Escherichia coli K-12 mutants that allow transport of maltose via the beta-galactoside transport system.

We have isolated mutants of Escherichia coli that have an altered beta-galactoside transport system. This altered transport system is able to transport a sugar, maltose, that the wild-type beta-galactoside transport system is unable to transport. The mutation that alters the specificity of the transport system is in the lacY gene, and we refer to the allele as lacYmal. The lacYmal allele was detected originally in strains in which the lac genes were fused to the malF gene. Thus, as a result of gene fusion and isolation of the lacYmal mutation, a new transport system was evolved with regulatory properties and specificity similar to those of the original maltose transport system. Maltose transport via the lacYmal gene product is independent of all of the normal maltose transport system components. The altered transport system shows a higher affinity than the wild-type transport system for two normal substrates of the beta-galactoside transport system, thiomethyl-beta-D-galactoside and o-nitrophenyl-beta-D-galactoside.     

Fine structure of the body wall,nervous system,and digestive tract in the Lobatocerebridae Rieger and the organization of the gliointerstitial system in Annelida

The new family Lobatocerebridae, Rieger, contains a group of turbellariomorph worms in the annelid line of evolution. The fine structural organization of the body wall, the digestive tract, and parts of the central and peripheral nervous system are described and the findings are discussed in light of general invertebrate cytology. The epidermis and gastrodermis contain a basal granule cell system which is structurally very similar to the neuroglia cell system of the nervous system. The continuity of the neuroglia cell system, and the epidermal basal granule cell system and the basal granule cell system in the digestive epithelia suggests the existence of a single glial-basal granule cell system, similar to the gliointerstitial cell system first recognized in the Mollusca (see Nicaise, '73). The Annelida may show a dual (ectodermal and mesodermal) origin of such a gliointerstitial cell system as suggested by similarities in the epidermal basal cell system in the Oligochaeta and of certain epidermal and gastrodermal cells in polychaete regeneration with neuroglia in the Annelida. The structural similarity of neuroglia and basal granule cells in Lobatocerebridae may be the result of similarity in the formation, maintenance, or regulation of the extracellular matrix.     

MHD diagnostics in the T-11M tokamak

An MHD diagnostic system for investigating the dynamics of disruption and the preceding phase of the discharge in the T-11M tokamak is described. This system makes it possible to study the structure of magnetic fluctuations in the plasma column. The diagnostic system includes a set of magnetic pick-up loops (Mirnov coils) arranged in several poloidal cross sections of the tokamak, a data acquisition system that provides synchronous recording of Mirnov coil signals, a synchronization system for triggering the data acquisition system during a disruption, and a system for processing and representation of the experimental data on magnetic fluctuations in the plasma column. Examples of how the MHD diagnostic system operates in the T-11M tokamak are presented.     

Co-expression of a prophage system and a plasmid system in Bacillus subtilis

A dual expression system for overexpressing two proteins by a single cell strain has been developed in Bacillus subtilis. This dual expression system combines the phi105MU331 prophage system and a plasmid system within a single cell. Protein expression by the prophage system is heat inducible, while that of the plasmid system is constitutive. Three candidate genes, BPN, BT, and amyE, all of Bacillus origin, were used as test models. Seven strains (BPN, BT, AMY, BS168K, MU331K, BPNK, and BTK) were constructed to investigate the influences of the prophage system and the plasmid system on each other, and to compare the efficiency of the individual expression systems with that of the dual expression system. Individually, the yield of the plasmid system is higher than that of the prophage system, which could be attributed to the constitutive nature of the expression of the plasmid system. Nonetheless, for the dual expression strains, the expression of two enzymes in a single fermentation run can reduce costs in facilities, manpower, and utilities. Fed-batch fermentation of BPNK strains confirmed the feasibility of applying this dual expression system in industrial-scale production.     

Comparative assessment of the resistance of the unstirred water layer to solute transport between two different intestinal perfusion systems

The resistance of the unstirred water layer to solute transport was estimated in two different intestinal single-pass perfusion systems for a comparative study, using D-glucose as a model compound. One is a well established perfusion system in anesthetized rats as a standard (system A). The other is the one in unanesthetized rats for comparison (system B). It was demonstrated that in system B as well as in system A the resistance of the unstirred water layer to D-glucose transport should be taken into account and this resistance, accordingly, the effective thickness of the unstirred water layer (delta) which is assumed to be in proportion to its resistance, could be described as a function of the perfusion rate by using a film model. The delta decreased with increasing perfusion rate and was larger in system A than in system B at each perfusion rate; 785 microns in system A versus 319 microns in system B at the perfusion rate of 0.16 ml/min and 337 microns versus 184 micron at that of 2.95 ml/min. Thus in system B the effective thickness, accordingly, the resistance, of the unstirred water layer was reduced to about 50% of that in system A, but the resistance of the unstirred water layer could still account for 85% of the total resistance at the maximum as far as D-glucose absorption was concerned, while 93% in system A. These results suggest that, compared with perfusion experiments in anesthetized rats (system A), the resistance of the unstirred water layer is reduced but cannot be left out of consideration even if perfusion experiments are performed in unanesthetized rats (system B). And the lower resistance of the unstirred water layer in system B was attributed to a turbulent flow in contrary to a laminar flow in system A.     

Granulation and immobilisation of methanogenic and sulfate-reducing bacteria in high-rate anaerobic reactors

The formation of anaerobic granular sludge on a sulfate-containing waste-water was studied in up-flow anaerobic sludge blanket reactors. Three systems were examined: a sulfidogenic system, a methanogenic system and a mixed sulfidogenic/methanogenic system. No significant granulation was observed in the sulfidogenic system. For the methanogenic and the mixed methanogenic/sulfidogenic system granulation proceeded well, and no significant difference in the granule diameter could be detected. In the three systems studied, different types of sludge developed. A (mainly) methanogenic granular sludge was developed in the methanogenic system, a (more) sulfate-reducing granular sludge was developed in the mixed methanogenic/sulfidogenic system, and a flocculant sulfate-reducing sludge was developed in the sulfidogenic system. Correspondence to: A Visser     

Characterization of a transport system for anionic amino acids in human fibroblast lysosomes

L-Aspartate and L-glutamate are transported into human fibroblast lysosomes by a single, low Km, Na(+)-independent transport system, which has been provisionally named lysosomal system d. This system resembles the Na(+)-dependent plasma membrane system chi-AG, although these differences have been observed: (1) lysosomal system d recognizes the D- as well as the L-isomers of both aspartate and glutamate, whereas only for aspartate is the D-isomer recognized by system chi-AG; (2) the anion L-homocysteate is not accepted by system chi-AG, but is an effective inhibitor of lysosomal system d; (3) N-methyl, alpha-methyl, and omega-hydroxamate derivatives of both aspartate and glutamate inhibit lysosomal system d, but only the aspartate derivatives are accepted by system chi-AG; (4) lysosomal system d shows a preference for the substrate amino group in the alpha-position, a preference not seen for system chi-AG. These points imply differences at the two recognition sites with respect to substrate length, size, and rotation, with the lysosomal site generally being the less restrictive.     

Optimal control mode of a biochemical feedback system

An optimal feedback system for constant-value control of biochemical reaction system was investigated by computer simulations. A feedback system containing a cyclic enzyme system where two enzyme types share a substrate in a cyclic manner, was found to be the most reliable one. This feedback system has a capability to keep the stationary value of the end product at a desired level against not only exogenous substrate supply but also endogenous parametric disturbances. The cyclic enzyme system installed as a control element of this feedback system played the role of comparator in this feedback system. The control mode of this feedback system was in good agreement with that of a system established by means of optimization technique based on the maximum principle. Also bang - bang control could be performed in this biochemical feedback system as well as in electrical one.     

A human motor control perspective to multiple manipulator modelling

This paper describes the aspects involved in modelling a multi-robot system from a human motor control perspective. The human motor control system has a hierarchical and decentralised structure, and building a control system for a multi-robot system that attains human features would require a decomposable model. Decomposition of a complex robotic system is difficult due to the interactions between the subsystems, so these have to be first separated before the system is modelled. The proposed method of separating the interconnections is applied with the aid of fuzzy modelling to derive a fully decomposable model of two manipulator robots handling a common object.     

Difference in transport of leucine in attached and suspended 3T3 cells.

3T3 cells in subconfluent culture take up leucine through a transport system which has a relatively high affinity for leucine (M system). When the culture becomes confluent, the M system is turned off and leucine is transported by another system which has a low affinity for leucine (S system). The M system is reactivated by transferring the cells into subconfluent cultures. In suspension cultures 3T3 cells, initiated from confluent cultures, the M system is not activated and leucine is transported by the S system. In cells suspended from subconfluent culture, the M system continues to operate at a high level for four hours and then is gradually turned off. Tumor virus transformed 3T3 cells (SV3T3 and Py3T3) grow quite well in suspension culture and transport leucine both in monolayer and suspension through a high affinity system, with a high Vmax value. A derivative of 3T3, 3T3/41, which grows in suspension much more slowly than tumor virus transformed 3T3 cells, also takes up leucine through a high affinity transport system both in monolayer and suspension but its Vmax value is lower than that of the transformed cells.     

A Switching Controller System for a Wheeled Mobile Robot

A wheeled mobile mechanism with a passive and/or active linkage mechanism for rough terrain environment is developed and evaluated. The wheeled mobile mechanism which has high mobility in rough terrain needs sophisticated system to adapt various environments.We focus on the development of a switching controller system for wheeled mobile robots in rough terrain. This system consists of two sub-systems: an environment recognition system using link angles and an adaptive control system. In the environment recognition system, we introduce a Self-Organizing Map (SOM) for clustering link angles. In the adaptive controllers, we introduce neural networks to calculate the inverse model of the wheeled mobile robot.The environment recognition system can recognize the environment in which the robot travels, and the adjustable controllers are tuned by experimental results for each environment. The dual sub-system switching controller system is experimentally evaluated. The system recognizes its environment and adapts by switching the adjustable controllers. This system demonstrates superior performance to a well-tuned single PID controller.     

Self-sustained pH oscillations in a compartmentalized enzyme reactor system

This work represents our continued effort toward fulfilling the need to discover a model system for experimental investigations of temporal oscillations in an enzyme-membrane system. In this paper, the regions in the parameter space where self-sustained pH oscillations can be induced for a compartmentalized enzyme reactor system, which consists of a well-stirred reactor, a reservoir and a membrane containing no enzyme, were determined via numerical simulation with two proteolytic enzymes: papain (EC 3.4.22.2) and alpha-chymotrypsin (EC 3.4.21.1). The sizes of the regions were qualitatively compared with those associated with enzymic membrane system. As a result, we found that the possibility of experimentally observing self-sustained oscillations in the compartmentalized papain reactor system, as well as in the papain-membrane system, is high. However, self-sustained pH oscillations are less likely in the compartmentalized alpha-chymotrypsin reactor system than in the alpha-chymotrypsin-membrane system.     

On the role of phospholipids in the reconstituted cytochrome P-450 system. A model study using dilauroyl and distearoyl glycerophosphocholine

The difference in pentoxyresorufin O-dealkylating activity observed in a reconstituted system containing dilauroylglycerophosphocholine (Lau2GroPCho) or distearoylglycerophosphocholine (Ste2GroPCho) was used as a model to study the role of phospholipids in the reconstituted cytochrome P-450b (IIB1) system. The hypotheses proposed in the literature for the role of phospholipids in the reconstituted cytochrome P-450 system, mainly based on the comparison of systems without phospholipid and with Lau2GroPCho, were either validated or shown to be unlikely when tested by comparing reconstituted systems with different phosphatidylcholines. The higher activity in the Lau2GroPCho system as compared to the Ste2GroPCho system cannot be ascribed to (a) an increased affinity of cytochrome P-450 for the NADPH-cytochrome reductase in the Lau2GroPCho system, also not to (b) a Lau2GroPCho-dependent dissociation of protein multimers, nor to (c) a change in the spin state of the heme. We found a different apparent Km for pentoxyresorufin in the Lau2GroPCho system compared with the Ste2GroPCho system. Furthermore, we found a difference between the cytochrome P-450b tryptophan fluorescence polarization of the Lau2GroPCho system and the Ste2GroPCho system as well as with a system without phosphatidylcholine. From these observations it is concluded that the higher activity of the Lau2GroPCho system compared with the Ste2GroPCho system or with a system without additional phosphatidylcholine may at least in part be caused by a difference in the conformation of the cytochrome P-450 molecules in these systems. Furthermore, the different effects of both phosphatidylcholines on the Km and V for pentoxyresorufin not only suggest a role of phospholipids in the binding of the substrate to the active site of the cytochrome P-450 molecule, but also on the efficiency of electron transfer from NADPH-cytochrome reductase to cytochrome P-450.     

Evaluation of Radiometric System for Detecting Bacteremia

An automated radiometric system (BACTEC, Johnston Laboratories) for detection of bacteremia was evaluated in parallel with a standard blood culture system in use in our laboratory. Of 1,445 blood cultures from 484 patients with possible bacteremia, 106 sets of cultures (excluding 39 presumed contaminated), representing 56 patients, were positive by both methods. The conventional system yielded 85 positive cultures from 48 patients, whereas the BACTEC system yielded 84 positive cultures from 43 patients. The BACTEC system failed to detect 22 cultures that were positive in the conventional system, and the conventional system failed to detect 21 cultures that were positive in the BACTEC system. The detection efficiency was generally equivalent in the two systems except for the lower detection rates of anaerobes and Enterobacter aerogenes by the BACTEC system and the lower detection rates of Torulopsis glabrata and, possibly, Pseudomonas sp. (group IVD) in the conventional system. The BACTEC system had a slight advantage over the conventional system in the time interval to detection of positivity. Approximately 20% of the positive cultures detected by the BACTEC system were detected on the first day of incubation compared with 7% by the conventional system. The recovery rates and detection times of anaerobes were less efficient by the BACTEC system than by the conventional system. It does not appear that the radiometric method has much advantage over available conventional methods.     

Glucose transport by mixed ruminal bacteria from a cow.

The glucose transport of mixed ruminal bacteria harvested from a holstein cow fed 5.0 kg of Italian ryegrass and 1.5 kg of flaked corn a day was investigated. The Eadie-Hofstee plot characterized two transport systems: a high-affinity, low-velocity system and a low-affinity, high-velocity system. The former system (K(m) = 16 microM; Vmax = 2.2 nmol/min/mg of protein) is considered dominant under this feeding condition based on the glucose concentration in the rumen (< 1 mM). In light of the facts that the protonophore SF6847 and the lipophilic triphenylmethyl phosphonium ion had no effect on the high-affinity system and an artificially generated proton gradient and electrical potential across the cell membrane did not increase glucose transport, a proton motive force is not be involved in the system. On the other hand, from the facts that chlorhexidine inhibited about 90% of the high-affinity system while iodoacetate showed no significant effect, and a high phosphoenolpyruvate-dependent phosphorylation of glucose was actually shown, the phosphoenolpyruvate-dependent phosphotransferase system is considered the main system in the high-affinity system. Moreover, as shown by the facts that harmaline inhibited about 30% of the high-affinity system and the artificially generated sodium gradient across the cell membrane significantly stimulated glucose transport, this system also includes sodium symport to some degree. The high-affinity system was sensitive to a decrease in pH (< 6.5) and was inhibited by the presence of sucrose, mannose, and fructose.     

国家重点生态功能区生态-经济-社会系统耦合协调的动态演化——以新疆阿勒泰地区为例

探究国家重点生态功能区生态-经济-社会系统耦合协调的演化特征,对实现可持续发展目标、建设美丽中国有重要意义。运用耦合协调模型、剪刀差方法、耦合度模型以及VAR模型对2007—2019年阿勒泰地区EES系统耦合协调度的动态演化进行分析。结果表明：(1)2007—2019年阿勒泰地区整体耦合协调度呈现不断上升趋势,由2007年的中度失调(0.271)向2019年的轻度失调(0.371)演进,生态系统与经济系统、社会系统的发展水平呈现“X”型变动趋势,经济系统与社会系统持续上升,而生态系统略微下降。(2)经济系统与社会系统、生态系统与社会系统演化速率的剪刀差在2014年以后出现较大幅度的波动,而生态系统与经济系统演化速率的剪刀差趋于稳定态势,生态系统与经济系统、经济系统与社会系统的耦合度呈现出由无序到有序的初始过渡。(3)由脉冲响应函数及方差分解可得,阿勒泰地区EES系统耦合协调水平的提升是三系统共同作用的结果,前期主要由社会系统、经济系统驱动,后期主要依赖生态系统的改善。研究表明：国家重点生态功能区应着重减轻经济系统对社会系统和生态系统的胁迫强度,进而实现三者的协同发展。     

Functional changes in cation-preferring amino acid transport during development of preimplantation mouse conceptuses

In a previous study, a Na(+)-independent, cation-preferring amino acid transport system was detected in preimplantation mouse blastocysts. The system resisted Na(+)-dependent inhibition by homoserine and so resembled the lysosomal system c more than it resembled the plasmalemmal system y+. We now report the presence of a cation-preferring system in unfertilized and fertilized eggs and cleavage-state conceptuses which also resists Na(+)-dependent inhibition by homoserine. The systems in 1-cell conceptuses and blastocysts are, however, insensitive to changes in pH in the interval of 6.0 to 8.0 and, thus, different from the pH-sensitive system c. Moreover, the relative strengths of the interactions of a variety of basic amino acids with the systems in conceptuses do not correspond well with the relative strengths of their interactions with either system c or system y+. Similarly, the system in 1-cell conceptuses can be distinguished from the system in blastocysts because L-arginine interacts about equally well with each of these systems, whereas the system in 1-cell conceptuses is inhibited more strongly than the system in blastocysts by most other basic amino acids. In addition, inhibition of the system in 1-cell conceptuses by some basic amino acids is Na(+)-stimulated, whereas Na+ does not affect inhibition of the system in blastocysts. Finally, L-tryptophan inhibits the system in blastocysts better than L-histidine or D-arginine do, but the reverse is true for the system in 1-cell conceptuses. Therefore, the relative activities of at least two forms of a novel, cation preferring amino acid transport process change during development of blastocysts from fertilized eggs. For convenience, the forms of the cation-preferring transport processes that seem to predominate at the 1-cell and blastocysts stages are provisionally designated systems b+1 and b+2, respectively, although these two systems need not represent entirely different gene products.     

Recombinant cytochromes c biogenesis systems I and II and analysis of haem delivery pathways in Escherichia coli

Genetic analysis has indicated that the system II pathway for c-type cytochrome biogenesis in Bordetella pertussis requires at least four biogenesis proteins (CcsB, CcsA, DsbD and CcsX). In this study, the eight genes (ccmA-H) associated with the system I pathway in Escherichia coli were deleted. Using B. pertussis cytochrome c4 as a reporter for cytochromes c assembly, it is demonstrated that a single fused ccsBA polypeptide can replace the function of the eight system I genes in E. coli. Thus, the CcsB and CcsA membrane complex of system II is likely to possess the haem delivery and periplasmic cytochrome c-haem ligation functions. Using recombinant system II and system I, both under control of IPTG, we have begun to study the capabilities and characteristics of each system in the same organism (E. coli). The ferrochelatase inhibitor N-methylprotoporphyrin was used to modulate haem levels in vivo and it is shown that system I can use endogenous haem at much lower levels than system II. Additionally, while system I encodes a covalently bound haem chaperone (holo-CcmE), no covalent intermediate has been found in system II. It is shown that this allows system I to use holo-CcmE as a haem reservoir, a capability system II does not possess.     

A STUDY OF THE T SYSTEM IN RAT HEART

The technique of extracellular space tracing with horseradish peroxidase is adapted for labeling the transverse tubular system (T system) in rat heart. In rat ventricular muscle the T system shows extensive branching and remarkable tortuosity. The T system can only be defined operationally, since it does not display specific morphological features throughout its entire structure. Owing to branching of the T system, a sizable proportion of the apposition between the T system and L system (or closed system) occurs at the level of longitudinal branches of the T system and is not restricted to the Z line region. The regions of apposition between the T system and L system are analyzed in rat ventricular muscle and skeletal muscle (diaphragm) and compared with the intercellular tight junctions (nexuses) of heart muscle by the use of a photometric method. The over-all thickness of the nexus is significantly smaller than that of T-L junctions in both cardiac and skeletal muscles. The thickness of the membranes of the T and L systems are not significantly different in the two muscles, but the gap between both membranes is larger in the heart. In atrial muscle the following two types of cells are found: (a) those cells with a well-developed T system in which the tubular diameter is quite uniform and the orientation predominantly longitudinal and, (b) cells with no T system, but with a well-developed L system. Atrial cells possessing a T system are richly provided with specific granules and show little micropinocytotic activity, whereas cells devoid of T system show intense micropinocytotic activity and few specific granules. The possible functional implications of these findings are discussed.