Voiding camp: A successful and unique bladder rehabilitation program for children with urinary incontinence.

INTRODUCTION & BACKGROUND: Standard urotherapy is a well-established treatment for children with incontinence, although it is often challenging for both child and parents, and not always successful. As an alternative, several in- and outpatient bladder training programs have shown positive results on achieving continence. However, the disadvantage is the hospital environment, which can be more stressful for the child, and also quite expensive for society. OBJECTIVE: The aim was to evaluate the outcome on achieving continence following a voiding camp, where standard urotherapy was applied during a one-week stay at a regular summer youth camp, outside the hospital. STUDY DESIGN: Retrospective analysis of 105 children with urinary incontinence, followed in an expert centre for urinary incontinence for at least one year. Data at 7 different time points, before, during and until 6 months after voiding camp were collected. RESULTS: Even though all children had regular follow-up in an expert centre for urinary incontinence for at least one year before participating voiding camp, only 15% of the children reached the recommended amount of daily fluid intake (1.5 L/day). Once minimal daily fluid intake was re-established during the voiding camp, an immediate increase in the maximum voided volume (MVV), and a decrease in the number of wet days and wet nights per week was noted. This effect on a higher MVV remained even 3 months after voiding camp. DISCUSSION: Although sufficient daily fluid intake is a well-established part of standard urotherapy, up until now there was no data that proved the positive impact of sufficient daily fluid intake on bladder volume training and achieving continence in children. CONCLUSION: Voiding camp, as an unique bladder rehabilitation program for children with incontinence, is a successful alternative treatment option. Optimizing the daily fluid intake during voiding camp had a major positive impact on bladder volume training and achieving continence in children.

Patient information video: How to take care of your child after hypospadias surgery?

INTRODUCTION: Patient/parent education and participation helps improve post-operative care. Dressing and catheter care after hypospadias surgery varies widely and young parents are keen to use available media when seeking for help, especially if surgery is done in an outpatient setting. An information video about post-operative care after hypospadias is made available through a tertiary referral hospital's website. MATERIALS AND METHODS: Hypospadias surgery is an outpatient surgery in our setting. A double diaper system is used to prevent contamination of the urinary catheter and penile bandage with stools. A video explaining how the dressing works was made, helping parents maintaining the dressing and thereby solving possible questions/problems about dressing or medications while at home. Warning signs and symptoms are mentioned in which case parents should contact the urologist: fever, continuous blood loss and lack of urinary output. RESULTS: An information folder summarizing the latter, including a link to the video is given to all parents pre-operatively, providing reassurance according to parents' feedback. CONCLUSION: Parent participation can help improve post-operative care, especially in outpatient clinic setting. This video helps parents through post-operative care after hypospadias surgery. Its availability through informative folders and hospital's website is destined to educate and reassure parents.

e-Surveillance in animal health: use and evaluation of mobile tools.

In the last decade, mobile technology offered new opportunities and challenges in animal health surveillance. It began with the use of basic mobile phones and short message service (SMS) for disease reporting, and the development of smartphones and other mobile tools has expanded the possibilities for data collection. These tools assist in the collection of data as well as geo-referenced mapping of diseases, and mapping, visualization and identification of vectors such as ticks. In this article we share our findings about new technologies in the domain of animal health surveillance, based on several projects using a wide range of mobile tools, each with their specific applicability and limitations. For each of the tools used, a comprehensive overview is given about its applicability, limitations, technical requirements, cost and also the perception of the users.The evaluation of the tools clearly shows the importance of selecting the appropriate tool depending on the envisaged data to be collected. Accessibility, visualization and cost related to data collection differ significantly among the tools tested. This paper can thus be seen as a practical guide to the currently available tools.

Cerulenin inhibits de novo sophorolipid synthesis of Candida bombicola.

Synthesis of medium-chain sophorolipids by Candida bombicola is a challenging objective. One of the difficulties is that the obtained sophorolipids always represent a mixture of medium-chain and native de novo formed or long-chain sophorolipids. The fatty acid moiety of de novo sophorolipids is derived from the de novo synthesis of fatty acids. Fatty acid synthesis can be blocked by the antifungal agent cerulenin, an inhibitor if the fatty acid synthase (FAS) complex acting on the beta-ketoacyl thioester synthetase reaction. The toxic effect of cerulenin on C. bombicola was evaluated and 20 mg/ml was added in the stationary growth phase. No de novo formed sophorolipids were observed when the cells were cultured on merely glucose. Also when the hydrophilic substrate, 1,12-dodedanediol, was added, no de novo formed sophorolipids were detected, leading to a reduced complexity of the sophorolipid mixture.

Rapid isolation of fungal genomic DNA suitable for long distance PCR.

A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability for PCR. For Myrothecium gramineum, the highest DNA concentration was obtained with the procedure described by N. Vanittanakom et al. (J Clin Microbiol 2002, 40: 1739-1742). For A. nidulans, concentrations higher than 100 ng/mul were reached with the glass bead, the LiCl, the boiling, the liquid N(2) and the protoplast-based method. Samples of M. gramineum resulting from the boiling and the liquid N(2) procedure were suitable for the amplification of fragments up to 2.3 kb. The direct use of mycelium from M. gramineum in the PCR tube can be employed for the reproducible amplification of fragments up to 1 kb. Amplification of fragments up to 4.3 kb requires the use of the Elongase Mix on samples extracted with the liquid N(2) procedure.

Metabolic characterisation of E. coli citrate synthase and phosphoenolpyruvate carboxylase mutants in aerobic cultures.

E. coli is still one of the most commonly used hosts for protein production. However, when it is grown with excess glucose, acetate accumulation occurs. Elevated acetate concentrations have an inhibitory effect on growth rate and recombinant protein yield, and thus elimination of acetate formation is an important aim towards industrial production of recombinant proteins. Here we examine if over-expression of citrate synthase (gltA) or phosphoenolpyruvate carboxylase (ppc) can eliminate acetate production. Knock-out as well as over-expression mutants were constructed and characterized. Knocking out ppc or gltA decreased the maximum cell density by 14% and increased the acetate excretion by 7%, respectively decreased it by 10%. Over-expression of ppc or gltA increased the maximum cell dry weight by 91% and 23%, respectively. No acetate excretion was detected at these increased cell densities (35 and 23 g/l, respectively).

Transport kinetics of ectoine, an osmolyte produced by Brevibacterium epidermis.

Brevibacterium epidermis DSM 20659 is a halotolerant Gram-positive bacterium which can synthesize the osmolyte, ectoine, but prefers to take it up from its environment. The present study revealed that B. epidermis is equipped with at least one transport system for ectoine, with a maximal transport velocity of 15.7 +/- 4.3 nmol/g CDW.min. The transport requires energy (ATP) and is completely inhibited by the proton uncoupler, CCCP. The ectoine uptake system is constitutively expressed at a basal level of activity and its activity is immediately 10-fold increased by hyper-osmotic stress. Initial uptake rates are not influenced by the intensity of the hyper-osmotic shock but the duration of the increased activity of the uptake system could be directly related to the osmotic strength of the assay solution. Competition assays indicate that betaine, but not proline, is also transported by the ectoine uptake system.

Dynamics and optimal conditions of intracellular ectoine accumulation in Brevibacterium sp.

The optimal conditions for the intracellular synthesis of ectoine were determined in a halotolerant Brevibacterium sp. The size of the intracellular ectoine pool in the bacterial cells is shown to depend on the external salt concentrations, type of carbon source and aeration level. In erlenmeyer flasks a maximum concentration of intracellular ectoine of about 0.9 g/l was obtained. Under controlled aeration in a 1.5 l fermentor this level could be increased to 1.2 g/l. Consecutive cell transfers to media with increasingly higher salt concentrations enabled us to reach even higher levels, up to 1.6 g/l on erlenmeyer scale. The ectoine synthesis takes place immediately after the osmotic upshock. Within one generation time, the new corresponding specific intracellular ectoine concentration is reached.

Application of NAD-dependent polyol dehydrogenases for enzymatic mannitol/sorbitol production with coenzyme regeneration.

D-Mannitol and D-sorbitol were produced enzymatically from D-fructose using NAD-dependent polyol dehydrogenases. For the production of D-mannitol the Leuconostoc mesenteroides mannitol dehydrogenase could be used. Gluconobacter oxydans cell extract contained however both mannitol and sorbitol dehydrogenase. When this cell extract was used, the reduction of D-fructose resulted in a mixture of D-sorbitol and D-mannitol. To determine the optimal bioconversion conditions the polyol dehydrogenases were characterized towards pH- and temperature-optimum and -stability. As a compromise between enzyme activity and stability, the bioconversion reactions were performed at pH 6.5 and 25 degrees C. Since the polyol dehydrogenases are NADH-dependent, an efficient coenzyme regeneration was needed. Regeneration of NADH was accomplished by formate dehydrogenase-mediated oxidation of formate into CO2.

Enzymatic conversion of the clavan exopolysaccharide by Streptomyces sp. YSDL-20.

A screening programme was set up to isolate microorganisms able to hydrolyse the complex biopolymer clavan produced by Clavibacter michiganensis subsp. michiganensis LMG 5604. This valuable exopolysaccharide is very rich in L-fucose (37.5% w/w), a rare sugar, used in the medical field (Vanhooren, 1999). A microorganism capable of depolymerizing the polymer may decrease the high viscosity during clavan batch fermentations and remove the limitations of the oxygen transfer and consequently increase the clavan yield. It could also release free L-fucose or L-fucose rich oligosaccharides. An actinomycete, designated YSDL-20, isolated from a soil sample, was able to depolymerize this biopolymer. Based on its morphology and molecular characteristics, this strain could only be identified as Streptomyces sp.. On clavan, this strain displays good growth (17.5 g DCW/l after 96 h of cultivation) characterized by filamentous growth during the earlier days of cultivation followed by sporulation after 4 days. The flow behaviour of the Clavibacter broth was characterized, the fermentation culture broth behaves as a pseudoplastic fluid. The viscosity of the culture broth as well as of the purified clavan EPS, decreases when lyophilised supernatant of Streptomyces sp. YSDL-20 was added, indicating clavanase action. The viscosity decreases by 26% when the Clavibacter culture broth was incubated during 18 h with the crude Streptomyces enzyme source, whereas a 82% viscosity drop was observed, when the purified clavan EPS (10 g/l) was incubated with the lyophilised Streptomyces supernatant for 5 h.

Optimization of exopolysaccharide production by Tremella mesenterica NRRL Y-6158 through implementation of fed-batch fermentation.

In liquid culture conditions, the yeast-like fungus Tremella mesenterica occurs in the yeast state and synthesizes an exopolysaccharide (EPS) capsule, which is eventually released into the culture fluid. It is composed of an alpha-1,3-D-mannan backbone, to which beta-1,2 side chains are attached, consisting of D-xylose and D-glucuronic acid. Potato dextrose broth (PDB) seemed to be an excellent medium for both growth of the yeast cells and synthesis of the EPS. This medium is composed solely of an extract of potatoes to which glucose was added. Yet an important disadvantage of this production medium is the presence of starch in the potato extract, since Tremella cells are not capable of metabolizing this component; furthermore, it coprecipitates upon isolation of the polymer [3]. In this respect, it was essential to remove the starch in order to achieve high polysaccharide production and recovery. A good method was the removal of starch through ultrafiltration of the PDB medium before inoculation of the strain. This resulted in an excellent starch-free medium in which other components essential for polysaccharide production were still present [3]. Through implementation of single and cyclic fed-batch fermentations with glucose feed, 1.6- and 2.2-fold increases in EPS yield were obtained, respectively. Lowering the carbon source level by using a cyclic fed-batch technique might decrease the osmotic effect of glucose or any catabolite regulation possibly exerted by this sugar on enzymes involved in EPS synthesis.