Lethality of the covalent linkage between mislocalized major outer membrane lipoprotein and the peptidoglycan of Escherichia coli

The major outer membrane lipoprotein (Lpp) of Escherichia coli possesses serine at position 2, which is thought to function as the outer membrane sorting signal, and lysine at the C terminus, through which Lpp covalently associates with peptidoglycan. Arginine (R) is present before the C-terminal lysine in the wild-type Lpp (LppSK). By replacing serine (S) at position 2 with aspartate (D), the putative inner membrane sorting signal, and by deleting lysine (K) at the C terminus, Lpp mutants with a different residue at either position 2 (LppDK) or the C terminus (LppSR) or both (LppDR) were constructed. Expression of LppSR and LppDR little affected the growth of E. coli. In contrast, the number of viable cells immediately decreased when LppDK was expressed. Prolonged expression of LppDK inhibited separation of the inner and outer membranes by sucrose density gradient centrifugation, whereas short-term expression did not. Pulse-labeled LppDK and LppDR were localized in the inner membrane, indicating that the amino acid residue at position 2 functions as a sorting signal for the membrane localization of Lpp. LppDK accumulated in the inner membrane covalently associated with the peptidoglycan and thus prevented the separation of the two membranes. Globomycin, an inhibitor of lipoprotein-specific signal peptidase II, was lethal for E. coli only when Lpp possessed the C-terminal lysine. Taken together, these results indicate that the inner membrane accumulation of Lpp per se is not lethal for E. coli. Instead, a covalent linkage between the inner membrane Lpp having the C-terminal lysine and the peptidoglycan is lethal for E. coli, presumably due to the disruption of the cell surface integrity.     

Fuzzy inference and fuzzy inference processor

Fuzzy inference, a data processing method based on the fuzzy theory that has found wide use in the control field, is reviewed. Consumer electronics, which accounts for most current applications of this concept, does not require very high speeds. Although software running on a conventional microprocessor can perform these inferences, high-speed control applications require much greater speeds. A fuzzy inference date processor that operates at 200000 fuzzy logic inferences per second and features 12-b input and 16-b output resolution is described     

Pixel design of pulsed CMOS image sensor for retinal prosthesis with digital photosensitivity control

A newly designed pulse frequency modulation photosensor for use in retinal prosthesis is proposed and demonstrated. The pixel converts the intensity of incident light into biphasic current pulses at frequencies suitable for the electrical stimulation of retinal neurons. Experimental results showed that the device was sensitive over a dynamic range of input light of about 120 dB, and that photosensitivity could be varied from 0 dB to around -40 dB.     

Stability analysis of regional and national voting schemes by a continuous model

The previous discrete-model-based stability analysis of regional and national voting has been extended to a continuous-model-based analysis in the simultaneous presence of white and concentrated components of noise, reconfirming the previous conclusion that regional voting with smaller sized regions always demonstrates an improved stability over those with larger sized regions, including the national voting in its limiting case in particular. The conclusion remains valid as long as the weak distribution assumption is valid.     

Involvement of specific mechanism in plasmid DNA uptake by mouse peritoneal macrophages

Lamina-associated polypeptide (LAP)2, which directly interacts with B-type lamins and chromosomes, is an integral membrane protein specifically distributed along the inner nuclear membrane of the nuclear envelope. Multiple regions of its large nucleoplasmic domain promote this localization, including the first (residues 1-296) and the second (residues 298-409) halves of the LAP2 N terminus. The second half is involved in LAP2 association with the nuclear lamina [Furukawa, K., Panté, N., Aebi, U. & Gerace, L. (1995) EMBO J. 14, 1626-1636]. In this study to further define its role, we examined which domain of B-type lamin interacts with LAP2 by means of a binding assay with bacterially expressed proteins and a yeast two-hybrid system. We found that amino acids in the region of residues 78-258 of the lamin B1 rod domain directly bound with LAP2. The data suggest that LAP2 may modulate the assembly of nuclear lamins.     

New catalytic functions of Pd–Zn, Pd–Ga, Pd–In, Pt–Zn, Pt–Ga and Pt–In alloys in the conversions of methanol

Pd and Pt supported on ZnO, Ga₂O₃ and In₂O₃ exhibit high catalytic performance for the steam reforming of methanol, CH₃OH+H₂OCO₂+3HH₂, and the dehydrogenation of methanol to HCOOCH₃, 2CH₃OHHCOOCH₃+2HH₂. Combined results with temperature-programmed reduction (TPR) and XRD method revealed that Pd–Zn, Pd–Ga, Pd–In, Pt–Zn, Pt–Ga and Pt–In alloys were produced upon reduction. Over the catalysts having the alloy phase, the reactions proceeded selectively, whereas the catalysts having metallic phase exhibited poor selectivities.     

Alteration of substrate specificity of cholesterol oxidase from Streptomyces sp. by site-directed mutagenesis

Despite the structural similarities between cholesterol oxidasefrom Streptomyces and that from Brevibacterium, both enzymesexhibit different characteristics, such as catalytic activity,optimum pH and temperature. In attempts to define the molecularbasis of differences in catalytic activity or stability, substitutionsat six amino acid residues were introduced into cholesteroloxidase using site-directed mutagenesis of its gene. The aminoacid substitutions chosen were based on structural comparisonsof cholesterol oxidases from Streptomyces and Brevibacterium.Seven mutant enzymes were constructed with the following aminoacid substitutions: L117P, L119A, L119F, V145Q, Q286R, P357Nand S379T. All the mutant enzymes exhibited activity with theexception of that with the L117P mutation. The resulting V145Qmutant enzyme has low activities for all substrates examinedand the S379T mutant enzyme showed markedly altered substratespecificity compared with the wild-type enzyme. To evaluatethe role of V145 and S379 residues in the reaction, mutantswith two additional substitutions in V145 and four in S379 wereconstructed. The mutant enzymes created by the replacement ofV145 by Asp and Glu had much lower catalytic efficiency forcholesterol and pregnenolone as substrates than the wild-typeenzyme. From previous studies and this study, the V145 residueseems to be important for the stability and substrate bindingof the cholesterol oxidase. In contrast, the catalytic efficiencies(k_cat/K_m) of the S379T mutant enzyme for cholesterol and pregnenolonewere 1.8- and 6.0-fold higher, respectively, than those of thewild-type enzyme. The enhanced catalytic efficiency of the S379Tmutant enzyme for pregnenolone was due to a slightly high k_catvalue and a low K_m value. These findings will provide severalideas for the design of more powerful enzymes that can be appliedto clinical determination of serum cholesterol levels and assterol probes.     

High-temperature oxidation of pure titanium in CO2 and Ar-10%CO2 atmospheres

The oxidation behavior of pure titanium has been investigated in the temperature range of 1000 K to 1300 K in CO₂ or Ar-10%CO₂. Optical microscopy, electron probe microanalyses, and X-ray measurements on the oxide scales formed during oxidation indicate that their structures are nearly independent of temperature and the corrosion atmosphere. The scales consisted of two layers, an external one and an internal one, having a rutile (TiO₂) structure. The parabolic rate law was confirmed for growth of the external scale and the permeation depth of oxygen in titanium with apparent activation energies of 266 and 226 kJ/mol, respectively. The rate-determining diffusion species in the oxidation processes are discussed.     

Managing a Very Connected World: A Report on APNOMS2015

This article presents a report on APNOMS2015, which was held August 19–21, 2015 in Busan, Korea. The theme of APNOMS2015 was “Managing a Very Connected World.”     

Mean–variance relationship of the number of flows in traffic aggregation and its application to traffic management

We consider the mean–variance relationship of the number of flows in traffic aggregation, where flows are divided into several groups randomly, based on a predefined flow aggregation index, such as source IP address. We first derive a quadratic relationship between the mean and the variance of the number of flows belonging to a randomly chosen traffic aggregation group. Note here that the result is applicable to sampled flows obtained through packet sampling. We then show that our analytically derived mean–variance relationship fits well those in actual packet trace data sets. Next, we present two applications of the mean–variance relationship to traffic management. One is an application to detecting network anomalies through monitoring a time series of traffic. Using the mean–variance relationship, we determine the traffic aggregation level in traffic monitoring so that it meets two predefined requirements on false positive and false negative ratios simultaneously. The other is an application to load balancing among network equipments that require per-flow management. We utilize the mean–variance relationship for estimating the processing capability required in each network equipment.