Evaluation of a microscale quantitative suspension test to determine the bactericidal and yeasticidal activity of glutaral - one step to improve sustainability in disinfectant testing.

Aims: To evaluate a newly developed microscale quantitative suspension test compared to the existing standard suspension test using determination of the bactericidal and yeasticidal activity of glutaral as one step to improve the sustainability of disinfectant testing. Methods: The testing principles of the quantitative suspension test according to VAH method 9 (comparable to EN 13727) was used as a standard suspension test using 8.0 mL product test solution, 1.0 mL organic load and 1.0 mL test suspension. In addition, a micro-scale suspension test was performed in 96-well plates with 160 µL product test solution, 20 µL organic load and 20 µL test suspension. S. aureus ATCC 6538, P. aeruginosa ATCC 15442 and C. albicans ATCC 10231 were test organisms. Glutaral was tested at concentrations of 0.05%, 0.1%, 0.2% and 0.3% with exposure times of 1, 5 and 15 min. Polysorbate 80 (30 g/L), lecithin (9 g/L), L-histidine (1 g/L) and glycine (10 g/L) were used as validated neutralizers. After serial dilution of the disinfectant-neutralizer-mixture, plates were incubated for 48 h at 36°C (bacteria) or 72 hours at 30°C (C. albicans) and colony forming units (cfu) counted. The lg reduction was calculated as the difference between the results of the water control and the disinfectant at the end of the exposure time. All experiments were done in triplicate under clean conditions. Means of lg reduction were compared with the unpaired t-test, p<0.05 was considered to be significant. Results: Sufficient bactericidal activity according the VAH test requirements of at least 5 lg was found with both methods in 16 data sets of 24 data sets in total, and insufficient bactericidal activity of less than 5 lg was found with both methods in 7 data sets. In one data set, the mean lg reduction was above 5 lg with the microscale method and <5 lg with the VAH method, with no significant difference between the data sets (p=0.3096; 0.2% glutaral, 1 min, P. aeruginosa). A sufficient yeasticidal activity of at least 4 lg was found with both methods in one data set, an insufficient yeasticidal activity of less than 4 lg was found with both methods in 8 data sets. With one exception, no significant differences were detected between the two methods below the efficacy threshold. Conclusions: The microscale quantitative suspension test proved to provide results similar to those of VAH method 9 when the bactericidal and yeasticidal activity of glutaralwas evaluated, with 32 out of 33 evaluations yielding consistent results in terms of efficacy. Its suitability should be confirmed with additional bacterial species, additional biocidal active substances and in other laboratories.

Antimicrobial Efficiency and Surface Interactions of Quaternary Ammonium Compound Absorbed on Dielectric Barrier Discharge (DBD) Plasma Treated Fiber-Based Wiping Materials.

The physicochemical interactions between alkyldimethylbenzylammonium chloride (ADBAC) as disinfectant and three commercial wiping materials made from 100% polyester (PET), 55%cellulose/45%PET (blend), and 100% cellulose were investigated after treatment with dielectric barrier discharge (DBD) plasma at atmospheric pressure. Wipe material type in terms of cellulose content, liquor ratio, and immersion time demonstrated a significant influence on the adsorption of ADBAC. The higher the content of cellulose in the material, the higher is the adsorption of ADBAC active ingredient. The antimicrobial tests confirm that the ADBAC adsorbed on pure cellulosic material is inactivated losing its bactericidal activity, while 100% PET and blend wipes showed good antimicrobial efficacy. XPS analysis demonstrates the strong interactions of ADBAC with the plasma-generated oxygen species in the polyester-containing wipes surface. Unexpectedly, plasma-treated blend wipe displays a reverse antimicrobial effect compared to untreated samples, performing better in Gram-negative bacteria. The best result was obtained in the plasma treated 100% polyester wipe showing an improvement of about 20% in Gram-positive bacteria and an excellent performance in Gram-negative ones. This method allows the unprecedented use of pure polyester as effective wiping material for surface disinfection eliminating the major drawback of pure polyester, its high hydrophobicity.

Chemical, Thermo-Mechanical and Antimicrobial Properties of DBD Plasma Treated Disinfectant-Impregnated Wipes during Storage.

Disinfectant-impregnated wipes are broadly used in hospitals, as an important approach for infection prevention and control. But their ageing performance has rarely been studied. Untreated and Dielectric Barrier Discharge (DBD) plasma pre-treated wiping materials made of nonwoven 100% polyester (W1), nonwoven 55% cellulose/45% polyester (W2) and woven cotton (W3) were impregnated with a quaternary ammonium compound solution (ADBAC) for 30 min, 3, 7, 15, and 30 days of storage time and characterized in term of chemical, thermo-mechanical and antimicrobial efficacy. X-ray photoelectron spectroscopy analysis on the plasma-treated polyester wipes demonstrates the incorporation of reactive oxygen species on the fiber surface. Laser scanning microscopy demonstrates the plasma etching effect in smoothing the surface of the cotton wipe reducing the adsorption of ADBAC. The result showed no significant changes in breaking force and elongation during storage for W1 and W2. However, plasma treatment affects W3 in weft direction reducing the force at break in water and ADBAC treated wipes. Dynamic mechanical analysis results show that ADBAC and plasma treatment have a significant influence in W1 and W3 viscoelastic properties improving the elastic response limiting the polymeric chains mobility and the non-elastic response due to the etching effect, respectively. Overall, the plasma pre-treatment of ADBAC-impregnated wipes is able to enhance the antimicrobial performance and the storage time of polyester-containing wipes.

Efficacy of disinfectant-impregnated wipes used for surface disinfection in hospitals: a review.

Background: "Ready-to-use" disinfecting wipes (also known as pre-impregnated disinfecting wipe) are broadly used in food industry and domestic situations. Their application in hospitals and healthcare centres for decontamination of medical devices and surfaces is steadily increasing because of their convenient implementation in practice and reliable performance. Beside their acceptable compliance and easy application, literature reported the disinfection failure due to the interaction between textile substrate and active ingredients, which can highly increase the risk of an infection outbreak. This review aims to call attention to the wide range of variables affecting the disinfectant-impregnated wipes' (DIWs) disinfection performances in hospitals. Methods: A systematic literature search based on the five categories i. wipes, ii. disinfectants, iii. Application methods, iv. interaction between wipes and active ingredients and v. wiping strategy which can possibly influence the disinfection effectiveness of DIWs was conducted by Google scholar. Studies regarding the efficacy evaluation of DIWs in clinical applications were also reviewed from the National Centre for Biotechnology Information database. Results: Variables that impact on the disinfection performance of disinfectant-impregnated wipes in surface disinfection in hospitals were summarised and critically discussed. In addition to the information, current disinfectant-impregnated wipes' decontamination efficacy test standards were reviewed, and different standards exposed some disadvantage in their testing design. Conclusion: Various parameters contribute to the impact of DIWs disinfection performance in practice. The interaction between disinfectant active ingredients and the wiping materials barricades their broad application in hospitals. More studies of the DIWs' disinfection efficacy in clinical practice are in need. Current standards evaluating the DIWs' efficacy are required to improve for more realistic condition simulation and differentiating between mechanical removal of inoculum from a surface and chemical inactivation of the test microbe.

Modified silica sol coatings for surface enhancement of leather.

The presented study reports on differently modified silica sols for coating applications on leather. Silica sols are prepared by acidic hydrolysis of tetraethoxysilane and modified by silane compounds with fluorinated and non-fluorinated alkylgroups. In contrast to many earlier investigations regarding sol-gel applications on leather, no acrylic resin is used together with the silica sols when applying on leather. The modified silica particles are supposed to aggregate after application, forming thus a modified silica coating on the leather substrate. Scanning electron microscopy investigation shows that the applied silica coatings do not fill up or close the pores of the leather substrate. However, even if the pores of the leather are not sealed by this sol-gel coating, an improvement of the water repellent and oil repellent properties of the leather substrates are observed. These improved properties of leather by application of modified silica sols can provide the opportunity to develop sol-gel products for leather materials present in daily life.

Identification of a novel A-kinase anchoring protein 18 isoform and evidence for its role in the vasopressin-induced aquaporin-2 shuttle in renal principal cells.

Arginine vasopressin (AVP) increases the water permeability of renal collecting duct principal cells by inducing the fusion of vesicles containing the water channel aquaporin-2 (AQP2) with the plasma membrane (AQP2 shuttle). This event is initiated by activation of vasopressin V2 receptors, followed by an elevation of cAMP and the activation of protein kinase A (PKA). The tethering of PKA to subcellular compartments by protein kinase A anchoring proteins (AKAPs) is a prerequisite for the AQP2 shuttle. During the search for AKAP(s) involved in the shuttle, a new splice variant of AKAP18, AKAP18delta, was identified. AKAP18delta functions as an AKAP in vitro and in vivo. In the kidney, it is mainly expressed in principal cells of the inner medullary collecting duct, closely resembling the distribution of AQP2. It is present in both the soluble and particulate fractions derived from renal inner medullary tissue. Within the particulate fraction, AKAP18delta was identified on the same intracellular vesicles as AQP2 and PKA. AVP not only recruited AQP2, but also AKAP18delta to the plasma membrane. The elevation of cAMP caused the dissociation of AKAP18delta and PKA. The data suggest that AKAP18delta is involved in the AQP2 shuttle.

Merlin links to the cAMP neuronal signaling pathway by anchoring the RIbeta subunit of protein kinase A.

The cAMP-protein kinase A (PKA) pathway, important in neuronal signaling, is regulated by molecules that bind and target PKA regulatory subunits. Of four regulatory subunits, RIbeta is most abundantly expressed in brain. The RIbeta knockout mouse has defects in hippocampal synaptic plasticity, suggesting a role for RIbeta in learning and memory-related functions. Molecules that interact with or regulate RIbeta are still unknown. We identified the neurofibromatosis 2 tumor suppressor protein merlin (schwannomin), a molecule related to the ezrin-radixin-moesin family of membrane-cytoskeleton linker proteins, as a binding partner for RIbeta. Merlin and RIbeta demonstrated a similar expression pattern in central nervous system neurons and an overlapping subcellular localization in cultured hippocampal neurons and transfected cells. The proteins were coprecipitated from brain lysates by cAMP-agarose and coimmunoprecipited from cellular lysates with specific antibodies. In vitro binding studies verified that the interaction is direct. The interaction appeared to be under conformational regulation and was mediated via the alpha-helical region of merlin. Sequence comparison between merlin and known PKA anchoring proteins identified a conserved alpha-helical PKA anchoring protein motif in merlin. These results identify merlin as the first neuronal binding partner for PKA-RIbeta and suggest a novel function for merlin in connecting neuronal cytoskeleton to PKA signaling.

The neuA/flmD gene cluster of Helicobacter pylori is involved in flagellar biosynthesis and flagellin glycosylation.

Helicobacter pylori possesses a gene (HP0326/JHP309) homologous to neuA of other bacteria, encoding a cytidyl monophosphate-N-acetylneuraminic acid synthetase-homologous enzyme in its N-terminal portion. We analysed the function of this gene, which is controlled by a flagellar class 2 sigma(54) promoter, in flagellar biosynthesis. HP0326/JHP309 actually represents a bicistronic operon consisting of a neuA and a flmD-like putative glycosyl transferase gene. An isogenic flmD mutant synthesized basal bodies but no filaments, was non-motile, and expressed severely reduced amounts of a FlaA flagellin of reduced molecular mass. FlaA flagellin was found to be glycosylated in its exported form within the flagellar filament, but not inside the cytoplasm. Glycosylated FlaA was not detectable in the flmD mutant. Together with other genes in the H. pylori genome, a proposed function of the neuA/flmD gene products could be to provide a pathway for glycosylation of flagellin and other extracytoplasmic molecules during type III secretion.