Optimization of cultural conditions of Arthrobacter sp. Sphe3 for growth-associated chromate(VI) reduction in free and immobilized cell systems.

The current study aimed to characterize Arthrobacter sp. Sphe3 ability to reduce Cr(VI) in suspended cell cultures as well as in immobilized form using Ca-alginate beads. Adaptation studies in the presence of 5 mg L(-1) Cr(VI) showed a significant increase in specific growth rate from 0.25 to 0.3 h(-1) and bioremoval percentage from 64% to 94% (p<0.05), whereas Arthrobacter sp. Sphe3 could tolerate up to 50 mg L(-1) Cr(VI). Optimization of culture conditions resulted in complete reduction of 45 mg L(-1) Cr(VI) at 30 °C, pH 8 and 10 g L(-1) of glucose. High glucose concentrations helped at reducing (80±2.4)% of initial 100 mg L(-1) Cr(VI), whereas the bacterial strain could tolerate 850 mg L(-1) Cr(VI). Cr(III) formation was first evidenced by the appearance of a green insoluble precipitate in the medium. Cell biomass was successfully immobilized in Ca-alginate beads that were evaluated for their stability. Cell release was sharply decreased when 4% Na-alginate was used under non-shaking conditions. Biotransformation efficiency was enhanced when 25-50 mg cells mL(-1) Na-alginate from the exponential growth phase were collected and co-encapsulated with either 1% glucose and 0.5% (NH4)2SO4, or 1% LB medium. Immobilized biocatalyst could be reused up to 6 continuous cycles in the presence of 10 mg L(-1) Cr(VI), but its performance was lowered at higher metal concentrations comparing with free cells that significantly maintained their reducing ability up to 300 mg L(-1) Cr(VI).

Equilibrium, thermodynamic and kinetic studies on biosorption of Mn(II) from aqueous solution by Pseudomonas sp., Staphylococcus xylosus and Blakeslea trispora cells.

Biosorption of Mn(II) from aqueous solutions using Pseudomonas sp., Staphylococcus xylosus and Blakeslea trispora cells was investigated under various experimental conditions of pH, biomass concentration, contact time and temperature. The optimum pH value was determined to 6.0 and the optimum biomass concentration to 1.0 g L(-1) for all types of cells. Mn(II) biosorption was found to fit better to the Langmuir model for Pseudomonas sp. and B. trispora and to Freundlich model for S. xylosus. Langmuir model gave maximum Mn(II) uptake capacity 109 mg g(-1) for Pseudomonas sp. and much lower, 59 mg g(-1) and 40 mg g(-1) for S. xylosus and B. trispora, respectively. Pseudo-second-order kinetic model was also found to be in good agreement with the experimental results. Thermodynamic parameters of the adsorption confirmed the endothermic nature of sorption process with positive heat of enthalpy, accompanied by a positive value of entropy change. Interestingly, desorption experiments by treating biomass with 0.1 M HNO(3) solution resulted to more than 88% recovery of the adsorbed Mn(II) from Pseudomonas sp. and almost 95% and 99% from S. xylosus and B. trispora cells respectively, thus indicating that Mn(II) can be easily and quantitatively recovered from biomass.

Co-metabolism of 2,4-dichlorophenol and 4-Cl-m-cresol in the presence of glucose as an easily assimilated carbon source by Staphylococcus xylosus.

Comparison of the ability of Staphylococcus xylosus to degrade 2,4-dichlorophenol and 4-Cl-m-cresol in separate cultures is reported. Bacterial adaptation and the continuous presence of glucose, as a conventional carbon source, were found to stimulate the degrading efficiency of S. xylosus. 4-Cl-m-cresol exhibited higher substrate-induced toxicity with K(ig) value at 0.25 mM, comparing to 2,4-dichlorophenol (K(ig) value at 0.90 mM) at initial concentration ranging from 0.1 to 0.5 mM. Degradation rate of 4-Cl-m-cresol was found to decrease only, revealing lower value of inhibition degradation constant (K(i) at 0.019 mM) comparing to that of 2,4-dichlorophenol (K(i) at 0.41 mM). Both glucose and each one of the chloro-aromatic compounds tested were simultaneously consumed and an increase of chloride ions in the medium appeared, during the exponential phase of growth. The chloride ions increase was nearly stoichiometric in the presence of 2,4-dichlorophenol and one of its several intermediate products identified was 2-Cl-maleylacetic acid. In the case of 4-Cl-m-cresol, only one metabolic product was found and identified as 3-methyl-4-oxo-pentanoic acid.

Effect of biomass pre-treatment and solvent extraction on beta-carotene and lycopene recovery from Blakeslea trispora cells.

The production of carotenoids from Blakeslea trispora cells in a synthetic medium has been reported, with the main products being beta-carotene, lycopene, and gamma-carotene. The effect of biomass pretreatment and solvent extraction on their selective recovery is reported here. Eight solvents of class II and III of the International Conference of Harmonization: ethanol, methanol, acetone, 2-propanol, pentane, hexane, ethyl acetate, and ethyl ether, and HPLC analysis were used for the evaluation of their selectivities towards the three main carotenoids with regard to different biomass pre-treatment. The average C(max) values (maximum concentration of caronoids in a specific solvent) were estimated to 16 mg/L with the five out of eight solvents investigated, whereas methanol, pentane, and hexane gave lower values of 10, 11, and 9 mg/L, respectively. The highest carotenoid yield was obtained in the case of wet biomass, where 44-56% is recovered with one solvent and three extractions and the rest is recovered only after subsequent treatment with acetone; thus, four extractions of 2.5 h are needed. Two extractions of 54 min are enough to recover carotenoids from dehydrated biomass, with the disadvantage of a high degree of degradation. Our results showed that, for maximum carotenoid recovery, ethyl ether, 2-propanol, and ethanol could be successfully used with biomass without prior treatment, whereas fractions enriched in beta-carotene or lycopene can be obtained by extraction with the proper solvent, thus avoiding degradation due to time-consuming processes.

Kinetics of 2,4-dichlorophenol and 4-Cl-m-cresol degradation by Pseudomonas sp. cultures in the presence of glucose.

Comparison of the ability of Pseudomonas sp. to degrade 2,4-dichlorophenol and 4-Cl-m-cresol in separate cultures in the presence of glucose, as a conventional carbon source, is reported. The specific growth rates at 0.1 mM 2,4-dichlorophenol and 4-Cl-m-cresol were estimated to be 0.181 and 0.154 h(-1), respectively, showing that Pseudomonas sp. is mainly inhibited by 4-Cl-m-cresol. The percentage of consumption ranges between 65% and 11% for 2,4-dichlorophenol and between 37% and 8% for 4-Cl-m-cresol, respectively, depending on its initial concentration. The dechlorination of the two compounds was investigated in the growth media and it was found that chloride liberation in the case of 2,4-dichlorophenol took place during the exponential phase of growth, followed by pH decrease from 6.1 to 5.8 at 0.1 mM. In contrast, in the case of 4-Cl-m-cresol chloride ion release was observed to a lesser extent, indicating the different metabolic pathway of 4-Cl-m-cresol. 2,4-Dichlorophenol and 4-Cl-m-cresol degradation followed a first-order kinetics model, whereas glucose consumption fitted well a zero-order kinetics model.

Selected eHealth applications in Cyprus from the training perspective.

OBJECTIVES: In this paper a review of selected eHealth applications in Cyprus is presented linked with their success or failure based on their training activities. METHODS: The eHealth systems presented and their training activities include an update of the health information system (HIS) in the public hospitals, a medical system for emergency telemedicine (AMBULANCE and EMERGENCY-112 projects), a home monitoring system for cancer patients (DITIS), a satellite-based network in healthcare applications (EMISPHER and HEALTHWARE projects), and the training activities of the Cyprus Society of Medical Informatics. Different methodologies for training were used ranging from classical approaches like train the trainers, using demo cases followed by personal training, group training, and workshops, to more recent methodologies based on eLearning sessions including teleconsultations. RESULTS: The training was carried out successfully in all cases. However, not all eHealth systems were put into practice successfully, mainly for reasons not related to training. CONCLUSIONS: It is anticipated that this paper will promote the importance of these applications and their training activities as well as help in the spin off of others thus enabling the offering of a better service to the citizen.

Comparative study of Cd(II) and Cr(VI) biosorption on Staphylococcus xylosus and Pseudomonas sp. in single and binary mixtures.

Biosorption of Cd(II) and Cr(VI) ions in single solutions using Staphylococcus xylosus and Pseudomonas sp., and their selectivity in binary mixtures was investigated. Langmuir and Freundlich models were applied to describe metal biosorption and the influence of pH, biomass concentration and contact time was determined. Maximum uptake capacity of cadmium was estimated to 250 and 278 mg g(-1), whereas that of chromium to 143 and 95 mg g(-1) for S. xylosus and Pseudomonas sp., respectively. In binary mixtures with Cd(II) ions as the dominant species, there is a profound selectivity for cadmium biosorption, reaching 96% and 89% for Pseudomonas sp. and S. xylosus, respectively, at 10 mg l(-1) Cd(II) and 5 mg l(-1) Cr(VI). Interesting, when chromium (VI) ions are the dominant species, there is selectivity towards chromium around 92% with S. xylosus only.

Conjugation of resveratrol with RGD and KGD derivatives.

The reaction between Arg-Gly-Asp (RGD) and Lys-Gly-Asp (KGD) derivatives with 3,4',5-trihydroxy-trans-stilbene (resveratrol) was investigated. Knowing that resveratrol, RGD as well as KGD analogues inhibit human platelet aggregation in vitro, it was tempting for us to examine whether their coupling products present enhanced biological activity. Here, we report on the synthesis and identification of these coupling products. The N-protected peptides were synthesized by solid phase technique, using the 2-chlorotrityl-chloride resin, by the method of carbodiimides. Coupling reactions with resveratrol took place in solution using N,N-dicyclohexylcarbodiimide (DCC) as coupling reagent and 4-dimethylaminopyridine (DMAP) as catalyst. The reaction products were purified by reversed phase HPLC and identified by ESI-MS. The mono-esterified resveratrol derivative was the main (or only) reaction product, whereas the di- and the tri-ester (to a less extent) formation was noticed in some cases.

Synthesis and cytogenetic effects of aminoquinone derivatives with a di- and a tripeptide.

Quinones are of significant interest due to their important role in specific cellular functions. Quinoproteins are a big class of oxyreductive agents occurring in bacteria and other organisms. In this investigation derivatives of 2-amino-1,4-benzoquinone, 2-amino-1,4-naphthoquinone and 2-amino-5,8-dihydroxy-1,4-naphthoquinone with a di- and a tripeptide were prepared for first time. The effect of the synthesized compounds on sister chomatid exchange (SCE) rates and human lymphocyte proliferation kinetics on a molar basis was studied. Among these coupled products the most effective in inducing SCEs and depressing proliferation rate indices is the coupling product of 2-amino-1,4-naphthoquinone with the tripeptide GHK (10). Next in order of magnitude in inducing cytogenetic effects is 2-amino-1,4-naphthoquinone (2) and its coupling products with glycine and serine (4 and 5), while the rest displayed marginal activity.

Volume and secretion rate of lung liquid in the final days of gestation and labour in the fetal sheep.

1. Most of the liquid that fills the lung of the fetal sheep in late gestation is cleared by the end of labour. Clearance of this liquid has a beneficial effect on postnatal gas exchange and therefore represents an important adaptation for postnatal life. Despite its importance, there is disagreement about whether clearance begins prior to labour, or occurs entirely within labour. 2. To address this issue, we made serial determinations of lung liquid volume by indicator dilution during late gestation and labour in the fetal sheep. 3. Regression analysis demonstrated that lung liquid volume exhibited a plateau level in the near-term fetus before it began to decline. Two models provided a fit to the decline in volume. In one, lung liquid clearance occurred in two linear phases, the first beginning 70 h before the study was terminated when the ewe was in advanced labour, the second occupying the last 8 h of the study period. In the initial phase, average lung liquid volume fell from 38.3 to 26.4 ml x kg(-1) before a rapid decline in the second phase reduced the volume to 13.8 ml x kg(-1). An exponential decay model was also found to fit the data; this showed a gradual decline in lung liquid volume in the 2 days preceding onset of labour, followed by a much more rapid decline within labour. 4. The rate of lung liquid secretion also declined in two linear phases, both of which commenced earlier than the changes in lung liquid volume. An exponential decay model also gave a significant fit to the data, but the fit was significantly weaker than that achieved with the two-slope model. 5. We conclude that clearance of lung liquid begins well before commencement of labour in the full term fetal sheep, and then accelerates once labour is established. In our study, lung liquid volume fell even in the absence of reabsorption of liquid across the pulmonary epithelium, indicating that outflow of liquid through the trachea must have occurred at a rate in excess of the secretion rate.

Influence of prenatal adrenaline infusion on arterial blood gases after caesarean delivery in the lamb.

The efficacy of pulmonary gas exchange immediately after delivery is inversely related to the volume of liquid in the lung at birth, but aspiration of as much liquid as possible from the lung before Caesarean delivery fails to improve postnatal oxygenation (Pa,O2) to the level achieved after spontaneous term delivery. We hypothesised that the differing respiratory benefit of aspiration and vaginal delivery results from the differing volume of lung liquid remaining after aspiration (17 ml (kg body weight)-1) and labour (7 ml kg-1). We addressed this hypothesis by reducing lung liquid volume to an estimated 7 ml kg-1 by infusing adrenaline to seven fetal lambs at 140 days gestation (term is 147 days) before performing Caesarean delivery and obtaining postnatal blood gases for comparison with samples from lambs delivered vaginally. Infusion of adrenaline to fetuses caused a progressive decline in arterial O2 saturation (Sa, O2), pH and base excess, but no change in arterial partial pressure of O2 (Pa,O2) or CO2 (Pa,CO2). After birth, Pa,O2 rapidly rose to the same level in adrenaline-treated and vaginal-delivery groups. A severe acidosis occurred in the adrenaline-treated group and this appeared to be related to a higher Pa,CO2 and a transiently lower Sa, O2 in this group. We conclude that adrenaline infusion can enhance postnatal Pa,O2 levels in the newborn lamb, but this beneficial effect may be outweighed by the severe acidosis that develops after prolonged prenatal adrenaline treatment.

Errors in estimating lung liquid volume in fetal lambs when using radiolabeled serum albumin and blue dextran.

Fetal lung liquid volume is usually determined by using radio-iodinated serum albumin (RISA) or blue dextran (BD) as volume tracers. We tested the reliability of both tracers at 124 (G124) and 142 days of gestation (G142; term = G147) when the labels were employed simultaneously. We measured the proportion of label bound reversibly to the lung, or apparently lost from the lung compartment, by washing out the lung with saline and 5% albumin. At G124, volume estimates with the two labels were similar. At G142, the volume estimate with BD (36.3 +/- 8.7 ml/kg of body wt) was higher (P < 0. 05) than with RISA (22.3 +/- 3.5 ml/kg). This difference resulted from reversible binding of BD, because 5% albumin washout released 38.5 +/- 4.0% of the BD added at the start of the experiment but a lesser amount of RISA (9.8 +/- 0.7%; P < 0.05). At G142, when RISA was used alone, its reversible binding was 1.3 +/- 0.2%. Background absorbance increased during experiments, giving rise to an apparent increase in BD concentration. We conclude that RISA is an effective tracer for lung liquid volume determination in the fetal lamb, whereas our findings of substantial epithelial binding of BD and large changes in background absorbance demonstrate that, under the conditions of our experiments, BD is a poor tracer close to term.

Spectral analysis of a series of partially protected and deprotected tetrapeptides, analogues of AS-I phytotoxin.

A series of six tetrapeptides, analogues of AS-I phytotoxin, pathogenic to sunflower, have been synthesized either in solution and/or by solid phase methods and have been tested for phytotoxic activity in various plants and cytotoxic activity in three cancer cell lines. These peptides were identified as model compounds by fast atom bombardment (FAB), plasma desorption (PD), electrospray ionization (ESI) mass spectrometry and by 1H, 1H-1H, 13C and 1H-13C NMR. The data presented show that in protected tetrapeptides the molecular ion was easily identified whereas some difficulties appeared with the fully deprotected peptides. NMR spectra are given.

Cu(III)-polypeptide complexes exhibiting SOD-like activity.

The SOD-like activity of Cu(III)-complexes with polypeptides poly-L-lysine and poly-L-glutamic acid respectively was investigated. The Cu(II)-polypeptide complexes were first oxidized by K2IrCl6 to give the corresponding Cu(III)-compounds. The oxidation of Cu(II) and the corresponding Cu(II)/Cu(III) potential was evaluated by cyclic voltammetry (c.v.), UV-Vis and EPR spectroscopic (r.t.) experiments. Spin trapping EPR spectra were also conducted to confirm the formation of the superoxide radical. The SOD-like activity of each Cu(III)-complex was proved using the nitro blue tetrazolium (NBT) method slightly modified.

Xanthan gum production by Xanthomonas campestris w.t. fermentation from chestnut extract.

Xanthomonas campestris w.t. was used for production of xanthan gum in fermentations with chestnut flour for the first time. Fermentations were carried out with either chestnut flour or its soluble sugars (33.5%) and starch (53.6%), respectively, at 28 degrees C and 200 rpm at initial pH 7.0 in flasks. The effect of agitation rate (at 200, 400, and 600 rpm) on xanthan gum production was also studied in a 2-L batch reactor. It was found that xanthan production reaches a maximum value of 3.3 g/100 mL at 600 rpm and 28 degrees C at 45 h.

Antiproliferative activity of synthetic tetrapeptides, analogs of AS-I phytotoxin, towards cancer cell lines.

The in vitro chemosensitivity of three cancer cell lines [HT29 (colon), HeLa (cervical) and T47D (breast)] to eight synthetic tetrapeptides, analogs of AS-I toxin, with phytotoxic effect on a series of plants was studied. Mouse fibroblast L929 cell line was also tested for chemosensitivity to these peptides. All cell lines were especially sensitive to Cys-Val-Gly-Glu tetrapeptide with IC50 values of 0.18, 0.3 and 0.63 mM for HT29, HeLa and T47D cells, respectively, whereas the IC50 value for the L929 cells was higher than 1 mM. Antiproliferative activity was also observed with peptides Tyr-Val-Gly-Glu and His-Val-Gly-Glu with IC50 values higher than those obtained for Cys-Val-Gly-Glu. For the rest of the peptides tested the IC50 values were found close to or higher than 3 mM.

Massive decline in lung liquid before vaginal delivery at term in the fetal lamb.

OBJECTIVE: Our aim was to determine the volume of liquid remaining in the lungs of the fetal lamb just before a normal vaginal delivery at term to assess the extent to which an excess of liquid in the airspaces might contribute to the respiratory morbidity that accompanies elective cesarean delivery. STUDY DESIGN: The volume of liquid in the future airspace of the lungs was determined at the end of labor in eight fetal lambs at term from the dilution of an impermeable tracer (125I-labeled human serum albumin) mixed into the liquid. This volume was compared with that measured in a second group of 10 fetal lambs studied 7 days before the expected date of delivery (term = 147 days). RESULTS: The volume of lung liquid present at the end of labor was 6.8 +/- 1.0 ml x kg(-1) (n = 8) compared with 28.2 +/- 1.8 ml x kg(-1) (n = 10) in the second group of lambs studied before the onset of labor at 140 days of gestation. CONCLUSION: Our results indicate that the bulk (>75%) of the liquid that fills the lungs of the fetal lamb at 140 days of gestation is cleared at some time before normal term birth, suggesting that the adverse respiratory impact of elective cesarean delivery may be largely explained by denying the fetus this important adaptive mechanism.