Magnetic quantum dots in biotechnology – synthesis and applications

Quantum dots (QDs) have great promise in biological imaging, and as this promise is realized, there has been increasing interest in combining the benefits of QDs with those of other materials to yield composites with multifunctional properties. One of the most common materials combined with QDs is magnetic materials, either as ions (e.g. gadolinium) or as nanoparticles (e.g. superparamagnetic iron oxide nanoparticles, SPIONs). The fluorescent property of the QDs permits visualization, whereas the magnetic property of the composite enables imaging, magnetic separation, and may even have therapeutic benefit. In this review, the synthesis of fluorescent–magnetic nanoparticles, including magnetic QDs is explored; and the applications of these materials in imaging, separations, and theranostics are discussed. As the properties of these materials continue to improve, QDs have the potential to greatly impact biological imaging, diagnostics, and treatment.     

SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM TISSUE CULTURES OF PENNISETUM AMERICANUM,AND P. AMERICANUM x P. PURPUREUM HYBRID

Immature embryos as well as explants obtained from young inflorescences of Pennisetum americanum (pearl millet) give rise to callus tissues on nutrient media containing 2,4-dichlorophenoxyacetic acid (2,4-D). A compact and pale-yellow callus that arises from the peripheral cells of the scutellum, and from the young inflorescences, undergoes further organized growth. When transferred to a 2,4-D-free medium, supplemented with indole-acetic acid or kinetin, or both, embryoids are formed in the organized areas of the callus. Embryoids show a bipolar organization with a shoot-coleorhiza (root) axis and have a coleoptile-like structure surrounded at the base by a cup-shaped structure that resembles the scutellum in texture and morphology. Embryoids show bilateral or radial symmetry and “germinate” in vitro to form plants that have been grown to maturity in soil. Similar embryogenic callus cultures have been produced from young inflorescence tissues of hybrid Pennisetum, a triploid sexually sterile hybrid of P. americanum x P. purpureum. Plants derived from these have also been transferred to soil. The regenerated plants showed normal chromosome numbers.     

Fast flow rate processes for purification of alkaline xylanase isoforms from Bacillus pumilus AJK and their biochemical characterization for industrial application purposes

This study shows the presence of five isozymic forms of alkaline xylanase from Bacillus pumilus using fast flow rate microfiltration, ultrafiltration, Q-sepharose, and phenyl sepharose chromatographic techniques. Polyacrylamide gel electrophoresis, high-performance liquid chromatography, and zymographic studies also revealed the purity of five isoforms of alkaline xylanases. Isoforms—X-I, X-III, and X-V exhibited optimum activity at pH 8.5, whereas X-II, X-IV showed maximum activity at pH 9. All isoforms were optimally active at temperature 55°C. Isoforms were found to be stable at pH 7–11, showed 92–100% residual activity after 3 hr, treatment time for most industrial applications. The isoforms retained nearly 80–86% residual activity after incubating at 45°C for 3 hr. Molecular weights of xylanase I–V, were 13.1, 15.3, 18.4, 20.1, and 21.0 kDa, respectively. Mg²⁺ ions were found to be potent activator for all isozymic forms. The Km and Vmax values of X-I, X-II, X-III, X-IV, and X-V were 6.71, 6.66, 7.14, 5.88, 6.25 mg/ml and 2,000, 1,695, 1,666.66, 1,428.57, and 1,408.45 IU/mg protein, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed the monomeric nature of all isoforms. The low-molecular masses, significantly enhanced activity in the presence of industrially suitable—low cost activator, better stability of all isoforms at pH 7–11 and at higher temperature, also presence of multiple forms of alkaline xylanase, makes this enzyme suitable for textile–paper industries. This is also the first report mentioning the purification of five isozymic forms of alkaline xylanase using fast flow rate techniques.     

The Power of Two-Dimensional Dwell-Time Analysis for Model Discrimination, Temporal Resolution, Multichannel Analysis and Level Detection

Two-dimensional (2D) dwell-time analysis of time series of single-channel patch-clamp current was improved by employing a Hinkley detector for jump detection, introducing a genetic fit algorithm, replacing maximum likelihood by a least square criterion, averaging over a field of 9 or 25 bins in the 2D plane and normalizing per measuring time, not per events. Using simulated time series for the generation of the “theoretical” 2D histograms from assumed Markov models enabled the incorporation of the measured filter response and noise. The effects of these improvements were tested with respect to the temporal resolution, accuracy of the determination of the rate constants of the Markov model, sensitivity to noise and requirement of open time and length of the time series. The 2D fit was better than the classical hidden Markov model (HMM) fit in all tested fields. The temporal resolution of the two most efficient algorithms, the 2D fit and the subsequent HMM/beta fit, enabled the determination of rate constants 10 times faster than the corner frequency of the low-pass filter. The 2D fit was much less sensitive to noise. The requirement of computing time is a problem of the 2D fit (100 times that of the HMM fit) but can now be handled by personal computers. The studies revealed a fringe benefit of 2D analysis: it can reveal the “true” single-channel current when the filter has reduced the apparent current level by averaging over undetected fast gating.     

Could DNA hydroxymethylation be crucial in influencing steroid hormone signaling in endometrial biology and endometriosis?

Endometriosis affects 10% of reproductive‐aged women. It is characterized by the growth of the endometrium, outside the uterus and is associated with infertility and chronic abdominal pain. Lack of noninvasive diagnostic tools and early screening tests results in delayed treatment and subsequently increased disease severity. Endometriosis is a disease associated with a deregulated hormonal response, therefore, understanding the molecular mechanisms that govern this hormonal interplay is of paramount importance. DNA methylation is an epigenetic mark that regulates gene expression and is often associated with genes that code for steroid receptors and enzymes associated with estrogen synthesis and metabolism in endometriosis. DNA hydroxymethylation, which is structurally similar to methylation but functionally different, is a biologically critical mechanism that is also known to regulate gene expression. Ten Eleven Translocation (TET) proteins mediate hydroxymethylation. However, the role of DNA hydroxymethylation or TETs in the endometrium remains relatively unexplored. Currently, the “gold standard” technique used to study methylation patterns is bisulfite genomic sequencing. This technique also detects hydroxymethylation but fails to distinguish between the two, thereby limiting our understanding of these two processes. The presence of TETs in the male and female reproductive tract and its contribution to endometrial cancer makes it an important factor to study in endometriosis. This review summarizes the role of DNA methylation in aberrant steroid hormone signaling and hypothesizes that hydroxymethylation could be a factor influencing hormonal instability seen in endometriosis.     

Disulfiram and Its Copper Chelate Attenuate Cisplatin-Induced Acute Nephrotoxicity in Rats Via Reduction of Oxidative Stress and Inflammation

Biological Trace Element Research - The use of cisplatin (CP) in chemotherapy of resistant cancers is limited due to its dose-dependent nephrotoxicity. Disulfiram (DSF), the aversion therapy for...