Impact of cerebral hypoperfusion-reperfusion on optic nerve integrity and visual function in the DBA/2J mouse model of glaucoma.

OBJECTIVE: One of the most important risk factors for developing a glaucomatous optic neuropathy is elevated intraocular pressure. Moreover, mechanisms such as altered perfusion have been postulated to injure the optical path. In a mouse model, we compare first negative effects of cerebral perfusion/reperfusion on the optic nerve structure versus alterations by elevated intraocular pressure. Second, we compare the alterations by isolated hypoperfusion-reperfusion and isolated intraocular pressure to the combination of both. METHODS AND ANALYSIS: Mice were divided in four groups: (1) controls; (2) perfusion altered mice that underwent transient bi-common carotid artery occlusion (BCCAO) for 40 min; (3) glaucoma group (DBA/2J mice); (4) combined glaucoma and altered perfusion (DBA/2J mice with transient BCCAO). Optic nerve sections were stereologically examined 10-12 weeks after intervention. RESULTS: All experimental groups showed a decreased total axon number per optic nerve compared with controls. In DBA/2J and combined DBA/2J & BCCAO mice the significant decrease was roughly 50%, while BCCAO leaded to a 23% reduction of axon number, however reaching significance only in the direct t-test. The difference in axon number between BCCAO and both DBA/2J mice was almost 30%, lacking statistical significance due to a remarkably high variation in both DBA/2J groups. CONCLUSION: Elevated intraocular pressure in the DBA/2J mouse model of glaucoma leads to a much more pronounced optic nerve atrophy compared with transient forebrain hypoperfusion and reperfusion by BCCAO. A supposed worsening effect of an altered perfusion added to the pressure-related damage could not be detected.

Diurnal and menstrual cycles in body temperature are regulated differently: a 28-day ambulatory study in healthy women with thermal discomfort of cold extremities and controls.

Diurnal cycle variations in body-heat loss and heat production, and their resulting core body temperature (CBT), are relatively well investigated; however, little is known about their variations across the menstrual cycle under ambulatory conditions. The main purpose of this study was to determine whether menstrual cycle variations in distal and proximal skin temperatures exhibit similar patterns to those of diurnal variations, with lower internal heat conductance when CBT is high, i.e. during the luteal phase. Furthermore, we tested these relationships in two groups of women, with and without thermal discomfort of cold extremities (TDCE). In total, 19 healthy eumenorrheic women with regular menstrual cycles (28-32 days), 9 with habitual TDCE (ages 29 ± 1.5 year; BMI 20.1 ± 0.4) and 10 controls without these symptoms (CON: aged 27 ± 0.8 year; BMI 22.7 ± 0.6; p < 0.004 different to TDCE) took part in the study. Twenty-eight days continuous ambulatory skin temperature measurements of distal (mean of hands and feet) and proximal (mean of sternum and infraclavicular regions) skin regions, thighs, and calves were carried out under real-life, ambulatory conditions (i-Buttons® skin probes, sampling rate: 2.5 min). The distal minus proximal skin temperature gradient (DPG) provided a valuable measure for heat redistribution from the core to the shell, and, hence, for internal heat conduction. Additionally, basal body temperature was measured sublingually directly after waking up in bed. Mean diurnal amplitudes in skin temperatures increased from proximal to distal skin regions and the 24-h mean values were inversely related. TDCE compared to CON showed significantly lower hand skin temperatures and DPG during daytime. However, menstrual cycle phase did not modify these diurnal patterns, indicating that menstrual and diurnal cycle variations in skin temperatures reveal additive effects. Most striking was the finding that all measured skin temperatures, together with basal body temperature, revealed a similar menstrual cycle variation (independent of BMI), with highest and lowest values during the luteal and follicular phases, respectively. These findings lead to the conclusion that in contrast to diurnal cycle, variations in CBT variation across the menstrual cycle cannot be explained by changes in internal heat conduction under ambulatory conditions. Although no measurements of metabolic heat production were carried out increased metabolic heat generation during the luteal phase seems to be the most plausible explanation for similar body temperature increases.

Differences in gene expression in lymphocytes of patients with high-tension, PEX, and normal-tension glaucoma and in healthy subjects.

PURPOSE: Purpose Differences in the gene expression of leukocytes between patients with normal-tension glaucoma (NTG) and controls have been described. This study was performed in order to detect the differences in gene expression in peripheral lymphocytes in patients with primary open-angle glaucoma (POAG), patients with pseudoexfoliation glaucoma (PEX), and patients with NTG, and in healthy subjects. METHODS: Ten patients with POAG, 11 patients with PEX, 10 patients with NTG, and 42 sex- and age-matched healthy persons were recruited. All study subjects were Caucasian. Twenty-two preselected genes were chosen and their expression in blood lymphocytes was quantified by real-time PCR. First, a univariate comparison among all groups was performed using the nonparametric Friedman test. Second, an L1 penalized logistic regression was performed. RESULTS: Using the Friedman test to compare the 4 groups, 9 genes showed a different expression (p<0.05). Comparing the controls vs patients with POAG, 8 genes were differently expressed (p<0.05). Comparing patients with PEX vs controls, 9 genes were significantly different (p≤0.05). The statistical analysis of patients with NTG vs controls showed a difference in gene expression of 7 genes (p≤0.05). All these genes were upregulated in the glaucoma groups compared with the controls. The genes RhoGDI and RAR showed the most significant statistical difference in the L1-penalized logistic regression. The genes overexpressed in POAG/PEX differed from the ones in NTG. CONCLUSIONS: In this masked study among the preselected 22 genes, several genes are overexpressed in the blood lymphocytes of Caucasian patients with glaucoma compared with the controls. The genes upregulated in POAG/PEX differed from the ones in NTG.

Object recognition memory and BDNF expression are reduced in young TgCRND8 mice.

The TgCRND8 mouse model of Alzheimer's disease exhibits progressive cortical and hippocampal ß-amyloid accumulation, resulting in plaque pathology and spatial memory impairment by 3 months of age. We tested whether TgCRND8 cognitive function is disrupted prior to the appearance of macroscopic plaques in an object recognition task. We found profound deficits in 8-week-old mice. Animals this age were not impaired on the Morris water maze task. TgCRND8 and littermate controls did not differ in their duration of object exploration or optokinetic responses. Thus, visual and motor dysfunction did not confound the phenotype. Object memory deficits point to the frontal cortex and hippocampus as early targets of functional disruption. Indeed, we observed altered levels of brain-derived neurotrophic factor (BDNF) messenger ribonucleic acid (mRNA) in these brain regions of preplaque TgCRND8 mice. Our findings suggest that object recognition provides an early index of cognitive impairment associated with amyloid exposure and reduced brain-derived neurotrophic factor expression in the TgCRND8 mouse.

Plasma citrate levels as a potential biomarker for glaucoma.

PURPOSE: To determine the possibility of plasma citrate as a biomarker in patients with glaucoma. METHODS: Twenty-one consecutive Caucasian patients with glaucoma and 21 sex- and age-matched controls were investigated. Plasma citrate, plasma creatinine, urine citrate, and urine creatinine were analyzed by ion chromatography. Mean (±standard deviation) concentrations and the calculated fractional citrate excretions were compared using the Mann-Whitney test. Sensitivity and specificity to detect glaucoma using plasma citrate levels were calculated. RESULTS: The mean plasma citrate (104.8±23.2 vs. 128.2±31.1 µmol/L; P=0.01) concentrations were significantly lower among the patients with glaucoma, whereas the mean urine citrate concentrations (1.7±0.9 vs. 2.8±1.9 µmol/L; P=0.07) were slightly lower. Mean plasma and mean urine creatinine concentrations showed no significant differences (plasma creatinine: 63.0±16.7 vs. 63.4±15.5 µmol/L; P=0.72; urine creatinine: 9.6±5.1 vs. 11.5±8.4 µmol/L; P=0.67). The calculated fractional citrate excretions were also not different with 12.1% versus 13.6% (P=0.37). Setting the cut-off limit at 110 µmol/L, the plasma citrate level evaluation would have a sensitivity of 66.7% and a specificity of 71.4% to detect glaucoma. CONCLUSION: In this masked study, plasma citrate levels were significantly decreased in Caucasian patients with glaucoma giving the possibility to use them eventually as a biomarker. The kidney function was normal in both groups, leaving the etiology of this hypocitraemia yet unexplained.

Circumferential viscocanalostomy and suture canal distension (canaloplasty) for whites with open-angle glaucoma.

PURPOSE: To assess the safety and efficacy of canaloplasty (360-degree viscodilation and tensioning of the Schlemm canal) in Whites with open-angle glaucoma (OAG). METHODS: In a prospective study, 32 consecutive patients with medically uncontrolled OAG underwent primary canaloplasty with a follow-up time of more than 1 year. Laser goniopuncture was performed if postoperative intraocular pressure (IOP) was above 16 mmHg. IOP, number of antiglaucomatous medications, best-corrected visual acuity, and intraoperative and postoperative complications were recorded. Complete success was defined as an IOP ≤21, 18, and 16 mm Hg without medications, and qualified success with or without medications, respectively. RESULTS: The mean IOP dropped from 27.3±5.6 mm Hg preoperatively to 12.8±1.5 mm Hg at 12 months and 13.1±1.2 mm Hg at 18 months (P<0.001). The complete success rate of an IOP ≤21, 18, and 16 mm Hg was 93.8% [95% confidence interval (CI) 0.86-1.0], 84.4% (95% CI 0.73-0.98), and 74.9% (95% CI 0.61-0.92), respectively, at 12 months. Laser goniopuncture was performed on 6 eyes (18.1%) 3.3±2.1 months postoperatively. The mean IOP was 20.6±4.2 mm Hg before and 14.2±2.2 mm Hg after goniopuncture. The number of medications dropped from 2.7±0.5 before surgery to 0.1±0.3 after surgery (P<0.001). The postoperative best-corrected visual acuity at last visit (0.38±0.45; range: 0 to 1.8) was comparable with that of preoperative values (0.36±SD 0.37; range: 0 to 1.6) (P=0.42). In all but 1 eye, canaloplasty was completed. Minor intraoperative or postoperative complications like Descemet membrane detachment in 2 eyes, elevated IOP in 1 eye, and suprachoroidal passage of the catheter in 4 eyes were encountered. In 1 eye, circumferential cannulation of the Schlemm canal was impossible. CONCLUSIONS: Canaloplasty seems to be a promising and effective surgical procedure in Whites with OAG. Postoperative IOP levels are in the low-to-mid-teens. The procedure can be regarded as safe, but has its own profile of complications.

Retinal vein occlusions: The potential impact of a dysregulation of the retinal veins.

A retinal vein occlusion (RVO) is a sight threatening disease. It can be divided into central vein occlusion and branch retinal vein occlusion. The pathogenesis of the condition remains to be solved. Mechanical compression of the vessel wall or thrombotic occlusion of the vessel lumen, sometimes combined with rheological disorders, are often assumed pathomechanisms. Accordingly, the therapy relies either on mechanical decompression, lyses of thrombi or improvement of rheology. A number of observations however, such as the relationship of RVO to atherosclerotic risk factors, spontaneous reversibility particularly in young patients, rest flow observed in angiography, occlusion despite anticoagulation or thrombocytopenia and finally the positive effect of anti-VEGF therapy are not explained by the present pathogenetic concept. As a new concept we propose a local venous constriction induced by vasoconstrictive molecules diffusing from neighbouring diseased arteries and/or from other neighbouring (hypoxic) tissues. Recognizing these postulated conditions might lead to an earlier identification of impending vein occlusions as well as to a treatment more tailored to the risk factor constellation of the particular patient.

Identification of lymphatics in the ciliary body of the human eye: a novel "uveolymphatic" outflow pathway.

Impaired aqueous humor flow from the eye may lead to elevated intraocular pressure and glaucoma. Drainage of aqueous fluid from the eye occurs through established routes that include conventional outflow via the trabecular meshwork, and an unconventional or uveoscleral outflow pathway involving the ciliary body. Based on the assumption that the eye lacks a lymphatic circulation, the possible role of lymphatics in the less well defined uveoscleral pathway has been largely ignored. Advances in lymphatic research have identified specific lymphatic markers such as podoplanin, a transmembrane mucin-type glycoprotein, and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1). Lymphatic channels were identified in the human ciliary body using immunofluorescence with D2-40 antibody for podoplanin, and LYVE-1 antibody. In keeping with the criteria for lymphatic vessels in conjunctiva used as positive control, D2-40 and LYVE-1-positive lymphatic channels in the ciliary body had a distinct lumen, were negative for blood vessel endothelial cell marker CD34, and were surrounded by either discontinuous or no collagen IV-positive basement membrane. Cryo-immunogold electron microscopy confirmed the presence D2-40-immunoreactivity in lymphatic endothelium in the human ciliary body. Fluorescent nanospheres injected into the anterior chamber of the sheep eye were detected in LYVE-1-positive channels of the ciliary body 15, 30, and 45 min following injection. Four hours following intracameral injection, Iodine-125 radio-labeled human serum albumin injected into the sheep eye (n = 5) was drained preferentially into cervical, retropharyngeal, submandibular and preauricular lymph nodes in the head and neck region compared to reference popliteal lymph nodes (P < 0.05). These findings collectively indicate the presence of distinct lymphatic channels in the human ciliary body, and that fluid and solutes flow at least partially through this system. The discovery of a uveolymphatic pathway in the eye is novel and highly relevant to studies of glaucoma and other eye diseases.