X-linked cone-rod dystrophy (locus COD1): identification of mutations in RPGR exon ORF15

X-linked cone-rod dystrophy (COD1) is a retinal disease that primarily affects the cone photoreceptors; the disease was originally mapped to a limited region of Xp11.4. We evaluated the three families from our original study with new markers and clinically reassessed all key recombinants; we determined that the critical intervals in families 2 and 3 overlapped the RP3 locus and that a status change (from affected to probably unaffected) of a key recombinant individual in family 1 also reassigned the disease locus to include RP3 as well. Mutation analysis of the entire RPGR coding region identified two different 2-nucleotide (nt) deletions in ORF15, in family 2 (delAG) and in families 1 and 3 (delGG), both of which result in a frameshift leading to altered amino acid structure and early termination. In addition, an independent individual with X-linked cone-rod dystrophy demonstrated a 1-nt insertion (insA) in ORF15. The presence of three distinct mutations associated with the same disease phenotype provides strong evidence that mutations in RPGR exon ORF15 are responsible for COD1. Genetic heterogeneity was observed in three other families, including the identification of an in-frame 12-nt deletion polymorphism in ORF15 that did not segregate with the disease in one of these families.     

Antimicrobial activity of some Satureja essential oils

The genus Satureja is represented by fifteen species of which five are endemic and Satureja pilosa and S. icarica have recently been found as new records for Turkey. Aerial parts of the Satureja pilosa, S. icarica, S. boissieri and S. coerulea collected from different localities in Turkey were subjected to hydrodistillation to yield essential oils which were subsequently analysed by GC and GC/MS. The main constituents of the oils were identified, and both antibacterial and antifungal bioassays were applied. Carvacrol (59.2%, 44.8%, 42.1%) was the main component in the oils of S. icarica, S. boissieri and S. pilosa, respectively. The oil of S. coerulea contained beta-caryophyllene (10.6%) and caryophyllene oxide (8.0%) as main constituents.     

Rapid screening of solvents and carrier compounds for lactic acid recovery by emulsion liquid extraction and toxicity on Lactobacillus casei (ATCC 11443)

This paper describes a rapid method to identify the best solvent and carrier compound combinations with the highest extraction capability and the lowest microbial toxicity characteristics for product recovery from microbial fermentation. The extraction system has an aqueous phase, and an emulsion phase, which was a blend of sodium carbonate and organic phase [91% (v/v) organic solvent, 5% (v/v or wt/v) carrier compound, and 4% (v/v) surfactant Span 80]. Alamine 336, or tri-n-octylamine in n-heptane; Alamine 336, Alamine 304, or tributyl phosphate in hexane; and Alamine 304 or tributyl phosphate in iso-octane; Alamine 304 or Amberlite in xylene demonstrated high lactic acid extraction. For determination of bacterial toxicity of selected solvent and carrier compounds, Lactobacillus casei subsp. rhamnosus (ATCC 11443) was grown in LAF medium containing one of the selected organic solvent, carrier compound, and Span 80 in 250 ml flask at 37 °C and 125 rpm. Samples were collected regularly during 48 hour incubation, and measured for changes in cell density by absorbance at 620 nm, cell count using a fluorescent dye with flow cytometry, and lactic acid, and glucose concentrations by HPLC. Hexadecane:tributyl phosphate, n-dodecane:tri-n-octylamine, and kerosene:tri-n-octylphosphine oxide demonstrated the least microbial toxicity among the tested blends with excess solvent media. Whereas, hexanes:Alamine 304 and xylenes:Alamine 304 were nontoxic in solvent saturated media.This revised version was published online in October 2005 with corrections to the Cover Date.     

Body fat distribution has no effect on serum visfatin levels in healthy female subjects

Obesity is the presence of either abnormal absolute amount or relative proportion of body fat. Contrary to gluteal obesity, visceral obesity is associated with different metabolic alterations including insulin resistance (IR). A relatively new adipocytokine visfatin is shown to be expressed predominantly in visceral fat and exhibit insulin-mimicking effects in rodents. It is still unclear whether serum visfatin levels are associated with increased total or visceral fat mass in humans. The aim of our study was to investigate the relation between visfatin and obesity parameters namely body mass index (BMI) and waist circumference (WaC) and IR in healthy female subjects. Eighty one female subjects ?20 years of age, having no diagnosis of glucose intolerance or diabetes, hypertension and dyslipidemia were chosen. The patients were divided into four groups according to their BMI and WaC values. Serum visfatin and HOMA-IR levels did not differ among groups. No correlation was detected between serum visfatin levels and obesity and metabolic parameters. In conclusion, we demonstrated that body fat distribution did not affect serum visfatin levels in healthy female subjects. Further studies are needed to clarify the exact factors influencing and determining serum visfatin levels and its clinical reflections.     

Effects of coenzyme Q10 on the heart ultrastructure and nitric oxide synthase during hyperthyroidism

Coenzyme Q10 is an important component of mitochondrial electron transport chain and antioxidant. Hyperthyroidism manifests hyperdynamic circulation with increased cardiac output, increased heart rate and decreased peripheral resistance. The heart is also under the oxidative stress in the hyperthyroidism. The aim of this study was to examine both how the coenzyme Q10 can affect heart ultrastructure in the hyperthyroidism and how the relationship between nitric oxide synthase (NOS) and heart damage and coenzyme Q10. Swiss Black C57 mice received 5 mg/kg L-thyroxine. Coenzyme Q10 (1.5 mg/kg) and L-thyroxine together was given to second group mice. Coenzyme Q10 and serum physiologic were applied to another two groups, respectively. All treatments were performed daily for 15 days by gavage. Free triiodothyronine and thyroxine were increased in two groups given L-thyroxine; thyroid-stimulating hormone level did not change. Hyperthyroid heart showed an increased endothelial NOS (eNOS) and inducible NOS (iNOS) immunoreactivity in the tissue. Coenzyme Q10 administration decreased these NOS immunoreactivities in the hyperthyroid animals. Cardiomyocytes of the hyperthyroid animals was characterized by abnormal shape and invaginated nuclei, and degenerative giant mitochondria. Desmosome plaques reduced in density. In hyperthyroid mice given coenzyme Q10, the structural disorganization and mitochondrial damage regressed. However, hearts of healthy mice given coenzyme Q10 displayed normal ultrastructure, except for increased mitochondria and some of them were partially damaged. Coenzyme Q10 increased the glycogen in the cardiomyocytes. In conclusion, coenzyme Q10 administration can prevent the ultrastructural disorganization and decrease the iNOS and eNOS increment in the hyperthyroid heart.     

A selective molecularly imprinted polymer for immobilization of acetylcholinesterase (AChE): an active enzyme targeted and efficient method

In the present study, we immobilized acetylcholinesterase (AChE) enzyme onto acetylcholine removed imprinted polymer and acetylcholine containing polymer. First, the polymers were produced with acetylcholine, substrate of AChE, by dispersion polymerization. Then, the enzyme was immobilized onto the polymers by using two different methods: In the first method (method A), acetylcholine was removed from the polymer, and then AChE was immobilized onto this polymer (acetylcholine removed imprinted polymer). In the second method (method B), AChE was immobilized onto acetylcholine containing polymer by affinity. In method A, enzyme‐specific species (binding sites) occurred by removing acetylcholine from the polymer. The immobilized AChE reached 240% relative specific activity comparison with free AChE because the active enzyme molecules bounded onto the polymer. Transmission electron microscopy results were taken before and after immobilization of AChE for the assessment of morphological structure of polymer. Also, the experiments, which include optimum temperature (25–65°C), optimum pH (3–10), thermal stability (4–70°C), kinetic parameters, operational stability and reusability, were performed to determine the characteristic of the immobilized AChE. Copyright © 2015 John Wiley & Sons, Ltd.     

Altitudinal variation in the morphometric characteristics of Aedes vexans Meigen from northeastern Turkey

Body size is one of the most significant features of organisms and is correlated with a large number of ecological and physiological variables. Similar to size, biological shape is one of the most conspicuous aspects of an organism's phenotype and provides a link between the genotype and the environment. Body size may change with altitude and also habitat differences associated with altitude may affect the biological shape and some morphological characteristics. Four populations of Aedes vexans Meigen occurring in different ecological subregions at altitudes between 808–1,620 m in the Aras Valley from northeastern Turkey were compared using traditional and geometric morphometrics. When the wing shape differences of populations were analyzed by UPGM, the cluster analyses recognized two main groups of populations. Gödekli (808 m) comprised the first group while Zülfikar (848 m), Sürmeli (944 m), and Cilehane (1,620 m) populations comprised the second group. In the second group, Zülfikar and Cilehane populations showed a similar grouping pattern while Sürmeli appeared as a different group. Centroid sizes were used as measures of overall wing size differences among different regions. Aedes vexans from the Sürmeli region had relatively larger wings.     

Miniaturized lensless imaging systems for cell and microorganism visualization in point-of-care testing

Low-cost, robust, and user-friendly diagnostic capabilities at the point-of-care (POC) are critical for treating infectious diseases and preventing their spread in developing countries. Recent advances in micro- and nanoscale technologies have enabled the merger of optical and fluidic technologies (optofluidics) paving the way for cost-effective lensless imaging and diagnosis for POC testing in resource-limited settings. Applications of the emerging lensless imaging technologies include detecting and counting cells of interest, which allows rapid and affordable diagnostic decisions. This review presents the advances in lensless imaging and diagnostic systems, and their potential clinical applications in developing countries. The emerging technologies are reviewed from a POC perspective considering cost effectiveness, portability, sensitivity, throughput and ease of use for resource-limited settings.     

Evaluating the addition of activated carbon to heat-treated mushroom casing for grain-based and compost-based substrates

Two substrates, a non-composted grain spawn substrate and a traditional composted substrate, each covered with peat-based casing that contained varying amounts of activated carbon (AC) and each receiving different heat-treatment durations, were tested for Agaricus bisporus mushroom production. The amounts of AC were 0, 5, 10, 15, and 20% v/v, and the heat treatments were 0, 60, and 180 min at 121 °C and 103.4 kPa. Overall, the addition of AC up to 10–15% of casing for a grain spawn substrate increased mushroom yield. However, the addition of AC to the casing for compost substrates had no significant effect on yield, whereas heat-treating the casing increased yield. The onset of fruiting was retarded in grain spawn treatments not receiving AC with heat-treatment durations of 60 and 180 min, whereas this effect was not as apparent for the compost substrates. On average, mushroom yield was greater for the grain spawn substrate (366 g) than for compost substrate (287 g). For grain spawn substrate, the results show that the addition of AC ranging from 5% to 10% was adequate for maximum mushroom production.     

Evaluation of culture medium for nisin production in a repeated-batch biofilm reactor

In this study, a biofilm reactor with plastic composite support (PCS), made by high-temperature extrusion of agricultural products and polypropylene, was evaluated for nisin production using L. lactis strain NIZO 22186. The high-biomass density of the biofilm reactor was found to contribute to a significantly shorter lag time of nisin production relative to a suspended-cell reactor. In comparison to glucose (579 IU/mL), sucrose significantly increased the nisin production rate by 1.4-fold (1100 IU/mL). However, results revealed that high levels of sucrose (8% w/v) had a suppressing effect on nisin production and a stimulating effect on lactic acid production. A high concentration of MgSO4.7H2O at 0.04% (w/v) was found to reduce the nisin production, while concentrations of KH2PO4 of up to 3% (w/v) did not have any significant effect on growth or nisin production. The best of the tested complex media for nisin production using the PCS biofilm reactor consisted of 4% (w/v) sucrose, 0.02% (w/v) MgSO4.7H2O, and 0.1% (w/v) KH2PO4. Nisin production rate in the biofilm reactor was significantly increased by 3.8-fold (2208 IU/mL) when using the best complex medium tested.