A Protocol for microbiologically safe preparation, storage, and use of autologous serum eye-drops in low-income countries

Introduction: The study aimed to investigate whether the preparation, storage, and use of autologous serum in insulin syringes is microbiologically safe. Methodology: Blood samples (10 mL) were obtained from 10 volunteers. After centrifugation, the supernatant serum was removed and distributed in 5 sterile insulin syringes for each sample; syringes were numbered 0 to 4 and labelled with the subject’s details. Syringes were immediately transported to the microbiology laboratory and stored in a refrigerator at +4°C. The “0” labelled syringes were separated from the others and 100 μl of serum from each syringe was immediately seeded on chocolate and Sabouraud agar plates, which were incubated aerobically at 37°C for 96 hours to detect any bacterial and/or fungal contamination. In the next 4 days, the same procedure was repeated for the remaining syringes: on day 1, the “1” labelled syringes were analyzed; on day 2, the “2” labelled ones, and so on. In a second experiment, blood samples were obtained from 5 different volunteers. The same procedure as above was followed, but each syringe was used for repeated cultures at 2-hour intervals, for a total of 12 cultures/day. The needle was removed and replaced for each inoculation and the syringes were stored in the refrigerator after use. Results: Under these experimental conditions, none of the cultures showed microbial growth. Conclusions: Results suggest that, under the protocol described, preparation, storage and use of undiluted autologous serum in insulin syringes is inexpensive, fast, and microbiologically safe. This is of great importance for low-income countries.</br

Detection of serum antibodies cross-reacting with <i>Mycobacterium avium</i> subspecies <i>paratuberculosis</i> and beta-cell antigen zinc transporter 8 homologous peptides in patients with high-risk proliferative diabetic retinopathy

Purpose: MAP3865c, a Mycobacterium avium subspecies paratuberculosis (MAP) cell membrane protein, has a relevant sequence homology with zinc transporter 8 (ZnT8), a beta-cell membrane protein involved in Zn++ transportation. Recently, antibodies recognizing MAP3865c epitopes have been shown to cross-react with ZnT8 in type 1 diabetes patients. The purpose of this study was to detect antibodies against MAP3865c peptides in patients with high-risk proliferative diabetic retinopathy and speculate on whether they may somehow be involved in the pathogenesis of this severe retinal disorder. Methods: Blood samples were obtained from 62 type 1 and 80 type 2 diabetes patients with high-risk proliferative diabetic retinopathy and 81 healthy controls. Antibodies against 6 highly immunogenic MAP3865c peptides were detected by indirect ELISA. Results: Type 1 diabetes patients had significantly higher rates of positive antibodies than controls. Conversely, no statistically significant differences were found between type 2 diabetes patients and controls. After categorization of type 1 diabetes patients into two groups, one with positive, the other with negative antibodies, we found that they had similar mean visual acuity (&#8764;0.6) and identical rates of vitreous hemorrhage (28.6%). Conversely, Hashimoto's thyroiditis prevalence was 4/13 (30.7%) in the positive antibody group and 1/49 (2%) in the negative antibody group, a statistically significant difference (P = 0.016). Conclusions: This study confirmed that type 1 diabetes patients have significantly higher rates of positive antibodies against MAP/ZnT8 peptides, but failed to find a correlation between the presence of these antibodies and the severity degree of high-risk proliferative diabetic retinopathy. The significantly higher prevalence of Hashimoto's disease among type 1 diabetes patients with positive antibodies might suggest a possible common environmental trigger for these conditions

Combined Phacoemulsification and Intravitreal Dexamethasone Implant (Ozurdex®) in Diabetic Patients with Coexisting Cataract and Diabetic Macular Edema

Purpose. To investigate the effectiveness and safety of combined phacoemulsification and dexamethasone intravitreal implant in patients with cataract and diabetic macular edema. Methods. In this two-center, retrospective, single-group study, the charts of 16 consecutive patients who underwent combined phacoemulsification and intravitreal dexamethasone implant were retrospectively reviewed. These 16 patients, 7 men and 9 women, were observed at least 3 months of follow-up. Primary outcome was the change of the central retinal thickness (CRT); secondary outcome was the change of best-corrected visual acuity (BCVA). Any ocular complications were recorded. Results. Mean CRT decreased significantly from 486 ± 152.4 μm at baseline to 365.5 ± 91 μm at 30 days (p=.005), to 326 ± 80 μm at 60 days (p=.0004), and to 362 ± 134 μm at 90 days (p=.001). Mean BCVA was 20/105 (logMAR, 0.72 ± 0.34) at baseline and improved significantly (p≤.007) at all postsurgery time points. One case of ocular hypertension was observed and successfully managed with topical therapy. No endophthalmitis or other ocular complications were observed. Conclusion. Intravitreal slow-release dexamethasone implant combined with cataract surgery may be an effective approach on morphologic and functional outcomes for patients with cataract and diabetic macular edema for at least three months after surgery

Bromfenac eyedrops in the treatment of diabetic macular edema: a pilot study

PURPOSE: To evaluate the efficacy and safety of topical bromfenac in patients with newly diagnosed diabetic macular edema (DME). METHODS: In this pilot study including 17 patients with monocular, newly diagnosed DME, diagnosis of DME was established by the detection of retinal thickening at or within 500 μm of the center of the macula on ophthalmoscopic examination, according to the Early Treatment Diabetic Retinopathy Study classification. Central macular thickness (CMT) was determined by optical coherence tomography. Bromfenac sodium hydrate 0.9 mg/mL eyedrops were administered in the affected eye twice daily for 30 days. Primary endpoints were changes in best-corrected visual acuity (BCVA) and CMT at the end of therapy. RESULTS: Topical bromfenac significantly reduced mean CMT, from 465.41 ± 118.47 μm at baseline to 388.88 ± 152.63 μm posttreatment (p = 0.02). There was no significant change in BCVA and differences in mean macular volume fell just short of statistical significance (p = 0.06). Treatment was well-tolerated, and there were no topical or systemic side effects. CONCLUSIONS: Topical bromfenac twice daily may play a role in the reduction of DME. These preliminary results warrant further larger multicenter studies to confirm our findings and establish whether topical bromfenac may be of long-term benefit in the treatment of DME

Bromfenac eyedrops in the treatment of diabetic macular edema: a pilot study

PurposeTo evaluate the efficacy and safety of topical bromfenac in patients with newly diagnosed diabetic macular edema (DME).MethodsIn this pilot study including 17 patients with monocular, newly ..

Plasmapheresis, Intravenous Immunoglobulins, and Autologous Serum Eyedrops in the Acute Eye Complications of Toxic Epidermal Necrolysis

PURPOSE: Toxic epidermal necrolysis (TEN) is a rare, life-threatening, drug-induced, mucocutaneous disease, which can severely affect the ocular surface. The purpose of this study was to investigate the efficacy of plasmapheresis, human IV immunoglobulins (IVIg), and autologous serum (AS) eyedrops in the treatment of the severe acute ocular complications of TEN. METHODS: A retrospective chart review of all patients admitted to the Burn Unit, Azienda Ospedaliero-Universitaria-Sassari, Sassari, Italy, from 2009 to 2015, identified 9 patients (2 men, 7 women; mean age 63.8 ± 24.7 years) with TEN. Bilateral, acute ocular surface complications were observed in 7 (78%) patients; 3 showed catarrhal conjunctivitis, whereas 4 had severe pseudomembranous conjunctivitis and corneal ulcers. RESULTS: All patients with TEN were immediately treated with plasmapheresis and human IVIg, which produced a marked improvement in the patients' general condition. In the 3 with catarrhal conjunctivitis, preservative-free artificial tears and topical antibiotics were beneficial. In the 4 with severe pseudomembranous conjunctivitis and corneal ulcers, treatment with AS eyedrops resulted in corneal and conjunctival epithelium healing over 3-6 weeks. After a minimum follow-up of at least 12 months, there were minimal/mild residual signs and symptoms of dry eye. CONCLUSIONS: Plasmapheresis and IVIg may be life-saving and contribute to reduce ocular surface inflammation in TEN. Autologous serum eyedrops, prepared after plasmapheresis completion and IVIg infusion, may be helpful in the management of the severe acute ocular complications of TEN

Plasmapheresis, intravenous immunoglobulins, and autologous serum eyedrops in the acute eye complications of toxic epidermal necrolysis

PURPOSE: Toxic epidermal necrolysis (TEN) is a rare, life-threatening, drug-induced, mucocutaneous disease, which can severely affect the ocular surface. The purpose of this study was to investigate the efficacy of plasmapheresis, human IV immunoglobulins (IVIg), and autologous serum (AS) eyedrops in the treatment of the severe acute ocular complications of TEN. METHODS: A retrospective chart review of all patients admitted to the Burn Unit, Azienda Ospedaliero-Universitaria-Sassari, Sassari, Italy, from 2009 to 2015, identified 9 patients (2 men, 7 women; mean age 63.8 ± 24.7 years) with TEN. Bilateral, acute ocular surface complications were observed in 7 (78%) patients; 3 showed catarrhal conjunctivitis, whereas 4 had severe pseudomembranous conjunctivitis and corneal ulcers. RESULTS: All patients with TEN were immediately treated with plasmapheresis and human IVIg, which produced a marked improvement in the patients' general condition. In the 3 with catarrhal conjunctivitis, preservative-free artificial tears and topical antibiotics were beneficial. In the 4 with severe pseudomembranous conjunctivitis and corneal ulcers, treatment with AS eyedrops resulted in corneal and conjunctival epithelium healing over 3-6 weeks. After a minimum follow-up of at least 12 months, there were minimal/mild residual signs and symptoms of dry eye. CONCLUSIONS: Plasmapheresis and IVIg may be life-saving and contribute to reduce ocular surface inflammation in TEN. Autologous serum eyedrops, prepared after plasmapheresis completion and IVIg infusion, may be helpful in the management of the severe acute ocular complications of TEN

Iritis and angle closure glaucoma caused by caterpillar hairs

Purpose To report on a child presenting with iritis and angle closure glaucoma caused by caterpillar hairs. Methods A 12-year-old girl complained of sudden pain and visual loss in her left eye while she was playing in a pinewood. Two hours later, she developed maculo-papular rash on her limbs. On examination, left visual acuity was 1/10. The affected eye showed conjunctival hyperemia, corneal edema, anterior chamber flare (4+ cells). Corneal staining with fluorescein disclosed diffuse superficial punctate keratopathy. Left IOP was 55 mm Hg. Visante OCT of the affected eye showed a thin, translucent, linear foreign body into the cornea and anterior chamber up to the irido-corneal angle. Topical steroids, antibiotics, cycloplegics, hypotensive eyedrops, and intravenous mannitol were given. Results Inspection of the place where the girl was playing revealed the presence of numerous processionary pine caterpillars. On the basis of this finding and the clinical data, a diagnosis of iritis and angle closure glaucoma caused by caterpillar hairs was made. After 3 days’ treatment, corneal edema and anterior chamber inflammation resolved, IOP returned to normal, and visual acuity was 10/10 again. However, a small iris cyst was still visible on OCT scans. Conclusion Ocular lesions caused by caterpillar hairs are uncommon. Ophthalmologists should be aware that intense iritis with angle closure glaucoma may be the result of the penetration of caterpillar hairs into the eye. OCT scans may be important to confirm the diagnosis.</br

Plasmapheresis and autologous serum eye-drops in the treatment of acute ocular complications from toxic epidermal necrolysis (Lyell syndrome)

Purpose Toxic epidermal necrolysis (TEN) is a rare, life-threatening, drug-induced, mucocutaneous disease, which can affect the ocular surface. The purpose of this study was to evaluate the acute ocular complications from TEN and investigate on the efficacy of plasmapheresis and serum-eyedrops in the treatment of this condition. Methods A retrospective chart review of all Burn Unit patients admitted from 2009-2013 identified 7 TEN patients (2 men, 5 women; mean age: 69.6±20.3 years). TEN was associated with allopurinol use in 4 patients, with gefitinib in 1, and with trimethoprim/sulphamethoxazole in 1. In the remaining patient, the causative agent was not identified. Acute bilateral ocular complications were observed in 5 patients; 3 showed mild ocular surface inflammation, whereas 2 had severe pseudomembranous conjunctivitis with corneal ulcers. All TEN patients were immediately treated with plasmapheresis. Results In the 2-3 weeks after plasmapheresis, there was a marked improvement of the patients' general condition. In those with mild ocular surface inflammation, the use of preservative-free artificial tears and steroids was beneficial. In the 2 with pseudomembranous conjunctivitis and corneal ulcers, treatment with autologous serum eye-drops and preservative-free artificial tears was effective, resulting in corneal epithelium healing and mild conjunctival scarring. Conclusion Not all TEN patients have ocular complications at onset. Plasmapheresis may be life-saving and contribute to reduce ocular surface inflammation. Autologous serum eye-drops prepared after plasmaferesis may be helpful in the management of the acute ocular complications caused by TEN.</br