Expression profiling of the ephrin (EFN) and Eph receptor (EPH) family of genes in atherosclerosis-related human cells.

Ephrin B1 and its cognate receptor, Eph receptor B2, key regulators of embryogenesis, are expressed in human atherosclerotic plaque and inhibit adult human monocyte chemotaxis. Few data exist, however, regarding the gene expression profiles of the ephrin (EFN) and Eph receptor (EPH) family of genes in atherosclerosis-related human cells. Gene expression profiles were determined of all 21 members of this gene family in atherosclerosis-related cells by reverse transcription-polymerase chain reaction analysis. The following 17 members were detected in adult human peripheral blood monocytes: EFNA1 and EFNA3 - EFNA5 (coding for ephrins A1 and A3 - A5); EPHA1, EPHA2, EPHA4 - EPHA6 and EPHA8 (coding for Eph receptors A1, A2, A4 - A6 and A8); EFNB1 and EFNB2 (coding for ephrins B1 and B2); and EPHB1 - EPHB4 and EPHB6 (coding for Eph receptors B1 - B4 and B6). THP-1 monocytic cells, Jurkat T cells and adult arterial endothelial cells also expressed multiple EFN and EPH genes. These results indicate that a wide variety of ephrins and Eph receptors might affect monocyte chemotaxis, contributing to the development of atherosclerosis. Their pathological significance requires further study.

Progression from macular retinoschisis to retinal detachment in highly myopic eyes is associated with outer lamellar hole formation.

AIMS: To investigate the morphological changes that occur during the development of an early retinal detachment (RD) from a myopic macular retinoschisis (MRS) by optical coherence tomography (OCT). METHODS: The OCT images of five eyes of five consecutive patients with myopic MRS who developed an RD during the follow-up period were studied. RESULTS: The progression from MRS to early RDs went through four stages. In stage 1, OCT images appeared to show a focal irregularity of the thickness of external retina. In stage 2, an outer lamellar hole developed within the thickened area and a small RD developed. In stage 3, the column-like structures overlying the hole seemed to be separated horizontally, and the outer lamellar hole appeared to be larger vertically. In stage 4, the upper edge of the external retina was further elevated and attached to the upper part of the retinoschisis layer accompanied by further enlargement of the RD. CONCLUSIONS: This longitudinal study showed that the progression from myopic MRS to RD passes through four stages, and the formation of an outer lamellar hole predisposes the retina to a RD. These OCT findings might be useful for considering the surgical indication for eyes with a myopic MRS.

Use of multiplex PCR and real-time PCR to detect human herpes virus genome in ocular fluids of patients with uveitis.

AIM: To measure the genomic DNA of human herpes viruses (HHV) in the ocular fluids and to analyse the clinical relevance of HHV in uveitis. METHODS: After informed consent was obtained, a total of 111 ocular fluid samples (68 aqueous humour and 43 vitreous fluid samples) were collected from 100 patients with uveitis. The samples were assayed for HHV-DNA (HHV1-8) by using two different polymerase chain reaction (PCR) assays, qualitative PCR (multiplex PCR) and quantitative PCR (real-time PCR). RESULTS: In all of the patients with acute retinal necrosis (n = 16) that were tested, either the HSV1 (n = 2), HSV2 (n = 3), or VZV (n = 11) genome was detected. In all patients, high copy numbers of the viral DNA were also noted, indicating the presence of viral replication. In another 10 patients with anterior uveitis with iris atrophy, the VZV genome was detected. When using multiplex PCR, EBV-DNA was detected in 19 of 111 samples (17%). However, real-time PCR analysis of EBV-DNA indicated that there were only six of the 19 samples that had significantly high copy numbers. The cytomegalovirus (CMV) genome was detected in three patients with anterior uveitis of immunocompetent patients and in one immunocompromised CMV retinitis patient. In addition, one patient with severe unilateral panuveitis had a high copy number of HHV6-DNA. There was no HHV7- or HHV8-DNA detected in any of the samples. CONCLUSIONS: A qualitative multiplex PCR is useful in the screening of viral infections. However, the clinical relevance of the virus infection needs to be evaluated by quantitative real-time PCR.

P2Y2 receptor elevates intracellular calcium concentration in rabbit eye suprachoroid.

Effects of ATP on the intracellular free calcium ion concentration ([Ca2+]i) in the rabbit eye suprachoroid were investigated by means of fura-2 microfluorophotometry. ATP application (10 to 100 microM) elicited a dose-dependent biphasic [Ca2+]i-increase: a fast phase typically peaking within 30 s and a following slow plateau phase, which lasted during the presence of ATP. The slow plateau phase was markedly diminished by removal of extracellular Ca2+, whereas the fast phase remained. An inhibitor of Ca2+ release from the sarcoplasmic reticulum (TMB-8), an endoplasmic Ca2+-ATPase inhibitor (thapsigargin) and a phospholipase C (PLC) inhibitor (U-73122) diminished the fast phase. A P2 receptor antagonist (Suramin) inhibited the ATP-induced [Ca2+]i-response. The potency order of ATP and related substances in producing the [Ca2+]i-elevation was UTP approximately equals ATP>ATP-gamma-S>ITP>ADP. beta,gamma-MethyleneATP, 2-methylthioATP and UDP evoked no response. This order is consistent with the P2Y2 receptor characteristics. Cross-desensitization between ATP and UTP excludes the co-existence of the other types of receptors. In conclusion, the ATP-induced [Ca2+]i-elevation in the rabbit eye suprachoroid was elicited by the Ca2+ release from the PLC-dependent, thapsigargin-sensitive intracellular Ca2+ storage sites by activating P2Y2 nucleotide receptors.