Fluorimetric Reusable Polymeric Sensor for Hydrogen Sulfide Detection.

In this study, with the help of reactive monomers, crosslinkers, and photoinitiator that detect H2S in various matrices, an H2S sensitive fluorescence sensor polymerizes under ultraviolet (UV) light was developed. To this goal, a polymeric membrane was prepared, and the characterization of the membrane was carried out with Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) methods. Afterward, appropriate conditions were identified, the excitation wavelength was determined as 370 nm, and the emission wavelength was determined as 425 nm. It was established that the fluorescence intensity of the prepared polymeric membrane decreased in the presence of H2S. A detailed analysis was executed to determine the sensor's most suitable pH value and time. It was found that the optimum pH was 8.0, and the optimal duration was 15 s. It has been calculated that the linear range of the developed method is 2.19 × 10-8- 6.25 × 10-7 M, and the detection limit (LOD) is 7.37 × 10-9 M. The effect of some possible interfering ions was investigated, and it determined that the sensor had excellent selectivity. In addition, the sensor used to determine H2S can be used at least 100 times. The recovery percentages were 102.1%-103.2%, and 104.6%, using tap water samples. In terms of providing reliable, fast results, high sensitivity, reusable, low cost, and ease of use, the developed fluorimetric sensor, compared to standard methods, has become more advantageous.

Molecularly imprinted nanoparticles with recognition properties towards diphtheria toxin for ELISA applications.

Plastic antibodies can be used for in vitro neutralization of biomacromolecules with different fragments due to their potential in separation, purification, chemical sensor, catalysis and drug production studies. These polymer nanoparticles with binding affinity and selectivity comparable to natural antibodies were prepared using functional monomer synthesis and copolymerization of acrylic monomers via miniemulsion polymerization. As a result, the in vitro cytotoxic effect from diphtheria toxin was reduced by MIPs. In vitro imaging experiments of polymer nanoparticles (plastic antibodies) were performed to examine the interaction of diphtheria toxin with actin filaments, and MIPs inhibited diphtheria toxin damage on actin filaments. The enzyme-linked immunosorbent assay (ELISA) was performed with plastic antibodies labeled with biotin, and it was determined that plastic antibodies could also be used for diagnostic purposes. We report that molecularly imprinted polymers (MIPs), which are biocompatible polymer nanoparticles, can capture and reduce the effect of diphtheria toxic and its fragment A.

Macromolecules can be imprinted by using their fragments as template molecules.MIPs gain an affinity for the template molecule by covalent binding, non-covalent interactions or ligand interactions, as well as the ability to bind, release and recognize the template molecule.The viability of cells treated with DT, NIPs and MIPs was determined by MTT assay.Immunofluorescence staining studies examined structural changes in actin filaments in HUVEC treated with DT, NIPs and MIPs.FA imprinted polymer has the ability to bind whole diphtheria toxin.FA-MIP gave significant results in terms of specificity in ELISA using diphtheria toxin.

An intelligent indoor guidance and navigation system for the visually impaired.

Intelligent guidance in complex environments where various procedures are required for navigation is critical to achieving mobility for the visually impaired. This study presents a newly developed software prototype with a hybrid RFID/BLE infrastructure to provide intelligent navigation and guidance to the visually impaired in complex indoor environments. The system enables the users to input their purpose via a specially designed user interface, and provides intelligent guidance through a chain of destination targets which are determined according to the inherent procedures of the environment. Path optimization is performed by adaptation of the traveling salesman problem, and real-time instantaneous instructions are provided to guide the users through the predetermined destination points. For evaluation purposes, a hospital environment is constructed as an example of a complex environment and the system is tested by visually impaired participants. The results show that the intelligent purpose selection and destination evaluation mechanism modules of the system are found to be effective by all the participants.

A new perspective on the pleiotropic blood pressure improvement effect of sitagliptin: downregulation of miRNA-155 and miRNA-21.

INTRODUCTION: Deterioration of vascular responses is the crucial event in the initiation of cardiovascular problems in hypertension (HT) and diabetes mellitus (DM). A well-known oral antidiabetic, sitagliptin, has pleiotropic effects besides improving glycemic state in type-2 DM. This study aimed to investigate the therapeutic effect of sitagliptin on blood pressure with previously unassessed parameters of well-known pathophysiological processes and especially at the microRNA (miRNA) level where there are many unknowns. METHODS: N-nitro-L-arginine methyl ester (L-NAME)-induced HT model was performed on nondiabetic male rats. Four groups (including 7 rats in each) were formed: normotensives, sitagliptin-treated, HT and sitagliptin-treated HT. Asymmetric dimethylarginine (ADMA), intercellular adhesion molecule-1 (ICAM-1) and tyrosine hydroxylase (TH), HT related miRNAs were evaluated. In-vitro vessel responses were observed. RESULTS: L-NAME led to a significant increase in blood pressure. Hypertensives exhibited significantly increased contractile responses, consistent with increased ADMA, ICAM-1. Sitagliptin decreased TH levels but not statistically significantly. The new side of the study was the miRNA-21 and miRNA-155 expressions were in line with other parameters in both the HT and sitagliptin-treated HT groups. CONCLUSION: Sitagliptin may control comorbidities, especially HT and introduces new targets to alleviate vascular responses. The new knowledge is; sitagliptin may show these effects through microRNAs (Tab. 2, Fig. 6, Ref. 46).

Abnormal neovascular and proliferative conjunctival phenotype in limbal stem cell deficiency is associated with altered microRNA and gene expression modulated by PAX6 mutational status in congenital aniridia.

PURPOSE: To evaluate conjunctival cell microRNA (miRNAs) and mRNA expression in relation to observed phenotype of progressive limbal stem cell deficiency in a cohort of subjects with congenital aniridia with known genetic status. METHODS: Using impression cytology, bulbar conjunctival cells were sampled from 20 subjects with congenital aniridia and 20 age and sex-matched healthy control subjects. RNA was extracted and miRNA and mRNA analyses were performed using microarrays. Results were related to severity of keratopathy and genetic cause of aniridia. RESULTS: Of 2549 miRNAs, 21 were differentially expressed in aniridia relative to controls (fold change ≤ -1.5 or ≥ +1.5). Among these miR-204-5p, an inhibitor of corneal neovascularization, was downregulated 26.8-fold in severely vascularized corneas. At the mRNA level, 539 transcripts were differentially expressed (fold change ≤ -2 or ≥ +2), among these FOSB and FOS were upregulated 17.5 and 9.7-fold respectively, and JUN by 2.9-fold, all being components of the AP-1 transcription factor complex. Pathway analysis revealed enrichment of PI3K-Akt, MAPK, and Ras signaling pathways in aniridia. For several miRNAs and transcripts regulating retinoic acid metabolism, expression levels correlated with keratopathy severity and genetic status. CONCLUSION: Strong dysregulation of key factors at the miRNA and mRNA level suggests that the conjunctiva in aniridia is abnormally maintained in a pro-angiogenic and proliferative state, and these changes are expressed in a PAX6 mutation-dependent manner. Additionally, retinoic acid metabolism is disrupted in severe, but not mild forms of the limbal stem cell deficiency in aniridia.

Effects of cage type on performance, welfare, and microbiological properties of laying hens during the molting period and the second production cycle.

As most of the unenriched cages will soon switch to enriched cages, it is important to characterize all the effects in the laying hens for sustainable production. Laying hens can be used in several production periods by applying molting. The aim of this study was to determine the cage type (unenriched and enriched) on performance, welfare, and microbiological properties of laying hens during the molting period and the second production cycle. Overall, 840 brown laying hybrids were used in the experiment. Laying hens were reared on two different cage types (unenriched cage (UEC) and enriched cage (EC)) in the same poultry house. When the hybrids were 75 weeks old, they were subjected to force molting with whole grain barley. Performance, welfare, microbiological, and serological data of laying hens were obtained from 73 to 107 weeks of age. Egg production, egg weight, feed conversion ratio, breaking strength, albumen and yolk index, Haugh unit, feather condition, and breaking force of femur and metatarsus were better in the post-molting period. However, keel bone deformities and Newcastle disease virus antibody titers are the worst in the post-molting period. Stiffness of femur and metatarsus was increased with period. These results indicate that necessary precautions should be taken against the problems that may occur in the direction of bone and health. During the molting period, hens kept in EC had lower egg production but they returned to egg production at a high rate. EC type had a positive effect on egg production, feed conversion ratio, feather and foot condition, and breaking force of metatarsus.

Polymeric nanoparticles for selective protein recognition by using thiol-ene miniemulsion photopolymerization.

The fabrication of molecularly imprinted nanoparticles (MIP-NPs) specific for myoglobin by using thiol-ene photopolymerization in miniemulsion was described. Allyl derivatives of phenylalanine as a functional monomer was synthesized and copolymerized with acrylic monomers via miniemulsion polymerization to produce NIP-NPs with approximately 74 nm number average particle diameter. FTIR and 1H-NMR analysis confirmed the synthesis of functional monomer. MIP-NPs were prepared in the existence of myoglobin as a template protein. Morphological investigations exhibited that the particle size of the MIP-NPs, increased compared to the corresponding NIPs and the mean particle diameter by number was measured as 141 nm with narrow distribution. NIP-NPs that were polymerized without myoglobin were found to have less affinity to the target protein. In addition, the rebinding ability of MIP-NPs was much bigger than that of the corresponding NIPs. ELISA results showed that MIPs interact particularly with the myoglobin and show little affinity for BSA in competitive binding experiments.HighlightsAllyl N,N-diallyl phenylalaninate was synthesized as a functional monomer.Imprinted nanoparticles were prepared by using thiol-ene photopolymerization in miniemulsion.The nanoparticles were 141 nm with narrow size distribution.The imprinted nanoparticles showed selectivity toward myoglobin.

A New Fluorescent Sensor for Arsenic(III) Determination in Aqueous Media.

A novel polymeric membrane sensor was developed by using 2-hydroxyethyl acrylate, trimethylolpropane triacrylate, (3-mercaptopropyl)trimethoxysilane, and poly(ethylene glycol)diacrylate for As(III) determination. Various parameters, like the pH, response time, the foreign ions, and concentration effects were investigated for deciding the optimum working conditions of the polymeric sensor. As a result of this investigation, the optimum pH was found to be 2, and the response time was found to be 30 s. The linear range of the sensor was 6.65 × 10-9 - 3.99 × 10-8 mol L-1 (0.50 - 2.99 µg L-1) with a detection limit of 2.33 × 10-9 mol L-1 (0.18 µg L-1). Soy flour and well-water samples were successfully analyzed with the developed sensor. The sensor can be used at least 100 times after regeneration. It could be reused by washing with purified water and showed good stability for 6 months.

Limited Long-Term Impact of Insect Venom Immunotherapy on the Micro-RNA Landscape in Whole Blood.

OBJECTIVE: To perform a genome-wide characterization of changes in microRNA (miRNA) expression during the course of venom immunotherapy (VIT). METHODS: miRNA was isolated from the whole-blood of 13 allergic patients and 14 controls, who experienced no allergic reaction upon stings by honeybees and wasps. We analyzed 2549 miRNAs from the whole blood of these patients prior to VIT and 12 months after the start of VIT. The results for differential expression obtained on a microarray platform were confirmed by quantitative real-time PCR. Out of the 13 patients, 8 had a negative allergic reaction with VIT, thus indicating that this approach was successful. RESULTS: By comparing time points before and 12 months after ultrarush VIT, correlation analysis and principal component analysis both indicated a limited effect of VIT on the overall miRNA expression pattern. Volcano plot analysis based on raw P values revealed few deregulated miRNAs, most of which were increasingly expressed after VIT as compared with before VIT. Based on the 50 most altered miRNAs, no clear clustering was observed before or after VIT. CONCLUSIONS: Our results indicate an overall reduced effect of VIT on the miRNA expression pattern in whole blood.

Covalent immobilization of acetylcholinesterase on a novel polyacrylic acid-based nanofiber membrane.

In this study, polyacrylic acid-based nanofiber (NF) membrane was prepared via electrospinning method. Acetylcholinesterase (AChE) from Electrophorus electricus was covalently immobilized onto polyacrylic acid-based NF membrane by demonstrating efficient enzyme immobilization, and immobilization capacity of polymer membranes was found to be 0.4 mg/g. The novel NF membrane was synthesized via thermally activated surface reconstruction, and activation with carbonyldiimidazole upon electrospinning. The morphology of the polyacrylic acid-based membrane was investigated by scanning electron microscopy, Fourier Transform Infrared Spectroscopy, and thermogravimetric analysis. The effect of temperature and pH on enzyme activity was investigated and maxima activities for free and immobilized enzyme were observed at 30 and 35°C, and pH 7.4 and 8.0, respectively. The effect of 1 mM Mn2+, Ni2+, Cu2+, Zn2+, Mg2+, Ca2+ ions on the stability of the immobilized AChE was also investigated. According to the Michaelis-Menten plot, AChE possessed a lower affinity to acetylthiocholine iodide after immobilization, and the Michaelis-Menten constant of immobilized and free AChE were found to be 0.5008 and 0.4733 mM, respectively. The immobilized AChE demonstrated satisfactory reusability, and even after 10 consecutive activity assay runs, AChE maintained ca. 87% of its initial activity. Free enzyme lost its activity completely within 60 days, while the immobilized enzyme retained approximately 70% of the initial activity under the same storage time. The favorable reusability of immobilized AChE enables the support to be employable to develop the AChE-based biosensors.

Differences in egg nutrient availability and embryo development in white layer breeder genotypes.

Because of consumers' preferences and also due to changes in production systems, the importance of pure breeds has increased again. There are a lot of differences among breeds which have been studied extensively, however, the differences during the incubation period are not yet fully known. Therefore, the present study was conducted to evaluate the composition of the egg parts, absorption of nutrients, and development of embryos from different genotypes. A total of 354 fresh hatching eggs were obtained from one hybrid (Lohman White, LW) and two pure breeds (Denizli and Gerze). Hatching eggs from each genotype were examined on the day of setting for egg analysis and then at the beginning of the embryonic d 19 (E19) and embryonic d 21 (E21) for egg, embryo, jejunum, and tibia analysis. On d 21 of incubation, the healthy chicks were removed and weighed. Egg weight, shell thickness, percentages of albumen, and some parameters of albumen composition (dry matter, water, ash, protein, energy, Na, Ca, K, and Mg) were higher in fresh eggs obtained from LW hens. Furthermore, the relative yolk sac and embryo weight, some yolk parameters (dry matter, water, protein, fat, and energy) and some shell parameters (dry matter, ash, Na, Ca, and K) were also higher in eggs obtained from LW hens during incubation. However, tibia deformation and villus width were lower in LW embryos than the other genotypes. Relative chick weights were 68.9, 72.0, and 68.0% in LW, Denizli, and Gerze genotypes, respectively. During incubation, differences in all examined parameters were significant except thickness and weight of shell, tibia deformation, and crypt depth. Yolk sac weight, some yolk composition parameters, K level in the shell, Cu level in the tibia, and villus height were also affected by genotype and period interaction. Based on these results, LW was found advantageous in terms of egg composition, however, regarding villus development and tibia deformation in embryos during incubation, pure breeds showed better results.

Meningoencephalitis caused by pathogenic Sarcocystis species in a naturally infected sheep in Turkey.

A 3-year-old sheep was examined after an acute onset of hind limb paralysis and ataxia. At necropsy, central nervous system, pulmonary and intestinal hyperaemia and ecchymoses in the aortic arch were observed. Main microscopic lesions were confined to the heart, cerebrum and cerebellum. There were a multifocal mild myocarditis and nonsuppurative meningoencephalitis together with protozoal cysts in the heart and the brain. Protozoal cystic structures were observed within many of the myocardial fibers as well as in the cerebrum and cerebellum. Using light microscopy it could not be morphologically determined whether these organisms were Toxoplasma (T.) gondii or Neospora (N.) caninum. Additional diagnostic methods like immunohistochemistry and polymerase chain reaction provided differentiation of Sarcocystis from T. gondii and N. caninum. Transmission electron microscopy demonstrated characteristic features of Sarcocystis sp. as previously described. This is the first confirmed diagnosis of Sarcocystis sp. in the central nervous system of a sheep from Turkey.

Effects of choline treatment in concentrations of serum matrix metalloproteinases (MMPs), MMP tissue inhibitors (TIMPs) and immunoglobulins in an experimental model of canine sepsis.

The aim of the present study was to investigate effects of intravenous (i.v.) choline treatment on serum matrix metalloproteinases (MMP), MMP tissue inhibitors (TIMP) and immunoglobulins (Igs), and to determine if there were relations between serum MMPs/TIMPs and C-reactive protein (CRP) (as a marker of the acute phase response), immunoglobulin G and M (IgG and IgM) (as a maker of the Ig responses) and markers of organ damage such as muscular damage (creatine phosphokinase, [CPK]), liver damage (alanine aminotransferase [ALT]) and renal dysfunction (blood urea nitrogen [BUN] and creatinine, [Cr]) in dogs with endotoxemia. Healthy dogs (n=24) were randomized to Saline, Choline (C), Lipopolysaccharide (LPS), and LPS+C groups and received 0.9% NaCl (5mL/i.v.), choline chloride (20mg/kg/i.v.), LPS (0.02mg/kg/i.v.) and LPS (0.02mg/kg/i.v.) plus choline chloride (20mg/kg/i.v.), respectively. Serum MMPs and TIMPs concentrations were analyzed by commercial ELISA kits. MMP and TIMP increased at 1-48h (P<0.05), whereas IgG and IgM decreased at 24-48h in LPS group, compared to their baselines. Choline treatment reduced changes in serum MMPs, TIMPs and markers of organ damage, and prevented the hypoimmunoglobulinemia in LPS+C. MMPs and TIMPs were correlated positively (P<0.05) with serum CRP, CPK, ALT, BUN and Cr, but not with serum Igs. Our findings suggest that the serum MMPs, TIMPs and Igs are involved in the pathophysiology of endotoxemia, and MMPs and TIMPs are correlated with the acute phase reaction and multi-organ failure. In addition, we demonstrated a direct effect of choline administration in decreasing serum MMPs and TIMPs, and preserving serum Igs in the course of endotoxemia.

Matrix metalloproteinase expression in primary lung fibroblasts of layer type chickens.

We attempted to determine the growth characteristics of cultured lung fibroblasts of layer type chickens and to investigate presence of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in these cells in vitro. Lung fibroblasts were isolated, characterized and subcultured from one-day-old layer type chicken lungs. Two different methods, explant culture and enzymatic techniques, were used for culturing and the results were compared. The presence of MMP-2 and TIMP-1 was shown in cultured fibroblasts by immunocytochemical staining, immune blotting and zymography methods. Immune expressions of neither MMP-9 nor TIMP-2 enzymes could be detected.

Preparation of biocompatible, UV-cured fumarated poly(ether-ester)-based tissue-engineering hydrogels.

The aim of this study was to develop biodegradable, photo-polymerizable in situ gel-forming systems prepared from a fumaric acid monoethyl ester (FAME) modified poly(lactide-co-glycolide) (PLGA) co-polymer. By reacting lactide and glycolide in the presence of stannous octoate as a catalyst and 2-ethyl,2-hydroxymethyl 1,3-propanediol as an initiator, hydroxyl terminated branched PLGA was synthesized. Afterwards, at room temperature hydroxyl terminated branched PLGA was reacted with fumaric acid monoethyl ester (FAME). N,N'-dicyclohexylcarbodiimide and triethylamine were used as a coupling agent and catalyst, respectively. The gel percentage, equilibrium mass swelling, degradation profile and polymerization kinetics of the hydrogels were investigated. All of the results were influenced by the amount of FAME modified PLGA co-polymer. Biocompatibility of the hydrogels was examined by using MTT cytotoxicity assay. According to the results, hydrogels are biocompatible and cell viability percentage depends on the amount of PLGA co-polymer. While the amount was 15% in hydrogel composition, cell viability was 100%, but after increasing the PLGA co-polymer amount to 30% the viability reduced to 78%.

Slurry analysis of cadmium and copper collected on 11-mercaptoundecanoic acid modified TiO2 core-Au shell nanoparticles by flame atomic absorption spectrometry.

Separation/preconcentration of copper and cadmium using TiO(2) core-Au shell nanoparticles modified with 11-mercaptoundecanoic acid and their slurry analysis by flame atomic absorption spectrometry were described. For this purpose, at first, titanium dioxide nanoparticles were coated with gold shell by reducing the chloroauric acid with sodium borohydride and then modified with 11-mercaptoundecanoic acid. The characterization of modified nanoparticles was performed using ultra-violet spectroscopy and dynamic light scattering. Copper and cadmium were then collected on the prepared sorbent by batch method. The solid phase loaded with the analytes was separated by centrifugation and the supernatant was removed. Finally, the precipitate was slurried and directly aspirated into the flame for the determination of analytes. Thus, elution step and its all drawbacks were eliminated. The effects of pH, amount of sorbent, slurry volume, sample volume and diverse ions on the recovery were investigated. After optimization of experimental parameters, the analytes in different certified reference materials and spiked water samples were quantitatively recovered with 5% RSD. The analytes were enriched up to 20-fold. Limits of detection (N=10, 3σ) for copper and cadmium were 0.28 and 0.15 ng mL(-1), respectively.

Photopolymerized injectable RGD-modified fumarated poly(ethylene glycol) diglycidyl ether hydrogels for cell growth.

In this study, photopolymerized hydrogels of fumarated poly(ethylene glycol) diglycidyl-co- poly(ethylene glycol) diacrylate have been synthesized and modified with cell adhesion peptide, Arg-Gly-Asp (RGD). The structural and mechanical properties of the hydrogels are found to be poly(ethylene glycol) diacrylate (PEGDA) dependent. The percentage of gelation is increased from 72 to 89 wt.-% when the amount of the crosslinker co-monomer (PEGDA) in the hydrogel formulation is increased from 20 to 40 wt.-%. In the present case, the equilibrium mass swelling is decreased from 216 to 93%. The viscosities of the uncured formulations have also been measured and likewise, the results were influenced by the increasing amount of PEGDA that reduced the value from 83 to 36 cP. The compressive modulus of the prepared hydrogels was improved with the addition of the PEGDA. Cell growth experiments have been performed by comparing the properties of the hydrogels with and without RGD units. The results show that RGD units enhance the adhesion of cells to the surface of the hydrogels. SEM-EDS studies reveal that nitrogen and calcium are produced on the osteoblast-seeded surface of the scaffold within the culture time period. [Figure: see text].

Targeted surveillance to assess the presence of BSE in the age risk population of cattle slaughtered in Bursa, Turkey: preliminary results of an immunohistochemical detection study for the 2004-2005 period.

Bovine spongiform encephalopathy (BSE), a member of the transmissible spongiform encepahlopathies, has been a notifiable disease in Turkey since 1997. In 2002, the BSE status of Turkey was assessed by the EU Scientific Steering Committee as "it is likely but not confirmed". This study presents the results of a targeted surveillance study to assess the presence of BSE in the age risk population of Bursa, Turkey. In the assessment procedure, the immunohistochemical detection of protease-resistant prion protein (PrP-Sc) was aimed at and applied to 420 brain tissues of cattle slaughtered in Bursa at an age of 30-months and older. None of the samples were positive for BSE.

The identification of matrix metalloproteinases and their tissue inhibitors in broiler chickens by immunohistochemistry.

The aim of this study was to investigate the distribution of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and TIMP-2 using immunohistochemistry in the ascites syndrome of broiler chickens in a salt-induced experimental model. The presence of the enzymes in the lung, heart, liver, kidney and brain was evaluated semi-quantitatively with the streptavidin-biotin-peroxidase (Strep-ABC) method using commercially available primary monoclonal antibodies. Immunostaining of MMP-2 and MMP-9 was more intense and extensive in ascitic broilers than in the controls, although a decrease was seen with increasing age both in normal and ascitic chickens. The presence of MMP-9 enzyme was negatively correlated with the presence of TIMP-1 enzyme. It is suggested that MMP-2 and MMP-9 enzymes might play a role in the permeability increase of vessel walls by the destruction of the basement membranes in the salt-induced experimental ascites syndrome in broiler chickens.