ATP Bioluminescence Assay Research Articles

Assessing effectiveness of cleaning and disinfection of equipment and environmental surfaces in cystic fibrosis clinics using an ATP assay.

Infection control guidelines for cystic fibrosis (CF) stress cleaning of environmental surfaces and patientcare equipment in CF clinics. This multicenter study measured cleanliness of frequently touched surfaces in CF clinics using an ATP bioluminescence assay to assess the effectiveness of cleaning/disinfection and the impact of feedback. Eight surfaces were tested across 19 clinics (10 pediatric, 9 adult) over 5 rounds of testing. Rounds 1 and 2 served as uncleaned baseline, and Round 3 occurring after routine cleaning. Rounds 4 and 5 were performed after feedback provided to staff and measured after cleaning. Pass rates defined as <250 relative light units were the primary outcome. Of the 750 tests performed, 72% of surfaces passed at baseline, and 79%, 83%, and 85% of surfaces passed in Rounds 3, 4, and 5, respectively. The overall pass-rate was significantly higher in adult compared to pediatric clinics (86% vs 71%; P < 0.001). In pediatric clinics, blood pressure equipment and computer keyboards in the pulmonary function lab consistently passed, but the exam room patient/visitor chairs consistently failed in all rounds. In adult clinics blood pressure equipment, keyboards in exam rooms and exam tables passed in all rounds and no surface consistently failed. We demonstrate the feasibility of an ATP bioluminescence assay to measure cleanliness of patient care equipment and surfaces in CF clinics. Pass rates improved after cleaning and feedback for certain surfaces. We found that surfaces are more challenging to keep clean in clinics taking care of younger patients.

Simultaneous determination of somatic cell count and total plate count in raw milk based on ATP bioluminescence assay

The somatic cell count (SCC) and total plate count (TPC) are essential quality indicators for raw milk. Traditional detection methods require separate measurements and rely on complex, large-scale instruments or cultivation techniques, which are both time-consuming and laborious. To address these challenges, this study developed a novel method for the simultaneous detection of SCC and TPC in the same raw milk sample using the ATP bioluminescence assay. This method utilizes oxy-ethylated iso-nonyl phenol (Neonol-10) and cetyltrimethylammonium bromide (CTAB) to selectively lyse somatic cells and microorganisms, respectively. This technique is straightforward to operate and can be completed within 2.5 h, with detection ranges of 1 × 10⁴ to 3 × 10⁶ cells/mL for SCC and 1 × 10⁵ to 5 × 10⁷ CFU/mL for TPC. Importantly, this technique meets the requirements of detection standards in China, European Union, Canada, United States, etc. For SCC or TPC in raw milk. Overall, this innovative approach does not rely on expensive equipment or facilities and the stepwise reagent addition procedure can be easily developed into an automated high-throughput system for rapid on-site testing of SCC and TPC in raw milk.

Hygiene Assessment of Buffalo Milking Parlours in Campania Region, Italy: A Preliminary Study by Using ATP Luminometry and Bacteriological Investigation.

Careful cleaning of a milking parlour and its equipment is fundamental to guarantee good raw milk quality and prevent the dissemination of bacteria and improve animal welfare. This study aimed to investigate, using an ATP-bioluminescence assay and bacteriological analysis, the bacterial contamination of milking parlours on milking parlour surfaces of buffalo farms in the Campania Region, evaluating the seasonal dynamics during the year 2022. Eight farms were selected by the Italian ClassyFarm system, which assesses the level of animal welfare and biosecurity according to risk analysis. Before sampling, all dairy farm owners filled out a questionnaire on milking management, animal hygiene, and health. The questionnaires evidenced similar cleaning procedures but an absence of a standardised cleaning protocol among the different farms. ATP bioluminescence results evidenced similar levels of contamination in all the selected buffalo farms, and the season comparison showed no significant differences. A variation in the percentages of bacterial isolates during the different seasons was observed, with a higher prevalence of Enterobacteriaceae (38%) in summer. A small number of samples exhibited an absence of bacterial growth. Identifying bacteria is crucial for understanding the microorganisms present in the milking parlour, yet employing ATP luminometry could offer broad and accurate applications in buffalo milking parlours. In conclusion, the use of ATP bioluminescence for evaluating the hygiene of a buffalo milking parlour could represent a further important advancement in dairy farming technology.

Shenqi Fuzheng injection facilitates skeletal muscle mitophagy mediated by the ubiquitination of HIF-1α to ameliorate cancer-associated fatigue.

Cancer-related fatigue (CRF) significantly impacts the quality of life of cancer patients. This study investigates the therapeutic potential of Shenqi Fuzheng injection (SFI) in managing CRF, focusing on its mechanistic action in skeletal muscle. We utilized a CRF mouse model to examine the effects of SFI on physical endurance, monitoring activity levels, swimming times and rest periods. Proteomic analysis of the gastrocnemius muscle was performed using isobaric tags and liquid chromatography-tandem mass spectrometry to map the muscle proteome changes post-SFI treatment. Mitochondrial function in skeletal muscle was assessed via ATP bioluminescence assay. Furthermore, the regulatory role of the hypoxia inducible factor 1 subunit alpha (HIF-1α) signalling pathway in mediating SFI's effects was explored through western blotting. In CRF-induced C2C12 myoblasts, we evaluated cell viability (CCK-8 assay), apoptosis (flow cytometry) and mitophagy (electron microscopy). The study also employed pulldown, luciferase and chromatin immunoprecipitation assays to elucidate the molecular mechanisms underlying SFI's action, particularly focusing on the transcriptional regulation of PINK1 through HIF-1α binding at the PINK1 promoter region. Our findings reveal that SFI enhances physical mobility, reduces fatigue symptoms and exerts protective effects on skeletal muscles by mitigating mitochondrial damage and augmenting antioxidative responses. SFI promotes cell viability and induces mitophagy while decreasing apoptosis, primarily through the modulation of HIF-1α, PINK1 and p62 proteins. These results underscore SFI's efficacy in enhancing mitochondrial autophagy, thereby offering a promising approach for ameliorating CRF. The study not only provides insight into SFI's potential therapeutic mechanisms but also establishes a foundation for further exploration of SFI interventions in CRF management.

Evaluation of the viability of the Bulgarian BCG vaccine by the adenosine triphosphate assay

We applied a modified assay for the measurement of the intracellular ATP content as an alternative biochemical method for estimating the viability of the BCG vaccine instead of the standard cultural microbiological assay. Eight lyophilized BCG ampoule samples (Lot 287-2 and Lot 254-2, Bulgarian BCG strain SL222 Sofia) were tested on different days. Each calibration curve was measured for the following range of ATP concentrations: 100, 10, 5, 2.5, 1.25, 0.625, 0.3125, and 0.1562 pmol. The obtained results were compared to the traditional cultural colony counting method for evaluating the viability of the BCG vaccine. The NCP of Lot 287-2 (min 2.325–max 3.300) was higher in comparison to Lot 254-2 (min 1.438–max 3.330). That result corresponded to higher ATP-content registered for Lot 287-2 (min 0.409–max 1.338) than Lot 254-2 (min 0.18–max 1.104). Our results confirmed the observation made by other researchers that the modified ATP assay is a reliable, simple, and quick method for the evaluation of the viability of the BCG vaccine. Additional compiled data about all BCG strains are needed to perform studies on the correlation between the luciferin-luciferase bioluminescent ATP assay and the cultural method for viability and enumeration of the units in the lyophilized BCG vaccine.

Abstract 7300: Synergistic inhibition of prostate cancer growth by the combination of xanthohumol and ursolic acid

Abstract Within the United States, prostate cancer (PCa) is the most commonly diagnosed non-cutaneous cancer, and approximately 1 in 8 men will receive a diagnosis in their lifetime. As the second leading cause of cancer deaths in American men, about 1 in 41 will die because of prostate cancer. The World Health Organization has suggested that approximately one-third of all cancer deaths are preventable, and one of the suggested means of prevention is through the increased consumption of fruits and vegetables. Fruits and vegetables contain natural compounds known as phytochemicals which possess biological activity including some anti-cancer effects. By adapting the ATP-bioluminescence assay for cell viability, our laboratory screened a natural compound library to find combinations of these compounds that may synergistically inhibit PCa cell viability. One of the combinations to emerge from this screening process was xanthohumol (XAN) and ursolic acid (UA). We examined the ability of the combination to inhibit tumor growth in a mouse HMVP2 PCa allograft tumor model. Mice were fed diets supplemented with either XAN (0.2%) or UA (0.2%) alone or the combination. Mice on the combination diet showed a significant inhibition of HMVP2 PCa tumor growth compared to mice on the single-agent diets. In addition, in vitro studies were performed to support the allograft tumor study and to further examine the mechanism of action of the combination. A cell survival assay utilizing six different PCa cell lines indicated that the combination of XAN and UA synergistically inhibited the survival of all cell lines as calculated by the Bliss index. Mechanistic studies performed in PCa cells indicate that the combination of XAN and UA inhibits oncogenic signaling better than either agent alone. Additionally, unbiased metabolomics revealed that the tumors from mice fed the combination diet had significantly less S-adenosyl-L-methionine compared to all other groups. Overall, 13 metabolic pathways were significantly different in the tumors from mice fed the combination diet compared to all other groups. The current data suggest that using a combination of XAN and UA may be a valuable chemopreventive strategy for PCa in humans. Funding: This work was supported by NIH Grant CA228404. Citation Format: Rachel Clark, Achinto Saha, Chelsea A. Friedman, Ruggiero Gorgoglione, Stefano Tiziani, John DiGiovanni. Synergistic inhibition of prostate cancer growth by the combination of xanthohumol and ursolic acid [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7300.

Application of BactTiter-Glo ATP bioluminescence assay for Mycobacterium tuberculosis detection

BackgroundTuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), remains a global health threat, necessitating faster and more accessible diagnostic methods. This study investigates critical parameters in the application of a commercial ATP bioluminescence assay for the detection of MTB. MethodOur objective was to optimize the ATP bioluminescence protocol using BacTiter-Glo™ for MTB, investigating the impact of varying volumes of MTB suspension and reagent on assay sensitivity, evaluating ATP extraction methods, establishing calibration curves, and elucidating strain-specific responses to antimicrobial agents. ResultsATP extraction methods showed no significant improvement over controls. Calibration curves revealed a linear correlation between relative light units (RLU) and colony-forming units (CFU/mL), establishing low detection limits. Antimicrobial testing demonstrated strain-specific responses aligning with susceptibility and resistance patterns. ConclusionOur findings contribute to refining ATP bioluminescence protocols for enhanced MTB detection and susceptibility testing. Further refinements and validation efforts are warranted, holding promise for more efficient diagnostic platforms in the future.

Immunohistochemical localization of P2Y12 purinoceptors in the rat carotid body

The present study investigated the localization of the adenosine 5′-diphosphate (ADP)-selective P2Y12 purinoceptors in the rat carotid body using multilabeling immunofluorescence. Punctate immunoreactive products for P2Y12 were distributed in chemoreceptive type I cells immunoreactive to vesicular nucleotide transporter (VNUT) or dopamine beta-hydroxylase, but not in S100B-immunoreactive glial-like type II cells. P2Y12 immunoreactivity was localized in cell clusters containing VNUT-immunoreactive type I cells surrounded by the perinuclear cytoplasm and cytoplasmic processes of type II cells immunoreactive for ectonucleoside triphosphate diphosphohydrolase 2 (NTPDase2) and NTPDase3, which hydrolyze extracellular nucleotide tri- and/or di-phosphates. In ATP bioluminescence assays using carotid bodies, the degradation of extracellular ATP was attenuated in the presence of the selective NTPDases inhibitor ARL67156, suggesting ATP-degrading activity by NTPDases in the tissue. These results suggest that ATP released from type I cells is degraded into ADP and adenosine 5′-monophosphate by NTPDases expressed in type II cells, and that ADP modulates type I cells via P2Y12 purinoceptors.

Preliminary Study on Microbial Deterioration Control and Effectiveness Evaluation in the Neolithic Prehistoric Archaeological Site of Dadiwan, Northwest China

Microbial deterioration as one of the widespread problems in archaeological site museums significantly affects their safety and exhibits. This paper systemically investigated the environments and conditions of microbial outbreaks in the Dadiwan No. F901 site museum, which is a representative archaeological site of prehistoric Yangshao culture. The morphology and harmful characteristics of the outbreak microorganisms were analyzed by microscopic techniques. The ultraviolet resistance of harmful microorganisms was also studied. Combining these findings with the original facilities of the site museum, a scientific and reasonable project was proposed to control and prevent the activity of harmful microorganisms. In addition, a 1% OIT/DCOIT biocide concentration was applied to inhibit microorganism-caused deterioration, in combination with mechanical removal based on laboratory tests and screening in situ. The effectiveness of microbial control was assessed using a portable microscope, ATP bioluminescence assay, and color difference detection. As a long-lasting preventive measure for microbial deterioration, an ultraviolet sterilization system can efficaciously prevent the re-outbreak of microbial deterioration to form a relatively stable dynamic balance for the surroundings of the site. This study is a resultful exploration in terms of microbial control and plays an important role in the sustainable protection of archaeological site museums.

Amino Acid and Glucose Fermentation Maintain ATP Content in Mouse and Human Malignant Glioma Cells.

Energy is necessary for tumor cell viability and growth. Aerobic glucose-driven lactic acid fermentation is a common metabolic phenotype seen in most cancers including malignant gliomas. This metabolic phenotype is linked to abnormalities in mitochondrial structure and function. A luciferin-luciferase bioluminescence ATP assay was used to measure the influence of amino acids, glucose, and oxygen on ATP content and viability in mouse (VM-M3 and CT-2A) and human (U-87MG) glioma cells that differed in cell biology, genetic background, and species origin. Oxygen consumption was measured using the Resipher system. Extracellular lactate and succinate were measured as end products of the glycolysis and glutaminolysis pathways, respectively. The results showed that: (1) glutamine was a source of ATP content irrespective of oxygen. No other amino acid could replace glutamine in sustaining ATP content and viability; (2) ATP content persisted in the absence of glucose and under hypoxia, ruling out substantial contribution through either glycolysis or oxidative phosphorylation (OxPhos) under these conditions; (3) Mitochondrial complex IV inhibition showed that oxygen consumption was not an accurate measure for ATP production through OxPhos. The glutaminase inhibitor, 6-diazo-5-oxo-L-norleucine (DON), reduced ATP content and succinate export in cells grown in glutamine. The data suggests that mitochondrial substrate level phosphorylation in the glutamine-driven glutaminolysis pathway contributes to ATP content in these glioma cells. A new model is presented highlighting the synergistic interaction between the high-throughput glycolysis and glutaminolysis pathways that drive malignant glioma growth and maintain ATP content through the aerobic fermentation of both glucose and glutamine.

Comparison of the ATP-bioluminescence assay and the standard colony counting method for evaluation BCG products

Comparison of the ATP-bioluminescence assay and the standard colony counting method for evaluation BCG products

Treatment of cardiomyocytes isolated from left ventricular myocardium of dogs with chronic heart failure with ALDA-1, an activator of aldehyde dehydrogenase-2, improves mitochondrial function

Abstract Background Reactive oxygen species (ROS) generated by dysfunctional mitochondria (MITO) of the failing heart plays an important role in lipid peroxidation of polyunsaturated fatty acids in cell and MITO membranes resulting in the production of aldehydes, such as 4-hydroxy-2-nonenal (4-HNE). MITO aldehyde dehydrogenase-2 (mALDH2) is a key enzyme for the detoxification of endogenous aldehyde substrates through NAD dependent oxidation. ALDH2 catalyzes toxic 4HNE to nontoxic 4HNA in cardiac cells. We previously showed that mALDH2 expression and activity are markedly reduced in MITO of failing LV cardiomyocytes (CM), a maladaptation that can lead to considerable accumulation of 4-HNE in MITO and ultimately to progressive LV systolic and diastolic dysfunction. In this study, we tested the hypothesis that exposure of failing cardiomyocytes to ALDA-1, an activator of mALDH2, improves MITO function. Methods Freshly isolated CM from LV myocardium of dogs (n = 7) with heart failure (HF, LV ejection fraction ≤35%) and of normal (NL) dogs (n = 4) were incubated for 1 hour in absence (baseline) and presence of Alda-1 at concentration of 10 µM. After incubation, ADP-stimulated respiration (ADPresp) and maximal rate of ATP synthesis (ATPmax) were measured using Strathklein respirometer and bioluminescence ATP assay kit. Results Treatment of isolated CM from HF dogs with 10 µM Alda-1 significantly increased ADPresp (277 ± 18 vs. 189 ± 12 nAtom O/min/mg, p&amp;lt;0.05) and ATPmax (30 ± 3 vs. 16 ± 3 nmoles/min/mg, p&amp;lt;0.05) compared to untreated CM of HF dogs. Treatment of CM from NL dogs with 10 µM Alda-1 had no effect on ADPresp (488 ± 18 vs. 484 ± 19 nAtom O/min/mg) or ATPmax (54 ± 3 vs. 52 ± 2 nmoles/min/mg) compared to untreated NL CM. Conclusions Treatment of CM from HF dogs with Alda-1 improved overall MITO function. Activation of mALDH2 represents a potential viable therapeutic target for treating HF.

Developing a Prolamin-Based Gel for Food Packaging: In-Vitro Assessment of Cytocompatibility.

Growing environmental concerns drive efforts to reduce packaging waste by adopting biodegradable polymers, coatings, and films. However, biodegradable materials used in packaging face challenges related to barrier properties, mechanical strength, and processing compatibility. A composite gel was developed using biodegradable compounds (prolamin, d-mannose, citric acid), as a coating to increase the oxygen barrier of food packaging materials. To improve gel stability and mechanical properties, the gels were physically cross-linked with particles synthesized from tetraethyl orthosilicate and tetramethyl orthosilicate precursors. Additionally, biocompatibility assessments were performed on human keratinocytes and fibroblasts, demonstrating the safety of the gels for consumer contact. The gel properties were characterized, including molecular structure, morphology, and topography. Biocompatibility of the gels was assessed using bioluminescent ATP assay to detect cell viability, lactate dehydrogenase assay to determine cell cytotoxicity, and a leukocyte stimulation test to detect inflammatory potential. A composite gel with strong oxygen barrier properties in low-humidity environments was prepared. Increasing the silane precursor to 50 wt% during gel preparation slowed degradation in water. The addition of citric acid decreased gel solubility. However, higher precursor amounts increased surface roughness, making the gel more brittle yet mechanically resistant. The increase of precursor in the gel also increased gel viscosity. Importantly, the gels showed no cytotoxicity on human keratinocytes or fibroblasts and had no inflammatory effects on leukocytes. This composite gel holds promise for oxygen barrier food packaging and is safe for consumer contact. Further research should focus on optimizing the stability of the oxygen barrier in humid environments and investigate the potential sensitizing effects of biodegradable materials on consumers.

Phage-Based Biosensing for Rapid and Specific Detection of Staphylococcus aureus.

Staphylococcus aureus (S. aureus) is a major foodborne pathogen. Rapid and specific detection is crucial for controlling staphylococcal food poisoning. This study reported a Staphylococcus phage named LSA2302 showing great potential for applications in the rapid detection of S. aureus. Its biological characteristics were identified, including growth properties and stability under different pH and temperature conditions. The genomic analysis revealed that the phage has no genes associated with pathogenicity or drug resistance. Then, the phage-functionalized magnetic beads (pMB), serving as a biological recognition element, were integrated with ATP bioluminescence assays to establish a biosensing method for S. aureus detection. The pMB enrichment brought high specificity and a tenfold increase in analytical sensitivity during detection. The whole detection process could be completed within 30 min, with a broad linear range of 1 × 104 to 1 × 108 CFU/mL and a limit of detection (LOD) of 2.43 × 103 CFU/mL. After a 2 h pre-cultivation, this method is capable of detecting bacteria as low as 1 CFU/mL. The recoveries of S. aureus in spiked skim milk and chicken samples were 81.07% to 99.17% and 86.98% to 104.62%, respectively. Our results indicated that phage-based biosensing can contribute to the detection of target pathogens in foods.

Manifestation of Heat-Induced Valuable Dietary Nucleotide Salvage in Food Prepared from Aged Fish in Fast Protein and Metabolites Liquid Chromatography, ATP-Bioluminescence Assay, and NMR Spectra

Dietary nucleotides and nucleosides, primarily inosine monophosphate (IMP) and the adenine nucleotide pool (ANP), are widely considered as essential nutrients responsible for multiple biological functions. Food prepared from meat and fish is the main source of these substances in the human diet, and it is extremely important to implement storage and processing techniques ensuring their maximum preservation and even accumulation during maturation or conditioning. In experiments with freshly refrigerated grass carp and defrosted Alaska pollock fillets it was discovered, initially using Fast Protein and Metabolites Liquid Chromatography and the ATP-bioluminescence test, and afterwards validated by NMR spectroscopy, that heat treatment identical to conventional culinary processing in aqueous or wet media at temperatures above 62 °C leads to nucleotide salvage (recovery) in aged fish. A significant increase in the concentration of IMP, and even an emergence of ANP substances, were reliably demonstrated in fish samples which had already partially or fully lost these components during prolonged storage due to the ATP breakdown metabolic reactions. Owing to this recovery, the nutritive value of ready-to-eat food can be higher than was initially evaluated in raw products before heat treatment: an effect that should certainly be considered in practical nutrition. Moreover, it is necessary to reconsider the widely acknowledged system of indices of freshness based on nucleotides and nucleosides elaborated a long time ago for raw meat and fish products.

ATP bioluminescence-based method for screening of multiclass antibiotic residues in animal derived foods

In present study, a new ATP bioluminescence assay with Geobacillus stearothermophilus ATCC12980 as an indicator bacterium was developed to screen multiclass antibiotic residues in animal derived foods. The development experiments includes the optimization on the usage of D-luciferin, bacterial ATP extraction condition, incubation condition of detection system and sample pre-treatment. Additionally, the validation experiments of sensitivity, specificity, accuracy, precision and stability were performed. This method can detect β-lactams (6), aminoglycosides (4), tetracyclines (4), sulfonamides (4), macrolides (4), lincosamides (1) and quinolones (3) in muscle and kidney of swine at or close to maximum residue limits (MRLs) established by European Union (EU). The false-positive rate was 0%, and the false negative rates were < 5%. The recovery rate of antibacterial drugs were between 60% and 120%, and the daily coefficients of variation were less than 15%, which meets the criteria in Commission Decision 2002/657/EC. The shelf life of the detection system was more than 5 months. Accordingly, the proposed ATP bioluminescence assay shows great potential for rapid, sensitive and high throughput detection of multiple antibiotic residues in animal foods, which can provide a preliminary screening informations of antibiotic residues for the following chemical confirmation experiments and expedite disposal decisions of unqualified food for enhanced food safety inspection.

Key Elements Affecting the Library’s CFU Concentration in Taiwan

Public libraries are popular gathering places, so understanding the factors that contribute to colony-forming unit (CFU) concentrations and how to minimize them is essential. This study aimed to investigate the factors that affect CFU concentrations in a public library, using air sampling (Bioluminescent ATP-assay) and statistical analysis software (SPSS) to collect and analyze data. The findings indicated that the CFU concentration in the library was significantly influenced by the air quality surrounding the building, the number of library visitors, and the hygiene and health of both visitors and employees. Additionally, indoor temperature and humidity were found to be key factors affecting CFU concentration. These findings suggest the need for better ventilation and air filtration systems, as well as regular cleaning and disinfection in public libraries. Furthermore, research is recommended to investigate other potential factors that may impact indoor air quality in public spaces.

Guilu Erxian Jiao enhances protein synthesis, glucose homeostasis, mitochondrial biogenesis and slow-twitch fibers in the skeletal muscle.

Guilu Erxian Jiao (GEJ) is a commonly used nutritional supplement due to its rich content of amino acids. It is also a traditional herbal medicine for improving degenerative joint. This study aimed to investigate the effect and mechanism of GEJ water extract (GEJ-WE) on skeletal muscle in C2C12 myotubes and C57BL/6J mice. Analysis of GEJ-WE were performed by high-performance liquid chromatography fingerprinting with chemical standards. Protein expression, mRNA level, glycogen content, mitochondria activity and ATP level were evaluated by western blots, real-time PCR, PAS staining, MTT and ATP bioluminescence assay, respectively. Skeletal muscle strength was evaluated by grip strength. Skeletal muscle volume, mass and fiber types were evaluated by micro computed tomography, histological analysis and immunofluorescence staining, respectively. Motor function was evaluated by rotarod performance and locomotor activity. In C2C12 myotubes, GEJ-WE significantly enhanced myogenic differentiation and myotube growth, protein synthesis signaling IGF-1/IGF-1R/IRS-1/Akt, Glut4 translocation, glycogen content, mitochondrial biogenesis signaling PGC-1α/NRF1/TFAM, mitochondrial activity and ATP production. However, IGF-1R antagonist AG1024 and PI3K inhibitor wortmannin reduced GEJ-WE-induced protein expression of MyHC, p-Akt, p-mTOR and p-GSK-3β, Glut4 translocation and glycogen content. In C57BL/6J mice, GEJ-WE not only upregulated protein synthesis and mitochondrial biogenesis signaling, but it also increased muscle volume, relative muscle weight, cross-sectional area of myofibers, glycogen content and transition of fast-to-slow type fibers of skeletal muscles. Moreover, GEJ-WE enhanced grip strength and motor activity of mice. In conclusion, the upregulation of protein synthesis, myogenic differentiation, glucose homeostasis, mitochondrial biogenesis and slow-twitch fibers contributes to the mechanisms of GEJ-WE on the enhancement of skeletal muscle mass and motor function.

Colistin potentiation in multidrug-resistant Acinetobacter baumannii by a non-cytotoxic guanidine derivative of silver.

A novel nano-formulation (NF) that sensitizes Acinetobacter baumannii (AB) to otherwise ineffective colistin is described in the present study. Infections due to multidrug resistant (MDR) AB represent a major therapeutic challenge, especially in situations of pre-existing colistin resistance (colR). Subsequently, boosting the effectiveness of colistin would be a better alternative tactic to treat AB infections rather than discovering a new class of antibiotics. We have previously demonstrated an NF comprising self-assembled guanidinium and ionic silver nanoparticles [AD-L@Ag(0)] to have anti-biofilm and bactericidal activity. We report NF AD-L@Ag(0) for the very first time for the potentiation of colistin in Gram-negative colistin-resistant bacteria. Our results implied that a combination of clinically relevant concentrations of colistin and AD-L@Ag(0) significantly decreased colistin-resistant AB bacterial growth and viability, which otherwise was elevated in the presence of only colistin. In this study, we have described various combinations of minimum inhibitory concentration (MIC) of colistin (MICcol, 1/2 MICcol, and 1/4 MICcol) and that of AD-L@Ag(0) [MICAD-L@Ag(0), 1/2 MICAD-L@Ag(0), and 1/4 MICAD-L@Ag(0)] and tested them against MDR AB culture. The results (in broth as well as in solid media) signified that AD-L@Ag(0) was able to potentiate the anti-microbial activity of colistin at sub-MIC concentrations. Furthermore, the viability and metabolic activity of bacterial cells were also measured by CTC fluorescence assay and ATP bioluminescence assay. The results of these assays were in perfect concordance with the scores of cultures (colony forming unit and culture turbidity). In addition, quantitative real-time PCR (qRT-PCR) was performed to unveil the expression of selected genes, DNAgyrA, DNAgyrB, and dac. These genes introduce negative supercoiling in the DNA, and hence are important for basic cellular processes. These genes, due to mutation, modified the Lipid A of bacteria, further resisting the uptake of colistin. Therefore, the expression of these genes was upregulated when AB was treated with only colistin, substantiating that AB is resistant to colistin, whereas the combinations of MICcol + MICAD-L@Ag(0) downregulated the expression of these genes, implying that the developed formulation can potentiate the efficiency of colistin. In conclusion, AD-L@Ag(0) can potentiate the proficiency of colistin, further enhancing colistin-mediated death of AB by putatively disrupting the outer membrane (OM) and facilitating bacterial death.

Investigation of detection level by ATP bioluminescence assay for fire blight pathogen &lt;i&gt;Erwinia amylovora&lt;/i&gt; monitoring in a laboratory environment

Investigation of detection level by ATP bioluminescence assay for fire blight pathogen &lt;i&gt;Erwinia amylovora&lt;/i&gt; monitoring in a laboratory environment