Calcium hypochlorite on mouse embryonic fibroblast cells (NIH3T3) in vitro cytotoxicity and genotoxicity: MTT and comet assay

Molecular Biology Reports - Antimicrobial irrigation solutions are widely used under clinical settings. Their effect on dental tissue is a subject of recent research, which aims for a safer...     

DIFFERENT RECOVERY METHODS AND MUSCLE PERFORMANCE AFTER EXHAUSTING EXERCISE: COMPARISON OF THE EFFECTS OF ELECTRICAL MUSCLE STIMULATION AND MASSAGE

In this study we assessed the influence of the three different recovery interventions massage (MSG), electrical muscle stimulation (EMS), and passive rest (PR) on lactate disappearance and muscle recovery after exhausting exercise bouts. Twelve healthy male sport students participated in the study. They attended the laboratory on five test days. After measurement of V.O₂max and a baseline Wingate test (WG_b), the three recovery interventions were tested in random counterbalanced order. High intensity exercise, which consisted of six exhausting exercise bouts (interspersed with active recovery), was followed by MSG, EMS or PR application (24 minutes); then the final Wingate test (WG_f) was performed. Lactate, heart rate, peak and mean power, rating of perceived exertion (RPE), and total quality of recovery (TQR) were recorded. In WG_f mean power was significantly higher than in WG_b for all three recovery modalities (MSG 6.29%, EMS 5.33%, PR 4.84% increase, p < 0.05), but no significant differences in mean and peak power were observed between the three recovery modes (p > 0.05). The heart rate response and the changes in blood lactate concentration were identical in all three interventions during the entire protocol (p = 0.817, p = 0.493, respectively). RPE and TQR scores were also not different among the three interventions (p > 0.05). These results provide further evidence that MSG and EMS are not more effective than PR in the process of recovery from high intensity exercise.     

The detection of <Emphasis Type="Italic">hip</Emphasis>O gene by real-time PCR in thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> spp. with very weak and negative reaction of hippurate hydrolysis

A total of 190 Campylobacter spp. isolates, of which 34 gave the result of very weak activity, and 156 gave the negative activity in the test for hippurate hydrolysis were characterized. The genomic DNA was isolated from a fresh culture of each isolate and the real-time PCR, targeting the hipO gene, was used to confirm the species distribution of Campylobacter isolates. The hipO gene was detected in 17 isolates (11%) within the total of 156 negative isolates for hippurate hydrolysis. Out of 34 isolates with very weak activity, 19 isolates (56%) were also found to be positive for hipO gene and characterized as C. jejuni. The real-time PCR assay used in this study could be employed for more accurate diagnosis of Campylobacter infections at species level after the biochemical characterization based on hippuricase activity of the isolates. This could also provide important data for the epidemiology of infections associated with these zoonotic pathogens.     

Barley mildew and its elicitor chitosan promote closed stomata by stimulating guard-cell S-type anion channels

Stomatal closure is known to be associated with early defence responses of plant cells triggered by microbe-associated molecular patterns (MAMPs). However, the molecular mechanisms underlying these guard-cell responses have not yet been elucidated. We therefore studied pathogen-induced changes in ion channel activity in Hordeum vulgare guard cells. Barley mildew (Blumeria graminis) hyphae growing on leaves inhibited light-induced stomatal opening, starting at 9 h after inoculation, when appressoria had developed. Alternatively, stomatal closure was induced by nano-infusion of chitosan via open stomata into the sub-stomatal cavity. Experiments using intracellular double-barreled micro-electrodes revealed that mildew stimulated S-type (slow) anion channels in guard cells. These channels enable the efflux of anions from guard cells and also promote K(+) extrusion by altering the plasma membrane potential. Stimulation of S-type anion channels was also provoked by nano-infusion of chitosan. These data suggest that MAMPs of mildew hyphae penetrating the cuticle provoke activation of S-type anion channels in guard cells. In response, guard cells extrude K(+) salts, resulting in stomatal closure. Plasma membrane anion channels probably represent general targets of MAMP signaling in plants, as these elicitors depolarize the plasma membrane of various cell types.     

A quantitative colorimetric method of measuring alkaline phosphatase activity in eukaryotic cell membranes

Alkaline phosphatase (ALP) is glycoprotein structured metalophosphatase with several defined functions. It is present in many tissues of all living beings from bacteria to mammals. The enzyme may catalyse the hydrolysis of various monophosphate esters at alkaline pH. The objective of this study was to quantify ALP functioning particularly in the membranes of eukaryotic cells. The membranes of seven different cells (myeloma cells; hybrid cells; erythroleukaemia cells; lymphocytes and erythrocytes) were tested for ALP activity using a cellular enzyme assay, which is based on the conversion of para-nitrophenylphosphate (p-NPP) to para-nitrophenol and the colorimetric determination of the resulting coloured product. The test system was optimised with respect to substrate concentration, reaction time and the number of cells used as a source of enzyme. The obtained values were converted to quantitative results through a standard curve created using commercial ALP. In order to determine the effect of serum concentration on enzyme activity, 1G2 hybridoma, which is among the cells used in this study and which synthesizes monoclonal antibody against human serum albumin, was produced in different serum concentrations ranging from 0 to 15%.     

Extremely low‐frequency electromagnetic fields affect the immune response of monocyte‐derived macrophages to pathogens

This study aimed to determine the effect of extremely low‐frequency electromagnetic fields (ELF‐EMF) on the physiological response of phagocytes to an infectious agent. THP‐1 cells (human monocytic leukemia cell line) were cultured and 50 Hz, 1 mT EMF was applied for 4–6 h to cells induced with Staphylococcus aureus or interferon gamma/lipopolysaccharide (IFγ/LPS). Alterations in nitric oxide (NO), inducible nitric oxide synthase (iNOS) levels, heat shock protein 70 levels (hsp70), cGMP levels, caspase‐9 activation, and the growth rate of S. aureus were determined. The growth curve of exposed bacteria was lower than the control. Field application increased NO levels. The increase was more prominent for S. aureus‐induced cells and appeared earlier than the increase in cells without field application. However, a slight decrease was observed in iNOS levels. Increased cGMP levels in response to field application were closely correlated with increased NO levels. ELF‐EMF alone caused increased hsp70 levels in a time‐dependent manner. When cells were induced with S. aureus or IFγ/LPS, field application produced higher levels of hsp70. ELF‐EMF suppressed caspase‐9 activation by a small extent. These data confirm that ELF‐EMF affects bacterial growth and the response of the immune system to bacterial challenges, suggesting that ELF‐EMF could be exploited for beneficial uses. Bioelectromagnetics 31:603–612, 2010. © 2010 Wiley‐Liss, Inc.     

Effects of short-time drying on biofilm-associated bacteria

Microorganisms tend to form biofilms consisting of cells embedded in a highly hydrated extracellular polymeric matrix. The biofilm protects its inhabitants from antimicrobial agents, pH alterations, and confers protection against drying. It is known that biofilm-associated bacteria can survive for a while in the absence of water. When rehydrated, metabolic processes are quickly restored and microorganisms resume life. The aim of this study is to evaluate the survival of heterotrophic bacteria, sulphate-reducing bacteria and amoeba against short-time drying. Biofilms were allowed to grow for 30 and 60 days on stainless steel (316, 2B) coupons in annular biofilm reactor, which was fed with drinking water network under constant, non-turbulent shear stress and temperature. The results presented in this study indicate a role for biofilm layer in protecting biofilm-associated microorganisms from drying. The current study has provided that short-time (24 h) absence of water could not affect biofilm-associated heterotrophic microorganisms significantly, in terms of cell viability.     

Effect of oxidative stress on in vivo ADP-ribosylation of eukaryotic elongation factor 2

Different lines of evidence indicate that eukaryotic elongation factor 2 (eEF2) can be ADP-ribosylated endogenously. The physiological significance of this reaction has, however, remained unclarified. In order to address this issue we investigated the in vivo ADP-ribosylation of eEF2 and the effect of oxidative stress thereon. The investigation revealed that the endogenous ADP-ribosylation of eEF2 is complex and can take place in K562 cell lysates either under the action of endogenous transferase from [adenosine-14C]NAD or by direct binding of free [14C]ADP-ribose. These two types of ADP-ribosylation were distinguished by use of different treatments based on the chemical stability of the respective bonds formed. Under standard culture conditions, in vivo labeling of eEF2 in the presence of [14C]adenosine was reversed to about 65% in the presence of diphtheria toxin and nicotinamide. This finding implied that the modification that took place under physiological circumstances was, mainly, of an enzymic nature. On the other hand, H2O2-promoted oxidative stress gave rise to a nearly two-fold increase in the extent of in vivo labeling of eEF2. This was accompanied by a loss of eEF2 activity in polypeptide chain elongation. Oxidative stress specifically inhibited the subsequent binding of free ADP-ribose to eEF2. The results thus provide evidence that endogenous ADP-ribosylation of eEF2 can also take place by the binding of free ADP-ribose. This nonenzymic reaction appears to account primarily for in vivo ADP-ribosylation of eEF2 under oxidative stress.     

Efficacy of a quaternary ammonium compound against planktonic and sessile populations of differentLegionella pneumophila strains

Efficacy of Gemacide PN-50^TM (a quaternary ammonium compound) as a commercial formulation recommended for disinfecting heat exchangers was determined for both planktonic and sessile populations of variousLegionella pneumophila strains. The quaternary ammonium compound (QAC) was preferred as an alternative due to the emerging resistance of potentially pathogenic bacteria against different biocides. PlanktonicL. pneumophila strains were suspended in tap water while sessile ones were grown on stainless steel that is used in construction of the cooling towers, then both group of strains were exposed to the biocide. The sensitivity of both planktonic and sessile populations ofL. pneumophila strains to the biocide was different. The biocide was found effective below recommended dosages (1000–2000 mg/L) against planktonic populations ofL. pneumophila, whereas it was determined that higher than the recommended dosages were required for sessile populations. The environmental isolates were more resistant to the biocide than the ATCC isolate was. The results indicated that studying only the planktonic populations ofL. pneumophila for biocide tests might not be sufficient to provide the optimum dosage and contact time information for field trials. Therefore, biocidal activity of a water treatment chemical must be evaluated in terms of dosage and contact times on both planktonic and sessile bacteria.