Toxicity evaluation of an essential oil mixture from the Cretan herbs thyme, Greek sage and Cretan dittany.

The importance of herbal extracts on health, which was initially based on ethnopharmacological and traditional knowledge, becomes increasingly well documented by numerous experimental and intervention studies. The daily use of beverages from different aromatic plants which becomes more popular nowadays, has been a tradition in Crete, and a habit that has been linked to the longevity seen in the island. Additionally, a certain combination of aromatic plants has been used against common cold and influenza. Interestingly, when such a mixture of essential oils from Cretan herbs (Cretan Aromatic Plants essential oil, CAPeo, from thyme, Greek sage, and Cretan dittany) was formulated, significant antiviral properties were observed in vitro and a significant reduction in the duration and severity of symptoms of patients with upper respiratory tract infections was found in a clinical study. However, since many plants extracts can exert toxic effects, toxicity issues should be properly addressed. In the present work we present an acute and sub-chronic toxicity evaluation for this mixture of aromatic plants' essential oils in rats. In fact, it is the only toxicity study for Cretan dittany. We report absence of toxicity, rendering the use of the mixture of essential oils from Cretan dittany, Greek sage and thyme as safe.

Retinal Pigment Epithelial Detachment in Age-Related Macular Degeneration.

Retinal pigment epithelial detachment is defined as a separation of the retinal pigment epithelium from the inner collagenous layer of Bruch's membrane. It is a common manifestation in both dry and wet types of age-related macular degeneration. This review aims to provide a comprehensive guide to the pathophysiology, clinical and imaging characteristics, natural course and treatment of the various types of pigment epithelial detachments in order to assist in diagnosis and management of this important feature of age-related macular degeneration.

A simple open source bioinformatic methodology for initial exploration of GPCR ligands' agonistic/antagonistic properties.

Drug development is an arduous procedure, necessitating testing the interaction of a large number of potential candidates with potential interacting (macro)molecules. Therefore, any method which could provide an initial screening of potential candidate drugs might be of interest for the acceleration of the procedure, by highlighting interesting compounds, prior to in vitro and in vivo validation. In this line, we present a method which may identify potential hits, with agonistic and/or antagonistic properties on GPCR receptors, integrating the knowledge on signaling events triggered by receptor activation (GPCRs binding to Gα,ß,γ proteins, and activating Gα , exchanging GDP for GTP, leading to a decreased affinity of the Gα for the GPCR). We show that, by integrating GPCR-ligand and Gα -GDP or -GTP binding in docking simulation, which correctly predicts crystallographic data, we can discriminate agonists, partial agonists, and antagonists, through a linear function, based on the ΔG (Gibbs-free energy) of liganded-GPCR/Gα -GDP. We built our model using two Gαs (ß2-adrenergic and prostaglandin-D2 ), four Gαi (µ-opioid, dopamine-D3, adenosine-A1, rhodopsin), and one Gαo (serotonin) receptors and validated it with a series of ligands on a recently deorphanized Gαi receptor (OXER1). This approach could be a valuable tool for initial in silico validation and design of GPRC-interacting ligands.

Membrane androgen receptors (OXER1, GPRC6A AND ZIP9) in prostate and breast cancer: A comparative study of their expression.

Accumulating evidence during the last decades revealed that androgens exert membrane-initiated actions leading to the modulation of significant cellular processes, important for cancer cell growth and metastasis (including prostate and breast), that involve signaling via specific kinases. Collectively, many nonclassical, cell surface-initiated androgen actions are mediated by novel membrane androgen receptors (mARs), unrelated to nuclear androgen receptors. Recently, our group identified the G protein coupled oxo-eicosanoid receptor 1 (OXER1) (a receptor of the arachidonic acid metabolite, 5-oxoeicosatetraenoic acid, 5-oxoETE) as a novel mAR involved in the rapid effects of androgens. However, two other membrane proteins, G protein-coupled receptor family C group 6 member A (GPRC6A) and zinc transporter member 9 (ZIP9) have also been portrayed as mARs, related to the extranuclear action of androgens. In the present work, we present a comparative study of in silico pharmacology, gene expression and immunocytochemical data of the three receptors in various prostate and breast cancer cell lines. Furthermore, we analyzed the immunohistochemical expression of these receptors in human tumor and non-tumoral specimens and provide a pattern of expression and intracellular distribution.

Nuclear localization of PD-L1: artifact or reality?

BACKGROUND: The levels of expression and membrane localization of programmed cell death ligand 1 (PD-L1), an immune checkpoint type I transmembrane glycoprotein, are related to the clinical response of anti-PD-L1/PD-1 therapy. Although the biologically relevant localization of PD-L1 is on the plasma membrane of cancer cells, it has also been reported to be in the cytoplasm and sometimes in the nucleus. Furthermore, it has been claimed that chemotherapeutics can modify PD-L1 expression and/or its nuclear localization. RESULTS: Data from our group suggest that the nuclear localization of PD-L1, and other plasma membrane proteins as well, could be an artifact resulting from inadequate experimental conditions during immunocytochemical studies. Mild detergent and rigorous fixation conditions should be used in order to preserve the membrane localization and to prevent an erroneous translocation of PD-L1 and other non-interconnected membrane proteins, such as CD24, into other cellular compartments including the nucleus, of untreated and chemotherapeutically treated breast cancer cells. CONCLUSION: We propose that well-specified and rigorously followed protocols should be applied to immunocytochemical diagnostic techniques, especially to those related to individualized diagnosis and treatment.

Long-Term Results of Combination Treatment with Single-Dose Ranibizumab plus Photodynamic Therapy for Retinal Angiomatous Proliferation.

PURPOSE: To present functional and anatomic outcomes of combination therapy with ranibizumab and photodynamic therapy (PDT) in a series of retinal angiomatous proliferation (RAP) cases. METHODS: A total of 17 eyes of 17 patients with RAP were included. Thirteen eyes were treatment naïve and 4 were already on ranibizumab monotherapy. Combined treatment with single-dose ranibizumab and PDT was performed on all treatment-naïve cases. Follow-up was performed every month for 6-38 months, using optical coherence tomography and indocyanine green angiography when necessary. RESULTS: Overall, 6 out of 7 treatment-naïve cases with stage I or II disease manifested complete angiographic resolution of the hot spot with a single injection of ranibizumab plus PDT. In stage III disease, 7 out of 10 eyes showed persistent leakage immediately after the combined treatment. The rate of hot spot occlusion was found to correlate with the stage of disease (p = 0.05). There was a significant improvement in posttreatment BCVA for all patients regardless of disease stage (p = 0.02), which was more evident in the earlier stages. Improvement was also greater for treatment-naïve patients (p = 0.03). The posttreatment improvement in BCVA was strongly correlated with hot spot occlusion (p < 0.001). CONCLUSIONS: Single-dose modified anti-VEGF treatment in combination with PDT may provide long-term regression at the initial stages of RAP.

Anti-vascular endothelial growth factor for unilateral acute idiopathic maculopathy.

PURPOSE: To describe a case of unilateral acute idiopathic maculopathy (UAIM) response to intravitreal therapy with aflibercept (Eylea). METHODS: Retrospective case report. RESULTS: A 36-year-old woman with sudden visual impairment and central scotoma was found to have a UAIM in her left eye. Three weeks after continuous worsening of her visual acuity and central scotoma, the patient was treated with intravitreal injections of aflibercept. The visual acuity increased and the macula lesion regressed, causing macula scarring after 2 injections. CONCLUSIONS: Intravitreal injections of aflibercept could be a therapy option in patients with UAIM without signs of spontaneous resolution of the clinical manifestations and visual improvement.

Comparison of Dexamethasone Intravitreal Implant with Conventional Triamcinolone in Patients with Postoperative Cystoid Macular Edema.

AIMS: To evaluate the efficacy of the treatment with intravitreal triamcinolone acetonide or dexamethasone intravitreal implant in patients with postoperative cystoid macular edema (PCME). MATERIALS AND METHODS: Thirty eyes of 29 patients with PCME were randomized into two groups: one group initially received an injection of 4 mg triamcinolone; retreatment after 3 months was dependent on functional and anatomic outcome in a PRN regimen. The second group received a single injection of the dexamethasone intravitreal implant (Ozurdex). Patients were followed for 6 months. The main outcomes were best-corrected visual acuity (BCVA) and central millimeter retinal thickness (CMMT). RESULTS: Mean BCVA improved significantly in both groups at 3 months (p ≤ 0.05) and 6 months (p ≤ 0.05) after treatment. There was no statistically significant difference between the two groups in visual acuity improvement at 3 months (p > 0.05) or 6 months (p > 0.05). Mean CMMT of both groups also decreased significantly after treatment at 3 and 6 months (both p ≤ 0.05) and the reduction was significantly superior in the triamcinolone group compared to ozurdex group at 1 week and 6 months (p ≤ 0.05). All cases with intraocular hypertension were managed with IOP-lowering medication and no surgery was required during the study. One patient was excluded because of endophthalmitis in the triamcinolone group. CONCLUSION: Intravitreal triamcinolone and dexamethasone implant are both equally effective in increasing visual acuity in patients with PCME at a 6-month follow-up. However, macular edema seems to respond more rapidly with intravitreal triamcinolone, and 3-monthly repetitive injections maintain the reduction in retinal thickness better than a single dexamethasone implant at the first 6 months of follow-up period.

Ophthalmic Metastasis of Breast Cancer and Ocular Side Effects from Breast Cancer Treatment and Management: Mini Review.

Breast cancer is one of the most common malignant diseases occurring in women, and its incidence increases over the years. It is the main site of origin in ocular metastatic disease in women, and, due to its hematogenous nature of metastatic spread, it affects mainly the uveal tissue. The purpose of this paper is to summarize the clinical manifestations of the breast cancer ocular metastatic disease, alongside the side effects of the available treatment options for the management and regression of the systematic and ophthalmic disease.

The role of fundus autofluorescence imaging in the study of the course of posterior uveitis disorders.

BACKGROUND: To evaluate the correlation of fundus autofluorescence (FAF) with indocyanine green angiography (ICGA) in patients with various posterior uveitis disorders. METHODS: Interventional case series including 23 eyes of 15 patients with diagnosis of a specific type of retinochoroiditis, such as acute posterior multifocal placoid pigment epitheliopathy (APMPPE), serpiginous-like choroiditis, multifocal choroiditis (MFC), Harada disease, and syphilitic retinochoroiditis. Also, some cases with undefined retinochoroiditis were included. FAF and ICGA were performed and correlated at baseline and during follow-up after treatment. RESULTS: In ICGA, early hypofluorescence was found to be the hallmark of acute choroidal inflammation, resolving in later stages and remaining in the late phase in areas with retinal pigment epithelium (RPE) damage. Poorly defined hyperautofluorescent areas correlated with acute choroidal lesions. Hypoautofluorescent delineation suggested the initiation of RPE healing processes, correlating well with the late phase of ICGA and delineating the RPE damage. Early hyperautofluorescence with late hypofluorescence in ICGA indicated the presence of primary RPE involvement. CONCLUSION: FAF contributes to the interpretation of RPE disease and may be a useful tool for the follow-up of progressive inflammatory disorders. Comparative evaluation of FAF and ICGA allows a characterization of the sequence of inflammatory events and the level of tissue affected.

Evaluation of single-dose azithromycin versus standard azithromycin/doxycycline treatment and clinical assessment of regression course in patients with adult inclusion conjunctivitis.

BACKGROUND: Single-dose azithromycin (AZT) has been proved efficient in treating various human Chlamydia infections. However, it has not been thoroughly tested in patients with adult inclusion conjunctivitis (AIC). It is the aim of this study to perform a comparative evaluation of efficacy and safety of one-day AZT with long-term AZT and doxycycline (DOX) regimens in AIC and to present a clinical profile of regression course of the disease. MATERIALS: Eighty-three consecutive adults, with symptoms and signs of chronic conjunctivitis and positive Polymerase Chain Reaction (PCR) for chlamydia, were randomly assigned in four treatment groups; AZT 1-day 1000 mg orally, AZT 500 mg daily 9 and 14 days and DOX 200 mg 21 days orally. Follow-up visits were scheduled 1 and 2 weeks, 1, 3 and 6 months after treatment completion. PCR was repeated at the 2nd post-treatment week to confirm elimination of infectious agent. Detailed record of subjective symptoms and objective signs was performed at all visits. Retreatment rate among groups was evaluated as primary outcome. Regression rate of symptoms/signs among groups was recorded as secondary outcomes. RESULTS: All treatment groups provided statistically equivalent results of retreatment rate. Statistically significant regression of symptoms/signs was documented, initially from the 1st post-treatment week in general, but 1 month was required for complete patients' relief. Follicles were the most common clinical sign with the earliest regression after successful treatment. CONCLUSION: Single-dose azithromycin should be considered as equally reliable treatment option, comparing to long-term alternative regimens for AIC. Patients should wait for one week, until first signs of significant regression become obvious and should consider approximately one month to total relief. Follicles could be reasonably used as a key sign for clinical assessment of treatment success.

Twelve months of follow-up after intravitreal injection of ranibizumab for the treatment of idiopathic parafoveal telangiectasia.

AIMS: To report the anatomic and functional outcomes of intravitreal ranibizumab in idiopathic parafoveal telangiectasia (IPT). MATERIAL AND METHODS: Four eyes of three patients were included in this interventional case series. One patient (two eyes) had bilateral IPT (type 2) and two patients (two eyes) had unilateral (type 1) IPT. Retreatment was scheduled in case of leakage persistence in combination with visual acuity (VA) deterioration. Fluorescein angiography and optical coherence tomography were performed together with a full ophthalmic examination at baseline, 1, 3, 6, 9, and 12 months after injection. RESULTS: One intravitreal injection of ranibizumab was performed in all four eyes. Complete cessation of leakage was documented postintervention in three eyes and partial cessation in one eye, followed by improvement of best corrected VA in one of them. In all eyes, structural changes of the photoreceptor layer were detected in tomography and were responsible for visual loss, which was in most cases, refractory to the applied therapy. CONCLUSION: Use of ranibizumab might be efficient in eliminating leakage activity in the macular region in patients with IPT. Nevertheless, improvement in VA was infrequent. Preexisting early photoreceptor alteration in IPT might render such patients unable to improve VA.

Evaluation of direct immunofluorescence assay and cytological examination in comparison to polymerase chain reaction of conjunctival swabs in patients with adult inclusion conjunctivitis.

PURPOSE: To evaluate PCR, direct immunofluorescence assay (DIA) and cytological test of conjunctival swabs for the diagnosis of adult follicular conjunctivitis (AFC). METHODS: Eighty-three adult patients with chronic conjunctivitis and sixteen healthy individuals were included. Conjunctival scrapings underwent PCR, DIA and cytological analysis. Exams were repeated two weeks after treatment application. Sensitivity, specificity and agreement rate with PCR of DIA and Cytology were evaluated and correlated with clinical symptoms/signs. RESULTS: Cytology test was more sensitive than DIA and presented an acceptable agreement with PCR (K=0.44) in treatment-naïve patients, concerning especially the combination of both conventional exams (K = 0.77). Inferior diagnostic performance of was detected post-treatment, considering the combination as well (K=0.40). Negative post-treatment PCR correlated well with significant relief of symptoms/signs. CONCLUSION: Combination of Cytology and DIA seems to be a useful diagnostic option for treatment naïve AFC patients. However, PCR remains the most reliable test for post-treatment evaluation.

Human herpes virus-6 as a cause of recurrent posterior uveitis in a HIV-positive patient.

PURPOSE: To report a case of bilateral recurrent posterior uveitis caused by human herpes virus-6 (HHV-6) in a human immunodeficiency virus-positive individual. METHODS: Comprehensive ophthalmic examination, including imaging with optical coherence tomography, fluorescein and indocyanine green angiography, and adequate laboratory tests were performed. A human immunodeficiency virus-positive patient without any AIDS defining condition, with a history of recurrent bilateral posterior uveitis referred to us with the diagnosis of retinal detachment. RESULTS: Vitreous polymerase chain reaction detected an aberrant band for herpes viruses, which proved to be human herpes virus-6 by repeated polymerase chain reactions. Serum antibodies titer was positive for human herpes virus-6. The patient responded well to antiviral therapy with valacyclovir. CONCLUSION: This is the first case of human herpes virus-6-related bilateral posterior uveitis in a human immunodeficiency virus-positive individual without clinical manifestations of AIDS.

Evaluation of segmentation procedures using spectral domain optical coherence tomography in exudative age-related macular degeneration.

PURPOSE: The purpose of this study was to evaluate standardized automated segmentation procedures of spectral domain optical coherence tomography (SD-OCT) in imaging of age-related macular degeneration. METHODS: Twenty-nine eyes of 29 patients with neovascular age-related macular degeneration were included. Three groups were assigned, according to the predominant localization of extravasated fluid in the intra-, subretinal, and subretinal pigment epithelium (RPE) compartment. Automated segmentation procedures were evaluated in B scans of 512 × 128 × 1024 and 200 × 200 × 1024 scan patterns using SD-OCT (Cirrus). Alignment errors at the internal limiting membrane, actual RPE, and extrapolation of the physiologic RPE (RPE fit) were graded using a standardized classification system. RESULTS: The rate of severe alignment failures was 56% and 41% for the 512 × 128 and the 200 × 200 raster pattern, respectively. Internal limiting membrane and actual RPE boundaries were most correctly delineated in the 200 × 200 raster pattern. Retinal pigment epithelium fit alignment was generally poor in 50% of scans. Retinal thickness values defined by internal limiting membrane and actual RPE segmentation were 90% accurate and not compromised by RPE fit misalignment. Subretinal fluid was demarcated most reliably. Alignment errors may occur together with a large spectrum of morphologic alterations. CONCLUSION: Automated algorithms of SD-OCT demonstrate a substantial rate of alignment failures in the assessment of exudative age-related macular degeneration pathologies, which are usually associated with misinterpretation of boundaries at the (sub) RPE level.

Correlation of high-definition optical coherence tomography and fluorescein angiography imaging in neovascular macular degeneration.

PURPOSE: To correlate the morphologic characteristics of choroidal neovascular lesions (CNV) in age-related macular degeneration (AMD) using raster scanning high-definition optical coherence tomography (HD-OCT) and conventional fluorescein angiography (FA). METHODS: In this comparative clinical study, 37 consecutive patients with classic, minimally classic, or occult CNV; 13 patients with early AMD; and 10 age-matched healthy individuals were included. HD-OCT imaging (Topcon, Tokyo, Japan) and FA (scanning retinal ophthalmoscope; HRA2; Heidelberg Engineering, Dossenheim, Germany) were performed after a complete standardized ophthalmic examination. Only one eye of each patient was included in the study. A point-to-point correlation between HD-OCT and FA images was performed. Early and late FA images at defined locations were correlated with OCT measurements, including 3D maps, 2D single scans, a thickness linear graph, and the 3D retinal pigment epithelium (RPE) segmentation. RESULTS: With HD-OCT imaging used to delineate the lesion morphology, early AMD was detected as having a normal foveal contour and minimal alteration in the macular area; classic CNV as a well-defined lesion with steep margins and a craterlike configuration, occult CNV as an ill-defined, flat lesion with a convex surface; and minimally classic CNV as having classic and occult components. FA-OCT overlay images provided a significant correlation between FA patterns and OCT features such as retinal thickness (RT). CONCLUSIONS: 3D-OCT provided realistic anatomic maps of the retina, RPE, and RT in patients with AMD. Discrimination between the predominant CNV lesion types was achieved, and their precise shape was identified, together with information about the lesion's localization and leakage activity.

Identification of optical density ratios in subretinal fluid as a clinically relevant biomarker in exudative macular disease.

PURPOSE: To investigate the potential role of optical density ratios (ODRs) obtained from subretinal fluid analysis in exudative macular disease and to identify the predictive role of ODRs under therapy in comparison to conventional morphometric measurements (CMMs). METHODS: Fifteen patients with neovascular age-related macular degeneration (nAMD) and 15 with acute central serous chorioretinopathy (CSC) were included in this prospective comparative and interventional case series. High-definition optical coherence tomography (SD-OCT) was performed according to a standardized protocol. nAMD patients received a standard treatment consisting of three monthly doses of intravitreous ranibizumab. Best corrected visual acuity (BCVA) was assessed at baseline (BSL) and weeks 2, 4, and 12. SD-OCT parameters were compared between CSC and nAMD at baseline. Predictive factors for functional recovery under ranibizumab treatment were identified in patients with nAMD. RESULTS: ODR showed highly significant differences between CSC and nAMD, whereas it was not possible to differentiate between these diseases on the basis of CMM. During follow-up, CMM correlated with BCVA at BSL only, whereas ODR showed a significant correlation with BCVA at week 4 and 12 during antiangiogenic therapy. CONCLUSIONS: Results suggest that CMM may correlate with BCVA at BSL, but has limited predictive value regarding recovery of visual function. Most interesting, ODR correlated with BCVA under therapy and was the only parameter that was pathognomic for nAMD in contrast to CSC in this study. ODR may reflect the status of the blood-retina barrier and may be used for pathophysiologic differentiation and prognostic purposes in exudative macular disease.