Confocal Laser Research Articles

A Novel Coated Suture Displays Antimicrobial Activity Without Compromising Structural Properties

BackgroundTreated or coated sutures promise to prevent contamination of wounds. PurposePurpose of the study was to coat surgical sutures with a new quaternary ammonium silane (QAS) antimicrobial compound at two different application temperatures and then to evaluate resulting structural, physical, mechanical, and biological properties. Study Design, Setting and SampleIn-vitro and in-vivo studies were conducted using male albino Wistar rats approved by Joint Ethical Committee of IMU and Postgraduate Medical Institute, Lahore. Only suture samples, coated uniformly with verified presence of the compound and of adequate length were used. Samples which were not coated uniformly and with inadequate length or damaged were excluded. Predictor VariablesPredictor variables were sutures with and without QAS coatings and different temperatures. Sutures were coated with QAS at 0.5% and 1.0% wt/vol using dip coating technique and sutures with and without QAS coating were tested at 25oC and 40oC temperatures. Main Outcome VariableOutcome variables of structural and physico-mechanical, properties of QAS coated and non-coated sutures were measured using Fourier transform infrared spectroscopy (FTIR) (for structural changes), confocal laser and scanning electron (for diameter changes), and tensile strength/modulus (for mechanical testing). Biologic outcome variables were tested (bacterial viability); macrophage cultures from Wistar rats were tested (M1/M2 polarization detecting IL-6 and IL-10). Macrophage cells were analyzed with CD80+ (M1) and CD163+ (M2). Chemotaxis Index was calculated as a ratio of quantitative fluorescence of cells. CovariatesNot applicable. AnalysisOrdinal data among groups were compared using Wilcoxon Mann-Whitney U test along with comparison of histological analysis using Wilcoxon Sign-rank test (p < 0.05). ResultsFTIR peak at 1490 cm-1 confirmed presence of QAS on suture’s surfaces with significant increase (p<0.05) in diameter (0.99±0.5-mm) and weight (0.77±0.02-mg) observed for 1% QAS-groups treated at 40°C. Non-coated samples heated at 25°C had significantly (p<0.05) less diameters (0.22±0.03-mm) and weights (0.26±0.06-mg). Highest tensile strength/modulus was observed for 0.5% QAS-coated samples which also had significantly high antibacterial characteristics than other sutures (p<0.05). QAS-coated sutures significantly increased M1 and M2 markers. Conclusions and RelevanceQAS coating conferred antibacterial action properties without compromising physical and mechanical properties of the suture.

M1 macrophage-derived exosome for reprograming M2 macrophages and combining endogenous NO gas therapy with enhanced photodynamic synergistic therapy in colorectal cancer

Reprogramming immunosuppressive M2 macrophages into M1 macrophages in tumor site provides a new strategy for the immunotherapy of colorectal cancer. In this study, M1 macrophage-derived exosome nanoprobe (M1UC) with Ce6-loaded upconversion material is designed to enhance the photodynamic performance of Ce6 while reprogramming M2 macrophages at tumor site and producing NO gas for three-mode synergistic therapy. Under the excitation of near-infrared light at 808 nm, the probe can generate 660 nm up-conversion fluorescence, which enables the photosensitizer Ce6 to produce ROS efficiently. In addition, the probe leads the production of NO by nitric oxide synthase on exosomes. Confocal laser and flow cytometry results show that M1UC probe reprograms M2 macrophages into M1 macrophages with an efficiency of 95.12%. The cell experiments show that the apoptosis rate of the three-mode synergistic therapy group is 78.8%, and the therapeutic effect is significantly higher than those of the other single treatment groups. In vivo experiments results show that M1UC probes maximally gather at the tumor site after 12 h of intravenous injection in orthotopic colorectal cancer mice. After 808 nm laser irradiation, the survival rate of mice is 100% and the recurrence rate was 0 within 60 d, and the therapeutic effect is significantly higher than those of other single treatment groups, which is also confirmed by immunohistochemistry. This M1 macrophage-derived exosome nanoplatform which is based on the three modes of immunotherapy, gas therapy and photodynamic therapy, provides a new design idea for the diagnosis and treatment of deep tumors.

Hypoxic Extracellular Matrix Preserves Its Competence after Expansion of Human MSCs under Physiological Hypoxia In Vitro.

Tissue-relevant O2 levels are considered as an important tool for the preconditioning of multipotent mesenchymal stromal cells (MSCs) for regenerative medicine needs. The present study investigated the quality and functions of the extracellular matrix (ECM) of MSCs under low O2 levels. Human adipose tissue-derived MSCs were continuously expanded under normoxia (20% O2, N) or "physiological" hypoxia (5% O2, Hyp). Decellularized ECM (dcECM) was prepared. The structure of the dcECM was analyzed using confocal laser and scanning electron microscopy. Collagen, dcECM-N, and dcECM-Hyp were recellularized with MSC-N and further cultured at normoxia. The efficacy of adhesion, spreading, growth, osteogenic potential, and paracrine activity of recellularized MSC-N were evaluated. At low O2, the dcECM showed an increased alignment of fibrillar structures and provided accelerated spreading of MSC-N, indicating increased dcECM-Hyp stiffness. We described O2-dependent "ECM-education" of MSC-N when cultured on dcECM-Hyp. This was manifested as attenuated spontaneous osteo-commitment, increased susceptibility to osteo-induction, and a shift in the paracrine profile. It has been suggested that the ECM after physiological hypoxia is able to ensure the maintenance of a low-commitment state of MSCs. DcECM, which preserves the competence of the natural microenvironment of cells and is capable of "educating" others, appears to be a prospective tool for guiding cell modifications for cell therapy and tissue engineering.

MP60-03 ORGANO-SELENIUM URINARY CATHETERS INHIBIT GROWTH AND BIOFILM FORMATION OF E. COLI , K. PNEUMONIAE , P. AERUGINOSA, AND H. INFLUENZAE IN-VITRO

You have accessJournal of UrologyCME1 Apr 2023MP60-03 ORGANO-SELENIUM URINARY CATHETERS INHIBIT GROWTH AND BIOFILM FORMATION OF E. COLI, K. PNEUMONIAE, P. AERUGINOSA, AND H. INFLUENZAE IN-VITRO Rachel Vopni, Unique Jacobo, Phat Tran, Shruti Patel, Suyash Jain, Cornelia De Riese, Ted Reid, and Werner De Riese Rachel VopniRachel Vopni More articles by this author , Unique JacoboUnique Jacobo More articles by this author , Phat TranPhat Tran More articles by this author , Shruti PatelShruti Patel More articles by this author , Suyash JainSuyash Jain More articles by this author , Cornelia De RieseCornelia De Riese More articles by this author , Ted ReidTed Reid More articles by this author , and Werner De RieseWerner De Riese More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003318.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Catheter-associated urinary tract infections (CAUTIs) are of significant medical burden in cost, morbidity, and mortality. Experimental selenium-coated medical devices have demonstrated nontoxic in-vitro and in-vivo antimicrobial activity due to the ability of selenium to catalyze formation of superoxide radicals. While antimicrobial-coated catheters have shown efficacy in preventing CAUTIs, selenium has not been tested in this context. The purpose of this in-vitro study is to evaluate selenium-incorporated urinary catheters for inhibition of uropathogenic bacterial growth and biofilm formation. METHODS: Urinary catheters incorporated with 1% organo-selenium and standard (uncoated) catheters were incubated in-vitro with E. coli, K. pneumoniae, P. aeruginosa, H. influenzae, and combinations of these bacteria. Growth was evaluated by colony forming unit count and visualized with confocal laser and scanning electron microscopy. Organo-selenium catheter material integrity was also tested by soaking the tubing in phosphate-buffered saline for 12-weeks at 37°C. RESULTS: Organo-selenium-incorporated catheters demonstrated total reduction (100%) of in-vitro bacterial growth and biofilm formation for E. coli, K. pneumoniae, H. influenzae, and a combination of these species when compared to control. P. aeruginosa growth was inhibited by approximately 4 logs (99.99%). Complete inhibition of E. coli growth was maintained after long-term phosphate-buffered saline soaking. CONCLUSIONS: The results demonstrate that organo-selenium was stably incorporated into catheter tubing and inhibited bacterial attachment, growth, and biofilm formation for multiple uropathogenic organisms. Furthermore, long-term soaking of organo-selenium tubing in phosphate-buffered saline did not show any decline in bacterial growth inhibition or biofilm formation. These findings suggest that organo-selenium-incorporated catheters may be advantageous in preventing CAUTIs and warrant further in-vivo and clinical evaluation. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e843 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rachel Vopni More articles by this author Unique Jacobo More articles by this author Phat Tran More articles by this author Shruti Patel More articles by this author Suyash Jain More articles by this author Cornelia De Riese More articles by this author Ted Reid More articles by this author Werner De Riese More articles by this author Expand All Advertisement PDF downloadLoading ...

Efficacy of organo-selenium-incorporated urinary catheter tubing for in vitro growth inhibition of E. coli, K. pneumoniae, P. aeruginosa, and H. influenzae.

Catheter-associated urinary tract infections are of significant medical burden in cost, morbidity, and mortality. Experimental selenium-coated medical devices have demonstrated non-toxic in vitro and in vivo antimicrobial activity. While antimicrobial-coated catheters have shown efficacy in preventing CAUTIs, selenium has not been tested in this context. The purpose of this in vitro study is to evaluate selenium-incorporated urinary catheters for inhibition of uropathogenic bacterial growth and biofilm formation. Urinary catheters incorporated with 1% organo-selenium and standard (uncoated) catheters were incubated in vitro with E. coli, K. pneumoniae, P. aeruginosa, H. influenzae, and combinations of these bacteria. Growth was evaluated by colony-forming unit count and visualized with confocal laser and scanning electron microscopy. Organo-selenium catheter material integrity was also tested by soaking the tubing in phosphate-buffered saline for 12weeks at 37°C. Organo-selenium-incorporated catheters demonstrated total reduction (100%) of in vitro bacterial growth and biofilm formation for E. coli, K. pneumoniae, H. influenzae, and a combination of these species when compared to control. P. aeruginosa growth was inhibited by approximately 4 logs (99.99%). Complete inhibition of E. coli growth was maintained after long-term phosphate-buffered saline soaking. The results demonstrate that organo-selenium was stably incorporated into catheter tubing and inhibited bacterial attachment, growth, and biofilm formation for multiple uropathogenic organisms. Furthermore, long-term soaking of organo-selenium tubing in phosphate-buffered saline did not show any decline in bacterial growth inhibition or biofilm formation. These findings suggest that organo-selenium-incorporated catheters may be advantageous in preventing catheter-associated urinary tract infections and warrant further in vivo and clinical evaluation.

Ginsenosides Rg1 and CK Control Temozolomide Resistance in Glioblastoma Cells by Modulating Cholesterol Efflux and Lipid Raft Distribution.

Background Cholesterol efflux and lipid raft redistribution contribute to attenuating temozolomide resistance of glioblastoma. Ginsenosides are demonstrated to modify cholesterol metabolism and lipid raft distribution, and the brain distribution and central nervous effects of whose isoforms Rb1, Rg1, Rg3, and CK have been identified. This study aimed to reveal the role of Rb1, Rg1, Rg3, and CK in the drug resistance of glioblastoma. Methods The effects of ginsenosides on cholesterol metabolism in temozolomide-resistant U251 glioblastoma cells were evaluated by cholesterol content and efflux assay, confocal laser, qRT-PCR, and Western blot. The roles of cholesterol and ginsenosides in temozolomide resistance were studied by CCK-8, flow cytometry, and Western blot, and the mechanism of ginsenosides attenuating resistance was confirmed by inhibitors. Results Cholesterol protected the survival of resistant U251 cells from temozolomide stress and upregulated multidrug resistance protein (MDR)1, which localizes in lipid rafts. Resistant cells tended to store cholesterol intracellularly, with limited cholesterol efflux and LXRα expression to maintain the distribution of lipid rafts. Ginsenosides Rb1, Rg1, Rg3, and CK reduced intracellular cholesterol and promoted cholesterol efflux in resistant cells, causing lipid rafts to accumulate in specific regions of the membrane. Rg1 and CK also upregulated LXRα expression and increased the cytotoxicity of temozolomide in the presence of cholesterol. We further found that cholesterol efflux induction, lipid raft redistribution, and temozolomide sensitization by Rg1 and CK were induced by stimulating LXRα. Conclusions Ginsenosides Rg1 and CK controlled temozolomide resistance in glioblastoma cells by regulating cholesterol metabolism, which are potential synergists for temozolomide therapy.

Atypical chorioretinal lesions in Siberian Husky dogs with primary angle-closure glaucoma: a case series

BackgroundA number of etiologies for different canine chorioretinal lesions have been proved or suggested but some fundic lesions remain unclear in terms of an etiologic diagnosis, treatment options and prognosis. The purpose of this case series is to describe atypical chorioretinal lesions observed in dogs with primary angle-closure glaucoma (PACG).Case presentationTwo spayed-female Siberian Huskies (3- and 4-year-old) and one Siberian Husky/Australian Shepherd mixed breed dog (11-month-old) that had multifocal depigmented retinal lesions and PACG were included. Procedures: Ophthalmic examination, gross, and histopathologic examination findings are described. One of the dogs underwent further clinical diagnostics. Advanced clinical diagnostics on the fellow, presumed to be non-glaucomatous eye of a dog revealed: pectinate ligament dysplasia by gonioscopy, retinal thinning in the depigmented area and wedge shaped retinal thinning with delayed choroidal vascular perfusion by optical coherence tomography, confocal scanning laser ophthalmoscopy, fluorescein and indocyanine green angiography. Quantifiable maze testing for the same eye revealed mild nyctalopia but the full-field electroretinogram showed no generalized decrease of retinal function. Genetic testing for mutations within the retinitis pigmentosa GTPase regulator gene causing X-linked progressive retinal atrophy in Siberian Huskies was negative. Histopathologic evaluations on enucleated eyes in two dogs confirmed goniodysgenesis, PACG with optic nerve head cupping, and diffuse inner retinal atrophy. In addition, segmental profound retinal atrophy, loss of retinal pigment epithelium, and adhesion of the retina to Bruch’s membrane was observed and coincided with multifocal depigmented lesions noted on fundic examination.ConclusionsTo our knowledge, this is the first case series with clinical and histopathologic data of chorioretinal lesions, most likely caused by severely impaired choroidal perfusion. Further studies are warranted to elucidate the etiology and pathophysiology, including its possible association with PACG.

Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol.

Several types of deproteinised bovine bone mineral (DBBM) are available on the market, and each one is obtained with a thermic and chemical process that can differ, achieving different results. Currently, several protocols using low temperature are suggested to reduce the possible particle crystallisation during the production process. This study aimed to evaluate the biomorphological reaction of periodontal fibroblast cultures in contact with different DBBM particles treated with a low-temperature protocol (Thermagen®) and without exposure to sodium hydroxide (NaOH). Morphological evaluation was performed using light, confocal laser, and scanning electron microscopy, and the biological reaction in terms of proliferation was performed using an XTT proliferation assay at 24 h (T1), 72 h (T2), and 7 days (T3). The morphological analysis highlighted how the presence of the materials stimulated a change in the morphology of the cells into a polygonal shape, surface reactions with the thickening of the membrane, and expression of actin. In particular, the morphological changes were appreciable from T1, with a progressive increase in the considered morphological characteristics at T2 and T3 follow-ups. The proliferation assay showed a statistical significance between the different experimental materials and the negative control in T2 and T3 follow-ups. The post hoc analysis did not reveal any differences between the materials. In conclusion, the grafts obtained with the low-temperature extractions protocol and not exposed to NaOH solution showed positive morphological reactions with no differences in the sizes of particles.

Asymmetry of Optic Nerve Head Parameters Measured by Confocal Scanning Laser Ophthalmoscopy in Myopic Anisometropic Eyes

Background: This study aimed to evaluate the asymmetry of optic nerve head parameters measured by confocal scanning laser ophthalmoscopy (CSLO) in myopic anisometropic eyes. Methods: A total of 36 eyes of 18 healthy myopic anisometropic subjects, defined as cases in which the difference in spherical equivalent (SE) between both eyes is ≥1.5D, were recruited. The optic nerve heads were measured using the Heidelberg retina tomograph II (Heidelberg Engineering, GmBH, Heidelberg, Germany). Differences in optic nerve head parameters between more myopic eyes and fellow eyes were evaluated using the paired-sample t-test. Pearson correlation and multiple linear regression analysis were used to evaluate factors associated with cup/disc ratio (CDR). Results: The cup/disc area ratio (mean difference 0.07 ± 0.11, P = 0.027), horizontal (mean difference 0.10 ± 0.17, P = 0.033), and vertical CDR (mean difference 0.13 ± 0.18, P = 0.008) were significantly smaller in more myopic eye. Larger disc area was independently and significantly associated with larger cup/disc area ratio (β = 0.561, P = 0.001) and vertical CDR (β = 0.499, P = 0.03). Conclusion: The CDR, horizontal, and vertical CDR were significantly smaller in the more myopic eyes in myopic anisometropic subjects. Further studies with larger samples are needed to confirm the asymmetry of the optic nerve head in myopic anisometropic eyes.

A hyper-resolving polynomial aperture and its application in microscopy

BackgroundA hyper-resolving aperture composed of a polynomial distribution is suggested. The point spread function (PSF) is computed and compared with that corresponding to linear, quadratic, and circular apertures. In addition, the influence of the number of zones on the PSF is discussed. An application on confocal scanning laser microscope using Siemen’s star pattern as an object considering the polynomial apertures is given.ResultsWe have made polynomial apertures using MATLAB code, and we tested the resolution from the computation of the cut-off spatial frequency obtained from the computation of the point spread function.ConclusionsWe get compromised resolution and contrast for the polynomial apertures as compared with uniform circular apertures.

Globular adiponectin protects hepatocytes against intermittent hypoxia-induced injury via Pink1/Parkin-mediated mitophagy induction.

This study sought to determine the effect of Pink1/Parkin-mediated mitophagy on liver cells exposed to intermittent hypoxia (IH) and the roles of globular adiponectin (gAPN). The hepatocyte model of IH was established. Cell apoptosis was assessed using flow cytometry. Mitochondrial membrane potential (MMP) level was determined using JC-1, and mitophagy was assessed using a confocal laser. Mitochondrial injury associated protein levels of bax and bcl-2, and protein levels of Pink1 and Parkin were evaluated via western blotting. We downregulated Parkin expression by transfecting the cells with Parkin siRNA. Pink1 and Parkin protein levels, mitophagy, and cell apoptosis rate were high, while the MMP level and protein level ratio of bcl-2/bax were low in IH-treated hepatocyte. gAPN upregulated Pink1 and Parkin protein levels, MMP level, protein level ratio of bcl-2/bax, and mitophagy while it reduced the rate of cell apoptosis in IH-treated hepatocytes. Inhibiting Parkin expression significantly reduced mitophagy and increased mitochondrial injury andthe rate of hepatocyte apoptosis under IH or IH with gAPN. gAPN alleviated IH-induced mitochondrial injury and hepatocyte apoptosis by upregulating Pink1/Parkin-mediated mitophagy.

Optical coherence tomography-measured retinal nerve fiber layer thickness values compensated with a multivariate model and discrimination between stable and progressing glaucoma suspects

PurposeOur previously introduced multivariate model, compensating for intersubject variability, was applied to circumpapillary retinal nerve fiber layer (RNFL) values measured with optical coherence tomography in glaucoma suspects with or without prior progressive optic disc (OD) change in a series of confocal scanning laser tomography (CSLT, HRT III) measurements.MethodsIn this prospective study, OD change during CSLT follow-up was determined with strict, moderate, and liberal criteria of the topographic change analysis (TCA). Model compensation (MC) as well as age compensation (AC) was applied to RNFL sectors (RNFLMC vs. RNFLAC). Diagnostic performance of RNFLMC vs. RNFLAC was tested with an area under the receiver operating characteristic (AUROC) and was compared between methods.ResultsForty-two glaucoma suspects were included. Patients without prior progressive OD change during the CSLT follow-up (= stable) had thicker RNFL thickness values in most areas and for all progression criteria. RNFLMC AUROC for the global RNFL (0.719) and the inferior quadrant (0.711) performed significantly better compared with RNFLAC AUROC (0.594 and 0.631) to discriminate between stable and progressive glaucoma suspects as defined by the moderate criteria of CSLT progression analysis (p = 0.028; p = 0.024).ConclusionMC showed a slight but significant improvement in detection of subjects with prior progressive OD change among a group of glaucoma suspects, when compared to AC, which is the compensation method commonly used during OCT data evaluation in daily routine. Further studies are warranted to validate the present results.

Confocal spectral microscopy, a non-destructive approach to follow contamination and biofilm formation of mCherry Staphylococcus aureus on solid surfaces

Methods to test the safety of wood material for hygienically sensitive places are indirect, destructive and limited to incomplete microbial recovery via swabbing, brushing and elution-based techniques. Therefore, we chose mCherry Staphylococcus aureus as a model bacterium for solid and porous surface contamination. Confocal spectral laser microscope (CSLM) was employed to characterize and use the autofluorescence of Sessile oak (Quercus petraea), Douglas fir (Pseudotsuga menziesii) and poplar (Populus euramericana alba L.) wood discs cut into transversal (RT) and tangential (LT) planes. The red fluorescent area occupied by bacteria was differentiated from that of wood, which represented the bacterial quantification, survival and bio-distribution on surfaces from one hour to one week after inoculation. More bacteria were present near the surface on LT face wood as compared to RT and they persisted throughout the study period. Furthermore, this innovative methodology identified that S. aureus formed a dense biofilm on melamine but not on oak wood in similar inoculation and growth conditions. Conclusively, the endogenous fluorescence of materials and the model bacterium permitted direct quantification of surface contamination by using CSLM and it is a promising tool for hygienic safety evaluation.

Host–pathogen interaction between macrophage co-cultures with Staphylococcus aureus biofilms

The ability of Staphylococcus aureus to form biofilms is an important virulence factor. During the infectious process, the interaction between biofilms and immune cells is determinant; however, the properties that make biofilms resistant to the immune system are not well characterized. In order to better understand this, we evaluated the in vitro interaction of macrophages during the early stages of S. aureus biofilm formation. Biofilm formation was evaluated by crystal violet staining, light microscopy, and confocal scanning laser microscopy. Furthermore, different activation on L-arginine pathways such as nitric oxide (NO•) release and the arginase, the production of reactive oxygen species (ROS), the total oxidative stress response (OSR), and levels of cytokine liberation, were determined. Our findings show that the interaction between biofilms and macrophages results in stimuli for catabolism of L-arginine via arginase, but not for NO•, an increase of ROS production, and activation of the non-enzymatic OSR. We also observed the production of IL-6, but not of TNFα o IL-10 in these co-cultures. These results contribute to a better understanding of host-pathogen interactions and suggest that biofilms increase resistance against immune cell mechanisms, a phenomenon that could contribute to the ability of S. aureus biofilms to establish mature biofilms.

Iso- and Anisotropic Etching of Micro Nanofibrillated Cellulose Films by Sequential Oxygen and Nitrogen Gas Plasma Exposure for Tunable Wettability on Crystalline and Amorphous Regions.

The surface of cellulose films, obtained from micro nanofibrillated cellulose produced with different enzymatic pretreatment digestion times of refined pulp, was exposed to gas plasma, resulting in a range of surface chemical and morphological changes affecting the mechanical and surface interactional properties. The action of separate and dual exposure to oxygen and nitrogen cold dielectric barrier discharge plasma was studied with respect to the generation of roughness (confocal laser and atomic force microscopy), nanostructural and chemical changes on the cellulose film surface, and their combined effect on wettability. Elemental analysis showed that with longer enzymatic pretreatment time the wetting response was sensitive to the chemical and morphological changes induced by both plasma gases, but distinctly oxygen plasma was seen to induce much greater morphological change while nitrogen plasma contributed more to chemical modification of the film surface. In this novel study, it is shown that exposure to oxygen plasma, subsequently followed by exposure to nitrogen plasma, leads first to an increase in wetting, and second to more hydrophobic behaviour, thus improving, for example, suitability for printing using polar functional inks or providing film barrier properties, respectively.

Corneal hysteresis as a risk factor for optic nerve head surface depression and retinal nerve fiber layer thinning in glaucoma patients

To evaluate the role of corneal hysteresis (CH) as a risk factor for progressive ONH surface depression and RNFL thinning measured by confocal scanning laser ophthalmoscopy (CSLO) and spectral-domain optical coherence tomography (SD-OCT), respectively in glaucoma patients. Prospective study. A total of 146 eyes of 90 patients with glaucoma were recruited consecutively. The CH measurements were acquired at baseline and 4-months interval using the Ocular Response Analyzer (Reichert Instruments, Depew, NY). Eyes were imaged by CSLO (Heidelberg Retinal Tomograph [HRT]; Heidelberg Engineering, GmbH, Dossenheim, Germany) and SD-OCT (Cirrus HD-OCT; Carl Zeiss Meditec AG, Dublin, CA) at approximately 4-month intervals for measurement of ONH surface topography and RNFL thickness, respectively. Significant ONH surface depression and RNFL thinning were defined with reference to Topographic Change Analysis (TCA) with HRT and Guided Progression Analysis (GPA) with Cirrus HD-OCT, respectively. Multivariate cox proportional hazards models were used to investigate whether CH is a risk factor for ONH surface depression and RNFL progression after adjusting potential confounding factors. All patients with glaucoma were followed for an average of 6.76 years (range, 4.56–7.61 years). Sixty-five glaucomatous eyes (44.5%) of 49 patients showed ONH surface depression, 55 eyes (37.7%) of 43 patients had progressive RNFL thinning and 20 eyes (13.7%) of 17 patients had visual field progression. In the cox proportional hazards model, after adjusting baseline diastolic IOP, CCT, age, baseline disc area and baseline MD, baseline CH was significantly associated with ONH surface depression and visual field progression (HR = 0.71, P = 0.014 and HR = 0.54, P = 0.018, respectively), but not with RNFL thinning (HR = 1.03, P = 0.836). For each 1-mmHg decrease in baseline CH, the hazards for ONH surface depression increase by 29%, and the hazards for visual field progression increase by 46%. The CH measurements were significantly associated with risk of glaucoma progression. Eyes with a lower CH were significantly associated with an increased risk of ONH surface depression and visual field progression in glaucoma patients.

Increasing the photocatalytic and fungicide activities of Ag3PO4 microcrystals under visible-light irradiation

The present study reports for the first time the performance of silver phosphate (Ag3PO4) microcrystals as photocatalyst (degradation of Rodamine B-RhB) and antifungal agent (against Candida albicans–C. albicans) under visible-light irradiation (455 nm). Ag3PO4 microcrystals were synthesized by a simple co-precipitation (CP) method at room temperature. The structural and electronic properties of the as-synthetized Ag3PO4 have been investigated before and after 4 cycles of RhB degradation under visible light using X-ray diffraction (XRD), micro-Raman spectroscopy, UV–Vis spectrophotometer and field emission scanning electron microscopy (FE-SEM) images. The antifungal activity was analyzed in planktonic cells and 48h-biofilm of C. albicans by colony forming units (CFU) counting, confocal laser and FE-SE microscopies. Statistical analysis was carried out using SPSS software. Morphological and structural modifications of Ag3PO4 were observed upon recycling. After 4 recycles, the material maintained its photodegradation property; an eightfold increase in the efficiency of Ag3PO4 was observed in planktonic cells and a two fold increase in biofilm when irradiated under visible light. Thus, higher antifungal effectiveness against C. albicans was obtained when associated with visible-light irradiation.

Isothermal and Non-isothermal Crystallization Kinetics of Mold Fluxes used in Continuous Casting of Steel: A Review

Casting powders or mold fluxes, as they are more commonly known, are used in the continuous casting of steel to prevent the steel shell from sticking to the copper mold. The powders first melt and create a pool of liquid flux above the liquid steel in the mold, and then the liquid mold fluxes penetrate into the gap between water-cooled copper mold and steel shell, where crystallization of solid phases takes place as the temperatures gradually drop. It is important to understand the crystallization behavior of these mold fluxes used in the continuous casting of steel because the crystalline phase fraction in the slag films plays a crucial role in determining the horizontal heat flux during the casting process. In this work, the existing literature on the crystallization kinetics of conventional and fluoride-free mold fluxes used in the continuous casting of steel has been reviewed. The review has been divided into two main sections viz. the isothermal crystallization kinetics and non-isothermal crystallization kinetics. Under each of these sections, three of the most widely used techniques for studying the crystallization kinetics have been included viz. thermoanalytical techniques such as differential scanning calorimetry/differential thermal analysis (DSC/DTA), the single and double hot thermocouple technique (SHTT and DHTT), and the confocal scanning laser microscopy (CSLM). For each of these techniques, the available literature related to the crystallization kinetics of mold fluxes has been summarized thereby encompassing a wide range of investigations comprising of both conventional and fluoride-free fluxes. Summaries have been included after each section with critical comments and insights by the authors. Finally, the relative merits and demerits of these methods vis-à-vis their application in studying the crystallization kinetics of mold fluxes have been discussed.

Could the Calcium Silicate-Based Sealer Presentation Form Influence Dentinal Sealing? An In Vitro Confocal Laser Study on Tubular Penetration.

Dentinal tubule penetration influences root canal treatment sealing. The aim of this study was to compare dentinal penetration of two clinical presentations of silicate-based sealers using confocal laser. Sixty single-rooted human teeth from 50–70 year-old patients extracted for orthodontic/periodontal reasons were used. Canals were prepared using Mtwo system up to 35/0.04, with 5.25% NaOCl irrigation and final irrigation using 17% EDTA. Teeth were randomly assigned into study groups (ni = 20): EndoSequence BC sealer (ES, group 1), BioRoot RCS (BR, group 2); and a control group (nc = 20) with AH Plus (AHP). Root canals were obturated with 35/0.04 gutta-percha (single-cone technique). The samples were obtained from apical, middle, and coronal thirds. Dentinal tubule penetration depth and percentage of penetration around the canal perimeter were measured. The statistical analysis was performed using Mann Whitney U test and Wilcoxon t-test (95% confidence interval). ES exhibited a significantly higher penetration than AHP in apical and middle thirds (p < 0.05), and in middle and coronal thirds relative to BR (p < 0.05). The percentage of penetration around the canal perimeter was significantly higher for ES compared to BR in all thirds, but only in the apical third for AHP (p < 0.05). The pre-mixed silicate-based sealer exhibited better penetration than the powder/liquid one.

Time-dependent effects on physicochemical and surface properties of PHBV fibers and films in relation to their interactions with fibroblasts

Biodegradability or materials physicochemical stability are the key biomaterials selection parameters for various medical and tissue engineering applications. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a natural copolymer known from its biocompatibility with great support for cells growth and attachment on films and fibers. In our studies, the physicochemical properties of electrospun PHBV fibers and spin-coated films aged for 1, 4 and 8 weeks were analyzed using bulk (FTIR) and surface chemistry (XPS) methods and water contact angle. Further, we characterized the zeta potential changes after aging, by means of electrokinetic measurements, and cell responses to it, using NIH 3T3 murine fibroblasts. Colorimetric MTS cell viability test allowed the assessment of cell proliferation. Additionally, the morphology of fibroblasts and biointerfaces were studied by confocal laser and electron scanning microscopy (CLSM and SEM). These studies indicated that the activity, attachment and proliferation of fibroblasts is independent of aging of PHBV fibers and films. PHBV films show very stable zeta potential over 8 weeks of aging, opposite to PHBV fibers. Importantly, the flat film of PHBV increases cell proliferation, while the fibrous meshes are an excellent support for their stretching. The results of the study revealed clear advantages of PHBV films and fibrous meshes in cell-material interaction.