A Case of Herpetic Keratitis after Subconjunctival Triamcinolone Acetonide Injection

Purpose

We report a case of herpetic epithelial keratitis that developed after subconjunctival triamcinolone acetonide injection (STI).

Methods

A 65-year-old female with anterior uveitis and hypotony in her right eye was given a STI (2 mg/0.5 ml). After the injection, she developed redness and an ocular discharge. A clinical examination was performed and real-time polymerase chain reaction (PCR) was used to amplify the viral DNA in a corneal scraping.

Results

Slit-lamp biomicroscopy revealed a severe purulent discharge, conjunctival injection, and a geographic corneal ulcer in the right eye. Herpes simplex virus 1 DNA was identified in the corneal scraping using real-time PCR. Herpetic keratitis was diagnosed and topical acyclovir ointment as well as systemic valacyclovir were started. The inflammation subsided with this medication.

Conclusion

We encountered a case of herpetic epithelial keratitis after a STI.Key words: Herpetic keratitis, Subconjunctival triamcinolone acetonide injection, Anterior uveitis, Polymerase chain reaction, Hypotony     

Comparative Analysis of Polymerase Chain Reaction Assay for Herpes Simplex Virus 1 Detection in Tear

Purpose

To comparatively analyze the methodological efficacy of the polymerase chain reaction (PCR) assay for herpes simplex virus 1 (HSV) detection in tears.

Methods

This retrospective study reviewed the medical records of 115 patients who were clinically diagnosed with herpes keratitis, and their tear samples were collected for HSV detection. PCR positive rates were analyzed for their dependence on the PCR primers used (conventional PCR primer vs. nested PCR primer), the tear collecting method used (micropipetting vs. collection with schirmer strip), the disease manifestation and the patient''s previous medication history.

Results

HSV DNA was detected in 23 out of 115 (20%) tear samples. The PCR positive rate in tear samples did not differ depending on the PCR primer or tear collection method used. Typical epithelial lesions showed a higher positive rate (31.4%) than atypical epithelial lesions (10.9%). The previous history of the antiviral agent seemed to affect the PCR positive rate.

Conclusions

Although the PCR positive rate was not dependent on the tear collection method or primers, HSV detection in tears using PCR was shown to be a supplementary diagnostic test in typical and atypical herpes epithelitis.     

Varicella-Zoster Virus Keratitis with Asymptomatic Conjunctival Viral Shedding in the Contralateral Eye

Purpose

To report a case of varicella-zoster virus (VZV) keratitis with detection of VZV DNA in the tear fluid of not only the symptomatic eye but also the contralateral asymptomatic eye by polymerase chain reaction (PCR).

Methods

This is a case report. A 63-year-old otherwise healthy woman presented with circular corneal ulcer and stromal opacity with infiltration accompanied by mild conjunctival and ciliary injections in the left eye. Bacterial cultures of the corneal scrapings and virus PCR analyses of tear fluid from both eyes were performed.

Results

No pathogen was found by bacterial cultures. PCR was negative for Acanthamoeba, herpes simplex virus and cytomegalovirus, but positive for VZV. VZV DNA was also detected in the unaffected eye. Based on the diagnosis of VZV keratitis, oral valacyclovir and acyclovir eye ointment were administered to the corneal infected eye. The infected eye was healed and VZV DNA turned negative in the tear fluid of the treated eye after 6 months of treatment; however, VZV DNA was still positive in the tear fluid of the contralateral eye.

Conclusions

To our knowledge, this is the first case report of the detection of VZV DNA in the tear fluid of both affected and unaffected eyes in a patient with VZV keratitis. Asymptomatic conjunctival shedding of VZV may continue in the healthy unaffected eye in VZV keratitis patients.Key Words: Varicella-zoster virus, Keratitis, Asymptomatic shedding, Tear fluid, PCR     

Peripheral Ulcerative Keratitis following Laser in situ Keratomileusis

Purpose

We report a case of a patient with a history of glomerulonephropathy, not disclosed prior to laser in situ keratomileusis (LASIK), who developed severe postoperative peripheral ulcerative keratitis (PUK) soon after surgery.

Method

Case report.

Results

Within a week of surgery, the patient, who had no blepharitis or ocular surface disease, also developed diffuse lamellar keratitis (DLK) that was not contiguous with the PUK. Microbiologic evaluation of the flap interface disclosed no organisms, and no epithelial ingrowth was found. Both PUK and DLK resolved with topical and oral steroid therapy, and the patient''s induced refractive error improved over the 12 months following LASIK.

Conclusions

Necrotizing keratitis has been described after LASIK surgery in patients with or without autoimmune disease. However, to our knowledge, there has been no case of PUK following LASIK. As shown by our patient''s clinical course and the typical association of PUK with systemic conditions, patients with a history of atypical postinfectious sequelae may require additional preoperative counseling, vigilant postoperative monitoring, and possibly additional intervention. Because patients do not always divulge medical details, especially if an extraocular site was involved or illness occurred many years prior, this case demonstrates the importance of performing a diligent history that excludes autoimmune disorders or atypical postinfectious sequelae prior to proceeding with keratorefractive intervention.Key Words: Necrotizing keratitis, In situ keratomileusis, Surgically induced keratitis, Peripheral ulcerative keratitis, Autoimmune disease, Postinfectious complication     

Viral causes of unexplained anterior uveitis in Thailand

Aims

To assess the possible role of virus infection in patients with unexplained anterior uveitis (AU).

Methods

Intraocular fluid and plasma samples of 30 HIV-negative AU patients who were unresponsive or poorly responsive to topical steroid therapy were analyzed for nucleic acid of cytomegalovirus (CMV), herpes simplex virus (HSV), and varicella zoster virus (VZV) by real-time polymerase chain reaction (PCR) and for intraocular antibodies against these viruses by Goldmann–Witmer coefficient (GWC) analysis. Of these 30 cases, 21 were tested for rubella virus by GWC analysis, 16 of which also had PCR assessment of aqueous for rubella virus.

Results

Viral uveitis determined by either real-time PCR and/or GWC was documented in 20 out of 30 patients (67%). Of 30 paired samples tested by both methods for HSV, CMV, and VZV, 15 showed positive results (CMV (10), HSV (4), and VZV (1)). Real-time PCR was positive in 8/15 (53%), whereas GWC was positive in 10/15 (67%). Out of 10 CMV-positive patients, four had endotheliitis, two had Posner–Schlossman syndrome, and one Fuchs heterochromic uveitis syndrome (FHUS). Five out of 21 (24%) samples tested by GWC for Rubella virus were positive, three of which exhibited clinical features of FHUS.

Conclusions

Our results indicate that CMV is a major cause of AU in Thailand and show that FHUS can be caused by both CMV and Rubella virus.     

Methicillin-Resistant Staphylococcus aureus Keratitis after Descemet's Stripping Automated Endothelial Keratoplasty

Purpose

We report a case of methicillin-resistant Staphylococcus aureus (MRSA) keratitis after Descemet''s stripping automated endothelial keratoplasty (DSAEK).

Case Report

An 87-year-old woman who had undergone a DSAEK 4 months previously was referred to Tokushima University Hospital with a diagnosis of infectious keratitis after DSAEK. A white abscess and infiltration in the inferior cornea of the right eye were observed. We started an empiric therapy using topical levofloxacin and chloramphenicol on the basis of the microscopic findings of the corneal scraping concurrently with cultivation of the cornea.

Results

A strain of MRSA was isolated from the corneal sample. Although the strain was susceptible to chloramphenicol, it was resistant to quinolone. The keratitis improved rapidly due to empiric therapy, and topical steroids could be resumed 6 days after initiation of the empiric therapy.

Conclusions

To our knowledge, this is the first case of MRSA keratitis, and the second case of bacterial keratitis, after DSAEK. MRSA keratitis can occur following uneventful DSAEK. The empiric therapy on the basis of results from a light microscopic examination of a Gram-stained corneal scraping and restarting topical steroids in the early stages of medication contributed to the good clinical course of this case.Key words: Descemet''s stripping automated endothelial keratoplasty, Methicillin-resistant Staphylococcus aureus, Keratitis     

Tarantula hairs as corneal foreign bodies

Purpose

To report a case of tarantula hairs found in the cornea and discuss treatment.

Case Report

A 16-year-old male presented with a 6-week history of right ocular irritation that began after letting his pet tarantula crawl on his face. Slit-lamp examination of the right eye revealed the presence of approximately 16 dark foreign bodies that had the appearance of small hairs. The foreign bodies were removed from the nasal region of the right cornea using Jewelers forceps, and the patient was prescribed a combination neomycin, polymyxin B, and dexamethasone ointment (Maxitrol^®), given 4 times per day.

Results

The patient presented for follow-up 2 weeks later, with resolution of symptoms.

Conclusion

Effective treatment of keratitis caused by tarantula hairs includes taking a detailed history, conducting a careful slit-lamp examination, removal of any accessible hairs, and initiation of treatment with a topical steroid as determined by the clinical picture.Key Words: Cornea, Keratitis, Foreign body, Tarantula     

Curative effect assessment of bandage contact lens in neurogenic keratitis

AIM:To observe the curative effect of bandage contact lens in neurogenic keratitis.METHODS:Twenty cases of neurogenic keratitis were studied attheDepartment of Ophthalmology, the first Affiliated Hospital of China Medical University, between October 2012 and June 2013. These included 13 males and 7 females, aged from 35 to 88y. Patients were voluntarily divided into an experimental group (lens wearing group, n=10) and control group (drug therapy, n=10). In experimental group patients wore silicone hydrogel bandage soft contact lens. Both groups used the following eyedrops:0.5% levofloxacin TID; 0.5% Sodium carboxymethyl cellulose QID; fibroblast growth factor BID; ganciclovir BID [cases complicated with herpes simplex virus (HSV)]; compound tropicamide BID (cases concurrent hypopyon). The healing time of corneal ulcer and complication rates were observed in the two groups.RESULTS: The healing time of corneal ulcer in the experimental group was 10.80±4.44d versus 46.70±13.88d in the control group (P＜0.05). No complications occurred in the experimental group, except for the lens falling off twice in one case, the patient recovered eight days after rewearing the lens. While in the control group, all cases vascularized, 2 cases were complicated with descemetocele that recovered with amniotic membrane transplantation and 1 case was complicated with corneal perforation that recovered by autologous conjunctival flap covering.CONCLUSION: Bandage contact lens is a safe and effective method of treating neurogenic keratitis and significantly shortened the healing time of corneal ulcer.     

Herpes simplex virus-1 infection or Simian virus 40-mediated immortalization of corneal cells causes permanent translocation of NLRP3 to the nuclei

AIM: To investigate into the potential involvement of pyrin containing 3 gene (NLRP3), a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of corneas against viruses. METHODS: The herpes viral keratitis model was utilized in BALB/c mice with inoculation of herpes simplex virus-1 (HSV-1). Corneal tissues removed during therapy of patients with viral keratitis as well as a Simian vacuolating virus 40 (SV40)-immortalized human corneal epithelial cell line were also examined. Immunohistochemistry was used to detect NLRP3 in these subjects, focusing on their distribution in tissue or cells. Western blot was used to measure the level of NLRP3 and another two related molecules in NLPR3 inflammasome, namely caspase-1 and IL-1β. RESULTS: The NLRP3 activation induced by HSV-1 infection in corneas was accompanied with redistribution of NLRP3 from the cytoplasm to the nucleus in both murine and human corneal epithelial cells. Furthermore, in the SV40-immortalized human corneal epithelial cells, NLRP3 was exclusively located in the nucleus, and treatment of the cells with high concentration of extracellular potassium (known as an inhibitor of NLRP3 activation) effectively drove NLRP3 back to the cytoplasm as reflected by both immunohistochemistry and Western blot. CONCLUSION: It is proposed that herpes virus infection activates and causes redistribution of NLRP3 to nuclei. Whether this NLRP3 translocation occurs with other viral infections and in other cell types merit further study.     

Deep Anterior Lamellar Keratoplasty Using Irradiated Acellular Cornea with Amniotic Membrane Transplantation for Intractable Ocular Surface Diseases

Purpose

To report the clinical outcomes of deep anterior lamellar keratoplasty (DALK) when sterile gamma-irradiated acellular corneal tissues (VisionGraft) are used in combination with amniotic membrane transplantation (AMT) for intractable ocular surface diseases.

Methods

The medical records of fifteen patients who had DALK with AMT were retrospectively reviewed. Indications for surgery included ocular burn, bacterial keratitis, herpes simplex virus keratitis, corneal opacity with Stevens-Johnson syndrome, Mooren''s ulcer, idiopathic myxoid degeneration of corneal stroma, and recurrent band keratopathy. DALK was performed using partial-thickness acellular corneal tissue and a temporary amniotic membrane patch was added at the end of the operation.

Results

All cases that underwent DALK with AMT became epithelialized within 2 postoperative weeks. Twelve patients showed favorable outcomes without graft rejection, corneal opacification, or neovascularization. The other three grafts developed corneal opacification and neovascularization, and required additional penetrating keratoplasty (PK). Unlike the results of previous PKs, there were no graft rejections and the graft clarity was well-maintained in these three cases for at least 8 months after PK.

Conclusions

DALK using sterile acellular corneal tissues in combination with AMT may be a good therapeutic strategy for treating intractable ocular surface diseases because of lowered immune rejection, fibroblast activation, and facilitation of epithelialization. Furthermore, DALK can help stabilize the ocular surface, prolong graft survival, and may allow better outcomes when combined with subsequent PK.     

Corneal Chromoblastomycosis Caused by Fonsecaea pedrosoi

Purpose

To report 2 unusual cases of fungal keratitis due to Fonsecaea pedrosoi.

Methods

Two patients were diagnosed with Fonsecaea pedrosoi keratitis. Their files were reviewed for predisposing factors, clinical characteristics, microbiological study, treatment, and outcome.

Results

Two consecutive patients presented with brownish pigmented corneal ulcers in their eyes after sustaining eye trauma from vegetative matter. In both cases, corneal scrapings were collected for microscopic examination and culture. Dematiaceous hyphae were seen on the smears, and dark pigmented colonies grew on the culture media, identified as F. pedrosoi. Both patients were treated and cured with combined topical antifungal agents and oral itraconazole. The first patient required an amniotic membrane patch, while the second received an intracameral amphotericin B injection.

Conclusions

Pigmented infiltrates can be an important diagnostic clue, but a microscopic evaluation and culture are required to obtain an accurate diagnosis of Fonsecaea keratitis. The prompt diagnosis and combined antifungal treatment can prevent morbidity associated with this fungal infection.Key Words: Fonsecaea pedrosoi, Dematiaceous fungus, Keratitis, Chromoblastomycosis     

Keratitis after Implantation of Intrastromal Corneal Rings with Spontaneous Extrusion of the Segment

Purpose

To report a case of bacterial keratitis in a patient with a history of intrastromal corneal ring segments (INTACS®) implantation to correct keratoconus.

Methods

The patient''s history, clinical presentation, pathological analysis and therapeutic management were reviewed.

Results

A 36-year-old-man was referred to our department due to decreased vision and intense pain in his left eye, 40 days after INTACS® implantation for keratoconus. Slit-lamp examination revealed epithelial defects and stromal infiltrates in the lower channel without evidence of the inferior ring. The anterior chamber also showed a significant fibrin reaction to hypopyon. A low-tension suture was removed at the site of the incision. Microbiological study of the conjunctival swab was positive for Staphylococcus epidermidis, but the corneal culture was sterile. The patient was treated with topical fortified and systemic antibiotics. The infection slowly resolved, leaving opacity at the inferior segment site.

Conclusions

Infectious keratitis following INTACS implantation is an infrequent complication that can have important consequences without suitable and early therapeutic management.Key Words: INTACS®, Intrastromal rings, Keratitis, Segment extrusion     

Double-biguanide therapy for resistant acanthamoeba keratitis

Aims

To report the clinical and diagnostic findings of a patient with Acanthamoeba keratitis resistant to both polyhexamethylene biguanide (PHMB)-hexamidine and chlorhexidine-hexamidine treatment.

Methods

Slit-lamp biomicroscopy, corneal cell scraping and histopathology were performed on a 39-year-old woman presenting with corneal ulcer in her left eye.

Results

The patient was successfully treated with PHMB-chlorhexidine association therapy. Subsequent perforating keratoplasty remained clear at the last follow-up visit after 7 months and increased visual acuity to 20/20 with correction.

Conclusions

This case emphasizes the proteiform aspects of Acanthamoeba drug resistance, and suggests that PHMB-chlorhexidine association might represent an additional option for cases resistant to standard therapy.Key Words: Acanthamoeba keratitis, Biguanide, Diamidine     

Peripheral ulcerative keratitis and necrotizing scleritis initiated by trauma in the setting of mixed cryoglobulinemia

Purpose

To report a case of peripheral ulcerative keratitis and necrotizing scleritis precipitated by trauma in a patient with mixed cryoglobulinemia due to hepatitis C viral infection.

Methods

Case report and literature review.

Results

A 62-year-old man with a history of mixed cryoglobulinemia developed an episode of necrotizing scleritis and peripheral ulcerative keratitis one month after repair of a traumatic scleral defect with patch grafting. This episode resolved following treatment with high-dose corticosteroids and the patient underwent successful repeat patch grafting along with a free conjunctival autograft. This is the second reported case of necrotizing scleritis and peripheral ulcerative keratitis associated with mixed cryoglobulinemia.

Conclusion

Ophthalmologists should be aware of the association between mixed cryoglobulinemia and necrotizing scleritis/peripheral ulcerative keratitis. Patients with this condition experiencing ocular trauma or undergoing ocular surgery should be monitored closely.Key words: Scleritis, Peripheral ulcerative keratitis, PUK, Mixed cryoglobulinemia, Hepatitis C     

Improvement in corneal scarring following bacterial keratitis

Aim

Bacterial keratitis results in corneal scarring and subsequent visual impairment. The long-term evolution of corneal scars has not been well described. In this case series, we identified patients who had improvement in corneal scarring and visual acuity from a clinical trial for bacterial keratitis.

Methods

We searched the records of the Steroids for Corneal Ulcers Trial (SCUT) for patients who had improvement in vision between the 3-month and 12-month visits and reviewed their clinical photographs.

Results

Of the 500 patients enrolled in SCUT, five patients with large central corneal scars due to bacterial keratitis are presented. All experienced improvement in rigid contact lens–corrected visual acuity from months 3 to 12. All patients also had marked improvement in corneal opacity during the same time period. None of the patients opted to have penetrating keratoplasty.

Conclusions

Corneal scars may continue to improve even many months after a bacterial corneal ulcer has healed. The corneal remodeling can be accompanied by considerable improvement in visual acuity, such that corneal transplantation may not be necessary.     

Acute Retinal Necrosis in Childhood

Background

Acute retinal necrosis (ARN) is a viral syndrome consisting of uveitis/vitritis, occlusive vasculitis and peripheral necrosis. Few incidents are reported in children. The etiology is reactivated herpes simplex virus (HSV) or varicella-zoster virus (VZV). Treatment with acyclovir is often used. The administration of oral glucocorticosteroids is of unproven benefit. Prognosis is variable but poor.

Methods

Three weeks after contracting mild chickenpox, a healthy 4-year-old girl developed blurred vision in her right eye. Severely reduced visual acuity was noted, together with anterior uveitis, ‘mutton-fat’ precipitates and vitral flare. Retinal vasculitis with necrosis was present. Serology for toxoplasma, cytomegalovirus and HIV was negative, while HSV and VZV IgG antibodies were positive. She was treated with 30 mg/kg of intravenous methylprednisolone (3 days), 30 mg of oral prednisone (3 days), and tapering for 8 weeks. Intravenous acyclovir was given for 10 days, followed by oral acyclovir for 4 months. Aspirin (100 mg/day) was given for 4 months.

Results

At 12 months, the girl felt good. Her right eye acuity was 6/9, with an intraocular pressure of 17 mm Hg. The peripheral retina showed scarring but no detachment.

Conclusions

This is the first report of a once-daily high-dose methylprednisolone pulse therapy in one of the youngest known ARN cases. Pulsed steroid therapy was based on its known effectiveness in vasculitis, which is the main pathophysiology in ARN. There was no evidence of steroid-related viral over-replication. Our case achieved an excellent clinical and ophthalmic recovery in spite of the poor prognosis. The positive result of this case report provides a basis for further evaluation of high-dose steroid pulse therapy in ARN.Key words: Retinal necrosis, Children, Methylprednisolone     

Fungal Keratitis – Improving Diagnostics by Confocal Microscopy

Purpose

Introducing a simple image grading system to support the interpretation of in vivo confocal microscopy (IVCM) images in filamentous fungal keratitis.

Setting

Clinical and confocal studies took place at the Department of Ophthalmology, Aarhus University Hospital, Denmark. Histopathological analysis was performed at the Eye Pathology Institute, Department of Neuroscience and Pharmacology, University of Copenhagen, Denmark.

Methods

A recent series of consecutive patients with filamentous fungal keratitis is presented to demonstrate the results from in-house IVCM. Based upon our experience with IVCM and previously published images, we composed a grading system for interpreting IVCM images of filamentous fungal keratitis.

Results

A recent case series of filamentous fungal keratitis from 2011 to 2012 was examined. There were 3 male and 3 female patients. Mean age was 44.5 years (range 12–69), 6 out of 17 (35%) cultures were positive and a total of 6/7 (86%) IVCM scans were positive. Three different categories of IVCM results for the grading of diagnostic certainty were formed.

Conclusion

IVCM is a valuable tool for diagnosing filamentous fungal keratitis. In order to improve the reliability of IVCM, we suggest implementing a simple and clinically applicable grading system for aiding the interpretation of IVCM images of filamentous fungal keratitis.Key words: In vivo confocal microscopy, Fungal keratitis, Diagnostics     

Ultraviolet-a light and riboflavin therapy for acanthamoeba keratitis: a case report

Purpose

To report ultraviolet-A (UV-A) light treatment in a patient with Acanthamoeba keratitis (AK).

Methods

Interventional case report. A standard protocol for ultraviolet corneal therapy, with a power emission of 3 mW/cm² and a wavelength of 370 nm, was used. The protocol included an 8-nm bandwidth at a 54-mm distance measured with a collimation system of diodes as well as a protective shield of riboflavin in a case of documented AK.

Results

A 54-year-old female patient with AK, showing no therapeutic response to a wide variety of topical antimicrobial agents and with a visual acuity of 20/400, was treated with UV-A therapy. The patient displayed a favourable response in the first 24 h after treatment, with improvement of symptoms, visual acuity (to 20/200) and biomicroscopy cornea with haze degree I. By the third week post-treatment, the patient was symptom-free. Her visual acuity was 20/30, and the affected cornea was clear. Five months after treatment, there had been no recurrence, and her vision was 20/20.

Conclusions

Treatment with UV-A light was an effective therapy in this case of AK.Key words: Acanthamoeba keratitis, Ultraviolet light, Riboflavin     

Persisting epithelial herpes simplex keratitis while on cyclosporin-A ointment

Background: This is the first report of cyclosporin-A ointment (CsA) adversely affecting epithelial herpes simplex virus keratitis in a corneal graft.
Methods: The details of this case were obtained from the patient's record at The Wilmer Ophthalmological Institute.
Results: Ceasing CsA ointment resulted in resolution of the epithelial HSV keratitis.
Conclusion: Immunosuppression caused by CsA ointment resulted in persisting epithelial HSV keratitis despite adequate topical treatment.     

Prospective comparison between conventional microbial work-up vs PCR in the diagnosis of fungal keratitis

Purpose

The PCR was compared with routine microbial studies for the detection of fungal pathogens in clinically suspected fungal keratitis.

Methods

A prospective nonrandomized study was undertaken at a tertiary eye care centre to evaluate 30 eyes of 30 patients with presumed fungal keratitis, both fresh and treated. Corneal scrapings were performed on each patient. The specimens were analysed by a semi-nested PCR assay using fungal-specific primers. PCR products were cloned and sequenced for identification, and compared with a conventional microbial work-up (smear and culture).

Results

Of the 30 samples, the PCR showed positivity in 93.3%, culture in 40%, and potassium hydroxide in 20%. Of the 28 PCR-positive cases, 12 were culture-positive and 16 were culture-negative. Two samples were both PCR and culture test negative. Culture-negative samples were PCR-positive in 16 of 18 (88.9%) cases. The PCR did not yield any false-negative findings in a culture-positive specimen. Both common and uncommon aetiologic fungi have been identified by DNA sequencing analysis.

Conclusion

The PCR was able to detect fungal DNA in a high proportion of culture-negative cases. Technical considerations of the PCR process include extraction of artifacts and amplification of non-pathogenic DNA. Nonetheless, our findings suggest that the PCR can be a useful adjunct to smear and culture in the rapid diagnosis of fungal keratitis, particularly in cases of failed detection from routine procedures.