Soluble Cyanobacterial Carotenoprotein as a Robust Antioxidant Nanocarrier and Delivery Module

To counteract oxidative stress, antioxidants including carotenoids are highly promising, yet their exploitation is drastically limited by the poor bioavailability and fast photodestruction, whereas current delivery systems are far from being efficient. Here we demonstrate that the recently discovered nanometer-sized water-soluble carotenoprotein from Anabaena sp. PCC 7120 (termed AnaCTDH) transiently interacts with liposomes to efficiently extract carotenoids via carotenoid-mediated homodimerization, yielding violet–purple protein samples. We characterize the spectroscopic properties of the obtained pigment–protein complexes and the thermodynamics of liposome–protein carotenoid transfer and demonstrate the delivery of carotenoid echinenone from AnaCTDH into liposomes with an efficiency of up to 70 ± 3%. Most importantly, we show efficient carotenoid delivery to membranes of mammalian cells, which provides protection from reactive oxygen species (ROS). Incubation of neuroblastoma cell line Tet21N in the presence of 1 μM AnaCTDH binding echinenone decreased antimycin A ROS production by 25% (p < 0.05). The described carotenoprotein may be considered as part of modular systems for the targeted antioxidant delivery.BMBF, 01DJ15007, Carotenoidbindende photoschaltbare Proteine: Lichtinduzierte Dynamik und Anwendungen in modernen mikroskopischen Verfahre

Rational management of acute keratoconus

<p>Acute keratoconus is a common and severe complication of advanced progressive keratoconus that occurs in more than 30 % of cases. Acute corneal edema in advanced progressive keratectasia is reffered to as acute corneal hydrops (hydrops corneae). It has been also reported in other ectatic disorders such as pellucid marginal degeneration. The most common misdiagnosis in hydrops is HSV disciform keratitis or acute bacterial keratitis. 126 corneal hydrops patients (79 men, 47 women) aged 16‑63 (129 eyes) were observed and treated over the last five years. 124 patients were diagnosed with acute keratoconus and 2 patients were diagnosed with pellucid marginal degeneration. Acute kereatoconus patients<br />represented a special and compromised cohort with systemic allergic diseases (neurodermatitis and various atopic disorders, n = 48), Down’s syndrome (n = 16) or mental disorders (n = 19). In many of these patients who vigorously rubbed their eyes, keratectasia progressed more rapidly. In 7 cases, acute keratoconus developed during pregnancy. 3 cases of recurrent keratoconus were reported — in a woman with Down’s syndrome (recurrence in 3 years), in a man with severe neurodermatitis (recurrence in 5 years), and in a man with anamnestic acute keratoconus (recurrence in 20 years). 3 patients experienced bilateral acute keratoconus. Acute keratoconus can be subdivided by the area of corneal edema into three categories, i.e., partial (6 mm or less, 52 eyes), subtotal (7‑10 mm, 56 eyes), and total (more than 10 mm, 21 eyes). Corneal edema ultimately disappeared, however, acute keratoconus resulted in a deep local scarring through the corneal layers. Slit lamp exam revealed Descemet’s membrane ruptures (so-called fish mouth). 73 eyes were referred to refractive penetrating keratoplasty (PKP). Corneal perforation was unusual even in severe corneal thinning (4 cases). In one case, descemetocele with a high risk of perforation was observed. 4 eyes required subtotal PKP. In one case, linear perforation was closed with interrupted sutures. Postoperative period following PKP was often complicated. Medical therapy which included topical steroids (eye drops and injections) in combination with antibiotics and drugs that promote corneal healing (Corneregel, Hylo-PARIN, VitA-POS) for 2‑3 months provided more rapid symptom relief and visual rehabilitation. There are many conflicting approaches to manage keratoconus. However, our long-term experience argues for PKP after complete symptoms relief excepting corneal perforation.</p

Pellucid Marginal Corneal Degeneration and Keratoconus: Differential Diagnosis and Management of Patients

The authors present the problem of differential diagnosis of primary corneal keratectasias: pellucid marginal degeneration (PMD) and keratoconus. Some cases of differential diagnosis can be very challenging and choice of rational treatment for these patients is actual. Despite multiple publications concerning keratoconus and extensive knowledge of this disease, articles about PMD are rare. Results of surgical treatment for PMD are usually worse than for keratoconus. The authors show the value of confocal microscopy, keratotopography, and high-resolution optical tomography of the anterior segment of the eye based on a two-channel Scheimpflug camera and give clinical examples of differential diagnosis that present certain complexity. Recommendations on treatment of patients with PMD are given, including such severe complication of progressive far-advanced corneal ectatic diseases as acute corneal hydrops. The important role of modern scleral gas-permeable contact lenses in visual correction of PMD and keratoconus is summarized. The authors believe that in cases of PMD without critical thinning of the cornea, if patients are satisfied with contact correction, the surgical treatment should be avoided or postponed

Enucleaton of the right eye due to large choroidal melanoma with simultaneous penetrating cornea transplantation from OD to OS (Case report).

There is presentation case report of 75‑year old woman with choroidal melanoma (T3N0M0) on the right eye and failed graft on the left pseudophakic eye with far advanced glaucoma and ARMD. No treatment was given to the leading eye with VA 0,2. VA of OS = 1 / ∞ pr.l.certa; PKP OS in 2008 for pseudophakic bullous keratopathy on the eye with far advanced glaucoma. IOP was normal after previous filtering surgery. After PKP VA = 0,04; clear graft during 2 years; then gradually opacification and vascularization occurred. VA dropped to light perception. Echography OD — tumor h 8,29 mm, d 21,77 mm. No ingrowth of tumor into anterior segment of the eye; VA OD = 0 (no light perception). Concerning the need to enucleate the right eye with large choroidal melanoma, the advantage of using corneal autograft from OD to OS, location of tumor in the posterior pole with no ingrowth in anterior segment, the decision was made to perform the following operation — to enucleate the right eye and transplant simultaneously corneal graft from OD on OS. Patient was discharged from the Ophthalmology Hospital with VA OS = 0,01, during next week VA improved to 0,02. 8,0 mm graft isclear, fixed with 8 interruptured and continious suture 10 / 0‑nylon. Anterior chamber — normal depth, atrophic iris, stable position of PC IOL. Optic nerve head is pale with subtotal deep glaucomatous excavation. Conclusion: presented case report demonstrates the rarepossibility to use cornea after enucleation the eye with large malignant tumor (located in the posterior pole) for grafting in the only eye with failed vascularised graft. It was the only possibility for this patient to restore some vision

Features of Diagnostics and Complex Approach to Therapy of Herpetic Keratitis

Objective. To determine the characteristics of drug therapy in patients with herpetic keratitis (HK), based on laboratory diagnosis.Material and methods. 49 patients with clinical symptoms of HK were included in the study. The conjunctival cells were used as a material for laboratory diagnosis of adenovirus and enterovirus infection by polymerase chain reaction (PCR) and for microbiological examination. Herpesvirus DNA in tears, saliva, urine and blood was determined using PCR (for varicella-zoster virus (VZV) and human herpesvirus type 7 (HHV-7)) and real-time PCR (for herpes simplex virus types 1 and 2 (HSV-1,2), Epstein-Barr virus (VEB), cytomegalovirus (CMV) and human herpesvirus type 6 (HHV-6). Tear and urine were examined for the presence of DNA of obligate intracellular parasites (chlamydia, ureaplasma, and mycoplasma) by PCR.Results. Сlinical picture of HK was presented by superficial (57.1 %, n = 28) and deep forms of corneal inflammation (42.9 %, n = 21). The results of laboratory diagnostics of herpetic infection had showed replication of herpesviruses in all examined patients. Genetic material of herpesviruses in lacrimal fluid was found in half of the examined patients (51 %), in saliva — in 67,3 % of cases. Viruria was observed in 21 patients (42.9 %), viraemia in 6 (12.2 %). DNA of intracellular parasites in lacrimal fluid was detected in 18.4 % of patients and in urine — in 12.2 % of patients. Bacteriological sowing of the conjunctiva scrape was positive in 75.5 % of cases.Conclusion. Instrumental diagnostics (visometry, biomicroscopy) of HK should be supplemented with laboratory research of activity of not only herpesvirus infection, but also bacterial pathogens. Positive results of microbiological sowing of conjunctival scrapers allow to approach reasonably the prescription of antimicrobial drugs in patients with HK

New possibilities of excessive postoperative scarring prophylaxis by glaucoma surgery

Purpose: to access the possibilities of new biodegradable drainage implant Glautex in the original method of glaucoma surgical treatment.Methods: 152 patients (158 eyes) have had a penetrating and nonpenetrating antiglaucoma surgical treatment with the use of Glautex. Patients were divided in 3 groups: 1st group — 90 patients (93 eyes), where valve trabeculectomy with the use of biodegrad- able drainage Glautex has been performed. 2nd group — 33 patients (35 eyes) with refractory glaucoma (previously operated glau- coma, postkeratoplasty, neovascular glaucoma etc.) with the same type of antiglaucoma surgery. 3rd group — 29 patients (30 eyes) with deep non-penetrating sclerectomy with the use of Glautex.Results: Stable IOP has been obtained in all cases within 6 to 12 months’ time. Complete biodegradation of the implant took place in 4-5 months, which was confirmed by ultrasound biomicroscopy. We have obtained no cases of inflammation in postoperative period. No cases of blebitis and cystoid blebs have been noted. the choroidal detachment, requiring surgical treatment was noted on 7 eyes (7.5%) in the 1st group; in the 2nd group — on 3 eyes (8.6%). the 3rd group (nonpenetrating glaucoma surgery) had 2 cases of flat choroidal detachment, which have successfully cured after medical nonsurgical treatment.Conclusion: Use of the new biodegradable drainage implant Glautex provides stable decrease of IOP due to prevention of sclero- conjunctival and sclero-scleral adhesions. Glautex may be used in all cases of glaucoma surgery (penetrating and nonpenetrating) with the scleral flap formation. the proposed new method is applicable for all types of glaucoma

New Properties of the Heparin-Containing Drug in vitro (Potential Antiviral and Anti-Inflammatory Effects)

Purpose: to study the cytological, immunological and antiviral effects of the HILOPARIN-KOMOD® drug in vitro.Material and methods. We used transplantable cultures of normal cells of the human Chang conjunctiva, and the kidney cells of the Vero monkeys. The cytotoxic effect of the HILOPARIN-KOMOD® was  determined by the effect on the cell viability, and by optical density  (OP) of the monolayer of the Chang conjunctiva cell culture using the enzyme immunoassay (ELISA) with MTT. The effect of the drug on  the functional activity of conjunctival cells was evaluated by the  production of cytokines at the level of their in vitro transcription. The antiviral effect of the drug HILOPARIN-KOMOD® was studied on  the Vero cell line infected with herpes simplex virus type 1 (HSV-1) and HSV type 2 (HSV-2). The viral activity of HSV-1 and HSV-2  and the antiherpetic effect after the drug was evaluated by  polymerase chain reaction (PCR). Results. No significant cytotoxic effect on the metabolism of conjunctival cells of the preparation HILOPARINKOMOD ® in  dilutions from 1/2 to 1/2048 was revealed (in comparison with the  control). The effect of the drug in dilutions of 1/40 and 1/1000 on  the culture of the conjunctival cells resulted in suppression of the  production of interferon λ-1 mRNA (IFNλ-1) and IFNλ-2 mRNA  compared to the control. In the dilution of preparation 1/1000, the  production of mRNA of interleukin-6 (IL-6) was not revealed with  simultaneous presence of IL-10 mRNA. In different dilutions of the  drug, the number of copies of HSV-1 virus DNA decreased in  comparison with the control in all cases, the greatest antiviral effect  was achieved in a 1: 2 dilution. The effect of the preparation  HILOPARIN-KOMOD® on the culture of cells infected with HSV-2 in  dilutions of 1: 2 and 1: 5 led to a decrease in the level of viral replication by 300 and 40 times, respectively.The conclusion. The drug HILOPARIN-KOMOD® has an anti-inflammatory effect that manifests itself in suppressing the synthesis of the mRNA of the acute phase proinflammatory cytokine IL-6 and  stimulating the production of anti-inflammatory IL-10 mRNA, without having a cytotoxic effect on conjunctival cells. The proven antiviral  effect of the drug makes it possible to recommend the inclusion of  HILOPARIN-KOMOD® in the scheme of therapy of patients with viral ophthalmopathology

Optical Correction of Keratoconus with a Scleral Gas-Permeable Lenses

The optical properties of the cornea are determined by its ability to refract and transmit light. Keratoconus changes cornea’s shape, the surface of the cornea becomes irregular, which leads to a violation of light refraction and the occurrence of optical aberrations. The progressing course of the disease and its late detection lead to a delayed start of therapeutic measures, which affects the prognosis of the disease progression and reduces the patient’s quality of life. The quality of visual functions depends on the stage of the process. There are 4 stages of keratoconus (according to M. Asler), each of which corresponds to certain changes in refraction and degree of deformation of the cornea. Early biomicroscopic signs are: “dilution” of the stroma (inhomogeneity of the cornea and a grayish tint in the zone of the developing apex), change in the shape of endothelial cells and clearly visible nerve endings due to longitudinal thickening. In the second stage of keratoconus, the biomicroscopic picture is complemented by the appearance of keratoconus lines (Vogt’s striae). The opacities of the Bowman’s membrane indicate the beginning of the scarring process and the transition of the disease to its third stage. The fourth stage of the disease is characterized by further development of stromal opacities and the occurrence of gross changes of the Descemet’s membrane. Advanced medical equipment for topographic mapping and measuring the cornea makes it much easier for ophthalmologists to diagnose keratoconus and choose more effective treatment methods: crosslinking or surgical treatment. Later it allows to stabilize keratoconus, but does not provide high visual acuity due to the induction of optical aberrations, including high order optical aberrations. Contact lens vision correction is the main way to correct the refractive error resulting from keratoconus. However, the use of corneal gas permeable or soft contact lenses cannot provide high quality vision, additionally causing discomfort associated with their excessive mobility. The use of scleral gas permeable contact lenses is the most effective method of optical correction of all stages of keratoconus and after keratoplasty

Structural basis for the ligand promiscuity of the neofunctionalized, carotenoid-binding fasciclin domain protein AstaP

Abstract Fasciclins (FAS1) are ancient adhesion protein domains with no common small ligand binding reported. A unique microalgal FAS1-containing astaxanthin (AXT)-binding protein (AstaP) binds a broad repertoire of carotenoids by a largely unknown mechanism. Here, we explain the ligand promiscuity of AstaP-orange1 (AstaPo1) by determining its NMR structure in complex with AXT and validating this structure by SAXS, calorimetry, optical spectroscopy and mutagenesis. α1-α2 helices of the AstaPo1 FAS1 domain embrace the carotenoid polyene like a jaw, forming a hydrophobic tunnel, too short to cap the AXT β-ionone rings and dictate specificity. AXT-contacting AstaPo1 residues exhibit different conservation in AstaPs with the tentative carotenoid-binding function and in FAS1 proteins generally, which supports the idea of AstaP neofunctionalization within green algae. Intriguingly, a cyanobacterial homolog with a similar domain structure cannot bind carotenoids under identical conditions. These structure-activity relationships provide the first step towards the sequence-based prediction of the carotenoid-binding FAS1 members