3D incoherent imaging using an ensemble of sparse self-rotating beams.

Interferenceless coded aperture correlation holography (I-COACH) is one of the simplest incoherent holography techniques. In I-COACH, the light from an object is modulated by a coded mask, and the resulting intensity distribution is recorded. The 3D image of the object is reconstructed by processing the object intensity distribution with the pre-recorded 3D point spread intensity distributions. The first version of I-COACH was implemented using a scattering phase mask, which makes its implementation challenging in light-sensitive experiments. The I-COACH technique gradually evolved with the advancement in the engineering of coded phase masks that retain randomness but improve the concentration of light in smaller areas in the image sensor. In this direction, I-COACH was demonstrated using weakly scattered intensity patterns, dot patterns and recently using accelerating Airy patterns, and the case with accelerating Airy patterns exhibited the highest SNR. In this study, we propose and demonstrate I-COACH with an ensemble of self-rotating beams. Unlike accelerating Airy beams, self-rotating beams exhibit a better energy concentration. In the case of self-rotating beams, the uniqueness of the intensity distributions with depth is attributed to the rotation of the intensity pattern as opposed to the shifts of the Airy patterns, making the intensity distribution stable along depths. A significant improvement in SNR was observed in optical experiments.

Pullout strength of misplaced pedicle screws in the thoracic and lumbar vertebrae - A cadaveric study.

BACKGROUND: The objective of this cadaveric study was to analyze the effects of iatrogenic pedicle perforations from screw misplacement on the mean pullout strength of lower thoracic and lumbar pedicle screws. We also investigated the effect of bone mineral density (BMD), diameter of pedicle screws, and the region of spine on the pullout strength of pedicle screws. MATERIALS AND METHODS: Sixty fresh human cadaveric vertebrae (D10-L2) were harvested. Dual-energy X-ray absorptiometry (DEXA) scan of vertebrae was done for BMD. Titanium pedicle screws of different diameters (5.2 and 6.2 mm) were inserted in the thoracic and lumbar segments after dividing the specimens into three groups: a) standard pedicle screw (no cortical perforation); b) screw with medial cortical perforation; and c) screw with lateral cortical perforation. Finally, pullout load of pedicle screws was recorded using INSTRON Universal Testing Machine. RESULTS: Compared with standard placement, medially misplaced screws had 9.4% greater mean pullout strength and laterally misplaced screws had 47.3% lesser mean pullout strength. The pullout strength of the 6.2 mm pedicle screws was 33% greater than that of the 5.2 mm pedicle screws. The pullout load of pedicle screws in lumbar vertebra was 13.9% greater than that in the thoracic vertebra (P = 0.105), but it was not statistically significant. There was no significant difference between pullout loads of vertebra with different BMD (P = 0.901). CONCLUSION: The mean pullout strength was less with lateral misplaced pedicle screws while medial misplaced pedicle screw had more pullout strength. The pullout load of 6.2 mm screws was greater than that of 5.2 mm pedicle screws. No significant correlation was found between bone mineral densities and the pullout strength of vertebra. Similarly, the pullout load of screw placed in thoracic and lumbar vertebrae was not significantly different.

Antibodies against high frequency Gerbich 2 antigen (anti-Ge2): A real challenge in cross matching lab.

Transfusion management of patients' alloimmunized against high-prevalence erythrocyte antigens is often problematic in emergency situations. Gerbich (Ge) is very common blood group system and Gerbich-2 (Ge-2) antigen present in high frequency and outside Papua New Guinea population, Ge-2 negative population almost nil. To manage such kind of problems with real emergencies, implementation of rare donor registry program, cryopreservation of red cells of rare donors and biological cross matching to assess significance of these antibodies is warranted.

Nanofabrication of bio-self assembled monolayer and its electrochemical property for toxicant detection.

Cyclic voltammetry (CV) has been used to investigate the electrochemical behavior of a glutathione (GSH) self assembled monolayer on modified gold electrodes (Bio-SAM). The GSH monolayer exhibits an influence on electrode surface activity. Electrochemically immobilized dsDNA onto a Cyt c/GSH-SAM/Au electrode, which is useful for the fabrication of a nanobiosensing device. The immobilized Cyt c followed by dsDNA immobilized films maintained its surface activity and finally dsDNA/Cyt c/GSH-SAM/Au electrode, targeted for the detection of toxicants. The films were characterized by CV, DPV, and AFM. The differential pulse voltammetry (DPV) technique was applied to detect three kinds of common toxins, 2-aminoanthracene (2-AA), 3-bromobenzanthrone (3-BBA) and bisphenol A (BPhA). The electrochemical signals showed good inverse relationship with the increase of concentrations of toxicants. Our proposed system based on electrochemical method with nanoscale film technology can be applied at highly sensitive biosensor for detecting various toxic chemicals.

A catechol biosensor based on a gold nanoparticles encapsulated-dendrimer.

Tyrosinase has been immobilized on a Au nanoparticles encapsulated-dendrimer bonded conducting polymer on a glassy carbon electrode for the estimation of catechol. The modified electrode was characterized by cyclic voltammetry and AFM techniques. The principle of catechol estimation was based on the reduction of biocatalytically liberated quinone species at +0.2 V versus Ag/AgCl (3 M KCl), with good stability, sensitivity, and featuring a low detection limit (about 0.002 µM) and wide linear range (0.005 µM-120 µM). The electrochemical redox peak of catechol on the GCE/PolyPATT/Den(AuNPs)/tyrosinase was also investigated. A response time of 7 s, reusability up to 5 cycles and a shelf life of more than 2 months under refrigerated conditions were reported. Various parameters influencing biosensor performance have been optimized including pH, temperature, and applied potential. The utility and application of this nanobiosensor was tested in a real water samples.

nanosieve using : a sensor for detection of.

A novel approach for the preparation of a well-ordered nanosieve structure of silver nanoparticles using a Ag-1,2-benzenedicarboxylate (Ag-1,2-BDC) complex and sodium stearate under a hydrothermal process was performed. The X-ray diffraction (XRD) pattern suggests the formation and crystallinity of silver nanoparticles. The average particle size of silver nanoparticles was 35 nm and shows a hexagonal shape with a nanosieve structure as confirmed by transmission electron microscopy (TEM). The selective area electron diffraction (SAED) pattern of the nanosieve suggests a mono-crystalline nature of the silver nanoparticles. The SEM image reveals that the particles are of spherical and granular nature. The UV-Vis absorption spectrum shows a characteristic absorption peak of silver nanoparticles at 410 nm. The findings suggest that hydrothermal induced assembly should be a convenient and effective route for the preparation of new silver nanostructures. In addition, the Ag nanosieve modified electrode (AgNS/GC) exhibited an excellent electrocatalytic activity toward the reduction of hydrogen peroxide (H2O2). This silver nanosieve may also be a promising candidate for electronics and photonics application.

Application of peptide nucleic acid towards development of nanobiosensor arrays.

Peptide nucleic acid (PNA) is the modified DNA or DNA analogue with a neutral peptide backbone instead of a negatively charged sugar phosphate. PNA exhibits chemical stability, resistant to enzymatic degradation inside living cell, recognizing specific sequences of nucleic acid, formation of stable hybrid complexes like PNA/DNA/PNA triplex, strand invasion, extraordinary thermal stability and ionic strength, and unique hybridization relative to nucleic acids. These unique physicobiochemical properties of PNA enable a new mode of detection, which is a faster and more reliable analytical process and finds applications in the molecular diagnostics and pharmaceutical fields. Besides, a variety of unique characteristic features, PNAs replace DNA as a probe for biomolecular tool in the molecular genetic diagnostics, cytogenetics, and various pharmaceutical potentials as well as for the development of sensors/arrays/chips and many more investigation purposes. This review paper discusses the various current aspects related with PNAs, making a new hot device in the commercial applications like nanobiosensor arrays.

Quality assessment of platelet concentrates prepared by platelet rich plasma-platelet concentrate, buffy coat poor-platelet concentrate (BC-PC) and apheresis-PC methods.

BACKGROUND: Platelet rich plasma-platelet concentrate (PRP-PC), buffy coat poor-platelet concentrate (BC-PC), and apheresis-PC were prepared and their quality parameters were assessed. STUDY DESIGN: IN THIS STUDY, THE FOLLOWING PLATELET PRODUCTS WERE PREPARED: from random donor platelets (i) platelet rich plasma - platelet concentrate (PRP-PC), and (ii) buffy coat poor-platelet concentrate (BC-PC) and (iii) single donor platelets (apheresis-PC) by different methods. Their quality was assessed using the following parameters: swirling, volume of the platelet concentrate, platelet count, WBC count and pH. RESULTS: A total of 146 platelet concentrates (64 of PRP-PC, 62 of BC-PC and 20 of apheresis-PC) were enrolled in this study. The mean volume of PRP-PC, BC-PC and apheresis-PC was 62.30+/-22.68 ml, 68.81+/-22.95 ml and 214.05+/-9.91 ml and ranged from 22-135 ml, 32-133 ml and 200-251 ml respectively. The mean platelet count of PRP-PC, BC-PC and apheresis-PC was 7.6+/-2.97 x 1010/unit, 7.3+/-2.98 x 1010/unit and 4.13+/-1.32 x 1011/unit and ranged from 3.2 -16.2 x 1010/unit, 0.6-16.4 x 1010/unit and 1.22-8.9 x 1011/unit respectively. The mean WBC count in PRP-PC (n = 10), BC-PC (n = 10) and apheresis-PC (n = 6) units was 4.05+/-0.48 x 107/unit, 2.08+/-0.39 x 107/unit and 4.8+/-0.8 x 106/unit and ranged from 3.4 -4.77 x 107/unit, 1.6-2.7 x 107/unit and 3.2 - 5.2 x 106/unit respectively. A total of 26 units were analyzed for pH changes. Out of these units, 10 each were PRP-PC and BC-PC and 6 units were apheresis-PC. Their mean pH was 6.7+/-0.26 (mean+/-SD) and ranged from 6.5 - 7.0 and no difference was observed among all three types of platelet concentrate. CONCLUSION: PRP-PC and BC-PC units were comparable in terms of swirling, platelet count per unit and pH. As expected, we found WBC contamination to be less in BC-PC than PRP-PC units. Variation in volume was more in BC-PC than PRP-PC units and this suggests that further standardization is required for preparation of BC-PC. As compared to the above two platelet concentrates, all the units of apheresis-PC fulfilled the desired quality control criteria of volume. Apheresis-PC units showed better swirling and platelet count than PRP-PCs and BC-PCs. All the platelet concentrates units had pH well above the recommended norm.

Direct immobilization of cupredoxin azurin modified by site-directed mutagenesis on gold surface.

For making efficient bioelectronic device, we have developed novel immobilization method of cupredoxin azurin modified on gold (Au) surface. A recombinant protein with cysteine residue by using site-directed mutagenesis was designed and then directly immobilized on Au surface without any chemical linker. The immobilization of the functionalized protein is confirmed by surface plasmon resonance (SPR) and its surface morphology is analyzed by scanning tunneling microscopy (STM). The immobilization efficiency has been increased about 75.6%, as compared to that of wild-type azurin. The electrochemical property of the fabricated thin film was investigated by the cyclic voltammetry (CV). As a result, cysteine-modified azurin can be used for making high-quality protein film, and applied to the fabrication of nano-scale bioelectronics.

Analysis of direct immobilized recombinant protein G on a gold surface.

For the immobilization of IgG, various techniques such as chemical linker, thiolated protein G methods, and fragmentation of antibodies have been reported [Y.M. Bae, B.K. Oh, W. Lee, W.H. Lee, J.W. Choi, Biosensors Bioelectron. 21 (2005) 103; W. Lee, B.K. Oh, W.H. Lee, J.W. Choi, Colloids Surf. B-Biointerfaces, 40 (2005) 143; A.A. Karyakin, G.V. Presnova, M.Y. Rubtsova, A.M. Egorov, Anal. Chem. 72 (2000) 3805]. Here, we modified the immunoglobulin Fc-binding B-domain of protein G to contain two cysteine residues at its C-terminus by a genetic engineering technique. The resulting recombinant protein, RPGcys, retained IgG-binding activity in the same manner as native protein G. RPGcys was immobilized on a gold surface by strong affinity between thiol of cysteine and gold. The orientations of both IgG layers immobilized on the base recombinant protein Gs were analyzed by fluorescence microscope, atomic force microscope (AFM), and surface plasmon resonance (SPR). Our data revealed that IgG-binding activity of RPGcys on gold surface significantly increased in comparison to wild type of protein G (RPGwild), which was physically adsorbed due to absence of cysteine residue. Immobilization of highly oriented antibodies based on cysteine-modified protein G could be useful for the fabrication of immunosensor systems.

Application of electrochemically prepared polypyrrole-polyvinyl sulphonate films to DNA biosensor.

Double stranded calf thymus deoxyribonucleic acid (DNA) was physisorbed onto polypyrrole-polyvinyl sulphonate (PPY-PVS) films electrochemically deposited onto indium-tin-oxide (ITO) coated glass plates. These DNA immobilized PPY-PVS films optimized for various conditions, such as polymerization potential, pH of buffer, DNA concentration and scan rate were characterized using Fourier-transform infrared (FT-IR) spectroscopy, atomic force microscopy (AFM) and cyclic voltammetry (CV) techniques, respectively. The amperometric response studies of these DNA/PPY-PVS electrodes were carried out as a function of 2-aminoantharcene (2-AA, 0.01-20 ppm) and o-chlorophenol (OCP, 0.1-30 ppm) concentration, respectively at 25 degrees C. The observed amperometric current arising due to oxidation of guanine in the DNA/PPY-PVS films decreased linearly with the increase in the concentration of 2-AA and OCP. It has been revealed that 10 ppm of 2-AA is sufficient to reduce the observed guanine oxidation peak current by approximately -95+/-10% as compared to the reported values. A 25 ppm of OCP was capable enough to reduce the guanine oxidation current to zero. These DNA/PPY-PVS electrodes were found to have a shelf life of about 4 months when stored at 25 degrees C.

Application of octadecanethiol self-assembled monolayer to cholesterol biosensor based on surface plasmon resonance technique.

Octadecanethiol (ODT) self-assembled monolayer (SAM) prepared onto gold-coated glass plate has been modified by using nitrene reaction of 1-fluoro-2-nitro-4-azidobenzene (FNAB) that further covalently binds to cholesterol oxidase (ChOx) via thermal reaction. FNAB acts as a bridge (cross-linker) between SAM and ChOx. The ChOx/FNAB/ODT/Au electrode thus fabricated has been characterized using contact angle (CA) measurements, UV-vis spectroscopy, electrochemical techniques and X-ray photoelectron spectroscopy (XPS) technique, respectively. This ChOx/FNAB/ODT/Au bioelectrode has been utilized for estimation of cholesterol in solution using surface plasmon resonance (SPR) technique. This SPR based cholesterol biosensor has linearity from 50 to 500mg/dl of cholesterol in solution with lower detection limit of 50mg/dl and shelf life of about 2 months when stored at 4 degrees C.

Enhancement of urinary elimination of 3-bromobenzanthrone metabolites by oral supplementation of ascorbic acid in guinea pigs.

OBJECTIVE: 3-Bromobenzanthrone (3-BBA), an anthraquinone intermediate dye, is extensively used in textile industry. Since, our prior studies have shown that 3-BBA caused significant depletion of ascorbic acid (AsA) levels, the effect of exogenous supplementation of AsA on the urinary elimination of 3-BBA metabolites was investigated. METHOD: Guinea pigs were treated with single oral dose of 3-BBA (50 mg/kg b. wt.) in groundnut oil while another group was treated with single oral dose of 3-BBA (50 mg/kg b. wt.) along with 3 day prior and post oral supplementation of AsA. Control groups were either treated with groundnut oil or AsA alone. Urine from individual animals was collected, extracted and analysed on HPTLC. RESULTS: The highest elimination of 3-BBA (75 microg) was found to be in 0-24 h urine fraction which decreased to 18 microg and 5 microg in the two subsequent 24 hourly fractions of urine. Exogenous supplementation of AsA increased the total urinary elimination of 3-BBA by almost 77%. A total of 10 fluorescent metabolites excluding the parent compound were eliminated in the urine of guinea pigs treated with 3-BBA. Densitometric scanning of chromatogram showed different peaks at Rf 0.18, 0.22, 0.27, 0.34, 0.40, 0.48, 0.56, 0.66, 0.72, 0.80, and 0.95 which were eliminated and marked as urinary metabolite 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 respectively. AsA not only significantly enhanced the elimination of 3-BBA metabolites but also modified the pattern of metabolites drastically in 0-6 h, 6-24 h and 24-48 h urine fractions. CONCLUSION: These results indicate that AsA may be useful in protecting the toxicity of 3-BBA by fascilitating the urinary metabolite(s) excretion of 3-BBA.

Bio-elimination of conjugated metabolites of 3-bromobenzanthrone in urine of rats and Guinea pigs.

The profile of urinary metabolites of 3-bromobenzanthrone (3-BBA), an extensively used anthraquinone dye intermediate, in rats and guinea pigs was investigated using HPTLC system. A total of 10 fluorescent metabolites were detected in the urine of guinea pigs as compared to 8 in rats including the parent compound 3-BBA. The elimination of metabolites increased in a dose dependent manner in rats. The Rf values of metabolites in rats were 0.14, 0.29, 0.42, 0.52, 0.58, 0.64, 0.77 and 0.91 while that in guinea pigs were 0.23, 0.26, 0.34, 0.37, 0.44, 0.54, 0.56, 0.68, 0.76 and 0.95. The urine of 3-BBA (50 mg/kg b.wt) treated guinea pigs when digested with acid showed the disappearance of metabolite 1, 2, 5 and 7 indicating these to be the conjugated metabolites. Further, digestion of urine of 3-BBA treated guinea pigs with glucuronidase showed disappearance of metabolite 5 and 7 suggesting these as glucuronide conjugates. Digestion of urine with sulfatase enzyme resulted in disappearance of metabolite 1 which could be a sulfate conjugate. Urinary metabolite 2 which was found to be present even after digestion with glucuronidase or sulfatase but disappeared following acid treatment appears to be glutathione conjugate(s) which resulted in formation of metabolite 3 and 6. These results suggest that conjugated fluorescent metabolites of 3-BBA are excreted in urine of rats and guinea pigs.

Noncanonical vortex transformation and propagation in a two-dimensional optical system.

We produced an axial and canonical optical vortex by using a computer-generated hologram and then converted it to a noncanonical vortex by passing it through a cylindrical lens. We conducted an experimental study of the shape and trajectory of the noncanonical vortex as it propagates in free space and obtained an analytical expression explaining our experimental results. The computed trajectory and shape of the noncanonical vortex agree quite well with our experimental results.

Comparative effect of benzanthrone and 3-bromobenzanthrone on hepatic xenobiotic metabolism and anti-oxidative defense system in guinea pigs.

Benzanthrone (BA) and 3-bromobenzanthrone (3-BBA) are important dye intermediates used in the manufacture of various vat and disperse dyes. BA has been implicated as a cause of hepatic malfunctions and dermal lesions in workers. However, not much information on halogenated BAs, especially 3-BBA, is available. Experiments were designed to undertake a comparative safety assessment of both BA and 3-BBA, given orally at a dose of 50 mg/kg body weight for 10 days to guinea pigs. There was a significant decrease (25%) in body weight with 3-BBA, whereas BA treatment did not cause any change. Serum glutamate oxaloacetate transaminase and glutamate pyruvate transminase were found to be significantly (P<0.05) increased in 3-BBA- as well as in BA-treated animals. 3-BBA and BA led to substantial depletion of ascorbic acid in both liver and adrenal glands. However, depletion of ascorbic acid was more pronounced with 3-BBA (19.2-28.3%) than with BA (13.5-16.6%). 3-BBA and BA treatments caused 80% and 24% depletion of hepatic free sulfydryl content, while lipid peroxidation showed a significant enhancement of 73% and 47%, respectively. BA and 3-BBA caused decreases in cytochrome P-450 content and phase I enzymes particularly ethoxyresorufin- O-deethylase and aryl hydrocarbon hydroxylase, whereas phase II enzymes (quinone reductase and glutathione- S-transferase) were substantially increased. Activities of bio-antioxidant enzymes, viz., glutathione peroxidase, glutathione reductase, superoxide dismutase and catalase, were significantly increased by 153, 104, 20 and 67% in the 3-BBA-treated group, whereas the degree of increase in these parameters was relatively less in BA-treated group. The data indicate that both BA and 3-BBA can disturb membrane integrity by decreasing endogenous glutathione and ascorbic acid levels with a concomitant increase in lipid peroxidative damage. This may in turn lead to impairment of hepatic P-450-dependent monooxygenase, while the changes in antioxidant enzymes reveal oxidative stress. 3-BBA treatment caused dilation of portal triad with thickening of arterial wall, hyperplasia of Kupffer cells and influx of inflammatory cells between hepatic cords, which could be due to formation of Br(*) radical or due to formation of semiquinone type of intermediate following oxidation. The results may be interpreted to mean that industrial workers exposed to 3-BBA are at higher risk than those exposed to BA, and necessary precautions should be taken to safeguard their exposure risks.

A sensitive method of monitoring exposure to 3-bromobenzanthrone in industrial dyestuff workers.

3-Bromobenzanthrone (3-BBA) is an anthraquinone dye intermediate widely used for the synthesis of a variety of dyes. The monitoring of 3-BBA exposure in dyestuff industry workers has not been possible until now as no procedure has been available. In this article, the fluorescence properties of 3-BBA has been utilized to develop a quantitative method for the detection of this dye intermediate. The procedure allows the measurement of trace quantities of 3-BBA in biological specimens, including urine, serum, liver, and feces, in experimental studies. The method involves extraction of biological samples with an equal volume of a chloroform-methanol mixture (1:1, v/v) followed by measurement of the relative fluorescence intensity at excitation maxima of 400 nm and emission maxima of 530 nm. The detection limit was found to be as low as 50 ng of 3-BBA. The procedure can be routinely used to screen for the presence of 3-BBA in biological fluids, especially urine, as a measure of exposure to 3-BBA in dyestuff workers. The authors are thankful to Dr. P. K. Seth, Director, Industrial Toxicology Research Centre, for his keen interest in this study. The secretarial assistance of Mr. K. G. Thomas is duly acknowledged. RPS is thankful to University Grant Commission, New Delhi, for the award of a Senior Research Fellowship.