Bim is responsible for the inherent sensitivity of the developing retinal vasculature to hyperoxia

AbstractApoptosis plays an important role in development and remodeling of vasculature during organogenesis. Coordinated branching and remodeling of the retinal vascular tree is essential for normal retinal function. Bcl-2 family members, such as bim not only influence apoptosis, but also cell adhesive and migratory properties essential during vascular development. Here we examined the impact of bim deficiency on postnatal retinal vascularization, as well as retinal neovascularization during oxygen-induced ischemic retinopathy (OIR) and laser-induced choroidal neovascularization. Loss of bim expression was associated with increased retinal vascular density in mature animals. This was mainly attributed to increased numbers of pericytes and endothelial cells. However, the initial spread of the superficial layer of retinal vasculature and, the appearance and density of the tip cells were similar in bim+/+ and bim−/− mice. In addition, hyaloid vessel regression was attenuated in the absence of bim. Furthermore, in the absence of bim retinal vessel obliteration and neovascularization did not occur during OIR. Instead, normal inner retinal vascularization proceeded independent of changes in oxygen levels. In contrast, choroidal neovascularization occurred equally well in bim+/+ and bim−/− mice. Together our data suggest bim expression may be responsible for the inherent sensitivity of the developing retinal vasculature to changes in oxygen levels, and promotes vessel obliteration in response to hyperoxia

Noninvasive temporal detection of early retinal vascular changes during diabetes

Diabetes associated complications, including diabetic retinopathy and loss of vision, are major health concerns. Detecting early retinal vascular changes during diabetes is not well documented, and only few studies have addressed this domain. The purpose of this study was to noninvasively evaluate temporal changes in retinal vasculature at very early stages of diabetes using fundus images from preclinical models of diabetes.Non-diabetic and Akita/+ male mice with different duration of diabetes were subjected to fundus imaging using a Micron III imaging system. The images were obtained from 4 weeks- (onset of diabetes), 8 weeks-, 16 weeks-, and 24 weeks-old male Akita/+ and non-diabetic mice. In total 104 fundus images were subjected to analysis for various feature extractions. A combination of Canny Edge Detector and Angiogenesis Analyzer plug-ins in ImageJ were utilized to quantify various retinal vascular changes in fundus images. Statistical analyses were conducted to determine significant differences in the various extracted features from fundus images of diabetic and non-diabetic animals. Our novel image analysis method led to extraction of over 20 features. These results indicated that some of these features were significantly changed with a short duration of diabetes, and others remained the same but changed after longer duration of diabetes. These patterns likely distinguish acute (protective) and chronic (damaging) associated changes with diabetes. We show that with a combination of various plugging one can extract over 20 features from retinal vasculature fundus images. These features change during diabetes, thus allowing the quantification of quality of retinal vascular architecture as biomarkers for disease progression. In addition, our method was able to identify unique differences among diabetic mice with different duration of diabetes. The ability to noninvasively detect temporal retinal vascular changes during diabetes could lead to identification of specific markers important in the development and progression of diabetes mediated-microvascular changes, evaluation of therapeutic interventions, and eventual reversal of these changes in order to stop or delay disease progression

Adenosine Receptors Expression in Human Retina and Choroid with Age-related Macular Degeneration

Purpose: Adenosine signaling modulates ocular inflammatory processes, and its antagonism mitigates neovascularization in both newborns and preclinical models of ocular neovascularization including age-related macular degeneration (AMD). The adenosine receptor expression patterns have not been well characterized in the human retina and choroid. Methods: Here we examined the expression of adenosine receptor subtypes within the retina and choroid of human donor eyes with and without AMD. Antibodies specifically targeting adenosine receptor subtypes A1, A2A, A2B, and A3 were used to assess their expression patterns. Quantitative real-time PCR analysis was used to confirm gene expression of these receptors within the normal human retina and choroid. Results: We found that all four receptor subtypes were expressed in several layers of the retina, and within the retinal pigment epithelium and choroid. The expression of A1 receptors was more prominent in the inner and outer plexiform layers, where microglia normally reside, and supported by RNA expression in the retina. A2A and A2B showed similar expression patterns with prominent expression in the vasculature and retinal pigment epithelium. No dramatic differences in expression of these receptors were observed in eyes from patients with dry or wet AMD compared to control, with the exception A3 receptors. Eyes with dry AMD lost expression of A3 in the photoreceptor outer segments compared with eyes from control or wet AMD. Conclusion: The ocular presence of adenosine receptors is consistent with their proposed role in modulation of inflammation in both the retina and choroid, and their potential targeting for AMD treatment

Experimental method for optimizing the molding conditions of hot-pressed briquette

In response to the problems of low strength and high permeability of coal materials in the current physical simulation test of coal mine gas dynamics, a set of experimental research methods for optimizing the molding conditions of hot-pressed briquette. Firstly, a hot-pressed briquette test system was independently established and the advantages and future improvement directions of the test system were summarized. At the same time, based on the Horsfield dense stacking theory, the optimal preparation plan for coal briquette materials was formulated. Finally, a molding condition optimization method was developed that combines the Markov distance measurement method and the golden section method. To verify the effectiveness of the experimental method, the secondary carbonization experiments of briquette were conducted under different molding pressure conditions by controlling the molding temperature to 311.8 ℃, heating rate to 5 ℃/min, and holding time to 5.3 h. The response characteristics of the microstructure, physical and mechanical properties, and permeability characteristics of the hot-pressed briquette under different molding pressure conditions were studied. The results show that with the increase of molding pressure, the total porosity gradually decreases, and the uniaxial compressive strength shows a trend of first increasing and then decreasing. The main forms of failure are block spalling and longitudinal fracture. The initial permeability shows a trend of first decreasing and then increasing, while the minimum permeability shows a trend of first decreasing, then increasing and then decreasing. Using the specific values of each molding condition as the test points, and the key parameters of hot-pressed coal and raw coal as the evaluation parameters, a sample matrix was constructed to calculate the Mahalanobis distance between hot-pressed briquette and raw coal under each molding condition, and then optimize the experimental interval using the golden section method. The optimized final molding pressure is 80 MPa. Under these molding conditions, the density, uniaxial compressive strength, and initial permeability of the hot-pressed briquette produced are 1.137 g/cm3, 12.21 MPa, and 1.32 × 10−15 m2, respectively. They are highly similar to the 1.132 g/cm3, 12.83 MPa, and 1.08 × 10−15 m2 of the raw coal, achieving the goal of improving the strength of the briquette and reducing the permeability of the briquette

Micro-damage model of gas-bearing coal under load and instability identification criteria

The distribution of pores and skeletons within coal reservoirs significantly affects the migration of gases and the occurrence of gas dynamic disasters. To further explore the micro-damage mechanisms in gas-containing coal, a detailed study of the micro-damage process in gas-containing coal was conducted. Atomic force microscopy was employed to conduct in-situ tests on the surfaces of protruding and non-protruding coal samples before and after loading. The results indicate that the surface structure of the coal samples changes after loading, with a reduction in closed pore diameter, damage to some pores, and a tendency for connectivity between adjacent closed pores. Before loading, the pores in coal samples exhibit irregular distribution, while after loading, pore connectivity increases, and the number of open pore throats slightly increases. Loading leads to a reduction in the modulus of coal skeleton in protruding coal samples due to pore connectivity, while non-protruding coal samples experience internal structure compaction, resulting in a slight increase in elastic modulus due to their higher strength. Micro-damage types and concepts in coal were defined, and the stress distribution characteristics around coal pores and the coal skeleton were analyzed, revealing the micro-damage mechanisms in gas-containing coal under different conditions. Simultaneously, the factors influencing the closed-cell micro-gas explosion were discussed. The stress at the end of a slender elliptical hole is greater along the hole wall, making it more susceptible to closed-cell micro-gas explosions. Two forms of occurrence of open-pore micro-damage were described, revealing the constraining effect of the "bottleneck effect" on micro-damage. Inherent fractures were identified as the weak link in the coal skeleton, and the evolution of their rupture was analyzed. Utilizing theories such as linear elastic fracture mechanics, elastic-plastic mechanics, and permeation mechanics, criteria for detecting pore damage and coal instability under stress disturbances were established. The micro-damage characteristics of gas-containing coal and the mechanisms inducing coal and gas outbursts were summarized, and the research direction of coal and gas outburst was prospected

Biochemistry and Molecular Biology b2-Adrenergic Receptor Antagonism Attenuates CNV Through Inhibition of VEGF and IL-6 Expression

Citation: Lavine JA, Farnoodian M, Wang S, et al. b2-adrenergic receptor antagonism attenuates CNV through inhibition of VEGF and IL-6 expression. Invest Ophthalmol Vis Sci. 2017;58:299-308. DOI:10.1167/ iovs.16-20204 PURPOSE. The role of b-adrenergic receptor (AR) signaling in neovascular ocular diseases has recently emerged. We have previously reported that intraperitoneal propranolol inhibits choroidal neovascularization (CNV) in vivo and b2-AR blockade reduces vascular endothelial growth factor (VEGF) expression in mouse retinal pigment epithelium and choroidal endothelial cells in culture. Here we tested the hypothesis that the b2-AR regulates CNV through modulation of VEGF and inflammatory cytokine expression. METHODS. Mice were subjected to laser burns, inducing CNV, and were treated with an intravitreal b2-AR antagonist. After 3 and 5 days, total eye interleukin-6 (IL-6) and VEGF protein levels were measured, respectively. After 14 days, CNV was measured on choroidalscleral flatmounts. The effects of b-AR signaling on VEGF and IL-6 expression were investigated in various mouse retinal and human RPE cells by using specific b-AR agonists and antagonists. RESULTS. b2-Adrenergic receptor signaling increased Vegf mRNA expression by approximately 3-to 4-fold in mouse retinal microglia and pericytes in culture. b2-Adrenergic receptor signaling upregulated IL-6 mRNA expression between 10-and 60-fold in mouse retinal microglia, pericytes, RPE, and choroidal endothelial cells in culture. Intravitreal injection of b2-AR antagonist ICI 118,551 reduced CNV by 35% and decreased IL-6 protein levels by approximately 50%. In primary human RPE cells, b2-AR activation also stimulated VEGF and IL-6 mRNA expression by 2-and 10-fold, respectively. CONCLUSIONS. Anti-VEGF therapy for CNV is highly effective; however, some patients are resistant to therapy while others undergo repeated, frequent treatments. b2-Adrenergic receptor signaling is a potential therapeutic target because of its angiogenic and inflammatory properties

Bim Expression Promotes the Clearance of Mononuclear Phagocytes during Choroidal Neovascularization, Mitigating Scar Formation in Mice

Inflammation is increasingly recognized as an important modulator in the pathogenesis of neovascular age-related macular degeneration (nAMD). Although significant progress has been made in delineating the pathways that contribute to the recruitment of inflammatory cells and their contribution to nAMD, we know little about what drives the resolution of these inflammatory responses. Gaining a better understanding of how immune cells are cleared in the choroid will give a novel insight into how sustained inflammation could influence the pathogenesis of nAMD. The pro-apoptotic Bcl-2 family member Bim is a master regulator of immune cell homeostasis. In its absence, immune cell lifespan and numbers increase. Most therapeutic regimes that squelch inflammation do so by enhancing immune cell apoptosis through enhanced Bim expression and activity. To test the hypothesis that Bim expression tempers inflammation during the pathogenesis of nAMD, we used the mouse laser-induced choroidal neovascularization (CNV) model in which inflammation acts as a facilitator of CNV. Here, we showed minimal to no change in the recruitment of F4/80-, CD80-, CD11b-, and Iba1-positive myeloid-derived mononuclear phagocytes to the site of laser photocoagulation in the absence of Bim expression. However, the resolution of these cells from the choroid of Bim-deficient (Bim -/-) mice was significantly diminished following laser photocoagulation. With time, we noted increased scar formation, demonstrated by collagen I staining, in Bim -/- mice with no change in the resolution of neovascularization compared to wild-type littermates. We also noted that mice lacking Bim expression in mononuclear phagocytes (BimFlox/Flox; Lyz2-Cre (BimMP) mice) had delayed resolution of F4/80-, CD80-, CD11b-, and Iba1-positive cells, while those lacking Bim expression in endothelial cells (BimFlox/Flox; Cad5-Cre (BimEC) mice) had delayed resolution of only CD11b- and Iba1-positive cells. Both BimMP and BimEC mice demonstrated increased scar formation, albeit to differing degrees. Thus, our studies show that resolving inflammation plays an important role in moderating scar formation in nAMD, and it is impacted by Bim expression in both the endothelium and mononuclear phagocyte lineages

An Empirical Analysis of the Factors Affecting the Adoption and Diffusion of GBTS in the Construction Market

This study focuses on better development of green buildings. The key to the sustainable development of the construction industry is to popularize and promote the spread of green building technologies (GBTS) in the construction market. This study integrates the technology acceptance model (TAM) and the innovation diffusion theory (IDT) to analyze and construct the theoretical model of developers’ GBTS adoption behavior from three dimensions, including the individual factor, product factor and interface factor. This paper discusses the mechanism of GBTS adoption and diffusion in the construction market. The data are collected by questionnaire, and the structural equation model (SEM) is used for empirical analysis. The results show that the developers’ perceived usefulness (PU) and perceived ease of use (PEOU) of GBTS, developers’ innovativeness and sense of community at the individual level, competitive advantage at the product level, as well as government structural guarantees and relevant stakeholders at the interface level have a significant positive impact on the adoption of GBTS by developers. It is proved that the model can explain the basic path of GBTS adoption by developers, and suggestions to promote the adoption and diffusion of GBTS in China are put forward