Personalized Pseudophakic Model for Refractive Assessment

Purpose

To test a pseudophakic eye model that allows for intraocular lens power (IOL) calculation, both in normal eyes and in extreme conditions, such as post-LASIK.

Methods

Participants: The model’s efficacy was tested in 54 participants (104 eyes) who underwent LASIK and were assessed before and after surgery, thus allowing to test the same method in the same eye after only changing corneal topography.

Modelling

The Liou-Brennan eye model was used as a starting point, and biometric values were replaced by individual measurements. Detailed corneal surface data were obtained from topography (Orbscan®) and a grid of elevation values was used to define corneal surfaces in an optical ray-tracing software (Zemax®). To determine IOL power, optimization criteria based on values of the modulation transfer function (MTF) weighted according to contrast sensitivity function (CSF), were applied.

Results

Pre-operative refractive assessment calculated by our eye model correlated very strongly with SRK/T (r = 0.959, p<0.001) with no difference of average values (16.9±2.9 vs 17.1±2.9 D, p>0.05). Comparison of post-operative refractive assessment obtained using our eye model with the average of currently used formulas showed a strong correlation (r = 0.778, p<0.001), with no difference of average values (21.5±1.7 vs 21.8±1.6 D, p>0.05).

Conclusions

Results suggest that personalized pseudophakic eye models and ray-tracing allow for the use of the same methodology, regardless of previous LASIK, independent of population averages and commonly used regression correction factors, which represents a clinical advantage.     

Two Odorant-Binding Proteins Mediate the Behavioural Response of Aphids to the Alarm Pheromone (E)-?-farnesene and Structural Analogues

Background

Aphids are agricultural pests of great economical interest. Alternatives to insecticides, using semiochemicals, are of difficult applications. In fact, sex pheromones are of little use as aphids reproduce partenogenetically most of the time. Besides, the alarm pheromone, (E)-ß-farnesene for a great number of species, is difficult to synthesize and unstable in the environment. The search for novel semiochemicals to be used in population control can be efficiently approached through the study of the olfactory system at the biochemical level. Recently odorant-binding proteins (OBPs) have been shown to play a central role in olfactory recognition, thus becoming the target of choice for designing new semiochemicals.

Methodology/Principal Findings

To address the question of how the alarm message is recognised at the level of OBPs, we have tested 29 compounds, including (E)-ß-farnesene, in binding assays with 6 recombinant proteins and in behaviour experiments. We have found that good repellents bind OBP3 and/or OBP7, while non repellents present different spectra of binding. These results have been verified with two species of aphids, Acyrthosiphon pisum and Myzus persicae, both using (E)-ß-farnesene as the alarm pheromone.

Conclusions

Our results represent further support to the idea (so far convincingly demonstrated only in Drosophila) that OBPs are involved in decoding the chemical information of odorants and pheromones, and for the first time provide such evidence in other insect species and using wild-type insects. Moreover, the data offer guidelines and protocols for the discovery of potential alarm pheromones, using ligand-binding assays as a preliminary screening before subjecting selected compounds to behaviour tests.     

Epidemiology of dermatophytoses: retrospective analysis from 2005 to 2010 and comparison with previous data from 1975

Dermatophyte infections are extremely frequent worldwide and their epidemiological features vary according to the geographical area and have changed in the last decades. We studied the spectrum of dermatophytoses by means of a retrospective analysis involving 6,133 patients referred to the Mycology Service of the Dermatology Clinic of Policlinico Hospital - University of Bari, Italy during the period 2005-2010. The most frequent clinical forms were tinea unguium (39.2% of the total dermatophytoses), tinea corporis (22.7%) and tinea pedis (20.4%). There was a predominance of women for tinea unguium and corporis and of men for tinea pedis and especially tinea cruris. T. rubrum was the prevalent causative agent, implicated in 64% of total cases, followed by M. canis (14%) and T. mentagrophytes (10%). The retrospective evaluation of epidemiological data collected at our Clinic since 1975 showed a gradual decrease in the frequency of tinea cruris, tinea corporis, and tinea capitis over time. On the contrary, during the past two decades, there has been a progressive increase in the frequency of tinea pedis and especially of tinea unguium. In parallel with this changing pattern, the frequency of isolation of T. rubrum has shown a continuous increase during the last 35 years, whereas a progressive decline of the etiological role of T. violaceum, M. canis and even more of E. floccosum has been noted.     

Emergence and characterisation of vanB vancomycin-resistant Enterococcus faecalis in Rio de Janeiro, Brazil

Here we describe the detection and characterisation of three isolates of vancomycin-resistant VanB-type Enterococcus faecalis. Sequence analysis suggested that these isolates harboured the vanB1 gene. The isolates were susceptible to the majority of antimicrobial agents tested, with the exception of chloramphenicol, erythromycin and vancomycin, and showed distinct profiles of high-level resistance to aminoglycosides. Analysis of the clonal relatedness of the vanB E. faecalis isolates showed similar pulsed-field gel electrophoresis profiles. To our knowledge, this is the first report of the occurrence of enterococcal strains carrying vanB genes in Brazil.     

Purification and Biological Activities of Abelmoschus esculentus Seed Lectin

The Abelmoschus esculentus (Malvaceae) plant originated in Africa and has spread across a number of tropic countries, including northeastern Brazil. The plant has been used to treat various disorders, such as cancer, microbial infections, hypoglycemia, constipation, urine retention and inflammation. The lectin of A. esculentus (AEL) was isolated by precipitation with ammonium sulfate at a saturation level of 30/60 and purified by ion exchange chromatography (Sephacel-DEAE). The electrophoresis (SDS-PAGE) profile of the AEL showed two protein bands of apparent molecular mass of approximately 15.0 and 21.0?kDa. The homogenity of the protein was confirmed by electrospray mass spectrometry (ESI-MS), which revealed the presence of a 10.29-kDa monomer and a 20.58-kDa dimer. The AEL exhibits agglutinating activity against rabbit (74.41 UH/mP) and human type ABO erythrocytes (21.00 UH/mP). This activity does not require the presence of divalent cations and is specifically inhibited by lactose, fructose and mannose. The intravenous treatment with 0.01, 0.1 and 1?mg/kg of AEL inhibited the paw edema elicited by carrageenan by approximately 15, 22 and 44?%, respectively, but not that induced by dextran. In addition, treatment with 0.1, 1 and 10?mg/kg of AEL also inhibited the abdominal writhing induced by acetic acid by approximately 52, 57 and 69?%, respectively. In conclusion, AEL is a new lectin with a molecular mass of 20.0?kDa, which is -composed of a 10.291-Da monomer and a 20.582-kDa dimer, that exhibits anti-inflammatory, antinociceptive and hemagglutinating activities. In addition, the lectin hemagglutinating property is both metallo-independent and associated with the lectin domain.     

An investigation of the ligand binding properties and negative cooperativity of soluble insulin-like growth factor receptors

To investigate the interaction of the insulin-like growth factor (IGF) ligands with the insulin-like growth factor type 1 receptor (IGF-1R), we have generated two soluble variants of the IGF-1R. We have recombinantly expressed the ectodomain of IGF-1R or fused this domain to the constant domain from the Fc fragment of mouse immunoglobulin. The ligand binding properties of these soluble IGF-1Rs for IGF-I and IGF-II were investigated using conventional ligand competition assays and BIAcore biosensor technology. In ligand competition assays, the soluble IGF-1Rs both bound IGF-I with similar affinities and a 5-fold lower affinity than that seen for the wild type receptor. In addition, both soluble receptors bound IGF-II with similar affinities to the wild type receptor. BIAcore analyses showed that both soluble IGF-1Rs exhibited similar ligand-specific association and dissociation rates for IGF-I and for IGF-II. The soluble IGF-1R proteins both exhibited negative cooperativity for IGF-I, IGF-II, and the 24-60 antibody, which binds to the IGF-1R cysteine-rich domain. We conclude that the addition of the self-associating Fc domain to the IGF-1R ectodomain does not affect ligand binding affinity, which is in contrast to the soluble ectodomain of the IR. This study highlights some significant differences in ligand binding modes between the IGF-1R and the insulin receptor, which may ultimately contribute to the different biological activities conferred by the two receptors.     

Detection of mutagens in water-distribution systems after disinfection

This research examined the quality of water-before and after distribution-of four drinking-water production plants located in Northern Italy, two of which collected water from local aquifers and two from the River Po. A battery of genotoxicity assays for monitoring drinking-water was performed to assess the quality of the water produced by the treatment plants under study. Three different sampling stations were selected at each plant, one right at the outlet of the treatment plant and two along with the distribution pipelines. Raw river water was also sampled and analysed as a control. The water samples (500 l) were concentrated on silica C18 cartridges and the extracts were tested in in vitro mutagenicity assays (Salmonella/microsome assay with strains TA 98 and TA 100; SOS Chromotest with Escherichia coli strain PQ37); gene conversion, point mutation and mitochondrial DNA mutability assays with the diploid Saccharomyces cerevisiae strain D7 and a toxicity test using the bioluminescent bacterium Vibrio fischeri (Microtox). The Microtox test and the mitochondrial DNA mutability assay showed the greatest sensitivity towards toxic or mutagenic substances in the water extracts considered. The results show that this battery of short-term tests is applicable in the routine monitoring of drinking-water quality before and after distribution.     

Purification of polyhydroxybutyrate produced by <Emphasis Type="Italic">Burkholderia cepacia</Emphasis> IPT64 through a chemical and enzymatic route

This work aimed at the study of purification of biopolymers produced by Burkholderia cepacia IPT64, through a chemical attack like an adjuvant procedure through chemical and enzymatic route. Enzymatic lysis using protease was run on an enzyme/cell ratio of 0.02. Chemical attacks were performed as pre or post-treatment for enzymatic attacks, using sodium dodecyl sulphate or hydrogen peroxide. The same chemicals and procedures were used alone as a control. Fenton’s reagent was also tested as a chemical treatment. Using only one of the chemicals H₂O₂, Fenton’s reagent or SDS, the purity increase achieved values of 14%, 16% or 23%, respectively. When H₂O₂ or SDS were used as pre-treatment for the enzymatic attack, the results of purity increase achieved values of 58% for H₂O₂/cell ratio between 0.60 and 1.20, and 57% when SDS/cell ratio at 0.56 was used. In the case when H₂O₂ or SDS were used as post-treatment for the enzymatic attack, results of purity increase achieved 60% when the H₂O₂ was used at H₂O₂/cell ratio ranging between 0.30 and 0.60 and 71% when SDS was used at a ratio SDS/cell of 0.56.     

Mixed-linkage (1-->3),(1-->4)-beta-D-glucan is not unique to the Poales and is an abundant component of Equisetum arvense cell walls

Mixed-linkage (1-->3),(1-->4)-beta-D-glucan (MLG) is widely considered to be a defining feature of the cell walls of plants in the Poales order. However, we conducted an extensive survey of cell-wall composition in diverse land plants and discovered that MLG is also abundant in the walls of the horsetail Equisetum arvense. MALDI-TOF MS and monosaccharide linkage analysis revealed that MLG in E. arvense is an unbranched homopolymer that consists of short blocks of contiguous 1,4-beta-linked glucose residues joined by 1,3-beta linkages. However, in contrast to Poaceae species, MLG in E. arvense consists mostly of cellotetraose rather than cellotetriose, and lacks long 1,4-beta-linked glucan blocks. Monosaccharide linkage analyses and immunochemical profiling indicated that, in E. arvense, MLG is a component of cell walls that have a novel architecture that differs significantly from that of the generally recognized type I and II cell walls. Unlike in type II walls, MLG in E. arvense does not appear to be co-extensive with glucuroarabinoxylans but occurs in walls that are rich in pectin. Immunofluorescence and immunogold localization showed that MLG occurs in both young and old regions of E. arvense stems, and is present in most cell types apart from cells in the vascular tissues. These findings have important implications for our understanding of cell-wall evolution, and also demonstrate that plant cell walls can be constructed in a way not previously envisaged.