Myocilin protein levels in the aqueous humor of the glaucomas in selected canine breeds

Objective To compare aqueous humor myocilin protein levels in dogs with the primary glaucomas to those with the secondary glaucomas, primary cataracts, and diabetic cataracts. Materials and methods Four groups were selected, based on diagnosis by the attending veterinary ophthalmologists and included: primary glaucoma (primary open‐angle glaucoma (POAG) and primary closed angle glaucoma (PCAG); n = 155); secondary glaucoma (n = 94); primary (presumed inherited) cataract (n = 142), and diabetic cataract (n = 83). A total of 474 samples (187 males, 263 females, 24 unreported) with average ages of 117 months for the males and 101 months for the females were analyzed. Myocilin protein was measured using the Coomassie staining and Western blot methods relative to a myocilin control. Results Differences were seen between nonglaucomatous (cataractous) and glaucomatous dogs with myocilin levels in glaucomatous eyes being many times higher than those in the cataractous dogs. Primary glaucomatous dogs were found to have an aqueous humor myocilin protein level of 17.30 ± 1.03 units. Secondary glaucomas had the highest level of myocilin in the aqueous humor with 19.27 ± 1.41 units. Diabetic cataractous dogs had the lowest levels of myocilin reported with 6.60 ± 0.88 (mean ± SEM) units. Normal (cataractous) dogs had a myocilin level in the aqueous humor of 8.05 ± 0.86 units. Conclusion Aqueous humor protein levels were elevated, relative to the myocilin control, in both the primary and secondary glaucoma groups compared to the cataract and diabetic cataract groups. Like in the Beagle POAG, aqueous humor myocilin protein levels are increased. Further studies are indicated to investigate the exact role of the aqueous humor myocilin protein in the genesis in increased IOP in these primary glaucomatous breeds.     

Aqueous humor myocilin protein levels in normal, genetic carriers, and glaucoma Beagles

Objective The gene (myocilin: MYOC) has been attributed to be involved in over 6% of inherited types of human glaucoma, the highest correlation for any gene to date. This study determines myocilin protein levels in the aqueous humor (AH) of normal laboratory quality, genetic carrier (offspring of normal laboratory quality and POAG Beagles), and primary open angle glaucoma (POAG) Beagles. Materials and methods Eighteen dogs were used and classified as either normal, carrier or having mild, moderate or advanced POAG. A 0.1‐mL sample of AH was drawn from the anterior chamber of each dog in the study and frozen on dry ice. A modified Coomassie stain and Western blot, using a polyclonal rabbit antihuman myocilin antibody (Santa Cruz Biotechnologies, Santa Cruz, CA), was run on each sample to compare the myocilin levels. A purified human trabecular meshwork excreted myocilin protein sample was used as a control (Alcon Research Laboratories, Fort Worth, TX) and its band/densitometry measurement was defined as one unit of myocilin for comparisons. Results Comparisons of AH myocilin levels differed among normal laboratory quality, genetic carrier, and POAG Beagles at different stages of the disease. In the normal laboratory, Beagles the AH myocilin measured 0.817 ± 0.075 units (mean ± SEM); in the carrier Beagles the AH myocilin was 3.117 ± 0.290 units. As POAG progressed, myocilin protein levels also increased to 6.097 ± 0.810, 8.844 ± 1.079, and 17.228 ± 1.198 units in the early, moderate, and advanced forms, respectively. Overall comparisons between normal, carrier and all POAG Beagles combined showed significant differences (P < 0.0010). Individual comparisons between normal and carrier eyes showed significant differences (P < 0.0193). Comparisons between normal and all POAG eyes also showed significant differences (P < 0.0426). Conclusion This study shows myocilin protein is present in normal Beagles, markedly increased in POAG Beagles, and mildly increased in genetic carrier Beagles. There is a strong correlation between amounts of AH myocilin protein and the presence and severity of POAG. The exact role of AH myocilin levels in the genesis of ocular hypertension remains unresolved, but myocilin may adversely affect AH outflow.     

Ascorbic acid ameliorates dysregulated folliculogenesis induced by mono-(2-ethylhexyl)phthalate in neonatal mouse ovaries via reducing ovarian oxidative stress

Phthalates, including di-(2-ethylhexyl)phthalate (DEHP), are common industrial chemicals in the environment. Recent evidence indicates that DEHP and its active metabolite mono-(2-ethylhexyl)phthalate (MEHP) negatively modulate reproductive functions and induce reactive oxygen species. Ascorbic acid (AA) is a dietary requirement for primates, and it acts as a potent free radical scavenger to protect tissues against oxidative stress. In this study, to investigate the toxic effects of MEHP on the follicle development and the beneficial role of AA, neonatal mouse ovaries were treated with different concentrations of MEHP with or without AA for 6 days. Then, the follicle constitution and oxidative status were compared in different groups. Results showed MEHP accelerated primordial follicle recruitment by increasing the percentage of primary and secondary follicles and decreasing the percentage of primordial follicles in the ovaries. Moreover, MEHP-induced ovarian oxidative stress by significantly increasing malondialdehyde (MDA) concentration and the expression of GSS and SOD1. When ovaries were co-administrated with MEHP and AA, follicle constitution was normalized, and the oxidative status was significantly decreased. These results suggested that AA ameliorated MEHP-induced ovarian oxidative stress and follicular dysregulation, which attested the clinical significance of AA for ovary protection in the case of MEHP exposure.     

Enhanced Erythrocytic Lipid Peroxides and Reduced Plasma Ascorbic Acid,and Alteration in Blood Trace Elements Level in Dairy Cows with Mastitis

Oxidative stress has been associated in several inflammatory conditions and incriminated in the pathogenesis of many diseases. However, little information is available on the status of plasma antioxidant levels, essential components of important antioxidant enzymes such as copper, zinc and selenium in blood, and the end product of oxidative damage to the erythrocytic polyunsaturated fatty acids in inflammatory udder conditions. Blood samples were collected from three groups of dairy cows, with 21 in each group: animals with healthy udder, clinical mastitis, and subclinical mastitis. These animals were randomly selected from a herd on the basis of the California mastitis test, somatic cell count and total bacterial count. The mean plasma ascorbic acid concentration was significantly lower in cows with subclinical (p = 0.004) and clinical mastitis (p = 0.000) and the erythrocytic lipid peroxide levels were significantly (p = 0.000) higher in clinical mastitis as compared to controls. There was a significant decrease in mean blood zinc concentration in subclinical (p = 0.005) and clinical mastitis (p = 0.000), but an increase in mean blood copper level in the clinical mastitis group. It was concluded that the blood antioxidant status declines in inflammatory udder conditions, suggesting that incorporation of antioxidants may help in better management of mastitis in dairy cows.     

Population pharmacokinetics of marbofloxacin in aqueous humor after intravenous administration in dogs

OBJECTIVE: To evaluate, by use of population pharmacokinetics, the disposition of marbofloxacin in the aqueous humor after IV administration in dogs and identify its potential usefulness in the prophylaxis and treatment of intraocular infection. ANIMALS: 63 dogs. METHODS: Dogs received a single dose of marbofloxacin (2 mg x kg(-1), IV) at various time intervals before cataract surgery. Aqueous humor and blood samples were collected at the beginning of surgery. Marbofloxacin concentrations were measured by high-pressure liquid chromatography. Data were analyzed with a nonlinear mixed-effect model and, by use of population pharmacokinetic parameters, the time course of aqueous humor concentration was simulated for single doses of 3, 4, and 5.5 mg x kg(-1) IV. Pharmacodynamic surrogate markers and measured aqueous humor concentrations were used to predict in vivo antimicrobial activity. RESULTS: A maximum marbofloxacin concentration of 0.41 +/- 0.17 microg x mL(-1) was reached in the aqueous humor 3.5 hours after IV administration. In the post-distributive phase, marbofloxacin disappeared from aqueous humor with a half-life of 780 minutes. The percentage penetration into the aqueous humor was 38%. Predictors of antimicrobial effects of marbofloxacin (2 mg x kg(-1), IV) indicated that growth of the enterobacteriaceae and certain staphylococcal species would be inhibited in the aqueous humor. Marbofloxacin administered IV at a dose of 5.5 mg x kg(-1) would be predicted to inhibit growth of Pseudomonas aeruginosa and all strains of staphylococci but would not eradicate streptococcal infections. CONCLUSIONS AND CLINICAL RELEVANCE: Marbofloxacin administered IV can penetrate the aqueous humor of canine eyes and may be suitable for prophylaxis or treatment of certain anterior chamber infections.     

Aqueous humor and plasma concentrations of a compounded 0.2% solution of terbinafine following topical ocular administration to normal equine eyes

Objective To determine the transcorneal penetration and systemic absorption of a compounded 0.2% terbinafine solution following repeated topical administration to normal equine eyes. Sample population Six healthy adult horses with normal ocular examinations. Procedures One eye of each horse received 0.2 mL of a compounded 0.2% terbinafine solution every 4 h for seven doses. During the 1 h following administration of the final dose, multiple peripheral blood samples were obtained, and a single aqueous humor (AH) sample was collected at the end of the hour. AH and plasma concentrations of terbinafine were determined using high pressure liquid chromatography (HPLC). Stability of the formulation was assessed with HPLC analysis over a 14‐day time period. Results Terbinafine was not detected in the AH or plasma of any horse at any time point. No signs of ocular irritation or systemic toxicity were noted in any horse at any time point. The solution was stable over 14 days. Conclusion Topical ocular administration of compounded 0.2% terbinafine solution does not result in detectable AH or plasma levels following administration to normal equine eyes, suggesting its use for deep corneal or intraocular fungal infections in equine ophthalmology may be limited.     

外源抗坏血酸对香蕉常温后熟期果实品质的影响

为探究外源抗坏血酸对香蕉果实常温后熟期品质变化的影响,本研究以 ‘巴西’香蕉为材料,用0.5% (质量体积百分比W/V)的抗坏血酸浸泡处理,以清水浸泡为对照,0.03 mm厚聚乙烯打孔薄膜袋包装后,于常温(25±2)℃、相对湿度85%-95%的环境中贮藏。研究结果表明抗坏血酸处理有效抑制香蕉果实常温后熟期呼吸和乙烯释放速率,保持果实的硬度和色度,抑制可溶性固形物和维生素C含量的减少,同时有效抑制电导率的上升和丙二醛的积累。可见,抗坏血酸能够有效延缓香蕉的衰老进程,保持香蕉果实品质,抑制采后香蕉果实膜脂过氧化的发生,从而延长香蕉果实贮藏期。     

Effect of three treatment protocols on acute ocular hypertension after phacoemulsification and aspiration of cataracts in dogs

Objective  To compare the effect of topical latanoprost, intracameral carbachol, or no adjunctive medical therapy on the development of acute postoperative hypertension (POH) and inflammation after routine phacoemulsification and aspiration (PA) of cataracts in dogs.
Design  Retrospective study.
Procedures  Dogs received either one drop of topical 0.005% latanoprost (21 dogs, 39 eyes), an intracameral injection of 0.3 mL of 0.01% carbachol (15 dogs, 30 eyes), or no adjunctive therapy (46 dogs, 90 eyes) immediately following PA of cataract(s). Intraocular pressure (IOP) was measured in all dogs 2 and 4 h after surgery. IOP was measured and aqueous flare assessed at 8 am the day after surgery.
Results  Carbachol-treated dogs had significantly higher mean IOP (33.2 ± SD 20.8 mmHg) 2 h after surgery than dogs receiving no adjunctive therapy (22.0 ± SD 14.1 mmHg) ( P  =  0 .049). There were no significant differences in IOP among groups at any other time point. There were no significant differences in number of POH episodes between dogs treated with carbachol (47%), latanoprost (29%), or dogs that received no adjunctive therapy (33%). There were no significant differences in mean aqueous flare grade between eyes treated with latanoprost (1.7 ± SD 0.4) or carbachol (1.4 ± SD 0.6), and eyes that received no adjunctive therapy (1.7 ± SD 0.4).
Conclusions  Topical 0.005% latanoprost or intracameral injection of 0.3 mL of 0.01% carbachol after PA in dogs did not reduce POH or increase intraocular inflammation compared to dogs not receiving adjunctive therapy after PA of cataracts.     

Residual effect after salmon oil supplementation on semen quality and serum levels of testosterone in dogs

The objective of present study was to evaluate the effects of oral supplementation of salmon oil on seminal parameters and testosterone serum levels in dogs, following also the residual effects for 60 days after treatment. Nine healthy male dogs with proven fertility, weighing between 10 and 36 kg, ageing from 2 to 11 years, of different breeds, fed diets supplemented with salmon oil at the manufacturer's recommended dosage. The parameters measured were sperm volume, motility, vigour, normal morphology and concentration, live/dead ratio, membrane viability by means of HOST test and serum testosterone levels. Evaluations occurred at baseline (D0), after 90 days of supplementation (D90) and at the end of the experiment, 60 days after supplementation cessation (D150). Results (mean ± SD) obtained at time D0, D90 and D150 were as follows: motility of 76.66% ± 13.7, 92.77 ± 4.41 and 93.0 ± 7.90 (p = .001); normal spermatozoa of 69.11% ± 24.90, 90.00% ± 5.15 and 80.66 ± 16.04 (p = .05); live/dead (%) from 64.44 ± 22.86 to 85.33 ± 8.41 (p = .001); and spermatozoa (%) with integral membrane in the membrane integrity (HOST) test ranging from 76.44 ± 20.74 to 91.22 ± 4.68 (p = .05). Serum levels of testosterone (ng/ml) increased from 5.50 ± 1.13 to 8.84 ± 1.13 at D90 (p = .003) and decreased after 2 months (D150) to 5.13 ± 1.13. In conclusion, a 90‐day supplementation with salmon oil had a positive influence on semen quality and serum testosterone levels. The supplementation of omegas 3 and 6 at the ratio of 10:1 for 90 days determined an increase in concentration and motility of the sperm, and these effects were maintained for 60 days, with the only exception of testosterone levels.     

植物体内自由基清除剂抗坏血酸研究进展

抗坏血酸(Ascorbic Acid,AsA)作为植物体内一种重要的自由基清除剂,在减少膜脂过氧化作用对细胞造成的损伤、增强植物抵抗逆境胁迫能力等过程中起着非常重要的作用。通过分析比较AsA的生物合成途径,总结AsA参与细胞分裂与伸长、作为辅酶参与细胞壁的形成、光保护作用、作为电子传递与信号转导的载体等生理功能,系统阐述AsA在植物逆境胁迫（主要包括温度胁迫、盐胁迫、生物胁迫等）中的保护机制及功能,强调AsA研究对于了解自由基清除剂在植物抗逆性作用以及清除自由基机理方面具有重要作用。本研究提出在牧草种子劣变方面AsA研究的不足,并分析展望了AsA研究发展的趋势。     

Ciprofloxacin and ofloxacin aqueous humor concentrations after topical administration in dogs undergoing cataract surgery

OBJECTIVE: To determine if preoperative topical administration of ciprofloxacin or ofloxacin results in aqueous humor drug concentrations that exceed the MIC(90) of common ocular contaminants in dogs undergoing cataract surgery. PROCEDURES: Twelve dogs were treated with topical 0.3% ciprofloxacin and 13 dogs with topical 0.3% ofloxacin once the night before surgery, and then with 1 drop of ciprofloxacin or ofloxacin every 15 min for 2 h immediately before surgery. Aqueous humor samples were taken from each eye immediately before the incision was made and frozen at -70 degrees C. First eye samples (S1) were taken closer to the time of topical treatments than second eye samples (S2). Samples were analyzed by high performance liquid chromatography (HPLC) at the North Carolina State University (NCSU) Clinical Pharmacology laboratory. RESULTS: In ciprofloxacin patients, S1 concentrations were 0.03-0.69 (0.17 median) microg/mL, and S2 concentrations were 0.09-0.95 (0.36 median) microg/mL. Aqueous humor concentrations did not exceed the MIC90 of Streptococcus sp. Few eyes (1/12 OU) exceeded the MIC90 for Staphylococcus sp. or Corynebacterium sp.; moderate numbers (5/12 S1; 8/12 S2) exceeded the MIC90 for E. coli. In ofloxacin patients, S1 concentrations were 0.48-2.81 (1.05 median) microg/mL, and S2 concentrations were 0.45-3.63 (1.30 median) microg/mL. Although few eyes (相似文献   

温度、柠檬酸和抗环血酸对冬蜜中多酚氧化酶活性的影响

以儿茶酚为底物,采用紫外分光光度法,测定冬蜜中多酚氧化酶(PPO)的活性。结果表明,在20～90℃的条件下,冬蜜中PPO的活性,随着温度的升高而增强;在冬蜜中添加0.1%柠檬酸时,对其PPO活性抑制效果最好;但添加0.1%抗坏血酸(Vc)时,对其PPO活性抑制作用较弱。     

抗坏血酸和水杨酸丸衣对NaCl胁迫下紫花苜蓿种子发芽特性的影响

为探究抗坏血酸（AsA）和水杨酸（SA）丸衣对NaCl胁迫下紫花苜蓿种子萌发的缓解效应,以紫花苜蓿种子为试验材料,采用不同浓度的AsA（1、2、4、8 mmol·L^-1）和SA（1、5、10、20 mmol·L^-1）对紫花苜蓿种子拌种后进行丸衣处理（未丸衣种子为CK₁,不做拌种处理的丸衣种子记为CK₂）,分析丸衣种子在NaCl胁迫下发芽特性的变化,以期获得能有效缓解紫花苜蓿种子盐胁迫效应的种子丸衣配方。结果表明,1.25%和1.50%的NaCl胁迫显著（P<0.05）降低了紫花苜蓿未丸衣种子的发芽率、发芽势、种苗长和种苗重,延长了平均发芽时间。AsA在1.25%NaCl胁迫下缓解效果不明显,4 mmol·L^-1AsA拌种后丸衣处理,能提高1.50%NaCl胁迫下紫花苜蓿种子的发芽率、种苗长。1 mmol·L^-1SA拌种后丸衣处理,能提高1.50%NaCl胁迫下紫花苜蓿种子的发芽势、发芽率和种苗长,而20 mmol·L^-1SA拌种后丸衣处理降低了紫花苜蓿种子的发芽率。说明AsA和SA丸衣对NaCl胁迫的缓解作用与其拌种浓度和NaCl胁迫浓度有关。本试验筛选出4 mmol·L^-1AsA和1 mmol·L^-1SA拌种丸衣配方对NaCl胁迫的缓解作用最优,该结果为AsA和SA丸衣应用于紫花苜蓿种子缓解NaCl胁迫提供依据。