Extending the carbon chain: hydrocarbon formation catalyzed by vanadium/molybdenum nitrogenases

In a small-scale reaction, vanadium-dependent nitrogenase has previously been shown to catalyze reductive catenation of carbon monoxide (CO) to ethylene, ethane, propylene, and propane. Here, we report the identification of additional hydrocarbon products [α-butylene, n-butane, and methane (CH(4))] in a scaled-up reaction featuring 20 milligrams of vanadium-iron protein, the catalytic component of vanadium nitrogenase. Additionally, we show that the more common molybdenum-dependent nitrogenase can generate the same hydrocarbons from CO, although CH(4) was not detected. The identification of CO as a substrate for both molybdenum- and vanadium-nitrogenases strengthens the hypothesis that CO reduction is an evolutionary relic of the function of the nitrogenase family. Moreover, the comparison between the CO-reducing capacities of the two nitrogenases suggests that the identity of heterometal at the active cofactor site affects the efficiency and product distribution of this reaction.     

Requirement of AMPA receptor GluR2 phosphorylation for cerebellar long-term depression

Cerebellar long-term depression (LTD) is a model of synaptic memory that requires protein kinase C (PKC) activation and is expressed as a reduction in the number of postsynaptic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors. LTD was absent in cultured cerebellar Purkinje cells from mutant mice lacking the AMPA receptor GluR2 subunit and could be rescued by transient transfection with the wild-type GluR2 subunit. Transfection with a point mutant that eliminated PKC phosphorylation of Ser880 in the carboxy-terminal PDZ ligand of GluR2 failed to restore LTD. In contrast, transfection with a point mutant that mimicked phosphorylation at Ser880 occluded subsequent LTD. Thus, PKC phosphorylation of GluR2 Ser880 is a critical event in the induction of cerebellar LTD.     

Distinguishing inchworm and hand-over-hand processive kinesin movement by neck rotation measurements

The motor enzyme kinesin makes hundreds of unidirectional 8-nanometer steps without detaching from or freely sliding along the microtubule on which it moves. We investigated the kinesin stepping mechanism by immobilizing a Drosophila kinesin derivative through the carboxyl-terminal end of the neck coiled-coil domain and measuring orientations of microtubules moved by single enzyme molecules at submicromolar adenosine triphosphate concentrations. The kinesin-mediated microtubule-surface linkage was sufficiently torsionally stiff (>/=2.0 +/- 0.9 x 10(-20) Newton meters per radian2) that stepping by the hypothesized symmetric hand-over-hand mechanism would produce 180 degree rotations of the microtubule relative to the immobilized kinesin neck. In fact, there were no rotations, a finding that is inconsistent with symmetric hand-over-hand movement. An alternative "inchworm" mechanism is consistent with our experimental results.     

Molecular sorting in the secretory pathway

Proteins can be secreted from animal cells by either a constitutive or a regulated pathway; those destined for regulated secretion are actively sorted into dense-core secretory granules. Although sorting is generally assumed to be accomplished by specific carriers, the nature of these carriers remains elusive. In this study, peptide hormones were used as affinity ligands to purify a set of 25-kilodalton proteins from canine pancreatic tissue. Their ligand specificities and patterns of expression have the characteristics of sorting carriers.     

Three-dimensional porous collagen/chitosan complex sponge for tissue engineering

A three-dimensional, porous collagen/chitosan complex sponge was prepared to closely simulate basic extracellular matrix (ECM) constitutes, collagen and glycosaminoglycan. The complex sponge was prepared by a lyophilization method and had the regular network with highly porous structure, suitable for cell adhesion and growth. The pores were well interconnected, and their distribution was fairly homogeneous. The complex sponge was crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to increase its biological stability and enhance its mechanical properties. The crosslinking medium had a great effect on the inner structure of the sponge. The homogeneous, porous structure of the sponge was remarkably collapsed in an aqueous crosslinking medium. However, the morphology of the sponge remained almost intact in a water/ethanol mixture crosslinking milieu. Mechanical properties of the collagen/chitosan sponge were significantly enhanced by EDC-mediated crosslinking. The potential of the sponge as a scaffold for tissue engineering was investigated using a Chinese hamster ovary cell (CHO-K1) line.     

Neck formation in drawing processes of fibers

To better understand the formation of necking in drawing processes of fibers, strain distributions during drawing processes have been analyzed. For simplicity, one-dimensional incompressible steady flow at a constant temperature was assumed and quasi-static model was used. To describe mechanical properties of solid polymers, non-linear visco-plastic material properties were assumed using the power law type hardening and rate-sensitive equation. The effects of various parameters on the neck formation were mathematically analyzed. As material property parameters, strain-hardening parameter, visco-elastic coefficient and strain-rate sensitivity were considered and, for process parameters, the drawing ratio and the process length were considered. It was found that rate-insensitive materials do not reach a steady flow state and the rate-sensitivity plays a key role to have a steady flow. Also, the neck formation is mainly affected by material properties, especially for the quasi-static model. If the process length changes, the strain distribution was found to be proportionally re-distributed along the process line by the factor of the total length change.     

Compositional and digestibility changes in sprouted barley and canola seeds

Barley and canola seeds were sprouted over a 5 day period, in laboratory conditions under room temperature (22°C) and room lighting. Following initial hydration, seeds were kept moist by wetting the germination trays at 9 a.m., 1 p.m. and 6 p.m. daily. A parallel germination experiment using 200 g quantities of seeds in petri dishes was conducted. Starting from the second day of germination, and every day, dishes of germinating seeds were removed, oven-dried, weighed and milled for proximate and chemical analysis. Seeds from the main germination experiment were fed in a digestibility trial to Wistar rats. Results indicated that sprouting was associated with depletion of many nutrients in both barley and canola, the major losses being in respect of dry matter, gross energy and triglycerides. In barley (but not in canola) sprouting was associated with significant increases in crude fiber and diglyceride content. In canola, there were significant losses in lipid content and increases in phytosterol and phospholipid content. Digestibility data showed an enhancement in digestibility of nutrients in barley but not in canola, implying that sprouting improved nutritional quality of barley but not canola.     

On-line monitoring of cellulase treatment by differential refractometer

Cellulase treatment of cellulose fibers needs to be monitored to give proper weight loss without significant strength loss. On-line monitoring of cellulase treatment is presented which can monitor the weight loss of cotton fabrics by measuring differential refractive index in real time. On-line monitoring was tried under the condition where the cellulase treatment of scoured 100% cotton knits would give weight loss of about 6.6% during one hour cellulase treatment. For comanalyses processing solutions with treatment time of 5, 10, 20, 30, 45 and 60 minutes were taken out, and subjected to sugar analyses using total organic carbon (TOC), and dinitrosalicylic acid (DNS) method as well as HPLC analyses. HPLC analyses showed that cellobiose and glucose were the major components resulting from cellulase treatment of cotton fabric. TOC and differential refractive index measurement proved to be linear to total sugar concentration while DNS method result was dependent on the type of sugar. Various sugar concentrations of the processing solutions measured by these methods were compared with HPLC measurements and correlated with the actual weight loss of the cotton fabric processed. The on-line monitoring device using differential refractometer gave a real time signal which was independent of the ratio of cellobiose and glucose but dependent on the total sugar concentration. The device was also non-destructive.     

Nonsteady plane-strain ideal forming without elastic dead-zone

Ever since the ideal forming theory has been developed for process design purposes, application has been limited to sheet forming and, for bulk forming, to two-dimensional steady flow. Here, application for the non-steady case was made under the plane-strain condition. In the ideal flow, material elements deform following the minimum plastic work path (or mostly proportional true strain path) so that the ideal plane-strain flow can be effectively described using the two-dimensional orthogonal convective coordinate system. Besides kinematics, schemes to optimize preform shapes for a prescribed final part shape and also to define the evolution of shapes and frictionless boundary tractions were developed. Discussions include numerical calculations made for a real automotive part under forging.     

Leukotriene C4 release and gene expressions of IL-8 and MCP-1 in porcine alveolar epithelial type II cells

Leukotrienes (LT) and chemokines are important chemotactic compounds in regulating the recruitment and activation of immune cells during pulmonary inflammatory reactions. Results showed that LTC4 release by porcine alveolar epithelial type II cells (AEC IIs) is significantly enhanced by either LTB4 or LPS stimulation. The basal level of IL-8 gene expression in AEC IIs was only 1/3 of that observed in alveolar macrophages (AMs) while AEC IIs expressed a higher basal level of monocyte chemotactic peptide-1 (MCP-1) and also in response to LPS stimulation than do AMs. The increasing basal and LT-induced MCP-1 gene expressions after 8h of incubation were observed in AEC IIs but decreased in AMs. These findings suggest that AEC IIs play an important role in initial inflammatory reactions of the lung by releasing LTC4, and that they also modulate later inflammatory reactions, evidenced by consistent elevation of MCP-1 gene expression after and during exogenous challenge in pigs.