weeks after SCI by western blot analysis. Co-localization of GRP94/caspase-12 and GRP94/p-JNK was detected in neurons and glial cells. Taken together, these data suggest GRP94 involvement in the injury response of the adult spinal cord of the rats., (Copyright © 2011. Published by Elsevier B.V.)

The present research was aimed to explore the biocompatibility of IKVAV self-assembling peptide nanofiber scaffold with olfactory ensheathing cells (OECs) of rats. The OECs were seeded onto the surface of coverslips covered with IKVAV self-assembling peptide nanofiber scaffold hydrogel (2D culture system), and implanted within IKVAV self-assembling peptide nanofiber scaffold hydrogel (3D culture system), respectively. The adhesion, viability of OECs were observed with inverted microscope. Then the characteristics for survival and adhesion of cells by image processing were observed, and statistical analysis on the number of S-100 positive cell, the area of the cell bodies and the perimeter of the cell and MTT method were carried out. It was found that the OECs could survive and migrate in IKVAV self-assembling peptide nanofiber scaffold. The result of the cell MTT exam, of the shape and quantity of cells had no significant difference compared to those of the OECs cultured with poly-L-lysine (PLL). It has been proved that IKVAV self-assembling peptide nanofiber scaffold has good biocompatibility with rat OECs.

This study examined bacterial growth and type on biofilm-controlling dental unit waterline (DUWL) tubing (T) and control manufacturer's tubing (C) in a laboratory DUWL model using ultrapure source water that was cycled through the lines. Sections of tubing lines were detached and examined for biofilm growth using SEM imaging at six sampling periods. Bacteria from inside surfaces of T and C, source unit, and reservoir were cultured and enumerated. At six months, organisms were molecularly identified from the alignment matches obtained from the top three BLAST searches for the 16S region. There was a 1-3 log increase in organism growth in a clean, nonsterile reservoir within an hour. Biofilm was established on the inside surfaces of C within three weeks, but not on T. Proteobacteria, and Sphingomonas spp. were identified in the source reservoir and C line, and a variation of the genera was found in T line.

Objective: A study was conducted to test the biofilm-controlling functions of N-halamine tubing over an eight-month period., Methods: A laboratory system, simulating a teaching dental clinic, was used to test rechargeable N-halamine tubing (T) compared to an untreated control (C) using the unit manufacturer's tubing. For the long-term study, a recharged tubing (RC) treated with bleach was used to compare with the test (T) and the control (C) tubing. Source tap water was cycled through the lines at 1.4 mL/minute, five minutes on and 25 minutes off, eight hours/day, five days/week. Every three weeks, samples of effluent, recovered adherent bacteria from inside tubing surfaces, and SEM images were examined for bacterial and biofilm growth. After sampling, a recharging solution of chlorine bleach (1 : 10 dilution) was run through T and RC lines, left overnight, and rinsed out the next morning. One-way ANOVAs and Spearman correlations were performed to detect significant differences for T, RC, and C, and determine significance with time period and source water, respectively., Results: Mean log CFU/mL for C effluent > T (p = 0.028), and C tubing > T (p = 0.035). Spearman correlations were significant between effluent and source water level for T (rho = 0.817), and T tubing (0.750); between RC tubing and source water level (rho = 0.836), and time (rho = 0.745); and between C and time (rho = 0.873). SEM imaging confirmed the presence of biofilm inside RC and C, but not inside T., Conclusion: N-halamine tubing completely inhibited biofilm formation without negatively affecting the physical properties of the effluent water. Further research on N-halamine tubing using a pure water source is recommended, as T effluent bacterial levels reflected the source tap water quality and proliferation of planktonic bacteria with no biofilm activity.

N-chloro-2,2,6,6-tetramethyl-4-piperidinol laurate (Cl-TMPL) was prepared by reacting 2,2,6,6-tetramethyl-4-piperidinol hydrochloride (TMP·HCl) with lauroyl chloride, followed by chlorination with sodium dichloroisocyanurate. The chemical structure of Cl-TMPL was characterized with FT-IR, NMR, DSC, and TGA analyses. The antimicrobial performance of Cl-TMPL against Gram-positive and Gram-negative bacteria was compared with 1-chloro-3-dodecyl-5,5-dimethylhydantoin (Cl-DDMH), a amide N-halamine, and chloro-2,4-diamino-6-dodecyl-1,3,5-triazine (Cl-DADT), a melamine (imino) N-halamine. The three classes of N-halamines were used as additives for polyurethane (PU). Visible light transparency data indicated that up to 6% of Cl-DDMH or Cl-DADT could be compatibly mixed with PU, but Cl-TMPL had low compatibility with PU at higher than 2% of Cl-TMPL. With the same additive content, Cl-DDMH and Cl-DADT provided more powerful antimicrobial and biofilm-controlling effects than Cl-TMPL. In stability studies, however, PU samples with Cl-TMPL released the lowest amount of active chlorine into the immersing solution, suggesting the highest stability of the antimicrobial and biofilm-controlling efficacy.

Chitosan, a natural cationic polymer, strongly binds anionic antibiotics (e.g., rifampin) through the formation of ionic complexes. The new system shows sustained rifampin release, leading to potent antimicrobial and biofilm-controlling functions against gram-positive bacteria including Staphylococcus epidermidis and Staphylococcus aureus, which are responsible for a wide range of medical device-related infections, for longer than 30 days. Moreover, the released drugs can be recharged to further extend antimicrobial durations. Drug release mechanisms and potential applications of the new system are also discussed., (Copyright 2008 Wiley Periodicals, Inc.)

In this study, the surfaces of polypropylene tubing were hydroxylated with potassium persulfate. The resultant tubing surfaces were grafted with methacrylamide (MAA) using ceric(IV) ammonium nitrate as an initiator. Upon chlorination treatment with diluted chlorine bleach, some of the amide groups in the grafted MAA side chains were transformed into stable acyclic N-halamines. The reactions were confirmed with attenuated total reflectance infrared, X-ray photoelectron spectra, and iodimetric titration. The resultant tubing was challenged with Pseudomonas aeruginosa (P. aeruginosa) in a continuous flowing model. Bacteria culturing and scanning electron microscope studies showed that the chlorinated MAA-grafted tubing could provide potent and rechargeable biofilm-controlling functions against the test microorganisms., ((c) 2007 Wiley Periodicals, Inc.)

In this study, 2-amino-4-chloro-6-hydroxy-s-triazine (ACHT) was synthesized through controlled hydrolysis of 2-amino-4,6-dichloro-s-triazine (ADCT). A simple pad-dry-cure approach was employed to immobilize ACHT onto cellulosic fibrous materials. After treatment with diluted chlorine bleach, the covalently bound ACHT moieties were transformed into chloromelamines. The structures of the samples were fully characterized with NMR, UV/VIS, DSC, TG, iodometric titration and elemental analyses. The chloromelamine-based fibrous materials provided potent, durable, and rechargeable biocidal functions against bacteria (including multi-drug resistant species), yeasts, viruses, and bacterial spores. SEM studies demonstrated that the new fibrous materials could effectively prevent the formation of biofilms, and controlled release investigations in vitro suggested that the biocidal activities were bioresponsive. Biocidal mechanisms of the chloromelamine-based fibrous materials were further discussed.

An N-halamine-based polymeric additive, poly[(6-morpholino-s-triazine-2,4-diyl)-N-chloro-[2,2,6,6-tetramethyl-4-piperidyl)imino]-hexamethylene[(2,2,6,6-4-piperidyl) imino]] (APA-1), was prepared in this study. The chemical structure of APA-1 was confirmed by iodometric titration, (1)H NMR, and (13)C NMR analysis. APA-1 was incorporated into polyurethane through solvent casting. The resultant materials demonstrated potent antimicrobial functions against Staphylococcus epidermidis. Zone of inhibition studies showed that APA-1 inactivated the bacteria through contact, without leaching out of the polyurethane samples. In vitro studies illustrated that APA-1-containing polyurethane could prevent the colonization and biofilm formation of S. epidermidis for at least 28 days.