Saline stress and cell toxicity evaluation using suspended plant cell cultures of horticultural crops grown in a bioreactor

Crop salt damage consists, usually, of leaf burn and defoliation, and it is associated with accumulation of toxic levels of sodium and/or chloride in leaf cells (Storey and Walker, 1999). The cell and tissue culture are simple biological systems that offer a direct approach to the metabolic changes. The plant cell growth in a controlled environment, as a bioreactor, is a unique tool for cell ion transport studies. Cell suspension culture of citrus cell line was exposed to a medium containing different sodium chloride concentrations (0mM, 42.7mM and 85.5mM). The growth profile of control cells (absence of NaCl) and 85.5mM cells were similar. The lack of inhibition of biomass accumulation, of all tested saline conditions clearly showed that the level of NaCl concentration used was not toxic for the cell metabolism. Also its ability to resist to 85.5mM NaCl can be on evidence that this suspension cel culture might have salt tolerance characteristics

Physical–chemical parameters and validation of a colorimetric method for deoxycholic and ursodeoxycholic acids: kit reagent and optical sensor

AbstractThe simple and low cost β-cyclodextrin (β-CD)–phenolphthalein (PHP) inclusion complex was used for both the study of physical–chemical parameters and validation of analytical procedures for deoxycholic acid (DCA) and ursodeoxycholic acid (UDCA) determinations in different formulations. The usefulness of this inclusion complex is proposed either in the form of kit reagent and as an original optical sensor for DCA and UDCA. The results showed that temperature had a negative effect on the equilibrium constant resulting in high negative values of enthalpy and positive values of entropy. The half-life values for DCA and UDCA measurements were 68.71 and 294.71 days, respectively. The method was validated showing limits of detection and quantification of 4.92×10−5molL−1 and 1.64×10−4molL−1 for DCA, 1.14×10−5molL−1 and 3.79×10−5molL−1 for UDCA, respectively. The developed optical sensor also showed response linearity, ease of implementation and potential application in fast screening tasks even out of the laboratory

Automatic miniaturized flow methodology with in-line solid-phase extraction for quinine determination in biological samples

The present work describes an analytical platform based on a multipumping flow injection analysis (MPFS) technique combined with in-line solid-phase extraction (SPE). The flow network has been tested with the determination of quinine in biological samples using fluorometry as the detection technique. Amberlite XAD-4 resin has been used as a solid phase and the implementation of a pH control strategy resulted in a simple and environmental approach for the preconcentration of quinine. Two solenoid valves allowed the deviation of the flow towards the resin column to carry out SPE procedures. The influence of parameters such as concentration, flow rate and volume of the different solutions on the sensitivity and performance of the MPFS was studied. Dynamic calibration ranges (0.78–150 ng mL 1) for quinine determination were applied by using a variable sample volume (120– 1000 mL). The developed methodology provided high relative extraction recoveries from human urine samples (85–115%). The proposed automatic methodology turns out to be very efficient and sustainable compared to the available procedures and it could prove to be an attractive alternative tool to perform in-line sample pre-treatment and subsequent direct determination of relevant organic compounds in pharmaceutical and clinical analyses

The microbial culture collections of the Federal University of Pernambuco (UFPE) and the new consortium towards the establishment of BRC-UFPE

The UFPE from Recife in Brazil hosts a bacterial (UFPEDA) and a fungal (URM) collections since 1951 and 1954, respectively. The UFPEDA was established by Prof. Oswaldo Gonçalves de Lima and is register in WDCM as 114. It is hosted at Antibiotic Department (DA) of UFPE and started out with 200 species mainly of the genus Streptomyces. Nowadays this collection holds 4000 strains of actinomycetes isolated from all the Brazilian places and from the International Streptomyces Project (ISP). The URM – University of Recife Mycology was established by Prof. Augusto Chaves Batista and is register in WDCM as 604. Actual it holds 9000 identified species including 1400 yeasts and 7600 filamentous fungi. All major fungal taxonomic groups are cover by this collection. The collections preserve each strain at least by two different techniques. Water and mineral oil storage were used for long operation time while freeze-drying and freezing at -80 ºC become the main techniques used at this stage. Special care is taken to test whether cultures recovered from preserved material conform to the original deposit. These collections have a range of services which are acceptance of free and confidential deposits, supply strains for academia, industry and services, support research and education (graduate and post-graduate students, as well as advanced training courses), identification services and confidential contracts (e.g. fungal medical diagnosis, starters for agro-industry companies, etc.). The OECD initiative related to guidance for the operation of Biological Resource Centres (BRC) is now a key reference for these collections. The right management of biological resources and their associate information including quality control are perused by these collections. The recent national projects, with reasonable budgets to support their activities, either on networking activities or requalification and management create a new breath and responsibilities to these collections. Taking advantage of good and well equipped premises of LIKA these collections are now open new avenues working in consortium to improve the quality control of their holdings using new tools from molecular biology and spectral analysis (MALDI-TOF) to achieve in the future a certified BRC for the UFPE microbial culture collections

Production and characterization of protease from Penicillium aurantiogriseum URM 4622

Proteases with new properties are required due to their increasing industrial importance. In this work, the optimal fermentation conditions for the production of a protease from Penicillium aurantiogriseum dierchx (URM-4622) are presented together with partial characterization of the protease catalytic properties. The batch fermentation conditions that allow for the highest specific proteolytic activity are 26 ºC, pH 7.0, and 25 % saturation dissolved O2 concentration. The obtained protease is stable over a wide range of pH (5.8 to 9.5) and temperature (25 to 40 ºC) values. In the presence of Zn2+ a 26 % reduction in the enzyme proteolytic activity occurs and, in contrast, Mn2+ enhances its activity by 28.9 %. 96.2 % and 70.8 % of the protease activity are maintained after 90 min incubation in 5 and 10 % (v/v) H2O2 aqueous solutions, respectively. PMSF inhibition reveals that this enzyme is a serine protease. Protease is able to hydrolyze different proteins

Multicommuted flow system for the determination of glucose in animal blood serum exploiting enzymatic reaction and chemiluminescence detection

An automatic flow procedure based on multicommutation dedicated for the determination of glucose in animal blood serum using glucose oxidase with chemiluminescence detection is described. The flow manifold consisted of a set of three-way solenoid valves assembled to implement multicommutation. A microcomputer furnished with an electronic interface and software written in Quick BASIC 4.5 controlled the manifold and performed data acquisition. Glucose oxidase was immobilized on porous silica beads (glass aminopropyl) and packed in a minicolumn (15 × 5 mm). The procedure was based on the enzymatic degradation of glucose, producing hydrogen peroxide, which oxidized luminol in the presence of hexacyanoferrate(III), causing the chemiluminescence. The system was tested by analysing a set of serum animal samples without previous treatment. Results were in agreement with those obtained with the conventional method (LABTEST Kit) at the 95% confidence level. The detection limit and variation coefficient were estimated as 12.0 mg l(−1) (99.7% confidence level) and 3.5% (n = 20), respectively. The sampling rate was about 60 determinations h(−1) with sample concentrations ranging from 50 to 600 mg l(−1) glucose. The consumptions of serum sample, hexacyanoferrate(III) and luminol were 46 μl, 10.0 mg and 0.2 mg/determination, respectively

Endoscopic closure of transmural bladder wall perforations

Background: Traditionally, intraperitoneal bladder perforations caused by trauma or iatrogenic interventions have been treated by open or laparoscopic surgery. Additionally, transvesical access to the peritoneal cavity has been reported to be feasible and useful for natural orifice translumenal endoscopic surgery (NOTES) but would be enhanced by a reliable method of closing the vesicotomy. Objective: To assess the feasibility and safety of an endoscopic closure method for vesical perforations using a flexible, small-diameter endoscopic suturing kit in a survival porcine model. Design, setting, and participants: This pilot study was performed at the University of Minho, Braga, Portugal, using six anesthetized female pigs. Interventions: Closure of a full-thickness longitudinal incision in the bladder dome (up to 10 mm in four animals and up to 20 mm in two animals) with the endoscopic suturing kit using one to three absorbable stitches. Measurements: The acute quality of sealing was immediately tested by distending the bladder with methylene-blue dye under laparoscopic control (in two animals). Without a bladder catheter, the animals were monitored daily for 2 wk, and a necropsy examination was performed to check for the signs of peritonitis, wound dehiscence, and quality of healing. Results and limitations: Endoscopic closure of bladder perforation was carried out easily and quickly in all animals. The laparoscopic view revealed no acute leak of methylene-blue dye after distension of the bladder. After recovery from anaesthesia, the pigs began to void normally, and no adverse event occurred. Postmortem examination revealed complete healing of vesical incision with no signs of infection or adhesions in the peritoneal cavity. No limitations have yet been studied clinically. Conclusions: This study demonstrates the feasibility and the safety of endoscopic closure of vesical perforations with an endoscopic suturing kit in a survival porcine model. This study provides support for further studies using endoscopic closure of the bladder which may lead to a new era in management of bladder rupture and adoption of the transvesical port in NOTES procedures

Antioxidant, phase II and III responses induced by lipoic acid in the fish Jenynsia multidentata (Anablapidae) and its influence on endolsulfan accumulation and toxicity

Antioxidants like lipoic acid (LA) are known to trigger augmented antioxidant and phase II and III responses. This study aimed to evaluate the effect of LA in P-glycoprotein (Pgp) expression, glutathione-S-transferase (GST) activity, total antioxidant competence, levels of lipid peroxides (TBARS) and accumulation of the organochlorine insecticide endosulfan (Endo: α-, β-isomers and sulfate metabolite) in different organs of the fish Jenynsia multidentata. One hundred and twenty females (1.55 ± 0.07 g) were fed during 8 days with (n = 60) or without (n = 60) a LA enriched ration (6000 mg/kg). Four experimental groups were defined: −LA/−Endo; +LA/−Endo; −LA/+Endo; and +LA/+Endo. Endo groups were exposed during 24 h to 1.4 μg of insecticide/L. Results showed that only LA induced a significant increment in liver Pgp expression. GST activity was augmented in liver after exposure to LA or Endo. TBARS levels were lowered in liver and gills after LA pre-treatment. Total antioxidant capacity was lowered in liver of Endo exposed fish, a result that was reversed by LA pre-treatment. It is concluded that LA induced the expected effects in terms of Pgp expression, GST activity and reduced TBARS levels although favored α-Endo accumulation in brain. However, the Endo metabolism to the more persistent endosulfan sulfate was not facilitated by LA pre-treatment.Fil: Monserrat, José María. Universidade Federal do Rio Grande do Norte; Brasil. Instituto de Ciências Biológicas; BrasilFil: Garcia, M. L.. Universidade Federal do Rio Grande do Norte; Brasil. Instituto de Ciências Biológicas; BrasilFil: Ventura Lima, J.. Universidade Federal do Rio Grande do Norte; Brasil. Instituto de Ciências Biológicas; BrasilFil: Gonzalez, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Ballesteros, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Miglioranza, Karina Silvia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Amé, María Valeria. Universidad Nacional de Córdoba; ArgentinaFil: Wunderlin, Daniel Alberto. Universidad Nacional de Córdoba; Argentin