Corneal hyperalgesia in patients with short tear film break-up time dry eye.

PURPOSE: To evaluate corneal tactile and pain sensations in patients with short tear film break-up time dry eye (sBUT DE). METHODS: This study enrolled 60 patients with sBUT DE and 46 healthy volunteers from Japan. We evaluated corneal tactile and pain sensations using a modified method with the Cochet-Bonnet corneal esthesiometer. RESULTS: Patients with sBUT DE had higher corneal pain sensitivity (26.3 ±â€¯23.1 mm) than healthy subjects (6.9 ±â€¯16.4 mm), but similar corneal tactile sensation (52.0 ±â€¯15.5 mm and 52.9 ±â€¯14.9 mm, respectively). In patients with sBUT DE and corneal hyperalgesia (n = 22, 36.7%), defined as a pain sensitivity ≥40 mm (i.e., the cutoff value at the 95th percentile of corneal pain sensitivity in healthy subjects), a strong significant correlation was found between the subjective pain score and objective corneal pain sensation (R = 0.79). However, for the entire cohort, we found a weak positive correlation between the subjective pain score and objective corneal pain sensation. CONCLUSIONS: Patients with sBUT DE were hypersensitive to corneal pain, which suggested that corneal hyperalgesia partly accounted for subjective symptoms in patients with sBUT DE.

Multicentre clinical study of the herpes simplex virus immunochromatographic assay kit for the diagnosis of herpetic epithelial keratitis.

BACKGROUND/AIMS: The novel immunochromatographic assay (ICGA) kit was recently developed to diagnose herpes simplex virus (HSV) infection. This multicentre study aimed to evaluate the value of the ICGA kit for the diagnosis of herpetic epithelial keratitis by comparing it with immunofluorescence assay (IFA) and real-time PCR. METHODS: Corneal scrapings were collected from 117 patients, including 77 with herpetic keratitis as their final clinical diagnosis as well as 40 others at 21 facilities. These samples were tested by the ICGA kit, IFA and real-time PCR. RESULTS: The positive concordance between final clinical diagnosis and ICGA was 46.7% (35/75 cases) and the negative concordance was 100% (39/39). The positive and negative concordance between real-time PCR and ICGA were 57.4% (35/61 cases) and 100% (53/53), respectively. The positive and negative concordance between IFA and ICGA were 61.1% (22/36 cases) and 83.3% (55/66), respectively. In 92 cases where anti-HSV drugs were not prescribed prior to corneal scraping, the positive and negative concordance between final clinical diagnosis and ICGA were 55.0% (33/60 cases) and 100% (32/32), respectively. CONCLUSIONS: The ICGA kit has moderate sensitivity and high specificity, indicating clinical utility in the diagnosis of herpetic epithelial keratitis.

The relation between visual performance and clinical ocular manifestations in Stevens-Johnson syndrome.

PURPOSE: To investigate the relation between visual function, clinical findings, and visual symptoms in Stevens-Johnson syndrome (SJS) and to compare the results with Sjögren syndrome (SS) patients and normal subjects. DESIGN: Cross-sectional comparative study. METHODS: One hundred fifteen eyes of 59 consecutive patients with SJS and toxic epidermal necrolysis (TEN), 208 eyes of 104 healthy normal subjects, and 132 eyes of 66 SS patients were investigated in this multicenter study. All study subjects underwent tear function and ocular surface examinations, Landolt and functional visual acuity examinations, and the Japanese version of the NEI VFQ-25 (National Eye Institute Visual Function Questionnaire). RESULTS: The mean ocular surface grading scores were significantly higher and the mean score of all 12 NEI VFQ subscales was significantly lower in the SJS patients compared to the SS patients and the normal subjects (P < .05). The conventional and functional logarithm of minimal angle of resolution (logMAR) visual acuities in SJS patients with minimal corneal complications were significantly higher and the mean total composite NEI VFQ scores were lower compared to SS patients. The conventional and functional logMAR visual acuities and the mean ocular surface grading scores in SJS with aqueous deficiency were significantly higher and the mean total composite NEI VFQ scores were lower compared to SS patients. Strong correlations between best-corrected logMAR functional visual acuities and either ocular surface grading scores or the composite NEI VFQ-25 scores were observed. CONCLUSIONS: The functional visual acuity examination reflects the severity of clinical ocular surface findings and vision-related quality of life more than the standard conventional visual acuity in SJS.

[Survey of severe contact lens-associated microbial keratitis in Japan].

PURPOSE: To understand the current state of severe contact lens (CL)-associated microbial keratitis in Japan. METHOD: The survey was conducted by the Japan Contact Lens Society and the Japanese Association for Ocular Infection in 224 facilities from April 2007 to March 2009. Patients who were diagnosed with CL-associated microbial keratitis and hospitalized for treatment were enrolled. Clinical characteristics of the keratitis, microbiologic findings and the status of CL hygiene were studied. RESULTS: A total of 350 patients were investigated, with an average age of 28.0 (9-90) years. Acanthamoeba was identified in 85 (24.3%) corneal specimens and Pseudomonus aeruginosa in 70 (20.0%) cases. One hundred ninety six (56.0%) patients were frequent replacement soft CL users. Extended wearing of daily-use CLs was found in 77 (22.0%) patients. Only 67 cases maintained good CL hygiene by daily rubbing-washing and the poor CL care situation was reviewed. CONCLUSION: The most frequently detected pathogenic microorganism was Acanthamoeba, followed by Pseudomonus aeruginosa. Our survey showed the importance of keeping good CL hygiene by proper lens care, and improvement of CL-related social regulations is urgently needed.

Clinical features of adenoviral conjunctivitis at the early stage of infection.

PURPOSE: Because adenoviral conjunctivitis is a contagious disease, prompt and accurate diagnosis in the early stage of infection is necessary to prevent epidemics. We evaluated and compared the clinical features of adenoviral conjunctivitis at the first medical examination with those of nonadenoviral follicular conjunctivitis. METHODS: The clinical features of 102 patients with suspected adenoviral conjunctivitis at the first medical examination were retrospectively reviewed. Human adenovirus (HAdV) DNA in samples from the patients was detected by polymerase chain reaction, and HAdV DNA-positive and HAdV DNA-negative patients were respectively assigned to adenoviral and nonadenoviral follicular conjunctivitis groups. The two groups were compared for bilaterality, intrafamilial infection, multiple subepithelial corneal infiltrates (MSI), preauricular lymphadenopathy, and severity of conjunctivitis. RESULTS: Adenoviral conjunctivitis and nonadenoviral conjunctivitis were diagnosed in 68 and 34 patients, respectively. Bilaterality, intrafamilial infection, and MSI showed significant intergroup differences. Remarkably, MSI was observed in 42.6% of the patients in the early stage of infection. There were no significant intergroup differences in preauricular lymphadenopathy or severity of conjunctivitis at any stage. CONCLUSIONS: To accurately diagnose adenoviral conjunctivitis in the early stage, bilateral conjunctival conditions, history of intrafamilial infection, and MSI should be checked, even in cases of mild or moderate follicular conjunctivitis.

Molecular epidemiology of adenoviral keratoconjunctivitis in Saudi Arabia.

PURPOSE: Adenoviral keratoconjunctivitis is a major cause of ocular morbidity and may lead to visual loss. Adenovirus types 8, 19, and 37 may cause epidemic keratoconjunctivitis. The main objective of this study was to determine the types of adenoviruses causing keratoconjunctivitis in Saudi Arabia. METHODS: We conducted a non-interventional observational clinical study. Seventy three eyes from 65 patients who presented to The Eye Center in Riyadh, Saudi Arabia with clinical features of acute adenoviral keratoconjunctivitis were included. Each patient underwent complete clinical examination and features such as membranous reaction, conjunctival hemorrhage, subepithelial corneal infiltrates, and preauricular lymph node enlargement were recorded. Conjunctival swabs were obtained from patients with presumed acute viral conjunctivitis. Immunochromatography (IC) and restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) were performed on the conjunctival swabs obtained from each eye. Serotype identification was performed using direct sequencing technique. RESULTS: Forty-nine (67.1%) were adenovirus type 8, 8 (11.0%) were adenovirus type 3, 6 (8.2%) type 37, 5 (6.8%) were adenovirus type 4, and 2 (2.3%) type 19. The remaining 5 were types 14, 19, and 22. The prevalence of membranous conjunctivitis was highest (83%) among eyes with adenovirus type 37 while subepithelial corneal opacities were most commonly seen among eyes with adenovirus type 8 (47%). Immunochromatography tests were positive for adenovirus in 48 (65.7%) out of 73 eyes. CONCLUSIONS: This study determined the types of adenoviruses causing keratoconjunctivitis at one center in Saudi Arabia. Direct sequencing techniques is an efficient, accurate, and rapid means of diagnosing adenoviral keratoconjunctivitis. The most common causes of adenoviral keratoconjunctivitis in Saudi Arabia were adenovirus types 8, 3, and 37. Membranous conjunctivitis and subepithelial opacities had the highest frequency of adenovirus types 37 and 8, respectively. Lymph nodes enlargement was least likely in adenovirus type 4.

Nucleotide sequence variation in the hexon gene of human adenovirus type 8 and 37 strains from epidemic keratoconjunctivitis patients in Japan.

Human adenovirus type 8 (HAdV-8) and 37 (HAdV-37) cause epidemic keratoconjunctivitis (EKC) associated with community-acquired and nosocomial infections. The nucleotide sequences of the entire hexon and fiber genes of eight HAdV-8 and 26 HAdV-37 strains were analysed and the transition mutations in each gene were compared among strains. Compared with prototype strains, the hexon gene of HAdV-8 and -37 strains showed between two and seven and one and twelve variations at nine and 21 different positions, respectively. All of these, except one position in HAdV-37, were located in the conserved region 4 (C4). There were only three polymorphisms in the fiber gene of both HAdV-8 and HAdV-37, fewer than those in C4. The nucleotide sequence of HAdV-8 and -37 C4 might be readily modified during EKC epidemics.

Analysis of the complete genome sequence of epidemic keratoconjunctivitis-related human adenovirus type 8, 19, 37 and a novel serotype.

We determined the complete genome sequence of epidemic keratoconjunctivitis (EKC)-related human adenoviruses (HAdVs). We analysed a total of 12 HAdV strains; three prototype strains and two HAdV-8, three HAdV-19 and three HAdV-37 clinical isolates from EKC patients in Japan, and one novel serotype of HAdV. Genome organization of these serotypes was identical to those of the recently determined HAdV-19 and HAdV-37. The identities of the whole genome were over 99 % among strains from the same serotype, except for HAdV-19p, which is not associated with conjunctivitis, resulting in the formation of a distinct cluster in the phylogenetic analysis. The penton, loop 1 and loop 2 of hexon, early region 3 (E3) and fiber were hypervariable regions between serotypes. Results suggest that the HAdV-19 clinical strain is a recombinant of HAdV-19p-like and HAdV-37-like strains, and that the acquisition of the penton, E3 or fiber may be related to ocular tropism.

Epidemic keratoconjunctivitis due to the novel hexon-chimeric-intermediate 22,37/H8 human adenovirus.

In a 2-month period in 2003, we encountered an outbreak of epidemic keratoconjunctivitis (EKC) in Japan. We detected 67 human adenoviruses (HAdVs) by PCR from eye swabs of patients with EKC at five eye clinics in different parts of Japan. Forty-one of the 67 HAdV DNAs from the swabs were identified as HAdV-37 by phylogenetic analysis using a partial hexon gene sequence. When the restriction patterns of these viral genomes were compared with that of the HAdV-37 prototype strain, one isolate showed a never-before-seen restriction pattern. Within 1 year, we encountered three more EKC cases caused by a genetically identical virus: two nosocomial infections at two different university hospitals and a sporadic infection at an eye clinic. We determined the nucleotide sequences of the full-length hexon and fiber genes of these isolates and compared them to those of the 51 prototype strains. Surprisingly, the sequence of the hexon (epsilon determinant) loop-1 and -2 regions showed the highest nucleotide identity with HAdV-22, a rare EKC isolate. However, the nucleotide sequence of the fiber gene was identical to that of the HAdV-8 prototype strain. 22 We propose that this virus is a new hexon-chimeric intermediate HAdV-22,37/H8, and may be an etiological agent of EKC.

Novel human adenovirus causing nosocomial epidemic keratoconjunctivitis.

In 2000, we encountered cases of nosocomial infections with epidemic keratoconjunctivitis (EKC) at a university hospital in Kobe, in the western part of Japan. Two human adenovirus (HAdV) strains, Kobe-H and Kobe-S, were isolated from patients with nosocomial EKC infection. They were untypeable by existing neutralizing antisera; however, the isolate was neutralized with homologous antisera. We then encountered several cases of EKC due to nosocomial infections in eye clinics in different parts of Japan. A total of 80 HAdVs were isolated from patients with EKC at eight different hospitals. The partial hexon gene sequences of the isolates were determined and compared to those of the prototype strains of 51 serotypes. All isolates had identical partial hexon nucleotide sequences. Phylogenetic analysis classified these isolates into species of HAdV-D. The isolates showed 93.9 to 96.7% nucleotide identity with HAdV-D prototype strains, while all 32 HAdV-D prototype strains ranged from 93.2 to 99.2% identity. The sequences of the loop 2 and fiber knob regions from the representative strain, Kobe-H, were dissimilar in all prototype strains of 51 serotypes. We believe that this virus is a novel serotype of HAdV that causes EKC.

[The study of prevalence of neutralization antibody against human adenovirus in ophthalmological medical workers].

PURPOSE: We studied the prevalence of neutralization antibodies against human adenovirus (hADV) in ophthalmological medical workers as one measure for the prevention of nosocomial infection. METHODS: The prevalence of neutralization antibodies against hADV-3, -4, -7, -8, -11, -19, and -37, which can cause conjunctivitis, was measured in 288 workers at ten ophthamological facilities in Japan. We studied the prevalence in different facilities, different generations, different types of workers (doctors, nurses, and others), and their medical history of hADV conjunctivitis. RESULTS: Among the workers, the prevalence of neutralization antibodies against hADV-3, -4, -7, -8, -11, -19, and -37 was 70.1%, 43.8%, 18.8%, 16.3%, 16.3%, 8.7%, and 6.3%, respectively. The prevalence of neutralization antibodies against hADV-8 was two times higher in doctors than in other workers. People who have a history of hADV conjunctivitis have a high prevalence of neutralization antibodies against hADV-8, -19, and-37. CONCLUSIONS: The prevalence of neutralization antibodies against hADV-8, -19, and-37 was low in the ophthalmological medical workers. These seroepidemiological data indicate the high possibility of an epidemic of conjunctivitis and occurrence of nosocomial infection caused by these serotypes.

Five new genome types of adenovirus type 37 caused epidemic keratoconjunctivitis in Sapporo, Japan, for more than 10 years.

Human adenovirus type 37 (HAdV-37) is a major cause of epidemic keratoconjunctivitis and has recently been the largest causative agent of keratoconjunctivitis in Japan. To investigate the genetic characteristics of HAdV-37 strains isolated in Sapporo, we analyzed the genome types and genetic relationships of 51 strains isolated there from 1990 through 2001. By using DNA restriction analysis, eight genome types (HAdV-37/D1, HAdV-37/D3, and HAdV-37/D6 to HAdV-37/D11) were identified, including five new ones. The restriction fragments of these genome types shared more than 95% identity with those of the prototype strain. By DNA sequence analysis, five and three single nucleotide substitutions, respectively, were found in partial sequences of the hexon and fiber genes. The combinations of mutations resulted in four hexon and fiber types (hx1 to hx4 and f1 to f4) and six hexon/fiber pairs (hx1/f1, hx2/f1, hx1/f2, hx1/f3, hx3/f4, and hx4/f4). The six pairs correlated well with certain genome types. In all three epidemics of keratoconjunctivitis to strike Sapporo in the past 12 years, specific genome types and fiber types were usually isolated: in the first epidemic, HAdV-37/D1 (f1) and HAdV-37/D3 (f1); in the second, HAdV-37/D6 (f2) and HAdV-37/D8 (f3); and in the third, HAdV-37/D10 (f4) and HAdV-37/D11 (f4). We conclude that mutations in the adenovirus genome occurred chronologically and that certain mutations were correlated with the epidemics of adenoviral keratoconjunctivitis.

New genome type of adenovirus serotype 4 caused nosocomial infections associated with epidemic conjunctivitis in Japan.

Human adenovirus type 4 is one of the major serotypes isolated from patients with adenoviral conjunctivitis. In 2001 we encountered nosocomial infections with epidemic conjunctivitis in the ophthalmology ward of one hospital in Sapporo, which is in the northern part of Japan. Adenoviruses were isolated from the patients with this nosocomial infection and identified as adenovirus type 4 (AdV-4) by a neutralization test with serotype-specific antiserum. When the cleavage patterns of the isolates were compared with the full viral genome with BamHI and SmaI, the cleavage patterns of the isolates were shown to be different from those of AdV-4p and other previously known AdV-4 variants. The nucleotide sequences of the fiber gene of the isolates showed the highest homologies (94.3%) with AdV-4 among the nucleotide sequences available from GenBank and formed a monophyletic cluster along with the prototype strain of AdV-4. The isolates, however, were located in a different lineage from those of AdV-4p and the AdV-4 variant from the sporadic infections. We conclude that the nosocomial infection that appeared in 2001 was caused by a new genome type of AdV-4, which was designated AdV-4c.

Molecular diagnosis of human adenoviruses d and e by a phylogeny-based classification method using a partial hexon sequence.

Human adenoviruses (HAdVs) are the major causes of a variety of acute illnesses. Virus isolation and neutralization tests are usually done to identify the causative virus, but these tests are labor-intensive and time-consuming, and standardized antisera are in limited supply. This study investigated a rapid and reliable method of virus identification based on PCR and phylogenetic analysis. The phylogenetic tree constructed by neighbor joining on the basis of the newly determined partial hexon sequences from 33 prototypes of HAdV-D and -E, along with 11 available prototypes of HAdV-A to -C and -F from GenBank, allowed HAdVs to be grouped into six distinct clusters. These clusters correspond closely to the six newly designated species, HAdV-A to -F. The partial hexon sequences of 57 isolates from patients with acute conjunctivitis obtained over 20 years plus those of 44 prototype strains were analyzed. Each isolate formed a monophyletic cluster along with its respective prototype strain, allowing serotype identification. Partial-hexon-based classification appears to be an effective tool for studying the molecular epidemiology of HAdVs.

The presence of macrophage migration inhibitory factor in human trabecular meshwork and its upregulatory effects on the T helper 1 cytokine.

PURPOSE: To investigate the expression and secretion of macrophage migration inhibitory factor (MIF) in human trabecular meshwork (HTM) and evaluate its role in ocular inflammation. METHODS: Tissue samples of HTM cells were isolated from donor human eyes or corneoscleral buttons, and the HTM cells were cultured. The expression of MIF on HTM cells was evaluated by RT-PCR, Western blot analysis, and ELISA. T-cell clones (TCCs) were established from ocular infiltrating cells of patients with uveitis. ELISA was used to evaluate the pathologic role of MIF, in relation to regulatory effects on cytokine production by T cells. RESULTS: MIF was detected in the HTM by RT-PCR and Western blot analysis. MIF was also shown by ELISA to be secreted by the HTM cells in culture. The HTM supernatant enhanced IFN-gamma production by TCCs, but not IL-10; and these effects were neutralized by anti-MIF antibodies. Similarly, recombinant MIF enhanced the IFN-gamma production by the TCCs. CONCLUSIONS: MIF is expressed and secreted in the HTM, and MIF has the capacity to enhance T helper 1 cytokines and may play a role as an inflammatory cytokine in the eye.

Monocyte chemotactic protein-1 levels in the vitreous of patients with proliferative vitreoretinopathy.

PURPOSE: To assess the potential role of monocyte chemotactic protein-1 (MCP-1) in the pathogenesis of proliferative vitreoretinopathy (PVR) and to investigate its possible interaction with the macrophage migration inhibitory factor (MIF). METHODS: We assayed MCP-1 and MIF levels in the vitreous samples of 85 consecutive patients with PVR (29 eyes), rhegmatogenous retinal detachment (RRD; 22 eyes), and macular hole or idiopathic epimacular membrane (controls; 34 eyes), by enzyme-linked immunosorbent assay. RESULTS: Vitreous levels of MCP-1 were 1760.7 +/- 471.3 pg/mL (mean +/- SD) in PVR patients, 1200.4 +/- 579.8 pg/mL in RRD patients, and 436.3 +/- 286.1 pg/mL in the controls. Vitreous MCP-1 levels in PVR patients were significantly higher than those in RRD patients and in the controls (P <.0001, respectively). MCP-1 levels in grade C of PVR (1883.7 +/- 479.5 pg/mL) were significantly greater than those in grade D (1437.8 +/- 258.8 pg/mL) (P =.0112). Vitreous concentrations of MCP-1 had no correlation with those of MIF. CONCLUSIONS: The results indicate the possibility that MCP-1 may have a role mainly in the early stage of PVR and that the role of MCP-1 in PVR may differ from that of MIF.