Twenty-four-month real-life treatment outcomes of polypoidal choroidal vasculopathy versus type 1 macular neovascularization in Caucasians.

BACKGROUND: To compare 24-month real-world outcomes of Vascular Endothelial Growth Factor (VEGF) inhibitors for Polypoidal Choroidal Vasculopathy (PCV) and type 1 Macular Neovascularization (MNV) in a Caucasian population. METHODS: Retrospective analysis from a prospectively designed observational database. Data from Italian centres participating in the Fight Retinal Blindness! (FRB!) project were collected. Treatment-naïve PCV or type 1 MNV commencing treatment after January 2009 were included. The primary outcome was 24-month visual acuity (VA) change; other outcomes included baseline characteristics, number of anti-VEGF injections, time to lesion inactivation and proportion of active visits. RESULTS: A total of 322 eyes (114 PCVs) from 291 patients were included. Median [Q1, Q3] VA at baseline was comparable (70 [55, 75.8] vs. 70 [58.8, 75] letters, p = 0.95). Adjusted VA change at 2 years was higher in PCV (mean [95% CI], +1.2 [-1.6, 4.1] vs. -3.6 [-6, -1.2] letters, p = 0.005). PCV received fewer anti-VEGF injections over the first 24 months of treatment than type 1 MNV (median [Q1, Q3], 8 [5, 10] vs. 9 [7, 12.2] injections, p = 0.001), inactivated earlier (median [Q1, Q3], 235 [184, 308] vs. 252 [169, 343] days, p = 0.04) and was less frequently graded 'active' (62% vs. 68% of visits, p = 0.001). CONCLUSIONS: PCV had slightly better VA outcomes over 24 months of treatment than type 1 MNV after receiving less anti-VEGF injections. These results suggest a possible overlap of the two clinical entities with similar visual prognosis in Caucasians.

Neuroprotective potential of Laurus nobilis antioxidant polyphenol-enriched leaf extracts.

Oxidative stress has been proposed to be an important factor in the pathogenesis of Alzheimer's disease (AD), playing a central role in amyloid ß-protein (Aß) generation and neuronal apoptosis. Oxidative damage directly correlates with the presence of Aß deposits. Aß and oxidative stress jointly induce neuronal death, Aß deposits, gliosis, and memory impairment in AD. In order to counteract AD neurodegeneration, the inhibition of the vicious cycle of Aß generation and oxidation is an attractive therapeutic strategy, and antiamyloidogenic and antioxidant herbal drugs could represent an alternative and valid approach. In this context, an alcoholic extract from Laurus nobilis leaves (LnM) and seven fractions obtained therefrom were of interest. All extracts prepared through extractive and chromatographic techniques were phytochemically studied by chromatographic techniques including gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS(n)). The potential antioxidant efficacy of the obtained fractions was screened by DPPH(•) and ABTS(•+) assays, as well as specific assay media characterized from the presence of highly reactive ROS and RNS species (ROO(•), OH(•), O2(•-), and NO). In order to evaluate the preparation of safe and nontoxic extracts, MTT, SRB, and LDH assays toward SH-5YSY and SK-N-BE(2)-C human neuronal cell lines, as well as on C6 mouse glial cell line, were performed. The apoptosis-inducing properties by spectroscopic evaluation of the extracts' ability to activate caspase-3 and by a DNA fragmentation assay were also investigated. Data thus obtained allowed us to state the absence of toxic effects induced by phenolic-rich fractions (LnM, LnM-1, LnM-1a, LnM-1b, and LnM-2c), which at the same time exerted significant cytoprotective and antioxidant responses in hydrogen peroxide and Aß(25-35)-fragment-oxidized cell systems. The potential antiamyloidogenic efficacy of Laurus nobilis leaf polar extracts in the Aß(25-35) fragment oxidized cell systems was further analyzed by Congo red staining.

Subretinal fluid may protect against macular atrophy in neovascular age-related macular degeneration: 5 years of follow-up from Fight Retinal Blindness registry.

PURPOSE: The purpose of the study was to assess the association of macular atrophy (MA) according to the activity of macular neovascularization (MNV) (inactive, only subretinal fluid [SRFL], or active, i.e. including intraretinal fluid [IRFL]) using optical coherence tomography (OCT) in patients with neovascular age-related macular degeneration (nAMD). METHODS: Multicentric observational study. Treatment-naïve nAMD eyes without subfoveal MA or subretinal fibrosis (SF) at baseline were included since 1st January 2010 and 30th September 2016 to allow up to 5 years of treatment follow-up. Eyes were grouped based on their predominant activity status as: (1) mostly inactive, (2) mostly active non-SRFL only [IRFL] or (3) mostly active-SRFL only [onlySRFL]. Kaplan-Meier survival curves estimated the time to development of MA or SF. Cox proportional hazards models evaluated predictors of developing subfoveal MA or SF. The main outcome measure was the risk of developing MA according to predominant MNV activity. RESULTS: A total of 973 eyes were eligible for analysis. OnlySRFL eyes had lower risk of developing subfoveal MA (HR [95% CI]: 0.56 [0.36, 0.88]; p = 0.024) and extrafoveal MA (HR [95% CI]: 0.41 [0.27, 0.61]; p < 0.001) than IRFL eyes. IRFL eyes had lower visual acuity (VA) (54.5 letters) and the highest proportion of eyes with vision ≤35 letters (25%) at 5 years while onlySRFL eyes had comparable 5-year VA (63.7 letters) to inactive eyes (63.7 letters). CONCLUSION: Subretinal fluid appears to protect against MA. Distinguishing the compartment of retinal fluid and understanding its relationship with MA and SF can guide the management of nAMD.

A tandem mass spectrometric investigation of the low-energy collision-activated fragmentation of neo-clerodane diterpenes.

Mass spectrometric fragmentation data of neo-clerodane diterpenes are almost inexistent but they can prove helpful for the qualitative and quantitative analysis of these compounds as well as for the identification of unknown compounds belonging to this class of plant secondary metabolites. [M-H](-) ions of nine neo-clerodane diterpenes (1-9), recently isolated from Teucrium chamaedrys, were generated by electrospray ionization and were fragmented in the collision cell of a Triple Quadrupole (TQ) and of a Quadrupole Ion Trap (QIT) mass spectrometer. The deprotonated neo-clerodane glucosides, chamaedryoside A and B (1, 2), readily lost the sugar residue to give, as their main fragmentation channel, the neo-clerodane ions, I and II, which were structurally characterized by TQ and QIT MS. The collision-activated dissociation (CAD) mass spectra of I and II and of deprotonated neo-clerodanes 3-9 allowed us to reach some general conclusions on the fragmentation pathways of this class of compounds. For example, teuflin and its OH derivatives, teucrin A, teuflidin and 6-beta-hydroxyteucridin, showed a characteristic fragmentation pattern involving the loss of 94 Da and 124 Da from the lactone moiety, whereas a loss of 44 Da was observed for teucrin E, and of 58 Da for teucrin F and G. In addition, several compound-specific fragmentations were observed and can be proposed for the identification of individual compounds. The systematic approach allowed us to hypothesize the mechanisms of the most important collision-activated dissociation/isomerization channels.

Structural discrimination of isomeric tetrahydrofuran lignan glucosides by tandem mass spectrometry.

Due to the possible role in human health, the number of analytical studies on lignans aimed at their quali- and quantitative analysis in plant extracts, biological fluids and foods is continuously increasing. However, helpful systematic mass spectrometric investigations on these compounds are few and rather limited to specific lignan sub-classes. To increase the comprehension of the previously outlined picture of the gas-phase properties of furofuran lignans, we extended the study to tetrahydrofuran lignans and here we reported the collision-activated dissociation (CAD) fragmentation patterns of the alkali metal cation adducts, [M+Alk](+), and [M-H](-) ions of three isomeric tetrahydrofuran lignans, (+)-8'-hydroxylariciresinol 4'-O-beta-D-glucopyranoside (1), (+)-7'-hydroxylariciresinol 7'-O-beta-D-glucopyranoside (2) and 4-O-beta-D-glucopyranosyloxy-3,3'-dimethoxy-7,9'-epoxylignan-5',8',9-triol (3) investigated by electrospray ionization triple quadrupole mass spectrometry (ESI-TQMS). Hydrogen/deuterium (H/D) solution exchange experiments, allowing the selective H/D exchange of all the acidic hydrogen atoms, proved to be a very effective tool to obtain information on the nature of fragments generated during TQ/CAD processes. The [M+Na](+) CAD mass spectra of the three isomeric tetrahydrofurans revealed four different pathways involving the loss of the glucose moiety, which allowed the assignment of the glycosylation site. In the negative ion mode, the main fragmentation channel of the [M-H](-) ions of O-glucosylated lignans at the phenolic oxygen atoms is represented by the loss of 162 Da. When the sugar is bound to a benzylic OH group the loss of the sugar as a 180 Da unit occurs eventually following the loss of a water molecule involving both the C(9)H(2)OH chain and the sugar.

abeo-abietanes from Teucrium polium roots as protective factors against oxidative stress.

Six new 17(15→16)-abeo-abietanes, along with seven known compounds, were isolated and characterized from ethyl acetate root extract of Teucrium polium L., a medicinal plant belonging to the Labiatae family reported to have hypolipidemic, hypoglycaemic, anti-nociceptive and anti-inflammatory effects. Their structures were elucidated by 1D ((1)H, (13)C and DEPT) and 2D (COSY, TOCSY, HSQC, HMBC, and NOESY) NMR and mass spectral data. The antioxidative properties of pure metabolites were analyzed on the basis of their DPPH radical scavenging capability. The antioxidant capacity in cell-free systems of the isolated metabolites was carried out by measuring their capabilities to inhibit the synthesis of thiobarbituric acid reactive species in assay media using as oxidable substrates a vegetable fat and the pentose sugar 2-deoxyribose and to prevent oxidative damage of the bovine serum albumin (BSA) hydrosoluble protein. All of the compounds showed a significant and dose-response efficacy although weaker than that exercised by the standard Trolox®.

Structure determination of chamaedryosides A-C, three novel nor-neo-clerodane glucosides from Teucrium chamaedrys, by NMR spectroscopy.

Three new nor-neo-clerodane diterpenes, named chamaedryoside A (1), B (2) and C (3), have been isolated from the organic extracts of Teucrium chamaedrys (L.) and their structural characterization has been accomplished by (1)H and (13)C-NMR spectra, and DEPT, by COSY, TOCSY, HSQC, HSQC-TOSCY and HMBC experiments, as well as by ESI-MS/MS techniques. The stereostructures have been elucidated by NOESY and computational calculations.

Antioxidant properties of sour cherries (Prunus cerasus L.): role of colorless phytochemicals from the methanolic extract of ripe fruits.

Many edible plant metabolites are known to be useful as cellular antioxidants. In the search for antioxidative chemicals from native fruits of the Campania region of Italy, Prunus cerasus L., an acidic cherry widely used for culinary purposes, has been studied. Fruit crude extracts (MeOH, EtOAc, and hexane) were submitted to an antioxidative screening using specific assay media characterized from the presence of highly reactive radical species (DPPH*, ABTS*+, O2*-, NO) or lipoperoxidation markers. The reducing power of the samples was also determined. It was observed that the most polar extracts in MeOH and EtOAc were able to exercise a massive and dose-increasing antioxidative capacity. The peculiar efficacy of the same extracts was revealed by investigating their protein and deoxyribose oxidation capacity. A preliminary analysis of total phenol, flavonoid, and anthocyanin contents together with biological screening data put the basis on P. cerasus fruit phytochemical investigation of methanolic extract. Twenty secondary metabolites were isolated and characterized by spectroscopic (especially 1D and 2D NMR) and spectrometric techniques. 1-(4-Hydroxyphenyl)-1,2-ethanediol-1,2-bis-1-O-beta-D-glucopyranoside (3), (4-hydroxy-3-methoxyphenyl)methanol-1-O-beta-D-gentiobioside (4), epicatechin-3-malate (14), and epicatechin-3-(1''-methyl)malate (15) were isolated for the first time. All of the compounds were evaluated for their radical scavenging activity on DPPH*, O2*-, and NO. Flavonoids and quinic acid derivatives were found to be the more antioxidative substances.

Isolation and structure elucidation of antioxidant polyphenols from quince (Cydonia vulgaris) peels.

Thirteen new compounds, as well as 16 already known, have been isolated from organic extracts of peels of Cydonia vulgaris, a fruit of a shrub belonging to the same tribe as the apple. All of the structures were elucidated by EI- or ESI-MS and (1)H and (13)C NMR after purification of individual compounds by HPLC. Thirteen fatty acid esters of cinnamyl alcohols, three fatty acid esters of hydroxybenzoic acid, three fatty acid esters of hydroxybenzaldehyde, three glucosides of aromatic acids, four chlorogenic acids, two flavonols, and a benzylamine have been identified. The fatty acid moieties have been identified by GC-MS analysis of the methanolysis products. All of the compounds were tested for their radical scavenging and antioxidant activities by measuring their capacity to scavenge the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical and anion superoxide radical and to induce the reduction of Mo(VI) to Mo(V). The chlorogenic acids and the flavonols exhibited more antioxidant and radical scavenger capacity than the positive standards alpha-tocopherol and ascorbic acid. The results of the tests were analyzed by cluster analysis that grouped all of the compounds on the basis of the substituents on the aromatic ring.

Chemical characterization of new oxylipins from Cestrum parqui, and their effects on seed germination and early seedling growth.

Isolation, chemical characterization, and phytotoxicity of five new oxylipins, together with seven already known related compounds, from Cestrum parqui L' Hérl. is reported. All the structures were elucidated on the basis of their spectral data, especially 1D-(1H- and 13C-NMR, DEPT) and 2D-NMR (COSY, TOCSY, HSQC, HMBC, and NOESY). The configurations of the stereogenic C-atoms were determined by the Mosher's method. The compounds have been assayed for their phytotoxicity on Lactuca sativa at concentrations ranging between 10(-4) and 10(-8) M. The results of the phytotoxicity tests on the germination and growth of the test species, obtained by a cluster analysis, showed interesting relationship between the chemical structures of the compounds and their biological effects.

Isolation, structure elucidation, and antioxidant evaluation of cydonioside A, an unusual terpenoid from the fruits of Cydonia vulgaris.

A novel terpenoid, cydonioside A (1), was isolated from the fruits of Cydonia vulgaris Pers. Its structure and relative configuration were elucidated on the basis of in-depth spectroscopic analyses, including 2D-NMR experiments as well as MM+ calculations. Compound 1 was assayed for its radical-scavenging activity towards the DPPH radical and the superoxide radical anion (O2*-), as well as for its overall antioxidant activity, as assessed by the formation of a phosphomolybdenum complex.

Reactive oxygen species scavenging activity of flavone glycosides from Melilotus neapolitana.

One new and six known flavone glycosides were isolated from the MeOH extract of Melilotus neapolitana Ten. The new compound, identified as 7-O-beta-D-glucopyranosyloxy-4',5-dihydroxy-3-[O-alpha-L-rhamnopyranosyl-(1-->6)-3-O-beta-D-glucopyranosyloxy]flavone (1) by 1D and 2D NMR techniques and mass spectra, was isolated along with kaempferol-3-O-rutinoside (2), kaempferol-3-O-glucoside (3), rutin (4), quercetin-3-O-glucoside (5), isorhamnetin-3-O-rutinoside (6), and isorhamnetin-3-O-glucoside (7). The antioxidant and radical scavenging activities of these compounds and the whole crude methanol extract were evaluated. The organic extract can inhibit MDA marker's synthesis by 57%. All the metabolites displayed good reducing power, with the kaempferol (2,3) and isorhamnetin derivatives (6,7) being less active than the corresponding quercetin derivatives 4,5.

Triamcinolone as adjunctive treatment to laser panretinal photocoagulation for proliferative diabetic retinopathy.

OBJECTIVE: To evaluate intravitreal injection of triamcinolone acetonide before laser panretinal photocoagulation (PRP) in the treatment of proliferative diabetic retinopathy. METHODS: This interventional case series included 9 patients with bilateral proliferative diabetic retinopathy. One eye received intravitreal triamcinolone before PRP (injected eye) and the other, PRP alone (control eye). The main outcome measures were the change in planimetric area of fluorescein leakage from retinal neovascularization and in central macular thickness on optical coherence tomography at 3, 6, 9, and 12 months. Secondary end points were change in vision, intraocular pressure, and cataract progression. RESULTS: All patients completed 9 months and 5 patients, 12 months of follow-up. Initial mean (SD) planimetric area of fluorescein leakage and central macular thickness were 7.22 (5.70) mm(2) and 372.11 (91.88) microm in injected eyes and 9.08 (6.17) mm(2) and 355.33 (115.23) microm in control eyes, respectively. At the 9- and 12-month intervals, the planimetric area of fluorescein leakage decreased by 86% and 88% in injected eyes and 33% and 50% in controls, respectively. Central macular thickness significantly decreased in injected eyes and increased in control eyes. Vision slightly improved in injected eyes and worsened in control eyes. CONCLUSION: Intravitreal injection of triamcinolone before PRP may be useful in improving the effects of PRP in eyes with proliferative diabetic retinopathy by reducing neovascularization and macular thickening.

Structures of bioactive carexanes from the roots of Carex distachya Desf.

Four metabolites, named carexanes I-L, have been isolated from the roots of Carex distachya Desf, an herbaceous plant living in the Mediterranean maquis, together with three known compounds, already isolated from the aerial part of the plant. All the compounds have been characterized on the basis of their spectroscopic properties. Carexane I derived from the lose of a proton from the C-18 carbon of an intermediate isopropyl cation. Its stereostructure has been elucidated by Mosher's method, NOESY/ROESY experiments and computational calculations. The bioactivity on seed germination and root/shoot growth of Lactuca sativa L. of all the isolated compounds is also reported.

Spectroscopic identification and antioxidant activity of glucosylated carotenoid metabolites from Cydonia vulgaris fruits.

Carotenoid metabolites are common plant constituents with significant importance for the flavor and aroma of fruits. Three new carotenoid derivatives, (2E,4E)-8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 1-O-beta-D-glucopyranosyl ester (1), (2Z,4E)-8-beta-D-glucopyranosyloxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid (3), and 3,9-dihydroxymegastigmast-5-ene-3-O-[beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (5), as well as three known compounds, have been isolated from the ethanolic extract of peels of Cydonia vulgaris, the fruit of a shrub belonging to the same family as the apple. All the compounds were identified by spectroscopic techniques, especially 1D and 2D NMR. Antioxidant activities of all the isolated metabolites were assessed by measuring their ability to scavenge DPPH radical and superoxide radical (O2*-) and to induce the reduction of Mo(VI).

Influence of harvest season on chemical composition and bioactivity of wild rue plant hydroalcoholic extracts.

The rue (Ruta graveolens) copiousness in rural areas of the Campania Region based a thorough chemical and biological investigation aimed at exploring the seasonal variability of phenol constituents in rue leaves and its influence on their antioxidant, cytotoxic and anti-inflammatory capabilities. To this purpose, hydroalcoholic extracts were prepared from plant samples seasonally collected. LC-ESI-MS/MS techniques were employed to analyze qualitatively and quantitatively the seasonal rue phenol content, whereas different chemical antioxidant assays (by DPPH, ABTS, Fe(3+) RP, ORAC, and FCR methods) and XTT redox metabolic activity assay were performed to screen the seasonal phenol complex-related antioxidant and cytotoxic power. The ability of the rue leaf extracts to counteract cyclooxygenase-2 (COX-2) expression was also evaluated. Data obtained highlighted that the adopted extraction procedure markedly pauperized the furanocoumarin content in all the prepared rue extracts. Flavonol glycosides, along with the flavone acacetin and two sinapic acid derivatives were the main constituents of the spring harvest-derived extract, which exerted the highest antioxidant capability in cell-free systems and was capable to inhibit COX-2 synthesis by 44% comparably to dexamethasone, used as positive control. Data provide new insights for developing a proper management of rue plants for new safe industrial purposes in herbal medicine field.

Structural characterization of phytotoxic terpenoids from Cestrum parqui.

Isolation, chemical characterization and phytotoxicity of nine polyhydroxylated terpenes (five C13nor-isoprenoids, two sesquiterpenes, a spirostane and a pseudosapogenin) from Cestrum parqui L'Herr are reported. In this work we completed the phytochemical investigation of the terpenic fraction of the plant and described the structural elucidation of polar isoprenoids using NMR methods. All the configurations of the compounds have been assigned by NOESY experiments. Four new structures have been identified as (3S,5R,6R,7E,9R)-5,6,9-trihydroxy-3-isopropyloxy-7-megastigmene, 5alpha-spirostan-3beta,12beta,15alpha-triol, and 26-O-(3'-isopentanoyl)-beta-d-glucopyranosyl-5alpha-furost-20(22)-ene-3beta,26-diol, and as an unusual tricyclic sesquiterpene. The compounds have been assayed for their phytotoxicity on lettuce at the concentrations ranging between 10(-4) and 10(-7)M. The activities of some compounds were similar to that of the herbicide pendimethalin.

Isolation, characterization, and antioxidant activity of E- and Z-p-coumaryl fatty acid esters from cv. Annurca apple fruits.

A total of 12 fatty acid esters of Z- and E-p-coumaryl alcohol were isolated from cv. Annurca apple fruit and characterized. This apple variety is widely cultivated in the south of Italy, and the fruits typically undergoe a reddening treatment after harvest. Structures of the p-coumaryl esters were elucidated by GC-MS and (1)H and (13)C NMR after purification of individual compounds by HPLC. It was found that the esters are localized in the fruit peel. During reddening of the fruit, there was a substantial increase in the amount of esters and particularly in molecular species with unsaturated fatty acids. The individual compounds were tested for antioxidant activity, and over half were shown to be at least as effective as alpha-tocopherol.

Radical-scavenging activities of new hydroxylated ursane triterpenes from cv. Annurca apples.

Two new ursolic acid triterpene derivatives, compounds 2 and 3, have been isolated from cv. Annurca apple fruit, a high-quality apple variety widely cultivated in southern Italy, together with the known 2-oxopomolic acid (1). The new compounds were identified by means of different spectroscopic techniques as 3-epi-2-oxopomolic acid (= (3alpha)-3,19-dihydroxy-2-oxours-12-en-28-oic acid; 2) and (1alpha)-1-hydroxy-3-oxours-12-en-28-oic acid (3). Compounds 1-3 were tested for their radical-scavenging activities with the aid of a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (Fig. 2). All three constituents showed activities similar to that of the reference antioxidant alpha-tocopherol (vitamin E).

1H NMR based metabolic profiling of eleven Algerian aromatic plants and evaluation of their antioxidant and cytotoxic properties.

Eleven Algerian medicinal and aromatic plants (Aloysia triphylla, Apium graveolens, Coriandrum sativum, Laurus nobilis, Lavandula officinalis, Marrubium vulgare, Mentha spicata, Inula viscosa, Petroselinum crispum, Salvia officinalis, and Thymus vulgaris) were selected and their hydroalcoholic extracts were screened for their antiradical and antioxidant properties in cell-free systems. In order to identify the main metabolites constituting the extracts, 1H NMR-based metabolic profiling was applied. Data obtained emphasized the antiradical properties of T. vulgaris, M. spicata and L. nobilis extracts (RACI 1.37, 0.97 and 0.93, respectively), whereas parsley was the less active as antioxidant (RACI -1.26). When the cytotoxic effects of low and antioxidant doses of each extract were evaluated towards SK-N-BE(2)C neuronal and HepG2 hepatic cell lines, it was observed that all the extracts weakly affected the metabolic redox activity of the tested cell lines. Overall, data strongly plead in favor of the use of these plants as potential food additives in replacement of synthetic compounds.