Cryomilling-induced solid dispersion of poor glass forming/poorly water-soluble mefenamic acid with polyvinylpyrrolidone K12.

The effect of mechanical impact on the polymorphic transformation of mefenamic acid (MFA) and the formation of a solid dispersion of mefenamic acid, a poor glass forming/poorly-water soluble compound, with polyvinylpyrrolidone (PVP) K12 was investigated. The implication of solid dispersion formation on solubility enhancement of MFA, prepared by cryomilling, was investigated. Solid state characterization was conducted using powder X-ray diffraction (PXRD) and Fourier-transform infrared (FTIR) spectroscopy combined with crystal structure analysis. Apparent solubility of the mixtures in pH 7.4 buffer was measured. A calculation to compare the powder patterns and FTIR spectra of solid dispersions with the corresponding physical mixtures was conducted. Solid state characterization showed that (1) MFA I transformed to MFA II when pure MFA I was cryogenically milled (CM); and (2) MFA forms a solid dispersion when MFA was cryogenically milled with PVP K12. FTIR spectral analysis showed that hydrogen bonding facilitated by mechanical impact played a major role in forming solid dispersions. The apparent solubility of MFA was significantly improved by making a solid dispersion with PVP K12 via cryomilling. This study highlights the importance of cryomilling with a good hydrogen bond forming excipient as a technique to prepare solid dispersion, especially when a compound shows a poor glass forming ability and therefore, is not easy to form amorphous forms by conventional method.

Biosorption of sub-micron-sized polystyrene microplastics using bacterial biofilms.

Microplastics are becoming a global concern because they pose potential ecological and toxicological risks to organisms. Thus, removing microplastics from aquatic environments is important. In this study, we evaluated the capability of bacterial biofilms as a biological source for the biosorptive removal of sub-micron-sized polystyrene (PS) microplastics. Three bacterial strains-specifically, Pseudomonas aeruginosa, Bacillus subtilis, and Acinetobacter sp.-were used to form biofilms, and each biofilm was tested in batch experiments for the removal of sub-micron-sized PS microplastics. The Acinetobacter sp. biofilm demonstrated excellent removal performance against 430 nm-PS microplastics than other bacterial biofilms and showed a removal capacity of 715.5 mg/g upon treatment with the PS microplastics for 20 min, thus it employed further adsorption experiments. The biosorption of 430 nm-PS microplastics onto the Acinetobacter sp. biofilm was well explained by the pseudo-second-order kinetics and Freundlich isotherm models. Fourier transform infrared analysis indicated that biosorption of 430 nm-PS microplastics onto the Acinetobacter sp. biofilm involved chemisorption. Three environmental parameters-temperature, pH, and coexisting ions-marginally affected the biosorption of 430 nm-PS microplastics onto Acinetobacter sp. biofilm. However, the biosorption capability of Acinetobacter sp. biofilm was diminished when the 430 nm-PS microplastics were incubated in environmental freshwaters for 7 d.

Adsorption behavior of polyamide microplastics as a vector of the cyanotoxin microcystin-LR in environmental freshwaters.

Microplastics are ubiquitous environmental contaminants, and concern about microplastics functioning as vectors for coexisting environmental contaminants has been increasing. In this study, we evaluated the potential of microplastics as a vector for microcystins (MCs) in an aquatic environment. Six microplastics-polyvinylidene chloride, polystyrene, polyamide-6 (PA-6), polyvinyl chloride, poly(ethylene terephthalate), and polyethylene-were used in the experiments, and the PA-6 microplastics showed strong affinity toward the cyanotoxin microcystin-leucine arginine (MC-LR) with an adsorption efficiency of 89.5 ± 0.1 %. The adsorption of MC-LR onto PA-6 microplastics was well described by the pseudo-first-order kinetics and Langmuir isotherm models, and the adsorption was considered to be driven mainly by polar-polar interactions. The maximum adsorption capacity (qm) of MC-LR onto PA-6 microplastics was estimated to be 85.64-129.05 µg per g of PA-6 microplastics. Coexisting ions of NaCl, MgSO4, KH2PO4, CaCO3, and Na2HPO4 marginally affected the adsorption of MC-LR onto the PA-6 microplastics. However, water-quality parameters of conductivity and total-nitrogen content in environmental freshwaters influenced the adsorption of MC-LR onto PA-6 microplastics. The adsorption capability of PA-6 microplastics was evaluated using extracellular MCs (i.e., MC-LR, MC-YR, MC-RR, and total MCs) released from Microcystis aeruginosa cells during their growth.

Maternal exposure to polystyrene microplastics impairs social behavior in mouse offspring with a potential neurotoxicity.

As plastic production has been increasing steadily, environmental pollution resulting from microplastics (MPs) continues to draw considerable attention of the researchers. Several studies have reported that MPs are risk factors for various cellular and systemic dysfunctions. However, the effects of chronic MP exposure from the embryonic stage to adulthood on mouse brain remain unclear. Accordingly, determining the impacts of maternal exposure to MPs on mouse offspring was the main goal of this study. To this end, single cells of primary cortical neurons were isolated from mouse embryos. Subsequently, the cells were exposed to 2 µm polystyrene microplastics (PS-MPs), which resulted in a notable reduction in dendritic length, and PS-MPs cannot pass through the cellular membrane of neurons. Moreover, exposure to PS-MPs caused the proliferation increase and apoptosis in primary cortical neuronal cells. We then evaluated the neurotoxicity associated with chronic PS-MP exposure from the embryonic stage to adulthood in C57BL/6 J mouse offspring. PS-MPs were found to accumulate in the digestive and excretory organs of the offspring but not in the brain tissue. However, offspring exposed to PS-MPs exhibited no differences in the levels of expression of genes related to brain cell markers or synaptic organization. Nevertheless, PS-MP-exposed mice exhibited impaired social novelty preferences; however, no changes were observed in the emotional, compulsive, or cognitive behaviors. Taken together, these results demonstrate the potential neurotoxic effects of chronic exposure to PS-MPs in mouse offspring.

Exposure to polystyrene particles causes anxiety-, depression-like behavior and abnormal social behavior in mice.

In the era of plastic use, organisms are constantly exposed to polystyrene particles (PS-Ps). PS-Ps accumulated in living organisms exert negative effects on the body, although studies evaluating their effects on brain development are scarce. In this study, the effects of PS-Ps on nervous system development were investigated using cultured primary cortical neurons and mice exposed to PS-Ps at different stages of brain development. The gene expression associated with brain development was downregulated in embryonic brains following PS-Ps exposure, and Gabra2 expression decreased in the embryonic and adult mice exposed to PS-Ps. Additionally, offspring of PS-Ps-treated dams exhibited signs of anxiety- and depression-like behavior, and abnormal social behavior. We propose that PS-Ps accumulation in the brain disrupts brain development and behavior in mice. This study provides novel information regarding PS-Ps toxicity and its harmful effects on neural development and behavior in mammals.

Ionically bridged dexamethasone sodium phosphate-zinc-PLGA nanocomplex in alginate microgel for the local treatment of ulcerative colitis.

Colon-targeted oral drug delivery systems comprising nanoparticles and microparticles have emerged as promising tools for the treatment of ulcerative colitis (UC) because they minimize side effects and maximize the local drug concentration. Dexamethasone sodium phosphate (DSP) is a potent anti-inflammatory glucocorticoid used for the treatment of UC. However, it remains a rather short-term treatment option owing to its side effects. In the present study, we developed the alginate gel encapsulating ionically bridged DSP-zinc-poly(lactic-co-glycolic acid) (PLGA) nanocomplex (DZP-NCs-in-microgel) for the oral local treatment of UC. The successful encapsulation of DSP-zinc-PLGA nanocomplex (DZP-NCs) in alginate microgel was confirmed by SEM imaging. The prepared gel released DZP-NCs in the stimulated intestinal fluid and dampened the release of DSP in the upper gastrointestinal tract. Furthermore, DZP-NCs-in-microgel alleviated colonic inflammation in a mouse model of dextran sodium sulfate-induced colitis by relieving clinical symptoms and histological marks. Our results suggest a novel approach for the oral colon-targeted delivery of dexamethasone sodium phosphate for the treatment of UC.

Adsorptive removal of micron-sized polystyrene particles using magnetic iron oxide nanoparticles.

Microplastics are able to pass through many filtration systems due to their small sizes, making it difficult to remove them from, for example, water. In this study, we evaluated the ability of using magnetic iron oxide (Fe3O4) nanoparticles to achieve the adsorptive removal of micron-sized polystyrene (microPS) particles. Application of a magnet for 3 min to an aqueous sample of microPS particles mixed with iron oxide nanoparticles for 1 min was able to effectively remove the microPS particles from the water. Transmission electron microscopy images of such samples showed the formation of Fe3O4-PS complexes due to the adsorption of PS particles onto iron oxide nanoparticles. This adsorption followed the pseudo-first order kinetic and Langmuir isotherm model. Hydrophobic interactions were concluded from our experiments to be the main interactions involved in the aggregation of iron oxide with PS particles. Ions present in an environmental freshwater sample inhibited the ability of iron oxide particles to become adsorbed PS particles, but the adsorption performance was improved by increasing the amount of iron oxide particles. The iron oxide particles could be recovered from the Fe3O4-PS complexes by desorption process. Our study showed the potential advantages of iron oxide particles for removing environmental pollutants of microplastics via highly efficient and environmental-friendly procedure.

Removable coatings: Thermal stability and decontamination of steel surfaces from 241Am.

This study presents a comparative analysis of several commercial removable materials for radioactive decontamination of steel surfaces using 241Am as representative radionuclide. The selection criteria of removable coatings for this study included a history of application, commercial availability, easy handling conditions and different composition and formulation. Carbon steel and stainless steel coupons were utilized as common industrial materials, and the experimental series were expanded to include the rusting treatment of these surfaces as it is common for decommissioned nuclear facilities. Radionuclide 241Am was deposited on the coupon surfaces and used to evaluate decontamination efficiency of the removable coatings, which were pre-screened for the ease of application and removal from the surface. Selected coatings were characterized with Fourier-transform infrared spectroscopy and thermogravimetric analysis, decontamination efficiencies for different types of steel surfaces, and potential enhancement of the removal efficiencies of the select removable coatings via amendment with EDTA. Across all the coatings, decontamination efficiencies for stainless steel (both pristine and with oxidizing treatment) were higher than for pristine carbon steel, which in turn were higher than for rusted carbon steel. Amendment with EDTA improved removal efficiency of a removable coating. CC Strip coating exhibited easy handling and high decontamination efficiency, (up to 97% when EDTA-amended), but its drying time was the longest, and thermal analysis indicated higher release of energy during thermal decomposition compared to the other coatings. Hydrogel-based DeconGel coating, even though not the easiest in handling among the rest of materials, exhibited high decontamination efficiency, efficient drying at the ambient temperature leading to the loss of about 80 wt% due to solvent evaporation, and extremely low heat released during thermal decomposition; therefore, it is considered a preferable choice for the considered factors.

Interactions between bacteria and nano (micro)-sized polystyrene particles by bacterial responses and microscopy.

Microorganisms play an important role in biogeochemical cycles, and are inevitably found associated with plastic debris. The interplay between microbes and plastics may change the characteristics of certain plastics over time and drive the environmental fate of plastics. In this study, we evaluated interactions of bacteria with nano- and microplastics. Here, polystyrene (PS) polymer particles of various diameters, specifically 60, 220, 430, 700, 1040, 1700, and 2260 nm, were used as the plastics. Escherichia coli (E. coli, gram-negative) and Bacillus sp. (gram-positive) were chosen as model bacteria. The effects of nano- and microPS particles on E. coli and Bacillus sp. cells were investigated by measuring the growth and viability of the cells in laboratory-scale flasks and their generation of reactive oxygen species (ROS) upon their exposure to these particles of 100 mg/L. The particles inhibited the growth and viability of both types of bacterial cells, but their inhibitory effects varied depending on the diameter of PS particle. The 60-nm-diameter PS particles were visually observed to enter the cells as well as accumulate on their surfaces and enhanced ROS generation of the cells. Unexpectedly, the 1040-nm-diameter PS particles, similar in size to the bacterial cells, inhibited the growth of both E. coli and Bacillus sp. cells the most. The E. coli and Bacillus sp. cells formed microPS-biofilm complex by secreting an extracellular polymeric substance (EPS) in response to their exposure to the ∼ 1-µm-diameter PS particles. A positive correlation between relative ROS levels and specific growth rates of the E. coli cells were observed with a Pearson correlation coefficient r value of 0.676 (p < 0.05).

Clinical Anti-aging Efficacy of Propolis Polymeric Nanoparticles Prepared by a Temperature-induced Phase Transition Method.

BACKGROUND: Collagen forms a dermal matrix in the skin. Biosynthesis and decomposition of collagen are the major processes in skin aging. Propolis is rich in flavonoids and phenolic compounds, which are known to be effective in preventing skin aging, including the enhancement of fibroblast proliferation, activation, and growth capacity. OBJECTIVES: The aim of this study was to develop a poorly soluble propolis extract as an active ingredient in cosmetic products for anti-aging efficacy. METHODS & RESULTS: Polymeric nanoparticles containing propolis extract, polyethylene glycol 400, and poloxamer 407 were prepared via a temperature-induced phase transition method. The particle size of the polymeric nanoparticles was approximately 20.75 nm. The results of an in vitro procollagen type I carboxy-terminal peptide assay and a matrix metalloproteinase-1 inhibition assay showed that the polymeric nanoparticles increased collagen production by 19.81%-24.59% compared to blank (p < 0.05), and significantly reduced intracellular collagenase activity by 7.46%-31.52% compared to blank (p < 0.05). In a clinical trial, polymeric nanoparticles in a cosmetic formulation were applied around the eyes of 24 female subjects for 8 weeks. Five skin parameters were significantly improved after the application of the test ampoule. Visual evaluation using the Global Photo Damage Score showed a significant reduction in wrinkles after the application of the test ampoules (p < 0.001). CONCLUSIONS: This study outlines the development of stable polymeric nanoparticles containing poorly soluble propolis in a cosmetic formulation, and its efficacy in wrinkle improvement. The developed polymeric nanoparticles were effective for alleviating wrinkles and can be used for pharmaceutical applications that utilize propolis as antiseptic, anti-inflammatory, antimycotic, antifungal, antibacterial, antiulcer, anticancer, and immunomodulatory agents.

Enhancing solubility and bioavailability of coenzyme Q10: formulation of solid dispersions using Soluplus® as a carrier.

Improving the aqueous solubility of poorly soluble compounds have been a major issue in the pharmaceutical industry. In the present study, binary amorphous solid dispersions (SDs) of Coenzyme Q10 (CoQ10), a biopharmaceutics classification system (BCS) II compound and Soluplus® were prepared to enhance the solubility and pharmacokinetic properties compared to crystalline CoQ10. SDs were prepared with different ratios of CoQ10 and Soluplus® (1:3, 1:5, and 1:7) using spray drying technology, and the physicochemical properties of the SDs were evaluated. X-ray powder diffraction, differential scanning calorimetry, and scanning electron microscopy suggested the conversion of the crystalline form of CoQ10 to a binary amorphous system in the SDs. Fourier transform infrared spectroscopy revealed no potential interactions between CoQ10 and Soluplus®. The solubility of the optimal SD formulation (SD 1:7) was approximately 9000-fold higher than that of crystalline CoQ10, and the increment was Soluplus® concentration dependent. As a result, optimized SD 1:7 also showed significantly enhanced dissolution rate where maximum drug release was observed within 30 min in two different dissolution media. Moreover, in contrast to crystalline CoQ10, CoQ10 SDs showed improved pharmacokinetic parameters. Thus, the SD 1:7 formulation is expected to improve biopharmaceutical properties and therapeutic efficacy of CoQ10.

Bacteria-Adhesive Nitric Oxide-Releasing Graphene Oxide Nanoparticles for MRPA-Infected Wound Healing Therapy.

A bacteria-infected wound can lead to being life-threatening and raises a great economic burden on the patient. Here, we developed polyethylenimine 1.8k (PEI1.8k) surface modified NO-releasing polyethylenimine 25k (PEI25k)-functionalized graphene oxide (GO) nanoparticles (GO-PEI25k/NO-PEI1.8k NPs) for enhanced antibacterial activity and infected wound healing via binding to the bacterial surface. In vitro antibacterial activity and in vivo wound healing efficacy in an infected wound model were evaluated compared with NO-releasing NPs (GO-PEI25k/NO NPs). Surface modification with PEI1.8k can enhance the ability of nanoparticles to adhere to bacteria. GO-PEI25k/NO-PEI1.8k NPs released NO in a sustained manner for 48 h and exhibited the highest bactericidal activity (99.99% killing) against methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa (MRPA) without cytotoxicity to L929 mouse fibroblast cells at 0.1 mg/mL. In the MRPA-infected wound model, GO-PEI25k/NO-PEI1.8k NPs showed 87% wound size reduction while GO-PEI25k/NO NPs showed 23% wound size reduction at 9 days postinjury. Masson trichrome and hematoxylin and eosin staining revealed that GO-PEI25k/NO-PEI1.8k NPs enhanced re-epithelialization and collagen deposition, which are comparable to healthy mouse skin tissue. GO-PEI25k/NO-PEI1.8k NPs hold promise as effective antibacterial and wound healing agents.

Effect of switching gas inlet position on the performance of a polyurethane biofilter under transient loading for the removal of benzene, toluene and xylene mixtures.

The performance of a polyurethane (PU) biofilter was evaluated using different operating modes (unidirectional flow (UF) and flow-directional switching (FDS) operations) under transient loading conditions (intermittent and shutdown). Gas mixtures containing benzene, toluene and xylene (BTX) were employed as model gases. Quantitative real-time PCR methods were used for targeting the tmoA gene responsible for BTX degradation and estimating density of the BTX-degraders in the PU filter bed. Although the overall BTX Removal efficiencies at the outlet (50 h(-1) of space velocity) were similar between the UF and FDS biofilters, the removability of BTX in the FDS biofilter was higher than that in the UF biofilter until the 3rd sampling position (68 h(-1) of space velocity). The BTX removal potentials and tmoA gene copy numbers of the FDS biofilter remained constant, irrespective of the distances from the inlet, but those of the UF biofilter increased with increasing distance from the inlet position. These results indicate that an even distribution of BTX degraders in the FDS filter bed contributed to better BTX removal performance. After a 10 day-shutdown, the performances of the UF and SDF biofilters were rapidly restored within 1 day.

Simple screening of microplastics in bottled waters and environmental freshwaters using a novel fluorophore.

Screening of polymeric microplastic debris can help to assess the extent to which plastics contaminate the environment. We here developed an easy- and rapid-to-perform method for the screening of plastic polymers, based on a newly employed fluorophore, namely 1-pyrenebutyric acid N-hydroxysuccinimidyl ester (PBN). The PBN fluorophore was capable of staining diverse synthetic microplastic polymers within 5 min, including those displaying various particle sizes and shapes. The fluorescence intensities of the microplastics were considerably enhanced after the short-duration staining. The screening method was shown to be highly effective in the detection of polyethylene (PE), poly(ethylene terephthalate) (PET), polyvinyl chloride (PVC), polyamide-6 (PA-6), polypropylene (PP), polystyrene (PS), polycarbonate (PC), polyvinylidenechloride (PVDC), and polyurethane (PU), with a lowest analyzed particle size of 15 µm. Using our screening method, plastic contamination was investigated in commercially available bottled waters and environmental waters, specifically urban freshwaters. This study demonstrated high affinity levels of the newly proposed PBN fluorophore for a broad range of polymers and its ability to be used to discernibly identify polymer particles.

Microarray-based analysis of the lung recovery process after stainless-steel welding fume exposure in Sprague-Dawley rats.

Repeated exposure to welding fumes promotes a reversible increase in pulmonary disease risk, but the molecular mechanisms by which welding fumes induce lung injury and how the lung recovers from such insults are unclear. In the present study, pulmonary function and gene-expression profiles in the lung were analyzed by Affymetrix GeneChip microarray after 30 days of consecutive exposure to manual metal arc welding combined with stainless-steel (MMA-SS) welding fumes, and again after 30 days of recovery from MMA-SS fume exposure. In total, 577 genes were identified as being either up-regulated or down-regulated (over twofold changes, p < 0.05) in the lungs of low-dose or high-dose groups. Differentially expressed genes were classified based on a k-means clustering algorithm and biological functions and molecular networks were further analyzed using Ingenuity Pathways Analysis. Among the genes affected by exposure to or recovery from MMA-SS fumes, the transcriptional changes of 13 genes that were highly altered by treatment were confirmed by quantitative real-time PCR. Notably, Mmp12, Cd5l, Ccl7, Cxcl5, and Spp1 related to the immune response were up-regulated only in the exposure group, whereas Trem2, IgG-2a, Igh-1a, and Igh were persistently up-regulated in both the exposure and recovery groups. In addition, several genes that might play a role in the repair process of the lung were up-regulated exclusively in the recovery group. Collectively, these data may help elucidate the molecular mechanism of the recovery process of the lung after welding fume exposure.

Platform-independent models for age prediction using DNA methylation data.

Age prediction has been in the spotlight recently because it can provide an important information about the contributors of biological evidence left at crime scenes. Specifically, many researchers have actively suggested age-prediction models using DNA methylation at several CpG sites and tested the candidates using platforms such as the HumanMethylation 450 array and pyrosequencing. With DNA methylation data obtained from each platform, age prediction models were constructed using diverse statistical methods typically with multivariate linear regression. However, because each developed model is based on single-platform data, the prediction accuracy is reduced when applying DNA methylation data obtained from other platforms. In this study, bisulfite sequencing data for 95 saliva samples were generated using massively parallel sequencing (MPS) and compared with methylation SNaPshot data from the same 95 individuals. The predicted age obtained by applying MPS data to an age-prediction model built for methylation SNaPshot data differed greatly from the chronological age due to platform differences. Therefore, novel variables were introduced to indicate the platform type, and construct platform-independent age predictive models using a neural network and multivariate linear regression. The final neural network model had a mean absolute deviation (MAD) of 3.19 years between the predicted and chronological age, and the mean absolute percentage error (MAPE) was 8.89% in the test set. Similarly, the linear regression model showed 3.69 years of MAD and 10.44% of MAPE in the same test set. The platform-independent age-prediction model was made extensible to an increasing number of platforms by introducing platform variables, and the idea of platform variables can be applied to age prediction models for other body fluids.

DNA methylation of the ELOVL2, FHL2, KLF14, C1orf132/MIR29B2C, and TRIM59 genes for age prediction from blood, saliva, and buccal swab samples.

Many studies have reported age-associated DNA methylation changes and age-predictive models in various tissues and body fluids. Although age-associated DNA methylation changes can be tissue-specific, a multi-tissue age predictor that is applicable to various tissues and body fluids with considerable prediction accuracy might be valuable. In this study, DNA methylation at 5 CpG sites from the ELOVL2, FHL2, KLF14, C1orf132/MIR29B2C, and TRIM59 genes were investigated in 448 samples from blood, saliva, and buccal swabs. A multiplex methylation SNaPshot assay was developed to measure DNA methylation simultaneously at the 5 CpG sites. Among the 5 CpG sites, 3 CpG sites in the ELOVL2, KLF14 and TRIM59 genes demonstrated strong correlation between DNA methylation and age in all 3 sample types. Age prediction models built separately for each sample type using the DNA methylation values at the 5 CpG sites showed high prediction accuracy with a Mean Absolute Deviation from the chronological age (MAD) of 3.478 years in blood, 3.552 years in saliva and 4.293 years in buccal swab samples. A tissue-combined model constructed with 300 training samples including 100 samples from each blood, saliva and buccal swab samples demonstrated a very strong correlation between predicted and chronological ages (r = 0.937) and a high prediction accuracy with a MAD of 3.844 years in the 148 independent test set samples of 50 blood, 50 saliva and 48 buccal swab samples. Although more validation might be needed, the tissue-combined model's prediction accuracies in each sample type were very much similar to those obtained from each tissue-specific model. The multiplex methylation SNaPshot assay and the age prediction models in our study would be useful in forensic analysis, which frequently involves DNA from blood, saliva, and buccal swab samples.

S-Nitrosoglutathione loaded poly(lactic-co-glycolic acid) microparticles for prolonged nitric oxide release and enhanced healing of methicillin-resistant Staphylococcus aureus-infected wounds.

Methicillin-resistant Staphylococcus aureus (MRSA)-infected wounds have become a significant clinical issue worldwide. Recently, nitric oxide (NO) has emerged as a potent antibacterial agent against MRSA infections and a wound-healing enhancer. Nevertheless, clinical applications of NO have been largely restricted by its gaseous state and short half-life. In this study, our aim was to develop S-nitrosoglutathione (GSNO, an endogenous NO donor)-loaded poly(lactic-co-glycolic acid) [PLGA] microparticles (GSNO-MPs) that release NO over a prolonged period, to accelerate the healing of MRSA-infected wounds with less frequent dosing. GSNO was successfully encapsulated into PLGA microparticles by a solid-in-oil-in-water emulsion solvent evaporation method. Scanning electron microscopy and X-ray diffraction analyses confirmed the successful fabrication of GSNO-MPs. The latter released NO in a prolonged manner over 7 days and exerted a remarkable antibacterial activity against MRSA in a concentration- and time-dependent manner. Moreover, GSNO-MPs had good antibacterial efficacy and were found to accelerate wound healing in a mouse model of MRSA-infected wounds. Therefore, NO-releasing MPs devised in this study may be a promising option for the treatment of cutaneous wounds infected by drug-resistant bacteria such as MRSA.

DNA methylation-based age prediction from saliva: High age predictability by combination of 7 CpG markers.

DNA methylation is currently one of the most promising age-predictive biomarkers. Many studies have reported DNA methylation-based age predictive models, but most of these are based on DNA methylation patterns from blood. Only a few studies have examined age-predictive DNA patterns in saliva, which is one of the most frequently-encountered body fluids at crime scenes. In this study, we generated genome-wide DNA methylation profiles of saliva from 54 individuals and identified CpG markers that showed a high correlation between methylation and age. Because the age-associated marker candidates from saliva differed from those of blood, we investigated DNA methylation patterns of 6 age-associated CpG marker candidates (cg00481951, cg19671120, cg14361627, cg08928145, cg12757011, and cg07547549 of the SST, CNGA3, KLF14, TSSK6, TBR1, and SLC12A5 genes, respectively) in addition to a cell type-specific CpG marker (cg18384097 of the PTPN7 gene) in an independent set of saliva samples obtained from 226 individuals aged 18 to 65 years. Multiplex methylation SNaPshot reactions were used to generate the data. We then generated a linear regression model with age information and the methylation profile from the 113 training samples. The model exhibited a 94.5% correlation between predicted and chronological age with a mean absolute deviation (MAD) from chronological age of 3.13 years. In subsequent validation using 113 test samples, we also observed a high correlation between predicted and chronological age (Spearman's rho=0.952, MAD from chronological age=3.15years). The model composed of 7 selected CpG sites enabled age prediction in saliva with high accuracy, which will be useful in saliva analysis for investigative leads.

Temporomandibular joint synovial chondromatosis extending to the temporal bone: a report of two cases.

Synovial chondromatosis is a rare benign lesion originating from the synovial membrane. It presents as adhesive or non-adhesive intra-articular cartilaginous loose bodies. Although the causes of synovial chondromatosis have not been fully elucidated, inflammation, external injury, or excessive use of joints have been suggested as possible causes. Synovial chondromatosis has been reported to occur most frequently at large joints that bear weights, with a rare occurrence at the temporomandibular joint (TMJ). When synovial chondromatosis develops at TMJ, clinical symptoms, including pain, joint sounds, and mouth opening may common. Moreover, synovial chondromatosis rarely spreads to the mandibular condyle, glenoid cavity, or articular eminence of TMJ. The goal of this study was to discuss the methods of surgery and other possible considerations by reviewing cases of patients who underwent surgery for synovial chondromatosis that extended to the temporal bone.