Vitreous Fluid Samples Research Articles

Role of Intravitreal Inflammatory Cytokines and Angiogenic Factors in Proliferative Diabetic Retinopathy

Objective: Inflammatory reaction has been shown to involve the progress of type 2 (non-insulin-dependent) diabetes. We, therefore, examined the effects of inflammatory cytokines and angiogenic factors in the pathogenesis of proliferative diabetic retinopathy (PDR) in type 2 diabetes.Patients and methods: Vitreous fluid samples were obtained by vitrectomy from 62 eyes of PDR patients with type 2 diabetes and from 20 eyes of age-matched non-diabetic patients. The concentrations of interleukin 1 beta (IL1B), IL6, IL8, IL10, chemokine (C-C motif) ligand 2 (CCL2), endothelin 1 (EDN1), vascular endothelial growth factor (VEGF), and tumor necrosis factor (TNF) in the vitreous samples were measured by enzyme-linked immunosorbent assay (ELISA).Results: The concentrations of LI1B, IL6, IL8, CCL2, EDN1, VEGF, and TNF in the vitreous samples were considerably higher in PDR patients in comparison with the controls. However, the level of IL10 in PDR patients was similar to that obtained in the controls. Analysis of the correlations of the studied factors revealed the correlation of VEGF and IL6, VEGF and EDN1, IL8 and CCL2, and EDN1 and TNF in PDR patients. In addition, a significant positive correlation was observed between vitreous TNF as well as EDN1 and serum HbA1c levels in PDR patients.Conclusions: The inflammatory cytokines and angiogenic factors IL1B, IL6, IL8, CCL2, EDN1, VEGF, and TNF are increased in the vitreous of PDR patients without an increase in IL-10. These results add support to the role of inflammatory cytokines and angiogenic factors in the genesis of PDR. Understanding the implication of these cytokines may provide diagnostic tools and therapeutic targets for treatment and prevention of PDR.

Use of DNA microarray analysis in diagnosis of bacterial and fungal endophthalmitis

BackgroundTo examine the utility of DNA microarray analysis for identifying causative microorganisms in endophthalmitis.MethodsThirteen samples of vitreous fluid (VF) were obtained from 13 patients during vitrectomy. Vitreous fluids from three patients with suspected endophthalmitis and ten controls without infection were subjected to testing for the presence of bacteria and fungi in culture tests, polymerase chain reaction (PCR) analysis, and DNA microarray analysis.ResultsNo control sample was positive for bacteria or fungi in the culture test, PCR, or microarray analysis. Specimens from two patients (Cases 1 and 2) with suspected endophthalmitis were positive for bacteria in PCR, and a specimen from one patient (Case 3) was positive for fungi in PCR. Klebsiella pneumonia (Case 1), Streptococcus agalactiae (Case 2), and Candida parapsilosis (Case 3) in the PCR-positive specimens were identified by DNA microarray analysis within 24 hours. Culture results were also positive for K. pneumonia in Case 1, S. agalactiae in Case 2, and C. parapsilosis in Case 3, but required 3 to 4 days to obtain.ConclusionsMicroarray analysis is complementary to routine cultures for identifying causative microorganisms and is likely to be a useful tool in patients with suspected endophthalmitis who require rapid diagnosis and early antibiotic treatment.

Vascular Endothelial Growth Factor Receptor-2 in Macular Oedema with Retinal Vein Occlusion

Background: To evaluate the relationships among vitreous fluid levels of soluble vascular endothelial growth factor receptor-2 (sVEGFR-2), vascular endothelial growth factor (VEGF), and the severity of macular oedema secondary to branch retinal vein occlusion (BRVO). Methods: Twenty-five BRVO patients with macular oedema were enrolled. Vitreous fluid samples were obtained during vitreoretinal surgery to measure the levels of sVEGFR-2 and VEGF. Macular oedema was examined by optical coherence tomography. Results: The vitreous fluid levels of VEGF and sVEGFR-2 were significantly correlated with the severity of macular oedema (ρ = 0.54, p = 0.008 and ρ = 0.40, p = 0.047, respectively). The sVEGFR-2 × VEGF product was also significantly correlated with the severity of macular oedema (ρ = 0.62, p = 0.003). Conclusions: The severity of macular oedema was more closely associated with sVEGFR-2 × VEGF than sVEGFR-2 or VEGF alone, suggesting that macular oedema in BRVO patients may be influenced by both VEGF and sVEGFR-2.

VITREOUS INFLAMMATORY FACTORS AND SEROUS MACULAR DETACHMENT IN BRANCH RETINAL VEIN OCCLUSION

To investigate whether vascular endothelial growth factor, soluble intercellular adhesion molecule-1 (sICAM-1), and pigment epithelium-derived factor are associated with serous retinal detachment (SRD) secondary to branch retinal vein occlusion. The subjects were 44 branch retinal vein occlusion patients with macular edema and 16 controls. Patients were divided into 2 groups by optical coherence tomography findings, that is, 18 patients with SRD and 26 with cystoid macular edema. The area of capillary nonperfusion was measured with fluorescein angiography and Scion Image software. Vitreous fluid samples obtained during pars plana vitrectomy were examined by enzyme-linked immunosorbent assay. The incidence of major branch retinal vein occlusion was significantly higher in SRD patients (17/18, 94%) than in cystoid macular edema patients (15/26, 58%, P = 0.007), while the nonperfused retinal area was significantly larger in SRD patients (P = 0.006). Vitreous fluid levels of vascular endothelial growth factor and soluble intercellular adhesion molecule-1 (sICAM-1) showed a significant increase across the 3 groups (control group, cystoid macular edema group, and SRD group) (P trend < 0.001 and P trend < 0.001, respectively), while the pigment epithelium-derived factor level showed a significant decrease across the 3 groups (P trend < 0.001). An excessive increase of vascular permeability secondary to upregulation of vascular endothelial growth factor and soluble intercellular adhesion molecule-1 (sICAM-1) along with downregulation of pigment epithelium-derived factor may contribute to the development of SRD in BRVO patients.

Vascular Endothelial Growth Factor and Interleukin-6 are Correlated With Serous Retinal Detachment in Central Retinal Vein Occlusion

Purpose/Aim: To investigate aqueous humour and vitreous fluid levels of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) in patients with central retinal vein occlusion (CRVO) and serous retinal detachment.Materials and methods: In twenty patients (20 eyes) with CRVO and 8 patients (8 eyes) with non-ischaemic ocular disease as controls, optical coherence tomography showed that 10 CRVO patients had serous retinal detachment (subretinal fluid with low reflectivity) and 10 patients had cystoid macular oedema (hyporeflective intraretinal cavities). Retinal ischaemia was evaluated from the area of capillary nonperfusion on fluorescein angiograms. To measure VEGF and IL-6 levels, aqueous humour and vitreous fluid samples were obtained during combined pars plana vitrectomy and cataract surgery.Results: Ischaemia was significantly more frequent among the CRVO patients with serous retinal detachment than those with cystoid macular oedema (10/10 vs. 3/10, P = 0.001). Aqueous humour levels of VEGF and IL-6 were significantly higher in the serous patients than the cystoid patients (P = 0.004 and P < 0.001, respectively). Vitreous fluid levels of VEGF and IL-6 were also significantly higher in the serous patients than the cystoid patients (P = 0.007 and P < 0.001, respectively).Conclusions: These results suggest that a marked increase of vascular permeability due to upregulation of VEGF and IL-6 may contribute to serous retinal detachment in patients with CRVO.

Detection of Candida and Aspergillus species DNA using broad-range real-time PCR for fungal endophthalmitis

The goal of this work is to establish a broad-range real-time polymerase chain reaction (PCR) diagnostic system for ocular fungal infection and to measure Candida and Aspergillus DNA in the ocular fluids obtained from unknown uveitis/endophthalmitis patients. After obtaining informed consent, intraocular fluids (aqueous humor and vitreous fluid samples) were collected from 54 patients with idiopathic uveitis or endophthalmitis. Samples were assayed for Candida or Aspergillus DNA using broad-range (18S rRNA sequences) quantitative real-time PCR. Candida or Aspergillus DNA was detected in seven out of 54 patient ocular samples (13%). These PCR-positive samples showed significantly high copy numbers of Candida or Aspergillus DNA. On the other hand, fungal DNA was not detected in any of the other 46 samples collected from these idiopathic uveitis or endophthalmitis patients. In the one PCR-negative case, PCR did not detect any fungal genome in the sample, even though this patient was clinically suspected of having Candida endophthalmitis. Real-time PCR results were negative for fungal DNA in the bacterial endophthalmitis patients and in various uveitis patients. In addition, fungal DNA was also not detected in patients without ocular inflammation (controls). Analysis of ocular samples by this broad-range real-time PCR method can be utilized for rapid diagnosis of patients suffering from unknown intraocular disorders such as idiopathic uveitis/endophthalmitis.

Osteopontin expression in vitreous and proliferative retinal membranes of patients with proliferative vitreous retinopathy.

To analyze osteopontin (OPN) expression in vitreous and proliferative retinal membranes of patients with proliferative vitreous retinopathy (PVR). A total of 54 vitreous fluid samples were obtained between 2009 and 2010, which contained 45 with PVR (group A) and 9 without PVR (group B). Enzyme-linked immunosorbent assay was applied to quantify the OPN concentrations in vitreous fluid. Four samples of proliferative retinal membrane were also obtained at the time of vitrectomy, and their contents of OPN were measured by Real-time RT-PCR. The OPN levels in the vitreous fluid were 778.48±62.06ng/mL in group A and 452.99±32.52ng/mL in group B. The vitreous OPN levels in group A were significantly higher than those in group B and to rise by time in the early stages of PVR. The average OPN levels in the proliferative retinal membranes (F=0.14) were also higher than those in the retinal pigment cells (F=0) using Real-time RT-PCR. The high vitreous and proliferative retinal membrane OPN levels in PVR suggest that OPN might promote the development of PVR. The vitreous OPN concentrations are rising by the time in the early phases of PVR.

Soluble Vascular Endothelial Growth Factor Receptor-2 and Inflammatory Factors in Macular Edema with Branch Retinal Vein Occlusion

To investigate relationships among vitreous fluid levels of soluble vascular endothelial growth factor receptor-2 (sVEGFR-2), vascular endothelial growth factor (VEGF), and soluble intercellular adhesion molecule 1 (sICAM-1) in patients with branch retinal vein occlusion (BRVO) and macular edema or patients with idiopathic macular hole. Retrospective case-control study. Tokyo Women's Medical University and Eguchi Eye Hospital. Forty-nine Japanese patients who underwent unilateral vitrectomy (27 with BRVO and 22 with macular hole). Vitreous fluid samples were obtained during vitreoretinal surgery to measure the levels of sVEGFR-2, VEGF, and sICAM-1. Retinal ischemia was evaluated from capillary nonperfusion on fluorescein angiography. Macular edema was examined by optical coherence tomography. Vitreous fluid levels of the 3 molecules and severity of macular edema. BRVO patients had a significantly higher vitreous fluid level of sVEGFR-2 (median, 1670 pg/mL; interquartile range [IQL], 1205 to 2225 pg/mL) than macular hole patients (median, 1265 pg/mL; IQR, 731 to 1800 pg/mL; P = .017), as was the case for VEGF (median, 237 pg/mL; IQR, 42.2 to 1305 pg/mL; vs median, 15.6 pg/mL; IQR, 15.6 to 15.6 pg/mL; P < .001) and sICAM-1 (median, 10.1 ng/mL; IQR, 6.3 to 22.5 ng/mL; vs median, 4.1 ng/mL; IQR, 3.3 to 6.0 ng/mL; P < .001). In BRVO patients, there was a significant positive correlation between vitreous fluid levels of sVEGFR-2 or VEGF and sICAM-1, but not between sVEGFR-2 and VEGF. Vitreous fluid levels of all 3 molecules were correlated significantly the with severity of macular edema in BRVO patients. sVEGFR-2 may induce an increase of vascular permeability together with or via sICAM-1, or both with and via sICAM-1, in BRVO patients with macular edema.

Vitreous Vascular Endothelial Growth Factor Concentrations in Proliferative Diabetic Retinopathy versus Proliferative Vitreoretinopathy

Purpose: To assess vitreous vascular endothelial growth factor (VEGF) concentrations in proliferative diabetic retinopathy (PDR) in comparison to proliferative vitreoretinopathy (PVR). Patients and Methods: Vitreous samples were collected from 69 eyes of 69 patients with traumatic lens dislocation (n = 10), grade B PVR with rhegmatogenous retinal detachment (n = 13), grade C PVR with rhegmatogenous retinal detachment (n = 14), PDR with vitreous hemorrhage (n = 18), and PDR with vitreous hemorrhage and tractional retinal detachment (n = 14). Vitreous fluid samples were obtained at vitrectomy, and the levels of VEGF were measured by enzyme-linked immunosorbent assay. Results: The mean vitreous level of VEGF was 15.14 ± 5.22 pg/ml in eyes with grade B PVR, 99.15 ± 38.58 pg/ml in eyes with grade C PVR, 4,534.01 ± 1,193.28 pg/ml in eyes with vitreous hemorrhage secondary to PDR, 5,157.29 ± 969.44 pg/ml in eyes with vitreous hemorrhage and tractional retinal detachment secondary to PDR, and 16.19 ± 5.76 pg/ml in eyes of the control group with traumatic lens dislocation. Vitreous VEGF concentrations were significantly higher in the patients with grade C PVR, PDR with vitreous hemorrhage and PDR with vitreous hemorrhage and tractional retinal detachment in comparison to the control patients (p < 0.05). A significant alteration was not observed in patients with grade B PVR (p = 0.55). Conclusions: Vitreous VEGF concentrations are increased in PDR and grade C PVR. The high VEGF concentrations could suggest a possible effect of VEGF on advanced PVR.

Intravitreous high expression level of netrin-1 in patients with proliferative diabetic retinopathy.

To investigate the significance of netrin-1 and vascular endothelial growth factor (VEGF) in the pathogenesis of retinal angiogenesis, the levels of netrin-1 and VEGF in the vitreous fluid and serum of the proliferative diabetic retinopathy (PDR) and non-proliferative diabetic retinopathy (non-PDR) patients were measured. We then determined the netrin-1 and VEGF expression in the oxygen induced retinopathy (OIR) mice retinas. A total of 18 eyes from 18 patients were included in our study and 10 of them were collected from PDR patients and 8 from non-PDR patients. Undiluted vitreous fluid samples were collected during pars plana vitrectomy. Appropriate blood samples were collected if possible. Netrin-1 and VEGF levels in the vitreous fluid and plasma were determined by Enzyme-linked Immunosorbent Assays. OIR mice models were established, and netrin-1 and VEGF levels were determined by immunohistochemistry analysis. The levels of netrin-1 and VEGF in the vitreous of PDR patients were significantly higher than those in the controls (Mediannetrin-1=509.94 vs. 85.91 pg/ml, P<0.001 and Median VEGF=762.60 vs. 77.52 pg pg/ml, P<0.001). Netrin-1 was mainly expressed in GCL and INL of the retina in mice. Both netrin-1 and VEGF were up-regulated in OIR mice. Netrin-1 and VEGF levels were elevated in vitreous fluid of the PDR patients and the OIR mice retinas. Therefore, netrin-1 may play important roles in pathological retinal angiogenesis.

Expression profiles of cytokines and chemokines in vitreous fluid in diabetic retinopathy and central retinal vein occlusion

The involvement of cytokines and chemokines in vitreous fluid is important in the development and progression of diabetic retinopathy (DR) and central retinal vein occlusion (CRVO). In this study, the concentrations of cytokines and chemokines in the vitreous fluid of eyes with DR and CRVO were measured and compared. We studied 76 eyes with proliferative DR and diabetic macular edema (DR group), 10 eyes with CRVO (CRVO group), and 23 eyes with an epiretinal membrane and macular hole (control group), among a series of 160 eyes from which vitreous fluid samples were collected during vitrectomy. The vitreous fluid samples were collected by suction with a vitreous cutter at the initial stage of vitrectomy. Twenty-seven different cytokines and chemokines were measured simultaneously using an array system (Bio-Plex(®)) with beads combined with antibodies (Bio-Rad), as follows: interleukin (IL)-1β, IL-1 receptor agonist, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, eotaxin, basic fibroblast growth factor, granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF), interferon (IFN)-γ, interferon-inducible 10-kDa protein (IP-10), monocytochemotactic protein-1 (MCP-1), macrophage inflammatory protein-1 alpha (MIP-1α), MIP-1β, platelet-derived growth factor (PDGF)-BB, regulated upon activation, normal T cell expressed and secreted, tumor necrosis factor alpha (TNF-α), and vascular endothelial growth factor (VEGF). Compared to the control group, the levels of IL-6, IL-8, IL-10, IL-13, IP-10, MCP-1, MIP-1β, PDGF and VEGF in the vitreous fluid were significantly higher in the DR group, while the levels of IL-1β, IL-2, IL-5, IL-8, IL-9, IL-10, IL-12, IL-13, eotaxin, G-CSF, IFN-γ, IP-10, MCP-1, MIP-1β, TNF-α and VEGF were significantly higher in the CRVO group. Compared to the DR group, IL-2, IL-9, IL-12, MCP-1 and IFN-γ were significantly elevated in the CRVO group. Multivariate regression analysis revealed that among 6 factors correlated to VEGF in the DR group, IL-10 and IL-13 were more positively correlated and PDGF was most inversely correlated to VEGF. In addition to inflammatory cytokines and neurotrophic factors such as VEGF, anti-inflammatory cytokines such as IL-10 and IL-13 may be involved more in the pathogenesis of DR and CRVO than in other diseases; cytokines and chemokines may also be correlated to VEGF in the vitreous fluid. It is also suggested that the inflammatory reaction may be more activate in CRVO than in DR.

Vitreous Levels of Pigment Epithelium–Derived Factor and Vascular Endothelial Growth Factor in Macular Edema with Central Retinal Vein Occlusion

Purpose: To investigate whether pigment epithelium-derived factor (PEDF) or vascular endothelial growth factor (VEGF) influences macular edema in patients with central retinal vein occlusion (CRVO).Materials and Methods: In 27 patients who had CRVO with macular edema and 21 patients with nonischemic ocular diseases (control group), retinal ischemia was evaluated by measuring the area of capillary nonperfusion on fluorescein angiography and macular edema was examined by optical coherence tomography. Vitreous fluid samples were obtained during pars plana vitrectomy.Results: The vitreous level of VEGF was significantly higher in the CRVO patients than in the controls (median: 366 vs. 15.6 pg/ml, P < 0.0001), while the vitreous level of PEDF was significantly lower in the patients than in the controls (median: 17.5 vs. 28.4 ng/ml, P = 0.0298). Vitreous levels of VEGF were significantly higher in CRVO patients with retinal ischemia than in those without ischemia (P < 0.0001), while PEDF levels did not show a significant difference. Vitreous levels of VEGF and PEDF were related to the retinal thickness at the central fovea (P = 0.0059 and P = 0.0308, respectively).Conclusions: VEGF and PEDF may independently influence retinal vascular permeability in CRVO patients with macular edema.

Vitreous inflammatory factors and serous retinal detachment in central retinal vein occlusion: a case control series

BackgroundThis study investigated whether the vitreous fluid levels of soluble vascular endothelial growth factor receptor-2 (sVEGFR-2), pigment epithelium-derived factor (PEDF), and soluble intercellular adhesion molecule 1 (sICAM-1) were associated with the occurrence of serous retinal detachment (SRD) in patients with central retinal vein occlusion (CRVO).MethodsWe recruited 33 patients with CRVO and macular edema, as well as 18 controls with nonischemic ocular diseases. Eighteen of the 33 patients with CRVO showed SRD on optical coherence tomography of the macula (defined as subretinal accumulation of fluid with low reflectivity), while the other 15 patients only had cystoid macular edema (CME, defined as hyporeflective intraretinal cavities). Retinal ischemia was evaluated by measuring the area of capillary non-perfusion using fluorescein angiography and the public domain Scion Image program, while central macular thickness (CMT) was examined by optical coherence tomography. Vitreous fluid samples were obtained during pars plana vitrectomy and levels of the target molecules were measured by enzyme-linked immunosorbent assay.ResultsIschemia was significantly more common in the SRD group (17/18 patients) than in the CME group (5/15 patients) (P < 0.001). The vitreous fluid level of sICAM-1 increased significantly across the three groups from the control group (4.98 ± 1.73 ng/ml) to the CME group (15.4 ± 10.1 ng/ml) and the SRD group (27.1 ± 17.7 ng/ml) (ptrend< 0.001). The vitreous fluid level of sVEGFR-2 also showed a significant increase across the three groups (1083 ± 541 pg/ml, 1181 ± 522 pg/ml, and 1535 ± 617 pg/ml, respectively, ptrend = 0.019). On the other hand, the vitreous fluid level of PEDF showed a significant decrease across the three groups (56.4 ± 40.0 ng/ml, 24.3 ± 17.3 ng/ml, and 16.4 ± 12.6 ng/ml, respectively, ptrend< 0.001).ConclusionsHigher levels of inflammatory factors (sICAM-1 and sVEGFR-2) and lower levels of anti-inflammatory PEDF were observed in macular edema patients with SRD, suggesting that inflammation plays a key role in determining the severity of CRVO.

Diagnosis of tubercular uveitis by quantitative polymerase chain reaction

PurposeTo report the use of real-time or quantitative polymerase chain reaction (qPCR) in confirming the diagnosis of tubercular uveitis.MethodsA qPCR assay using primers targeting the MPB64 gene of Mycobacterium tuberculosis (MTB) was developed. A standard curve for quantification of mycobacterial load was generated. Vitreous fluid samples from patients presumed to have tubercular uveitis were assayed to identify and quantify the mycobacterial load. The amount of the test sample product was interpolated from the standard curve of cycle threshold values generated from known starting copy number of standard MTB DNA.ResultsThe copies of MTB genomes in the three test samples studied were found to be 7.61 × 104, 4.53 × 104, and 1.43 × 105, respectively.ConclusionqPCR detected and quantified MTB genome from the vitreous fluid of eyes clinically suspected with tubercular uveitis.

Stromal Cell-Derived Factor-1 Is Essential for Photoreceptor Cell Protection in Retinal Detachment

Stromal cell-derived factor-1 (SDF-1) causes chemotaxis of CXCR4-expressing bone marrow-derived cells. SDF-1 is involved in the pathogenesis of various vascular diseases, including those of the eye. However, the role of SDF-1 in neuronal diseases is not completely understood. Here, we show higher SDF-1 levels in the vitreous humor of patients with retinal detachment (RD) compared with normal patients. SDF-1 correlated positively with the duration as well as the extent of RD. Furthermore, SDF-1 correlated significantly with levels of interleukin-6 and interleukin-8, but not with vascular endothelial growth factor. Western blot analysis results showed significant SDF-1 up-regulation in detached rat retinas compared with normal animals. Immunohistochemistry data showed that SDF-1 was co-localized with the glial cells of the detached retina. SDF-1 blockade with a neutralizing antibody increased photoreceptor cell loss and macrophage accumulation in the subretinal space. The retinal precursor cell line R28 expressed CXCR4. SDF-1 rescued serum starvation-induced apoptosis in R28 cells and enhanced their ability to participate in wound closure in a scratch assay. Our results indicate a surprising, protective role for SDF-1 in RD. This effect may be mediated directly or indirectly through other cell types.

Pigment epithelium-derived factor and vascular endothelial growth factor in branch retinal vein occlusion with macular edema

We investigated whether pigment epithelium-derived factor (PEDF) or vascular endothelial growth factor (VEGF) influence macular edema in patients with branch retinal vein occlusion (BRVO). This investigation aimed to clarify the influence of PEDF in the vitreous fluid on retinal vascular permeability in patients with macular edema secondary to BRVO. The findings were expected to be useful for the treatment of macular edema in BRVO patients. This was a retrospective cross-sectional comparative case series. Thirty-three BRVO patients with macular edema and 24 control patients with nonischemic ocular diseases were enrolled. Retinal ischemia was evaluated by measuring the area of capillary nonperfusion on fluorescein angiography with Scion Image software. Macular edema was examined by optical coherence tomography. Vitreous fluid samples were obtained via pars plana vitrectomy, and the VEGF and PEDF levels were determined by enzyme-linked immunosorbent assay. The vitreous level of VEGF was significantly higher in BRVO patients than controls (P < 0.001). The vitreous PEDF level was significantly lower in BRVO patients than controls (P = 0.026). In BRVO patients, vitreous levels of PEDF and VEGF showed a significant negative correlation with each other (P < 0.001). Additionally, the vitreous VEGF level had a significant positive correlation (P < 0.001) and the vitreous PEDF level had a significant negative correlation (P < 0.001) with the nonperfused retinal area in BRVO patients. Furthermore, vitreous levels of VEGF and PEDF showed significant positive (P = 0.001) and negative (P = 0.014) correlations, respectively, with macular edema in BRVO patients. VEGF and PEDF may inversely influence retinal vascular permeability in patients with ischemic BRVO and macular edema. However, prospective validation will be needed to confirm these observations.

Diagnosis of bacterial endophthalmitis by broad-range quantitative PCR

AimTo measure the bacterial genome in ocular fluids and to analyse the clinical relevance of infectious endophthalmitis.MethodsNineteen ocular fluid samples (eight aqueous humour and 11 vitreous fluid samples) were collected...

Human herpesvirus 6‐A, 6‐B, and 7 in vitreous fluid samples

Human herpesvirus 6 and 7 (HHV-6, HHV-7) have been associated with several neurologic syndromes and have been detected in nervous tissue from healthy persons; however, only two cases of HHV-6A have been reported to be associated with intraocular inflammatory disease. Vitreous fluid was tested from 101 patients, including 69 samples from patients with ocular inflammation including CMV retinitis, idiopathic retinitis, iritis, and vitritis, for HHV-6A, HHV-6B, and HHV-7 DNA by PCR. HHV-6A DNA (4,950 copies per ml) was detected in vitreous fluid from one patient with CMV retinitis, HHV-6B DNA (10,140 copies per ml) was detected in vitreous fluid from one patient with idiopathic ocular inflammation in the absence of CMV DNA, and HHV-7 was not detected in any of the vitreous samples. HHV-6A, HHV-6B, and HHV-7 DNA are detectable in less than 2% of vitreous samples in patients with ocular inflammation.

Intravitreous Vascular Endothelial Growth Factor and Hypoxia-Inducible Factor 1a in Patients With Proliferative Diabetic Retinopathy

To determine vascular endothelial growth factor (VEGF) and hypoxia-inducible factor 1a (HIF-1a) in the vitreous fluid of patients with proliferative diabetic retinopathy (PDR). Observational case-control study. Serum and vitreous fluid samples were obtained during vitrectomy from 42 eyes of diabetic patients with PDR (17 type I and 25 type II diabetes mellitus) and from 23 eyes of nondiabetic patients. Retinopathy of 19 diabetic patients was graded as active and the other 23 graded as quiescent according to retinal neovascularization. VEGF and HIF-1a of serum and vitreous fluid samples were measured by enzyme-linked immunosorbent assay. Both intravitreous VEGF and HIF-1a were higher in diabetic patients with PDR than in control subjects (P < .01 and P < .01, respectively). After adjusting for total intravitreous protein (TP) concentration, intravitreous VEGF/TP and HIF-1a/TP in diabetic patients remained significantly higher too. Correlations between intravitreous VEGF and HIF-1a and intravitreous VEGF/TP and HIF-1a/TP were observed in diabetic patients with PDR (r = 0.730; P < .01; r = 0.531; P < .01, respectively) but not in control subjects. Both intravitreous VEGF and HIF-1a were higher in diabetic patients with active PDR than in those with quiescent PDR (P < .01 and P < .05, respectively). Intravitreous VEGF/TP in patients with active PDR remained higher (P < .05). Correlation between intravitreous VEGF and HIF-1a, as well as correlation between intravitreous VEGF/TP and HIF-1a/TP, were observed in diabetic patients with active PDR and those with quiescent PDR but not observed in control subjects. Intravitreous VEGF and HIF-1a in diabetic patients with PDR are increased and related mutually. VEGF and HIF-1a, especially VEGF, are associated with the angiogenesis of PDR.

Local and genetical determinants of vascular endothelial growth factor in vision threating proliferative diabetic retinopathy

Abstract Purpose To search for the association between the –634 C/G polymorphism of the VEGF gene and vision threating proliferative diabetic retinopathy (PDR) and to determine whether vitreous levels of VEGF were affected by genetic factors. Methods In this cross sectional case‐control study 349 unrelated Slovene subjects (Caucasians) with type 2 diabetes mellitus were enrolled: 206 patients with vision threating PDR in whom vitrectomy was performed, and the control group of 143 subjects with type 2 diabetes of duration of more than 10 years who had no clinical signs of diabetic retinopathy. In 68 out of 206 patients with PDR (71 eyes) vitreous fluid samples (0.3 ml) were obtained by vitreoretinal surgery. Results The –634 C/G VEGF gene polymorphism was not associated with PDR. Moreover, significantly higher vitreous levels of VEGF were demonstrated in diabetics with the CC genotype compared to those with the other (CG + GG) genotypes. Conclusion Despite the effect of the –634 C/G VEGF gene polymorphism on vitreous levels of VEGF in PDR, it failed to contribute to the genetic susceptibility to PDR.