2.4 kW 1045 nm narrow-spectral-width monolithic single-mode CW fiber laser by using an FBG-based MOPA configuration.

A 24 kW narrow-spectral-width near-diffraction-limited monolithic fiber laser system at ${\sim}{1045.2}\;{\rm{nm}}$ in a fiber Bragg grating (FBG)-based master oscillator power amplifier (MOPA) configuration is demonstrated in this paper. The near-diffraction-limited beam quality (${{\rm{M}}^2}\sim{1.2}$) and a spectral width of 0.35 nm (${\sim}{{96}}\;{\rm{GHz}}$) are achieved. The stimulated Raman scattering (SRS) is theoretically and experimentally investigated. The SRS has been suppressed by carefully optimizing the length of the Yb-doped fiber and the pumping scheme, and a signal-to-noise ratio of ${\sim}{{33}}\;{\rm{dB}}$ between the laser signal and the Raman Stokes component is achieved. The stimulate Brillouin scattering and the transverse mode instability are not observed. To our best knowledge, this is the highest-output power for ${{104}} \times {\rm{nm}}$ single-mode fiber laser with ${\sim}{{96}}\;{\rm{GHz}}$ spectral width by using an FBG-based MOPA configuration.

Density Functional Theory-Assisted Electrochemical Assay Manipulated by a Donor-Acceptor Structure toward Pharmaceutical Diagnostic.

Oxidative stress is a state of stress injury, which leads to the pathogenesis of most neurodegenerative diseases. Moreover, this is also one of the main reasons for the loss of dopaminergic neurons and the abnormal content of dopamine (DA). In the past decades, a number of studies have found that acetaminophen (AP) is metabolized and distributed in the brain when it is used as a neuroprotective compound. In this context, we proposed an electrochemical sensor based on 9-(4-(10-phenylanthracen-9-yl)phenyl)-9H-carbazole with the goal of diagnosing these two drugs in the body. Carbazole groups can easily be formed into large π-conjugated systems by electropolymerization. The introduction of anthracene exactly combined the carbazole group to establish an efficient electron donor-acceptor pattern, which enhanced π-π interaction with the electrode surface and charge transporting ability. The diagnostic platform showed good sensing activity toward the oxidation of DA and AP. The detection range for DA and AP is from 0.2 to 300 µM and from 0.2 to 400 µM, respectively. The simultaneous detection range is from 0.5 to 250 µM, which is wider than most reports. After a series of electrochemical assessments were determined, the sensor was finally developed to the analysis of pharmaceutical and human serum, displaying a meaningful potential in clinical evaluation.

An electrochemical thrombin aptasensor based on the use of graphite-like C3N4 modified with silver nanoparticles.

An electrochemical aptasensor for thrombin is introduced that makes use of a nanohybrid composed of silver nanoparticles and graphite-like carbon nitride (Ag-g-C3N4). The material has a large surface and good biocompatibility. AgNPs are modified directly on the surface of g-C3N4 via chemical reduction. A glass carbon electrode (GCE) modified with Ag-g-C3N4 can immobilize a large number of amino-terminated thrombin binding aptamers (NH2-TBA) through strong Ag-N bonds. The electrochemical impedance signal of the aptasensor increases in the presence of thrombin. Under the optimal conditions and by using [Fe(CN)6]3-/4- as an electrochemical probe, the aptasensor shows a wide linear range of 100 fM - 20 nM with a lower detection limit of 38 fM. The method was applied to the determination of thrombin in spiked human plasma and the recoveries fluctuated from 97.2% to 103%. Graphical abstractSchematic representation of an electrochemical aptasensor using graphite-like carbon nitride (C3N4) modified with silver nanoparticles as electrode substrate for thrombin (TB) detection.

The synergistic effect of Au-COF nanosheets and artificial peroxidase Au@ZIF-8(NiPd) rhombic dodecahedra for signal amplification for biomarker detection.

Here, a new type of signal amplification strategy is proposed employing Au nanoparticle (AuNP)-functionalized covalent organic framework (Au-COF) nanosheets and AuNP functionalized ZIF-8(NiPd) (Au@ZIF-8(NiPd)) rhombic dodecahedra nanocomposites for sandwich electrochemical sensor construction. The peroxidase mimics Au@ZIF-8(NiPd) took the place of natural enzymes in enzyme-assisted amplification strategies, both acting as catalysts for H2O2 reduction for signal amplification, and serving as ideal nanocarriers for signal probe anchoring. The cancer biomarker thrombin (TB) was selected as the target. Thrombin binding aptamers (TBA 2) were fixed on Au@ZIF-8(NiPd), and the obtained TBA 2-Au@ZIF-8(NiPd) bioconjugates were employed as tracer labels, and TB was sandwiched between the tracer labels and capture probe TBA 1 which were immobilized on the Au-COF nanosheet modified electrode. Au-COFs with a high specific area, super electroconductivity, and uniformly distributed AuNPs were utilized as the electrode substrate to fix TBA 1. Exploiting the sandwich method, the proposed TB aptasensor exhibited a wide linear range of 0.1 pM to 20 nM with a low detection limit of 15 fM (S/N = 3). The ingenious sensing strategy enriched the application diversity of the artificial enzyme and showed promise in research and development of point-of-care diagnostics.

Charge Transfer Platform and Catalytic Amplification of Phenanthroimidazole Derivative: A New Strategy for DNA Bases Recognition.

Research about DNA composition has been concentrated on DNA damage in the past few decades. However, it still remains a great challenge to construct a rapid, facile, and accurate approach for simultaneously monitoring four DNA bases, guanine (G), adenine (A), thymine (T), and cytosine (C). Herein, a novel electrochemical sensor based on phenanthroimidazole derivative, 2-(4-bromophenyl)-1-phenyl-1H-phenanthro[9,10-d]-imidazole (PPI), is successfully fabricated by a simple electrochemical method. The bromophenyl group in PI could expand their aromatic plane, induce the π-conjugated extension, and enhance the charge transfer and π-π interaction. The phenyl group at N1 position could regulate the intermolecular interaction, which could promote the possibility of intermolecular connection. The PPI polymer (poly(PPI)) with π-electron enriched conjugation architecture has been applied in simultaneous determination of G, A, T, and C in neutral solution by square wave voltammetry (SWV) method with well-separated peak potentials at 0.714, 1.004, 1.177, and 1.353 V, respectively. The sensor functionalized with poly(PPI) exhibits wide linear response for G, A, T, and C in the concentration ranges of 3-300, 1-300, 30-800, and 20-750 µM, respectively. With favorable selectivity, stability, and reproducibility, the sensor is successfully utilized to monitor four DNA bases in real samples, displaying a promising prospect for electrochemical sensing devices.

AuPt/MOF-Graphene: A Synergistic Catalyst with Surprisingly High Peroxidase-Like Activity and Its Application for H2O2 Detection.

Here, we report ZIF-8-reduced graphene oxide (ZIF-8-rGO)-supported bimetallic AuPt nanoparticles (AuPtNPs) as a novel peroxidase mimic for high-sensitivity detection of H2O2 in neutral solution. ZIF-8-graphene oxide (ZIF-8-GO) is first synthesized via a simple wet-chemistry process and subsequently immobilized with AuPtNPs via a reduction method. The resultant AuPt/ZIF-8-rGO shows enhanced peroxidase-like catalytic activity and it is applied for the electrochemical detection of H2O2 in a wide concentration range, from 100 nM to 18 mM, with a very low detection limit of 19 nM (S/N = 3). This good electroanalytical performance of AuPt/ZIF-8-rGO is owing to the ultrasmall size and high dispersion of the AuPtNPs, the strong metal-support interaction between the AuPtNPs and ZIF-8-rGO bisupport, and the sandwich-like structure comprising porous ZIF-8 and loosely packed rGO nanosheets. The AuPt/ZIF-8-rGO is employed for the practical detection of H2O2 in human serum samples with desirable properties. Therefore, the novel AuPt/ZIF-8-rGO is a promising nanozyme for various biotechnological and environmental applications.

New Strategy for Ultrasensitive Aptasensor Fabrication: D-A-D Constitution as a Charge Transfer Platform and Recognition Element.

Over the past decade, various sensing systems based on aptamers have attracted a great deal of studies directed at designing highly selective biosensors. In this paper, we described a new-style electrochemical aptamer sensor (aptasensor) via a donor-acceptor link substrate, which was characterized by electrochemical methods and other helpful characterization instruments. Molecules with D-A-D configuration always undergo an intrinsic signal amplification due to the elongation of the π-electron conjugation. Triphenylamine, a peripheral electron donor, has excellent hole-transport property and is able to assemble on the surface of glassy carbon electrode by π-π stacking interaction. To further improve the performance of the adenosine triphosphate (ATP) sensor, we chose diphenylfumaronitrile-containing electron-withdrawing group as the central core to promote charge transfer, which can also efficiently combine with aptamers by multihydrogen bond function. Surprisingly, the sensing platform showed a wide liner range from 0.1 pM to 100 nM, with a detection limit of 0.018 pM. We examined the ATP in human serum sample, indicating that the novel aptasensor based on D-A-D conjugated polymer holds great possibility for practical detection of ATP. Moreover, it is foreseeable that the conjugated polymers of the D-A structure will have promising application in the preparation of biosensors.

A bioinspired antifouling zwitterionic interface based on reduced graphene oxide carbon nanofibers: electrochemical aptasensing of adenosine triphosphate.

An antifouling electrochemical aptasensor for ATP is described that has a zwitterionic self-assembled sensing interface on a glassy carbon electrode modified with a reduced graphene oxide carbon nanofiber (GO-CNF). The GO-CNF was first modified by self-polymerization of dopamine which provided a platform for simultaneously self-assembly of the ATP aptamer and cysteine. By using hexacyanoferrate as the electrochemical probe, in the presence of ATP, the aptamer strands fold around ATP molecules, thus leading to the variation of the electrochemical signal. The aptasensor has a linear response in the 0.1 pM to 5 nM ATP concentration range, and a 13 fM lower detection limit. The electrode is strongly resistant to nonspecific adsorption and biofouling. This enabled the detection of ATP even in spiked human plasma. Graphical abstract An antifouling electrochemical aptasensor employing reduced graphene oxide carbon nanofiber as conductive substrate and zwitterionic cysteine as antifouling material for adenosine triphosphate detection.

Facile synthesis of 3D N-doped porous carbon nanosheets as highly active electrocatalysts toward the reduction of hydrogen peroxide.

Constructing three-dimensional (3D) conductive frameworks with high specific surface areas and porous structures is indispensable for their applications as electrocatalysts. In this work, we illustrate for the first time that 3D N-doped porous carbon nanosheets (3D-NS), which are synthesized via a facile one-pot pyrolysis reaction using glucose and melamine as raw materials, can serve as high performance and green electrocatalysts for the reduction of H2O2. Moreover, a series of 3D-NS samples with a controllable content of nitrogen were obtained by adjusting the calcination temperature. From our research, the 3D-NS obtained at 900 °C possessed high specific surface areas, porous structures, proper dosages of N atoms, suitable degrees of graphitization and defects. Furthermore, we also illustrate their application in H2O2 electrochemical sensing in physiological environments. Under optimum conditions, the 3D-NS-based sensor displays a wide linear scope in the range of 0.5-14 000 µM and a low detection limit of 0.18 µM (S/N = 3). Therefore, with desirable selectivity, stability and anti-interference performance, the proposed sensor can be feasibly applied to detect H2O2 in human serum samples.

Signal amplification strategy for biomarkers: Structural origins of epitaxial-growth twinned nanocrystals and D-π-A type polymers.

The combination of nanoparticles and biomarkers yields functional nanostructured biointerface, which is playing a notable role in biotechnology development. Due to the 5-fold twined structure in the Au-Pt star-shaped decahedra not only allowed it to act as efficient scaffold for immobilization of antibody, but it also exhibits superior electrocatalytic activity toward H2O2 reduction, the nanocrystal as the efficient signal transduction label is first employed to construct an electrochemical immunosensor. Donor-π-Acceptor (D-π-A) linking fashion generates a dipolar push-pull system and assures superior intramolecular charge transfer. It is considered as a suitable π-conjugated backbone for conducting polymer on biointerface application. Under a D-π-A architecture which imidazole as the π-bridge and amino phenyl/phenyl groups as peripheral electron-donating/withdrawing functional groups, 4-(2,4,5-triphenyl-1H-imidazol-1-yl) aniline (TPIDA) is designed and synthesized for good biocompatibility and high conductivity. In this proposal, we attempt to integrate the above-mentioned two features from nanobiotechnology and organic bioelectronics. Then, a novel nonenzymatic sandwich-type immunosensor is performed by Au-Pt core-shell with surface-engineered twinning as a label and π-conjugated D-π-A polymers as the signal amplification platform. Human IgG (HIgG) as the model target protein can be detected with a wide linear range from 0.1 pg mL-1 to 100 ng mL-1. The detection limit is down to 0.06 pg mL-1 (S/N = 3). Moreover, as a practical application, the prepared biosensor is used to monitor HIgG level in human serum with desirable results obtained.

Analysis of mode-hop free tuning of folded cavity grating feedback lasers.

We analytically study an external cavity laser structure including a folded cavity. A steering mirror is utilized in the folded cavity to deflect the intracavity laser beam. A mode-hop free tuning range of ∼400 GHz can be achieved by control of the steering mirror, and a fast tuning rate is expected because of the small mass of the steering mirror. This technique has potential for applications in spectroscopy for turbulent media, especially in the mid-infrared region.

Nurse eggs form through an active process of apoptosis in the spionid Polydora cornuta (Annelida).

The production of nurse eggs is fundamental to poecilogony in some species of spionid annelids. In species such as Polydora cornuta, nurse-egg production varies among females and ingestion of nurse eggs varies among young, resulting in a form of poecilogony with divergent phenotypes for females (e.g., fecundity and per-offspring investment) as well as for larvae (e.g., trophic mode, size, and stage at hatching). We tested the hypothesis that nurse eggs of P. cornuta form through an active developmental process and specifically, through apoptosis. Results of a TUNEL assay indicate nuclear fragmentation occurs in a process that is characteristic of apoptosis. Cellular indicators of apoptosis in nurse eggs include activation of caspase-3, a positive Annexin V reaction indicating exposure of phosphatidylserine on the outer cell membrane, and invagination of the membrane to form yolk vesicles. These results indicate that formation of nurse eggs in this population of P. cornuta occurs through an active, adaptive process. Furthermore, while apoptosis also occurs in some cells of P. cornuta embryos, it was not detected until later in development. This suggests that nurse eggs originate through heterochrony in a developmental process (apoptosis) that is common to all young of P. cornuta.

Elastic properties of the cell wall of Aspergillus nidulans studied with atomic force microscopy.

Currently, little is known about the mechanical properties of filamentous fungal hyphae. To study this topic, atomic force microscopy (AFM) was used to measure cell wall mechanical properties of the model fungus Aspergillus nidulans. Wild type and a mutant strain (deltacsmA), lacking one of the chitin synthase genes, were grown in shake flasks. Hyphae were immobilized on polylysine-coated coverslips and AFM force--displacement curves were collected. When grown in complete medium, wild-type hyphae had a cell wall spring constant of 0.29 +/- 0.02 N/m. When wild-type and mutant hyphae were grown in the same medium with added KCl (0.6 M), hyphae were significantly less rigid with spring constants of 0.17 +/- 0.01 and 0.18 +/- 0.02 N/m, respectively. Electron microscopy was used to measure the cell wall thickness and hyphal radius. By use of finite element analysis (FEMLAB v 3.0, Burlington, MA) to simulate AFM indentation, the elastic modulus of wild-type hyphae grown in complete medium was determined to be 110 +/- 10 MPa. This decreased to 64 +/- 4 MPa for hyphae grown in 0.6 M KCl, implying growth medium osmotic conditions have significant effects on cell wall elasticity. Mutant hyphae grown in KCl-supplemented medium were found to have an elastic modulus of 67 +/- 6 MPa. These values are comparable with other microbial systems (e.g., yeast and bacteria). It was also found that under these growth conditions axial variation in elastic modulus along fungal hyphae was small. To determine the relationship between composition and mechanical properties, cell wall composition was measured by anion-exchange liquid chromatography and pulsed electrochemical detection. Results show similar composition between wild-type and mutant strains. Together, these data imply differences in mechanical properties may be dependent on varying molecular structure of hyphal cell walls as opposed to wall composition.

Purification and characterization of mannitol dehydrogenase and identification of the corresponding cDNA from the head blight fungus, Gibberella zeae (Fusarium graminearum).

The mannitol-2-dehydrogenase (MtDH) from Gibberella zeae was purified and the corresponding cDNA identified. Purification of MtDH was accomplished using a combination of ammonium sulfate fractionation, anion exchange and dye-ligand chromatography. Final purification was achieved following electroelution from a native gel. Molecular mass determination based on SDS-PAGE indicated that the denatured protein was 29 kDa. Native protein mass was determined to be 110 kDa using gel permeation chromatography, indicating a tetrameric form. The pH optima for mannitol oxidation and fructose reductase activities were 9.0, and 7.0, respectively. Activity with sorbitol as the substrate was 21% of activity with mannitol. Kinetic parameters were determined by direct-linear plots of enzyme activity vs. substrate concentrations. Fructose concentrations above 600 mM and NADPH concentrations above 0.3 mM caused substrate inhibition. Comparisons of predicted amino acid sequences of several fungal MtDHs indicated high conservation within the phyla. A possible role for MtDH in generation of turgor pressure for forcible ascospore discharge is discussed.

Integrated bioprocessing in Saccharomyces cerevisiae using green fluorescent protein as a fusion partner.

In this study, we examine the use of green fluorescent protein (GFP) for monitoring a hexokinase (HXK)-GFP fusion protein in Saccharomyces cerevisiae for various events including expression, degradation, purification, and localization. The fusion, HXK-EK-GFP-6 x His, was constructed where the histidine tag (6 x His) would allow for convenient affinity purification, and the enterokinase (EK) cleavage site would be used for separation of HXK from GFP after affinity purification. Our results showed that both HXK and GFP remained active in the fusion and, more importantly, that there was a linear correlation between HXK activity and GFP fluorescence. Enterokinase cleavage studies revealed that both GFP fluorescence intensity and HXK activity remained unchanged after separation of the fusion proteins, which indicated that fusion of GFP did not cause structural alteration of HXK and thus did not affect the enzymatic activity of HXK. We also found that degradation of the fusion protein occurred, and that degradation was limited to HXK with GFP remaining intact in the fusion. Confocal microscopy studies showed that while GFP was distributed evenly in the yeast cytosol, HXK-GFP fusion followed the correct localization of HXK, which resulted in a di-localization of both cytosol and the nucleus. GFP proved to be a useful fusion partner that may lead to the possibility of integrating the bioprocesses by quantitatively following the entire process visually.

Impediments to secretion of green fluorescent protein and its fusion from Saccharomyces cerevisiae.

While it has been demonstrated that GFP-tagged proteins were transported to their correct cellular compartments in most cells, attempts to secrete GFP/GFP-fusion through the default secretory pathway have not been as successful. In an attempt to induce secretion of GFP and Hexokinase (HXK)-GFP fusion in Saccharomycescerevisiae, we have cloned constructs that employed four different yeast secretion signal sequences, i.e., INU1, SUC2, PHO5, and MEL1. The expression is under the control of the galactose-inducible GAL1 promoter. Our results showed that all eight constructs entered the secretory pathway successfully, and the signal peptides were all cleaved off. However, none of the eight constructs were able to lead to secretion into the culture media or the periplasmic space. The expression levels of the eight constructs differ dramatically, depending on both the signal peptide and whether GFP was fused with HXK. Confocal microscopy studies revealed that the eight constructs also led to very different localization patterns. Among them, two constructs targeted GFP to the vacuole partially or exclusively, whereas others were mostly retained in the ER/Golgi compartments. Our efforts, together with those of others, seem to suggest that the signal peptide itself is not enough to lead to secretion of GFP from S. cerevisiae, although it has been successful in some other organisms. Nonetheless, the advantage of GFP's in vivo detection makes it a powerful tool for investigating protein localization events.

Physiological and environmental aspects of ascospore discharge in Gibberella zeae (anamorph Fusarium graminearum).

We investigated ascospore discharge in the perithecial fungus, Gibberella zeae. In a wind tunnel study that simulated constant rain and varying day and night lengths, the rate of ascospore release was approximately 8-30% greater under light than in complete darkness. Under constant light, ascospore discharge occurred at maximal rates at relative humidity levels greater than 92%. When perithecia were placed under conditions of high external osmolarity, ascospore discharge was significantly reduced. Ascospores were discharged from asci along with droplets of fluid, the epiplasm, from within the ascus. Analysis of discharged epiplasmic fluid by GC-MASS Spectrometry revealed that mannitol was the major simple sugar component of the fluid. Activity of mannitol dehydrogenase, which catalyzes the conversion of fructose to mannitol, was higher in protein extracts from mature perithecia than in extracts from vegetative tissue. Several inhibitors of K(+) and Ca(++) ion channels inhibited ascospore discharge, which suggested that ascospore discharge resulted from the buildup of turgor pressure generated by ion fluxes and mannitol accumulation.