Normal Lens Research Articles

In Vivo Observation of Lens Regeneration in Rat Using Ultra-Long Scan Depth Optical Coherence Tomography.

To evaluate morphologic changes of lens regeneration in rats in vivo after extracapsular lens extraction (ECLE) by ultra-long scan depth optical coherence tomography (UL-OCT). A total of 42 Sprague-Dawley rats were used in this study. We performed ECLE on the right eyes of animals in the surgery group (n = 34). Biomicroscopy and UL-OCT scans were carried out for the surgery group immediately (within 1 hour postoperatively) and at days 1 and 3, weeks 1 and 2, and months 1, 2, and 3 postoperatively. After in vivo examination, three animals of the surgery group were euthanized at each time point for histology study, while the other 10 animals were examined continuously at those time points. The regenerated lens was evaluated in OCT images at 2 and 3 months postoperatively. The control group consisted of eight untreated rats that had OCT examination at the age of 5 months. Lens regeneration could be observed from 2 weeks postoperatively. Regeneration was mainly at the peripheral capsular bag in the first month and central region thereafter. The average thickness of regenerated lenses was 2222 ± 309 and 2324 ± 352 μm at 2 and 3 months, respectively. Regeneration was faster in the first 2 months and slowed down thereafter. Although anterior capsule opening and posterior capsule adhesion and wrinkling existed, the regenerated lens still could form a relatively regular shape, however, the size was much smaller than that of the normal lenses from rats with the same age. Ultra-long OCT provides longitudinal data of the process of lens regeneration on a single individual rat in vivo, which may allow one to follow and compare the lens regenerative process under different interventions or therapy after ECLE in rats.

Spatial distributions of glutathione and its endogenous conjugates in normal bovine lens and a model of lens aging

Glutathione (GSH) is the archetypal antioxidant, and plays a central role in the protection of the ocular lens from cataract formation. High levels of GSH are maintained in the transparent lens, but with advancing age, GSH levels fall in the lens nucleus relative to outer cortical cells, thereby exposing the nucleus of the lens to the damaging effects of oxygen radicals, which ultimately leads to age-related nuclear (ARN) cataract. Under normal conditions, GSH also forms endogenous conjugates to detoxify the lens of reactive cellular metabolites and to maintain cell homeostasis. Due to the intrinsic gradient of lens fibre cell age, the lens contains distinct regions with different metabolic requirements for GSH. To investigate the impact of fibre cell and lens aging on the varied roles that GSH plays in the lens, we have utilised high mass resolution MALDI mass spectrometry profiling and imaging analysis of lens tissue sections. High Dynamic Range (HDR)-MALDI FTICR mass spectrometry was used as an initial screening method to detect regional differences in lens metabolites from normal bovine lenses and in those subjected to hyperbaric oxygen as a model of lens aging. Subsequent MALDI imaging analysis was used to spatially map GSH and its endogenous conjugates throughout all lenses. Accurate mass measurement by MALDI FTICR analysis and LC-MS/MS mass spectrometry of lens region homogenates were subsequently used to identify endogenous GSH conjugates. While the distribution and relative abundance of GSH-related metabolic intermediates involved in detoxification pathways remained relatively unchanged upon HBO treatment, those involved in its antioxidant function were altered under conditions of oxidative stress. For example, reduced glutathione levels were decreased in the lens cortex while oxidised glutathione levels were elevated in the lens outer cortex upon HBO treatment. Interestingly, cysteineglutathione disulfide, was detected in the inner cortex of the normal lens, but was greatly decreased in the HBO-treated lenses. These results contribute to our understanding of the multiple roles that GSH plays in maintenance of lens transparency and in the age-related metabolic changes that lead to lens cataract formation.

Hepatoprotective effect of Ginkgo biloba against intoxication caused by a single dose of carbon tetrachloride

To analyze the expression of 78 kDa glucose-regulated protein (GRP78) and activating transcription factor 6 (ATF6), 2 factors in the unfolded protein response (UPR), in age-related and diabetes-associated cataract.Universidad Autónoma de Aguascalientes, Aguascalientes, México.Experimental study.The qualitative and quantitative expression of GRP78 and ATF6 were measured in surgical samples from 11 senile cataracts, 9 diabetic-associated cataracts, and 3 normal lenses. Both proteins were detected by immunofluorescence and immunogold-conjugated antibodies. Quantitative morphometry was used to analyze the differences in GRP78 and ATF6 between samples. The Mann-Whitney test was used for statistical analysis.Scanning electron microscopy showed the characteristic organization of fibers in normal lenses with regular alignment and interdigitation between them. On the other hand, lenses from eyes with senile or diabetic cataract showed the same pattern of misalignment and disorganization of the fibers. Both proteins were detected through immunofluorescence in senile and diabetic cataracts, but not in normal lenses. Immunogold-conjugated antibodies and transmission electron microscopy showed that GRP78 and ATF6 grains were 30% higher and 35% higher, respectively, in diabetic cataracts than in senile cataracts (P<.05).These data show for the first time in humans that GRP78 and ATF6 are present in lens fibers of senile cataracts and diabetic cataracts, establishing that the UPR may be important in the process of cataractogenesis.No author has a financial or proprietary interest in any material or method mentioned.

Analysis on the Characteristics of Camera Lens Distortion

Objectives: Recent high zoom lens as CCD (Charge Coupled Device) camera is released with the resolution to be able to easily acquire the digital image that has been variously utilized, from day-to-day utilization to take advantage of specialized domain such as computer vision (computer vision). It has a number of advantages for obtaining a normal zoom lens CCD camera video which is now commercially available, however the camera calibration in the actual image process geometrically have been the considerable difficulties due to the unstable capture and the movement of the pick-up process in a variety of zoom lens camera. Methods: The camera parameters for the zoom lens calibration will be test variables calculated over a camera lens, a variety of focal lengths whenever the zoom occur, especially if you already have zoom movement at the point the lens test is complete, recalculation of the camera calibration parameters cause the difficulties. In this research the zoom lens to correct the distortion of image obtaining the camera calibration, and a method that extracts the camera parameters through the DLT (Direct Linear Transformation) test method is proposed. Findings: DLT accuracy of the camera is black was carried out in the following two aspects. The first was to analyze the three dimensional position differences between the estimated position and the actual three-dimensional observation that is calculated by the model formula to moderate, and the second is that pixel position that is pixels accuracy projected onto the image plane in the three-dimensional space of the object the accuracy was evaluated in an absolute manner. In this study, under the assumption that the aperture condition is fixed zoom, the zoom was determined the relationship between the camera and variable focus for the condition setting. A zoom lens camera model by setting the zoom and focus conditions at regular intervals as short-focus lens model was established by the DLT method and each camera parameter is tested individually. Applications: In future research the image correction software will be develop for the 3-dimensional location information based on three-dimensional information generating process based on camera calibration method by the DLT method.

CONVEX LENS WITH TRANSPARENT GLASS SOLAR WATER HEATING SYSTEM

Conventional energy solutions are necessary in developing countries as current hot water production methods are becoming hazardous to the environment, economic development and the overall health and wealth of the population. The reason of this research paper is to create a convex lens with transparent glass solar water heater prototype, experimentally analyze the system and predict its characteristics when system exposed to the solar behavior of Bangalore, Karnataka, India. Present design from manufacturer and supplier is plane transparent glass solar water heating system. A user of solar energy minimum because cost of the product. With the merits of small structure, less weight, sharp focal length and minimum cost convex lenses are suitable for concentration of solar radiation and system improvement. The efficiency and optical property of transmitted solar radiation is improved and controlled compared to thick normal glass lenses and the solar radiation tracking accuracy need for convex lens group is less compared to other concentration methods.

Comparison of several transport activities of lens epithelial cells from cataract and healthy dog

PurposeThe aim of this study is to compare the transport activities between normal and cataract lens epithelial cells.MethodsThe primary lens epithelial cells of healthy and cataract dog were transfected with the expression plasmid DNA of large T antigen from replication origin‐defective simian virus 40 (SV40), then cloned using a glass cylinder. The transport activities of glycine, cysteine, glutamate, arginine, leucine, carnitine, ascorbic acid and glutathione were investigated using two cell lines.ResultsThere was no significant difference in transport activity between the two cells lines except for glutamate. Glutamate transport activity was 20% lower in cells from cataract than healthy one.ConclusionsReduced glutamate transport activity may contribute to the pathogenesis of canine cataract.

Optical coherence tomography findings of retinal folds in nanophthalmos

SummaryNanophthalmos is a rare form of congenital microphthalmos and it can be seen in siblings due to genetic transmission. The eye is small in overall dimensions including short axial length (16–18.5 mm) with shallow anterior chamber and almost normal lens size. Typically high hyperopia is seen. Posterior findings include; cupless and crowded optic disc, and elevated papillomacular retinal folds. Optic Coherence Tomography (OCT) can reveal retinal fold with normal retinal pigment epithelium and choriocapillaris. It is presumed that the retinal folds in nanophthalmos result from a redundancy of the retinal layer caused by retarded growth of the scleral, choroidal, and retinal pigment epithelial layers and OCT can demonstrate papillomacular retinal folds that are confined to the neurosensory retina.

OPTIMIZATION OF LASER SURGERY OF THE SECONDARY CATARACT

Background: This is a theoretical and experimental study of the thermomechanical effects of laser radiation with a wavelength of 1.06 microns on the eye tissues during a laser surgery on the secondary cataract (pupillary membrane). Its relevance is related to the rates of complications after laser surgery of the eye associated with the choice of energy and time parameters of the laser irradiation. These parameters are related to the occurrence of such factors as unstable fields of thermal stress and pressure that are difficult to take into account and indirectly lead to adverse events when removing the secondary cataract. Aim: To minimize side effects and to optimize the existing technology of laser removal of the secondary cataract. Materials and methods: Samples of a normal lens capsule and of lens capsules with various types of opacities taken during a cataract surgery, with various optic characteristics and thickness, were treated with an infrared laser (Nd:YAG laser, 1064 nm). We performed morphometric measurements and built up a theoretical model of the processes in a continuous medium under the effects of impulse laser irradiation. Results: The results of numerical modelling with this newly developed theoretical model are in satisfactory agreement with the experimental data on development of deformities obtained with the autopsy materials (posterior capsule of the human lens with various optical characteristics and thickness, from thin transparent membranes to more thick opaque samples). Conclusion: This study would allow for optimization of the technology of laser treatment for secondary cataracts by changing the irradiation parameters during the procedure.

EVALUATION OF THE QUALITY OF ACTION CAMERAS WITH WIDE-ANGLE LENSES IN UAV PHOTOGRAMMETRY

Abstract. The application of light-weight cameras in UAV photogrammetry is required due to restrictions in payload. In general, consumer cameras with normal lens type are applied to a UAV system. The availability of action cameras, like the GoPro Hero4 Black, including a wide-angle lens (fish-eye lens) offers new perspectives in UAV projects. With these investigations, different calibration procedures for fish-eye lenses are evaluated in order to quantify their accuracy potential in UAV photogrammetry. Herewith the GoPro Hero4 is evaluated using different acquisition modes. It is investigated to which extent the standard calibration approaches in OpenCV or Agisoft PhotoScan/Lens can be applied to the evaluation processes in UAV photogrammetry. Therefore different calibration setups and processing procedures are assessed and discussed. Additionally a pre-correction of the initial distortion by GoPro Studio and its application to the photogrammetric purposes will be evaluated. An experimental setup with a set of control points and a prospective flight scenario is chosen to evaluate the processing results using Agisoft PhotoScan. Herewith it is analysed to which extent a pre-calibration and pre-correction of a GoPro Hero4 will reinforce the reliability and accuracy of a flight scenario.

EVALUATION OF THE QUALITY OF ACTION CAMERAS WITH WIDE-ANGLE LENSES IN UAV PHOTOGRAMMETRY

The application of light-weight cameras in UAV photogrammetry is required due to restrictions in payload. In general, consumer cameras with normal lens type are applied to a UAV system. The availability of action cameras, like the GoPro Hero4 Black, including a wide-angle lens (fish-eye lens) offers new perspectives in UAV projects. With these investigations, different calibration procedures for fish-eye lenses are evaluated in order to quantify their accuracy potential in UAV photogrammetry. Herewith the GoPro Hero4 is evaluated using different acquisition modes. It is investigated to which extent the standard calibration approaches in OpenCV or Agisoft PhotoScan/Lens can be applied to the evaluation processes in UAV photogrammetry. Therefore different calibration setups and processing procedures are assessed and discussed. Additionally a pre-correction of the initial distortion by GoPro Studio and its application to the photogrammetric purposes will be evaluated. An experimental setup with a set of control points and a prospective flight scenario is chosen to evaluate the processing results using Agisoft PhotoScan. Herewith it is analysed to which extent a pre-calibration and pre-correction of a GoPro Hero4 will reinforce the reliability and accuracy of a flight scenario.

MicroRNA let-7b induces lens epithelial cell apoptosis by targeting leucine-rich repeat containing G protein-coupled receptor 4 (Lgr4) in age-related cataract

Owing to a rapidly aging population, vision impairment caused by age-related cataract has become very common. Age-related cataract has also become one of the principal causes of blindness, and apoptosis of lens epithelial cells contributes to non-congenital cataract development. Previous studies have reported that microRNA let-7b (let-7b) is upregulated in cataractous lens epithelial cells, and the expression level of let-7b is positively associated with N, C and P cataract scores. However, the role of let-7b in the development of age-related cataract remains unclear. Here, we observed that the expression level of let-7b in the anterior lens capsules of age-related cataract was significantly higher than that in the normal anterior lens capsules. We performed ultraviolet (UV) irradiation to induce lens epithelial cell apoptosis. The results showed that the expression level of let-7b in lens epithelial cells which were treated by UV irradiation was significantly higher than that in the control, and let-7b promoted UV irradiation-induced apoptosis. Furthermore, we showed that leucine-rich repeat containing G protein-coupled receptor 4 (Lgr4) was a direct target of let-7b, and let-7b modulated lens epithelial cell apoptosis by directly targeting Lgr4. These findings will offer new insights into our understanding of the molecular mechanisms underlying the pathogenesis of cataract.

Intrinsic and extrinsic regulatory mechanisms are required to form and maintain a lens of the correct size and shape

Understanding how tissues and organs acquire and maintain an appropriate size and shape remains one of the most challenging areas in developmental biology. The eye lens represents an excellent system to provide insights into regulatory mechanisms because in addition to its relative simplicity in cellular composition (being made up of only two forms of cells, epithelial and fiber cells), these cells must become organized to generate the precise spheroidal arrangement that delivers normal lens function. Epithelial and fiber cells also represent spatially distinct proliferation and differentiation compartments, respectively, and an ongoing balance between these domains must be tightly regulated so that the lens achieves and maintains appropriate dimensions during growth and ageing. Recent research indicates that reciprocal inductive interactions mediated by Wnt-Frizzled and Notch-Jagged signaling pathways are important for maintaining and organizing these compartments. The Hippo-Yap pathway has also been implicated in maintaining the epithelial progenitor compartment and regulating growth processes. Thus, whilst some molecules and mechanisms have been identified, further work in this important area is needed to provide a clearer understanding of how lens size and shape is regulated.

Molecular Characterization of Two Human Lens Epithelial Cell Lines and Their Suitability to Study Function of Cataract Genes

The ocular lens is a transparent tissue that focuses light on the retina, allowing high‐resolution vision. The loss of lens transparency causes an eye defect termed cataract, which is the leading cause of blindness worldwide. Half of the U.S. population above age 80 is affected by cataracts, for which surgical treatment is the principal therapeutic intervention. In 2014, over 3.6 million cataract surgeries were performed in the United States, and over 20 million surgeries were performed globally. Age‐related cataracts are caused by a variety of stress conditions including elevated oxidative stress, increased sugar (diabetes), smoking or environmental insults such as exposure to ultraviolet (UV) radiation. Thus, characterization of biological and environmental factors that affect lens transparency is the first critical step toward the development of new therapies for cataracts. Recent discoveries have revealed a surprising link between proteins that are components of cytoplasmic RNA granules (RGs) and mammalian cataract. RGs like Processing Bodies (PBs) and Stress Granules (SGs) represent specialized cytoplasmic sites for regulation of RNA in the control of gene expression. PBs, the constitutive class of RGs, are RNA‐protein complexes that silence or channel mRNA to decay, and therefore function to regulate the cellular proteome. SGs form as an early reaction to cellular stress, specifically allowing the translation of proteins that function in homeostasis. To investigate RG function in lens cells, we molecularly characterized two human lens epithelial cell lines (LECs), SRA01/04 and HLE‐B3. These previously uncharacterized cell lines can serve as valuable in vitro tools for investigating the intricate molecular basis for human cataractogenesis. It is important in cell culture studies to validate the origin and identity of the cell line. Therefore, our first step was to confirm that the cell lines were of human origin. Analyzing genome‐specific Short Tandem Repeats by PCR, we authenticated that the LEC lines are indeed human‐derived. We next analyzed gene expression in these LECs by high throughput Illumina microarray analysis to investigate the extent of their retention of lens‐like character. The microarray data demonstrates that both LECs retain expression of genes that are expressed and/or enriched in normal lens epithelial cells. Using a bioinformatics tool “iSyTE” developed to predict genes implicated in lens biology and cataract, we further validated the expression of cataract associated genes in these LECs using RT‐PCR. Furthermore, we find that both LECs support formation of RGs, and exhibit formation of SGs under various stress conditions such as chemical and UV‐stress. Thus, these studies present the first cellular models for investigating the molecular biology of UV‐radiation exposure in lens epithelial cells, and therefore represent a new resource for study of factors that are linked to age‐associated cataract in humans.Support or Funding InformationFunding was provided by the 2015 Delaware Governor's Bioscience Fellowship to Joshua Barton, and University of Delaware Startup funds as well as a grant from The Pew Charitable Trusts awarded to Salil Lachke.

Expression and its significance of aquaporin0 and 1 in lens with age-related cataract

Background Age-related cataract is a common cause of blindness.However, its cause and pathogenic mechanism have not been fully understood.Recent studies revealed that aquaporin 1 (AQP1) and AQP0 are closely related to the pathogenesis of cataract. Objective This study was to investigate the differential distribution and expression of AQP0 and AQP1 in lenses with age-related cataract and explore its effect on pathogenesis of age-related cataract. Methods Seventeen anterior capsular membrane samples and nucleus samples of lenses were collected from age-related cataract patients during the small incision nonphacoemulsification cataract extraction, and 6 normal lens samples were obtained from health donors in the First Affiliated Hospital of Fujian Medical University.The expression and distribution of AQP1 and AQP0 in the lenses were detected by immunohistochemistry, and the relative expression levels of AQP1 and AQP0 proteins in the lenses were assayed by using Western blot assay.This study protocol was approved by Ethic Committee of this hospital, and written informed consent was obtained from each patient. Results Immunohistochemistry showed that in the normal lenses, AQP1 expressed mainly in LECs; while AQP0 primarily expressed in fiber cells of the lens cortex and nucleus.The relative expression levels of AQP1 and AQP0 in the lenses with age-related cataract (absorbance) were 0.223±0.008 and 0.118±0.015, which were significantly lower than 0.246±0.007 and 0.149±0.007 in the normal lenses (t=-4.508, -3.291, both at P<0.01). Western blot revealed that the relative expression levels of AQP1 and AQP0 in the lenses with age-related cataract (absorbance) were 0.663±0.012 and 0.599±0.015, which were significantly reduced in comparison with 0.844±0.041 and 0.955±0.064 in the normal lenses (t=-7.492, P<0.05; t=-9.570, P<0.01). Conclusions AQP1 and AQP0 distribute in different sites of lenses.The expressions of AQP1 and AQP0 are obviously down-regulated in lenses with age-related cataract, suggesting that AQP1 and AQP0 probably play different roles in the pathogenesis of age-related cataract. Key words: Aquaporin/metabolism; Lens, crystalline/metabolism; Eye proteins/metabolism; Water/metabolism; Cataract, age-related/pathogenesis; Humans; Cell membrane permeability/physiology

Quantitative Scheimpflug camera study of the lens density in healthy Turkish population.

The purpose of this study is to evaluate the association between the age and the density of anterior, posterior, and nuclear parts of lens and detect the normal lens density (LD) values in different age groups. 370 eyes of 370 voluntary healthy Turkish subjects were included. These were divided in four groups according to their age. These were Group 1 (51-65years), Group 2 (41-50years), Group 3 (31-40years) and Group 4 (20-30years). LD values of anterior, nuclear, posterior, and total (average) were measured using Pentacam HR Scheimpflug camera system. Both eyes of each subject were scanned twice in automatic release and 50-picture3-dimensional (3D) scan modes by one experienced observer. 207 of the participants were females and 163 were males, with a mean age of 40.57±13.06years. The differences between groups in terms of nuclear area, posterior area, and average LD measurements were significant but anterior area was not. The nuclear area and average LD had high positive correlation with age (r=0.72, p<0.001, r=0.63, p<0.001, respectively); posterior area LD had a weak negative correlation with age (r=-0.16, p=0.001). The mean LD values between female and male subjects showed no significant difference. The anterior region of the lens has the highest and nuclear area has the lowest value of LD in young people. In older people, the anterior area did not show a significant change, nuclear and total areas showed a significant increase but posterior area showed a significant decrease.

AGEs in human lens capsule promote the TGFβ2-mediated EMT of lens epithelial cells: implications for age-associated fibrosis.

SummaryProteins in basement membrane (BM) are long‐lived and accumulate chemical modifications during aging; advanced glycation endproduct (AGE) formation is one such modification. The human lens capsule is a BM secreted by lens epithelial cells. In this study, we have investigated the effect of aging and cataracts on the AGE levels in the human lens capsule and determined their role in the epithelial‐to‐mesenchymal transition (EMT) of lens epithelial cells. EMT occurs during posterior capsule opacification (PCO), also known as secondary cataract formation. We found age‐dependent increases in several AGEs and significantly higher levels in cataractous lens capsules than in normal lens capsules measured by LC‐MS/MS. The TGFβ2‐mediated upregulation of the mRNA levels (by qPCR) of EMT‐associated proteins was significantly enhanced in cells cultured on AGE‐modified BM and human lens capsule compared with those on unmodified proteins. Such responses were also observed for TGFβ1. In the human capsular bag model of PCO, the AGE content of the capsule proteins was correlated with the synthesis of TGFβ2‐mediated α‐smooth muscle actin (αSMA). Taken together, our data imply that AGEs in the lens capsule promote the TGFβ2‐mediated fibrosis of lens epithelial cells during PCO and suggest that AGEs in BMs could have a broader role in aging and diabetes‐associated fibrosis.

Elucidating the Coordination Features Associated to Copper and Zinc Induced Aggregation of Human γ-D Crystallin

Cataract is the leading cause of blindness in the world. The monomeric all -sheet Human γD (HγD) crystallin is an abundant crystallin in the core of the lens and its non-amyloid aggregation is associated with cataracts. A variety of experimental and etiological studies implicate metals as a potential etiological agent for cataract. Copper and zinc concentrations in the cataractous lenses are increased significantly, as compared to normal lenses. Micromolar concentrations of Cu(II) and Zn(II) ions specifically induce the aggregation of HγD crystallin. The mechanism for metal-induced aggregation likely involves interaction of the metal ions with the protein, leading to partial unfolding and formation of high molecular weight light scattering aggregates. In this work, we study the coordination features associated to the metal-induced aggregation of HγD crystallin and probe it by site-directed mutagenesis. Several His and Cys residues have been identified as putative metal anchoring sites. The effects of mutations at these residues in the copper- and zinc- induced aggregation of HγD crystallin have been evaluated. A specific interaction of Zn(II) ions with the N-terminal domain that is key for zinc-induced aggregation has been identified. On the other hand, circular dichroism and electron paramagnetic resonance data show that Cu(II) ions can bind at more than one site in the protein. These results will help elucidate the mechanism by which copper and zinc ions induce the aggregation of HγD crystallin, as it relates to cataracts disease. This research has been supported by: MIT-Seed Funds, Conacyt (#221134), NIH EY015834, and Fulbright-Garcia Robles fellowship to L.Q.

Effect of NF-κB p65 antisense oligodeoxynucleotide on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2.

To study the inhibition of nuclear factor kappa-B p65 (NF-κB p65) antisense oligodeoxynucleotide (ASODN) on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2 (TGF-β2) in vitro. NF-κB p65 ASODN and NF-κB p65 missense oligodeoxynucleotide (MSODN) were designed and synthesized. Human lens epithelial cell line (HLE B-3) cells were prepared for study and divided into 7 groups. Control group was HLE B-3 cells cultured in vitro in dulbecco's modified eagle medium (DMEM). T1, T2, and T3 group were HLE B-3 cells cultured in vitro in DMEM with 10 ng/mL TGF-β2 for 6h, 12h, 24h respectively. A+T group was HLE B-3 cells cultured with 10 ng/mL TGF-β2 for 24h after transfected by NF-κB p65 ASODN for 24h. M+T group was HLE B-3 cells cultured with 10 ng/mL TGF-β2 for 24h after transfected by NF-κB p65 MSODN for 24h. The negative control group was HLE B-3 cells cultured with 10 ng/mL TGF-β2 for 24h after cultured with transfer agent (HiPerFect) for 24h. Cell morphology was observed at different time points using an inverted microscope. The expression of NF-κB p65 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR), and the expression of α-smooth muscle actin (α-SMA) protein was assayed with ELISA. With the TGF-β2 stimulation prolongation, the expression of NF-κB p65 mRNA and α-SMA protein increased in T1, T2, T3 groups compared with the control group, and the difference was statistically significant (P<0.05). NF-κB p65 ASODN lowered the expression of NF-κB p65 mRNA and α-SMA protein induced by TGF-β2. NF-κB p65 MSODN and HiPerFect did not lower the expression of NF-κB p65 mRNA and α-SMA protein induced by TGF-β2. The difference between control group and A+T group was not statistically significant (P>0.05), but the difference among A+T group and other groups was statistically significant (P<0.05). NF-κB p65 ASODN could lower the expression of NF-κB p65 mRNA and α-SMA protein induced by TGF-β2, and antagonized TGF-β2-induced transdifferentiation of HLE B-3 in vitro. NF-κB p65 ASODN could be used as a new biological therapeutic target of posterior capsular opacification.

Biological Association and Expressions of NOS3 &amp;amp; SOD2 in Non-Diabetic Senile Cataractogenesis

Aim: To evaluate the functional relationship between the nitric oxide synthase (NOS) and superoxide dismutase (SOD) enzymes in the pathogenesis of human senile cataract lenses of non-diabetic patients. Methods: Total solubilized proteins from human cataract lens were compared with normal lens (control) by 2-Dimenstional gel electrophoresis (2-DE). Proteins with different abundances were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Western blot analysis was used to verify the changes in expression of NOS3 and SOD2. A further functional association of NOS3 with SOD2 and other proteins was seen by STRING 8.3 databases. Results: In the 2-DE maps, the cataract and normal lens proteins migrated in the region of pH 3 - 10 with a relative molecular weight of 20 - 130 kDa. Approximately two protein spots with differential intensity were detected as NOS3 and SOD2 using MALDI-TOF-MS. Western blot analysis showed high expression of NOS3 in cataract and SOD2 in normal lens samples. String interaction network revealed strong interactions between NOS3 and SOD2 at high confidence score, which is helpful in characterization of functional abnormalities that may be a causative factor in the pathogenesis of cataract. Conclusion: This study will offer new avenues for mechanistic evaluation and future prevention of cataractogensis. However, large scale studies will be required to evaluate the effect of this interaction on the clinical outcome in human cataract.

Flame propagation behaviors of nano- and micro-scale PMMA dust explosions

Flame propagation behaviors of nano- and micro-polymethyl methacrylate (PMMA) dust explosions were experimentally studied in the open-space dust explosion apparatus. High-speed photography with normal and microscopic lenses were used to record the particle combustion behaviors and flame microstructures. Simple physical models were developed to explore the flame propagation mechanisms. High-speed photographs showed two distinct flame propagation behaviors of nano- and micro-PMMA dust explosions. For nano-particles, flame was characterized by a regular spherical shape and spatially continuous combustion structure combined with a number of luminous spot flames. The flame propagation mechanism was similar to that of a premixed gas flame coupled with solid surface combustion of the agglomerates. In comparison, for micro-particles, flame was characterized by clusters of flames and the irregular flame front, which was inferred to be composed of the diffusion flame accompanying the local premixed flame. It was indicated that smaller particles maintained the leading part of the propagating flame and governed the combustion process of PMMA dust clouds. Increasing the mass densities from 105 g/m3 to 217 g/m3 for 100 nm PMMA particles, and from 72 g/m3 to 170 g/m3 for 30 μm PMMA particles, the flame luminous intensity, scale and the average propagation velocity were enhanced. Besides, the flame front became more irregular for 30 μm PMMA dust clouds.