A Novel Robot System Integrating Biological and Mechanical Intelligence Based on Dissociated Neural Network-Controlled Closed-Loop Environment.

We propose the architecture of a novel robot system merging biological and artificial intelligence based on a neural controller connected to an external agent. We initially built a framework that connected the dissociated neural network to a mobile robot system to implement a realistic vehicle. The mobile robot system characterized by a camera and two-wheeled robot was designed to execute the target-searching task. We modified a software architecture and developed a home-made stimulation generator to build a bi-directional connection between the biological and the artificial components via simple binomial coding/decoding schemes. In this paper, we utilized a specific hierarchical dissociated neural network for the first time as the neural controller. Based on our work, neural cultures were successfully employed to control an artificial agent resulting in high performance. Surprisingly, under the tetanus stimulus training, the robot performed better and better with the increasement of training cycle because of the short-term plasticity of neural network (a kind of reinforced learning). Comparing to the work previously reported, we adopted an effective experimental proposal (i.e. increasing the training cycle) to make sure of the occurrence of the short-term plasticity, and preliminarily demonstrated that the improvement of the robot's performance could be caused independently by the plasticity development of dissociated neural network. This new framework may provide some possible solutions for the learning abilities of intelligent robots by the engineering application of the plasticity processing of neural networks, also for the development of theoretical inspiration for the next generation neuro-prostheses on the basis of the bi-directional exchange of information within the hierarchical neural networks.

Production and purification of ovine anti-tetanus antibody.

We used the ovine as bioreactor for the production and optimization of anti-tetanus toxin antibody. Four female sheep were immunized with human tetanus vaccine (TT-alum) every two weeks for 16 weeks, after which serum was collected and its titer was estimated by ELISA. The highest titer obtained was 39,000 IU ml-1. To optimize a purification protocol for ovine anti-tetanus toxin, we used four procedures; weak anion (DEAE-Sephadex), weak cation (CM-Sephadex), ammonium sulfate precipitation alone or in combination with caprylic acid. Fifty percent saturation with ammonium sulfate combined with caprylic acid gave us the highest yield of protein with specific activity and the purest Fab product.

Tetanus toxin-induced protein kinase C activation and elevated serotonin levels in the perinatal rat brain.

A single intracerebral injection of tetanus toxin (TeTox) is able to produce a time-dependent translocation of Ca2(+)-phosphatidylserine-dependent protein kinase C (PKC) in close-to-term rat brain. TeTox-triggered translocation of PKC is dose- and time-dependent, can be prevented by tetanus antitoxin, and does not occur upon administration of toxin fragments B and C. TeTox-triggered PKC translocation is accompanied by a time-dependent increase in brain serotonin (5-HT). Increase of brain 5-HT is independent of monoamine oxidase inhibition by pargyline. Phorbol ester and TeTox cause a significant increase in serotonin while H-7, a kinase inhibitor, does not affect serotonin levels but abolishes the effect of TeTox. Gangliosides prevent TeTox-triggered 5-HT increase. The data are consistent with the possibility that TeTox acts effectively on the serotonergic innervation, presumably in conjunction with PKC to cause accumulation of serotonin.

Platelet aggregation induced by antilymphocyte serum. I. Differences between washed and unwashed platelets.

Horse antilymphocyte serum (ALS) induced active release of nucleotides and aggregation of washed human platelets. It also caused lysis of platelets in the presence of plasma. The sequence of lytic processes was studied by electron microscopy. Aggregation induced by ALS was not mediated through ADP in either washed or unwashed platelets, but was inhibited by those inhibitors of secondary platelet aggregation. In unwashed platelets heparin inhibited the cytolysis and aggregation completely, while prostaglandin E1 (PGE1) did not; in contrast, PGE1 completely inhibited the aggregation and release of nucleotides from washed platelets, while heparin did not. The possible mechanisms of ALS-induced aggregation are discussed.

At least three sequential steps are involved in the tetanus toxin-induced block of neuromuscular transmission.

Tetanus toxin causes a block of the neuromuscular transmission. The kinetic aspects of the block were studied in vitro on the mouse phrenic nerve-hemidiaphragm exposed to toxin (1 microgram/ml). 1. The toxin action on the nerve ending involves three sequential steps: binding, "translocation" and paralysis. 2. Diffusion and binding of tetanus toxin molecules to the presynaptic membrane is complete in about 60 min. The binding step is irreversible, independent of transmitter release and of the temperature. Tetanus antitoxin, however, inactivates the bound toxin molecules. 3. After a second step which is probably due to a "translocation" of the toxin molecules into or through the presynaptic membrane the antitoxin molecules are now ineffective to prevent the toxin-induced inhibition of transmitter release. This so called "translocation" step requires transmitter release and therefore depends strongly on the frequency of nerve stimulation. 4. The paralytic step does not depend on the transmitter release. It, however, depends strongly on temperature with a break in the Arrhenius-plot around 33 degrees C which suggests the involvement of a phase transition rather than of an enzymatic activity of the toxin.

[Heat stability of potency of bacterial vaccines and the titer of tetanus antitoxin of Polish production]. / Stabilnosc cieplna mocy szczepionek bakteryjnych oraz miano antytoksyny tezcowej produkcji krajowej.

Described the stability of potency vaccines (DTP, BCG) and immunoglobulins (human's and animal's) at storage and experimental temperatures. Thermal degradation rate and design of loss of potency in time have been determined by Arrhenius equation. Our results were similar to WHO data from preparations which have been made in another countries.

[Effect of tetanic anatoxin and carbonic acid level in blood on biosynthesis of antibodies in rats]. / Vliianie stolbniachnogo anatoksina i urovnia uglekisloty v krovi na obrazovanie antitel u krys.

The paper deals with dynamics of antibody formation in rats. Tetanic anatoxin independently of doses and methods of administration manifests weak immunogenic properties. A complete Freind adjuvant stimulates considerably the biosynthesis of antitetanic antibodies in the primary and secondary immune responses. Electrophoresis of tetanic anatoxin polyacrylamide gel permits isolating two protein fractions, one of them migrates in the immunoglobulin G zone and its mass constitutes 73%, the other-more low-molecular and contains 27% of protein. Dynamics of the acid-base blood state indices and of antibody biosynthesis is studied on models of chronic compensated alkalosis and metabolic acidosis in rats. It is established that during the experiment, simultaneously with an insignificant increase in the total carbonic acid concentration blood, there occurs a 25-60% increase in the titres of antitetanic antibodies as compared to the control. In rats with acidosis there occurs a simultaneously decrease in the intensity of antibody biosynthesis as compared to the control (43-54%) and in the total CO2 content in blood.

[Study of the chromosomes in the bone marrow cells of mice inoculated subsequently with various vaccines]. / Issledovanie khromosom v kletkakh kostnogo kozga myshei, privitykh posledovatel'no razlichnymi vaktsinami

Studies in the effect of a complex of inoculating preparations (poliovaccine, APDD-vaccine, smallpox vaccine, measles vaccine) on dividing cells of bone marrow in mice in line CC57Br showed that a reduction of the interval between introduction of vaccines different in the antigenic respect from 14 days to 4 days results in an increase in frequency of structural chromosomal aberrations 1-2 months after the whole course of inoculations.

Sequencing and phylogenetic analysis of neurotoxin gene from an environmental isolate of Clostridium sp.: comparison with other clostridial neurotoxins.

A Clostridium sp. isolated from intestine of decaying fish exhibited 99% sequence identity with C. tetani at 16S rRNA level. It produced a neurotoxin that was neutralized by botulinum antitoxin (A+B+E) as well as tetanus antitoxin. The gene fragments for light chain, C-terminal and N-terminal regions of the heavy chain of the toxin were amplified using three reported primer sets for tetanus neurotoxin (TeNT). The neurotoxin gene fragments were cloned in Escherichia coli and sequenced. The sequences obtained exhibited approximately 98, 99 and 98% sequence identity with reported gene sequences of TeNT/LC, TeNT/HC and TeNT/HN, respectively. The phylogenetic interrelationship between the neurotoxin gene of Clostridium sp. with previously reported gene sequences of Clostridium botulinum A to G and C. tetani was examined by analysis of differences in the nucleotide sequences. Six amino acids were substituted at four different positions in the light chain of neurotoxin from the isolate when compared with the reported closest sequence of TeNT. Of these, four were located in the beta15 motif at a solvent inaccessible, buried region of the protein molecule. One of these substitutions were on the solvent accessible surface residue of alpha1 motif, previously shown to have strong sequence conservation. A substitution of two amino acids observed in N-terminal region of heavy chain were buried residues, located in the beta21 and beta37 motifs showing variability in other related sequences. The C-terminal region responsible for binding to receptor was conserved, showing no changes in the amino acid sequence.