Antimicrobial Activity of Smilax china L. Root Extracts against the Acne-Causing Bacterium, Cutibacterium acnes, and Its Active Compounds.

The root of Smilax china L. is used in traditional Korean medicine. We found that the Smilax china L. root extract has strong antimicrobial activity against two Cutibacterium acnes strains (KCTC 3314 and KCTC 3320). The aim of this study was to identify the beneficial properties of Smilax china L. extracts for their potential use as active ingredients in cosmetics for the treatment of human skin acne. The high-performance liquid chromatography (HPLC) and liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC/QTOF/MS) methods were used to obtain the profile of secondary metabolites from the ethyl acetate-soluble fraction of the crude extract. Agar diffusion and resazurin-based broth microdilution assays were used to evaluate antimicrobial activity and minimum inhibitory concentrations (MIC), respectively. Among the 24 metabolites, quercetin, resveratrol, and oxyresveratrol were the most potent compounds against Cutibacterium acnes. Minimum inhibitory concentrations of quercetin, resveratrol, and oxyresveratrol were 31.25, 125, and 250 µg/mL, respectively.

Fundamental Understanding of the Formation Mechanism for Graphene Quantum Dots Fabricated by Pulsed Laser Fragmentation in Liquid: Experimental and Theoretical Insight.

The pulsed laser fragmentation in liquid (PLFL) process is a promising technique for the synthesis of carbon-based functional materials. In particular, there has been considerable attention on graphene quantum dots (GQDs) derived from multiwalled carbon nanotubes (MWCNTs) by the PLFL process, owing to the low cost and rapid processing time involved. However, a fundamental deep understanding of the formation of GQDs from MWCNTs by PLFL has still not been achieved despite the high demand. In this work, a mechanism for the formation of GQDs from MWCNTs by the PLFL process is reported, through the combination of experimental and theoretical studies. Both the experimental and computational results demonstrate that the formation of GQDs strongly depends on the pulse laser energy. Both methods demonstrate that the critical energy point, where a plasma plume is generated on the surface of the MWCNTs, should be precisely maintained to produce GQDs; otherwise, an amorphous carbon structure is favorably formed from the scattered carbons.

One-step synthesis of sulfur-incorporated graphene quantum dots using pulsed laser ablation for enhancing optical properties.

To tune the electronic and optoelectronic properties of graphene quantum dots (GQDs), heteroatom doping (e.g., nitrogen (N), boron (B), and sulfur (S)) is an effective method. However, it is difficult to incorporate S into the carbon framework of GQDs because the atomic size of S is much larger than that of C atoms, compared to N and B. In this study, we report a simple and one-step method for the synthesis of sulfur-doped GQDs (S-GQDs) via the pulsed laser ablation in liquid (PLAL) process. The as-prepared S-GQDs exhibited enhanced fluorescence quantum yields (0.8% → 3.89%) with a huge improved absorption band in ultraviolet (UV) region (200 ∼ 400 nm) and excellent photo stability under the UV radiation at 360 nm. In addition, XPS results revealed that the PLAL process can effectively facilitate the incorporation of S into the carbon framework compared to those produced by the chemical exfoliation method (e.g., hydrothermal method). And also, the mechanisms related with the optical properties of S-GQDs was investigated by time-resolved photoluminescence (TRPL) spectroscopy. We believe that the PLAL process proposed in this study will serve as a simple and one-step route for designing S-GQDs and opens up to opportunities for their potential applications.

Escherichia coli ß-galactosidase-catalyzed synthesis of 2-phenoxyethanol galactoside and its characterization.

We synthesized 2-phenoxyethanol galactoside (PE-Gal) from 2-phenoxyethanol (PE), in which Escherichia coli ß-gal (as E. coli cells) and lactose were added in the reaction mixture for galactosylation. About 40 mM PE-Gal was maximally synthesized from about 80 mM PE at 24 h as about 50% conversion yield. After purifying PE-Gal, the structure of PE-Gal was identified using LC-MS, (1)H NMR, and (13)C NMR analyses. In addition, it was observed that the water solubility of PE-Gal was increased by galactosylation of PE. The MICs of PE and PE-Gal against Gram-negative and Gram-positive bacteria were fairly similar with each other (23.3-61.3 mM as the average value). PE-Gal was noticeably less cytotoxic against HACAT cells, in particular a remarkable difference in cell viability was observed at concentrations of 20-60 mM PE or PE-Gal. Finally, we accomplished the synthesis of less toxic PE-Gal, compared with PE, using ß-gal-containing E. coli cells without changing in the MICs against microorganisms. In the future, PE-Gal will be applicable as a substitute for PE as a less toxic preservative for the cosmetic, pharmaceutical, and food industries.

Evaluating the performance of thermoplastic 3D bolus used in radiation therapy.

A 3D-printed bolus is being developed to deliver accurate doses to superficial cancers. In this study, flexible thermoplastic filaments, specifically PLA, TPU, PETG, and HIPS, were fabricated into boluses and then compared to commercial bolus for the variation of the dose elevation region of photon beams. The experimental results indicate that the maximum dose depth is similar, and the consistent trend of the percentage depth dose confirms the potential usage as a build-up bolus.

Stable isolation of phycocyanin from Spirulina platensis associated with high-pressure extraction process.

A method for stably purifying a functional dye, phycocyanin from Spirulina platensis was developed by a hexane extraction process combined with high pressure. This was necessary because this dye is known to be very unstable during normal extraction processes. The purification yield of this method was estimated as 10.2%, whose value is 3%-5% higher than is the case from another conventional separation method using phosphate buffer. The isolated phycocyanin from this process also showed the highest purity of 0.909 based on absorbance of 2.104 at 280 nm and 1.912 at 620 nm. Two subunits of phycocyanin namely α-phycocyanin (18.4 kDa) and ß-phycocyanin (21.3 kDa) were found to remain from the original mixtures after being extracted, based on SDS-PAGE analysis, clearly demonstrating that this process can stably extract phycocyanin and is not affected by extraction solvent, temperature, etc. The stability of the extracted phycocyanin was also confirmed by comparing its DPPH (α,α-diphenyl-ß-picrylhydrazyl) scavenging activity, showing 83% removal of oxygen free radicals. This activity was about 15% higher than that of commercially available standard phycocyanin, which implies that the combined extraction method can yield relatively intact chromoprotein through absence of degradation. The results were achieved because the low temperature and high pressure extraction effectively disrupted the cell membrane of Spirulina platensis and degraded less the polypeptide subunits of phycocyanin (which is a temperature/pH-sensitive chromoprotein) as well as increasing the extraction yield.

Complete genome sequence of strain s85, a novel member of the family Flavobacteriaceae.

An agar-degrading marine bacterium identified as a novel member of the family Flavobacteriaceae (strain S85) was isolated from seawater in Micronesia. The sequenced strain S85 genome is composed of 3,384,629 bp in a circular chromosome, which includes 2,883 complete open reading frames.

Ultraviolet A regulates adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells via up-regulation of Kruppel-like factor 2.

Adipocyte dysfunction is strongly associated with the development of obesity, which is a major risk factor for many disorders, including diabetes, hypertension, and heart disease. This study shows that ultraviolet A (UVA) inhibits adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells and its action mechanisms. The mRNA levels of peroxidase proliferator-activated receptor (PPAR) γ and CCAAT/enhancer-binding protein α (C/EBPα), but not CCAAT/enhancer-binding protein ((C/EBP) ß and δ, were reduced by UVA. Moreover, the mRNA levels of PPAR γ target genes (lipoprotein lipase (LPL), CD36, adipocyte protein (aP2), and liver X receptor α (LXR)) were down-regulated by UVA. Additionally, attempts to elucidate a possible mechanism underlying the UVA-mediated effects revealed that UVA induced migration inhibitory factor (MIF) gene expression, and this was mediated through activation of AP-1 (especially JNK and p42/44 MAPK) and nuclear factor-κB. In addition, reduced adipogenesis by UVA was recovered upon the treatment with anti-MIF antibodies. AMP-activated protein kinase phosphorylation and up-regulation of Kruppel-like factor 2 (KLF2) were induced by UVA. Taken together, these findings suggest that the inhibition of adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells by UVA occurs primarily through the reduced expression of PPAR γ, which is mediated by up-regulation of KLF2 via the activation of MIF-AMP-activated protein kinase signaling.

Platelet reactivity in patients with chronic kidney disease receiving adjunctive cilostazol compared with a high-maintenance dose of clopidogrel: results of the effect of platelet inhibition according to clopidogrel dose in patients with chronic kidney disease (PIANO-2 CKD) randomized study.

BACKGROUND: Chronic kidney disease (CKD) is a factor of low response to clopidogrel. We sought to assess the functional impact of cilostazol in CKD patients with undergoing hemodialysis. METHODS: Seventy-four patients with CKD undergoing hemodialysis and percutaneous coronary intervention were enrolled. Patients were randomly assigned to receive clopidogrel (75 mg/d [group 1, n = 24]), high-maintenance dose of clopidogrel (150 mg/d [group 2, n = 25]), or clopidogrel (75 mg/d) with cilostazol (200 mg/d [group 3, n = 25]) for 14 days. Another 50 patients with normal renal function undergoing percutaneous coronary intervention were treated with 75 mg of clopidogrel and served as the control group. Platelet function was evaluated before and after antiplatelet therapy with light transmittance aggregometry and with VerifyNow P2Y12 assay (Accumetrics, San Diego, CA). Platelet activation markers (soluble CD40 ligand and soluble P-selectin) were also assessed. RESULTS: The baseline platelet function measurements were similar in the 3 groups of patients; however, the CKD groups had significantly higher platelet aggregation activity compared with the control groups. The rate of high on-treatment platelet reactivity was significantly lower in group 3 than in groups 1 and 2 (10% vs 43% vs 32%, respectively; P < .05). After 14 days of antiplatelet therapy, the changes in plasma soluble CD40 ligand and soluble P-selectin levels were significantly higher in group 3 compared with groups 1 and 2 (P < .01); however, there were no significant differences in platelet function and activation markers between groups 1 and 2. CONCLUSIONS: Adjunctive cilostazol improves platelet inhibition compared with 75 or 150 mg of clopidogrel in CKD patients undergoing hemodialysis.

A facile and efficient method for the synthesis of crystalline tetrahydro-ß-carbolines via the Pictet-Spengler reaction in water.

A facile and efficient synthesis of tetrahydro-ß-carbolines (tryptolines) in one step from tryptamine and aldehydes, in an environmentally friendly water solvent, has been investigated. This convenient and clean synthesis of various tryptolines was facilitated by L-tartaric acid, a natural compound, to obtain the desired products as clear crystals. Among the four crystalline products, the most substituted tryptoline 2 showed the best inhibitory activity against EJ cells and the least cytotoxicity, with an LC50 value of 1.49 mg/mL, against brine shrimp larvae.

Sanguinarine induces apoptosis in A549 human lung cancer cells primarily via cellular glutathione depletion.

Sanguinarine is a plant-derived benzophenanthridine alkaloid and has been shown to possess anti-tumor activities against various cancer cells. In this study, we investigated whether sanguinarine induces apoptosis in A549 human lung cancer cells. Treatment of A549 cells with sanguinarine induced apoptosis in a dose- and time-dependent manner. Treatment with sanguinarine led to activation of caspases and MAPKs as well as increased MKP-1 expression. Importantly, pretreatment with z-VAD-fmk, a pan caspase inhibitor suppressed the sanguinarine-induced apoptosis in A549 cells. Moreover, pretreatment with NAC, a sulfhydryl group-containing reducing agent strongly suppressed the apoptotic response and caspase activation to sanguinarine. However, the sanguinarine-mediated cytotoxicity in A549 cells was not protected by pharmacological inhibition of MAPKs or MKP-1 siRNA-mediated knockdown of MKP-1. These results collectively suggest that sanguinarine induces apoptosis in A549 cells through cellular glutathione depletion and the subsequent caspase activation.

Measurement of ethanol concentration using solvent extraction and dichromate oxidation and its application to bioethanol production process.

A method for measuring the ethanol concentration in a yeast culture broth was developed using both microtubes and a 96-deepwell microplate. The strategy involved first the solvent extraction of ethanol from the yeast culture broth and measurements of the ethanol concentration using the dichromate oxidation method. Particular focus was made on selecting the extraction solvent as well as determining the measurable range of ethanol concentrations using this solvent extraction-dichromate oxidation method. This method was developed as an assay format in 2.0-ml microtubes and 1.2-ml 96-deepwell microplates, and the ethanol concentration in the batch cultures and fed-batch fermentations was measured. Tri-n-butyl phosphate [non-alcoholic solvent, density = 0.9727, solubility in water = 0.028% (w/v)] was used for solvent extraction when measuring the ethanol concentration from the yeast culture broth. The maximum detectable ethanol concentration was 8% (v/v) when 10 g potassium dichromate in 100 ml of 5 M sulfuric acid was used. The concentrations determined from the solvent extraction-dichromate oxidation methods were remarkably similar to those of gas chromatography in which samples were prepared from seven experiments, such as four batch cultures and three fed-batch fermentations.

Inhibitory effects of the aqueous extract of Magnolia officinalis on the responses of human urinary bladder cancer 5637 cells in vitro and mouse urinary bladder tumors induced by N-Butyl-N-(4-hydroxybutyl) nitrosamine in vivo.

This study investigated the anticancer activity of Magnolia officinalis on urinary bladder cancer in vitro and in vivo, and elucidated the mechanism of its activity. An aqueous extract of M. officinalis inhibited cell viability and DNA synthesis in cultured human urinary bladder cancer 5637 cells. Inhibition of proliferation was the result of apoptotic induction, because FACS analyses of 5637 cells treated with M. officinalis showed a sub-G1 phase accumulation. M. officinalis extract also increased cytoplasmic DNA-histone complex dose-dependently. These inhibitory effects were associated with the upregulation of proapoptotic molecules Bax, cytochrome c and caspase 3. Treatment of 5637 cells with M. officinalis extract suppressed the expression of matrix metalloproteinase 2 (MMP-2) and MMP-9, as revealed by zymographic and immunoblot analyses. When M. officinalis extract was given to mice simultaneously with the carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine, which induces urinary bladder tumors, the size of the induced tumors was smaller. Finally, histological data indicated that the histological grade of carcinoma and the depth of invasion were dramatically decreased by treatment with M. officinalis extract in mice with N-butyl-N-(4-hydroxybutyl) nitrosamine-induced urinary bladder tumors. In conclusion, the findings showed that M. officinalis extract exhibited potential chemopreventive activity against urinary bladder tumor in vitro and in vivo.

Optimal Design of Patient-Specific Total Knee Arthroplasty for Improvement in Wear Performance.

Life expectancy is on the rise and, concurrently, the demand for total knee arthroplasty (TKA), which lasts a lifetime, is increasing. To meet this demand, improved TKA designs have been introduced. Recent advances in radiography and manufacturing techniques have enabled the production of patient-specific TKA. Nevertheless, concerns regarding the wear performance, which limit the lifespan of TKA, remain to be addressed. This study aims at reducing the wear in patient-specific TKA using design optimization and parametric three-dimensional (3D) finite-element (FE) modelling. The femoral component design was implemented in a patient-specific manner, whereas the tibial insert conformity remained to be determined by design variables. The gait cycle loading condition was applied, and the optimized model was validated by the results obtained from the experimental wear tests. The wear predictions were iterated for five million gait cycles using the computational model with force-controlled input. Similar patterns for internal/external rotation and anterior/posterior translation were observed in both initial and optimal models. The wear rates for initial and optimal models were recorded as 23.2 mm3/million cycles and 16.7 mm3/million cycles, respectively. Moreover, the experimental wear rate in the optimal design was 17.8 mm3/million cycles, which validated our optimization procedure. This study suggests that tibial insert conformity is an important factor in influencing the wear performance of patient-specific TKA, and it is capable of providing improved clinical results through enhanced design selections. This finding can boost the future development of patient-specific TKA, and it can be extended to other joint-replacement designs. However, further research is required to explore the potential clinical benefits of the improved wear performance demonstrated in this study.

Laser wavelength modulated pulsed laser ablation for selective and efficient production of graphene quantum dots.

Graphene quantum dots (GQDs) and graphene oxide quantum dots (GOQDs) can be used in different applications such as optoelectronic and biomedical applications, respectively. Hence, the selective synthesis of GQDs and GOQDs is highly desirable but challenging. Here, we present GQDs and GOQDs selectively prepared by an easy and simple pulsed laser ablation in liquid (PLAL) method by controlling the laser wavelength. The obtained GQDs and GOQDs showed a significantly different optoelectronic nature mainly due to the existence of surface oxygen-rich functional groups (e.g. carboxyl or hydroxy groups). Also, we described a possible mechanism for the formation of oxygen functional groups during the PLAL process based on the Coulomb explosion model, which can give further insight for designing functional carbon materials.

Simple preparation of graphene quantum dots with controllable surface states from graphite.

Graphite is economic and earth-abundant carbon precursor for preparing graphene quantum dots (GQDs). Here, we report a facile and green approach to produce GQDs from graphite flakes via a pulsed laser ablation (PLA) method assisted by high-power sonication. A homogeneous dispersion of graphite flakes, caused by high-power sonication during PLA, leads to the formation of GQDs following a laser fragmentation in liquid (LFL) rather than laser ablation in liquid (LAL) mechanism. The final product of GQDs exhibits the distinct structural, chemical, and optical properties of pristine graphene itself. However, graphene oxide quantum dots (GOQDs) with abundant surface oxygen-rich functional groups are readily formed from graphite flakes when high-power sonication is not employed during the PLA process. GQDs and GOQDs show a significantly different luminescence nature. Hence, selective production of either functional GQDs or GOQDs can be achieved by simply turning the high-power sonication during the PLA process on and off. We believe that our modified PLA process proposed in this work will further open up facile and simple routes for designing functional carbon materials.

Cadmium specifically induces MKP-1 expression via the glutathione depletion-mediated p38 MAPK activation in C6 glioma cells.

Cadmium is a toxic heavy metal and an environmental pollutant. Mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) is a negative regulator of the family of MAPK. In this study, we investigated the effect of heavy metals on MKP-1 expression in C6 rat glioma cells. Cadmium treatment induced MKP-1 at both protein and mRNA levels while cobalt or manganese treatment did not, suggesting the specificity. Cadmium treatment also depleted intracellular GSH and activated p38 MAPK, JNKs, and AKT. Profoundly, pretreatment with thiol-containing compounds NAC or GSH, but not vitamin E, blocked GSH depletion, 38 MAPK activation and MKP-1 expression by cadmium. Moreover, pharmacological inhibition of p38 MAPK by SB203580 suppressed the cadmium-induced MKP-1. Collectively, these results demonstrate that cadmium specifically induces MKP-1 by transcriptional up-regulation in C6 cells in a mechanism associated with the glutathione depletion-dependent p38 MAPK activation.

The effects of a new human leukocyte elastase inhibitor (recombinant guamerin) on cerulein-induced pancreatitis in rats.

BACKGROUND: Pancreatic and neutrophil elastase can aggravate or induce acute pancreatitis. Although increased elastase levels in the plasma of pancreatitis patients and animal models have been reported, the mechanism by which elastase is involved in the pathogenesis of acute pancreatitis has not yet been elucidated. We aimed to investigate the effects and the possible mechanism of a new human leukocyte elastase inhibitor (recombinant guamerin) in the treatment of cerulein-induced acute pancreatitis in rats. METHODS: Fifty Sprague-Dawley rats were divided into three groups: a saline-infused control group (I), a cerulein-induced acute pancreatitis group (II), and a cerulein plus guamerin infusion group (III). Guamerin (1-2 micromol/kg/h) was infused continuously in group III. The severity of pancreatitis was determined biochemically, histologically, and by cytokine changes between groups I, II and III. RESULTS: Significant differences in serum amylase, lipase, and pancreatic wet weight were observed in each group, respectively (group I; 2346.2 IU/L, 9.9 IU/L, 1.4+/-0.3 g, group II; 13,596.8 IU/L, 7439.4 IU/L, 2.2+/-0.5 g, group III; 9443.2 IU/L, 4516.3 IU/L, 1.7+/-0.6 g). Serum IL-6 and TNF-alpha [AU1]level peaked 1-4 h and 1-2 h. After the induction of pancreatitis, IL-6 and TNF-alpha levels were decreased in group III than group II, (group I; 13.1/4.0 pg/mL, group II; 198.5/63.2 pg/mL, group III; 102.1/13.1 pg/mL), but no significant difference in IL-1beta was observed. Histologic gradings and severity, such as vacuolization, inflammation, lobular disarray, and edema of the pancreas, were significantly lower in the cerulein plus guamerin infusion group III. CONCLUSIONS: Recombinant guamerin, a new human leukocyte elastase inhibitor, may decrease the severity of pancreatitis and diminish pancreatic acinar cell injury by inhibition of neutrophilic infiltration and cytokine activation in the initial stage of cerulein-induced acute pancreatitis in rats.

Activation of matrix metalloproteinase-9 by TNF-alpha in human urinary bladder cancer HT1376 cells: the role of MAP kinase signaling pathways.

The expression of matrix metalloproteinase-9 (MMP-9) has been implicated in tumor invasion and metastasis. In this study, the factors and signaling pathways that are involved in the regulation of the MMP-9 expression were examined in urinary bladder cancer HT1376 cells. Tumor necrosis factor-alpha (TNF-alpha) stimulated the secretion of MMP-9 in HT1376 cells, as shown by zymography and immunoblot analysis. At the level of transcription, TNF-alpha also stimulated 5'-flanking promoter activity of MMP-9. Transcription factor NF-kappaB, AP-1 and Sp-1 binding sites were identified by a gel shift assay to be cis-elements for TNF-alpha activation of the MMP-9 promoter. TNF-alpha activates multiple signaling pathways in HT1376 cells, including the extracellular signal-regulated kinase (ERK1/2), p38 MAP kinase and JNK pathways. Chemical inhibitors, which specifically inhibit each of these TNF-alpha-activated pathways, were used to examine the signaling pathways involved in TNF-alpha-mediated MMP-9 expression. The ERK1/2 inhibitor, U0126 and the p38 MAP kinase inhibitor, SB203580, significantly down-regulated TNF-alpha-induced MMP-9 expression and promoter activity. The transactivation of TNF-alpha-stimulated NF-kappaB, AP-1 and Sp-1 were inhibited by U0126 and SB203580 treatment. In conclusion, the findings of the present study indicate that TNF-alpha induces MMP-9 expression in HT1376 cells by activating transcription factors, which are involved in the ERK1/2- and p38 MAP kinase-mediated control of MMP-9 regulation, namely, NF-kappaB, AP-1 and Sp-1.

Fluoroscopically guided peritoneal dialysis catheter placement: long-term results from a single center.

OBJECTIVE: Despite percutaneous fluoroscopy ensuring appropriate placement of peritoneal dialysis (PD) catheters, the efficacy of this method is not well known. Therefore, we evaluated our long-term experience with fluoroscopy-assisted placement of PD catheters. PATIENTS AND METHODS: We retrospectively reviewed 134 PD catheters in 114 PD patients that were treated in the PD center of a university-based hospital. We evaluated complications related to PD catheters, causes for catheter removal, and catheter survival. We used the multivariate Cox proportional hazard model to identify independent factors related to PD catheter survival. RESULTS: Early complications related to insertion included 1 case of pericatheter bleeding; there were no placement failures. Early complications occurred in 8.5% of patients. Most late complications were migration and leakage, which occurred in 10.4% and 9.7% of patients respectively. The most common cause for catheter removal was intractable and recurrent peritonitis. The 12- and 24-month survival rates of the catheters were 80.0% and 74.9%. The most significant prognostic factor of percutaneous fluoroscopy-assisted PD catheter survival was late leakage (p < 0.01). CONCLUSIONS: In addition to the advantages of simplicity, minimal invasiveness, and relative safety, the survival rate of PD catheters placed using the percutaneous fluoroscopy-assisted method was comparable to that of more invasive methods. Percutaneous fluoroscopy-assisted placement of PD catheters should be considered when available, and may be preferred to other placement methods.