Biological characteristics and clinical management of uveal and conjunctival melanoma

Uveal and conjunctival melanomas are relatively rare tumors;nonetheless,they pose a significant risk of mortality for a large number of affected individuals.The pathogenesis of melanoma at different sites is very similar,however,the prognosis for patients with ocular melanoma remains unfavourable,primarily due to its distinctive genetic profile and tumor microenvironment.Regardless of considerable advances in understanding the genetic characteristics and biological behaviour,the treatment of uveal and conjunctival melanoma remains a formidable challenge.To enhance the prospect of success,collaborative efforts involving medical professionals and researchers in thefields of ocular biology and oncology are essential.Current data show a lack of well-designed randomized clinical trials and limited benefits in current forms of treatment for these tumors.Despite advancements in the development of effective melanoma therapeutic strategies,all current treatments for uveal melanoma(UM)and conjunctival melanoma(CoM)remain unsatisfactory,resulting in a poor long-term prognosis.Ongoing trials offer hope for positive outcomes in advanced and metastatic tumors.A more comprehensive understanding of the genetic and molecular abnormalities involved in the development and progression of ocular melanomas opens the way for the development of personalized therapy,with various potential therapeutic targets currently under consideration.Increased comprehension of the molecular pathogenesis of UM and CoM and their specificities may aid in the development of new and more effective systemic therapeutic agents,with the hope of improving the prognosis for patients with metastatic disease.

Preliminary study on the efficacy of lacrimal duct reconstruction with pedicled conjunctival flap in the treatment of severe lacrimal canalicular obstruction with conjunctivochalasis

AIM:To determine the efficacy and safety of pedicled conjunctival lacrimal duct reconstruction in the treatment of severe obstruction of superior and inferior lacrimal canaliculi with conjunctivochalasis.METHODS:This study was per formed as a retrospective analysis of patients who received conjunctival dacryocystorhinostomy with pedicled conjunctival flap reconstruction combined with tube intubation due to severe superior and inferior lacrimal canalicular obstruction with conjunctivochalasis from January 2019 to October 2019.The clinical data included the degree of preoperative epiphora and postoperative relief,preoperative examination of lacrimal duct computed tomography and ultrasound biomicroscopy,postoperative evaluation of lacrimal duct function by chloramphenicol taste and fluorescein dye disappearance test,etc.Syringing was carried out to determine the reconstruction and patency of the lacrimal duct.RESULTS:All 9 patients(9 eyes)had severe canalicular obstruction with conjunctivochalasis.The patients included 4 males and 5 females aged between 47–65y with an average age of 52.2±6.7y.At 3mo follow-up,the tube was removed and the patients were followed for a further 3mo.After tube removal,6 patients showed no epiphora.These patients also had positive chloramphenicol tastes and normal fluorescein dye disappearance test results.Two patientshad epiphora.Also,syringing showed partial patency of the reconstructed lacrimal duct.One patient had no improvement in epiphora with negative chloramphenicol taste and fluorescein dye disappearance test results and obstruction of the reconstructed lacrimal duct.The total effective rate of the operation was 8/9,with no serious complications.CONCLUSION:Pedicled conjunctival lacrimal duct reconstruction conjunctival dacryocystorhinostomy is safe and effective for superior and inferior canalicular obstruction with conjunctivochalasis.

Clinical outcome of combined conjunctival autograft transplantation and amniotic membrane transplantation in pterygium surgery

AIM： To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques： combined conjunctival autograft and overlay amniotic membrane transplantation （CAT with AMT）, conjunctival autograft transplantation （CAT） alone and amniotic membrane transplantation （AMT） alone. METHODS： In this retrospective study, 142 eyes of 142 pterygium patients （104 primary, 38 recurrent）who underwent CAT （group A）, AMT （group B） or CAT with AMT （group C） respectively following surgical excision were reviewed and compared based on the recurrences and post-operative complications. RESULTS： The number of recurrence post-surgery were 17 （9 from primary, 8 from recurrent; the same description below）, 18 （10, 8） and 2 （1, 1） in groups A, B, and C respectively; dry eyes were 22 （16, 6）, 27 （18, 9） and 7 （3, 4）; conjunctival inflammations were 30 （17, 13）, 27 （16, 11） and 11 （6, 5）. Patients in group C （either pdmary or recurrent or both） mainly showed significantly better results than those in group A or B （P〈0.05） regarding above-mentioned clinical effects. CONCLUSION： Combined CAT and overly AMT have significantly lower rates of recurrence and postoperative complications for primary and recurrent pterygium surgery than CAT or AMT alone.

Clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis

AIM: To study the clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis. METHODS:A retrospective study was done to 10 patients (10 eyes) who had accepted removal of the necrotic corneal tissue combined with conjunctival flap covering surgery for fungal keratitis,the diagnosis by corneal scraping and smear examination or confocal microscopy check hyphae. Local and systemic antifungal therapy more than one week for all patients, corneal ulcer enlarge or no shrink. Slit lamp microscope examination the diameter of corneal ulcer about 2mm-4mm. Anterior segment optical coherence tomography (AS-OCT)examine the depth of corneal ulcer between 1/3-1/2, infiltrate corneal stroma about 20um-80um,the diameter of corneal ulcer about 3mm-6mm.Type-B ultrasonic exclusion endophthalmitis. Complete removal lesions until transparent of stoma, make conjunctival flap equal or greater than ulcer 1mm nearby conjunctiva. Continued antifungal therapy. The vision, fungal recurrence, conjunctival flap rollback or desquamate were analysed. ' RESULTS:Ten patients had success done this surgery, the corneal ulcer was not enlarge and healing afteroperation. 7 cases were bridging conjunctival flap and 3cases were single conjunctival flap. Preoperation vision above 0.1 had 8 cases,7 cases had vision above 0.1 one week after surgery, while 1 cases vision droped from 0.3 to 0.05.There was not recurrent for fungal,2 cases conjunctival flap rollback:1 case was bridging and 1case was single flap, no conjunctival flap desquamate. CONCLUSION: It is safe and effective to perform removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis which werenot sensitive or aggravate for antifungal drugs.

Treatment of intractable orbital implant exposure with a large conjunctival defect by secondary insertion of the implant after preceding dermis fat graft

AIM: To report a procedure and results of a two-stage operation to manage intractable extensive orbital implant exposure with a large conjunctival defect which was difficult to treat with dermis fat grafts due to repeated graft necrosis. METHODS: A retrospective chart review of four patients who had extensive orbital implant exposures with large conjunctival defects and had past histories of repeated autologous or preserved dermis graft failures was done. As a first-stage operation, the problematic pre-existing orbital implants were removed and autologous dermis fat grafts alone were performed on the defect area. Four months later, new orbital implants were secondarily inserted after confirmation of graft survival. The size of the conjunctival defects and state of the extraocular muscles were checked preoperatively. Success of the operations and complications were investigated.RESULTS: The mean size of the conjuctival defects was 17.3mm×16.0mm, and the mean time from the initial diagnosis of orbital implant exposure to implant removal and autologous dermis fat graft was 20.8 months. After implant removal and autologous dermis fat graft, no graft necrosis was observed in any patients. Also, implant exposure or fornix shortening was not observed in any patients after new orbital implant insertion. CONCLUSION: The secondary insertion of a new orbital implant after pre-existing implant removal and preceding dermis fat graft is thought to be an another selective management of intractable orbital implant exposure in which dermis fat grafts persistently fail.

Effect of long-term topical latanoprost medication on conjunctival thickness in patients with glaucoma

AIM： To investigate the effect of long-term use of topically administered latanoprost on conjunctival thickness（CT） and conjunctival epithelium thickness（CET） in the patients with glaucoma. METHODS： A series of 106 glaucomatous patients were included. Of the 106 eyes, 55 eyes were treated with latanoprost eye drops once a day（latanoprost group）, while 51 eyes were treated with carteolol hydrochloride eye drops（carteolol group）. All the included patients completed a 2-year follow-up. CT and CET were measured with optical coherence tomography（OCT） in all patients at presentation and at 2-year visit, respectively. Statistical analysis was then performed to compare the change in CT and CET. RESULTS： At presentation, there was no difference in CET（t=0.400, P=0.689） or CT（t=1.14, P=0.259） between the two groups. No significant difference was found in CET（61.65±5.35 μm at baseline, 60.36±6.36 μm at 2-year follow-up, respectively; t=1.977, P=0.0531）, while there was a significant decrease in CT from 201.45±14.99 μm at baseline to 167.81±14.57 μm at 2-year visit（t=14.1407, P〈0.001） in the latanoprost group. At 2-year follow-up, no statistically difference was found in CET（62.24±5.27 μm; t=1.086, P=0.282） or CT（201.23±12.45 μm; t=1.44, P=0.154） compared to it at baseline（CET： 61.23±5.42 μm; CT： 198.76±13.68 μm, respectively） in the carteolol group. CONCLUSION： A significant decrease in conjunctival thickness is found in glaucoma patients treated with long-term topical latanoprost; its potential effect on the outcome of filtration surgery should be considered.

In vitro adherence of conjunctival bacteria to different oculoplastic materials

AIM: To investigate the resistance to bacterial adhesion of materials used in oculoplastic surgery, particularly materials used in the manufacture of orbital implants.METHODS: Seven organisms of conjunctival flora(two strains of Staphylococcus epidermidis and one strain each of Staphylococcus aureus, Staphylococcus hominis, Corynebacterium amycolatum, Acinetobacter calcoaceticus, and Serratia marcescens) were selected. A lactic acid bacterium(Lactobacillus rhamnosus) was also included as positive control because of its well-known adhesion ability. Eight materials used to make oculoplastic prostheses were selected(glass, steel, polytetrafluoroethylene, polymethylmethacrylate, silicone from orbital implants, commercial silicone, porous polyethylene, and semismooth polyethylene). Materials surfaces and biofilms developed by strains were observed by scanning electron microscopy. Kinetics of growth and adhesion of bacterial strains were determined by spectrophotometry. Each strain was incubated in contact with plates of the different materials. After growth, attached bacteria were re-suspended and colony-forming units(CFUs) were counted. The number of CFUs per square millimetre of material was statistically analyzed.RESULTS: A mature biofilm was observed in studied strains except Staphylococcus hominis, which simply produced a microcolony. Materials showed a smooth surface on the microbial scale, although steel exhibited 1.0-μm-diameter grooves. Most organisms showed significant differences in adhesion according to the material. There were also significant differences in thetotal number of CFUs per square millimetre from each material(P=0.044). CFU counts were significantly higher in porous polyethylene than in silicone from orbital implants(P=0.038).CONCLUSION: Silicone orbital implants can resist microbial colonization better than porous polyethylene implants.

Sulforaphane modulates TGFβ2-induced conjunctival fibroblasts activation and fibrosis by inhibiting PI3K/Akt signaling

AIM:To examine the effects of sulforaphane on fibrotic changes of transforming growth factor(TGFβ2)induced human conjunctival fibroblast(HCon Fs).METHODS:HCon Fs were cultured and divided into control,TGFβ2(1 ng/m L),sulforaphane and TGFβ2+sulforaphane groups.Cell viability and apoptosis were detected using the MTT and Apo Tox-Glo Triplex assay.Cell migration was detected using scratch and Transwell assay.Real-time quantitative PCR method was used to evaluate m RNA expression of TGFβ2,matrix metalloproteinase-2(MMP2),myosin light chain kinase(MYLK),integrinαV,integrinα5,fibronectin 1 andα-smooth muscle actin(α-SMA).The protein expression ofα-SMA,p-PI3 K,PI3 K,p-Akt,and Akt were detected by Western blot.RESULTS:The proliferation of HCon Fs was significantly(P<0.05)suppressed by sulforaphane compared to control cells with the increase of the concentration and treatment time.Cell proliferation after 48 h incubation was significantly reduced with 100μmol/L sulforaphane treatment by 17.53%(P<0.05).The Transwell assay showed sulforaphane decreased cell migration by 18.73%compared with TGFβ2-induced HCon F(P<0.05).TGFβ2-induced the increasing expression of fibronectin,type I collagen andα-SMA,and the phosphorylation of PI3 K and Akt were all significantly suppressed by sulforaphane pretreatment.CONCLUSION:Sulforaphane inhibits proliferation,migration,and synthesis of the extracellular matrix in HCon Fs,and inhibiting the PI3 K/Akt signaling pathway.Sulforaphane could be a potential therapeutic drug for prevention of scar formation in filtering bleb after trabeculectomy.

Comparative clinical study of conjunctival toxicities of newer generation fluoroquinolones without the influence of preservatives

AIMTo compare the conjunctival epithelial toxicities of three newer-generation fluoroquinolones without preservatives.METHODSIn a prospective, randomized, double blind comparative study, 47 eyes of 47 patients with a primary pterygium were enrolled, and divided randomly into three groups (levofloxacin 0.5%, gatifloxacin 0.3%, and moxifloxacin 0.5%). After pterygium surgery with the same conjunctival autograft technique, each patient maintained a regimen with a randomly assigned fluoroquinolone eye drop. Patients were examined every other day after surgery until the epithelium had completely healed. Photos were taken and used to measure the area of residual epithelial defects. Conjunctival healing time and speed (initial defect area/healing time (mm2/d) compared in each group using Kruskal-Wallis tests.RESULTSThere were no significant differences in mean age, gender, and conjunctival defect size of the donor site between these groups. However, the mean of conjunctival healing time and speed were statistically different in each group. The mean of conjunctival epithelial healing time was 8.93±2.69d (levofloxacin group), 10.31±2.96d (gatifloxacin group), and 13.50±4.10d (moxifloxacin group), P=0.006. The mean conjuctival epithelial healing speed was 6.18±1.39 mm2/d (levofloxacin group), 5.52±1.68 mm2/d (gatifloxacin group), and 4.40±1.30 mm2/d (moxifloxacin group), P=0. 003.CONCLUSIONWithout the influence of preservatives, levofloxacin and gatifloxacin might be less toxic to the regeneration of conjunctival epithelial cells and cause a faster conjunctival wound healing relative to moxifloxacin.

Two-step strategy—conjunctival flap covering surgery combined with secondary deep anterior lamellar keratoplasty for the treatment of high-risk fungal keratitis

AIM:To investigate whether the two-step strategy[conjunctival flap covering surgery(CFCS)combined with secondary deep anterior lamellar keratoplasty(DALK)]is effective for patients with high-risk fungal keratitis(FK).METHODS:In this noncomparative,retrospective case series,10 subjects(6 males,4 females)with a mean age of 56.5±7.1(range 47-72)y with high-risk FK undergone the two-step strategy were included.Reported outcome measures were healing of the corneal ulcer,recurrence of FK,reject reaction,improvement in best corrected visual acuity(BCVA)and relevant complications.RESULTS:The average diameter of corneal infiltrates was 7.50±0.39 mm,ranging from 6.94 to 8.13 mm.The mean depth of corneal infiltrates was 422.4±77.1μm,ranging from 350 to 535μm.The mean corneal thickness was 597.4±117.3μm,ranging from 458 to 851μm.Hypopyon and endothelial plaques were presented in all patients.The period between the two steps was 3.65±0.9(ranging from 3 to 5)mo.The graft diameter was 7.75±0.39 mm.At the last follow-up(average 9.25±3.39,ranging from 5.5 to 17mo),no fungal recurrence or graft rejection appeared,and all patients showed improvement of BCVA.One patient suffered from liver function impairment due to oral voriconazole for 4wk and recovered spontaneously after 1wk of drug withdrawal.CONCLUSION:The two-step strategy is safe and effective in the treatment of high-risk FK by transforming intentional therapeutic penetrating keratoplasty during acute infection to later optical DALK.It is a practical strategy,especially in areas lacking fresh donor corneas and eye bank services.

Ocular manifestations and SARS-CoV-2 detection in tears and conjunctival scrape from non-severe COVID-19 patients

AIM:To explore the ocular features of corona virus disease(COVID)-19 and severe acute respiratory syndrome coronavirus(SARS-CoV)-2 detection in tears and conjunctival scrapes in non-severe COVID-19 patients.METHODS:This is a multicenter observational clinical study with no intervention conducted from Jan 25th to March 1st,2020.Clinical data and samples of tears and conjunctival scraping were collected in consecutive laboratory-confirmed,non-severe COVID-19 patients from three hospitals.COVID-19 virus was analyzed by real-time reverse transcriptase polymerase chain reaction(RT-PCR)kits.RESULTS:Totally 255 laboratory-confirmed,non-severe COVID-19 patients were recruited for ocular manifestation investigation.Of them,54.9%were females,with a mean age of 49.4 y.None of the patients has evidence of uveitis;11 patients(4.3%)complained of mild asthenopia;2(0.8%)had mild conjunctival congestion and serous secretion.Twenty-five of them had performed tears and conjunctival scrape for COVID-19 virus detection,with 4 yield possible positive results in the nucleoprotein gene.One of them were asymptomatic with normal chest CT and positive pharyngeal swab result.CONCLUSION:Ocular manifestations are neither common nor specific in non-severe COVID-19 patients.Meanwhile,COVID-19 virus nucleotides can be detected in the tears and conjunctival scrape samples,warranting further research on the transmissibility by the ocular route.

Chronic ocular GVHD:limbal and conjunctival stem cell allografts from the same hematopoietic stem cell donor

Dear Editor,I am Dr.Elias F Jarade from the Beirut Eye and ENT Specialist Hospital, Beirut, Lebanon. I write to describe a novel surgical technique in the management of chronic ocular graft-versus-host disease（GVHD）.

Conjunctival flap with auricular cartilage grafting: a modified Hughes procedure for large full thickness upper and lower eyelid defect reconstruction

AIM:To investigate the suitability of a modified Hughes procedure,which consists of conjunctival flap with auricular cartilage grafting in reconstructing large full thickness upper and lower eyelid defect.METHODS:Patients with full thickness eyelid carcinoma involving more than 50%margin length who underwent surgical resection were retrospectively reviewed in the study.The defects were reconstructed using conjunctival flap with auricular cartilage grafting,covered with myocutaneous flap above.Followed-up time ranged from 12 to 24 mo.Outcomes were classified as“good”,“fair”,and“poor”by evaluating the margin appearance,eyelid appearance,and complications.RESULTS:A total of 42 patients were enrolled in the study(26 males,16 females,mean age,68.6±7.7 y,range:53 to 82 y).The mean defect widths measured 23.2±2.9 mm(range,17 to 28 mm).The mean posterior lamellar defect height was 5.5±1.3 mm(4 to 8 mm).Thirty-seven patients had a“good”outcome(88.1%),5 patients had a“fair”outcome(11.9%),and no one had a“poor”outcome.CONCLUSION:Conjunctival flap with auricular cartilage grafting and myocutaneous flap grafting is an effective procedure in reconstructing large full thickness upper and lower eyelid defect.It can not only achieve satisfied reconstruction,but also preserve intact tarsal plate of the opposite eyelid,avoiding retraction or entropion.

Bulbar conjunctival vascular lesion combined with spontaneous retrobulbar hematoma:A case report

BACKGROUND Orbital hemorrhage can be classified as traumatic or spontaneous depending on its cause.Spontaneous orbital hemorrhage refers to an internal orbital hemorrhage without apparent cause.Therefore,we aimed to describe a case of an orbital hematoma after a severe cough the night before due to inhalation of cooking oil fumes.CASE SUMMARY A 46-year-old woman was referred to our hospital with a complaint of exophthalmos accompanied with blurred vision,pain,binocular diplopia,and dizziness lasting for 5 h noted on waking in the morning.She also experienced nausea and vomiting due to high pressure of orbit and dizziness.Based on the auxiliary examination and her medical history,the patient was finally diagnosed with bulbar conjunctival vascular lesion combined with spontaneous retrobulbar hematoma.The patient was administered tobramycin and dexamethasone eye ointment,and applied pressure dressing on the left eye to stop the bleeding.Simultaneously,we administered intravenous etamsylate,oral Yunnan Baiyao capsule,intravenous mannitol to reduce orbital pressure,and intravenous dexamethasone injection at 10 mg/dL combined with neurotrophic therapy to reduce tissue edema.Among them,the Yunnan Baiyao capsule is a traditional Chinese herbal medicine to remove stasis and stop bleeding;thus,it promotes blood circulation and relieves pain resulting in reduced edema of the lesion site.The symptoms did not improve significantly during the first 2 d of treatment.We speculate that high orbital pressure and binocular diplopia induced frequent nausea and vomiting in the patient,causing increased pressure on the superior vena cava and leading to repeated orbital bleeding.After the second day,the symptoms started gradually improving.CONCLUSION This case further emphasizes the importance of comprehensive,detailed medical history and careful ophthalmic examination of the patient.

Ocular leech infestation initially misdiagnosed as conjunctival pigmented nevus

Dear sir,Iam Dr.Wen-Wei Li,from the Department of Ophthalmology,Zhejiang Provincial People’s Hospital,Hangzhou,China.I write to present a case of ocular leech infestation initially misdiagnosed as conjunctival pigmented nevus.Although human interaction with leeches is not uncommon

Efficacy of autologous conjunctival flap on repairing the late-onset filtering bleb leakage

AIM： To evaluate the effectiveness of autologous conjunctival flap surgery for repairing the late-onset filtering bleb leakage after trabeculectomy.METHODS： This study retrospectively reviewed 106 eyes from 106 patients with late-onset filtering bleb leakage after trabeculectomy who received autologous conjunctival flap surgery at the Zhongshan Ophthalmic Centre from 2005 to 2015. The basic information was recorded, and the interval time between trabeculectomy and autologous conjunctival flap surgery as well as related risk factors, intraocular pressure（IOP）, anterior chamber depth（ACD） and best corrected visual acuity（BCVA） were analysed. Moreover, 41 patients who completed the 1-year follow up were analysed to determine the IOP and BCVA changes and long-term success rates.RESULTS： The 50 male and 56 female subjects（average age 39.13±17.96 y） included 47（44.34%） and 33（31.13%） cases of primary open angle and primary angle-closure glaucoma. The mean interval between trabeculectomy and repair surgery was 60.60±56.92（3-264）mo. The mean mitomycin（MMC） concentration during trabeculectomy was 0.27±0.04（0.12-0.4） mg/m L in the fornix-based conjunctival flap group（68 patients） and 0.28±0.04（0.20-0.33） mg/m L in the limbal-based conjunctival flap group（11 patients）. After bleb leakage, the patients＇ vision remained stable while the IOP decreased from 10.25±4.76（3-20.86） to 9.44±4.33（2-21） mm Hg（P〈0.01）. In the 41 analysed patients, the IOP was controlled at 15.68±5.11（7-40） mm Hg in the 1^（st） year after autologous conjunctival flap surgery and recurrence was not observed, for a long-term success rate of 100%. CONCLUSION： Autologous conjunctival flap repairing surgery is an effective technique for sealing filtering bleb leakages and controlling IOP postoperatively.

Conjunctival cytological examination,bacteriological culture,and antimicrobial resistance profiles of healthy Mediterranean buffaloes(Bubalus bubalis) from Southern Italy

Objective: To assess normal conjunctival cytological and bacteriological/fungal flora features in the Mediterranean buffalo(Bubalus bubalis).Methods: Swabs were taken from the inferior conjunctival sac of both eyes of 57 healthy female buffaloes aged 24–36 months, with no evidence of ocular disease, farmed in Campania region(Southern Italy), for microbiological analysis. Conjunctival eye specimens of both eyes were subsequently obtained by a cyto-brush, for cytological analysis.The antimicrobial susceptibility of bacterial isolates was also determined using the diskdiffusion method on Mueller Hinton agar plates.Results: Cytological examination of conjunctival swab specimens(114 eyes) revealed epithelial cells(basal, intermediate, columnar and super ficial) in all samples, whereas neutrophils, lymphocytes and plasma cells were present in 70%, 10% and 2% of samples,respectively. Microorganisms, for a total of 261 aerobic bacteria and 6 fungi, were isolated from 112/114 conjunctival samples [98.25%; 95% con fidence interval(CI): 93.18–99.70]. Only two conjunctival swabs did not yield bacteria and/or fungi(2/114, 1.75%;95% CI: 0.30–6.82). Gram-positive aerobes were most commonly cultured(181/261,69.35%; 95% CI: 63.31–74.81), with Enterococcus faecium and Staphylococcus lentus predominating. Escherichia coli was the most frequently isolated as Gram-negative bacteria(80/261, 30.65%; 95% CI: 25.19–36.69). The antimicrobial resistance patterns of the isolated bacteria showed amoxycillin/clavulanic acid and cephalothin as the least sensitive antibiotics for both Gram-positive and Gram-negative bacteria.Conclusions: These results provided first information on normal conjunctival ocular micro flora and cytological features in Mediterranean buffalo.

Conjunctival flora in anophthalmic patients: microbiological spectrum and antibiotic sensitivity

AIM: To identify the spectrum and susceptibility pattern of isolated microorganisms from conjunctival flora of anophthalmic patients.METHODS: A cross-sectional clinical study including 60 patients with unilateral anophthalmia. Patients with use of antibiotic drops in their socket during the last month were also included. From each patient, three microbiological samples were taken from the lower conjunctival sac(healthy eye, pre-prosthesis, and retro-prosthesis space of socket). The 180 samples obtained were cultured. Isolates were identified and their antibiotic sensitivities were determined.RESULTS: A total of 251 isolates were recovered(62 isolates from healthy eye, 93 from pre-prosthesis, and 96 from retro-prosthesis space). The most common organism was Staphylococcus epidermidis, in both healthy eyes(64.5%) and sockets(45.5%). Altogether, coagulasepositive Staphylococci, Streptococci, and Gram-negative bacteria accounted for less than 15% of isolates in healthy eyes and more than 35% in sockets. Regarding the antibiotic sensitivities, there were no significant differences between isolates from sockets and healthy eyes. Nine patients recognized the use of self-prescribed antibiotic drops in their socket. In the healthy eyes of these subjects, Gram-positive microorganisms showed significantly greater resistance to aminoglycosides and tetracycline.CONCLUSION: Sockets of anophthalmic patients show a greater number of pathogens compared to healthy eyes.The use of antibiotic drops in the socket promotes a resistant flora not only in the socket but also in the healthy eye. Quinolones and macrolides may be better therapeutic options than aminoglycosides for treating conjunctivitis of anophthalmic sockets, since these antibiotics are less active against Staphylococcus epidermidis.

The protective effect of zeaxanthin on human limbal and conjunctival epithelial cells against UV-induced cell death and oxidative stress

AIM: To explore the protective effect of zeaxanthin on human limbal and conjunctival epithelial cells against UVradiation and excessive oxidative stress.METHODS: Human limbal and conjunctival epithelial cells were isolated from cadaver and cultured in vitro. They were challenged with UVB radiation and H2 O2 with and without zeaxanthin pretreatment. Cell viability, p38 and c-JUN NH(2)-terminal kinase(JNK) phosphorylation, IL-6, IL-8 and MCP-1 secretion and malondialdehyde(MDA) content were measured.RESULTS: Zeaxanthin had no measurable cytotoxicity on limbal or conjunctival epithelial cells when used at concentrations of 5 μg/mL and below. At 30 mJ/cm2 UVB, the pretreatment of zeaxanthin increased the percentage of live cells from 50% to 69%(P=0.01) and from 66% to 75%(P=0.05) for limbal and conjunctival epithelial cells, respectively. The concentrations of IL-6, IL-8 and MCP-1 in the culture medium reduced to 66%(for IL-6 and MCP-1)and 56%(for IL-8) of the levels without zeaxanthin. This was accompanied by reduced p38 and JNK protein phosphorylation. Pretreatment of zeaxanthin also reduced intracellular MDA content caused by H2 O2 stimulation from 0.86 μmol/L to 0.52 μmol/L(P=0.02) in limbal epithelial cells and from 0.96 μmol/L to 0.56 μmol/L in conjunctival epithelial cells(P=0.03). However, zeaxanthin did nothave significant effect on H2 O2-induced cell death in limbal or conjunctival epithelial cells.CONCLUSION: Zeaxanthin is an effective reagent in reducing the detrimental effect of UV-radiation and oxidative stress on ocular surface epithelial cells.

Comparison of aerobic conjunctival bacterial flora in pregnant, reproductive-aged and postmenopausal women

AIM: To evaluate the effect of hormonal status on aerobic conjunctival flora in women. METHODS: One hundred fifty-eight women [reproductive-aged (n=55), pregnant (n=51), and postmenopausal (n=52)] who admitted to outpatient clinic of Obstetrics and Gynecology Department of Denizli State Hospital were enrolled. Age, body-mass index (BMI), obstetric history, cigarette smoking, drug usage, presence of systemic disease, and intraocular pressure (IOP) were recorded for each patient. The samples were taken from the lower fornix with two culture swabs and directly incubated in culture containing 5% sheep blood, eosin-methylene blue and chocolate agar. The other swab specimen was Gram stained. All growths and microscopic results were analyzed. RESULTS: The coagulase-negative Staphylococcus was the predominant organism isolated in the conjunctival samples in both three groups. The aerobic microorganism growth rate for all isolated aerobic organisms revealed no significant change in the three groups (P>0.05). The conjunctival culture positivity rates were similar in the three groups (49% in reproductive-aged, 57% in pregnant and 58% in postmenopausal women) (P>0.05). Age, IOP, BMI, gravidity, parity, cigarette smoking, drug usage, and presence of systemic diseases did not have an effect on culture positivity in three groups. CONCLUSION: Results of this study showed that conjunctival aerobic flora and bacterial colonization did not differ between reproductive-aged, pregnant and postmenopausal women.