Glutamine dipeptide attenuate mucosal atrophic changes and preservation of gut barrier function following 5-FU intervention

Traditional parenteral nutrition (PN) and chemotherapy may lead to changes mucosal morphology and gut barrier function. To investigate the effects of alanyl-glutamine on intestinal mucosal morphology and gut barrier function in PN-fed rats challenged with 5-FU male Wistar rats were central catheterized and randomily divided into two groups: PN group (n = 10) that received traditional parenteral nutrition solution only, and Ala-Gln group (n = 10) that received glutamine dipeptide enriched nutritional solution (3% Ala-Gln). The rats were maintained on their respective diets for 7 days. 5-FU (75 mg/kg) was injected intraperitoneally at 8 am on day 4. All rats were gavaged with lactulose (100 mg) and mannitol (50 mg) in 2ml before and three days after administration of 5-FU. Ala-Gln group maintained serum glutamine concentration, intestinal mucosal morphology. While bacterial translocation rates in ala-Gln group was 30%, in PN group was 90% (P < 0.05). Similar intestinal permeability was observed on day 3 in both groups. The control group had a significantly increased intestinal permeability on day 7 (P < 0.05), while Ala-Gln group maintained the intestinal permeability. It was suggested that alanyl-glutamine maintained intestinal morphology and gut barrier function in PN-fed rats challenged with 5-FU.     

The products of ribbon and raw are necessary for proper cell shape and cellular localization of nonmuscle myosin in Drosophila

Trehalose phosphorylase (EC 2.4.1.64) from Agaricus bisporus was purified for the first time from a fungus. This enzyme appears to play a key role in trehalose metabolism in A. bisporus since no trehalase or trehalose synthase activities could be detected in this fungus. Trehalose phosphorylase catalyzes the reversible reaction of degradation (phosphorolysis) and synthesis of trehalose. The native enzyme has a molecular weight of 240 kDa and consists of four identical 61-kDa subunits. The isoelectric point of the enzyme was pH 4.8. The optimum temperature for both enzyme reactions was 30 degrees C. The optimum pH ranges for trehalose degradation and synthesis were 6.0-7.5 and 6.0-7.0, respectively. Trehalose degradation was inhibited by ATP and trehalose analogs, whereas the synthetic activity was inhibited by P(i) (K(i)=2.0 mM). The enzyme was highly specific towards trehalose, P(i), glucose and alpha-glucose-1-phosphate. The stoichiometry of the reaction between trehalose, P(i), glucose and alpha-glucose-1-phosphate was 1:1:1:1 (molar ratio). The K(m) values were 61, 4.7, 24 and 6.3 mM for trehalose, P(i), glucose and alpha-glucose-1-phosphate, respectively. Under physiological conditions, A. bisporus trehalose phosphorylase probably performs both synthesis and degradation of trehalose.     

Clinical trials in the elderly. Should we do more?

In this study, SPF rat models were used. The purpose of the study was to observe the impairment of gut barrier function subsequent to long-term TPN, and evaluate the effect of TPN enriched by alanyl-glutamine (Ala-Gln) on the gut. After 7 day standard TPN, there was a significant decrease of CD3+, CD4+, CD8+ and IgA-containing plasma cells in lamina propria. The percentage of intestinal bacteria coated by S-IgA declined, and bacterial adherence to intestinal epithelial cells increased with an increased incidence of bacterial translocation to mesenteric lymph nodes. All these adverse effects could be attenuated by addition of Ala-Gln to TPN solutions or oral glutamine (Gln) or Ala-Gln administration. The results of the study suggested that long-term standard TPN impaired the immune gut barrier function and therefor facilitated enterogenic infection, and addition of Gln or Ala-Gln significantly benifited gut barrier protection and infection prevention, which might be important to clinical practice.     

Chemical degradation kinetics of recombinant hirudin (HV1) in aqueous solution: effect of pH

PURPOSE: To gain information on the chemical stability pattern and the kinetics of the degradation of recombinant hirudin variant HV1 (rHir), a thrombin-specific inhibitor protein of 65 amino acids, in aqueous solution as a function of pH. METHODS: Stability of rHir was monitored at 50 degrees C in the framework of a classical pH-stability study in aqueous buffers pH 1-9.5. Two capillary electrophoresis (CE) protocols were used: one for the kinetics of succinimide formation at Asp53-Gly54 (C-terminal tail) and Asp33-Gly34 (loop section), the other for the kinetics of rHir degradation. To check for potential effects of conformational changes by thermal denaturation, circular dichroism (CD) measurements were performed between 25 and 80 degrees C. RESULTS: Throughout the pH range studied no effect of thermal denaturation on rHir confirmation at 50 degrees C was observed. rHir was most stable at a neutral pH whereas, at slightly acidic pH, an intermediate stability plateau was found. Both, strongly acidic and alkaline conditions led to fast rHir degradation. Depending on the pH of degradation, rHir was found to degrade in various combinations of multiple parallel and sequential degradation patterns. Special focus was on succinimide formation at Asp53-Gly54 (C-terminal tail) and Asp33-Gly34 (loop) and on the potential of isoAsp formation in position 53 and 33. CONCLUSIONS: Chemical rHir stability in the intermediate pH range depends strongly on succinimide formation. At slightly acidic conditions succinimides represent the major degradation product (up to 40%). Around neutral pH succinimides react further, presumably by isoAsp formation, and concentrations remain low. Relative preference of succinimide formation in the C-terminal tail domain versus the loop domain is explained by higher backbone flexibility in the tail.     

Inactivation Kinetics of the Cyanobacterial Toxin Microcystin-LR by Free Chlorine

Worldwide, the increasing occurrence of toxins produced by cyanobacteria in water bodies used as source waters for drinking water has become an important public health issue. Microcystin-LR is one of the most commonly found cyanotoxins. A detailed evaluation of the free chlorine induced inactivation kinetics of extracellular microcystin-LR is presented in this study. Rate constants needed for chlorine inactivation of the toxin were derived from the data. The effects of varied pH, chlorine dose, toxin concentration, and temperature on the rate of inactivation were evaluated. Batch chlorination experiments were run using carbonate-buffered Milli-Q water at three different initial toxin concentrations (1, 2, and 8?μg/L), three different chlorine doses (1, 3, and 9?mg/L), and three different pH values (6.0, 7.5, and 9.0) at 11, 20 and 29°C. The study showed that extracellular microcystin-LR was inactivated by free chlorine and the inactivation rate was affected by pH. The highest inactivation rates were observed at pH 6.0 and the lowest at pH 9.0.     

Common mechanism of toxicity: a case study of organophosphorus pesticides

Despite the fact that glutamine is not considered to be an essential amino acid, it is the amino acid found in the greatest concentration both in plasma (26%) as in skeletal muscle (75%). These levels may decrease in post-operative, trauma, or critical patients. Glutamine performs many functions in which its demand may be increased, such as: it is a precursor of the synthesis of nucleotides; it is an activator of the protein synthesis and at the same time it inhibits the degradation; it is an activator of glycogen synthesis; it is a metabolic substrate for rapidly replicating cells; it is an energy source for the enterocyte which is so important for maintaining the integrity and the function of the intestinal barrier, and the consumption thereof may be increased under conditions of stress. The administration of glutamine intravenously leads to two physical-chemical problems; the first is its low solubility in water; at 20 degrees C this is only 36 g/l, and the second problem is its low chemical stability in an aqueous solution at 22-24 degrees C, this being 11 days. This problem has led the industry to research two dipeptides of glutamine; L-alanyl-glutamine, and L-glycyl L-glutamine, both of which are much more soluble and much more stable. At present there is still a controversy regarding the dosage of glutamine and its dipeptides, with the dose being 0.19-0.29 g/kg/day of L-glutamine or its dipeptide forms, in surgical post-operative periods or to prevent bacterial translocation, and in patients who are candidates for bone marrow transplants, the administered dose has been 0.37-0.57 g/kg/day. The purpose of this study is to review the existing bibliography regarding the efficacy of L-glutamine or its dipeptides in four possible indications for its application in the daily clinical practice, such as: a) In post-operative surgical patients of major or medium surgery, glutamine or its dipeptides reduces the losses of muscular glutamine and its catabolism, showing a less negative nitrogen balance. b) Whether it avoids bacterial translocation. c) Whether it favors the response of the immunological system. d) Whether in patients who are candidates for bone marrow transplants this decreases the side effects due to chemotherapy and radiotherapy such as mucositis, or whether it decreases the number of days of neutrophil recovery. At present, on the European market there are two commercially available brands of glutamine dipeptides: Dipeptiven, by Fresenius Laboratories, Germany. A 100 ml vial which corresponds to 20 g of L-alanyl L-glutamine (8.2 g of alanine + 13.46 g of L-glutamine). This is added to the standard amino acid solution. Glamin, Pharmacia and Upjohn Laboratory, Sweden. This is an amino acid solution with 13.4% essential and non-essential amino acids which are equivalent to 22.4 g of nitrogen/l, and which contain 30.27 g L-glycyl-L-glutamine (10.27 g of glycine + 20 g of L-glutamine).     

Chemical pathways of peptide degradation: IX. Metal-catalyzed oxidation of histidine in model peptides

PURPOSE: To elucidate the nature of the reactive oxygen species (i.e., superoxide anion radical, hydroxyl radical, and hydrogen peroxide) involved in the metal-catalyzed oxidation of histidine (His) in two model peptides. METHODS: The degradation of AcAla-His-ValNH2 (Ala-peptide) and AcCysNH2-S-S-AcCys-His-VaNH2 (Cys-peptide) was investigated at pH 5.3 and 7.4 in an ascorbate/cupric chloride/oxygen (ascorbate/ Cu(II)/O2) system, both in the absence and presence of selective scavengers (i.e., catalase, superoxide dismutase, mannitol, sodium formate, isopropanol, and thiourea) of the reactive oxygen species. All reactions were monitored by HPLC. The major degradation products were characterized by electrospray mass spectrometry. RESULTS: The Cys-peptide was more stable than the Ala-peptide at pH 5.3 and 7.4. Both peptides displayed greater stability at pH 5.3 than at 7.4. At pH 5.3, 35 +/- 0.7% of the Cys-peptide and 18 +/- 1% of the Ala-peptide remained after 7 hours, whereas at pH 7.4, 16 +/- 3% of the Cys-peptide and 4 +/- 1% of the Ala-peptide remained. Catalase, thiourea, bicinchoninic acid, and ethylenediaminetetraacetate were effective at stabilizing both peptides toward oxidation, while superoxide dismutase, mannitol, isopropanol, and sodium formate were ineffective. The main degradation products of the Ala- and Cys-peptides at pH 7.4 appeared to be AcAla-2-oxo-His-ValNH2 and AcCysNH2-S-S-AcCys-2-oxo-His-ValNH2, respectively. CONCLUSIONS: Hydrogen peroxide, Cu(I), and superoxide anion radical were deduced to be intermediates involved in the oxidation of the Ala- and Cys-peptides. Hydrogen peroxide degradation to secondary reactive oxygen species may have led to the oxidation of the peptides. The degradation of hydrogen peroxide by a Fenton-type reaction was speculated to form a complexed form of hydroxyl radical that reacts with the peptide before diffusion into the bulk solution.     

Effect of mucin and glucose on proteolytic and glycosidic activities of Streptococcus oralis

The production of glycosidase and protease activities, which may play a role in the degradation of human glycoproteins, by Streptococcus oralis strains isolated from endocarditis, septicaemia or the oral cavity was investigated with a range of fluorogenic substrates. The pH optima of the proteases ranged from 6.0 to 9.3 and the pH optima for the glycosidases were lower (4.5-6.0), although the pH range over which both groups of enzymes acted was broad. Growth in a minimal medium supplemented with glucose resulted in repression of glycosidase activities and elevated proteolytic activity. Bacteria from cultures supplemented with porcine gastric mucin (PGM), a model glycoprotein, exhibited higher levels of glycosidase activity, while proteolytic activity was suppressed and glycoprotein-derived monosaccharides were transported at significantly higher rates than those observed for cells grown in media with glucose. PGM-derived cells also exhibited high levels of N-acetylneuraminate pyruvate-lyase, the first intracellular enzyme in the pathway of sialic acid catabolism. Taken together, these data indicate that S. oralis strains produce a range of proteolytic and glycosidic enzymes that may play a role in the degradation of host-derived glycoproteins.     

Glutamine dipeptides in clinical nutrition

Glutamine is a conditional indispensable amino acid during stress. However, limited solubility and instability of glutamine prevent its addition to presently available nutritional preparations. To overcome these drawbacks, we propose the dipeptide concept by which stable and highly soluble synthetic glutamine containing dipeptides are used. The synthetic dipeptides fulfill all chemical/physical properties to be considered as parenteral substrates. Numerous experimental studies show rapid clearance of parenteral supplied glutamine containing dipeptides without accumulation in tissues; the loss via the urine being inconsequential. Differences related to the dipeptide structure are not observed. There is overwhelming evidence existent that a nutritional support with supplemental glutamine dipeptide positively influences nitrogen excretion, immune status, gut integrity, morbidity, rehabilitation and outcome. Consequently, omission of glutamine from conventional TPN and its subsequent administration should be considered as a replacement of a deficiency rather than a supplementation. It might thus be conceivable that the beneficial effects observed with glutamine nutrition are simply a correction of disadvantages produced by an inadequacy of conventional amino acid solutions. The availability of stable glutamine containing preparations will certainly facilitate an adequate amino acid nutrition in routine clinical setting during episodes of stress and malnutrition.     

pH dependence of facilitation by neurotransmitters and divalent cations of P2X2 purinoceptor/channels

The pH dependence of the facilitation by dopamine (10 microM), 5-hydroxytryptamine (10 microM), adenosine (1 and 100 microM), Zn2+ (10 microM) and Cd2+ (1 mM) of P2X2 purinoceptor/channels was tested by expressing these channels in Xenopus oocytes. In a pH range between 6.0 and 8.5, concentration-response curves for an inward current activated by ATP were shifted toward a lower concentration range at a more acidic pH, indicating that the sensitivity to ATP is pH-dependent. Comparison of the effects of the neurotransmitters and the divalent cations on the ATP-activated current was made using a concentration of ATP which activated 40-50% of the maximal current at each pH value. The current facilitation by dopamine was obvious at pH 7.1 and 7.7, but was not observed at pH 8.5. At pH 6.0, the current was inhibited upon first trials of dopamine, but it was facilitated upon second trials. With 5-hydroxytryptamine and adenosine, the current facilitation was most remarkable at pH 6.0, less remarkable at pH 7.1 and 7.7, and the facilitation was almost abolished at pH 8.5. On the other hand, the current facilitation by Zn2+ and Cd2+ was more remarkable at alkaline pH values (7.7 and 8.5), and the facilitation was almost abolished at pH 6.0. The results suggest that the facilitation of P2X2 purinoceptors depends on pH, and the pH dependence was different between the neurotransmitters and the divalent cations.     

Enantioseparation of amino acids and dipeptides using vancomycin as chiral selector in capillary electrophoresis

Vancomycin was applied as chiral selector for the enantiomeric separation of derivatized amino acids and dipeptides. The influence of vancomycin concentration, pH and presence of 2-propanol in the buffer were examined in order to find optimal separation conditions. Optimization was by factorial design. Further, chiral separation of derivatives prepared with three different reagents was compared. These reagents were 9-fluorenylmethyl chloroformate (FMOC), 2-(9-anthryl)ethyl chloroformate (AEOC) and dansyl chloride (dansyl). Optimum resolution was at high vancomycin concentrations, while optimum efficiency was at low vancomycin concentrations. As a consequence of the very high enantioselectivity of vancomycin, the vancomycin concentration below the amount necessary for maximal resolution can be used. Separation efficiency was relatively low, and this could be attributed to adsorption of the selector at the capillary wall. Three factors led to decreased adsorption: application of a pH above the zero mobility pH value, low vancomycin concentrations and the presence of 2-propanol. For amino acids, the resolutions of the different derivatives were: dansyl > AEOC > FMOC, while for dipeptides, the highest selectivity was with AEOC.     

Degradation of MTBE Intermediates using Fenton’s Reagent

In a previous study, the chemical oxidation of methyl tert-butyl ether (MTBE) at low concentrations in water using Fenton’s reagent (FR) was investigated. At certain reaction conditions the process achieved 99.99% degradation of MTBE but it did not result in complete MTBE mineralization. In the present study, the major intermediate by-products generated during the reaction, such as tert-butyl formate (TBF), tert-butyl alcohol (TBA), methyl acetate, and acetone were separately used as parent contaminants and treated under the same reaction conditions initially used for MTBE (i.e., pH of the water, molar ratio of pollutant to FR) in order to compare their degradability by hydroxyl radicals generated from Fenton’s reaction. The results were compatible with the second order reaction rate constants for the reaction of hydroxyl radicals with each contaminant commonly available in the literature. The comparison of the degradation kinetics for each intermediate by-product provided information that aims at unveiling the limiting step(s) of the entire MTBE degradation pathway. In this context, it was found that (1) TBA was generated by reactions subsequent to those that produced TBF, (2) acetone was originated by at least three independent pathways involving direct hydroxyl radical attack on MTBE, TBF, and TBA, and (3) methyl acetate was formed exclusively from MTBE.     

Preflight versus en route success and complications of rapid sequence intubation in an air medical service

Methacholine chloride bronchoprovocation challenges are performed for the diagnosis and investigation of hyperreactive airways. Over the last 20 yrs various formulations and pH values for the preparation of solutions of methacholine have been described. To determine the stability of methacholine chloride solutions prepared in a variety of buffers with differing pH values and under varying storage temperatures, we measured methacholine concentrations at intervals from 1 to 5 weeks. It was found that methacholine chloride solutions rapidly decompose if the pH is greater than 6 and that decomposition is more rapid as the pH is raised; solutions at pH 9, i.e. bicarbonate buffer, and stored at 27 degrees C have degradation up to 36% after only one week. Solutions of the same pH but prepared in different buffers can have both varied rates of deterioration and different absolute amounts of methacholine hydrolysed, e.g. solutions prepared in pH 9 borate buffer and stored at 27 degrees C have up to 60% degradation after 1 week. Solutions prepared in saline are stable probably because methacholine solutions are weakly acidic. The results emphasise the importance of preparing methacholine chloride in the proper buffers for use in the accurate assessment of airway responsiveness.     

Prognostic factors and rational approach in the treatment of submucosal cancer of the stomach

Metallothionein (MT) mRNA levels were analyzed following exposure of neonatal rat primary astrocyte cultures to physiologic pH (7.4), acidosis (pH 6.5 and 6.0), and dimethyl sulfoxide (DMSO). Treatments were carried out both in the presence and absence of the bioflavonoid, quercetin. Total RNA was probed on northern blots with [alpha32P]dCTP-labeled synthetic cDNA probes specific for rat MT isoform mRNAs. MT-I and MT-II mRNA levels in astrocytes exposed to pH 6.5 or pH 6.0 were increased compared to controls (pH 7.4). Treatment with DMSO in the presence and absence of acidosis, also increased MT-I and MT-II mRNA levels compared to controls (pH 7.4). The DMSO-induced increase in MT mRNA expression was reversed by treatment of astrocytes with quercetin, such that MT-I and MT-II mRNA levels in DMSO plus quercetin-treated astrocytes were indistinguishable from mRNA levels in their respective controls at pH 7.4, pH 6.5, and pH 6.0. These findings suggest that both acidosis and DMSO exposure are associated with increased astrocytic MT synthesis at the mRNA level, and that quercetin, effectively blocks MT mRNA induction by DMSO.     

Effects of methionine sulphoximine treatment on renal amino acid and ammonia metabolism in rats

Renal glutamine metabolism in relation to ammoniagenesis has been extensively studied during chronic metabolic acidosis, when arterial glutamine levels are reduced. However, little is known about the effects of reduced glutamine delivery on renal glutamine and ammonia metabolism at physiological systemic pH values. Therefore, a model of decreased arterial glutamine concentrations at normal pH values was developed using methionine sulphoximine (MSO). Renal glutamine and ammonia metabolism was measured by determining fluxes and intracellular concentrations after an overnight fast in ether anaesthetized normal rats, MSO-treated rats and their pair-fed controls. Moreover, fluxes and intracellular concentrations of several other amino acids were determined concomitantly. After 2 and 4 days of MSO treatment, arterial glutamine concentrations were reduced to 55%, while arterial ammonia concentrations increased by 70%. Kidney glutamine uptake reduced, but systemic pH was unchanged. Fractional extraction of glutamine remained unchanged, suggesting that also in vivo net uptake of glutamine by the kidney at subnormal levels is related to arterial glutamine concentrations. As a result, at day 2 but not at day 4, the kidney reduced the net release of ammonia into the renal vein and thus reduced net renal ammonia addition to body ammonia pools. Therefore at day 2, the kidney seems to play an important role in adaptation to both hyperammonaemia and hypoglutaminaemia.     

Stability of cephalosporin prodrug esters in human intestinal juice: implications for oral bioavailability

The levels of degradation of cefetamet pivoxil (CAT), cefuroxime axetil (CAE), and cefpodoxime proxetil (CPD) in 0.6 M phosphate buffer (pH 7.4) and human intestinal juice (pH 7.4) at 37 degreesC over 24 h were compared. Significant differences in the time courses of degradation and in the patterns of degradation products were observed. (i) The relative proportions of the Delta2- and Delta3-cephalosporins were roughly reversed in the two incubation media. In phosphate buffer, the major degradation product was the Delta2-cephalosporin (CAT = 61%; CAE = 74%; CPD = 85%), while in intestinal juice it was the Delta3-cephalosporin (CAT = 86%; CAE = 75%; CPD = 87%). (ii) Generally, the degradation of the prodrug esters progressed faster in intestinal juice than in phosphate buffer (e.g., for CAT the half-lives [t1/2s] were 0.78 and 4.3 h, respectively). (iii) The two diastereoisomers of CAE and CPD were degraded at different rates in intestinal juice (for the CAE diasteroisomers, t1/2s = 0.37 and 0.93 h; for the CPD diastereoisomers, t1/2s = 0.18 and 0.98 h) but were degraded at similar rates in phosphate buffer (for the CAE diastereoisomers, t1/2 = 1.6 h; for the CPD t1/2 diastereoisomers, = 2.2 h). It is concluded that (i) the Delta2 isomerization does not significantly affect the bioavailability of prodrug esters since enzymatic hydrolysis in the intestinal fluid proceeds mainly to the active Delta3-cephalosporin and (ii) the high degree of stereoselectivity of the enzymatic ester hydrolysis should make it possible to increase the bioavailabilities of certain prodrug esters (CAE, CPD) by using the more stable diasterioisomer.     

Alanine and glutamine kinetics at rest and during exercise in humans

PURPOSE: The purpose of this study was to quantify both alanine and glutamine kinetics during exercise of moderate intensity to determine the sum total of alanine and glutamine flux. METHODS: Tracer methods were used to quantify alanine and glutamine rates of appearance (Ra) in plasma at rest and during 180 min of approximately 45% VO2max treadmill exercise in six normal volunteers (25 +/- 2 yr, 68 +/- 2.5 kg, VO2max 43 +/- 2.4 mL.min-1.kg-1; means +/- SE). Bolus injections (N = 3) or primed-constant infusions (N = 3) of 2H5-glutamine and 3-13C-alanine were given at rest on 1 d and 10-15 min after the onset of exercise on a separate day less than 2 wk later. Plasma enrichment decay curves and plateau enrichments were used to estimate alanine and glutamine kinetics. RESULTS: Whereas alanine Ra increased significantly from rest to exercise (5.72 +/- 0.31 vs 13.5 +/- 1.9 mumol.min-1.kg-1, respectively; P < 0.01), glutamine Ra was not significantly altered by exercise (6.11 +/- 0.44 and 6.40 +/- 0.69 mumol.min-1.kg-1 at rest and during exercise, respectively). The total of alanine and glutamine flux increased from 17.93 +/- 0.88 to 25.98 +/- 3.04 (P < 0.05). CONCLUSIONS: Since most muscle amino-N is released as alanine and glutamine, these findings provide strong evidence that amino-N delivery from muscle to the liver is increased during exercise. In addition, it appears that alanine, rather than glutamine, is the predominant N carrier involved in the transfer of N from muscle to the liver during moderate intensity exercise.     

Effects of GABAB receptor antagonism on the development of pentylenetetrazol-induced kindling in mice

In our previous studies, the yeast Endomyces fibuliger LU677 was found to degrade amygdalin in bitter apricot seeds. The present investigation shows that E. fibuliger LU677 produces extracellular beta-glycosidase activity when grown in malt extract broth (MEB). Growth was very good at 25 degrees C and 30 degrees C and slightly less at 35 degrees C. When grown in MEB of pH 5 and pH 6 with addition of 0, 10 or 100 ppm amygdalin, E. fibuliger produced only slightly more biomass at pH 5, and was only slightly inhibited in the presence of amygdalin. Approximately, 60% of the added amygdalin was degraded (fastest at 35 degrees C) during an incubation period of 5 days. Supernatants of cultures grown at 25 degrees C and pH 6 for 5 days were tested for the effects of pH and temperature on activity (using amygdalin, linamarin and prunasin as substrates). Prunase activity had two pH optima (pH 4 and pH 6), amygdalase and linamarase only one each at pH 6 and pH 4-5 respectively. The linamarase activity evolved earlier than amygdalase (2 days and 4 days respectively). The data thus indicate the presence of at least two different glycosidases having different pH optima and kinetics of excretion. In the presence of amygdalin, lower glycosidase activities were generally produced. However, the amygdalin was degraded from the start of the growth, strongly indicating an uptake of amygdalin by the cells. The temperature optimum for all activities was at 40 degrees C. Activities of amygdalase (assayed at pH 4) and linamarase (at pH 6) evolving during the growth of E. fibuliger were generally higher in cultures grown at 25 degrees C and 30 degrees C. TLC analysis of amygdalin degradation products show a two-stage sequential mechanism as follows: (1) amygdalin to prunasin and (2) prunasin to cyanohydrin.     

Kinetics and products of the TiO2 photocatalytic degradation of 2-chlorobiphenyl in water

The light-induced degradation of 2-chlorobiphenyl (2-CB) under simulated solar irradiation has been investigated in aqueous solutions containing TiO2 suspensions as photocatalysts. The apparent quantum yield for an initial 2-CB concentration C0 = 3.8 micrograms/mL at the natural pH was ca. 0.005 The oxidation kinetics of 2-CB follows the Langmuir-Hinshelwood kinetic model at natural pH. The primary degradation of 2-CB follows a pseudo-first-order kinetics. Several reaction intermediates were identified using GC/FTIR/MS and ion chromatography. The products at the initial stage of the reaction were seven isomers of 2-chlorobiphenyl-ol and biphenyl-2-ol. These intermediates underwent further photocatalytic oxidation via aldehydes, ketones, and acids finally into CO2 and HCl. The formation and fate of some of these compounds under irradiation were also investigated. A reaction scheme involving hydroxyl radicals has been proposed.