Benefits of the epilepsy specialist nurses (ESN) role, standardized practices and education around the world.

Epilepsy, often considered as a stigmatizing disease, affects 65 million people worldwide and is frequently associated with comorbidities that increase both direct and indirect costs. The degree of impact on quality of life and the cost of care differs depending on the social and health care organizations in place, political, medico-economic and/or socio-cultural contexts. Across the globe, healthcare is provided by nurses in primary care, urgent or emergency care, and within specialized domains of practice. In Epilepsy the global care could be enhanced by developing standardized nursing education in close collaboration with other caregivers. The impact of epilepsy nursing care has been documented in some developed countries, but the diversity of nursing practices and professional education of nurses raise difficulties in generalizing these findings. Specialized education in epilepsy will improve access, treatment and ultimately the quality of life of patients.

Selective amygdalohippocampectomy: surgical outcome in children versus adults.

INTRODUCTION: The objective of the study was to review our experience with selective amygdalohippocampectomy (SAH) in children and adults with intractable temporal lobe epilepsy. METHODS: A retrospective case series was used in the setting of a tertiary care hospital which provides epilepsy care to both children and adults. All patients underwent a selective amygdalohippocampectomy procedure and had at least one year of follow-up. Adults and children were divided into two groups and the data was compared between children and adults. RESULTS: Twenty three patients, 9 children and 14 adults were studied. Age of surgery varied from 6 to 58 years. Surgical outcome was variable between the two groups. Amongst the children, three patients (33%) were seizure-free (Engel Class I), two patients (22%) had rare seizures (Engel Class II), one patient (11%) had a worthwhile decrease in seizures (Engel class III) and three patients (32%) had refractory seizures that required re-operation with an anterior temporal lobectomy. This differed from the adults, who all had a good outcome. Ten patients (71%) were seizure-free (Engel Class I) and the remainder (29%) had rare seizures (Engel Class II). CONCLUSION: Selective amygdalohippocampectomy can lead to excellent seizure surgical outcome in adults with refractory temporal lobe epilepsy. However, preliminary results show less favorable results in children. The difference is probably related to the different pathology between the two groups. Anterior temporal lobe resection may prove to be a more successful operation than SAH in children with intractable temporal lobe epilepsy.

Polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha cells: a computer simulation.

Computer simulation of polyhydroxyalkanoate (PHA) granule formation in vivo could help to design strategies to optimize the fermentation process and achieve higher yields of PHA. It could also suggest biotechnological approaches to control the granule size and molecular weight of the polymer. A computer program simulating the formation of PHA granules inside a Ralstonia eutropha cell was developed, based on published experimental data. The results are applicable to R. eutropha cells or other microorganisms and transgenic plants, where polyhydroxybutyrate production is made possible by heterologous expression systems. The simulation starts at the outset of the PHA accumulation phase when the cells are small and contain no PHA granules. In the presence of abundant glucose, the cell responds to phosphorus limitation by producing 3-hydroxybutyryl-CoA which undergoes polymerization on the few PHA synthase molecules present in the cytoplasm. The amphiphilic PHA synthase-PHA complex attracts additional PHA synthase molecules and granules begin to grow from these initiation sites. Phosphorus limitation and the appearance of PHA in the cytoplasm also stimulate production of phasin molecules that attach themselves to the growing granules. As the granules grow bigger, they begin to touch each other and move to optimize their packing. The phasin coat prevents the granules from coalescing. The size of the cell increases and its prolate ellipsoid shape becomes closer to spherical. The accumulation process stops either when the supply of glucose is exhausted or when the granules become tightly packed within the cell, so that access to their surface is limited. All important variables, such as cell dimensions, granule size, counts of granule-associated molecules, PHA yield, degree of polymerization of the PHA molecules, etc., are recorded in real time during the simulation. Examples of virtual experiments with the cell and their results are shown.

The role of phasins in the morphogenesis of poly(3-hydroxybutyrate) granules.

Recent developments in the understanding of the structure of polyhydroxyalkanoate, PHA, granules in bacteria are documented in the literature and point to the role of structural proteins, phasins, in granule formation and stabilization. We have previously conceived a computer program which successfully simulates granule formation in vitro, in the absence of phasins. Now we are extending the computer model to a more complex system, including phasins, to quantify their anticipated effect on the granule properties. The simulation enabled us to propose real experiments to test the validity of the model and provide a framework for a better understanding of PHA granule formation in vivo.

Amino acid sequence and thermostability of xylanase A from Schizophyllum commune.

The amino acid sequence (197 residues) of xylanase A from the fungus, Schizophyllum commune, was determined by automated analysis of peptides from proteolytic and acid cleavage. The sequence is similar to two Trichoderma xylanases (approximately 56% identical amino acids), but also shows at least 40% identities with xylanases from Bacillus subtilis, B. pumilus and B. circulans. The conserved regions of the enzyme contain only two glutamic acid residues which implicates their possible involvement in catalysis. The disulfide bond in xylanase A is not conserved in this family. In spite of this, the B. subtilis xylanase was found to be more thermostable than xylanase A.

Manganese Peroxidase, Produced by Trametes versicolor during Pulp Bleaching, Demethylates and Delignifies Kraft Pulp.

Previous work has shown that Trametes (Coriolus) versicolor bleaches kraft pulp brownstock with the concomitant release of methanol. In this work, the fungus is shown to produce both laccase and manganese peroxidase (MnP) but not lignin peroxidase during pulp bleaching. MnP production was enhanced by the presence of pulp and/or Mn(II) ions. The maximum level of secreted MnP was coincident with the maximum rate of fungal bleaching. Culture filtrates isolated from bleaching cultures produced Mn(II)- and hydrogen peroxide-dependent pulp demethylation and delignification. Laccase and MnP were separated by ion-exchange chromatography. Purified MnP alone produced most of the demethylation and delignification exhibited by the culture filtrates. On the basis of the methanol released and the total and phenolic methoxyl contents of the pulp, it appears that MnP shows a preference for the oxidation of phenolic lignin substructures. The extensive increase in brightness observed in the fungus-treated pulp was not found with MnP alone. Therefore, either the MnP effect must be optimized or other enzymes or compounds from the fungus are also required for brightening.

Critical evaluation of comparative model building of Streptomyces griseus trypsin.

The X-ray crystal structure of Streptomyces griseus trypsin has been solved and refined at 1.7-A resolution. The structure of this protein had been predicted in two models on the basis of its expected homology to structures of bovine trypsin and other pancreatic serine proteases [Jurásek, L., Olafson, R.W., Johnson, P., & Smillie, L.B. (1976) Miami Winter Symp. 11, 93-123; Greer, J. (1981) J. Mol. Biol. 153, 1027-1042]. An evaluation of these models in light of the known structure demonstrates the effect of several sources of error on such comparative model building. The objective of comparative model building is often to explain substrate specificity, or to suggest potential highly specific drugs. The unique parts of modeled proteins that are most important for such purposes are, however, the most poorly determined by the model-building procedure.

Peroxidase-catalyzed color removal from bleach plant effluent.

Effluent from the caustic extraction stage of a bleach plant is highly colored due to the presence of dissolved products from lignin chlorination and oxidation. Color removal from the effluent by hydrogen peroxide at neutral pH was catalyzed by addition of horseradish peroxidase. The catalysis with peroxidase (20 mg/L) was observed over a wide range of peroxide concentrations (0.1mM-500mM), but the largest effect was between 1mM and 100mM. The pH optimum for catalysis was around 5.0, while the basal rate of noncatalyzed peroxide color removal simply increased with pH within the range tested (3-10). Peroxidase catalysis at pH 7.6 reached a maximum at 40 degrees C in 4 h assays with 10mM peroxide, and disappeared above 60 degrees C. Compared with mycelial color removal by Coriolus versicolor, the rate of color removal by peroxide plus peroxidase was initially faster (first 4 h), but the extent of color removal after 48 h was higher with the fungal treatment. Further addition of peroxidase to the enzyme-treated effluent did not produce additional catalysis. Thus, the peroxide/peroxidase system did not fully represent the metabolic route used by the fungus.

A fungal cellulase shows sequence homology with the active site of hen egg-white lysozyme.

The N-terminal amino acid sequence of an endo-beta-1,4-glucanase from the cellulase complex of the white-rot fungus Schizophyllum commune has been determined. The sequence from Glu-33 to Tyr-51 was homologous with the active site sequences of various hen egg-white type lysozymes, including lysozyme catalytic residues (Glu-35, Asp-52) and substrate binding residue Asn-44. The homology offers evidence for a lysozyme-type mechanism in enzymic hydrolysis of cellulose.

Isolation and Characterization of a Xylanase from Bacillus subtilis.

Partial characterization of an extracellular xylanase isolated by chromatography from Bacillus subtilis gave a molecular weight of 32,000 and optimum pH and temperature of 5.0 and 50 degrees C, respectively. K(m) and V(max) values, determined with a soluble larchwood xylan, were 0.16% and 7.0 x 10 mumol min mg of enzyme respectively. The amino acid composition showed more basic amino acid residues than in a previously characterized xylanase from a white-rot fungus.

Removal of color from kraft mill wastewaters with cultures of white-rot fungi and with immobilized mycelium of Coriolus versicolor.

Screening fifteen strains of white-rot fungi for their ability to decolorize combined bleached kraft effluent showed that Coriolus versicolor in liquid culture removed over 60% of the color of the effluent within six days in the presence of sucrose. Treatment of the same effluent with this fungus, immobilized in beads of calcium alginate gel, resulted in 80% decolorization after three days in the presence of sucrose. Caustic extraction E(1) effluent was also decolorized by the immobilized fungus. Decolorization was achieved more rapidly at pH 5.0 than at pH 7.0. Recycled beads could remove color efficiently and repeatedly in the presence of air but not under anaerobic conditions.

Fungal treatment of mechanical pulps-its effect on paper properties.

Refiner mechanical pulp was biologically treated with several higher fungi in order to test their potential for increasing the strength of paper. It was among the white-rot fungi that the best results were obtained. Polyporus versicolor gave the best overall improvement in handsheet properties with no reduction in tear. The strength improvement is due to attack on lignin and to an increase in fiber flexibility as measured by water retention values and by acidic group content of the treated pulps. The brown-rot fungi had a detrimental effect on paper properties.

Induction of cellulose in Schizophyllum commune: thiocellobiose as a new inducer.

Several mono-, di, tetra-, and polysaccharides were screened for their ability to induced cellulase production by the tetrapolar hymenomycete Schizophyllum commune. Out of 21 carbohydrates screened, 4 (thiocellobiose, carboxymethylcellulose, cellobiose, and xylan) induced all three enzymes tested (carboxymethylcellulase, beta-glucosidase, and xylanase). The inducing effect increased with rising concentrations of the inducers up to a certain value, beyond which there was either a leveling off or a decrease of the enzymatic activities. The most powerful inducer, thiocellobiose, showed the highest activity at 0.5 mM. Cellobiose, carboxymethylcellulose, and xylan showed their highest activities at 1 mM and 1%, respectively. Surprisingly, sophorose did not enhance enzyme production. The enzymatic activities were monitored over a period of 24 h. Thiocelloboise elicited a response immediately after incubation, but with all other inducers there was a latency period before their effect could be measured. High-performance liquid chromatography showed no hydrolysis of thiocellobiose when incubated in the presence of S. commune extracellular enzymes.

High Production of beta-Glucosidase in Schizophyllum commune: Isolation of the Enzyme and Effect of the Culture Filtrate on Cellulose Hydrolysis.

Optimization experiments with response surface statistical analysis were performed with Schizophyllum commune to obtain high beta-glucosidase yields. The factors in the optimization experiment were the concentrations of cellulose, peptone, and KH(2)PO(4). Their optimal values were 3.2, 3.0, and 0.2 g/100 ml, respectively. Enzyme assays revealed very high beta-glucosidase (22.2 U/ml) and cellobiase (68.9 U/ml) yields. The avicelase yield was low as compared with that from Trichoderma reesei. Mixtures of S. commune and T. reesei culture filtrates caused faster and more extensive saccharification of Avicel than could be achieved by either filtrate alone. A beta-glucosidase was isolated and purified from the optimized culture filtrate of S. commune. The electrophoretic mobility of the purified beta-glucosidase indicated a molecular weight of 97,000. The amino acid composition was similar to that of beta-glucosidase from T. reesei. The acidic (aspartate and glutamate) residues or their amides or both made up approximately 20% of the protein. The NH(2)-terminal amino acid of the enzyme was histidine.

Production, characterization, and partial amino acid sequence of xylanase A from Schizophyllum commune.

Xylanase A, one of several extracellular xylanases produced by Schizophyllum commune strain Delmar when grown in submerged culture with spruce sawdust as carbon source, was purified 43-fold in 25% yield with respect to total xylanase activity. Although some polysaccharide was strongly bound to the purified enzyme, the complex could be dissociated by sodium dodecyl sulfate and appeared homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular weight of the protein, calculated from the electrophoretic mobility, was 33,000. The molecular activity of the purified xylanase A, determined with soluble larch xylan as substrate, was 1.4 X 10(5) min-1, with xylobiose and xylose as the major products. The enzyme had a pH optimum of 5.0 and a temperature optimum of 55 degrees C in 10-min assays. The acid hydrolysate of xylanase A was rich in aspartic acid and aromatic amino acids. The sequence of 27 residues at the amino terminus showed no homology with known sequences of other proteins.

Amino acid sequence of Streptomyces griseus trypsin. Cyanogen bromide fragments and complete sequence.

Information compiled by automatic Edman degradation of Streptomyces griseus trypsin coupled with previous data has permitted the assignment of the first 36 residues at the NH2 terminus of the protein. Cyanogen bromide cleavage at the three methionine residues followed or preceded by reduction and aminoethylation resulted in the production of four fragments, Cnl to Cn4, which were separated by gel filtration on Sephadex G-50 or G-75. Fragments CN4 (15 RESIDUES) AND Cn3 (5 residues) were shown to be derived from the NH2 terminus of the protein while Cn2 (47 residues and devoid of homoserine) was from the COOH terminus. The arrangement of the fragments was thus Cn4-Cn3-Cn1-Cn2. Automatic Edman degradation in the sequenator coupled with peptides derived from alpha-lytic protease and chymotryptic digestion and from the peptic and tryptic peptides previously elucidated have permitted the sequence determination of fragments Cn1 and Cn2 and therefore of the whole protein. These studies show that extensive regions of identity or similarity exist between Streptomyces griseus trypsin and bovine trypsin. These include the NH2-terminal four residues, the sequences near histidine-57 (chymotrypsinogen A numbering system), aspartic acid-102, aspartic acid-189, and serine-195, the regions of the three disulfide bridges, and the COOH-terminal end (residues 225-229) of the proteins. When aligned to maximize homology the identity of residues is 34%. This identity is increased to 54% when only those residues classified as internal by Stroud et al. (Stroud, R. M., Kay, L. M., and Dickerson, R. E. (1971) Cold Spring Harbor Symp. Quant. Biol. 36, 125) are considered. These results indicate that the folding of the polypeptide chains of the two enzymes is very similar and are in agreement with the very similar enzymic, chemical, and physical properties of the two enzymes.