Adenosine Triphosphate Measurements Research Articles

Reduction of Warm Ischemia Using a Thermal Barrier Bag in Kidney Transplantation: Study in a Pig Model

BackgroundWith recent advances in surgical technologies, minimally invasive endoscopic and robot-assisted surgical procedures have been introduced. However, prolonged warm ischemic time of the kidneys remains a concern after the organ is removed from a donor and during transplantation into a recipient. We developed a Thermal Barrier Bag (TBB) to prevent warm ischemia during transplantation. To confirm the effectiveness of the TBB, adenosine triphosphate (ATP) activity in the kidney was measured during an ex vivo warming test. An ischemia model porcine kidney was also used as the donor kidney and placed into the TBB; thereafter, the change in temperature at the time of transplantation was examined. Main FindingsThe purse-like design of the TBB efficiently suppressed heat conduction. A simulation was conducted that allowed the calculation of organ heat transfer condition. In the ex vivo experiment, temperature increases were suppressed in the group whose kidneys were placed in the TBB (30 minutes after transplantation: with TBB = 30°C, without TBB = 35°C). ATP measurements showed that the residual rate was substantially higher in the TBB group (P = .056). Moreover, a temperature suppression effect was demonstrated during the renal transplantation experiment (30 minutes after transplantation: with TBB = 27°C, without TBB = 31°C). ConclusionThe ex vivo warming experiment demonstrated that use of TBB slows down the rate of ATP decay in fresh kidneys. In addition, when an ischemic model porcine kidney was placed into the TBB and the temperature change at the time of transplantation was measured, an in vivo temperature-suppressing effect was observed.

A novel method: using an adenosine triphosphate (ATP) luminescence-based assay to rapidly assess the biological stability of drinking water.

The rapid and credible evaluations of the microbial stability of a drinking water distribution system (DWDS) are of great significance for ensuring the safety of drinking water and predicting microbial pollution. Conventional biostability assessment methods mainly focus on bacterial regrowth or evaluation of the level of nutrients that support bacterial regrowth. However, such methods are time-consuming and have many limitations. An adenosine triphosphate (ATP) assay can rapidly measure all active microorganisms and is known to be a useful method to assess the microbial activity of drinking water. The measurement of ATP has been used for more than a decade in the field of drinking water research. This article reviews the application of an ATP luminescence-based method to assess the biostability of drinking water and discusses the feasibility of ATP measurement as a parameter for quickly evaluating this criterion. ATP measurement will help researchers and water managers better monitor the biological stability of drinking water from the source to the consumer's tap. This review covers the: (1) principle and application of the ATP measurement in drinking water quality assessment; (2) comparison of the merits and demerits of several methods for evaluating the biostability of drinking water; (3) discussions on using ATP measurement in evaluating biostability; and (4) improvements in the use of ATP measurement in evaluating biostability. At the end of this review, recommendations were given for better application of the ATP measurement as a parameter for monitoring the microbial quality of drinking water.

Development of a point-of-contact technique to measure adenosine triphosphate: A quality improvement study.

PurposePatients with heart failure with preserved ejection fraction (HFpEF) experience fatigue due to impaired myocardial bioenergetics. Cardiomyocyte function depends on the delivery of adenosine triphosphate (ATP), yet there is no convenient bedside method to measure ATP. The purpose of this study was to develop a point-of-contact measurement of ATP that can be used in a clinical setting.MethodsIn a laboratory setting, digital finger punctures were conducted using 5 μl and 10 μl of capillary blood placed into various amounts of water (H2O). After mixing the solution for 10 s, a Hygiena AquaSnapTM Free ATP probe was placed into the solution for 10 s for the detection of ATP. The probe was then placed into the Hygiena luminometer for 15 s, and a value in relative light units (RLU) was obtained.ResultsTest samples using 10 μl of blood diluted from 50 to 500 mls of H2O produced ATP readings of 10,000-7569 RLUs. Using 5 μl of blood in 375–900 ml of H2O decreased the ATP values to 6459-4189 RLUs. Dilutional volume sparing experiments were conducted with ATP standards to determine the concentration of ATP per RLUs.ConclusionPatients with HFpEF have increased metabolic demand and impaired myocardial bioenergetics. Thus, identifying a method to measure ATP that is quick and accurate is imperative to accurately assess cellular energy production in this population. Point-of-contact measures, such as ATP, are needed for precision-guided treatment. Data from this study provides the first step toward developing evidence for health policies related to managing fatigue.

Building and indoor environmental quality assessment of Nigerian primary schools: A pilot study

A total of 15 classrooms went through on-site assessments/inspections, including measurements of temperature (T), and concentrations of carbon monoxide (CO) and carbon dioxide (CO2 ). In addition, the level of surface biocontamination/cleaning effectiveness was assessed by measuring adenosine triphosphate (ATP) levels on students' desks. Based on the data, the quality of facilities in the buildings was low. Classroom occupancy exceeded ASHRAE 50 person/100m2 standard in all cases indicating overcrowding. However, concentrations of CO2 remained below 1000ppm in most classrooms. On the other hand, indoor T was above the recommended levels for thermal comfort in all classrooms. Maximum indoor CO was 6ppm. Median ATP concentrations on the desk tops were moderately high in all schools. The use of open incinerators and power generator sets near classrooms, which was suspected to be the main source of CO, should be discouraged. Improved hygiene could be achieved by providing the students access to functioning bathroom facilities and cafeteria, and by effective cleaning of high contact surfaces such as desks. Although ventilation seems adequate based on CO2 concentrations, thermal comfort was not attained especially in the afternoon during extreme sunlight. Therefore, installing passive and/or mechanical cooling systems should be considered in this regard.

Impact of carbon-based nutrient enhancement on biofiltration performance for drinking water treatment

Incorporation of a carbon-based nutrient enhancement strategy for drinking water biofiltration is an attractive option, especially for source waters which contain recalcitrant organics. This study compared biofilters that were operated in parallel and individually enhanced with amino acids (including alanine, phenylalanine, and tryptophan), inulin, and sucrose to increase biomass concentration and promote biodegradation of dissolved organic carbon (DOC) in the source water, including disinfection by-product (DBP) precursors. Biomass activity was characterized by measuring adenosine tri-phosphate (ATP), dissolved oxygen (DO) consumption, and through the use of laccase and esterase enzyme assays. Performance was evaluated in terms of headloss, turbidity, pH, DOC, UV254, and DBP formation potential (DBP FP). The introduction of carbon-based nutrients significantly increased biomass activity, where ATP values peaked at 976 ng/g of filter media, 853 ng/g, and 513 ng/g for amino acids, inulin, and sucrose-spiked biofilters, respectively, while a non-spiked control only reached 104 ng/g. DO utilization by the enhanced biofilters was significantly higher than the control, with a strong correlation between ATP and DO uptake observed for all filters (R2 > 0.74). Laccase and esterase enzyme activities of enhanced biofilters were also elevated (p > 0.05), suggesting greater biodegradation potential. Operational parameters such as headloss development and turbidity control were not impaired by carbon supplementation strategies or an increase in biomass concentration and activity. However, the enhancement strategy did not provide improvement in terms of source water carbon removal (DOC and UV254) or DBP FP when treated filters were compared to a control.

The contribution of biofilm to nitrogenous disinfection by-product formation in full-scale cyclically-operated drinking water biofilters

Biofiltration has been shown to be effective for disinfection by-product (DBP) precursor control, however few studies have considered its role in the potential formation of DBPs. Biofilm is composed of heterogeneous bacteria as well as extracellular polymeric substances (EPS). The objective of this study was to determine the contribution of biofilm-related materials such as EPS to form nitrogen-containing DBPs upon chloramination, and to determine the influence of cyclical (scheduled on-off) biofilter operation on DBP precursor removal. Biologically active media was sampled from a full-scale biofilter operating under cold-water conditions (3.6 ± 0.5 °C) and extracted using a cation exchange resin into a phosphate buffer solution. Biomass concentrations, as determined using adenosine triphosphate (ATP) measurements, remained stable at 298 ± 55 ng ATP/g media over the trial period. N-nitrosodimethylamine (NDMA) and haloacetonitrile (HAN4) formation potential (FP) tests conducted under uniform formation conditions (UFC) using extracted biofilm yielded 0.80 ± 0.27 ng NDMA/g media and 18.7 ± 3.3 ng dichloroacetonitrile (DCAN)/g media. Further analyses of extracted biofilm using fluorescence spectroscopy and liquid chromatography-organic carbon detection indicated the presence of proteins above 20 kDa and humic-like substances. Extracted proteins (93.5 ± 8.1 μg/g media) correlated well (R = 0.90) with UV 280 measurements, indicating that spectrophotometry may serve as a valuable tool to quantify proteins in extracted biofilms. While substances in biofilms can serve as NDMA and DCAN precursors, the full-scale cyclically-operated biofilter that was examined did not show release of NDMA precursors during start-up following stagnation periods of 6 h or more. These biofilters consistently removed 6.9 ± 4.3 ng/L of NDMA precursors; typical NDMA UFC-FP of biofilter effluent was 8.5 ± 2.6 ng/L.

Using discrete and online ATP measurements to evaluate regrowth potential following ozonation and (non)biological drinking water treatment

Water utilities must control microbial regrowth in the distribution system to protect public health. In this study, an adenosine triphosphate (ATP)-based biomass production potential test using indigenous bacterial communities were used to evaluate regrowth potential following ozonation with either biofiltration (BF) or sustained chlorination (SCl2). Two full-scale water treatment plants with different upstream processes (i.e., WTP-BF: ozonation, coagulation/flocculation, biofiltration, UV irradiation, chlorination; and WTP-SCl2: ozonation, chlorination, coagulation/flocculation, filtration, chlorination) were compared. Characterization of indigenous bacteria using 16S rRNA gene sequencing, qPCR, and cellular ATP (cATP) showed microbial diversity changes across treatment, biomass sloughing from biofilters (effluent cATP = 30 ± 1 ng/L), and disinfection by chlorine (cATP < 1 ng/L). For both WTPs, 14-day cumulative biomass production (CBPt = ∑t=0tATP(t)×Δt) was highest for ozonated water samples (CBP14 = 1.2 × 103–3.0 × 103 d ngATP/L). CBP further increased with increasing ozone dose due to production of more biodegradable carbon. Growth promotion by carbon was confirmed from the consumption of ozonation byproducts (carboxylic acids, aldehydes) and the increase in CBP (9.5 × 102–2.9 × 103 d ngATP/L) after addition of 50–300 μgC/L acetate. Ozone followed by sustained chlorination (WTP-SCl2) effectively controlled biomass growth across the treatment process (CBP14 <10 d ngATP/L). In contrast, ozone followed by biofiltration (WTP-BF) reduced regrowth potential by 30% (biofilter influent CBP14 = 1.3 × 103 d ngATP/L; biofilter effluent CBP14 = 9.3 × 102 d ngATP/L). After adding chlorine to the biofilter effluent, CBP14 was reduced to <10 d ngATP/L. Lastly, online ATP measurements confirmed the discrete measurements and improved identification of the cATP peak and growth phases of indigenous bacteria.

Parkinsonian Neurotoxins Impair the Pro-inflammatory Response of Glial Cells.

In the case of Parkinson’s disease (PD), epidemiological studies have reported that pesticide exposure is a risk factor for its pathology. It has been suggested that some chemical agents, such as rotenone and paraquat, that inhibit the mitochondrial respiratory chain (in the same way as the PD mimetic toxin 1-methyl-4-phenylpyridinium, MPP+) are involved in the development of PD. However, although the neurotoxic effect of such compounds has been widely reported using in vivo and in vitro experimental approaches, their direct effect on the glial cells remains poorly characterized. In addition, the extent to which these toxins interfere with the immune response of the glial cells, is also underexplored. We used mouse primary mixed glial and microglial cultures to study the effect of MPP+ and rotenone on glial activation, in the absence and the presence of a pro-inflammatory stimulus (lipopolysaccharide plus interferon-γ, LPS+IFN-γ). We determined the mRNA expression of the effector molecules that participate in the inflammatory response (pro-inflammatory cytokines and enzymes), as well as the nitric oxide (NO) and cytokine production. We also studied the phagocytic activity of the microglial cells. In addition, we evaluated the metabolic changes associated with the observed effects, through the measurement of adenosine triphosphate (ATP) production and the expression of genes involved in the control of metabolic pathways. We observed that exposure of the glial cultures to the neurotoxins, especially rotenone, impaired the pro-inflammatory response induced by LPS/IFN-γ. MPP+ and rotenone also impaired the phagocytic activity of the microglial cells, and this effect was potentiated in the presence of LPS/IFN-γ. The deficit in ATP production that was detected, mainly in MPP+ and rotenone-treated mixed glial cultures, may be responsible for the effects observed. These results show that the response of glial cells to a pro-inflammatory challenge is altered in the presence of toxins inhibiting mitochondrial respiratory chain activity, suggesting that the glial immune response is impaired by such agents. This may have relevant consequences for brain function and the central nervous system’s (CNS’s) response to insults.

Neural-network classification of cardiac disease from 31P cardiovascular magnetic resonance spectroscopy measures of creatine kinase energy metabolism

BackgroundThe heart’s energy demand per gram of tissue is the body’s highest and creatine kinase (CK) metabolism, its primary energy reserve, is compromised in common heart diseases. Here, neural-network analysis is used to test whether noninvasive phosphorus (31P) cardiovascular magnetic resonance spectroscopy (CMRS) measurements of cardiac adenosine triphosphate (ATP) energy, phosphocreatine (PCr), the first-order CK reaction rate kf, and the rate of ATP synthesis through CK (CK flux), can predict specific human heart disease and clinical severity.MethodsThe data comprised the extant 178 complete sets of PCr and ATP concentrations, kf, and CK flux data from human CMRS studies performed on clinical 1.5 and 3 Tesla scanners. Healthy subjects and patients with nonischemic cardiomyopathy, dilated (DCM) or hypertrophic disease, New York Heart Association (NYHA) class I-IV heart failure (HF), or with anterior myocardial infarction are included. Three-layer neural-networks were created to classify disease and to differentiate DCM, hypertrophy and clinical NYHA class in HF patients using leave-one-out training. Network performance was assessed using ‘confusion matrices’ and ‘area-under-the-curve’ (AUC) analyses of ‘receiver operating curves’. Possible methodological bias and network imbalance were tested by segregating 1.5 and 3 Tesla data, and by data augmentation by random interpolation of nearest neighbors, respectively.ResultsThe network differentiated healthy, HF and non-HF cardiac disease with an overall accuracy of 84% and AUC > 90% for each category using the four CK metabolic parameters, alone. HF patients with DCM, hypertrophy, and different NYHA severity were differentiated with ~ 80% overall accuracy independent of CMRS methodology.ConclusionsWhile sample-size was limited in some sub-classes, a neural network classifier applied to noninvasive cardiac 31P CMRS data, could serve as a metabolic biomarker for common disease types and HF severity with clinically-relevant accuracy. Moreover, the network’s ability to individually classify disease and HF severity using CK metabolism alone, implies an intimate relationship between CK metabolism and disease, with subtle underlying phenotypic differences that enable their differentiation.Trial registrationClinicalTrials.gov Identifier: NCT00181259.

Comparing bacterial, fungal, and human cell concentrations with rapid adenosine triphosphate measurements for indicating microbial surface contamination

The goal of this study was to test for associations between adenosine triphosphate (ATP) content and microbial concentrations on desk surfaces in school classrooms. ATP bioluminescence and quantitative polymerase chain reaction (qPCR) techniques were employed to measure total bacterial, fungal, and human cell concentrations on 66 high-traffic desks spread across 9 schools: 3 in Connecticut (CT) and 6 in Oklahoma (OK). In CT, 6 samples were taken from each desk, 1 precleaning and 5 postcleaning (after 30 minutes, 1 day, 3 days, 7 days, and 21 days). In OK, samples were taken immediately before and after cleaning each desk. Based on simple linear regression analyses, ATP values were good predictors of microbial concentrations (r = 0.8, P = .003) in both CT school postcleaning samples and OK pre- and postcleaning samples (r = 0.7, P = .00002). However, biomass reductions measured after cleaning were 1.5-2 times greater when measured by ATP than by qPCR (P = .007). Overall, ATP bioluminescence measurements correlate with qPCR-based surface measurements on school desks but may overestimate the physical removal of bacteria and fungi due to cleaning.

ATP measurement in seawater reverse osmosis systems: Eliminating seawater matrix effects using a filtration-based method

A direct method for measuring adenosine-triphosphate (ATP) in seawater was developed recently, in which commercial reagents are added directly to seawater. However, calibration is required if seawater quality changes (such as changes in salinity, pH, Mg2+, Fe3+) as the seawater matrix interferes with ATP measurement. In this research, a 0.1 μm filtration process is introduced to eliminate such interferences. In addition, a filter rinsing step with sterilized artificial seawater is proposed to eliminate interference of free ATP.The ATP-filtration method is fast (<5 min), reproducible (VC = 7%), six times more sensitive than the direct ATP-method and correlates (R2 = 0.72, n = 100) with intact cell concentration. Microbial ATP concentration measured using the ATP-filtration method and the ATP-direct method were comparable. Microbial ATP measured along the treatment train of a full-scale seawater reverse osmosis (SWRO) plant decreased from 530 in the raw seawater to 10 ng-ATP/L after pre-treatment and to 0.5 ng-ATP/L in the SWRO permeate. The method was also applied to monitor bacterial growth potential (BGP) across the pre-treatment train of a (pilot) seawater desalination plant, where the removal of BGP through the media filtration and ultrafiltration was 44% and 7%, respectively.

Single and joint effects of Zn and Cu to ATP pool and microbial recovery in continuous growth systems

AbstractBACKGROUNDFour parallel continuous stirred tank reactors were used to investigate the single and joint exposure of Zn and Cu to Arthrobacter sp. JM018, in terms of the impact on the adenosine triphosphate (ATP) pool and microbial tolerance.RESULTSATP, optical density (OD), and substrate concentration measurements indicated that Cu was more toxic than Zn under all conditions studied. The results showed that although both OD and ATP measurements individually reflected a decrease in microbial growth rate after the addition of metals for the reactors exposed to Cu, specific ATP (i.e. ATP/OD) showed only a temporary reduction followed by a monotonic return to pre‐exposure levels within 100 h. The latter implied tolerance and recovery in the energy status of these cells, after the initial shock due to the exposure to Cu.CONCLUSIONSpecific ATP is an important quantity to be considered as a measure of the activity or energy status of microbial biomass surviving after exposure to toxic metals or other unfavorable conditions. The study provides insights for maintaining metal‐tolerant microbial communities and to explore both quantitatively and experimentally the dynamics of ATP pool and microbial tolerance in metal‐contaminated environments. © 2018 Society of Chemical Industry

1151. A Safer, More Effective Method for Cleaning and Disinfecting GI Endoscopic Procedure Rooms

BackgroundHealthcare acquired infections are increasing. Current cleaning and disinfecting (C&D) methods subject staff to toxic chemicals and can be damaging to the facility. Hypochlorous acid (HOCl) is a disinfecting solution that is 80–200 times more effective than bleach in surface disinfection of bacteria yet is nontoxic to humans. The aim of this study was to determine whether HOCl is as effective as standard cleaning methods for C&D GI ambulatory surgery center (ASC) rooms as determined by ATP (adenosine triphosphate) measurements over a 2-week period.MethodsTwo similar GI ASCs, each with two procedure rooms, were studied. One ASC received postprocedure STANDARD C&D with quaternary ammonium compounds in nonwoven fabric for surface wiping of high touch areas followed by terminal benzalkonium chloride foam/spray on these areas. The second ASC received HOCl C&D using on-site freshly prepared HOCl, 1,000 ppm. Microfiber cloths semi-wet with HOCl were used for wiping surfaces for both C&D. In the HOCl rooms, after terminal manual wiping, misting with HOCl of the entire room was performed. Selected high touch area ATP testing was performed in all rooms before procedures in the AM and 10 minutes after terminal manual cleaning. In the HOCL rooms, testing was also performed 10 minutes after misting. High touch areas tested in each room included: endoscopic cart (three locations/cart), both gurney bed rails, computer mouse (two), working counters (two areas), light switch, door knob. ATP scores were compared within each site using analysis of means (ANOM).ResultsAfter terminal cleaning, the average ATP score in the HOCl CLEANING and DISINFECTING study arm was significantly lower than that for the STANDARD CLEANING and DISINFECTING rooms (P < 0.0017) (Figure 1). In evaluating the effect of the HOCl misting, the ATP scores in the HOCl rooms had a post cleaning, pre-misting average score of 2.7. The post misting average score was 1.7, showing that misting produced a further significant reduction (improvement) in ATP scores (P = 0.01).ConclusionHOCl cleaning and disinfection in GI ASCs is more effective than standard procedures in lowering ATP scores following endoscopic procedures in procedure rooms. HOCl terminal misting of the rooms further improves the cleaning and disinfecting results.Figure 1.Disclosures B. Overholt, HOCl Solutions: Shareholder, none to date.

Rapid detection of microbial contamination in UHT milk: practical application in dairy industry

Microbial quality of ultra-high temperature (UHT) milk is usually ascertained by a total bacterial count (TBC) cultivation. But this is time consuming, so there is a tendency to search for faster and simpler methods. We compared three instruments, focusing on shortening the detection time and their suitability for practical use in dairy plants. Two of them, RapiScreen Dairy 1000 and Promilite III, detect microbial contamination by measuring adenosine triphosphate bioluminescence; the third, GreenLight, is based on oxygen consumption analysis. In the laboratory experiments, samples of UHT milk, were spiked with low concentration of microorganisms and then the level of microbial contamination was evaluated using the above-mentioned instruments together with cultivation method as a control. The instruments were also applied in a dairy plant to test 182 real samples. All investigated methods determined microbial quality faster than the TBC, but in some cases false positive and false negative results were obtained. Therefore, precise testing including optimizing pre-incubation time for bacteria enrichment is needed prior to industrial use.

Using upstream oxidants to minimize surface biofouling and improve hydraulic performance in GAC biofilters

The combination of biological growth and particle loading can adversely affect hydraulic performance in drinking water biofilters. In this study, upstream oxidant addition was used to distribute biologically-derived filter clogging in granular activated carbon (GAC) biofilters. Oxidant penetration was assessed during pilot-scale operation and backwashing of dual media (GAC/sand) and multimedia (GAC/anthracite/sand) biofilters. Influent chlorine (HOCl), monochloramine (NH2Cl), and hydrogen peroxide (H2O2) residuals were optimized to react with the GAC surface in the upper portion of the filter media bed (depth < 0.5 m) to attenuate biomass development. As the oxidant residual was quenched by surface-mediated reaction with the filter media, biomass growth was promoted deeper in the filter bed (depth > 0.5 m). The oxidant-induced effects on biomass and hydraulic performance were monitored through measurements of adenosine triphosphate (ATP) and head loss accumulation at different media depths. Addition of oxidants (e.g., 0.6 mg Cl2/L HOCl) could decrease terminal head loss by 20% in dual media filters and 40% in multimedia filters. These hydraulic benefits were achieved without significantly affecting removal of assimilable organic carbon (AOC), total organic carbon (TOC), turbidity, and particle counts. Oxidant type, residual concentration, media type, media age, and media depth influenced the passage of oxidant residuals and distribution of filter biomass. When oxidants were added during backwashing, oxidant residual was quenched through the bed depth from a combination of reactions with GAC media and biofilm degradation. This attenuation of residual oxidant may prevent the oxidant residual from penetrating the entire bed depth, potentially compromising backwashing objectives.

Effects of Hepatic Ischemia–Reperfusion Injuries and NRF2 on Transcriptional Activities of Bile Transporters in Rats

BackgroundThe effect of hepatic ischemia–reperfusion injury (IRI) on bile transporter (BT) gene expression is unknown. We hypothesized that abnormal expression of BTs during hepatic IRI is dependent on nuclear factor erythroid 2–related factor 2 (NRF2), which contributes to the cholestasis after reperfusion. MethodsSham surgery and short (60 min) or long (90 min) periods of warm ischemia time (WIT) with or without reperfusion for 24 h were applied to wild-type Sprague–Dawley rats and Nrf2 knockout rats (n = 5 per group). At each stage of IRI, the serum levels of aminotransferase, total bilirubin, and bile acids were measured. In addition, hepatic tissue was sampled to determine the histologic score of IRI (Suzuki score), measure adenosine triphosphate (ATP), and identify the quantitative real-time polymerase chain reactions of BTs (Oatp1, Ntcp, Mrp2, Bsep, and Mrp3). ResultsIn short periods of WIT, BT expression increased during the ischemia stage and returned to the baseline after reperfusion. However, in long periods of WIT, BT expression did not increase after ischemia and decreased further after reperfusion. Short WIT did not increase BT expression in Nrf2 knockout animals. The level of BT expression was correlated with the Suzuki score, the serum levels of aminotransferase, bilirubin, and bile acids, and tissue ATP level. Stepwise multiple regression analysis derived equations to predict the Suzuki score (R2 = 76.8, P < 0.001), serum total bilirubin (R2 = 61.2, P < 0.001), and tissue ATP (R2 = 61.1, P < 0.001). ConclusionsShort WIT induces the transcriptional activities of BT, whereas long WIT depresses them, and the effect was blunted by Nrf2 knockout status. BT expression can be considered a surrogate marker for hepatic IRI.

The potent and selective cyclin-dependent kinases 4 and 6 inhibitor ribociclib (LEE011) is a versatile combination partner in preclinical cancer models.

Inhibition of cyclin-dependent kinases 4 and 6 (CDK4/6) is associated with robust antitumor activity. Ribociclib (LEE011) is an orally bioavailable CDK4/6 inhibitor that is approved for the treatment of hormone receptor–positive, human epidermal growth factor receptor 2–negative advanced breast cancer, in combination with an aromatase inhibitor, and is currently being evaluated in several additional trials. Here, we report the preclinical profile of ribociclib.When tested across a large panel of kinase active site binding assays, ribociclib and palbociclib were highly selective for CDK4, while abemaciclib showed affinity to several other kinases. Both ribociclib and abemaciclib showed slightly higher potency in CDK4-dependent cells than in CDK6-dependent cells, while palbociclib did not show such a difference. Profiling CDK4/6 inhibitors in large-scale cancer cell line screens in vitro confirmed that RB1 loss of function is a negative predictor of sensitivity. We also found that routinely used cellular viability assays measuring adenosine triphosphate levels as a proxy for cell numbers underestimated the effects of CDK4/6 inhibition, which contrasts with assays that assess cell number more directly. Robust antitumor efficacy and combination benefit was detected when ribociclib was added to encorafenib, nazartinib, or endocrine therapies in patient-derived xenografts.

Enhanced biofilm solubilization by urea in reverse osmosis membrane systems

Chemical cleaning is routinely performed in reverse osmosis (RO) plants for the regeneration of RO membranes that suffer from biofouling problems. The potential of urea as a chaotropic agent to enhance the solubilization of biofilm proteins has been reported briefly in the literature. In this paper the efficiency of urea cleaning for RO membrane systems has been compared to conventionally applied acid/alkali treatment. Preliminary assessment confirmed that urea did not damage the RO polyamide membranes and that the membrane cleaning efficiency increased with increasing concentrations of urea and temperature. Accelerated biofilm formation was carried out in membrane fouling simulators which were subsequently cleaned with (i) 0.01M sodium hydroxide (NaOH) and 0.1M hydrochloric acid (HCl) (typically applied in industry), (ii) urea (CO(NH2)2) and hydrochloric acid, or (iii) urea only (1340 g/Lwater). The pressure drop over the flow channel was used to evaluate the efficiency of the applied chemical cleanings. Biomass removal was evaluated by measuring chemical oxygen demand (COD), adenosine triphosphate (ATP), protein, and carbohydrate content from the membrane and spacer surfaces after cleaning. In addition to protein and carbohydrate quantification of the extracellular polymeric substances (EPS), fluorescence excitation−emission matrix (FEEM) spectroscopy was used to distinguish the difference in organic matter of the remaining biomass to assess biofilm solubilization efficacy of the different cleaning agents. Results indicated that two-stage CO(NH2)2/HCl cleaning was as effective as cleaning with NaOH/HCl in terms of restoring the feed channel pressure drop (>70% pressure drop decrease). One-stage cleaning with urea only was not as effective indicating the importance of the second-stage low pH acid cleaning in weakening the biofilm matrix. All three chemical cleaning protocols were equally effective in reducing the concentration of predominant EPS components protein and carbohydrate (>50% reduction in concentrations). However, urea-based cleaning strategies were more effective in solubilizing protein-like matter and tyrosine-containing proteins. Furthermore, ATP measurements showed that biomass inactivation was up to two-fold greater after treatment with urea-based chemical cleanings compared to the conventional acid/alkali treatment. The applicability of urea as an alternative, economical, eco-friendly and effective chemical cleaning agent for the control of biological fouling was successfully demonstrated.

Biomass Recovery Method for Adenosine Triphosphate (ATP) Quantification Following UV Disinfection

ABSTRACTA biomass recovery method was developed to monitor UV disinfection efficacy using adenosine triphosphate (ATP). Typically, disinfection monitoring at wastewater treatment facilities (WWTFs) involves quantifying fecal and total coliforms or colony forming units, the results of which take a minimum of 24 h to produce. ATP quantification immediately before and after UV treatment, which takes only minutes, shows little reduction and often an increase in the microbial population since UV irradiation results in cells that are viable (i.e., still producing ATP) but not culturable. To overcome this, our biomass recovery method incorporates an incubation step to encourage life cycling of microbes. Average log reductions in cellular ATP (cATP) were found to be −0.28 ± 0.19, −0.011 ± 0.153, −0.17 ± 0.32, and 0.065 ± 0.074 using direct ATP measurements on UV-treated samples from WWTFs A, B, C, and D, respectively, while those using the recovery method were correspondingly 0.17 ± 0.34, 1.8 ± 0.8, 0.20 ± 0.35, and 0.72 ± 0.26. The response of the biomass recovery-ATP method indicated a significant direct correlation to the microbial population reduction observed in heterotrophic plate count (HPC) and Colilert® methods using both pure Escherichia coli culture and secondary municipal wastewater effluent.

Centrifugation-induced release of ATP from red blood cells.

Centrifugation is the primary preparation step for isolating red blood cells (RBCs) from whole blood, including for use in studies focused on transduction of adenosine triphosphate (ATP), an important vasodilatory signaling molecule. Despite the wide use of centrifugation, little work has focused on how the centrifugation itself affects release of ATP from RBCs prior to subsequent experimentation. Here we report that both the centrifugation force and duration have a pronounced impact on the concentration of ATP present in the packed RBCs following centrifugation. Multiple subsequent centrifugations yield extracellular ATP concentrations comparable to the amount released during the initial centrifugation, suggesting this effect is cumulative. Pairwise measurements of hemoglobin and ATP suggest the presence of ATP is primarily due to an increase in centrifugation-induced hemolysis. These results indicate that common centrifugation parameters, within the ranges explored here, can release ATP in quantities comparable to the low end of the range of values measured in typical ATP transduction experiments, potentially complicating experimental interpretation of those results.