The Efficacy of Hyperbaric Oxygen for the Treatment of Experimental Uveitis Induced in Rabbits

Objective: The aim of the present study was to examine the efficacy of hyperbaric oxygen (HBO) therapy in the treatment of experimental uveitis induced in rabbits. It was hypothesized that HBO therapy improves the regression of experimental uveitis induced in rabbits. Research design and methods: An experimental animal study was conducted on 48 rabbits (48 right eyes of these rabbits) to evaluate the effects of HBO therapy on endotoxin-induced acute anterior uveitis in rabbits. To induce acute anterior uveitis, Salmonella typhimurium lipopolysaccharide endotoxin (LPS) was intravitreally injected into the right eyes of the rabbits. The animals were randomly assigned to five groups. No treatment was given to the rabbits in Group A. Prednisolone acetate was topically administered to the rabbits in Group B. Methylprednisolone acetate was administered by anterior subtenon injection to the rabbits in Group C four hours after LPS application. HBO therapy was administered to the rabbits in Group D. Both HBO therapy and anterior subtenon injection of methylprednisolone therapy were administered to the rabbits in Group E. To compare the effects of the different therapies on the progression of endotoxin-induced uveitis, examinations including clinical scoring of anterior uveitis, microscopic examination of aspirated aqueous humor for inflammatory responses, and aqueous protein level assessment were performed once a day after LPS injection. Results: There was a statistically significant difference between the control group (Group A) and other groups (Groups B–E) with respect to the number of inflammatory cells and protein levels in the aqueous one and three days after LPS injection (p < 0.05), indicating that the treatments resulted in less inflammation in Groups B–E compared to Group A. Moreover, there was no statistically significant difference between Groups B and C, Groups B and D, Groups B and E, Groups C and D, and Groups C and E with regard to the number of inflammatory cells in the aqueous at Day 1 after LPS injection (p > 0.05). In addition, Groups B and C and Groups B and D were comparable with regard to cell counts at Day 3 (p > 0.05), showing that HBO was comparable to corticosteroids in reducing inflammation. The differences between Groups B and E and Groups C and E were significant with regard to aqueous cell counts at Day 3 (p < 0.05), showing that HBO plus steroid was more effective than steroids alone. Conclusion: The intensity of ocular inflammation in the group receiving HBO therapy combined with anterior subtenon injection of methylprednisolone therapy was lower than in the other groups. We also demonstrated that HBO therapy was an effective therapeutic modality for the treatment of experimental uveitis induced in rabbits with an efficacy comparable to that of corticosteroids. Moreover, HBO plus steroid was superior to steroids alone in reducing inflammation.     

脉络膜上腔不同给药途径治疗葡萄膜炎的实验研究

目的 比较脉络膜上腔给药与其他途径局部给药治疗葡萄膜炎的疗效。方法 36只青紫蓝兔先制作实验性葡萄膜炎模型，分别经脉络膜上腔给药、球内注射、筋膜下注射曲安奈德1mg后，定期测定房水蛋白量、细胞数并行炎症评分、眼底造影及组织病理研究。结果 3种给药途径疗效优劣依次为脉络膜上腔给药、玻璃体内给药、筋膜下给药。结论 脉络膜上腔给药安全有效，可作为一种新的葡萄膜炎治疗手段。     

Effects of Kakkon-to and Sairei-to on experimental elevation of aqueous flare in pigmented rabbits.

PURPOSE: To evaluate the possible inhibitory effects of Kakkon-to and Sairei-to, traditional Sino-Japanese herbal medicines, on experimental aqueous flare elevation in pigmented rabbits. METHODS: Anterior uveitis was induced either by an application of prostaglandin E2 (PGE2), 10 microg/mL, to the cornea, or an intravenous injection of lipopolysaccharides (LPS), 0.5 microg/kg, in an ear vein. Dose dependency of experimental uveitis induced by LPS (0.1, 0.25, 0.5, or 1.0 microg/kg) was also determined. For pretreatment, about 150 g/day of food containing Kakkon-to (1% w/w) or Sairei-to (0.6% or 2% w/w) was given to two groups of animals for 5 days before experimental uveitis was induced. A third group of animals underwent pretreatment with betamethasone, 130 microg/kg, injection into an ear vein 4 hours before experimental uveitis was induced. A fourth group of rabbits with no herbal medicine or betamethasone pretreatment served as controls. Aqueous flare was measured using a laser flare-cell meter. Aqueous flare intensity was expressed as the area under the curve (AUC) in arbitrary units. RESULTS: The increase in aqueous flare induced by LPS was dose-dependent. The AUC of PGE2 (10 microg/mL) and LPS (0.5 microg/mL) induced aqueous flare elevations were 1,119 and 4,950 arbitrary units, respectively. Kakkon-to (AUC, 1,055) and Sairei-to (AUC, 965) did not inhibit the aqueous flare elevation induced by PGE2. Beta-methasone did inhibit the elevation (AUC, 271). Kakkon-to (AUC, 4,495) did not suppress the aqueous flare elevation induced by LPS. Both 0.6% and 2% Sairei-to (AUC, 2,478, and 978) and beta-methasone (AUC, 443) did suppress the aqueous flare elevation induced by LPS significantly (P < .05). CONCLUSION: Sairei-to could have an inhibitory effect on experimental anterior uveitis induced by LPS.     

Encapsulated triamcinolone cyst after subtenon injection

The authors report the formation of a subtenon cyst following subtenon triamcinolone injection for chronic recurrent uveitis in Behçet’s disease. The steroid was inadvertently placed superficially in the anterior subtenon space and was encapsulated eventually to present as a subtenon cyst. The episode of uveitis recurrence, which did not regress, required treatment with a repeat posterior subtenon injection 2 months later. The cyst was removed 4.5 months after the initial subtenon injection, when the patient underwent a trabeculectomy for refractory glaucoma. Histological examination of the cyst revealed a fibrous encapsulated cavity filled with small birefringent crystals, consistent with an encapsulated triamcinolone collection. Fibrous encapsulation of triamcinolone crystals can arise after a superficially placed anterior subtenon injection and this may impede the absorption of the corticosteroid and hamper its effectiveness in treating ocular inflammatory diseases.     

Evaluation of Neopterin Levels in an Endotoxin-Induced Experimental Uveitis Model

Background: In this study, NP levels of intraocular fluids and serum were analyzed in the endotoxin-induced uveitis model. Methods: Intravitreal injection of 0.05?ml E. Coli lipopolysaccharide (IVT LPS) has been performed into the right eyes of 14 rabbits. Animals were divided into two groups. Group 1 (n?=?7) sacrificed 24 hours after the intravitreal injection, and group 2 (n?=?7) 72 hours after the intravitreal injection. Aqueous fluid, vitreous and serum samples were drawn at the time of sacrifice; histopathological changes were also examined. Results: Significantly elevated aqueous and vitreous NP levels were observed in IVT LPS injected eyes. In IVT LPS injected eyes, aqueous NP levels showed a significant decrease in 72 hours (p?p?). Both aqueous and vitreous protein concentrations were significantly higher in IVT LPS injected eyes. Histopathologic evaluation revealed that, in the first 24 hours, inflammation was significant in choroid and ciliary body. Retinal histopathological changes were observed mainly at 72 hours. Conclusions: This study demonstrates that neopterin levels correlate with early intraocular inflammatory response in an endotoxin-induced uveitis model.     

葡萄膜炎并发白内障术中前房注射长效糖皮质激素的疗效

目的：研究术中前房注射长效糖皮质激素是否能提高葡萄膜炎并发白内障手术的疗效,以期为临床诊疗提供参考。方法：分析2013-01/2015-01我院收治的接受手术治疗的葡萄膜炎并发白内障患者68例71眼的临床资料。依据治疗方法的不同分为观察组(术中前房注射曲安奈德,35例37眼)及对照组(术中不注射曲安奈德,33例34眼)。对比两组患者的疗效。结果：观察组术后1mo矫正视力>0.5比例显著高于对照组,差异有统计学意义(χ2=4.094,P=0.043)。两组术前及术后眼压比较均无统计学意义(P>0.05)。观察组患者术后前房炎症反应低于对照组,差异具有统计学意义(χ2=15.900,P=0.001);术后1wk,观察组房水内SOD水平显著高于对照组,差异有统计学意义(P<0.05),而MDA及TNF-α水平显著低于对照组水平,差异有统计学意义( P<0.05)。观察组术后1a累积复发率显著低于对照组,差异有统计学意义(14%,5/35 vs 3%,11/33; Log-rank χ2=4.004,P=0.045)。结论：术中前房注射曲安奈德可显著提高葡萄膜炎并发白内障患者手术的疗效。     

Etanercept treatment in the endotoxin-induced uveitis of rats

This study was conducted to investigate therapeutic value of a soluble tumor necrosis factor-alpha (TNF-alpha) receptor, etanercept, in a rat model of endotoxin-induced uveitis (EIU). Forty-two inbred male Lewis rats were divided into seven equal groups. 200 microg of Escherichia coli 055:B55 lipopolysaccharide (LPS) was injected in one hind footpad of the Groups 2, 3, 4, 5, 6, and 7 rats. Group 5, 6, and 7 rats also received subcutaneous etanercept 24 hr prior to LPS injection at a dose of 0.4 mg kg(-1). Group 1 rats were used as controls. Eight, 24, and 48 hr after treatment clinical uveitis scores (miosis, iris hyperemia, and hypopyon) were assessed by a masked observer and the rats were euthanized. Neutrophil leukocytes, CD8+, CD4+, and CD45RO+ cells in the anterior uveal tissue were counted either after hematoxylin-eosin or monoclonal antibody staining. TNF-alpha levels were also measured in the aqueous humor samples by an ELISA method. Etanercept treatment significantly improved clinical uveitis scores at all examination points compared to the LPS injected animals. The improvement was almost complete expect for the miosis score, since no significant difference was detected between the controls and LPS + Etanercept treated animals at all examination points. Cell counts were also at significantly lower levels in LPS + Etanercept treated animals at all examination points, except for CD8+ and CD45RO+ cell counts at 24 hr examination point. There was no significant difference between the controls and LPS + Etanercept treated animals at all examination points as with CD4+ and CD45RO+ cell counts at 48 hr. Our data showed that etanercept had a definite effect on the treatment of EIU. Further studies should clarify its efficacy on clinical uveitis conditions.     

TLR4-MyD88在大鼠急性前葡萄膜炎虹膜中的表达

目的观察内毒素诱导的大鼠急性前葡萄膜炎（EIU）虹膜组织内Toll样受体4（TLR4）、髓样分化因子88（MyD88）及核因子-κBp65（NF-κB p65）的表达。方法Wistar大鼠50只,随机分为5组,0、12、24、48、72h,每组10只。0h组为正常对照组,其余4组均足垫部注射霍乱弧菌内毒素脂多糖（LPS）200μg,建立EIU动物模型,每隔2h用裂隙灯观察大鼠眼前节炎症反应。通过铺片免疫组织化学染色,检测虹膜睫状体组织内TLR4、MyD88和NF—κB p65的表达,并对虹膜内TLR4^＋和MyD88^＋及NF—κB p65^＋细胞进行计数。结果注射后24～48h大鼠眼前段的炎症反应达到高峰,72h炎症反应逐渐缓解。组织病理学检查表明,虹膜睫状体组织的炎性细胞浸润在24～48h达到高峰,与临床反应结果相符。TLR4在模型鼠虹膜睫状体炎复合体中表达的免疫组织化学检测结果表明,0h组虹膜铺片内无阳性细胞,12h后可见细胞形态大多为类圆形的阳性细胞,48h达高峰,72h阳性细胞数开始减少,各组阳性细胞数总体差异有统计学意义（F=46．79,P〈0．05）。MyD88和NF—κB p65的表达与TLR4的改变趋势相一致（F=54．37,P〈0．05;F=85．32,P〈0．05）。结论内毒素诱导的EIU虹膜内,TLR4及其下游信号传导分子的表达量发生改变,提示TLR4-MyD88依赖传导途径可能参与了EIU的发病.     

Effects of scutellariae radix extract and its components (baicalein, baicalin, and wogonin) on the experimental elevation of aqueous flare in pigmented rabbits

PURPOSE: To evaluate the possible inhibitory effects of hot water extract of Scutellariae radix and its major components (baicalein, baicalin, and wogonin) on experimental elevation of aqueous flare in pigmented rabbits. METHODS: To produce aqueous flare elevation in rabbits, prostaglandin E(2) (PGE(2)), 25 microg/mL, was applied to the cornea with the use of a glass cylinder, or lipopolysaccharides (LPS), 0.5 microg/kg, were injected into an ear vein. Animals were pretreated by the oral administration of 150 g/day of food containing 0.02%, 0.07%, or 0.2% (w/w) extract of Scutellariae radix for 5 days, or by intravenous injection of baicalein, baicalin, or wogonin, 60 microg/kg or 600 microg/kg, 30 minutes before experimental uveitis was induced. Aqueous flare was measured with a laser flare-cell meter. Aqueous flare intensity was expressed as the area under the curve (AUC) in arbitrary units. RESULTS: The AUC of PGE(2)- and LPS-induced aqueous flare elevation was 1,343 and 5,066 arbitrary units, respectively. Pretreatment by oral administration of 0.07% or 0.2% extract of Scutellariae radix did not inhibit PGE(2)-induced aqueous flare elevation (AUC: 1,252 and 1,210, respectively), but it did inhibit LPS-induced aqueous flare elevation (AUC: 2,248 and 1,973, respectively). Pretreatment by intravenous injection of 600 microg/kg of baicalein, baicalin, or wogonin inhibited LPS-induced aqueous flare elevation (AUC: 2,289, 2,163, and 1,509, respectively). Pretreatment with 60 microg/kg of wogonin also inhibited LPS-induced aqueous flare elevation (AUC: 1,980). CONCLUSION: Hot water extract of Scutellariae radix may have an inhibitory effect on experimental anterior uveitis induced by LPS in pigmented rabbits.     

Tobramycin liposomes. Single subconjunctival therapy of pseudomonal keratitis.

The authors compared 24 doses of hourly topical fortified tobramycin (Group A) therapy with a single subconjunctival administration of multivesicular megaliposome-encapsulated tobramycin (Group B) and free subconjunctival tobramycin (Group C) in treating a rabbit model of keratitis caused by Pseudomonas aeruginosa. One cornea each of 50 rabbits was infected with P. aeruginosa for 24 hr. The animals then were divided randomly into five groups of ten each. Groups A, B, and C were treated as described. Group D received liposomes without tobramycin and Group E, hourly balanced salt solution. Significantly fewer Pseudomonas colonies were present in the corneas of all three drug-treated groups (A, B, and C) compared with the two control groups (D and E) at 24 hr (P less than 0.005). Significantly fewer Pseudomonas colonies were present in Groups A and B compared with Group C (P less than 0.02). No significant difference was noted between Groups A and B (P = 0.30). Tobramycin encapsulated in megaliposomes may be useful in treatment of pseudomonal keratitis.     

Endotoxin-induced production of inflammatory mediators by cultured ciliary epithelial cells

Systemic injection of bacterial endotoxin (Lipopolysaccharide, LPS) in experimental animals induces anterior uveitis without major pathological changes in other organs. The present study investigates the effect of LPS on production of inflammatory mediators in cultured bovine pigmented ciliary epithelial cells (CB-cells) by means of radioimmunoassays and bioassays. LPS was found to stimulate CB-cells to secrete prostaglandin E2 and prostacyclin (assayed as its stable metabolite 6-keto-prostaglandin F1a), but not leukotriene B4 or thromboxane A2 (assayed as its stable metabolite thromboxane B2). CB-cells produced membrane-associated interleukin 1-activity in response to LPS, but no tumor necrosis factor-activity was found after challenge of CB-cells with LPS. The direct effect of LPS on production of inflammatory mediators by cells from the anterior uvea could play a role in the pathophysiology of endotoxin-induced uveitis.     

白藜芦醇对内毒素诱导的大鼠葡萄膜炎的治疗作用

目的：探讨眼局部使用白藜芦醇（Res）对内毒素诱导的大鼠葡萄膜炎的治疗效果及相关机制。方法：实验研究。将36 只雄性Wistar大鼠按随机数字表法分为空白组、模型组、0.25% Res治疗组、0.5%Res治疗组、1% Res治疗组和0.1%地塞米松（Dex）治疗组,每组6只。将大肠杆菌细胞壁脂多糖（LPS）溶于无菌的0.9%氯化钠溶液配成1 mg/ml的LPS溶液,并注射入模型组和治疗组大鼠双后足垫（每只注射200 μg）,空白组注射等量0.9%氯化钠溶液。各治疗组于LPS诱导前后分别给予相应浓度的 Res和Dex 10 μl点双眼,每2 小时1 次,注射LPS前后各6 次。空白组和模型组给予等量的0.9%氯化钠溶液点眼。观察大鼠眼部炎症反应、临床表现评分,注射LPS 24 h后测房水肿瘤坏死因子-α （TNF-α）和白细胞介素-6（IL-6）浓度、眼球病理切片HE染色以及免疫组织化学检测虹膜睫状体p38丝裂原活化蛋白激酶（MAPK）和核转录因子-κB（NF-κB）p65 等,探讨与分析Res的治疗效果。采用单因素方差分析、Mann-Whitney U检验进行数据处理。结果：模型组于LPS注射后4 h可见虹膜血管扩张充血,之后炎症反应逐渐加重,24 h时可见前房大量点状渗出、瞳孔区纤维渗出及晶状体前囊膜渗出物沉着,模型组24 h临床评分为4.3 ± 1.2,1% Res组为2.0 ± 0.6,1% Res组葡萄膜炎反应明显轻于模型组（P < 0.05）;1% Res组房水中TNF-α和IL-6的浓度与模型组相比均明显降低（P < 0.05）;1% Res组眼部组织病理学改变较模型组轻（P < 0.05）;1% Res组虹膜睫状体p38 MAPK和NF-κB p65 的核转位阳性细胞率较模型组均明显降低（P < 0.05）。结论：1% Res局部点眼能有效减轻内毒素诱导的大鼠自身免疫性葡萄膜炎的临床症状,其机制可能与抑制MAPK和NF-κB信号通路有关。     

Activated complement in inflamed aqueous humor

Activated complement is an important mediator of inflammation. Radioimmunoassay was used to measure levels of C3a, an activated fragment of C3, in aqueous humor. Additionally, immunoelectrophoresis was performed on aqueous humor to detect Factor B and its conversion product, Bb, as well as C3c, a breakdown product of C3. All six samples of normal aqueous humor had no detectable C3a, C3c, or Factor B. All eight samples of aqueous humor from patients with anterior uveitis had measurable levels of C3a. Factor B and C3c were detected in 3/7 samples of inflamed aqueous humor. Factor B was converted fully to Bb in two of these three samples, suggesting alternative pathway activation of complement. Activated complement fragments are present in the aqueous humor of eyes with anterior uveitis and may help mediate the inflammatory process.     

Inhibitory effect of aminoimidazole carboxamide ribonucleotide (AICAR) on endotoxin-induced uveitis in rats

PURPOSE. To investigate the anti-inflammatory effect of aminoimidazole carboxamide ribonucleotide (AICAR), an analog of adenosine monophosphate (AMP), in endotoxin-induced uveitis (EIU). METHODS. EIU was induced by subcutaneous injection of lipopolysaccharide (LPS) (200 μg) in Lewis rats. AICAR (50 mg/kg, intraperitoneally) was given 6 hours prior and at the same time as LPS injection. Clinical uveitis scores, number of anterior chamber (AC) infiltrating cells, anterior chamber protein concentration, retinal vessel leukocyte adhesion, and protein leakage were measured 24 hours later. Protein levels of C-C chemokine ligand-2 (CCL-2)/monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-α (TNF-α) and intercellular adhesion molecule-1 (ICAM-1) in aqueous humor and retina and nuclear translocation of nuclear factor-κB (NF-κB) in the retina were determined by enzyme-linked immunosorbent assay (ELISA). Both mRNA and protein levels of CD14 in peripheral blood mononuclear cells were also measured. RESULTS. AICAR treatment significantly reduced EIU clinical severity as well as inflammatory cell infiltration and protein concentration in aqueous humor. Similarly, the number of retinal vessel-adherent leukocytes and protein leakage were decreased by AICAR treatment. Protein levels of TNF-α, CCL-2/MCP-1, and ICAM-1 in aqueous humor and CCL-2/MCP-1 and ICAM-1 levels in retina were suppressed with AICAR treatment. AICAR also reduced NF-κB translocation and CD14 expression. CONCLUSIONS. AICAR reduces systemic LPS susceptibility and attenuates intraocular inflammation in a rat EIU model by limiting infiltration of leukocytes, suppressing inflammatory mediators, and inhibiting the NF-κB pathway.     

Monocyte chemotactic activity induced by intravitreal endotoxin

In order to clarify the factors responsible for the cellular infiltrate characteristic of anterior uveitis, the authors have induced inflammation in rabbits by the intravitreal injection of 100 ng of Escherichia coli or Salmonella endotoxin (ET). A 2% concentration of aqueous humor 18 to 24 hr after ET consistently induced monocyte migration as measured in modified Boyden chambers. Activity was significantly greater in these samples than in aqueous after saline injection or 3 hr after endotoxin injection (prior to cellular infiltrate). Using either sephadex G-75 molecular sieve chromatography or a cibacron blue column, the vast majority of migratory activity co-eluted with albumin. Serum albumin, however, at a comparable concentration did not induce migration. Activity was largely heat- and acid-stable and was maximal in the presence of a concentration gradient, indicating that it was chemotactic rather than chemokinetic. A second peak of activity eluted from the G-75 column just prior to a marker with molecular weight of 427 and was also present in eluates from normal aqueous humor. Chloroform:methanol extraction, radioimmunoassay, and high performance liquid chromatography indicated that a small portion of the chemotactic activity could be ascribed to lipid including leukotriene B4. In contrast to the prominence of complement (C5a) derived chemotactic activity resulting from intravenous ET, C5a was not a major contributor to aqueous chemotactic activity subsequent to local ET. These observations demonstrate that leukocyte migration factors in aqueous humor can be characterized and compared. This approach can be used to test the hypothesis that subsets of anterior uveal inflammation might be distinguished on the basis of associated chemotactic factors.     

两性霉素B缓释系统治疗兔烟曲霉菌眼内炎的药效学实验研究

目的探讨两性霉素B缓释系统（AmB—DDS）玻璃体腔植入对烟曲霉菌性眼内炎的疗效、AmB-DDS的药物释放规律及最佳释药量。方法选取40只新西兰白兔作为实验动物。（1）Amb-DDS治疗烟曲霉菌性眼内炎疗效学观察：动物玻璃体腔内注入烟曲霉菌悬液,48h后随机分为5组,A组为空白对照组（6只眼）,B组为空白DDS组（6只眼）,C组为两性霉素B玻璃体腔内注射组（6只眼）,D组为AmB—DDS250μg植入联合玻璃体切除术组（8只眼）,E组为AmB—DDS500μg植入联合玻璃体切除术组（8只眼）。术后不同时间点检测前房闪辉、细胞及玻璃体混浊程度,取玻璃体腔内容物行涂片检查和真菌培养,2个月时取眼球标本行病理学检查;（2）AmB—DDS玻璃体腔内药物浓度检测：H组玻璃体切除术后植入500μg AmB—DDS1个（6只眼）,术后第1、3、7天及2、4、6、8周取玻璃体液,高效液相色谱分析法检测药物浓度。结果A、B组全部发生严重眼内炎,伴眶内感染,组间比较差异无统计学意义（P〉0．05）;C、D、E组炎性反应较A、B组轻,差异有统计学意义（P≤0．005）;E组玻璃体混浊程度较C组轻,7～14d前房反应较C组轻,差异均有统计学意义（P≤0．005）;D组5只眼、E组8只眼治愈,差异有统计学意义（x^2=10．494,P=0．003）。不同时间点取玻璃体腔内容物涂片,所有标本6周内均见菌丝,真菌培养仅A、B组为阳性。病理学检查示治愈眼结构正常,感染未控制眼均萎缩,球壁结构被破坏。H组术后第1天即有释药,药物浓度迅速升高超出有效抑菌浓度,观察期内释药较平稳。结论AmB—DDS玻璃体腔内植入治疗烟曲霉菌性眼内炎安全有效,释药恒定,速率得当;以含药量为500μg的AmB—DDS治疗效果最佳。（中华腰科杂志,2007,43：546-553）     

Characterization of endotoxin-induced C5-derived chemotactic activity in aqueous humor

Although a cellular exudate characterizes acute anterior uveitis, few studies have sought to identify the chemoattractant(s) contributing to this phenomenon. As a model of acute ocular inflammation, the authors have injected rabbits intravenously with endotoxin (Salmonella typhimurium LPS, 2.5 micrograms/kg). In a Boyden chamber assay, aqueous humor drawn 3 hr after LPS (post-LPS aqueous) exhibited chemotactic activity for purified rabbit granulocytes (PMN). "Checkerboard" analysis indicated that chemotaxis, rather than protein-induced chemokinesis, primarily accounted for PMN migration. Aqueous from normal rabbits demonstrated no chemotactic activity. Chemotactic activity was maximal at 3 hr post-LPS (versus 1 or 5 hr). PMN migration exhibited a direct correlation with the concentration of aqueous tested (0.5-5%). Several observations indicated that this chemotactic activity is complement (C5)-derived. It is inhibited by antibodies to C5 but not affected by antibodies to C3. Similar to rabbit C5a, chemotactic activity in post-LPS aqueous was heat stable at 56 degrees C X 30 min, attracted both human and rabbit PMN at similar concentrations and induced release of beta glucuronidase from PMN. In addition, prior incubation of rabbit PMN with partially purified C5a (densensitization) specifically inhibited chemotactic responses to both C5a and post-LPS aqueous without inhibiting responses to another chemoattractant, n-formyl-methionyl-leucyl-phenylalanine. Finally, chemotactic activity from post-LPS aqueous could be recovered from a Sephadex G75 column and eluted similarly to chemotactic activity in zymosan activated rabbit serum or 13,700 D molecular weight marker. The presence of complement-derived chemotactic activity in this model should not be construed as evidence that this activity contributes to the pathogenesis of endotoxin-induced inflammation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Anaphylatoxin levels in human aqueous humor

Radioimmunoassay was used to measure levels of C3a, C4a, and C5a in aqueous humor from 13 normal eyes, 8 noninflamed eyes with a history of surgery or inflammation, and 14 eyes with anterior uveitis. The authors were unable to measure levels of C3a, C4a, or C5a in normal aqueous humor. In noninflamed aqueous humor from eyes with a history of surgery or inflammation, the authors were unable to measure levels of C4a or C5a, but were able to measure low levels of C3a in 3/8 patients. In aqueous humor from eyes with anterior uveitis, the authors were able to measure levels of C3a in all 14 patients, C4a in 9/14 patients, and C5a in 5/14 patients. Patients with severe anterior uveitis had higher levels of C3a than those with moderate anterior uveitis. The higher ratios of anaphylatoxin to protein levels in inflamed aqueous humor, when compared to normal plasma or noninflamed aqueous humor, suggested that complement was being activated by either the classical or alternative pathways in inflamed aqueous humor. Measurable levels of C3a without detectable C4a in five patients with anterior uveitis suggested alternative pathway activation of complement.     

Anaphylatoxin in the aqueous humor of rabbit with endotoxin induced endophthalmitis

Anaphylatoxin-activated complement fragment is an important mediator of inflammation. In this study, trends of anaphylatoxin in clinical and experimental endophthalmitis was evaluated. The level of anaphylatoxin (C3a, C4a and C5a) and protein of human aqueous humor aspirated from patients with senile cataracts before surgery was measured. As a model of acute endophthalmitis, endotoxin-Lipopolysaccharide from E. coli (LPS) was used intravitreously in albino rabbits. The level of anaphylatoxin of aqueous humor was measured by radioimmunoassay. The level of C3a was 44.0 +/- 13.4 ng/ml in aqueous humor of cataract patients. An increase of cell count and protein was demonstrated in the aqueous humor of rabbits 24 hrs after endotoxin injection. The level of C3a increased remarkably with time. It was suggested that C3a in aqueous humor of the rabbit eye with endotoxin-induced endophthalmitis plays an important role in chemotaxis and in mediating the inflammatory process.     

Prospective optical coherence tomographic evaluation of the efficacy of oral and posterior subtenon corticosteroids in patients with intermediate uveitis

Purpose To prospectively evaluate the efficacy of oral corticosteroids and posterior subtenon injection in the treatment of macular edema in patients with intermediate uveitis using optical coherence tomography (OCT).Methods Twenty-two patients with intermediate uveitis were treated with posterior subtenon injection when the disease was unilateral (group A, n=11) or with oral steroids when the disease was bilateral (group B, n=11). Changes in macular thickness from baseline was determined using OCT in both groups at day 0, day 3, day 14, 6 weeks and 12 weeks.Results Statistically significant improvement in Snellen visual acuity in group A was seen at 6 weeks and in group B at 2 weeks. In patients receiving oral corticosteroids, foveal thickness decreased by 63% by day 3. In those treated with posterior subtenon injection, even at day 14 only a 55% reduction of foveal thickness was evident. Spearman’s correlation coefficient for visual acuity and foveal thickness was found to be significant.Conclusion OCT confirms a significantly more rapid decrease in macular edema in patients treated with oral corticosteroids. A short course of oral steroids may be useful in enabling earlier visual recovery in patients treated with posterior subtenon injection for unilateral uveitic macular edema.