Origin oriented management of inverted papilloma of the frontal sinus.

BACKGROUND: Despite the great progress in endoscopic management of inverted papilloma (IP), involvement of the frontal sinus (FS) remains a challenge. METHODOLOGY: Six cases of FS IP were assessed. Extent of surgery included simple frontal recess clearance, extended frontal sinusotomy, and modified Lothrop approach. There was no need for adjuvant frontal trephination or an external osteoplastic flap. RESULTS: FS involvement was observed in 6 out of 119 cases of IP (5%). In one case, IP was originating from the FS and in four it was extending to the FS. The sixth case had a wide origin from the anterior ethmoid and FS. Complete resection of FS IP was achieved in all cases with a single incidence of CSF leak. No recurrence was identified after a follow-up period of an average of 27 months. CONCLUSIONS: FS IP originating outside FS can be delivered transnasally with or without frontal ostium widening and preserving FS mucosa and bone. Inverted papillomata originating from FS proper and those with origin from inside and outside the FS can also be resected tranasnasally after widening of the frontal ostium with removal of surrounding mucosa and drilling or curettage of underlying bone at attachment sites.

Biosynthesis of cyclodextrin glucosyltransferase by the free and immobilized cells of Bacillus cereus NRC7 in batch and continuous cultures.

AIMS: The objective of this study was to enhance the production of cyclodextrin glucanotransferase (CGTase) produced by a local isolate Bacillus cereus NRC7. METHODS AND RESULTS: In batch culture, maximal CGTase activity (69·0 U ml(-1)) was reached after 24-h incubation period. In continuous production of CGTase by the free cells of B. cereus NRC7, maximal reactor productivity (11·76 KU l(-1) h(-1)), with enzyme concentration of 49·0 U ml(-1) and specific productivity of 904·6 U per g wet cells per h, was attained at dilution rate of 0·24 h(-1), over a period of 640 h. Bacillus cereus NRC7 cells were immobilized on chitosan. The immobilization conditions with respect to matrix concentration and maximal cell loading were optimized for maximal CGTase production. In repeated batch operation, the activity of the immobilized cells was stable during ten cycles and the activity remained between 51 and 55 U ml(-1). In packed-bed reactor, the immobilized cells showed maximal productivity (27·18 KU l(-1) h(-1)) with enzyme concentration of 54·63 U ml(-1) and specific productivity of 151·89 U per g wet cells per h at dilution rate of 0·5 h(-1). The half-life of the immobilized cells was higher than 20 days. CONCLUSIONS: Continuous fermentation by the immobilized cells in packed-bed reactor is an appropriate potential technique for B. cereus NRC7 CGTase production that gave maximum productivity (27·18 KU l(-1) h(-1)), which was 9·47-, 2·31-, 12·24- and 12·94-fold higher than the free cells in batch, free cells in continuous, immobilized cells in batch and repeated batch cultures, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that evaluates CGTase productivity, in different fermentation modes, in terms of specific productivity (U per gram cells per h). In continuous fermentation by immobilized cells, maximal levels of CGTase productivity are higher than the previously reported values.

Production and properties of beta-mannanase by free and immobilized cells of Aspergillus oryzae NRRL 3488.

Seven fungi were tested for production of mannanases. The highest mannanase activities were produced by Aspergillus oryzae NRRL 3488 after 7 days in static cultures. Mannanases were induced by gum locust bean (1.0%). The highest mannanase activity was produced when a mixture of peptone, urea and ammonium sulphate was used as nitrogen source. Zn2+ or Co2+ favoured enzyme production. The immobilized cells on Ca-alginate and agar were able to produce beta-mannanase for four runs with a slight decrease in the activity. The optimum temperature for enzyme reaction was 50-55 degrees C at pH 6.0. In the absence of substrate the enzyme was thermostable retaining 75% activity for 1 h at 50 degrees C, and 68% activity for 1 h at 60 degrees C.

Effect of thymus extract on immunologic reactivity of chicken vaccinated with infectious bursal disease virus.

The effects of crude thymus extract on the immune response and protection against challenge with virulent infectious bursal disease virus (IBDV) were studied in one-day-old chick. Oral administration of thymus extract (1 ml/kg) markedly and significantly increased the total protein, albumin, globulin, Tri-iodothyronine (T3), Thyroxine (T4) and the body weight gain in one-day-old chick. In addition, it increased the total lymphocytic count over four weeks after administration. Although vaccination also increased total protein, globulin, T4 and the total lymphocytic count but it significantly decreased the body weight gain of the chick and administration of thymus extract, before, during or after vaccination markedly improved the vaccination effectiveness with significant elevation of the globulin level and body weight gain of the chick. It also prevented the decrease in the relative weights of bursa, spleen and thyroid gland which commonly prevailed during vaccination. Chicken administered thymus extract and vaccinated with infectious bursal disease (IBD) vaccine showed 100% protection against challenge with IBDV. Meanwhile the vaccinated non-thymus treated group exhibited 80% protection against IBDV challenge. These results indicate a potentiating effect of thymus extract on the immune system in baby chick. These findings are supported by ELISA results that showed a marked increase in antibody titers in thymus treated groups. Additionally, microscopical examination of the bursa and the existent lymphoid hyperplasia in thymus treated groups but not vaccinated group support our findings.

Purification and properties of three cellobiases from Aspergillus niger A20.

Three cellobiases, here called cellobiase A, B, and C, from the culture filtrate of Aspergillus niger A20, were purified by precipitation with ammonium sulphate, gel filtration through Sephadex G-75, and column chromatography of DEAE-cellulose. The purified enzymes were homogeneous on polyacrylamide disk electrophoresis. The mol wt of the purified enzymes were estimated by SDS-gel electrophoresis to be 88,000, 80,000, and 71,000 for cellobiases A, B, and C, respectively. The enzymes were active at pH 4.5 and 55-60 degrees C. The pattern of their amino acid compositions showed high contents of aspartic acid, glutamic acid, threonine, serine, and glycine. The apparent K(m) values for cellobiose were 0.9, 1.63, and 1.0 mM for cellobiases A, B, and C, respectively. Calcium ions stimulated cellobiases B and C, and Co2+ and Mg2+ ions stimulated cellobiase A. The purified enzymes hydrolyzed cellobiose and aryl-beta-D-glucosides, but they had no action on sucrose, maltose, and cellulose. The three cellobiases catalyzed transglycosylase reaction, and the major product formed from cellobiose was tetramer of glucose.

Central serotonin level-dependent changes in body temperature following administration of tryptophan to pargyline- and harmaline-pretreated rats.

1. The effect of tryptophan on body temperature was studied in rats pretreated with pargyline, an irreversible monoamine oxidase inhibitor (MAOI), and harmaline, a reversible MAOI. 2. Tryptophan (100 mg/kg IP) produced hypothermia followed by hyperthermia in pargyline-pretreated rats, and hypothermia in harmaline-pretreated rats, but tryptophan did not cause body temperature changes by itself. 3. The tryptophan-induced hypo- and hyperthermic effects, which peaked at about 1 and 6 hr after tryptophan administration, respectively, were accompanied by a significant increase in serotonin (5-HT) levels in the pargyline-pretreated rat brain (75%-138.7% and 207%-240.9% increase, respectively), and the 5-HT levels in the hyperthermic state were significantly higher than those in the hypothermic state. 4. In harmaline-pretreated rats, tryptophan also increased the central 5-HT levels (80.5%-95.5% increase) in the hypothermic state, and the effect peaked at about 1 hr after tryptophan administration. The central 5-HT levels in harmaline-pretreated rats slightly decreased at 6 hr after tryptophan administration and were significantly lower than those in the hyperthermic state in the pargyline-pretreated rats. 5. Tryptophan (100 mg/kg IP) administration decreased 5-hydroxy indole acetic acid (5-HIAA) levels, 5-HT turnover, and dopamine (DA) turnover in the brain of pargyline-pretreated rats, but these parameters were not significantly different between the hypothermic and hyperthermic states (i.e., at 1 and 6 hr after tryptophan administration, respectively). 6. These results suggest that the tryptophan-induced body temperature change depends on the different 5-HT levels in the brain and that the 5-HT level needed to induce hyperthermia is higher than that needed to induce hypothermia.

Facilitatory and inhibitory effects of harmaline on the tryptophan-induced 5-hydroxytryptamine syndrome and body temperature changes in pargyline-pretreated rats.

The effects of harmaline on tryptophan-induced 5-hydroxytryptamine (5HT) syndrome and body temperature changes in pargyline-pretreated rats were investigated. When administered i.p. 60 min after pargyline treatment (50 mg/kg, i.p.), tryptophan, at 100 mg/kg but not 10 mg/kg, induced the 5-HT syndrome. Tryptophan at 100 mg/kg also produced hypothermia followed by hyperthermia in pargyline-pretreated rats. Administration of harmaline (10 mg/kg, i.p.) 30 min after pargyline not only potentiated the 100 mg/kg tryptophan-induced 5-HT syndrome and body temperature changes, but also produced the syndrome following administration of 10 mg/kg tryptophan in pargyline-pretreated rats. In contrast, when administered 30 min before parygline, 10 mg/kg harmaline completely suppressed the syndrome and body temperature changes caused by mg/kg tryptophan. Tryptophan (100 mg/kg, i.p.) administration significantly increased 5-HT levels and decreased 5-hydroxyindole acetic and levels and 5-HT turnover in the brain of pargyline-pretreated rats. Harmaline administration 30 min after pargyline did not significantly affect the tryptophan-induced changes in 5-HT levels and 5-HT turnover, whereas when administered 30 min before pargyline, harmaline significantly blocked the effect of tryptophan. These results suggest that mechanisms underlying the inhibitory action of harmaline on the tryptophan-induced 5-HT syndrome and body temperature changes in pargyline-pretreated rats differ from those by which harmaline potentiates the effects of tryptophan.

5-HT1A and 5-HT2 receptors mediate hypo- and hyperthermic effects of tryptophan in pargyline-pretreated rats.

Mechanisms of tryptophan (a 5-HT precursor)-induced changes in body temperature were investigated in rats pretreated with pargyline, a monoamine oxidase inhibitor (MAO-I). Tryptophan (100 mg/kg, i.p.) did not affect the body temperature in rats, but it produced significant hypothermia followed by marked hyperthermia and higher mortality in the pargyline-pretreated rats. 5-HT depletion with p-chlorophenylalanine (p-CPA, 100 mg/kg/day for 3 days) significantly suppressed not only the body temperature change but also the mortality and 5-HT syndrome following tryptophan plus pargyline administration. Although propranolol (10 mg/kg, i.p.), a beta-adrenoceptor antagonist, did not alter the hypothermia caused by tryptophan in the pargyline-pretreated rats, pindolol (2 mg/kg, S.C.), a 5-HT1A receptor and beta-adrenoceptor antagonist, suppressed the hypothermia but not the hyperthermia or mortality caused by the same treatment. On the other hand, spiperone and ketanserin, 5-HT2 receptor antagonists, at doses of 3 mg/kg, potentiated the hypothermia and completely suppressed the hyperthermia and mortality caused by tryptophan in the pargyline-pretreated rats. These results suggest that tryptophan-induced hypo- and hyperthermia are mediated by 5-HT1A and 5-HT2 receptors, respectively, in the pargyline-pretreated pretreated rats.

Hypothermic effect of harmala alkaloid in rats: involvement of serotonergic mechanism.

The effect of total alkaloid extracted from Peganum harmala seeds collected in Egypt on body temperature was studied in rats. Intraperitoneal administration of the Peganum harmala extract produced significant and dose-dependent hypothermia. Similarly, harmine and harmaline, major constituents of the harmala alkaloid, lowered the body temperature. Pretreatment with p-chlorophenylalanine (100 mg/kg/day for 3 days), a 5-HT synthesis inhibitor, significantly attenuated the hypothermic effect of the total alkaloid and harmine, while it tended to block the hypothermic action of harmaline. Methysergide (2 mg/kg), a 5-HT antagonist, significantly attenuated the hypothermia induced by harmala alkaloids. Pindolol (0.05-2 mg/kg), a 5-HT1A receptor and beta-adrenoceptor antagonist, partly blocked the hypothermic effect of the harmala alkaloids in a dose-dependent manner, whereas propranolol (10 mg/kg), a beta-adrenoceptor antagonist, failed to alter it, suggesting that beta-adrenoceptor is not involved in the hypothermia caused by the alkaloids. Pretreatment with a dopamine receptor antagonist haloperidol (5 mg/kg, s.c. and 2 mg/kg, i.p. 24 and 2 h before the experiment, respectively) significantly attenuated the hypothermic effect of harmala alkaloids. Moreover, in haloperidol pretreated rats, methysergide (2 mg/kg, i.p.) and pindolol (0.05 and 2 mg/kg) completely attenuated the hypothermic effect of the alkaloids. These data suggest that harmala alkaloids produce hypothermic effect mainly through endogenous 5-HT stimulation of 5-HT1A receptor.

Utilization of water hyacinth cellulose for production of cellobiase-rich preparation by Aspergillus niger 1.

Production of a cellobiase-rich preparation by Aspergillus niger 1 was achieved using water hyacinth cellulose as the sole carbon source in the culture medium. Production of cellobiase, carboxymethylcellulase (CMC-ase) and filter paper (FP)-cellulase was favoured by controlling the pH of the culture medium during fermentation at 5.0. Sodium citrate (0.5%), sodium phytate (0.1%), Tween-80 (0.2%, v/v) and asparagine (0.07%) had stimulating effects on the productivity of cellobiase, CMC-ase and FP-cellulase. Potassium dihydrogen phosphate doubled the yield of CMC-ase but had a slight effect on FP-cellulase and cellobiase. Wheat bran had a pronounced stimulating effect on the production of cellobiase and CMC-ase. The combined effects of these stimulators resulted in an enzyme preparation rich in cellobiase and contained 18.5, 0.29 and 2.21 U/ml of cellobiase, FP-cellulase and CMC-ase, respectively. A high cellobiase/FP-cellulase ratio of 63.8:1 was thus obtained with the fungal enzyme preparation. The cellobiase activity was maximal at pH 5.0 and showed good thermostability.

Production and some properties of fibrinolytic enzymes from Fusarium oxysporum N.R.C.1.

A mixture of yeast extract and peptone in the culture medium was the most favourable for the production of active fibrinolytic enzyme by Fusarium oxysporum N.R.C.1. Potassium dihydrogen phosphate had stimulating effect, while glucose, sucrose, lactose, ribose and soluble starch had adverse effect on enzyme productivity. The optimum of the fibrinolytic enzyme activity was at pH 7.0.

Purification and properties of two fibrinolytic enzymes from Fusarium oxysporum N.R.C.1.

The crude fibrinolytic enzyme preparation from Fusarium oxysporum N.R.C.1 was purified into two enzymes by ammonium sulphate precipitation followed by chromatography on Sephadex G-100 and DEAE-cellulose. Both fibrinolytic enzymes were more active on human than on bovine fibrin. The activity of the "major" enzyme component on human fibrin was 72-fold that of the "minor" enzyme component. Both enzymes had the same temperature (37 degrees C) and pH (6.98) optima. The "minor" enzyme component was more stable than the "major" one against heat and pH treatments. Both enzymes were significantly activated with Co2+ and inhibited with EDTA.

The biosynthesis of proteases with fibrinolytic activity in immobilized cultures of Penicillium chrysogenum H9.

The yield of fibrinolytic and other proteolytic enzymes by Ca-alginate immobilized cells of Penicillium chrysogenum H9 was compared to that of free cells. The gel matrix protected the immobilized cells from lysis providing higher stability to the biocatalyst. The highest fibrinolytic activity/caseinase activity ratio was obtained in immobilized cell cultures. The composition of culture medium influenced enzyme production, showing different patterns in free and immobilized cultures. An alginate concentration of 3% and 10 ml alginate beads/50 ml medium were optimum for fibrinolytic enzyme biosynthesis. Semicontinuous production of the enzyme indicated the superiority of immobilized cells. The beads were affected by repeated exposure to phosphate ions after 12 cycles.

Purification and some properties of pure Cochliobolus lunatus fibrinolytic Enzyme.

Purification of partially purified fibrinolytic enzyme was attempted by chromatography on DEAE-cellulose (D-52) column. The results indicated the resolution of three protein components and one minor component. It was shown that the first component was the major of the applied sample. Examination of fibrinolytic activity of the different fractions of components one and two indicated that only the first component possessed fibrinolytic activity. Fibrinolytic activity of the applied sample was completely recovered by the first enzyme component, and the most active fraction of this enzyme component showed 3.3-fold purification. The pure fibrinolytic enzyme was relatively more stable at pH 6.98, which was also optimal for its activity. After heating the enzyme solution (pH = 6.98) at 55 and 60 degrees C for 15 min, the enzyme still retained 34.7 and 17.3% of its original activity, respectively. Zinc ions partially inhibited the enzyme. Copper ions activated the enzyme. Iodine partially inhibited the fungal fibrinolytic enzyme at a final concentration of 10(-4)M; at 10(-2)M complete inactivation was brought about. The p-chloromercuribenzoate at a final concentration of 10(-2)M brought about partial inhibition whereby the enzyme lost about 33% of its original activity. Reduced glutathione brought about activation of the enzyme, while trypsin inhibitor did not show any effect on enzyme activity.

Preparation and properties of fibrinolytic enzymes produced by Cochliobolus lunatus.

Some properties of the crude lyophilized fibrinolytic enzyme produced by Cochliobolus lunatus in surface culture were studied. Enzyme concentrations over the range from 0.16 to 10.16 mg/mL showed that concentration above a certain level ceased to be the limiting factor controlling enzyme action. At pH 6.8 and a temperature of 40 degrees C, the fibrinolytic enzyme showed maximal activity at a human fibrin concentration of 2 mg/mL. The optimum pH values for enzyme activity were 6.98 and 7.0, using Sørensen and Mcllvaine buffers, respectively. Fibrinolytic enzymes were isolated from a static culture of Cochliobolus lunatus; isolation was carried out with various agents. Ammonium sulphate yielded the highest recovered fibrinolytic activity. The fraction salted out by precipitation at 25% ammonium sulphate saturation possessed the highest recovered fibrinolytic activity compared to the ammonium sulphate, ethanol, and acetone fractions.

Production and isolation of milk-clotting enzyme from Aspergillus versicolor.

The production of a milk-clotting enzyme by Aspergillus versicolor in 19 different culture media was investigated. Considerable milk-clotting activity was achieved by supplying corn steep liquor with either glucose of maltose. Dephytinization of corn steep liquor had an adverse effect on the production of milk-clotting enzyme. The results indicated that complex organic compounds favoured the production of the enzyme. Precipitation with acetone or tannin was unsuitable, but ammonium sulphate and ethanol above certain concentration produced active fractions.

Preparation and properties of pectic enzymes produced by Trichoderma lignorum.

Polygalacturonase and protein-methylesterase were isolated from shaken culture of Trichoderma lignorium. Isolation was carried out with various agents. Methanol was the most suitable precipitant for isolating polygalacturonase, yielding enzyme preparations 6.6 times more active than that of culture filtrate. Likewise, tannin afforded active fractions at pH 4 and 0.05% concentrations. Similarly, 50% ammonium sulphate saturation gave active fractions. The least polygalacturonase activity was obtained from ethanol. In any of the organic solvents used, highest enzymic activity was obtained when using only one volume. As regards pectin-methylesterase, no correlation existed between its activity and concentration of the precipitant used. A substrate concentration above 0.8% was a limiting factor for polygalacturonase activity, while optimum enzyme concentration was 40 microgram protein/ml at 40 degrees C and pH 4.45.

Effect of some agents on the activity of cell-free progesterone 11 alpha-hydroxylase and 11 beta-hydroxylase from Aspergillus niger 12Y.

The effect of some agents on the activity of cell-free progesterone 11 alpha-hydroxylase and 11 beta-hydroxylase from Aspergillus niger 12Y was studied. Calcium chloride, sodium chloride, magnesium sulphate, copper sulphate, and EDTA inhibited 11 alpha-hydroxylase and 11 beta-hydroxylase, while mercuric chloride inhibited only 11 alpha-hydroxylase. Inhibition of both the enzymes was also brought about by iodine, p-chloromercuribenzoate, iodoacetic acid, maleic acid, and cystine as well as potassium ferricyanide for 11 alpha-hydroxylase. Reduced glutathione and cysteine-HCl brought about activation of 11 alpha-hydroxylase and 11 beta-hydroxylase. The probability of the presence of reactive sulfhydryl groups in the active sites of both enzymes was discussed. Urea inhibited both fungal progesterone hydroxylases, probably due to enzyme protein denaturation.