Assessment of real-time polymerase chain reaction detection of Acanthamoeba and prognosis determinants of Acanthamoeba keratitis.

OBJECTIVE: To evaluate the diagnostic value of real-time polymerase chain reaction (PCR) for detecting Acanthamoeba in eyes diagnosed with Acanthamoeba keratitis (AK) by conventional tests. In addition, to determine the preoperative prognosis-determining factors in eyes with AK. DESIGN: Retrospective, cross-sectional study. PARTICIPANTS: A total of 104 eyes of 103 patients who were diagnosed with AK or with bacterial or bacteria-associated keratitis (BK) by conventional tests. METHODS: Twenty-nine eyes with AK and 75 eyes with BK were evaluated for Acanthamoeba and bacterial DNA by real-time PCR. The Acanthamoeba copy numbers, bacterial load, and clinical parameters in the patients with AK were assessed for those significantly associated with poor outcome, that is, final visual acuity of <20/50 or requiring keratoplasty, by logistic regression analysis. MAIN OUTCOME MEASURES: Acanthamoeba DNA copy number, bacterial DNA copy number, and odds ratio (OR) for poor prognosis. RESULTS: The detection of amoebic DNA was 50 times more sensitive by real-time PCR than by conventional cyst counting. The Acanthamoeba copy numbers at the first visit (mean: 4.7×10(5)±3.2×10(5) copies) were significantly correlated with the AK stage, and both were significant risk factors for a poor outcome. The Acanthamoeba DNA copy numbers at the first visit and AK stage had a significantly high risk for poor outcome (OR of Acanthamoeba DNA copy per logarithm of copy numbers: 3.48, 95% confidence interval [CI], 1.04-111.63, P<0.05; OR of AK stage: 2.8 per stage increase, 95% CI, 1.07-7.30, P<0.05, after adjustment of age). In the AK cases with poor outcome, the amoebic DNA was not reduced by more than 90% after 1 month of treatment. The weak amoebic reduction was significantly associated with advanced AK stages or previous use of steroids. Bacterial 16S rDNA was detected in 53.6% of the eyes with AK, but it was not associated with any risk for refractoriness. CONCLUSIONS: Real-time PCR was effective in detecting and managing AK. The Acanthamoeba copy number and AK stage at the first visit were significantly associated with poor outcome. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Atypical Symptoms in Migraine-Related Alice in Wonderland Syndrome: Expansion of the Phenotype and Reflection on the Pathomechanism.

We report an 8-year-old girl who experienced daily episodes of visual and somesthetic distortion and was diagnosed with Alice in wonderland syndrome (AIWS). Ophthalmologic assessment revealed best-corrected visual acuity of 0.2 in both eyes, and bilateral constricted tubular or spiral visual fields. Augmented amplitude of visually evoked potentials was revealed, and treatment with lomerizine and valproate showed favorable effect on the visual/somesthetic distortion as well as the visual field and acuity. Psychogenic visual problems can co-exist with the typical sensory distortion in AIWS, similarly to the case of psychogenic pseudo-seizures in subjects with epilepsy. Otherwise, an ambiguous borderline between psychological and physical pathomechanisms in migraine may also be characteristic of the migraine-related AIWS.

A CASE OF UNILATERAL RETINAL PIGMENT EPITHELIUM DYSGENESIS.

PURPOSE: To report a case of unilateral retinal pigment epithelium dysgenesis. METHODS: An 8-year-old boy with a large grayish lesion and leopard-spot pattern lesion at its periphery in his left eye underwent fluorescein angiography, fundus autofluorescence imaging, optical coherence tomography, and electroretinography. RESULTS: Fluorescein angiography showed an area of geographic hyperfluorescence with a peripheral pattern of dark spots. The area of retinal pigment epithelial atrophy showed hypofluorescence on fundus autofluorescence. Optical coherence tomography of the left eye showed attenuation of the inner segment-outer segment junction and choroidal thinning. Single-flash electroretinography and scotopic electroretinography showed normal results and did not differ between both the eyes. The 30-Hz flicker test and photopic electroretinography showed a decrease in amplitude in the left eye. CONCLUSION: Unilateral retinal pigment epithelium dysgenesis is very rare, and its prognosis is still unknown. Careful follow-up of the patient seems to be essential.

Indoleamine 2,3-dioxygenase 1 in corneal endothelial cells limits herpes simplex virus type 1-induced acquired immune response.

BACKGROUND: Corneal endothelial cells are known to be targets of herpes simplex virus type 1 (HSV-1) infection; however, the pathogenesis of HSV infections of the endothelial cells has not been definitively determined. The purpose of this study was to examine an unrecognised strategy of corneal endothelial cells to protect themselves from HSV-1 infection. METHODS: Immortalised human corneal endothelial cells (HCEn) were infected with HSV-1. Based on the global transcriptional profile, the expression of indoleamine 2,3-dioxygenase 1 (IDO1) was determined using real-time PCR and western blots. To examine whether IDO1 has any antiviral role, we tested whether viral replication was affected by blocking the activity of IDO1. The immune modulatory role of IDO1 was analysed to determine whether IDO1 might contribute to modulating the recall responses of HSV-1-sensitised CD4(+) T cells. RESULTS: IDO1 was strongly expressed in HCEn cells after HSV-1 infection. IDO1 blockade did not significantly restrict viral transcription or replication, arguing against a previously recognised antiviral role for IDO1. When HCEn cells were examined for antigen-presenting function, HSV-1-primed HCEn cells stimulated the proliferation of allogeneic CD4(+) T cells and interleukin 10 (IL-10) secretion. When the recall response to HSV-1 was measured by the mixed lymphocyte reaction, the HCEn-stimulated CD4(+) T cells modulated and limited the recall response. When IDO1 was silenced in HCEn cells, the HCEn-mediated immune modulatory activity and regulatory T-cell activation were reduced. Overexpression of IDO1 promoted immune modulatory activity, which was partly conveyed by IL-10. CONCLUSIONS: IDO1 induced by HSV-1 infection limits and dampens excessive acquired immune responses in corneal endothelial cells.

Relationship between the number of cytomegalovirus in anterior chamber and severity of anterior segment inflammation.

PURPOSE: To characterize the cytomegalovirus-associated anterior segment inflammation and to determine whether the number of cytomegalovirus is significantly correlated with the disease characteristics. METHODS: Retrospective consecutive case series. Seventy-three patients with refractory anterior segment inflammation due to iridocyclitis, corneal endotheliitis and keratouveitis were studied. All the patients were suspected to have cytomegalovirus infection and had undergone real-time PCR of the aqueous humor to determine the amount of cytomegalovirus DNA. RESULTS: Cytomegalovirus DNA was detected in 24 of the 73 cases. The cytomegalovirus copy number was significantly correlated with the number of recurrent episodes and glaucoma treatment levels, but was not significantly correlated with the disease type. A high cytomegalovirus copy number was a significant risk factor for IOP elevation [Odds ratio (OR) per logarithm CMV amount: 2.5 (95 % confidence interval (CI) 1.1-5.4), presence of coin-shaped lesions (2.3 (1.3-4.0)), recurrent inflammation (2.1 (1.3-3.5)), and reduction of endothelial cell densities (1.7 (1.2-2.5))]. An IOP elevation [OR 18.2 (95 % CI 2.2-153.0)], reduction of endothelial cell densities [13.2 (2.9-60.0)], and recurrent inflammations [11.9 (2.5-56.6)], but not the disease type, were significant predictors of the presence of >10(3) copies/ml cytomegalovirus in the aqueous. CONCLUSIONS: Measurements of the cytomegalovirus DNA amount is useful for evaluating the severity of the anterior segment inflammation.