The effect and safety of intravitreal injection of ranibizumab and bevacizumab on the corneal endothelium in the treatment of diabetic macular edema

Objective: To investigate the effect and safety of intravitreal injection (IVI) of bevacizumab and ranibizumab on corneal endothelial cell count and morphology in patients with diabetic macular edema.

Materials and methods: A total of 60 eyes from 60 consecutive patients who received 0.5?mg/0.05?ml IVIs of bevacizumab (n?=?30, IVB group) or 1.25?mg/0.05?ml ranibizumab (n?=?30, IVR group) for three consecutive months were investigated prospectively. Specular microscopy was performed to evaluate endothelial cell count, the percentage of hexagonal cells (pleomorphism), and the coefficient of variation of the cell size (polymegathism); optical biometry was performed to evaluate central corneal thickness. Results before injection and 1 month after the first and third injections were compared.

Results: The groups were matched for age (p?=?0.11) and gender (p?=?0.32). There was no significant difference in endothelial cell count (IVB group, p?=?0.66; IVR group, p?=?0.74), pleomorphism (IVB group, p?=?0.44; IVR group, p?=?0.88) and polymegathism (IVB group, p?=?0.21; IVR group, p?=?0.24) before injection or 1 month after the first and third injections. There was also no difference in central corneal thickness (IVB group, p?=?0.15; IVR group, p?=?0.58) before injection or 1 month after the first and third injections.

Conclusion: Monthly 1.25?mg/0.05?ml IVIs of bevacizumab or 0.5?mg/0.05?ml of ranibizumab for three consecutive months in the treatment of diabetic macular edema does not affect corneal morphology and has no harmful effects on the endothelium.     

Impact of intravitreal triamcinolone acetonide versus intravitreal bevacizumab on retrobulbar hemodynamic in patients with diabetic macular edema

Purpose: To evaluate and compare retrobulbar hemodynamic changes measured with color Doppler imaging (CDI) in diabetic patients receiving intravitreal triamcinolone acetonide (IVTA) versus bevacizumab.

Methods: Patients with diffuse diabetic macular edema were assessed prospectively by CDI following intravitreal injection of triamcinolone acetonide (group I, 12 eyes) versus bevacizumab (group II, 14 eyes). CDI was used to measure the peak systolic velocity (PSV), end diastolic velocity (EDV) and the resistive index (RI) of the central retinal artery (CRA), ophthalmic artery (OA) and posterior ciliary arteries (PCA) one day preoperatively and one week postoperatively.

Results: In group I, EDV of OA and CRA decreased significantly (p?=?0.007 and 0.018, respectively). The PSV and RI of PCA decreased significantly (p?=?0.035 and 0.002, respectively). In group II, both the PSV and EDV of the CRA decreased significantly (p?=?0.000). Comparing the percentage of change in both groups, PSV of the CRA decreased significantly in group II (p?=?0.034), while IVTA has more significant effect on the ophthalmic artery hemodynamic parameters as EDV decreased and RI increased significantly (p?=?0.045 and 0.043, respectively)

Conclusion: Intravitreal injections of triamcinolone acetonide and bevacizumab have a significant effect on the ocular hemodynamic. The effect of bevacizumab is statistically significant on the PSV of CRA compared to IVTA.     

Protective effects of N-acetylcysteine on triamcinolone acetonide-induced lens damage in rats

Objective: To examine the relationship of cataract forming effect of intravitreal triamcinolone acetonide (IVTA) injection with oxidative status and the effect of N-acetylcysteine (NAC) on these alterations.

Materials and methods: Twenty-six Wistar-Albino rats were included in the study. Rats were assigned into four groups as follows: intravitreal saline injection group (controls); IVTA injection group; IVTA?+?intraperitoneal NAC injection group (IVTA?+?NAC); and intraperitoneal NAC injection group (NAC). Triamcinolone acetonide was intravitreally injected at a dose of 1?mg. NAC was intraperitoneally injected at a dose of 150?µg/g body weight. Animals were sacrificed and lens specimens were analyzed for levels of malondialdehyde (MDA) and protein carbonyl (PC) and activities of glutathione (GSH) and glutathione peroxidase (GSH-Px).

Results: We found that the MDA and PC levels of lenses were increased in the IVTA group (p?p?p?Conclusion: These results indicate that the NAC produces a protective mechanism against IVTA-induced cataract and suggest a role of oxidative stress in pathogenesis.     

Ocular surface effects of repeated application of povidone iodine in patients receiving frequent intravitreal injections

Background: The aim of this study was to investigate the patient reported symptoms and objective signs of tear film and ocular surface abnormalities experienced by patients undergoing repeated exposure to povidone iodine as a consequence of requiring frequent intravitreal injections for wet macular degeneration.

Methods: This was a prospective study of consecutive patients who had received recent povidone 5% solution for sterile preparation of intravitreal injection less than 3?months prior to inclusion with a total of at least 3 intravitreal injections for macular degeneration. Each patient had one study eye which was undergoing regular intravitreal injection and a fellow eye which was not undergoing any injections. Each patient underwent evaluations of various tear film parameters on a single occasion for both eyes. The primary outcome was severity of dry eye symptoms as measured by the Schein dry eye questionnaire. The secondary outcomes were tear film osmolarity and corneal punctate staining using the Oxford Grading Scale.

Results: A total of 90 patients were included in the study. 43.3% n?=?39, were using ocular lubricating medication on a regular basis. A significantly greater proportion of study eyes had a Schein dry eye questionnaire score of 7 or higher; 12.2%, n?=?11 amongst study eyes vs 4.4%, n?=?4 amongst control, fellow eyes (p?p?=?0.087). The study eyes had statistically significantly worse corneal staining as determined by the Oxford grading scale; 0.69 in study eyes vs 0.58 in control, fellow eyes (p?=?0.02).

Conclusion: Our results confirm the detrimental impact of repeated application of povidone iodine for intravitreal injection procedures on symptoms of dry eyes as experienced and reported by patients.     

Effect of intravitreal injection of dexamethasone implant on corneal endothelium in macular edema due to retinal vein occlusion

Objective: To evaluate the effects of dexamethasone (DEX) implant (Ozurdex®) on corneal endothelium in patients with retinal vein occlusion complicated with macular edema.

Materials and methods: Patients (n?=?31) received 1–3 intravitreal DEX implants in one eye. Measurements were intraocular pressure (IOP) at baseline and 1, 3, and 6 months after the first intravitreal injection and corneal specular microscopy and central corneal thickness (CCT) at baseline and 1 and 6 months. We analyzed endothelial cell density (ECD), coefficient of variation of cell size (CV), and percentage of hexagonality.

Results: Mean follow-up period was 9.7?±?3.3 months. Mean number of injections was 1.5?±?0.8. Mean IOP values were 15.6?±?2.6?mm Hg at baseline, 17.7?±?3.6?mm Hg at one month, 16.4?±?4.1?mm Hg at three months, and 16.0?±?2.7?mm Hg at six months. There was a significant difference in mean IOPs at one month and six months (p?=?0.008). There were no significant differences in mean ECD (p?=?0.375), CV (p?=?0.661), percentage of hexagonality (p?=?0.287), and CCT (p?=?0.331).

Conclusion: Although intravitreal injection of 0.7?mg DEX causes moderate elevation of IOP, it does not seem to have detrimental effects on corneal endothelium at six months.     

Morphologic changes and visual outcomes in resolved central serous chorioretinopathy treated with ranibizumab

Context: Central serous chorioretinopathy (CSC).

Objective: To evaluate the effect of intravitreal ranibizumab injection and the correlation between foveal morphologic changes and visual outcomes in patients with resolved CSC.

Materials and methods: We measured outer nuclear layer (ONL) thickness, outer layer (OL) thickness and evaluated the integrity of the photoreceptor inner-outer segment (IS/OS) junction, the status of the external limiting membrane (ELM) at the central fovea using spectral-domain optical coherence tomography (OCT) in 35 eyes of 35 patients with resolved CSC. The eyes were divided into two groups: The initial medical treatment administered to Group1 (n?=?17) then received intravitreal ranibizumab injections, Group 2 (n?=?18) received medical treatment. Group 3 was composed of normal eyes (n?=?20, as a control). We also investigated a correlation between the ONL thickness and best corrected visual acuity (BCVA).

Results: The mean age was 45.7?±?7.2 (ranged from 27 to 55 years). The mean follow-up period was 14.2 months (minimum 6, maximum 24 months). The mean ONL and OL thickness in Group 1 were significantly thinner than Group 3 (p?r?=?0.681, p?=?0.001). Thirty-tree patients had improvement in BCVA after treatment. Discontinuity of the IS/OS junction was found in 15 eyes (88.2%) in Group 1, in 5 eyes (27.7%) in Group 2 and in no eyes in Group 3.

Discussion: We demonstrated that prolonged serous detachment results in photoreceptor cell loss (apoptosis) and thinning of the ONL. Thinning of the ONL correlates with poorer vision, which has been found by other investigators. Furthermore, vascular endothelial growth factor (VEGF) may be neuroprotective to the photoreceptors which might explain the additional thinning in the patients treated with ranibizumab. This raises the possibility that treatment with VEGF inhibitors may be unfavourable to patients with CSC, even though it speeds recovery and vision does improve.

Conclusion: Intravitreal ranibizumab injection leads to thinning of the ONL and the OL in patients with resolved CSC. The ONL thickness reduction and discontinuity of the IS/OS junction results in poor visual prognosis in resolved CSC eyes.     

Effects of sunitinib and bevacizumab on VEGF and miRNA levels on corneal neovascularization

Aim: To evaluate the effects of sunitinib (0.5?mg/ml) and bevacizumab (5?mg/ml) on VEGF-A, VEGFR-2 and microRNA (miRNA) levels on corneal neovascularization (CNV).

Methods: In this study, CNV was induced by silver nitrate application to the cornea, and 40 Albino male rats were equally divided into four subgroups:

Group 1 (sunitinib): After silver nitrate application to the cornea, 0.5?mg/ml sunitinib eyedrop was administered twice daily for two weeks (n?=?10).

Group 2 (bevacizumab): After silver nitrate application to the cornea, 5?mg/ml bevacizumab eyedrop was administered twice daily for two weeks (n?=?10).

Group 3 (control): After silver nitrate application to the cornea, normal saline eyedrop was administered twice daily for two weeks (n?=?10).

Group 4 (vehicle): After silver nitrate application to the cornea, 1% DMSO eyedrop was administered twice daily for two weeks (n?=?10).

After two weeks from the silver nitrate application, corneas were evaluated by hand-held biomicroscope for their vascularization status. Then, corneas were excised and the expression levels of VEGFR-2, VEGF-A and the common miRNA markers for neovascularization (miR-15?b, miR-16, miR-23a, miR-126, miR-188, miR-210, miR-221, miR-222, miR-410 and miR-423) were evaluated by real-time PCR.

Results: It was seen that the CNV was decreased in sunitinib- and bevacizumab-administered groups compared to the control and DMSO groups. Also, in comparison with the control group; VEGF-A expression was downregulated by nearly 0.75 times in sunitinib group and nearly 0.52 times in bevacizumab group. VEGFR-2 expression was downregulated by 0.89 times in sunitinib group and 0.68 times in bevacizumab group, compared to the control group. miR-15?b, miR-16 and miR-126 levels were statistically lower in sunitinib and bevacizumab groups, but miR-188 and miR-410 levels were two-fold higher compared to the control group. The miR-210 level was found higher only in sunitinib group compared to the control group. There were no statistically significant changes in miR-23a, miR-221, miR-222 and miR-423 levels among the groups.

Conclusion: Topical application of bevacizumab (5?mg/ml) and sunitinib (0.5?mg/ml) decreases the levels of VEGFR-2 and VEGF-A in CNV. Further studies are needed for detailed analysis of genes which are targeted by up- or downregulated miRNAs in this study.     

Does moxifloxacin alter oxidant status in the cornea? An experimental study

Objective: In this experimental study, we investigated the possible effects of intracameral moxifloxacin on oxidative stress parameters and endothelial cell morphology in corneal tissue.

Methods: In total, 30 rats were randomly assigned to three groups of 10 rats: the sham group (Group 1, n?=?10); the control group (Group 2), where balanced salt solution (BSS) was administered at a dose of 0.01?cc (n?=?10); and the treatment group (Group 3), where moxifloxacin was administered at a dose of 0.05?mg/0.01?cc (n?=?10). Total antioxidant status (TAS) and total oxidant status (TOS) in corneal tissue and blood samples were measured and the oxidative stress index (OSI) was calculated. Also, corneal tissue histopathology was evaluated with caspase-3 and caspase-8 staining. Apoptotic activity was also evaluated.

Results: In blood samples, TAS, TOS, and OSI levels were not statistically significantly different (all p?>?0.05). Compared with the sham and control groups, TOS and OSI levels in cornea tissue were significantly different in the moxifloxacin group (all p?p?>?0.05). Compared with the sham and control groups, apoptotic activity was higher in the moxifloxacin group, in both immunohistochemical staining for caspase-3 and caspase-8.

Conclusions: Intracameral moxifloxacin injection seems to be safe systemically, but it may have toxic effects on corneal tissues, as suggested by oxidative stress parameters and a histopathological evaluation.     

Effect of alpha lipoic acid on smoking-induced skin damage

Objective: Exposed to cigarette leads to the formation of reactive oxygen species and the generation of bioactive molecules that can damage skin cells. This investigation was carried out to study possible effects of Alpha Lipoic Acid (ALA) on smoking-induced rat skin injury.

Materials and methods: 28 Spraque–Dawley female rats were allocated into three groups: control group (n?=?8), smoking group (n?=?10; 12 cigarettes/day, 8 weeks) and smoking?+?ALA group (n?=?10; 12 cigarettes/day?+?100?mg/kg, 8 weeks). Experiment group animals were sacrificed under anaesthesia with 10%ketamine?+?2%xylasine at the end of second mounts and then skin examples were taken from the epigastric area. Histochemical (Haematoxylin–Eosin and Masson’s trichrome, immunohistochemical (TNF-α) and biochemical analysis (CAT, MDA and protein carbonylation) were performed on these skin tissues.

Results: Histologically, skin was distinguished normal structure in the control group. In the smoking group, collagen bundles and hair follicle degradation/reduction, sweat gland degeneration, mononuclear cell infiltration in dermis were encountered. In ALA-treated group, all of these changes were improved (p?<?0.05). Collagen bundles structures were appearance more regular than the smoking group . Immunohistologically, intense staining was observed in the smoking group, while very weak staining was observed in control group, weak staining was observed in the ALA-treated group. Biochemically; The CAT activity compared to cigarette group with control was raised high and in ALA group was higher compared to both groups, but not significant (p?>?0.05). MDA; which is indicator of lipid peroxidation was significantly higher in cigarette group than in control group (p?<?0.05) and was significantly lower in ALA group than cigarette (p?<?0.05). Protein carbonylation was higher in cigarette group than the control group but not in the non-significant (p?>?0.05). In the ALA it was significantly lower compared to the control group and cigarette (p?<?0.05).

Conclusions: Based on biochemical and histopathological determinations, the study showed that cigarette smoke can cause degenerative effects on skin tissues in rats. However, ALA has a curative effect on cigarette-induced injuries on the skin tissues by anti-oxidative and anti-inflammatory effects.     

Angiogenesis inhibitors for patients with ovarian cancer: a meta-analysis of 12 randomized controlled trials

Objectives:

To investigate the effects of angiogenesis inhibitors in the treatment of patients with advanced or recurrent ovarian cancer, a meta-analysis was performed and overall survival (OS), progression-free survival (PFS), and toxicity were assessed.

Patients and methods:

The PubMed and Embase databases, and the Cochrane Central Register of Controlled Trials were searched for publications between January 2000 and June 2015. Hazard ratios (HRs) with their 95% confidence intervals (CIs), or data for calculating HRs with 95% CIs were derived.

Results:

The 12 trials in this meta-analysis were divided into three groups: four trials with a VEGF inhibitor (the bevacizumab group), six trials with VEGFR inhibitors (the VEGFRIs group), and two trials with an angiopoietin inhibitor (the trebananib group). PFS improvement was seen in all groups (HR?=?0.61, 95% CI 0.48 to 0.79, P?<?0.001 for bevacizumab; HR?=?0.71, 95% CI 0.59 to 0.87, P?=?0.001 for VEGFRIs; and HR?=?0.67, 95% CI 0.62 to 0.72, P?<?0.001 for trebananib). Regarding OS, bevacizumab showed a trend of improvement (HR?=?0.90, 95% CI 0.80 to 1.01, P?=?0.079), VEGFRIs showed no improvement (HR?=?0.92, 95% CI 0.75 to 1.11, P?=?0.368), and trebananib demonstrated a significant prolongation (HR?=?0.81, 95% CI 0.67 to 0.99, P?=?0.036). Bevacizumab was associated with more class-specific adverse events (RR?=?4.05, 95% CI 1.99 to 8.27, P?<?0.001). Although the toxicity profiles differed, VEGFRIs developed common higher incidences of hypertension, diarrhea, and fatigue. A higher incidence of edema was reported in the trebananib group (RR?=?2.60, 95% CI 0.84 to 8.00, P?=?0.097).

Conclusions:

Anti-angiogenic therapy showed clear PFS benefit with increased toxicity, but its role in OS was undefined for ovarian cancer which emphasized the need for patient selection.     

The prevalence of 4G/5G polymorphism of plasminogen activator inhibitor-1 (PAI-1) gene in central serous chorioretinopathy and its association with plasma PAI-1 levels

Context: Central serous chorioretinopathy (CSCR) is a poorly understood disease and the choroidal circulation abnormality induced by the plasminogen activator inhibitor type 1 (PAI-1) seems to be associated with the pathogenesis. There are many reports indicating that 4G/5G polymorphism of the PAI-1 gene is a risk factor for several diseases related to the elevated serum levels of PAI-1.

Objective: To evaluate the 4G/5G polymorphism of the PAI-1 gene and its association with serum levels of PAI-1 in acute CSCR patients.

Materials and methods: Sixty CSCR patients and 50 healthy control patients were included. The PAI-1 4G/5G was genotyped using the polymerase chain reaction–restriction technique. Serum PAI-1 level was measured using enzyme-linked immunosorbent assay. Demographic data consisting of age, sex, body mass index (BMI) as well as genotype disturbances and serum PAI-1 levels were compared between the groups. Statistical significance for differences in the serum PAI-1 levels of each group with different genotypes was also analyzed.

Results: The CSCR group consisted of 40 male (66.7%) and 20 female (33.3%) patients with a mean age of 46.7?±?8.39 years. The control group consisted of 32 male (64%) and 18 female (36%) healthy subjects with a mean age of 45.8?±?8.39 years. There was no statistically significant difference between the groups in terms of age, sex and BMI. In the CSCR group the genotype frequencies were 4G/4G: 30% (n?=?18), 4G/5G: 50% (n?=?30), 5G/5G: 20% (n?=?12) and in the control group genotype frequencies were 34% (n?=?17), 42% (n?=?21) and 24% (n?=?12), respectively. There was no statistically significant difference in the distribution of genotypes among the groups (chi-squared, p?=?0.70). The CSCR group had a significantly higher serum PAI-1 concentration than the control group (p?=?0.001). In both groups the mean plasma PAI-1 concentration did not vary significantly among the different genotypes (p?>?0.05).

Discussion and conclusion: Although our results demonstrated that the patients with acute CSCR have higher serum PAI-1 concentrations than the controls, no significant difference was found in the genotype disturbances of the PAI-1 gene between the groups. The current study indicates that 4G/5G polymorphism in the promoter of the PAI-1 gene cannot be considered a risk factor for the elevated serum PAI-1 levels and consequent development of CSCR.     

Research on the comparison of the demethylvancomycin’s diffusion–deposition characteristics in the ocular solid tissues of sustained subtenon drug delivery with subconjunctival injection

Purpose: To compare the demethylvancomycin’s diffusion–deposition characteristics in the ocular solid tissues of sustained subtenon drug delivery with subconjunctival injection.

Method: Sixty adult white rabbits were randomly assigned to the subtenon drug delivery group and the subconjunctival injection group. The subtenon drug delivery group was continuously infused demethylvancomycin to the subtenon of rabbits. The subconjunctival injection group was injected demethylvancomycin to the subconjunctival of rabbits. Cornea, iris and sclera were collected for high-performance liquid chromatography analyses to determine drug concentrations at one hour, three hours, six hours, 12?h and 24?h of drug administration. WinNonlin 6.3 was used to calculate the parameters of cumulative area under the curve (AUC_cum) of demethylvancomycin.

Results: The peak levels of demethylvancomycin concentration of the subtenon drug delivery group and the subconjunctival injection group were 92.406?±?21.555 and 51.778?±?14.001?μg/g in cornea, 28.451?±?10.229?μg/g and 42.271?±?27.291?μg/g in iris, 153.166?±?51.738?μg/g and 57.423?±?18.480?μg/g in sclera. The differences of concentrations between the two groups in cornea and sclera were statistically significant (F?=?487.775, p?F?=?132.748, p?F?=?4.848, p?=?0.064). The maximum of AUC_cum of the subtenon drug delivery group and the subconjunctival injection group was 1808.23?h?*?μg/g and 273.73?h?*?μg/g in cornea, 489.12?h?*?μg/g and 216.16?h?*?μg/g in iris and 2166.34?h?*?μg/g and 392.57?h?*?μg/g in sclera at 24?h of drug administration.

Conclusion: The sustained subtenon drug delivery had a better drug permeability and accumulation in the intraocular solid tissue compared to subconjunctival injection, which demonstrated it was probably a promising and effective approach for treating posterior segment diseases and endophthalmitis.     

The effect of acute exposure to coarse particulate matter air pollution in a rural location on circulating endothelial progenitor cells: results from a randomized controlled study

Abstract

Context: Fine particulate matter (PM) air pollution has been associated with alterations in circulating endothelial progenitor cell (EPC) levels, which may be one mechanism whereby exposures promote cardiovascular diseases. However, the impact of coarse PM on EPCs is unknown.

Objective: We aimed to determine the effect of acute exposure to coarse concentrated ambient particles (CAP) on circulating EPC levels.

Methods: Thirty-two adults (25.9?±?6.6 years) were exposed to coarse CAP (76.2?±?51.5?μg?m^?3) in a rural location and filtered air (FA) for 2 h in a randomized double-blind crossover study. Peripheral venous blood was collected 2 and 20 h post-exposures for circulating EPC (n?=?21), white blood cell (n?=?24) and vascular endothelial growth factor (VEGF) (n?=?16–19) levels. The changes between exposures were compared by matched Wilcoxon signed-rank tests.

Results: Circulating EPC levels were elevated 2 [108.29 (6.24–249.71) EPC?mL^?1; median (25th–75th percentiles), p?=?0.052] and 20 h [106.86 (52.91–278.35) EPC?mL^?1, p?=?0.008] post-CAP exposure compared to the same time points following FA [38.47 (0.00–84.83) and 50.16 (0.00–104.79) EPC?mL^?1]. VEGF and white blood cell (WBC) levels did not differ between exposures.

Conclusions: Brief inhalation of coarse PM from a rural location elicited an increase in EPCs that persisted for at least 20 h. The underlying mechanism responsible may reflect a systemic reaction to an acute “endothelial injury” and/or a circulating EPC response to sympathetic nervous system activation.     

Atorvastatin affects C-reactive protein levels in patients with coronary artery disease

SUMMARY

Background: Elevated levels of C-reactive protein (CRP) are considered to be one of the indicators of poor prognosis in coronary artery disease (CAD). The aim of this study was to evaluate anti-inflammatory effects of atorvastatin in patients with CAD by measuring serum CRP levels.

Methods: After measuring the baseline levels of CRP and lipid fractions, the patients were divided into two groups. In Group A (n?=?46), atorvastatin (20?mg/day) was administered in addition to classic antianginal treatment (beta-blocker, nitrate and aspirin). In Group B (n?=?32), the usual antianginal treatment was continued. Following 4 weeks of treatment the same measurements were repeated.

Results: In Group A, CRP decreased from 20.3?mg/dl (95% CI, 9-31.8) to 10.8?mg/dl (95% CI, 2.7-18.9) (p?<?0.001). In Group B, CRP decreased from 17?mg/dl (95% CI, 13.1-21) to 12.8?mg/dl (95% CI, 9.7-15.9) (p?<?0.01). The decrease in group A was more than in group B (p?=?0.003).

Conclusions: In patients with CAD, atorvastatin exerted an anti-inflammatory effect represented by decreasing CRP levels. This effect was independent of the change in low density lipoprotein cholesterol (LDL-C) or high density lipoprotein cholesterol (HDL-C) levels.     

Comparison of intravitreal vancomycin and daptomycin application in experimental methicillin-resistant Staphylococcus aureus (MRSA) endophthalmitis in rabbits

Abstract

Purpose: To compare bactericidal activities of daptomycin and vancomycin in an experimental rabbit model of methicillin-resistant Staphylococcus aureus (MRSA) endophthalmitis.

Methods: The right eyes of 19 New Zealand rabbits weighing 2 to 2.5?kg were used. Each eye was inoculated with 1000 colony-forming units (cfu) of MRSA into the vitreous cavity. 24?h after the inoculation, the rabbits were randomly distributed into three groups: control group (n?=?5) was given 0.1?ml of balanced saline solution, daptomycin group 2 (n?=?7) was given 0.2?mg/0.1?ml daptomycin and vancomycin group 3 (n?=?7) was given 1?mg/0.1?ml vancomycin intravitreally. Clinical examination scores were recorded and vitreous aspirates were obtained for microbiological analysis on days 2 and 3 after MRSA inoculation. Rabbits were sacrificed, and the eyes were enucleated for histopathological examination.

Results: There was no difference between the daptomycin group, vancomycin group and control in terms of the clinical grading of endophthalmitis 24?h after the inoculation. In all treatment groups, mean number of cfu and histopathological scores were significantly lower compared to the control group. There was no difference between the daptomycin and vancomycin group in terms of the histopathological and clinical examination scores. Culture negativity achieved on day 3 was 71.4% and 57.1% in the daptomycin treatment group and the vancomycin treatment group, respectively.

Conclusions: Although both daptomycin and vancomycin are effective in treatment of experimental MRSA endophthalmitis, daptomycin has superior bactericidal activity 72?h after inoculation.     

Real world evidence of use of anti-VEGF therapy in Denmark

Objective: This study evaluates real-world evidence regarding the frequency of anti-vascular-endothelial-growth-factor (VEGF) injections during the first year of therapy of treatment-naïve patients with neovascular age-related macular degeneration (nAMD), diabetic macular edema (DME) and retinal vein occlusion (RVO) from the Danish National Patient Registry. There was a switch in anti-VEGF treatment for naïve nAMD patients during the study period, following the introduction of aflibercept, which was expected to reduce the injection frequency relative to ranibizumab due to a perception of prolonged treatment duration of aflibercept.

Methods: All treatment-naïve nAMD, DME or RVO patients who received an intravitreal injection in Denmark from 1 January 2012 to 31 July 2015 were eligible for inclusion. Patients were required to have been treated for at least one year and, for nAMD, to have received at least three injections during the first four months of treatment. Patients were allocated to half-year groupings (2012/1 to 2014/1) based on registration of their first intravitreal injection. Injection frequency during the first year of treatment was calculated for each group and t-tests investigated whether injection frequencies changed over time.

Results: In treatment naïve nAMD patients (n?=?500), the mean (SD) number of anti-VEGF injections increased significantly from 6.04 (1.71) in 2012/1 to 6.73 (1.62) in 2014/1 (p?=?.001; 2012/1 and 2012/2 vs. 2014/1) across all treatments. A similar trend was found for DME patients (n?=?76) from 2012/1 to 2014/1 and RVO patients (n?=?82) from 2012/2 to 2014/1, with mean injection frequencies increasing significantly from 5.14 (2.29) to 5.93 (1.98) (p?=?.007), and from 4.83 (1.21) to 6.08 (1.55) (p?=?.024), respectively. Post hoc sensitivity analysis also found a significant increase in injection frequency in nAMD patients who did not receive a loading phase (4.55 in 2012/1 and 5.05 in 2014/1; p?=?.006; n?=?616).

Conclusions: In contrast to the decrease in injection frequency predicted with a switch to aflibercept treatment for nAMD, our study showed that injection frequencies increased significantly from 2012 to 2014 in patients initiating therapy across the three diseases.     

Long-term intraocular pressure changes after intravitreal injection of bevacizumab

Purpose: To assess the long-term intraocular pressure (IOP) changes after the intravitreal injection of bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA) for treatment of age-related macular degeneration (AMD) and diabetic macular edema (DME) patients and evaluate the correlation factors.

Material and methods: Patients with neovascular AMD or DME underwent treat-and-extended anti-VEGF regimen in one eye and followed more than 12 months were enrolled in this study. We set three criteria of IOP elevation: (1) the IOP of the treated eye increased above the contralateral eye for at least two consecutive visits; (2) the IOP of the treated eye increased above the pre-injection IOP for at least two consecutive visits; (3) and the IOP of the treated eye increased more than 5?mmHg above the baseline IOP for at least two consecutive visits. We used mixed model univariate and multivariate analysis to assess the association between IOP elevation and independent parameters including age, sex, lens status, the number of injections, and underlying disease.

Results: In total 152 patients, 83 patients with AMD and 69 patients with DME, were included in this study. Mean follow-up time was 18.7 months, with a maximum of 50 months. In IOP elevation, 54 eyes (35.6%) showed an IOP increase above that of the contralateral eye (criteria 1), 50 eyes (33.4%) showed an IOP increase above the baseline IOP (criteria 2), and an IOP increase greater than 5?mmHg above the baseline IOP observed in nine eyes (5.9%) (criteria 3). In the univariate analysis, lens status and total number of injections were statistically significant for criteria 2 and 3 (all ps?<?0.05). However, in the multivariable analysis, only the number of intravitreal injections was statistically correlated with sustained IOP elevation for criteria 2 and 3 (p?<?0.001 and p?=?0.039, respectively).

Conclusions: Our results suggest that under long-term monitoring, with a treat-and-extended regimen, intravitreal bevacizumab injections were associated with sustained IOP elevation. In particular, multiple intravitreal injections could be associated with sustained IOP elevation.     

Systemic effects after intravitreal injection of bevacizumab in new born rabbit eyes

Purpose: To determine the systemic impact of intravitreal injection of bevacizumab (IVB), an anti-vascular endothelium growth factor antibody, in newborn rabbits.

Materials and methods: We used four groups of rabbits. Group 1 rabbits received a single injection of IVB starting from the age of 6?weeks. Group 2 rabbits received a single injection of balanced salt solution (BSS, 0.025?ml) and served as controls for group 1. Group 3 rabbits received two consecutive injections of IVB at the ages of 6 and 10?weeks. Group 4 rabbits received two consecutive injections of BSS at the ages of 6 and 10?weeks and served as controls for group 3. During the experiment, a complete blood count (CBC), clinical biochemistry, weight gain, food intake, body temperature, blood pressure, pulse, and mortality were measured in the animals. Two months after IVB injection, the animals were sacrificed, and histology of the major organs was checked. Immunohistochemistry was assessed to explore the neurons in the central nervous system (CNS).

Results: We found there were no morphological or functional changes in the eyes following IVB injection. Furthermore, there were no differences in CBC, biochemistry, or other measured parameters among the four groups of animals. We checked the histology of the major organs and neurons in the CNS and they did not reveal significant differences among the four groups of animals.

Conclusions: Conclusively, IVB of either one or two injections (0.625?mg) in newborn rabbit eyes is well tolerated and does not cause noticeable systemic organ pathology.     

Magnetic drug targeting in a rhabdomyosarcoma rat model using magnetite-dextran composite nanoparticle-bound mitoxantrone and 0.6 tesla extracorporeal magnets − sarcoma treatment in progress

Background: Magnetic drug targeting (MDT) is a new treatment principle for tumors. Passive MDT (pMDT) uses cytostatics coupled to ferromagnetic nanoparticles, whereas in active MDT (aMDT), extracorporeal magnets are additionally placed over the tumor area.

Purpose: Mitoxantrone-magnetite-dextran composite particles were used to assess the distribution and effect of MDT.

Methods: We conducted two trials with n?=?60 rats transfected with R₁H rhabdomyosarcoma cells. In the biodistribution trial (n?=?36) mitoxantrone concentrations in tumor tissue versus plasma were measured after one or two dose administration for aMDT, pMDT, and uncoupled mitoxantrone. The dose/effect trial (n?=?24) assessed change in tumor volume at day 1 and 7 days after administration of 4, 6, or 8 doses of mitoxantrone using aMDT.

Results: Mitoxantrone-magnetite-dextran concentration in blood was significantly (p?<?0.05) lower when using aMDT and as low as uncoupled mitoxantrone. Concentrations in tumor tissue were always significantly higher using MDT when compared to uncoupled mitoxantrone. Two doses resulted in drug accumulation inside the tumor. Tumor growth was significantly decreased with four doses using aMDT versus no treatment. Tumor size on day 8 versus day 1 was significantly (p?<?0.05) reduced after administration of six doses of mitoxantrone-magnetite-dextran. No allergies/toxic reactions were observed.

Conclusions: The MDT achieves higher levels of cytostatics in tumor tissue without increased systemic concentrations and succeeds in reducing tumor volume.     

Antihyperlipidemic activity of adenosine triphosphate in rabbits fed a high-fat diet and hyperlipidemic patients

Context Recently, adenosine triphosphate (ATP) was occasionally found to decrease the triglyceride (TG) levels in several hyperlipidemic patients in our clinical practice.

Objective The study investigates the anti-hyperlipidemic effects of ATP in a high-fat fed rabbit model and hyperlipidemic patients.

Materials and methods Twenty-four rabbits were randomly divided into three groups of eight animals each as follows: normal diet, high-fat diet and high-fat diet?+?ATP group. ATP supplementation (40?mg/day) was started at the 20th day and lasted for 10 days. Serum concentrations of total cholesterol (TC), TG, LDL-C, HDL-C were measured on the 20th day and 30th day. Heart, liver and aorta were subjected histopathological examination. Twenty outpatients diagnosed primary hyperlipidemia took ATP at a dose of 60?mg twice a day for 1 week.

Results Feeding rabbits with a high-fat diet resulted in a significant elevation of lipid parameters including TC, TG, LDL-C, VLDL-C compared to the normal diet group (p?<?0.01). ATP treatment significantly decreased serum TG level (p?<?0.01), whilst other parameters remained statistically unaltered. Meanwhile, ATP significantly reduced the thickness of fat layer in cardiac epicardium (p?<?0.05) and pathological gradation of ballooning degeneration in hepatocytes (p?<?0.05). After taking ATP for 1 week, hyperlipidemia patients exhibited a significant decrease of TG (p?<?0.01), but other lipid parameters had no significant change.

Discussion and conclusion The study indicates that ATP selectively decreases serum TG levels in high-fat diet rabbits and hyperlipidemic patients. Therefore, ATP supplementation may provide an effective approach to control TG level.