The conventional short-circuiting technique under-short-circuits most epithelia

Summary The relationships among ion current, membrane potential difference, and resistance of an epithelium are studied. The short-circuit technique introduced by Ussing and Zerahn does not completely short circuit the epithelium if the series resistance parallel to the cell layer between the voltage electrodes is not properly compensated. The residual potential difference across the epithelial cell layer in the short-circuit state is proportional to both the measured short-circuit current and the resistance of the diffusion barriers not compensated. In the conventionally short-circuited small intestinal mucosa the villus and crypt areas are hypo-polarized to different degrees rather than simultaneously hyper- and hypo-polarized. Short-circuiting the whole tissue reduces but does not abolish the passive net ion movement across the tissue. Measurements of the electrical properties of the whole and denuded rat distal small intestine in HCO₃-Ringer solution containing 10mm glucose reveal that the measured short-circuit current has under-estimated approximately 33% of the true short-circuit current and that the passive net Na flux from serosa to mucosa and Cl flux from mucosa to serosa are not negligible in the short-circuit state.     

Release of arachidonate from diglyceride in human platelets requires the sequential action of a diglyceride lipase and a monoglyceride lipase

Release of arachidonate from 2-arachidonyl diglyceride by human platelet microsomes was investigated. Diglycerides labeled with ¹⁴C-stearate at sn-1 and with ³H-arachidonate at sn-2 were used as a substrate for microsomal diglyceride lipase. Diglyceride was deacylated first at sn-1 as evidenced by the accumulation of 2-arachidonyl monoglyceride but not of 1-stearoyl monoglyceride. Subsequent release of arachidonate from monoglyceride required the action of a monoglyceride lipase. Studies on substrate specificity indicated that diglyceride lipase utilized 2-arachidonyl diglyceride as the best substrate.     

Triphasic concentration effects of gentamicin on activity and misreading in protein synthesis.

Gentamicin is shown to exert a triphasic concentration effect on peptide synthesis in vitro with natural messengers. Low concentrations (up to 2 micron) caused slowing and a decrease in total synthesis, but little misreading (assayed with extracts lacking Glu-tRNA); the inhibition was greater with an initiating system (with phage RNA as messenger) than with pure chain elongation on purified endogenous polysomes of Escherichia coli. Moderate concentrations (up to 100 micron) slowed synthesis less, markedly increased its duration in the noninitiating system, and strongly stimulated misreading; at optimal concentrations total synthesis was even greater than normal. Moreover, with phage RNA these concentrations increased the synthesis of large polypeptides. We conclude that binding of gentamicin to its first site causes inhibition but little misreading; binding to additional site(s) partly reverses the inhibition by first-site binding and markedly stimulates misreading, and the misreading appears to favor "readthrough" of termination codons. In the third phase (greater than 100 micron) synthesis is slowed again but the pattern of misreading does not appear to be altered; this effect need not involve a specific further action on the ribosome.     

Hydrolase production by Aspergillus niger in solid-state cultivation

Summary A production of macerating enzymes which liquefy and hydrolyze the mandarin orange peel was studied in a solid state cultivation of Aspergillus niger on wheat bran substrate. Solid state cultivation in a 2 drum fermenter capable of interchangeable operation under dynamic or static conditions were carried out maintaining the moisture content of the substrate at 32, 39, 46, 56, 67, and 74%. Biomass grown on the solid substrate was estimated on the basis of a constant value of glucosamine content of A. niger, 50 mg glucosamine/g cell. A linear relationship between oxygen uptake rate and growth rate observed in all the experiments gave an oxygen growth yield, Y_X/O, of 28.5 g cell/mol O₂. The rate of macerating enzyme formation was also in proportion to the growth rate irrespective of the difference of the moisture content of the substrate.The enzyme accumulation on the solid substrate, the growth rate and oxygen uptake rate were maximum when the moisture content of the substrate was maintained at ca. 56% ascending from 32 to 56 and descending from 56 to 74.     

Primary structure of streptococcal proteinase. III. Isolation of cyanogen bromide peptides: complete covalent structure of the polypeptide chain.

The following sequence has been derived for streptococcal proteinase. (See article). The sequence permits the assignment of the single cysteine residue essential for catalytic action at position 47 from the NH2 terminus of the protein. The tryptophan residue at the binding site of the enzyme is at position 214. A histidine residue at position 195 has been assigned as the catalytically important entity in the molecule. Streptococcal proteinase and papain, an enzyme with similar properties, are compared with respect to structure and function.     

A visit to the zoos and aquariums in Northern Japan

For their trip to the National Parks in Hokkaido the authors also included in their schedule visits to the zoos and aquariums in Northern Japan. Their tight scheduled tour started at the Aquamarine Fukushima, which was after the large tsunami rebuilt and re-opened in July 2011. They continued with a visit to the Kamo Aquarium, which is famous for its unique and large jellyfish exhibition. On Hokkaido Island they visited the smaller zoos in Kushiro and Obihiro with a good collection of native Hokkaido animal species. Included was also the Asahiyama Zoo, which is at the moment one of the most popular zoos in Japan. It is well-known for its innovative enclosures for seals and penguins. The Sapporo Maruyama Zoo and the Chitose Salmon Aquarium were the last highlights of this tour. The themes and animal collections of these different zoos and aquariums are described. The general situation for the northern zoos and aquariums is quite different compared to the institutions in the southern parts of the country and entirely to the zoos in the large city areas like Tokyo, Osaka and Nagoya. Here in the northern zoos and aquariums many native animal species and subspecies are kept and often successfully bred.     

Pseudo-random mating systems for 3-allele loci

A random mating population attains equilibrium by the Hardy-Weinberg law. By demonstrating some simple examples for 2-allele loci, Li (1988) showed that a nonrandom mating population of certain mating patterns may also attain equilibrium. He called such a type of population a pseudo-random mating population. Tai (1990), then, gave a generalized representation of these pseudo-random mating systems. In this paper the clear patterns of pseudo-random mating behavior for a 3-allele locus are derived. Both autosomal and sex-linked systems are discussed. The study of these mating patterns provides a way to understand the complicated mating system of a population, which usually is only with difficulty realized through sampled individuals from that population.     

Slit2 Inactivates GSK3β to Signal Neurite Outgrowth Inhibition

Slit molecules comprise one of the four canonical families of axon guidance cues that steer the growth cone in the developing nervous system. Apart from their role in axon pathfinding, emerging lines of evidence suggest that a wide range of cellular processes are regulated by Slit, ranging from branch formation and fasciculation during neurite outgrowth to tumor progression and to angiogenesis. However, the molecular and cellular mechanisms downstream of Slit remain largely unknown, in part, because of a lack of a readily manipulatable system that produces easily identifiable traits in response to Slit. The present study demonstrates the feasibility of using the cell line CAD as an assay system to dissect the signaling pathways triggered by Slit. Here, we show that CAD cells express receptors for Slit (Robo1 and Robo2) and that CAD cells respond to nanomolar concentrations of Slit2 by markedly decelerating the rate of process extension. Using this system, we reveal that Slit2 inactivates GSK3β and that inhibition of GSK3β is required for Slit2 to inhibit process outgrowth. Furthermore, we show that Slit2 induces GSK3β phosphorylation and inhibits neurite outgrowth in adult dorsal root ganglion neurons, validating Slit2 signaling in primary neurons. Given that CAD cells can be conveniently manipulated using standard molecular biological methods and that the process extension phenotype regulated by Slit2 can be readily traced and quantified, the use of a cell line CAD will facilitate the identification of downstream effectors and elucidation of signaling cascade triggered by Slit.     

Hyperosmolarity enhanced susceptibility to renal tubular fibrosis by modulating catabolism of type I transforming growth factor‐β receptors

Hyperosmolarity plays an essential role in the pathogenesis of diabetic tubular fibrosis. However, the mechanism of the involvement of hyperosmolarity remains unclear. In this study, mannitol was used to evaluate the effects of hyperosmolarity on a renal distal tubule cell line (MDCK). We investigated transforming growth factor‐β receptors and their downstream fibrogenic signal proteins. We show that hyperosmolarity significantly enhances the susceptibility to exogenous transforming growth factor (TGF)‐β1, as mannitol (27.5 mM) significantly enhanced the TGF‐β1‐induced increase in fibronectin levels compared with control experiments (5.5 mM). Specifically, hyperosmolarity induced tyrosine phosphorylation on TGF‐β RII at 336 residues in a time (0–24 h) and dose (5.5–38.5 mM) dependent manner. In addition, hyperosmolarity increased the level of TGF‐β RI in a dose‐ and time‐course dependent manner. These observations may be closely related to decreased catabolism of TGF‐β RI. Hyperosmolarity significantly downregulated the expression of an inhibitory Smad (Smad7), decreased the level of Smurf 1, and reduced ubiquitination of TGF‐β RI. In addition, through the use of cycloheximide and the proteasome inhibitor MG132, we showed that hyperosmolarity significantly increased the half‐life and inhibited the protein level of TGF‐β RI by polyubiquitination and proteasomal degradation. Taken together, our data suggest that hyperosmolarity enhances cellular susceptibility to renal tubular fibrosis by activating the Smad7 pathway and increasing the stability of type I TGF‐β receptors by retarding proteasomal degradation of TGF‐β RI. This study clarifies the mechanism underlying hyperosmotic‐induced renal fibrosis in renal distal tubule cells. J. Cell. Biochem. 109: 663–671, 2010. © 2010 Wiley‐Liss, Inc.     

Activation of Ni Particles into Single Ni–N Atoms for Efficient Electrochemical Reduction of CO2

Electrochemical reduction of carbon dioxide (CO₂) to fuels and value‐added industrial chemicals is a promising strategy for keeping a healthy balance between energy supply and net carbon emissions. Here, the facile transformation of residual Ni particle catalysts in carbon nanotubes into thermally stable single Ni atoms with a possible NiN₃ moiety is reported, surrounded with a porous N‐doped carbon sheath through a one‐step nanoconfined pyrolysis strategy. These structural changes are confirmed by X‐ray absorption fine structure analysis and density functional theory (DFT) calculations. The dispersed Ni single atoms facilitate highly efficient electrocatalytic CO₂ reduction at low overpotentials to yield CO, providing a CO faradaic efficiency exceeding 90%, turnover frequency approaching 12 000 h^?1, and metal mass activity reaching about 10 600 mA mg^?1, outperforming current state‐of‐the‐art single atom catalysts for CO₂ reduction to CO. DFT calculations suggest that the Ni@N₃ (pyrrolic) site favors *COOH formation with lower free energy than Ni@N₄, in addition to exothermic CO desorption, hence enhancing electrocatalytic CO₂ conversion. This finding provides a simple, scalable, and promising route for the preparation of low‐cost, abundant, and highly active single atom catalysts, benefiting future practical CO₂ electrolysis.