In situ and non-cytotoxic cross-linking strategy for 3D printable biomaterials.

3D bioprinting allows the production of patient-specific tissue constructs with desired structural characteristics such as high resolution, controlled swelling degree, and controlled degradation behavior by mostly using hydrogels. Crosslinking of hydrogels is an essential parameter in bioprinting applications, which is beneficial for tuning structural specifications. In this study, gelatin-alginate-whey protein isolate based hydrogels have been used for 3D printing structures in a layer-by-layer fashion. These structures were cross-linked by the Amino Acid (monomer) Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method, which is a unique, non-invasive photosensitive cross-linking technique for protein-based mixtures. In that aim, hydrogel properties (e.g., printability, biocompatibility, rheologic and mechanical behavior) and cross-linking properties (e.g., swelling and degradation behavior) were studied. Results were compared with UV and ionic cross-linking techniques, which are the abundantly used techniques in such studies. The results showed that the ANADOLUCA method can be used for in situ cross-linking under mild conditions for the printing of bio-inks, and the proposed method can be used as an alternative for UV-based and chemical cross-linking techniques.

RuBisCO nano enzyme for mimicking CO2 conversion system in plants.

In this study, carbon dioxide (CO2 ) capture and conversion systems based on the combination of biomimetic systems with nano enzymes have been developed. The effectiveness of the developed system has been investigated toward CO2 conversion. For this aim, nano ribulose bisphosphate carboxylase/oxygenase (RuBisCO) enzyme that plays role in the Calvin cycle in photosynthesis has been synthesized in 93 nm size according to AmiNoAcid (monomer) Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method. Enzymatic activity of synthesized nano RuBisCO enzyme has been spectrophotometrically determined by the formation of 3-phosphoglycerate (3-PGA) at the end of the reaction between CO2 and d-ribulose-1,5 biphosphate with the catalysis of RuBisCO enzyme at 340 nm. The effect of substrate concentration, pH, temperature, and Mg2+ ion concentration on the conversion reaction have investigated comparatively with nano and free RuBisCO enzyme. Besides this, the reusability feature of synthesized nano RuBisCO enzyme in conversion of CO2 reaction is indicated. When all data were evaluated, it has been seen that the nano RuBisCO enzyme is effective on the conversion of CO2 into 3-PGA and can be used for CO2 capture and conversion systems repeatedly without any deformation in its structure.

A new potentiometric platform: Antibody cross-linked graphene oxide potentiometric immunosensor for clenbuterol determination.

An immunosensor is based on the signal measurement obtained upon the reaction of an antibody antigen complex. It plays a significant role in various fields such as environmental analysis, production monitoring, drug detection or screening, veterinary medicine, and animal food. In this study, an antibody crosslinked graphen oxide (GO)-based potentiometric sensor has been developed for fast, simple, and economical detection of clenbuterol. In this context, the photosensitive amino acid bound GO platform is synthesized and used for the preparation of electrode material. Then, polymeric structure is characterized by infrared spectroscopy, and the performance of immunonano platform prepared by potentiometric sensor is evaluated. The effect of pH, response time, selectivity, and sensitivity is investigated. Under the optimized conditions, a simple and rapid method for the determination of clenbuterol from milk sample is established by immuno-potentiometric sensor. The detection limit has found to be 0.87 × 10-9 mmol L-1 for this immuno-potentiometric sensor. This immuno-potentiometric sensor has optimum pH at 7.0, a wide linear response (1.0 × 10-2 to 1.0 × 10-9 mmol L-1 ), rapid response time (2 Min) and 36 weeks operational lifetime.

Proteinous Polymeric Shell Decorated Nanocrystals for the Recognition of Immunoglobulin M.

This study demonstrates the preparation of photosensitively orientated and crosslinked proteinous polymeric shell having quantum dot based nanocrystals through Amino acid Decorated and Light Underpinning Conjugation Approach (ANADOLUCA). ANADOLUCA is based on photo-electron transfer method and uses these decorated nanocrystals for specifically and effectively recognition and detection of Immunoglobulin M in the aqueous environment. The conjugation method effectively provides an orientation of affinity pairs on the surface of quantum dots nanocrystals. This photosensitive ruthenium-based amino acid monomer is a synthetic and inexpensive material for the preparation of bioconjugates. The nanocrystals give advantages for using a wide pH and temperature range. The construction and preparation method is applicable to silica materials, superparamagnetic particles, quantum dots, carbon nanotubes, Ag/Au nanoparticles, Au surfaces, and polymeric materials. This prepared proteinous polymeric shell decorated nanocrystals are of great potential in applications in life sciences and can be used in infection case studies or allergy symptoms.

Molecular Imprinting Technology in Quartz Crystal Microbalance (QCM) Sensors.

Molecularly imprinted polymers (MIPs) as artificial antibodies have received considerable scientific attention in the past years in the field of (bio)sensors since they have unique features that distinguish them from natural antibodies such as robustness, multiple binding sites, low cost, facile preparation and high stability under extreme operation conditions (higher pH and temperature values, etc.). On the other hand, the Quartz Crystal Microbalance (QCM) is an analytical tool based on the measurement of small mass changes on the sensor surface. QCM sensors are practical and convenient monitoring tools because of their specificity, sensitivity, high accuracy, stability and reproducibility. QCM devices are highly suitable for converting the recognition process achieved using MIP-based memories into a sensor signal. Therefore, the combination of a QCM and MIPs as synthetic receptors enhances the sensitivity through MIP process-based multiplexed binding sites using size, 3D-shape and chemical function having molecular memories of the prepared sensor system toward the target compound to be detected. This review aims to highlight and summarize the recent progress and studies in the field of (bio)sensor systems based on QCMs combined with molecular imprinting technology.

Developing column material for the separation of serum amyloid P and C reactive protein from biological sources.

In this study, we have investigated the isolation of serum amyloid P (SAP) and C-reactive protein (CRP) from rainbow trout. It has recently been found that SAP is deposited in atherosclerotic lesions or neurofibrillary tangles, which are related to aging process and Alzheimer's disease. Given the importance of CRP, the CRP level in blood is becoming recognized as a potential means of monitoring cardiovascular risk. These two proteins, members of the pentraxin family of oligomeric serum proteins, were isolated from rainbow trout using N-methacryloyl-phosphoserine (MA-pSer) immobilized poly (2-hydroxy ethylmethacrylate) (PHEMA) cryogels as a column material in a fast protein liquid chromatography system. The separation process was verified in two steps. First, SAP and CRP proteins were isolated together from serum sample of rainbow trout using MA-pSer/PHEMA cryogel columns. Second, SAP protein was separated chromatographically from CRP protein using the Ca(2+) ion immobilized PHEMA cryogel column. According to the data, a new and effective technique has been developed for the isolation of SAP and CRP proteins from a biological source, rainbow trout. Finally, purified SAP and CRP were loaded using sodium dodecyl sulfate-polyacrylamide gel and western blot analysis to investigate the purity of chromatographically isolated SAP and CRP compared with commertial SAP and CRP.

A comprehensive assessment of redox balance in small for gestational age newborns and their mothers.

OBJECTIVE: The objective of this study was to assess oxidative stress in small for gestational age (SGA) newborns and their mothers by evaluating intra- and extracellular thiol homeostasis and the quantification of major oxidants and antioxidants. METHODS: A total of 75 mothers and their 75 newborns (43 SGA) were enrolled in this study. Thiol-disulfide homeostasis, serum myeloperoxidase, catalase, total oxidant, and antioxidant status were analyzed. Additionally, erythrocytic glutathione (GSH) homeostasis was measured. RESULTS: Although native and total thiol levels were decreased, disulfide levels were increased in SGA groups. Additionally, myeloperoxidase activity and total oxidant status levels were significantly elevated whereas total antioxidant status levels and enzymatic antioxidant systems were diminished in SGA groups. Similarly, intra-erythrocytic GSH homeostasis was shifted in favor of oxidants in SGA groups. CONCLUSION: Our results demonstrate that insufficient antioxidant systems in mothers and a robust source of oxidative stress in SGA might contribute to the pathophysiology of SGA births.

New synthesis method for 4-MAPBA monomer and using for the recognition of IgM and mannose with MIP-based QCM sensors.

Quartz crystal microbalance (QCM) sensors coated with molecularly imprinted polymers (MIP) have been developed for the recognition of immunoglobulin M (IgM) and mannose. In this method, methacryloylamidophenylboronic acid (MAPBA) was used as a monomer and mannose was used as a template. For this purpose, initially, QCM electrodes were modified with 2-propene-1-thiol to form mannose-binding regions on the QCM sensor surface. In the second step, the methacryloylamidophenylboronic acid-mannose [MAPBA-mannose], pre-organized monomer system, was prepared using the MAPBA monomer. Then, a molecularly imprinted film was coated on to the QCM electrode surface under UV light using ethylene glycol dimethacrylate (EDMA), and azobisisobutyronitrile (AIBN) as a cross-linking agent and an initiator, respectively. The mannose can be simultaneously bound to MAPBA and fitted into the shape-selective cavities. The binding affinity of the mannose-imprinted sensors was investigated using the Langmuir isotherm. The mannose-imprinted QCM electrodes have shown homogeneous binding sites for mannose (K(a): 3.3 × 10(4) M(-1)) and heterogeneous binding sites for IgM (K(a1): 1.0 × 10(4) M(-1); K(a2): 3.3 × 10(3) M(-1)).

Bioconjugated and cross-linked bionanostructures for bifunctional immunohistochemical labeling.

The present study describes the development and use of a new bioconjugate combining targeted quantum dot labeling with an immunoperoxidase method and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1). The new bioconjugate is designed for the examination of samples both under fluorescent and bright-field microscopy at the same time. For this purpose chlorobis(2-2'-bipyridyl) methacryloyl tyrosine-ruthenium(II) and bis (2-2'-bipyridyl) methacryloyltyrosine-methacryloyltryptophan-ruthenium (II) photosensitive monomers and photosensitive poly(Bis (2-2'-bipyridyl)) methacryloyltyrosine-methacryloyltryptophan-ruthenium(II) were synthesized and characterized. The anti-SOD1 antibody and horseradish peroxidase (HRD) conjugated quantum dots were prepared by using this polymer. The anti-SOD1 antibody and HRD conjugated quantum dots were used in labeling and imaging of SOD1 in rat liver sections. Quantum dot particles were observed as a bright fluorescence in their specific binding locations inside the hepatocytes. The HRD-diaminobenzidine reaction product was observed as brown-colored particles at the same locations under bright-field microscopy. Structural details of the tissue sections could be examined at the same time. The conjugation protocol is simple; the bioconjugate is applicable for efficient cell labeling and can be adapted for imaging of other targets in different tissues. Also, the prepared nanobioconjugates have mechanic stability and can be used for a long period.

Application of HRP-streptavidin bionanoparticles for potentiometric biotin determination.

Biotin is widely used in infant formula to prevent biotin deficiency of newborn babies and in beauty products as nutritional supplements for coenzymatic functions and having strong nails, shiny hair, and skin over the last few years. There is a need for the development of a fast, simple and reusable assay method to perform biotin determination at very low concentrations. Biotin determination has achieved with a prepared potentiometric biotin sensor that has a very wide concentration range (10-15M-10-7M) and a lower detection limit (0.3 10-15M) with a very good regression coefficient (0.9925). A quick response (7 min), good accuracy (recovery 100.4-103.7%), reproducible, reusable (10 times), and long-term stability (3 months) have been obtained using the prepared potentiometric sensor. The obtained results have proved that the prepared potentiometric sensor can be used for biotin determination in real samples.

Investigation of photosensitively bioconjugated targeted quantum dots for the labeling of Cu/Zn superoxide dismutase in fixed cells and tissue sections.

This study presents the development of targeted and antibody cross-linked QDs and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1) on fixed cells and tissues. QD-antibody conjugation was achieved by using our previously invented AmiNoacid (monomer) Decorated and Light Underpining Conjugation Approach (ANADOLUCA) method. In this method, we have used a photosensitive aminoacid monomer having ruthenium complex which is a synthetic and inexpensive material for the preparation of bioconjugates. Its specificity was demonstrated by extracting the active enzyme from rat liver lysate by using the bioconjugate. It provided accurate antibody orientation, high specificity and mechanic stability. The protocol steps for QD-antibody conjugation and specimen preparation were described in detail. The nanobioconjugates were prepared under mild conditions (for example in day light), independent of pH and temperature, without affecting conformation and function of protein. This protocol is simple, inexpensive and can be successfully adapted to detect other targets on different cell types and tissues.

Graphenoxide Cross-Linker Based Potentiometric Biosensor Design for Sarcosine Determination.

BACKGROUND: Sarcosine, also known as N-methyl glycine, is a natural amino acid that is an intermediate and by product in glycine synthesis and degradation. Recently found in many peptides, sarcosine has been researched as a newly accepted prostate cancer marker. The increased concentration of sarcosine in blood serum and the urine showed that malignancy of measured prostate cancer cells is active. OBJECTIVE: In this article, we aimed to design a potentiometric biosensor for detection of sarcosine with a low detection limit, high selectivity, short response time, wide linear range, and satisfactory long-term stability. METHODS: In this article, we developed a new Graphene oxide (GFOX) photosensitive cross-linker based potentiometric biosensor based on the AmiNoAcid (monomer) Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method. The functional groups determined using Raman, FT-IR, XPS analyzes, and surface characterization, the morphology of synthesized GFOX photosensitive cross-linker were determined by TEM and AFM studies. Then, the performance of the GFOX based potentiometric biosensor has been evaluated. RESULTS: When the usage of the developed GFOX doped potentiometric biosensor against sarcosine determination, it was found that 10-4 mM sarcosine was determined in 60 seconds in the solution. In addition, the detection limit of the GFOX doped potentiometric biosensor was found to be 9.45x10-7 mM, and the linear potentiometric biosensor was found to be in the concentration range of 10-1 to 10-5 mM. The selectivity studies of the developed potentiometric biosensor were investigated using glycine solutions, and it was determined that GFOX doped potentiometric biosensor was more selective against sarcosine. Besides this, a reusability test using 10-3 mM sarcosine solution showed that reproducible studies were performed without the loss of potential of designed potentiometric biosensor and no loss of sensitivity. CONCLUSION: After applying the framework, we get a new potentiometric biosensor for sarcosine determination. GFOX photosensitive cross-linker was used in designing potentiometric biosensors, and this increased the stability and efficiency of the biosensor. Therefore, the developed potentiometric biosensor for sarcosine determination could be easily used for the early diagnosis of prostate cancer.

Preconcentration of phosphate ion onto ion-imprinted polymer.

In this study, selective separation and preconcentration of phosphate ions on the phosphate-imprinted chitosan-succinate beads have investigated. Chitosan-succinate, phosphate, epichlorohydrin were used as the complexing monomer, template and crosslinking agent, respectively. In the first step, chitosan was modified with succinic anhydrides and complex formation occurred between carboxylic acid functional groups and iron(III) ions. Secondly, Fe(III)-chitosan-succinate particles were reacted with phosphate ions. Afterwards, particles were crosslinked with epichlorohydrin and the template (phosphate ions) was removed using 1M KOH solution. Selective cavity for the phosphate ion was obtained in the phosphate-imprinted metal-chelate polymer. These phosphate-imprinted metal-chelate polymer was used in the adsorption-desorption process. The adsorption process was fast and equilibrium was reached around 30 min. The adsorption behaviour of this system was described approximately by the Langmuir equation. Percent extraction, distribution ratio and selectivity coefficients of phosphate and other ions using non-imprinted and phosphate-imprinted polymer were also determined and comparison of these data was reported.

Removal of mercury species with dithiocarbamate-anchored polymer/organosmectite composites.

Mercury is one of the most toxic heavy metals found in solid and liquid waste disposed by chloro-alkali, paint, paper/pulp, battery, pharmaceutical, oil refinery and mining companies. Any form of mercury introduced to nature through any means is converted into a more toxic form such as methylmercury chloride (as produced by aquatic organisms) which usually accumulates in the tissue of fish and birds. The primary aim of this study was to investigate performance of dithiocarbamate-anchored polymer/organosmectite composites as sorbents for removal of mercury from aqueous solution. The modified smectite nanocomposites then were reacted with carbondisulfide to incorporate dithiocarbamate functional groups into the nanolayer of the organoclay. These dithiocarbamate-anchored composites were used for the removal of mercury species [Hg(II), CH(3)Hg(I) and C(6)H(5)Hg(I)]. Mercury adsorption was found to be dependent on the solution pH, mercury concentration and the type of mercury species to be adsorbed. The maximum adsorption capacities were equal to 157.3 mg g(-1) (782.5 micromol g(-1)) for Hg(II); 214.6 mg g(-1) (993.9 micromol g(-1)) for CH(3)Hg(I); 90.3 mg g(-1) (325 micromol g(-1)) for C(6)H(5)Hg(I). The competitive adsorption capacities (i.e. adsorption capacities based on solutions containing all three mercuric ions) are 7.7 mg g(-l) (38.3 micromol g(-1)), 9.2 mg g(-l) (42.6 micromol g(-1)) and 12.7 mg g(-1) (45.7 micromol g(-1)) for Hg(II), CH(3)Hg(I) and C(6)H(5)Hg(I), respectively, at 10 ppm initial concentration. The adsorption capacities on molar basis were in order of C(6)H(5)Hg(I)>CH(3)Hg(I)>Hg(II).

Nano-hemoglobin film based sextet state biomemory device by cross-linked photosensitive hapten monomer.

In this study, a biomemory device, consisting of hemoglobin (Hb) cross-linked by MACys-Ru(bipyr)2-MACys) photosensitive monomer cross-linkers, which have memory effect through both Ru3+/2+ in hapten monomer and Fe3+/2+ in redox active center of Hb through multi-charge transfer mechanism, has been improved. Cyclic voltammetry (CV) has been used to determine the redox property of the Hb cross-linked MACys-Ru(bipyr)2-MACys) hapten. Three memory functions, writing, reading and erasing of the fabricated biomemory device, have been accomplished by chronoamperometry (CA) and open-circuit potential amperometry (OCPA). The reliability and repeatability of the biodevice consisting of the p(Hb-co-MACys-Ru(bipyr)2-MACys) sextet state bio-memory layer have been analysed. The Hb film based biodevice on gold electrodes has shown ≥ 2 months the retention time and switched until 106 times continuous cycling without degradation in efficiency. Other hand, the topography of p(Hb-co-MACys-Ru(bipyr)2-MACys) layer on the gold surface has investigated by scanning electron microscopy (SEM) and EDX data.

Ferritin based bionanocages as novel biomemory device concept.

Ferritin is an iron cage having protein, capable of extracting metal ions in their cages and a consequence of the electron transfer of metal ions in their cage by reduction and oxidation processes, electrochemical information storage devices can be designed. In this work, ferritin based protein biomemory substrate has been synthesized by using Amino Acid (monomer) Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method, which utilizes photosensitive electron transfer based microemulsion co-polymerization as nanobead form of ferritin. Protein substrate contains metal ions such as silver and copper or metal ion pairs namely, silver-copper (Janus bionanocage) and co-polymeric shell of the photosensitive crosslinker protein. The redox behavior of bionanocages differentiates electrochemical "writing" and "erase" states depending on these metal ions (silver or copper) or metal ion pairs. The bionanocages based biomemory substrates have been immobilized using graphene modified glassy carbon electrodes and the memory functions of ferritin based bionanocages have been confirmed by chronoamperometry (CA) and open circuit potential amperometry (OCPA). The stability and durability of multi-state memory devices represent promising properties for future bioelectronic information technologies.

3D Micropatterned All-Flexible Microfluidic Platform for Microwave-Assisted Flow Organic Synthesis.

A large-area, all-flexible, microwaveable polydimethoxysilane microfluidic reactor was fabricated by using a 3D printing system. The sacrificial microchannels were printed on polydimethoxysilane substrates by a direct ink writing method using water-soluble Pluronic F-127 ink and then encapsulated between polydimethoxysilane layers. The structure of micron-sized channels was analyzed by optical and electron microscopy techniques. The fabricated flexible microfluidic reactors were utilized for the acetylation of different amines under microwave irradiation to obtain acetamides in shorter reaction times and good yields by flow organic synthesis.

Ion-imprinted beads for molecular recognition based mercury removal from human serum.

The aim of this study is to prepare ion-imprinted polymers which can be used for the selective removal of mercury ions [Hg(2+)] from human serum. N-Methacryloyl-(L)-cysteine (MAC) was chosen as the complexing monomer. In the first step, Hg(2+) was complexed with MAC and the Hg(2+)-imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-cysteine) (MIP) beads were synthesized by suspension polymerization. After that, the template ions (i.e., Hg(2+)) were removed using thiourea (0.5%, v/v) in 0.05 M HCl. The specific surface area of the MIP beads was found to be 59.04 m(2)/g with a size range of 63-140 micro m in diameter and the swelling ratio was 91.5%. According to the elemental analysis results, the MIP beads contained 87.0 micro mol MAC/g polymer. The maximum adsorption capacity was 0.45 mg Hg(2+)/g beads. The applicability of two kinetic models including pseudo-first order and pseudo-second order model was estimated on the basis of comparative analysis of the corresponding rate parameters, equilibrium capacity and correlation coefficients. Results suggest that chemisorption processes could be the rate-limiting step in the adsorption process. The relative selectivity coefficients of MIP beads for Hg(2+)/Cd(2+), Hg(2+)/Zn(2+) were 14.7 and 21.5 times greater than the non-imprinted (NIP) matrix, respectively. The MIP beads could be used many times without decreasing in their adsorption capacities significantly.

Cr(III)-imprinted polymeric beads: Sorption and preconcentration studies.

The purpose of this study is to prepare a novel molecular imprinted adsorbent to Cr(III) analysis with high selectivity. Initially, chromium (III)-methacryloylhistidine (MAH/Cr(III)) complex monomer was synthesized and then Cr(III) ion imprinted ethyleneglycoldimethacrylate-methacryloylhistidine (poly(EDMA-MAH/Cr(III))) was polymerized. Cr(III) ions were removed from the Cr(III)-imprinting polymer. Selective cavity for the chromium (III) ions was obtained in the (poly(EDMA-MAH/Cr(III))) beads. These Cr(III)-imprinted beads were used in the sorption-desorption process. The effect of initial concentration of metal, the sorption rate and the pH of the medium on sorption capacity of Cr(III)-imprinting sorbents were studied. Sorption equilibrium time was achieved in about 30min. The maximum sorption of Cr(III) ions onto imprinted beads was about 69.28mg/g. Sorption studies of Co(II), Ni(II), Cr(III) and Cr(VI) ions were also investigated using Cr(III)-imprinting polymers. The obtained results show that selectively adsorbed amount of Cr(III) ion on Cr(III)-imprinting polymers is higher than all other studied ions. When the beads were used repeatedly, their adsorption capacity was showed feasibility.

Phosphoserine imprinted nanosensor for detection of Cancer Antigen 125.

Cancer antigen 125 (CA 125) is widely used as diagnostic biomarker for ovarian cancer. Change in the concentration level of CA 125 is associated with disease progression or regression. CA 125 posseses a phosphorylation site and protein backbone is phosphorylated on serine, before secretion. In this study, we have developed an imprinting method for CA 125 recognition and determination. In this method, methacryloyl antipyrine europium (III) [(MAAP)2-Eu(III)] and methacryloyl antipyrine terbium (III) [(MAAP)2-Tb(III)] have been used as new metal-chelating monomers via metal coordination-chelation interactions. Phosphoserine (PS) has been used as a template for the detection of CA 125. PS imprinted carbon nanotube (CNT) and Fe2O3 nanoparticle (SPN) have cavities that are selective for CA 125. The binding affinity of the PS imprinted CNT and SPN nanosensor has been investigated using Langmuir adsorption isotherms and affinity constants (Kaffinity) have found to be 1.85 105M-1 for PS and 13.5 10-3 mLU-1 and 7.73 10-3 mLU-1 for CA 125 (for CNT and SPN, respectively). Detection limit of PS imprinted CNT nanosensor for PS and CA 125 have been found to be 1.77 10-10M and 0.49 UmL-1, respectively. Human serum samples have been spiked with different concentrations of CA 125 (in pH 7.4 PBS) to investigate the feasibility of the nanosensors for clinical applications. Experimental results have been revealed that prepared nanosensors have been exhibited better sensivity, recovery and reproducibility.