Crossing the Border: From Keto- to Imine Reduction in Short-Chain Dehydrogenases/Reductases

The family of NAD(P)H-dependent short-chain dehydrogenases/reductases (SDRs) comprises numerous biocatalysts capable of C=O or C=C reduction. The highly homologous noroxomaritidine reductase (NR) from Narcissus sp. aff. pseudonarcissus and Zt_SDR from Zephyranthes treatiae, however, are SDRs with an extended imine substrate scope. Comparison with a similar SDR from Asparagus officinalis (Ao_SDR) exhibiting keto-reducing activity, yet negligible imine-reducing capability, and mining the Short-Chain Dehydrogenase/Reductase Engineering Database indicated that NR and Zt_SDR possess a unique active-site composition among SDRs. Adapting the active site of Ao_SDR accordingly improved its imine-reducing capability. By applying the same strategy, an unrelated SDR from Methylobacterium sp. 77 (M77_SDR) with distinct keto-reducing activity was engineered into a promiscuous enzyme with imine-reducing activity, thereby confirming that the ability to reduce imines can be rationally introduced into members of the “classical” SDR enzyme family. Thus, members of the SDR family could be a promising starting point for protein approaches to generate new imine-reducing enzymes.     

Using an integrated process of data and modeling in HRA

The paper describes an approach taken to estimate the probabilities of failure associated with various railroad tasks to prevent accidents (principally collisions and derailments). These probabilities were estimated using an expert elicitation process that used partially relevant data available from a variety of databases and that were filtered and scaled to make them more directly relevant to the analyses being performed. Extensive qualitative studies were performed prior to the elicitation process to identify relevant contexts under which the tasks can be performed.     

Wavelength-tunable 40-GHz picosecond harmonically mode-locked fiber laser source

We demonstrate a novel 40-GHz mode-locked fiber laser that utilizes a single active device to provide both gain and mode-locking. The laser produces pulses as short as 2.2 ps, is tunable over a 27-nm band centered at 1553 nm, and exhibits long-term stability without cavity-length feedback control. The pulse train at 1556 nm was used in a 40-Gb/s transmission experiment over 45 km with a low 0.4-dB power penalty.     

Chicken acidic leucine-rich EGF-like domain containing brain protein (CALEB), a neural member of the EGF family of differentiation factors, is implicated in neurite formation

Chicken acidic leucine-rich EGF-like domain containing brain protein (CALEB) was identified by combining binding assays with immunological screens in the chicken nervous system as a novel member of the EGF family of differentiation factors. cDNA cloning indicates that CALEB is a multidomain protein that consists of an NH2-terminal glycosylation region, a leucine-proline-rich segment, an acidic box, a single EGF-like domain, a transmembrane, and a short cytoplasmic stretch. In the developing nervous system, CALEB is associated with glial and neuronal surfaces. CALEB is composed of a 140/130-kD doublet, an 80-kD band, and a chondroitinsulfate-containing 200-kD component. The latter two components are expressed in the embryonic nervous system and are downregulated in the adult nervous system. CALEB binds to the extracellular matrix glycoproteins tenascin-C and -R. In vitro antibody perturbation experiments reveal a participation of CALEB in neurite formation in a permissive environment.     

Transthyretin amyloidosis: a new mutation associated with dementia

NaCl-tolerant lactic acid bacteria (LAB) strains LC-10 (Lactobacillus casei) and LP-15 (Lact. plantarum) and NaCl were used as additives to sorghun (Sorghum bicolor). Numbers of LAB were significantly (P < 0.05) higher in all the additive-treated silages than in the control silage at an early stage of ensiling. During the fermentation process, addition of NaCl or LAB effectively inhibited the growth of aerobic bacteria and clostridia, but not yeasts. All the additive-treated silages had significantly (P < 0.05) lower pH, ammonia nitrogen content, dry matter loss and gas production but significantly (P < 0.05) higher lactic acid content and residual water soluble carbohydrates compared with the control silage. The improvement in silage quality was in the order: LAB > NaCl > control. Yeast counts were high in all additive-based silages and they increased during the exposure of the silages to air. As a result, these silages suffered aerobic deterioration, whereas the control silage was stable. The results confirmed that the NaCl or LAB improved fermentation quality but did not prevent aerobic deterioration of the silage.     

Visual results after submacular surgery for neovascularization in age-related macular degeneration

Orthopedists being more engaged in surgery are often asked about problems concerning exercise science. This article will summarize the existing basic knowledge, very close to practice, and put down the leading steps of how to build up a (therapeutical) training programme. A lab block is recommended acting on a preventive as well as on a sportive medical base.     

Tyrosine-dependent basolateral sorting signals are distinct from tyrosine-dependent internalization signals

Converting cysteine 543 to tyrosine in the influenza virus hemagglutinin (HA) introduces both a basolateral sorting signal and an internalization signal into the HA cytoplasmic domain. Another HA mutant, HA+8, contains eight additional amino acids at the end of the cytoplasmic domain that include a powerful internalization signal. HA+8 was also sorted efficiently to the basolateral surface of Madin-Darby canine kidney cells. The simplest explanation for the observation that multiple sorting phenotypes depend upon the same small amino acid sequence is that certain tyrosine-based internalization signals might also function as basolateral sorting signals. To test this hypothesis, second-site mutations were introduced into HA C543Y or HA+8 to determine if the internalization and basolateral sorting functions can be separated. For HA C543Y, the same sequence positions were important for both basolateral sorting and internalization, but the two functions responded differently to individual amino acid replacements, indicating that they were distinct. For HA+8, the basolateral sorting signal required the same tyrosine as the internalization signal, but did not share any other characteristics. Thus, even when basolateral sorting signals that depend on tyrosine overlap or are co-linear with internalizations signals, the two sorting processes are sensitive to different characteristics of the sequence.     

High-temperature sensible-heat storage options

Alkali-metal carbonate salts meet the requirements for high-temperature solar central receiver systems, but because of their corrosiveness they present special problems in the design of storage tanks. In order to reduce corrosion and temperature sufficiently to retain strength in the storage containing wall, internal and thermal insulation is required. We present design options and operation criteria for sensible-heat, molten-salt storage with internal insulation.     

Small thermophotovoltaic prototype systems

In an earlier paper we reported on a small grid-connected thermophotovoltaic (TPV) system consisting of an ytterbia mantle emitter and silicon solar cells with 16% efficiency (under solar irradiance in standard test conditions, STCs). The emitter was heated up using a butane burner with a rated thermal power of 1.35 kW (referring to the lower heating value). This system produced an electrical output of 15 W, which corresponds to a thermal to electric (direct current) conversion efficiency of 1.1%. In the interim, further progress has been made, and significantly higher efficiencies have been achieved. The most important development steps are: (1) the infrared radiation-absorbing water filter between emitter and silicon cells (to protect the cells against overheating and against contact with flue gasses) has been replaced by a suitable glass tube. By doing this, it has been possible to prevent losses of convertible radiation in water. (2) Cell cooling has been significantly improved, in order to reduce cell temperature, and therefore increase conversion efficiency. (3) The shape of the emitter has been changed from spherical to a quasi-cylindrical geometry, in order to obtain a more homogeneous irradiation of the cells. (4) The metallic burner tube, on which the ytterbia emitter was fixed in the initial prototypes, has been replaced by a heat-resistant metallic rod, carrying ceramic discs as emitter holders. This has prevented the oxidation and clogging of the perforated burner tube. (5) Larger reflectors have been used to reduce losses in useful infrared radiation. (6) Smaller cells have been used, to reduce electrical series resistance losses. Applying all these improvements to the basic 1.35 kW prototype, we attained a system efficiency of 1.5%. By using preheated air for combustion (at approximately 370 °C), 1.8% was achieved. In a subsequent step, a photocell generator was constructed, consisting of high-efficiency silicon cells (21% STC efficiency). In this generator, the spaces between the cells were minimized, in order to achieve as high an active cell area as possible, while simultaneously reducing radiation losses. This new system has produced an electrical output of 48 W, corresponding to a system efficiency of 2.4%. This is the highest-ever-reported value in a silicon-cell-based TPV system using ytterbia mantle emitters. An efficiency of 2.8% was achieved by using preheated air (at approximately 500 °C). An electronic control unit (fabricated of components with low power consumption, and including a battery store) was developed, in order to make the TPV system self-powered. This unit controls the magnetic gas supply valve between gas supply cylinder and burner as well as the high-voltage ignition electrodes. Both the control unit’s own power consumption and the battery-charging power are supplied directly by the TPV generator. A small commercial inverter is used to transfer excess power to the 230 V grid.     

High Throughput Primer Walking of cDNA Clones

Primer walking of cloned DNA is a standard research tool. It has been used in the past to determine the sequence of individual clones of interest. With the expansion of DNA sequencing capacity the need to be able to walk larger numbers of clones has become necessary. Our laboratory is a mid-sized genomics facility. In conjunction with the Advanced Biomedical Computing Center (ABCC) we have developed methods for automating the primer selection, DNA sequencing, contig assembly and sequence analysis for clones arrayed in microtiter format. This approach has allowed us to walk 475 clones (five microtiter plates) selected from a cDNA library.