A Laboratory Method for Determining Bacterially Formed Odorants and Reducing Odor in Absorbent Incontinence Products.

PURPOSE: The purpose of this study was to design a laboratory test method to mimic the formation of bacterially formed odorants during the use of absorbent urinary incontinence products. Three odor inhibitors with different modes of action were tested and evaluated. METHODS: Bacterially formed odorants in incontinence products were evaluated by adding a synthetic urine inoculated with a mixture of 4 bacterial strains to product samples cut from the incontinence products. The product samples were incubated in sealed flasks. The odorants that formed in the head space were sampled onto adsorbent tubes and analyzed by gas chromatography. The inhibitory effects of low pH, ethylenediaminetetraacetic acid (EDTA), and activated carbon were then measured. RESULTS: This technique enabled production of known odorants 3-methylbutanal, guaiacol, diacetyl, and dimethyl disulfide (DMDS) in concentrations of 50 to 600 ng/L in incontinence products. The method was further evaluated by testing 3 types of odor inhibitors; EDTA significantly reduced formation of all 4 odorants (P < .001). Lowering the pH from 6.0 to 4.9 decreased levels of 3-methylbutanal, DMDS, and guaiacol (P < .001); however, diacetyl levels increased (P < .001). Activated carbon significantly reduced the formation of diacetyl, DMDS, guaiacol, and 3-methylbutanal (P < .001). CONCLUSIONS: The technique we developed can be used to evaluate inhibitors with different modes of action to determine odor control in incontinence products. The odorants formed are produced by bacteria and have been identified as key contributors to the odor of used incontinence products. This work can be a step toward establishing a standard in the field of incontinence and odor control; creation of a standard will help the health care sector compare products to be purchased and benefit patients through the development of better products.

Scanning electroporation of selected areas of adherent cell cultures.

We present a computer-controlled scanning electroporation method. Adherent cells are electroporated using an electrolyte-filled capillary in contact with an electrode. The capillary can be scanned over a cell culture and locally deliver both an electric field and an electroporation agent to the target area without affecting surrounding cells. The instantaneous size of the targeted area is determined by the dimensions of the capillary. The size and shape of the total electroporated area are defined by these dimensions in combination with the scanning pattern. For example, striped and serpentine patterns of electroporated cells in confluent cultures can be formed. As it is easy to switch between different electroporation agents, the method is suitable for design of cell cultures with complex composition. Finite element method simulations were used to study the spatial distributions of the electric field and the concentration of an electroporation agent, as well as the fluid dynamics related to scanning and flow of electroporation agent from the capillary. The method was validated for transfection by introduction of a 9-base-pair-long randomized oligonucleotide into PC12 cells and a pmaxGFP plasmid coding for green fluorescent protein into CHO and WSS cells.

Generation of focused electric field patterns at dielectric surfaces.

We here report on a concept for creating well-defined electric field gradients between the boundaries of capillary electrode (a capillary of a nonconducting material equipped with an interior metal electrode) outlets, and dielectric surfaces. By keeping a capillary electrode opening close to a boundary between a conducting solution and a nonconducting medium, a high electric field can be created close to the interface by field focusing effects. By varying the inner and outer diameters of the capillary, the span of electric field strengths and the field gradient obtained can be controlled, and by varying the slit height between the capillary rim and the surface, or the applied current, the average field strength and gradient can be varied. Field focusing effects and generation of electric field patterns were analyzed using finite element method simulations. We experimentally verified the method by electroporation of a fluorescent dye (fluorescein diphosphate) into adherent, monolayered cells (PC-12 and WSS-1) and obtained a pattern of fluorescent cells corresponding to the focused electric field.

Single-cell electroporation.

Electroporation is a widely used method for the introduction of polar and charged agents such as dyes, drugs, DNA, RNA, proteins, peptides, and amino acids into cells. Traditionally, electroporation is performed with large electrodes in a batch mode for treatment of a large number of cells in suspension. Recently, microelectrodes that can produce extremely localized electric fields, such as solid carbon fiber microelectrodes, electrolyte-filled capillaries and micropipettes as well as chip-based microfabricated electrode arrays, have proven useful to electroporate single cells and subcellular structures. Single-cell electroporation opens up a new window of opportunities in manipulating the genetic, metabolic, and synthetic contents of single targeted cells in tissue slices, cell cultures, in microfluidic channels or at specific loci on a chip-based device.