Pump Activity Research Articles

Elucidating the augmented resistance profile of Scedosporium/Lomentospora species to azoles in a cystic fibrosis mimic environment.

Scedosporium/Lomentospora species are ranked as the second most frequently isolated filamentous fungi from cystic fibrosis (CF) patients. Previously, we demonstrated that the minimum inhibitory concentration (MIC) for voriconazole and posaconazole increased when performed on a mucin-containing synthetic CF sputum medium (SCFM) compared to the standard medium, RPMI-1640. In this study, we have expanded the MIC comparison to four additional azoles and investigated characteristics linked to azole resistance in Scedosporium apiospermum, Scedosporium minutisporum, Scedosporium aurantiacum and Lomentospora prolificans. MIC was assayed by CLSI protocol, efflux pump activity was assessed by rhodamine 6G and sterols were analysed by gas chromatography-mass spectrometry (GC-MS). Overall, MICs for fluconazole, itraconazole, voriconazole, posaconazole, miconazole and ketoconazole increased by least 2-fold when susceptibility tests were performed using SCFM compared to RPMI. The activity of efflux pumps was similar in both media; however, in RPMI, but not in SCFM, the activity was induced by voriconazole and fluconazole. Additionally, MICs for those antifungals decreased more noticeably in SCFM than in RPMI in the presence of the efflux pump inhibitor PaβN. The SCFM-grown cells presented fewer sterols in their composition, and consequently higher membrane fluidity, than RPMI-grown cells. GC-MS analysis demonstrated a remodulation in the sterol profile in SCFM- compared to RPMI-grown cells. Accordingly, when the MIC assay was performed in the presence of the membrane stressor NaCl (3%), the susceptibility to voriconazole and fluconazole increased more in SCFM- than RPMI-grown cells. Scedosporium/Lomentospora species undergo cellular adaptations in SCFM that favours their growth in face of the challenges imposed by azole antifungals.

Quantitative MODS-Wayne assay for rapid detection of pyrazinamide resistance in Mycobacterium tuberculosis from sputum samples.

PZA susceptibility testing continues to be a challenge, particularly in countries with high TB incidence. In response to this pressing need, we have developed a quantitative MODS-Wayne (MODS-WQ) assay. This approach offers a direct and cost-effective solution representing a significant advancement in TB diagnostics, particularly benefiting resource-limited laboratories, primarily in developing regions. The MODS-WQ assay stands out for its ability to quantify pyrazinoic acid (POA) production, as a reliable indicator of PZA resistance. Unlike traditional qualitative assays, MODS-WQ eliminates the inherent subjectivity in interpretation, providing more accurate and actionable results. Moreover, the MODS-WQ approach accounts for critical factors influencing PZA resistance, including enzymatic efficiency and efflux pump activity. By integrating these factors into the detection process, our methodology offers a comprehensive understanding of PZA resistance levels, enabling tailored treatment strategies for patients.

Biofilm formation and polar lipid biosynthesis in Mycobacterium abscessus are inhibited by naphthylmethylpiperazine.

Mycobacterium abscessus is a biofilm-forming, non-tuberculous mycobacterium that is highly resistant to antibiotics. Bacterial efflux pumps contribute to biofilm formation, export of biofilm-associated lipids and antibiotic tolerance. The Resistance Nodulation Cell Division (RND) and ATP-Binding Cassette (ABC) families of efflux pumps export lipids to the mycobacterial cell surface. 1-(1-naphthyl methyl)-piperazine (NMP) is a chemosensitizer that causes membrane destabilization and is an inhibitor of RND efflux pumps. The effects of NMP on biofilm formation and lipid metabolism in M. abscessus biofilms have not been investigated. Plumbagin (PLU) is an inhibitor of ABC efflux pumps that has not been studied for its effects on antibiotic tolerance in M. abscessus biofilms. In this study, we report that the efflux pump inhibitors NMP and PLU inhibit biofilm formation by 50% at sub-MIC levels. We show that NMP inhibits the incorporation of the radiolabeled long-chain fatty acid 14C-palmitate into glycopeptidolipids in cell surface lipids of log-phase M. abscessus. NMP also inhibits the utilization of the radiolabel in the biosynthesis of phosphatidylethanolamine in the cell surface and cellular lipids of M. abscessus cells in log-phase and in biofilms. Incorporation of the radiolabel into cardiolipin in the cellular lipids of M. abscessus biofilms was inhibited by NMP. The incorporation of 14C-acetate into cell surface phosphatidylethanolamine in log-phase and biofilm cells was also inhibited by NMP. Triacylglycerol biosynthesis using 14C-palmitate and 14C-acetate in cellular lipids of log-phase and biofilm cells was increased several folds by NMP. Efflux pump activity in M. abscessus cells was inhibited by 97% and 68% by NMP and PLU respectively. NMP and PLU caused 5-fold decreases in the minimum inhibitory concentrations of ciprofloxacin and clarithromycin against M. abscessus. Our results demonstrate that NMP and PLU affect important physiological processes in M. abscessus associated with its pathogenesis.

Role of membrane H+ transport and plasmalemma excitability in pattern formation, long-distance transport and photosynthesis of Characean algae

Illuminated giant cells of Characeae comprise alternating areas with H+ pump activity and zones with high conductivity for H+/OH–, which create counter-directed H+ flows between the medium and the cytoplasm. In areas where H+ enters the cell, the pH on the surface (pHo) increases to pH 10, while the cytoplasmic pH (pHc) decreases. The lack of the permeant substrate of photosynthesis (CO2) and the acidic pHc shift in the region of external alkaline zones redirect electron transport in chloroplasts from CO2-dependent (assimilatory) pathway to O2 reduction. This electron transport route is associated with an increase in thylakoid membrane ΔpH and an enhanced nonphotochemical quenching (NPQ) of chlorophyll excitations, which underlies strict coordination between nonuniform distributions of pHo and photosynthetic activity in resting cells. When the action potential (AP) is generated, the longitudinal pH profile is temporarily smoothed out, while the heterogeneity of the distribution of NPQ and PSII photochemical activity (YII) sharply increases. The damping of the pHo profile is due to the suppression of the H+ pump and passive H+ conductance under the influence of an almost 100-fold increase in the cytoplasmic of Ca2+ level ([Ca2+]c) during AP. The increase in [Ca2+]c stimulates photoreduction of O2 in chloroplasts under external alkaline zones and, at the same time, arrests the cytoplasmic streaming, which causes the accumulation of excess amounts of H2O2 in the cytoplasm in areas of intense production of this metabolite, with a weak effect on areas of CO2 assimilation. These changes enhance the nonuniform distribution of cell photosynthesis and account for the long-term oscillations of chlorophyll fluorescence Fm' and the quantum efficiency of linear electron flow in microscopic cell areas after the AP generation.

Metabolism of nitrogen and sulfamethoxazole, performance recovery mechanism, and antibiotic resistance genes (ARGs) behaviors in membrane aerated biofilm reactor (MABR)

The limited information on micro-mechanism of sulfamethoxazole (SMX) metabolism and antibiotics resistance genes (ARGs) behaviors within membrane-aerated biofilm reactor (MABR) blocked the application of MABR in treating antibiotics containing wastewater. In this work, SMX metabolism and the changes of ARGs were investigated in MABR during 0.0-0.7 mg/L SMX. The system achieved a 92.3% SMX removal at 0.7 mg/LSMX, leading to the formation of 17 lower toxic metabolites, which indicated the robust SMX biodegradation capability of MABR. The metabolic pathways of SMX in MABR were first elucidated, revealing key processes including isoxazole ring opening (primarily in anaerobic zones), aniline hydroxylation, and cleavage of the S-N bond in the sulfonyl group. Microbial analysis revealed the involvement of multiple microbes in SMX biodegradation, and among these, sulfate-reducing bacteria (SRB; Desulfovibrio, Desulforhopalus, Desulfomicrobium) facilitated the isoxazole ring opening in the anaerobic zone, while Rhizobium and Chryseobacterium contributed to ipso-hydroxylation and cleavage of the -C-S-N bond in SMX, respectively. Furthermore, the slower accumulation of ARGs under SMX conditions was primarily attributed to the enhanced efflux pump activity, antibiotic target replacement mechanisms, and secretion of extracellular polymeric substances (EPS), suggesting the potential of MABR to inhibit ARG proliferation. Our research filled the knowledge gap in the micro-mechanism of SMX metabolism and ARGs fate within MABR processing SMX-containing wastewater, laying a theoretical foundation for the broader application of MABR in practical engineering.

Quantifying bacterial efflux within subcellular domains of Pseudomonas aeruginosa.

Molecular efflux is a mechanism through which bacteria actively expel undesirable substances. This is a crucial line of defense against toxic chemicals in harsh environments. Understanding how efflux works is critical for designing antimicrobial strategies. Though much is already known about efflux proteins, important details about the mechanisms of efflux (e.g., importance of specific subcellular domains and ejection rates) have yet to be experimentally quantified. Herein, we use the nonlinear optical technique, second harmonic light scattering, to simultaneously measure the efflux rates from the periplasm and cytosol of a Gram-negative bacterium. The influence of efflux on the uptake kinetics of a mild antimicrobial, malachite green (MG), by Pseudomonas aeruginosa was quantified. It is observed that efflux primarily occurs from the periplasm and is two orders of magnitude faster than from the cytosol. Efflux pumps activate to maintain MG concentrations in the periplasm below 1 µM, while efflux from the cytosol maintains MG concentration below 0.1 µM. Efflux pumps are shown to saturate when exogenous MG concentrations are greater than 25 µM, while the cytosol efflux function saturates at >15 µM. Finally, efflux pumps can simultaneously eject different compounds, as proven by experiments with both MG and hexane, a known effluxable compound.IMPORTANCEMolecular efflux pumps are a crucial defense mechanism that protects bacteria from an otherwise unchecked influx of toxic molecules present in the extracellular environment. The efflux functions constitute a significant hindrance to antimicrobial efficacy. While much is now known regarding the structure of these channels, knowledge of the influence of efflux in individual subcellular domains and the associated ejection rates is still lacking. Using the nonlinear optical technique, second-harmonic light scattering, we have measured the threshold concentrations for pump activation, saturation concentrations, and efflux rates from both the periplasm and cytosol in living Gram-negative bacteria. The quantified efflux data in the different subcellular compartments not only provide a clear mechanistic understanding but also are critical for developing antimicrobial strategies.

Antibiotic Resistance and Presence of Persister Cells in the Biofilm-like Environments in Streptococcus agalactiae

Objectives: This study investigated antibiotic resistance and presence of persister cells in Streptococcus agalactiae strains belonging to capsular types Ia/ST-103, III/ST-17, and V/ST-26 in biofilm-like environments. Results: S. agalactiae strains were susceptible to penicillin, clindamycin, and erythromycin. Resistance genes were associated with tetM (80%), tetO (20%), ermB (80%), and linB (40%). Persister cells were detected in bacterial strains exposed to high concentrations of penicillin, clindamycin, and erythromycin. S. agalactiae capsular type III/ST-17 exhibited the highest percentage of persister cells in response to penicillin and clindamycin, while type Ia/ST-103 presented the lowest percentages of persister cells for all antimicrobials tested. Additionally, persister cells were also detected at lower levels for erythromycin, regardless of capsular type or sequence type. Further, all S. agalactiae isolates presented efflux pump activity in ethidium bromide-refractory cell assays. LIVE/DEAD fluorescence microscopy confirmed the presence of >85% viable persister cells after antibiotic treatment. Conclusions: These findings suggest that persister cells play a key role in the persistence of S. agalactiae during antibiotic therapy, interfering with the treatment of invasive infections. Monitoring persister formation is crucial for developing strategies to combat recurrent infections caused by this pathogen.

Groundwater-surface water exchanges in an alluvial plain in southern France subjected to pumping: A coupled multitracer and modeling approach

Study regionThe study was conducted in an alluvial plain between the Rhône and the Ouvèze Rivers (in the southeast of France) extensively exploited for drinking water. The research area is characterized by significant groundwater-surface interactions influenced by groundwater pumping activities. Study focusThe aim of this study is to enhance the understanding of interactions between rivers and alluvial aquifers by combined multi-tracer and numerical modeling approaches. Over an 18-month period, groundwater temperature, piezometric levels, and river surface water levels were continuously monitored. Field campaigns focused on conductivity, stable isotopes of water, and radon-222 activity concentration in both groundwater and surface water. Radon-222 was used to quantify water exchanges between the river and the aquifer. A MODFLOW model, calibrated using piezometric data and PEST, was employed to simulate groundwater flow and reactive transport of radon-222 using MT3DMS. New hydrological insights for the regionThe study reveals that river water recharges the aquifer, with radon-222 data delineating this recharge zone. The methodology extended the interpretation of periodic groundwater temperature signals to isotopic signals, allowing the identification of dispersivity and Darcy's velocity. The Ouvèze River was found to contribute approximately 55 % of the pumping water supply, alongside the Rhône. These findings provide valuable insights for sustainable water resource management, demonstrating the relevance of using natural tracers in scenarios where artificial tracers are impractical.

Enhancing groundwater management with GRACE-based groundwater estimates from GLDAS-2.2: a case study of the Almonte-Marismas aquifer, Spain

AbstractThe Almonte-Marismas aquifer, southwestern Spain, is a critical ecohydrogeological system that features extensive groundwater monitoring. This study investigates the utility of gravity recovery and climate experiment (GRACE) satellite data, specifically obtained from the global land data assimilation system (GLDAS) version 2.2, for assessing groundwater storage variations in the Almonte-Marismas aquifer. The presented research emphasizes the practical application of readily available GLDAS products that do not require data preprocessing. The study validates the GLDAS-2.2-based ready-to-use groundwater storage (GWS) time series by correlating it with precipitation and piezometric information, highlighting its effectiveness in medium-scale aquifers. The results reveal a strong agreement between GLDAS-2.2-derived GWS anomalies and in-situ measurements, confirming GLDAS-2.2’s potential for assessing aquifer depletion. The study discusses the consistency of seasonal variations in groundwater levels and GLDAS-2.2 data, emphasizing their close alignment with precipitation and pumping activities. Importantly, the study introduces GLDAS-2.2-derived volumetric groundwater storage (VGWS) as a valuable calibration parameter for numerical groundwater flow models, enhancing their accuracy over time. Moreover, the analysis reveals disparities in annual recharge values between GLDAS-2.2-derived data and the soil-water mass balance. These variations suggest the importance of additional inputs to precipitation, possibly related to subsurface or lateral connections. Overall, this study contributes to the ongoing discourse on the practical applications of GLDAS-2.2-derived GWS data in groundwater management, offering insights into its effectiveness in diverse hydrogeological settings, particularly in areas that lack monitoring infrastructure.

Histidine-rich calcium-binding protein: a molecular integrator of cardiac excitation-contraction coupling.

During mammalian cardiomyocyte excitation-contraction coupling, Ca2+ influx through voltage-gated Ca2+ channels triggers Ca2+ release from the sarcoplasmic reticulum (SR) through ryanodine receptor channels. This Ca2+-induced Ca2+ release mechanism controls cardiomyocyte contraction and is exquisitely regulated by SR Ca2+ levels. The histidine-rich calcium-binding protein (HRC) and its aspartic acid-rich paralogue aspolin are high-capacity, low-affinity Ca2+-binding proteins. Aspolin also acts as a trimethylamine N-oxide demethylase. At low intraluminal Ca2+ concentrations, HRC binds to the SR Ca2+-ATPase 2, inhibiting its Ca2+-pumping activity. At high intraluminal Ca2+ levels, HRC interacts with triadin to reduce Ca2+ release through ryanodine receptor channels. This Review analyses the evolution of these Ca2+-regulatory proteins, to gain insights into their roles. It reveals that HRC homologues are present in chordates, annelid worms, molluscs, corals and sea anemones. In contrast, triadin appears to be a chordate innovation. Furthermore, HRC is evolving more rapidly than other cardiac excitation-contraction coupling proteins. This positive selection (or relaxed negative selection) occurs along most of the mammalian HRC protein sequence, with the exception being the C-terminal cysteine-rich region, which is undergoing negative selection. The histidine-rich region of HRC might be involved in pH sensing, as an adaptation to air-breathing, endothermic and terrestrial life. In addition, a cysteine-rich pattern within HRC and aspolin is also found in a wide range of iron-sulfur cluster proteins, suggesting roles in redox reactions and metal binding. The polyaspartic regions of aspolins are likely to underlie their trimethylamine N-oxide demethylase activity, which might be mimicked by the acidic regions of HRCs. These potential roles of HRCs and aspolins await experimental verification.

The roles of GLUT5 in cancer progression, metastasis, and drug resistance

Emerging evidence has suggested that high fructose intake, particularly from added sugars and processed foods, is associated with increased cancer risk and progression. The fructose intake is believed to be mediated by the abnormal expression of glucose transporter 5 (GLUT5), the specific fructose transporter in cancer cells. The GLUT5-regulated fructose metabolism has shown to greatly affect cancer progression, metastasis, and drug resistance. This review aims to synchronize the current knowledge to highlight the underlying mechanisms of those impacts and understand the therapeutic potential of GULT5. First, we review the fructose metabolism and its alteration in cancer cells by comparing with glucose metabolism. Subsequently, the key contributors or biological pathways involved in GLUT5-asosociated tumor growth, cancer metastasis, and drug resistance are discussed. The contributions of specific pathways, metabolites, and key enzymes from the fructose metabolism process are also covered, such as enhanced glycolysis for tumor growth, epithelial-mesenchymal transition and angiogenesis for cancer metastasis, and efflux pump expression and activation of survival pathways for cancer drug resistance. The detailed analysis of these mechanisms will allow further understanding of the therapeutic potential of GLUT 5-mediated fructose metabolism in cancer therapy. In particular, targeting GLUT 5 and its-associated processes in fructose metabolism may offer promising strategies for improving cancer treatment outcomes through dietary interventions, specific GLUT5 inhibitors, or in combination.

Characterization of Chlorhexidine Resistance Among Clinical Isolates of Coagulase-Negative Staphylococcus Species in a Tertiary Care Center, Chennai.

Background Coagulase-negativeStaphylococci(CoNS) are potential pathogens and are often associated with healthcare-associated infections (HAIs). Chlorhexidine (CHX) is the most widely used antiseptic to reduce colonization and infection by allStaphylococci, including CoNS. Resistance to CHXamong CoNS has been observed over the past few years, consequent to its widespread use. Phenotypic tolerance or reduced susceptibility to CHX is conferred by plasmid-mediatedqacgroup of genes, mainlyqacA/Bandsmr, which cause activation of efflux pumps over the bacterial cell wall. This study aims to characterize the phenotypic and genotypic resistance exhibited by CoNS species against CHX. Methods After ethical approval, 148 consecutive, non-repetitive isolates of clinically significant CoNS species of hospitalized patients, isolated from blood samples and exudative specimens, were included in the study. Speciation was performed by conventional biochemical identification and automated methods. Antimicrobial susceptibility testing was performed by disc diffusion technique and for vancomycin by minimum inhibitory concentration (MIC) determination, as per Clinical Laboratory Standards Institute (CLSI) M-100 2023 guidelines. Methicillin resistance was detected using a cefoxitin disc. MIC for CHX was performed by agar dilution method; reduced susceptibility was considered when MIC to CHX ≥4 µg/mL. The simplex polymerase chain reaction (PCR) was carried out with suitable controls to detectqacA/Bandsmr. Statistical analysis was conducted to determine the association ofqacA/Bandsmrgenes with MIC of CHXin the study isolates. Results Fifteen different species of CoNS were obtained from clinical samples. A high percentage of resistance was observed against various classes of antibiotics. Methicillin resistance was observed in 69.6% (103/148) of isolates. Of 148 CoNS, 52.7% (78/148) of isolates exhibited reduced susceptibility to CHX with an MIC ≥4 µg/mL. These isolates exhibited a higher percentage of methicillin resistance (75.6%, 59/78). By PCR, 34.5% (51/148) of isolates carried either or both genes. GeneqacA/Bwas solely detected in 27.02% (40/148) of isolates, of which 14 were CHX-tolerant and the remaining 26 were CHX-susceptible. Genesmrwas solely detected in 4.1% (6/148) of isolates comprising three isolates each in CHX-tolerant and susceptible categories.There were 3.4% (5/148) of isolates that harbored bothgenes, of which only one isolate was CHX-susceptible, while the other four were CHX-tolerant. A proportion of isolates that were phenotypically tolerant to CHX did not carry either or both genes. A significant statistical association was found between reduced susceptibility to CHX and the presence of antiseptic resistance genesin the study isolates (p-value=0.033942). Conclusion To our knowledge, this is the first study from South India to investigate CHX resistance among CoNS using phenotypic and genotypic methods. The rise of antiseptic resistance among CoNS is an emerging threat to current infection control practices. The presence ofqacA/Bandsmrgenes, especially in CHX susceptible isolates, is concerning since these resistance genes are located on transferable plasmids, and the isolates can develop resistance eventually upon exposure to CHX.

Advancing glider-based acoustic measurements of underwater-radiated ship noise.

Ocean gliders are versatile and efficient passive acoustic monitoring platforms in remote marine environments, but few studies have examined their potential to monitor ship underwater noise. This study investigates a Slocum glider's capability to assess ship noise compared to the ability of fixed observers. Trials were conducted in shallow coastal inlets and deep bays in Newfoundland, Canada, using a glider, hydrophone array, and single-moored system. The study focused on (1) the glider's self-noise signature, (2) range-depth-dependent propagation loss (PL) models, and (3) identifying the location of the vessel to the glider using glider acoustic measurements. The primary contributors to the glider's self-noise were the buoyancy pump and rudder. The pitch-motor noise coincided with the buoyancy pump activation and did not contribute to the glider self-noise in our experiments. PL models showed that seafloor bathymetry and sound speed profiles significantly impacted estimates compared to models assuming flat and range-independent profiles. The glider's performance in recording ship noise was superior to that of other platforms. Using its hydrophones, the glider could identify the bearing from the vessel, although a third hydrophone would improve reliability and provide range. The findings demonstrate that gliders can characterize noise and enhance our understanding of ocean sound sources.

Metformin and vitamin D combination therapy ameliorates type 2 diabetes mellitus-induced renal injury in male Wistar rats.

Diabetic kidney disease is a major microvascular diabetes mellitus (DM) complication clinically associated with a gradual renal function decline. Although metformin is a common drug for managing DM, however, monotherapy treatment with any antidiabetic drug will necessitate dosage increment since type 2 DM (T2DM) deteriorates over time due to the increasing pancreatic β-cell dysfunction and will eventually require a combination therapy approach with another antidiabetic medication. Vitamin D is a food supplement that has been proven to have antidiabetic and reno-protective activities. Hence, we explore the combination of vitamin D and metformin on T2DM-induced renal dysfunction. Thirty male Wistar rats were randomized into five (5) groups: control, diabetes untreated, diabetics treated with metformin, vitamin D, and vitamin D + metformin. Vitamin D and metformin significantly reversed DM-induced hyperglycemia, electrolyte imbalance, and dyslipidemia. Also, vitamin D and metformin reversed T2DM-induced increase in serum creatinine and urea and renal lactate, LDH, and oxido-inflammatory response. These observed alterations were accompanied by an increase in proton pump activities and modulation of Nrf2/Nf-κB and XO/UA signaling. This study revealed that vitamin D and/or metformin ameliorated T2DM-induced renal injury.

Resistance Mechanisms of Plant Pathogenic Fungi to Fungicide, Environmental Impacts of Fungicides, and Sustainable Solutions

The significant reduction in agricultural output and the decline in product quality are two of the most glaring negative impacts caused by plant pathogenic fungi (PPF). Furthermore, contaminated food or transit might introduce mycotoxins produced by PPF directly into the food chain. Eating food tainted with mycotoxin is extremely dangerous for both human and animal health. Using fungicides is the first choice to control PPF or their toxins in food. Fungicide resistance and its effects on the environment and public health are becoming more and more of a concern, despite the fact that chemical fungicides are used to limit PPF toxicity and control growth in crops. Fungicides induce target site alteration and efflux pump activation, and mutations in PPF result in resistance. As a result, global trends are shifting away from chemically manufactured pesticides and toward managing fungal plant diseases using various biocontrol techniques, tactics, and approaches. However, surveillance programs to monitor fungicide resistance and their environmental impact are much fewer compared to bacterial antibiotic resistance surveillance programs. In this review, we discuss the PPF that contributes to disease development in plants, the fungicides used against them, factors causing the spread of PPF and the emergence of new strains, the antifungal resistance mechanisms of PPF, health, the environmental impacts of fungicides, and the use of biocontrol agents (BCAs), antimicrobial peptides (AMPs), and nanotechnologies to control PPF as a safe and eco-friendly alternative to fungicides.

Investigating Skeletal Muscle Pump Function Using Thigh EMG, Thigh NIRS, And Stroke Volume Data In The FRAILMatics Physiological Cohort

Abstract Background The pioneering female physiologist Frances Hellebrandt first highlighted the significance of the skeletal muscle pump in circulation in the 1930s. However, the mechanisms underlying the muscle pump's function remain elusive, particularly in older individuals. It is thought that during muscle contraction, increased intramuscular pressure facilitates the expulsion of blood from the muscle's veins, aiding venous return. This contraction-induced pressure gradient assists in pushing blood towards the heart, increasing preload, and subsequently stroke volume (SV). In this study we investigated muscle pump activity using continuous measurements of thigh muscle activity, thigh total haemoglobin concentration (Hb), and cardiac SV. Methods 22 individuals (aged 70.8±5.3 years, 77.3% female) recruited from a Falls and Syncope Unit (St James’s Hospital, Dublin) underwent non-invasive measurements of Hb (NIRS PortaLite, Artinis Medical Systems, The Netherlands), electromyography (EMG) (Shimmer3, Research Ltd, Ireland), and SV (Finapres Nova, Finapres Medical Systems, The Netherlands) during a 10-second thigh squeeze test. Pre-processing involved data quality checks, alignment, and resampling. 60-second regions of interest were extracted, normalised, and processed using MATLAB (R2023a, TheMathWorks, Inc., Natick, MA, USA). Cross-correlation and Granger causality analyses were conducted to explore relationships between signals. Results Thigh Hb and EMG signals lagged 8.4 and 8.5 seconds behind SV, respectively. Granger Causality analysis revealed a significant causal relationship from both Hb and EMG to SV in 68% of participants at measured lags. Conclusion The observed ~8.5-second lag between thigh measurements and SV implies a plausible temporal relationship between muscle activation and venous return, further supported by significant causal relationships found in a substantial portion of participants using Granger causality analysis. These findings underscore the importance of continued research in this area, not only for advancing our understanding of physiological mechanisms but also for informing potential clinical applications aimed at optimising cardiovascular health and function.

Mechanism of synergistic action of colistin with resveratrol and baicalin against mcr-1-positive Escherichia coli

The rising incidence of colistin (COL) resistance poses a significant challenge, undermining the therapeutic efficacy of COL against life-threatening bacterial infections. Therefore, the urgent identification and development of new therapeutics are imperative. It has been proven that combinations of antibiotics and promising non-antibiotic agents could be a potential strategy to combat infections caused by MDR pathogens. Due to various antimicrobial properties, medicinal plants have attracted significant attention, which could be promising adjuvant. In this study, we investigated the synergistic effects of combining COL with resveratrol (RST) and baicalin (BAI) against mcr-1-positive Escherichia coli through antibiotic susceptibility testing, checkerboard method and time-killing assays. The mechanisms of combination treatment were analyzed using SEM, fluorometric assays and transcriptome analysis. The molecular docking assay was conducted to elucidate potential interactions between RST, BAI and the MCR-1 protein. Finally, we assessed the in vivo efficacy of combination against mcr-1-positive Escherichia coli. The results demonstrated that the combination of RST, BAI and COL showed significant synergistic activity both in vitro and in vivo. Further mechanistic study revealed that the combination could increase the membrane-damaging ability of COL, disrupt the homeostasis of proton motive force (PMF), inhibit the activity of efflux pumps and impair ATP supply. The molecular docking revealed that RST and BAI could bind to MCR-1 stably, indicating the combination of RST and BAI may be an effective MCR-1 inhibitor. Our findings demonstrated that the combination of RST and BAI might be potential COL adjuvant, providing an alternative approach to address mcr-1-positive Escherichia coli infections.

Influence of Intraocular Pressure on the Expression and Activity of Sodium-Potassium Pumps in the Corneal Endothelium.

The corneal endothelium is responsible for pumping fluid out of the stroma in order to maintain corneal transparency, which depends in part on the expression and activity of sodium-potassium pumps. In this study, we evaluated how physiologic pressure and flow influence transcription, protein expression, and activity of Na+/K+-ATPase. Native and engineered corneal endothelia were cultured in a bioreactor in the presence of pressure and flow (hydrodynamic culture condition) or in a Petri dish (static culture condition). Transcription of ATP1A1 was assessed using qPCR, the expression of the α1 subunit of Na+/K+-ATPase was measured using Western blots and ELISA assays, and Na+/K+-ATPase activity was evaluated using an ATPase assay in the presence of ouabain. Results show that physiologic pressure and flow increase the transcription and the protein expression of Na+/K+-ATPase α1 in engineered corneal endothelia, while they remain stable in native corneal endothelia. Interestingly, the activity of Na+/K+-ATPase was increased in the presence of physiologic pressure and flow in both native and engineered corneal endothelia. These findings highlight the role of the in vivo environment on the functionality of the corneal endothelium.

Bismuth-based drugs sensitize Pseudomonas aeruginosa to multiple antibiotics by disrupting iron homeostasis.

Pseudomonas aeruginosa infections are difficult to treat due to rapid development of antibiotic drug resistance. The synergistic combination of already-in-use drugs is an alternative to developing new antibiotics to combat antibiotic-resistant bacteria. Here we demonstrate that bismuth-based drugs (bismuth subsalicylate, colloidal bismuth subcitrate) in combination with different classes of antibiotics (tetracyclines, macrolides, quinolones, rifamycins and so on) can eliminate multidrug-resistant P. aeruginosa and do not induce development of antibiotic resistance. Bismuth disrupts iron homeostasis by binding to P. aeruginosa siderophores. Inside cells, bismuth inhibits the electron transport chain, dissipates the proton motive force and impairs efflux pump activity by disrupting iron-sulfur cluster-containing enzymes, including respiration complexes. As a result, bismuth facilitates antibiotic accumulation inside bacteria, enhancing their efficacy. The combination therapy shows potent antibacterial efficacy and low toxicity in an ex vivo bacteraemia model and increases the survival rate of mice in in vivo mouse lung-infection models. Our findings highlight the potential of bismuth-based drugs to be repurposed to combat P. aeruginosa infections in combination with clinically used antibiotics.

Multistability of bursting rhythms in a half-center oscillator and the protective effects of synaptic inhibition.

For animals to meet environmental challenges, the activity patterns of specialized oscillatory neural circuits, central pattern generators (CPGs), controlling rhythmic movements like breathing and locomotion, are adjusted by neuromodulation. As a representative example, the leech heartbeat is controlled by a CPG driven by two pairs of mutually inhibitory interneurons, heart interneuron (HN) half-center oscillators (HCO). Experiments and modeling indicate that neuromodulation of HCO navigates this CPG between dysfunctional regimes by employing a co-regulating inverted relation; reducing Na+/K+ pump current and increasing hyperpolarization-activated (h-) current. Simply reducing pump activity or increasing h-current leads to either seizure-like bursting or an asymmetric bursting dysfunctional regime, respectively. Here, we demonstrate through modeling that, alongside this coregulation path, a new bursting regime emerges. Both regimes fulfill the criteria for functional bursting activity. Although the cycle periods and burst durations of these patterns are roughly the same, the new one exhibits an intra-burst spike frequency that is twice as high as the other. This finding suggests that neuromodulation could introduce additional functional regimes with higher spike frequency, and thus more effective synaptic transmission to motor neurons. We found that this new regime co-exists with the original bursting. The HCO can be switched between them by a short pulse of excitatory or inhibitory conductance. In this domain of coexisting functional patterns, an isolated cell model exhibits only one regime, a severely dysfunctional plateau-containing, seizure-like activity. This aligns with widely reported notion that deficiency of inhibition can cause seizures and other dysfunctional neural activities. We show that along the coregulation path of neuromodulation, the high excitability of the single HNs induced by myomodulin is harnessed by mutually inhibitory synaptic interactions of the HCO into the functional bursting pattern.