Effect of Ankaferd Blood Stopper on muscle healing.

PURPOSE: Ankaferd Blood Stopper (ABS), which is a standardized mixture of herbal extracts obtained from five plants, has been proven as an efficient hemostatic agent and is still used in emergency situations. It is not known exactly if decreased bleeding has positive or negative effects on muscle healing and fibrosis, so the purpose of this study was to test the effect of ABS on muscle healing and morphology. METHODS: A total of 66 outbred Wistar rats were divided into three control and three experimental subgroups. In the experimental groups, ABS was sprayed on the cut surface of the soleus. In the control groups, a saline solution was sprayed on the cut surface of the soleus. Subgroups were euthanized after 2 weeks, 3 weeks and 4 weeks, respectively. In each subgroup, eight rats were used for the biomechanical study to determine muscle healing and three rats were used for the histopathological investigation. RESULTS: Although muscle strength in the control groups was lower than that of the experimental groups in early weeks, no differences were found between the control and the experimental groups at 4 weeks. CONCLUSIONS: ABS has no negative effect on muscle healing. We also observed that ABS accelerated muscle healing compared to the control group. ABS could be used in hemostasis of open fractures and elective orthopedic surgeries.

An alternative treatment method for defective pseudoarthrosis; evaluation of eight patients treated with Artelon+Kryptonite.

Pseudoarthrosis with bone loss is one of the most challenging orthopaedic problems for surgeons. Bone loss usually leads to technical difficulties during surgery due to instability in the fracture area. Eight patients with pseudoarthrosis of different long bones were operated on by the same surgeon. The median age was 53 years (25-61), and the median time period after the index operation was 21 months (12-72 months). Radiographic union was achieved in all patients in 3.62 months (2-5 months). Efficient healing with new bone formation was observed in all of the patients. The result of the current case series is promising. This treatment method can be used for the treatment of pseudoarthrosis without increasing morbidity. Long-term follow-up and larger case series are needed for evidence of the adequacy and reliability of this method of treatment.

Micromotor-Based Biomimetic Carbon Dioxide Sequestration: Towards Mobile Microscrubbers.

We describe a mobile CO2 scrubbing platform that offers a greatly accelerated biomimetic sequestration based on a self-propelled carbonic anhydrase (CA) functionalized micromotor. The CO2 hydration capability of CA is coupled with the rapid movement of catalytic micromotors, and along with the corresponding fluid dynamics, results in a highly efficient mobile CO2 scrubbing microsystem. The continuous movement of CA and enhanced mass transport of the CO2 substrate lead to significant improvements in the sequestration efficiency and speed over stationary immobilized or free CA platforms. This system is a promising approach to rapid and enhanced CO2 sequestration platforms for addressing growing concerns over the buildup of greenhouse gas.

Determination of carcinoembryonic antigen (CEA) by label-free electrochemical immunosensor using functionalized boron nitride nanosheets.

In this study, a simple, specific and sensitive immunosensor for CEA detection was prepared based on BN nanosheets. Lewis acid-base interaction was sufficient for the immobilization of anti-CEA used as an antibody on the electrode surface without an activation agent. This immunosensor could be used for CEA determination without the need to use any label or secondary antibody. With its epedance-based recognition mechanism, this immunosensor offered a low LOD value of 0.017 ng/mL and a wide measurement range of 0.1-500 ng/mL and could be used for a long time. The analytical performance of this immunosensor is higher than the biosensors prepared in the literature. Compared to commercially available kits, it is attractive because it is cheap, simple and analyzes in a short time. This immunosensor, which has high selectivity against CEA in the presence of competitive agents in commercial human serum, has a high potential for clinical applications.

Immobilization of Urokinase onto Magnetically Directed Micromotors.

In this presented work, a new micromotor was prepared for urokinase immobilization. A covalent bond was constructed between the urokinase and the carboxyl groups of the graphene oxide, which is located at the outer layer of micromotors by EDC/NHS chemistry. The inner nickel layer gave magnetic properties to the micromotors and enables them to be separated from the reaction medium with the help of a simple magnet. For promising in vivo applications in the future, these micromotors do not require any fuel for their movement. The structures of the synthesized micromotors were illuminated by SEM and EDX analysis, and the movements of the micromotors were observed under an optical microscope with camera equipment. The immobilization yield of urokinase was found to be 68.07% (0.073 mg/100 µL micromotor solution) using the Bradford protein assay. In addition, to compare the activities of the immobilized and free enzymes, Lineweaver-Burk plots were constructed, and the kinetic parameters were calculated. Km values for free urokinase and immobilized urokinase were 2.0964 mM and 0.5602 mM, respectively. The maximum velocities of free and immobilized urokinase were found to be 25.25 µmol/min and 30.12 µmol/min, respectively. Also, storage stability profiles of the immobilized and free urokinase were monitored for 40-day incubation at + 4 °C, and the immobilized enzyme has 88% of its initial activity, while the free urokinase demonstrated only 30% of its initial activity. As a result, the experiments were carried out in human commercial serum, and specific activity values for free urokinase and immobilized urokinase were found to be 38.06 and 169.84 µmolmg-1 min-1, respectively.

p(HEMA)-RR241 hydrogel membranes with micron network for IgG depletion in proteomic studies.

Serum proteins can generally be considered a good source for the illness' indication and are precious resources to detect diseases such as inflammation, cancer, diabetes, malnutrition, cardiovascular diseases, Alzheimer's, other autoimmune diseases, and infections. However, one of the biggest difficulties for proteomic studies is that the majority of serum protein mass consists of only a few proteins. Albumin and Immunoglobulin (IgG) constitute 80% of total serum protein. In this study, dye ligand affinity-based hydrogel membranes were proposed as new materials with micron mesh structures. Micron mesh p(HEMA) hydrogel membranes were synthesized by using the UV-photopolymerization method, then modified with Reactive Red 241 (RR241) dye ligand to increase the affinity towards IgG. Characterizations of synthesized micron mesh p(HEMA)-RR241 hydrogel membranes were also performed. It was demonstrated by the characterization studies that; the dye was successfully incorporated into the membrane structure with the amount of 119.38 mg/g. The hydrophilic property of the hydrogel membrane was demonstrated by swelling tests and the swelling value of dye modified membrane was found to be 8 times higher than that of the plain membrane. Micron network structure, as well as the porosity, were demonstrated with SEM/ESEM studies. Optimization of IgG adsorption conditions was also studied at different parameters (pH, temperature, ion strength, initial IgG concentration). Optimum pH, temperature, and ionic strength were found to be 6.5, 25 °C, 0.05 M, respectively, and the maximum IgG absorption value was 10.27 mg/g. Finally, it was shown that the proposed materials can be used repeatedly by 5 adsorption-desorption cycles.

Peroxidase driven micromotors for dynamic bioremediation.

Phenolics are size products present in tons concentrations in industrial wastewater that can cause adverse health effects when released in the environment. As such, there is a growing interest in the development of efficient strategies for the removal of phenolic compounds from polluted water. Herein we describe the use of poly(3,4-ethylenedioxythiophene) (PEDOT)-Au/peroxidase micromotors as dynamic biocatalytic platforms for the removal of model phenolics (phenol, bisphenol A, guaiacol, pyrogallol and catechol). Micromotors are synthetized by using a simplified template electrodeposition protocol followed by covalent enzyme immobilization in the inner Au layer. Kinetic parameters revealed that enzyme immobilization in the inner micromotor layer increased over 2-fold the enzymatic activity, along with increasing operational pH and thermal stabilities. The micromotors can propel at speed of up to 60 µm/s, generating an enhanced fluid mixing that results in removal efficiencies of up to 60% as compared with the 27% removal when using free peroxidase under the same conditions. In addition, excellent activities of almost 100% were obtained within ten cycles of removal using the micromotors. This newly developed bioremediation strategy holds considerable promise in for its application in large scale water treatment systems and many relevant environmental processes.

Inulinase Immobilized Lectin Affinity Magnetic Nanoparticles for Inulin Hydrolysis.

In this presented paper, concanavalin A-modified cysteine-functionalized Fe3O4/Ag core/shell magnetic nanoparticles were synthesized and used as a support material for inulinase enzyme, which has been intensively used for the preparation of high-fructose syrup by hydrolyzing inulin. Inulinase adsorption capacity of Con A-functionalized Ag-coated magnetic nanoparticles was optimized by changing medium pH, temperature, and initial inulinase concentration, and maximum inulinase adsorption capacity was found to be 655.32 mg/g nanoparticle by using 1.00 mg/mL of inulinase solution in pH 3.0 buffer system at 25 °C. Finally, efficient inulin degradation capacity of the inulinase immobilized magnetic nanoparticles was demonstrated by TLC studies and released fructose amount was determined as 0.533 mg/mL only within the 5 min of hydrolysis. This newly developed hydrolysis strategy holds considerable promise to produce high-fructose syrup in many industries.

Boron nitride nanosheet modified label-free electrochemical immunosensor for cancer antigen 125 detection.

In this presented study, a new boron nitride nanosheets modified label-free electrochemical immunosensors were prepared for early detection of cancer antigen 125 (CA125). To aim for, boron nitride (BN) nanosheets were synthesized by conventional sonication-assisted method and then characterized. BN nanosheets were used for the surface modification of the working electrode of the screen-printed electrode (SPE). Anti CA125 antibody was then directly immobilized onto the electrode surface due to its natural affinity towards BN nanosheets. Modified electrodes were blocked with BSA and finally protected with Nafion. The newly synthesized label-free immunosensor demonstrated good detection properties to CA125 with a linear range of 5-100 U and a detection limit of 1.18 U/mL. The developed immunosensor also showed excellent reproducibility, selectivity, and stability profiles. Additionally, this immunosensor was successfully used for the detection of CA125 in artificial human serum samples along with the interfering agents. Also, it is expected that the prepared immunosensor should carry the good potential for point-of-care diagnosis in real cases.

Peroxidase Immobilized Cryogels for Phenolic Compounds Removal.

In this presented work, preparation of poly(AAm) cryogel, peroxidase immobilization onto the poly(AAm) cryogel, and usability of these enzyme modified cryogels for phenolic compounds removal were described. For this purpose, poly(AAm) cryogels were synthesized by using cryocopolymerization technique at sub-zero temperatures, and covalently functionalized with peroxidase enzyme by EDC/NHS chemistry. Characterization of the cryogels was carried out by FTIR, SEM, and EDX analysis. Maximum peroxidase loading onto poly(AAm) cryogel was found to be as 127.30 mg/g cryogel. Kinetic parameters of free and immobilized peroxidases were also investigated along with the stability tests. Finally, phenolic compounds removal efficiency of the peroxidase immobilized poly(AAm) cryogel was studied towards model phenolics such as phenol, bisphenol A, guaiacol, pyrogallol, and catechol; and very high phenolic removal efficiency was recorded.

Bacteria killer enzyme attached magnetic nanoparticles.

The objective of the present work was to develop immobilized lysozyme systems through adsorption on magnetic nanoparticles for potential usage in bacteria killing studies. For this, magnetic poly(HEMA-GMA) nanoparticles were prepared by surfactant free emulsion polymerization technique and functionalized with dye ligand Reactive Green 5. Synthesized magnetic nanoparticles were then characterized by FTIR, SEM, EDX and ESR studies. Particle size range of the polymers was found to be as 90-120 nm. Magnetic behavior was also demonstrated by ESR with the g value of 2.48. Maximum lysozyme loading was found to be as 1045.1 mg/g nanopolymer. Repeated usability of the magnetic nanoparticles was also studied. Immobilized form of lysozyme protected 85.85% of its initial activity at the end of the immobilization process. Bacteria killing capacity of the lysozyme immobilized magnetic nanoparticles were investigated by using Micrococcus lysodeikticus bacteria and it was demonstrated that all bacteria were successfully destroyed by the lysozyme immobilized magnetic nanoparticles within 5 min.

Quercetin adsorption with imprinted polymeric materials.

In this study, molecularly imprinted polymer membranes were synthesized for the recognition and adsorption of quercetin. For this, quercetin imprinted polymeric membranes [p(HEMA-MAH)] (Poly(2-hydroxyethyl methacrylate-co-N-methacryloly-l-histidinemethylester) were synthesized by UV polymerization technique using HEMA and MAH as monomers. Synthesized polymeric membranes were characterized with SEM, FTIR and swelling test. Characterized membranes were used for the direct adsorption of quercetin in a batch system. Quercetin adsorption conditions were optimized by using the quercetin imprinted polymeric membrane by altering the pH, temperature and initial quercetin concentration of the adsorption medium. Effect of adsorption time was also studied for up to 180 min. The optimum pH and temperature was determined between 4.0 and 45 °C. Maximum adsorbed amount of quercetin onto quercetin imprinted poly(HEMA-MAH) membrane was found to be as 299.6 mg/g membrane using the initial quercetin concentration of 2.0 mg/ml. Adsorbed quercetin was desorbed from the polymeric membranes with isopropyl alcohol with desorption yield of 98.3%. and repeated usability of the quercetin imprinted polymeric membranes was fallowed for 7 adsorption/desorption cycles. At the end of the 7th reuse, quercetin adsorption capacity of the quercetin imprinted poly(HEMA-MAH) membranes decreased only about 10%.

Antibody separation using lectin modified poly(HEMA-EDMA) hydrogel membranes.

Herein we describe the synthesis of Concanavalin A-poly(2-hydroxyethyl methacrylate-ethylene dimethacrylate) hydrogel membranes (via photopolymerization technique) for antibody separation from aqueous solutions. Different characterization techniques including Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy, Elemental Analysis and swelling tests revealed the highly rough morphology and spherical shape of the synthetized membranes. Attached amount of IMEO (salinization agent) onto polymeric structure and Con A binding capacity were found to be 10.85 mol/g and 3.52 mg/g, respectively. Optimum conditions for IgG adsorption such as adsorption capacity, pH and reusability profile of HMs were judiciously characterized. Maximum IgG adsorption capacity of hydrogel membrane was found to be as 26.81 mg/g. Adsorbed IgG was eluted successfully by using 2.0 M of NaCl solution. Reusability profiles of hydrogel membrane in five adsorption-desorption cycles revealed that there was no significant decrease in IgG adsorption capacity at the end of the 5th reuse. The hydrogel membranes reported here hold considerable promise as an effective sorbent system for IgG adsorption with good stability and efficient repeated usage.

Synthesis and characterization of albumin imprinted polymeric hydrogel membranes for proteomic studies.

In this presented study, a novel molecularly imprinted polymeric hydrogel membranes (PHMs) were developed to use for the albumin depletion studies. For this, albumin imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-phenylalanine methyl ester) polymeric hydrogel membranes [p(HEMA-MAP) PHMs] were synthesized by the photopolymerization technique, and then characterized by SEM, EDX, FT-IR and swelling studies. Synthesized PHMs had spherical structure and the MAP monomer incorporation onto the PHMs was determined by EDX analysis by using nitrogen stoichiometry. Also, the swelling ratio of the albumin imprinted p(HEMA-MAP) PHMs was determined as 215%. The optimum albumin adsorption condition (adsorption capacity, medium pH, adsorption rate, temperature, ionic strength) were studied and the maximum albumin adsorption capacity was found to be as 34.28 mg/g PHMs. Selectivity experiments were also carried out with the presence of the competitive proteins such as lysozyme and amylase, and the results demonstrated that the albumin imprinted p(HEMA-MAP) PHMs showed high affinity towards the BSA molecules than the competitive proteins of lysozyme and amylase. Adsorbed albumin was desorbed from the PHMs by 1.0 M of NaCl, and the reusability of the imprinted PHMs was also demonstrated for five successive adsorption-desorption cycles without any significant loss in the albumin adsorption capacity. As an application, sodium-dodecyl sulfate polyacrylamide gel electrophoresis was used to indicate the albumin depletion efficiency of albumin imprinted p(HEMA-MAP) PHMs. This presented study showed that, these imprinted membranes are promising for proteomic studies and applications, and can be used for the investigations for human diagnostics.

Ultrasound-propelled nanowire motors enhance asparaginase enzymatic activity against cancer cells.

Ultrasound-(US) propelled nanowires consisting of Au/Ni/Au/PEDOT-PPy-COOH segments are modified with asparaginase enzyme and applied as an effective anti-cancer agent. After immobilization of asparaginase onto the surface of the nanowire motors, the enzyme displays enhanced thermal and pH stabilities, improved resistance towards protease, and higher affinity for the substrate. The fast motion of the motor-carrying asparaginase leads to greatly accelerated biocatalytic depletion of asparagine and hence to a significantly enhanced inhibition efficacy against El4 lymphoma cancer cells (92%) as compared to free enzyme counterpart (17%) and other control groups. Such enhanced enzymatic activity against cancer cells is attributed to the fast motion of the motors which facilitates the interaction between the enzyme and the cancer cells. While asparaginase and EL4 tumor cells are used as a model system in the present study for cancer cell inhibition, the same mechanism can be expanded to other types of enzymes and biomolecules for the corresponding biofunctions.

Controlled release of curcumin from poly(HEMA-MAPA) membrane.

In this work, poly(HEMA-MAPA) membranes were prepared by UV-polymerization technique. These membranes were characterized by SEM, FTIR, and swelling studies. Synthesized membranes had high porous structure. These membranes were used for controlled release of curcumin which is already used as folk remedy and used as drug for some certain diseases and cancers. Curcumin release was investigated for various pHs and temperatures. Optimum drug release yield was found to be as 70% at pH 7.4 and 37 °C within 2 h period. Time-depended release of curcumin was also investigated and its slow release from the membrane demonstrated within 48 h.

DNA isolation by galactoacrylate-based nano-poly(HEMA-co-Gal-OPA) nanopolymers.

Isolation of DNA is one of the important processes for biotechnological applications such as investigation of DNA structures and functions, recombinant DNA preparations, identification of genetic factors and diagnosis and treatment of genetic disorders. The aim of this study was to synthesis and characterizes the galactoacrylate based nanopolymers with high surface area and to investigate the usability of these synthesized nanopolymers for DNA isolation studies. Nanopolymers were synthesized by the surfactant free emulsion polymerization technique by using the monomers of 2-hydroxyl ethylmethacrylate and 6-O-(2'-hydroxy-3'-acryloyloxypropyl)-1,2:3,4-di-O-isopropylidene-α-D-galactopyranose. Galactoacrylate origin of these newly synthesized nanopolymers increased the interaction between DNA and nanopolymers. Prepared nanopolymers were characterized by SEM, FT-IR and ZETA sizer analysis. Synthesized nanopolymers were spherical, and their average particle size was about 246.8 nm. Adsorption of DNA onto galactoacrylate based nanopolymers was investigated by using different pHs, temperatures, ionic strength, DNA concentrations and desorption studies and maximum DNA adsorption was found to be as 567.12 mg/g polymer at 25 °C, in pH 5.0 acetate buffer. Reusability was investigated for 5 successive reuse and DNA adsorption capacity decreased only about 10% at the end of the 5th reuse.

Preparation and characterization of silanized poly(HEMA) nanoparticles for recognition of sugars.

In this study presented, p(HEMA) nanoparticles were synthesized by the emulsion polymerization technique and then activated by a silanization agent, 3-aminopropyltriethoxysilane (APTES). The APTES-functionalized p(HEMA) nanoparticles that were synthesized were characterized by studies using the Zetasizer, FTIR and SEM. The p(HEMA)-APTES nanoparticles were further modified with phenyl boronic acid (PBA), and these boronate affinity nanoparticles were used for the recognition of some sugars such as galactose, fructose and raffinose. The system parameters (temperature and initial sugar concentration) were optimized for maximum sugar adsorption. The maximum amount of galactose, fructose, and raffinose adsorbed were found to be 4334.5 mg/g; 4334.9 and 810.0 mg/g, respectively (at 25°C, in a phosphate buffer of pH 7.0). Considering the results of this study, it can be concluded that these nanoparticles may be used as a new alternative for the specific recognition of sugar.

Nanomotors responsive to nerve-agent vapor plumes.

Enzyme-powered nanomotors responsive to the presence of nerve agents in the surrounding atmosphere are employed for remote detection of chemical vapor threats. Distinct changes in the propulsion behavior, associated with the partition of the sarin simulant diethyl chlorophosphate (DCP), offer reliable and rapid detection of the nerve-agent vapor threat.

Ni(II) chelated IDA functionalized poly(HEMA-GMA) cryogels for urease adsorption.

Cryogelic support materials have been intensively used for the purification and separation of biomolecules. Cryogels are cheap materials, they can be easily used for different purposes and their chemical and physical stabilities are very high. Cryogels can also be easily functionalized with different type of ligands and are be applicable to different affinity systems. Within these affinity systems, immobilized metal affinity chromatography (IMAC) offers efficient and simple protein purification strategies. IMAC technology has been deeply applied to bioseparations studies. In the present chapter, the preparation of a cryogel support material and the functionalization with the chelating agent iminodiacetic acid (IDA) and the subsequent Ni(II) chelation are described. Characterization techniques and the cryogel preparation method are summarized and urease adsorption studies on the metal chelate cryogel are briefly explained.