Rapid Adenosine Research Articles

Transient Adenosine Modulates Serotonin Release Indirectly in the Dorsal Raphe Nuclei.

Rapid adenosine transiently regulates dopamine and glutamate via A1 receptors, but other neurotransmitters, such as serotonin, have not been studied. In this study, we examined the rapid modulatory effect of adenosine on serotonin release in the dorsal raphe nuclei (DRN) of mouse brain slices by using fast-scan cyclic voltammetry. To mimic adenosine release during damage, a rapid microinjection of adenosine at 50 pmol was applied before electrical stimulation of serotonin release. Transient adenosine significantly reduced electrically evoked serotonin release in the first 20 s after application, but serotonin release recovered to baseline as adenosine was cleared from the slice. The continuous perfusion of adenosine did not change the evoked serotonin release. Surprisingly, the modulatory effects of adenosine were not regulated by A1 receptors as adenosine still inhibited serotonin release in A1KO mice and also after perfusion of an A1 antagonist (8-cyclopentyl-1,3-dipropyl xanthine). The inhibition was also not regulated by A3 receptors as perfusion of the A3 antagonist (MRS 1220) in A1KO brain slices did not eliminate the inhibitory effects of transient adenosine. In addition, adenosine also inhibited serotonin release in A2AKO mice, showing that A2A did not modulate serotonin. However, perfusion of a selective 5HT1A autoreceptor antagonist drug [(S)-WAY 100135 dihydrochloride] abolished the inhibitory effect of transient adenosine on serotonin release. Thus, the transient neuromodulatory effect of adenosine on DRN serotonin release is regulated by serotonin autoreceptors and not by adenosine receptors. Rapid, transient adenosine modulation of neurotransmitters such as serotonin may have important implications for diseases such as depression and brain injury.

Xanthine oxidase inhibitors treatment or discontinuation effects on mortality: evidence of xanthine oxidase inhibitors withdrawal syndrome.

Objectives: This study investigates the impact of xanthine oxidase inhibitors (XOI) on mortality in patients with cardiovascular diseases. XOI withdrawal has been reported to increased mortality risk due to rapid adenosine triphosphate (ATP) deficiency. This study aims to determine whether XOI treatment reduces mortality and whether XOI withdrawal increases mortality. Methods: This is a real-world database study using the Japanese Registry of All Cardiac and Vascular Diseases (J-ROAD). We analyzed 1,648,891 hospitalized patients aged 20-90 with acute coronary syndrome or heart failure. In the first study, mortality rates were compared between patients without urate-lowering agents (n = 1,292,486) and those with XOI agents (n = 315,388, excluding 41,017 on other urate-lowering agents). In the second study, mortality rates were compared between the XOI continuous medication group (n = 226,261) and the XOI withdrawal group (n = 89,127). Results: After multiple adjustments, XOI treatment group showed significantly lower mortality compared with that without any urate-lowering agent (odds ratio (OR), 0.576, 95% confidence interval (CI), 0.567-0.587, p < .001). In the sub-analysis, the group with allopurinol (OR, 0.578; 95% CI, 0.557-0.600), febuxostat (OR, 0.610; 95% CI, 0.599-0.622), and topiroxostat (HR, 0.545; 95% CI, 0.473-0.628) showed lower OR of mortality compared with that without any urate-lowering agent. XOI withdrawal group led to significantly higher death rates compared to XOI continuous group (19.8% vs. 0.03%; p < .001). Conclusion: XOI treatment for patients with cardiovascular diseases is associated with reduced mortality. Conversely, XOI withdrawal is linked to elevated mortality risk. This emphasizes the importance of both prescribing and discontinuing XOI carefully to optimize patient outcomes.

Adenosine triphosphate (ATP) sampling algorithm for monitoring the cleanliness of surgical instruments.

Timely detection of cleaning failure is critical for quality assurance within Sterilising Service Units (SSUs). Rapid Adenosine Triphosphate (ATP) testing provides a real time and quantitative indication of cellular contaminants, when used to measure surface or device cleanliness. The aim of this study was to investigate the use of an ATP algorithm and to whether it could be used as a routine quality assurance step, to monitor surgical instruments cleanliness in SSUs prior to sterilisation. Cleanliness monitoring using rapid ATP testing was undertaken in the SSUs of four hospitals located in the western (Amazonia) region of Brazil. ATP testing was conducted (Clean Trace, 3M) on 163 surgical instruments, following manual cleaning. A sampling algorithm using a duplicate swab approach was applied to indicate surgical instruments as (i) very clean, (ii) clean, (iii) equivocal or (iv) fail, based around a 'clean' cut-off of 250 Relative Light Units (RLU) and a 'very clean' <100 RLU. The four cleanliness categories were significantly differentiated (P≤0.001). The worst performing locations (hospitals A & C) had failure rates of 39.2% and 32.4%, respectively, and were distinctly different from hospitals B & D (P≤0.001). The best performing hospitals (B & D) had failure rates of 7.7% and 2.8%, respectively. The ATP testing algorithm provides a simple to use method within SSUs. The measurements are in real time, quantitative and useful for risk-based quality assurance monitoring, and the tool can be used for staff training. The four-tiered approach to the grading of surgical instrument cleanliness provides a nuanced approach for continuous quality improvement within SSU than does a simple pass/fail methodology.

Flexible needle-type Microbiosensor for real-time monitoring traditional acupuncture-mediated adenosine release In vivo

Rapid adenosine (ADO) signaling, on the time frame of seconds, regulates physiological and pathological processes, including the therapeutic efficacy of acupuncture. Nevertheless, standard monitoring strategies are limited by poor temporal resolution. Herein, an implantable needle-type microsensor capable of monitoring ADO release in vivo in response to acupuncture in real time has been developed. Electrocatalytic Prussian Blue nanoparticles, an immobilized multienzyme system, and a permselective poly-o-phenylenediamine-based membrane were used for the sequential modification of the sensing region of the electrode. The resultant sensor can perform amperometric measurements of ADO levels in response to a very low level of applied potential (−0.05 V vs Ag/AgCl). This microsensor also functioned across a broad linear range (0–50 μM) and exhibited good sensitivity (1.1 nA/μM) with a rapid response time of under 5 s. Importantly, the sensor also exhibited good reproducibility and high selectivity. For in vivo animal studies, the microsensor was employed for the continuous assessment of instantaneous ADO release at the ST36 (Zusanli) acupoint when this acupoint was subjected to twirling-rotating acupuncture manipulation. Benefiting from superior sensor in vivo performance and stability, the positive correlation between the variability in acupuncture-induced ADO release and the stimulus intensity levels that affect the clinical benefit can be demonstrated for the first time. Overall, these results highlight a powerful approach to analyzing the in vivo physiological effects of acupuncture, expanding application realm of micro-nano sensor technology on a fast time scale.

Does an Extraoral Suction Device Reduce Aerosol Generation and Prevent Droplet Exposure to the Examiner during Esophagogastroduodenoscopy?

Esophagogastroduodenoscopy (EGD) is an aerosol-generating procedure. A major challenge in the COVID-19 era is how to prevent the spread of aerosols and droplets in endoscopic units. We evaluated the effectiveness of an extraoral suction device in preventing indoor aerosol diffusion and droplet exposure for examiners. The study involved 61 patients who underwent EGD at our institution from 1 February to 31 March 2022. To determine whether aerosol spread increases before or after EGD examination with an extraoral suction device located in front of the patient's mouth, aerosols of 0.3, 0.5, 1, 3, 5, and 10 μm were measured with a handheld particle counter. The degree of contamination of the plastic gowns on the examiners was assessed using the rapid adenosine triphosphate test. The extraoral suction device significantly reduced the diffusion of large particles (3, 5, and 10 μm) after finishing the EGD examination. However, the diffusion of small particles (0.3 and 0.5 μm) was significantly increased. This extraoral suction device was effective in reducing large particle diffusion during EGD examination but was limited for minimizing small particle diffusion or droplet exposure to the examiner.

Effects of flash-boil shucking on biochemical, sensorial and ultrastructural properties of Yesso Scallop (Mizuhopecten yessoensis) adductor muscle during refrigeration

Abstract Scallop adductor muscle has great popularity for raw consumption in Japan, but conventionally shucking live scallops with high vitality by knife requires skill and is time-consuming. Flash-boil shucking is a convenient shucking method in sushi restaurants for live scallops. However, the effects of this treatment on the quality of its main edible part, the adductor muscle, are not clear. This study was conducted to investigate the impact of flash-boil shucking on the quality of raw-edible scallop adductor muscle, focusing on flavor-enhancing components and ultrastructure. Thermal treatment could contribute to the formation of flavor-enhancing nucleotides in the external layer of the adductor muscle. Adenosine monophosphate content increased with the boiling time, which significantly increased to 1.47±0.42 µmol/g in the 30 s-boiled group compared with 0.38±0.13 µmol/g in the control group (p&amp;lt;0.05). The rapid adenosine monophosphate accumulation after boiling resulted in a faster accumulation of inosine and hypoxanthine during subsequent refrigeration, which could bring bitterness to aquatic products. Flash-boil shucking had no discernible effect on the proportion of free amino acids relevant to the flavor of scallop adductor muscle, despite the possibility of a reduction in total free amino acids. The external layer of the scallop adductor muscle could be hardened after flash-boil shucking. At the ultrastructural level, muscle fibers tend to aggregate transversely and contract longitudinally depending on the heating time. Flash-boil shucking is ideal for consuming the raw-edible scallop adductor muscle promptly. Flash-boil shucking treatment is recommended for households, restaurants, and small factories for treating scallops with high vitality.

Regulation of the DEAH/RHA helicase Prp43 by the G-patch factor Pfa1

The DEAH/RHA helicase Prp43 remodels protein-RNA complexes during pre-messenger RNA (mRNA) splicing and ribosome biogenesis. The helicase activity and ATP turnover are intrinsically low and become activated by G-patch (gp) factors in the specific cellular context. The gp motif connects the helicase core to the flexible C-terminal domains, but it is unclear how this affects RecA domain movement during catalysis and the unwinding of RNA substrates. We developed single-molecule Förster Resonance Energy Transfer (smFRET)reporters to study RecA domain movements within Prp43 in real time. Without Pfa1(gp), the domains approach each other adopting predominantly a closed conformation. The addition of Pfa1(gp) induces an open state, which becomes even more prevalent during interaction with RNA. In the open state, Prp43 has reduced contacts with bound nucleotide and shows rapid adenosine diphosphate (ADP) release accelerating the transition from the weak (ADP) to the strong (apo) RNA binding state. Using smFRET labels on the RNA to probe substrate binding and unwinding, we demonstrate that Pfa1(gp) enables Prp43(ADP) to switch between RNA-bound and RNA-unbound states instead of dissociating from the RNA. ATP binding to the apo-enzyme induces the translocation along the RNA, generating the unwinding force required to melt proximal RNA structures. During ATP turnover, Pfa1(gp) stimulates alternating of the RecA domains between open and closed states. Consequently, the translocation becomes faster than dissociation from the substrate in the ADP state, allowing processive movement along the RNA. We provide a mechanistic model of DEAH/RHA helicase motility and reveal the principles of Prp43 regulation by G-patch proteins.

Pannexin1 channels regulate mechanically stimulated but not spontaneous adenosine release.

Fast-scan cyclic voltammetry (FSCV)is a rapid technique to measure neuromodulators, and using FSCV, two modes of rapid adenosine have been discovered. Spontaneous transients occur randomly in the brain, while mechanical stimulation also causes a rapid adenosine event. Pannexin1 channels are membrane channels that transport ions, including ATP, out of the cell where it is rapidly broken down into adenosine. Pannexin 1 channels (Panx1) have a flickering mode of rapid opening and are also mechanically stimulated. Here, we test the extent to which pannexin channels, specifically pannexin1 (Panx1) channels, are responsible for rapid adenosine events. Spontaneous adenosine release or mechanosensitive adenosine release were measured using fast-scan cyclic voltammetry in hippocampal (CA1) brain slices. In global Panx1KO mice, there is no significant difference in the frequency or concentration of spontaneous adenosine release, indicating Panx1 is not a release mechanism for spontaneous adenosine. Spontaneous adenosine frequency decreased slightly after administration of a large (100µM) dose of carbenoxolone, a nonspecific inhibitor of many pannexin and connexin channels, suggesting other hemichannels only play a small role at most. For mechanically stimulated adenosine release, the concentration of each adenosine event significantly decreased 30% in Panx1KO mice and the frequency of stimulations that evoked adenosine also decreased. The response was similar in WT mice with carbenoxolone. Thus, Panx1 is a release mechanism for mechanically stimulated adenosine release, but not the only mechanism. These results demonstrate that pannexin channels differentially regulate rapid adenosine release and could be targeted to differentially affect mechanically stimulated adenosine due to brain damage.

Using a simplified ATP algorithm to improve data reliability and improve cleanliness standards for surface and medical device hygiene.

An algorithm has been improved to mitigate variability in cleanliness measurements of various surfaces using rapid Adenosine Triphosphate (ATP) testing. A cleaning intervention step (CIS) verifies the cleanability of those surfaces. ATP testing was performed on surfaces which were pre-approved as "clean" and ready for re-use. Adjacent (duplicate) ATP sampling was undertaken on 421 environmental surfaces, medical devices and other implements. The CIS was conducted on 270 surfaces using an aseptic technique and disposable cleaning wipes. The two initial ATP results were plotted against each other with a 100 RLU threshold grading the results as clean (2x<100RLU), dirty (2x>100RLU) or equivocal (1x<100RLU and 1x>100RLU). Of the surfaces sampled, 68.5% were clean (288/421), 13.5% were dirty (57/421) and 18% were equivocal (76/421). The duplicate testing demonstrated a false negative rate of 10% (44/421) where the first swab was <100 RLU and the second swab >100 RLU. For the equivocal group, the gap between the two swabs was >100 RLU for 7.5% of surfaces (33/421). The CIS was conducted on 270 of the surfaces tested and showed that cleaning could be improved (P=<0.001) on 88.5% of surfaces (239/270). The simplified ATP testing algorithm provides real-time discrimination between surface cleanliness levels and improved certainty over surface hygiene. The duplicate swab sampling approach mitigates uncontrolled variability in the results and the CIS provides a nuanced understanding of the measurable cleanliness of any surface.

Dual-Channel Electrochemical Measurements Reveal Rapid Adenosine is Localized in Brain Slices.

Rapid adenosine signaling has been detected spontaneously or after mechanical stimulation in the brain, providing rapid neuromodulation in a local area. To measure rapid adenosine signaling, a single carbon-fiber microelectrode has traditionally been used, which limits spatial resolution and an understanding of regional coordination. In this study, we utilized dual-channel fast-scan cyclic voltammetry to measure the spontaneous or mechanically stimulated adenosine release at two electrodes placed at different spacings in hippocampal CA1 mouse brain slices. For mechanically stimulated adenosine release, adenosine can be detected up to 150 μm away from where it was stimulated, although the signal is smaller and delayed. While spontaneous adenosine transients were detected at both electrodes, only 10 percent of the events were detected concurrently, and that number was similar at 50 and 200 μm electrode spacings. Thus, most adenosine transients were not caused by the widespread coordination of release. There was no evidence of diffusion of spontaneous transients to a second electrode 50-200 μm away. This study shows that spontaneous adenosine events are very localized and thus provide only local neuromodulation. Injury, such as mechanical stimulation, allows adenosine to diffuse farther, but the neuroprotective effects are still regional. These results provide a better understanding of the spatial and temporal profiles of adenosine available to act at receptors, which is crucial for future studies that design neuroprotective treatments based on rapid adenosine signaling.

Clinical Efficacy of Novel Patient-Covering Negative-Pressure Box for Shielding Virus Transmission during Esophagogastroduodenoscopy: A Prospective Observational Study.

Esophagogastroduodenoscopy (EGD) has a high risk of virus transmission during the current coronavirus disease 2019 era, and preventive measures are under investigation. We investigated the effectiveness of a newly developed patient-covering negative-pressure box system (Endo barrier®) (EB) for EGD. Eighty consecutive unsedated patients who underwent screening EGD with EB use were prospectively enrolled. To examine the aerosol ratio before, during, and after EGD, 0.3- and 0.5-μm aerosols were measured every 60 s using an optical counter. Moreover, the degree of contamination of the examiners’ goggles and vinyl gowns was assessed before and after EGD using a rapid adenosine triphosphate (ATP) test for simulated droplets. Data were available in 73 patients and showed that 0.3- and 0.5-μm particles did not increase in 95.8% (70/73) and 94.5% (69/73) of patients during EGD under EB. There were no significant differences in the total 0.3- or 0.5-μm particle counts before versus after EGD. The difference in the ATP levels before and after EGD was −0.6 ± 16.6 relative light units (RLU) on goggles and 1.59 ± 19.9 RLU on gowns (both within the cutoff value). EB use during EGD may provide a certain preventive effect against aerosols and droplets, decreasing examiners’ exposure to viruses.

Facile electrochemical/colorimetric platform based on electrochromic tungsten oxide film for sensitive/visual adenosine triphosphate detection

In this paper, we present an electrochemical/colorimetric aptasensor based on tungsten oxide (WO3) film for rapid adenosine triphosphate (ATP) analysis. WO3 film was electrodeposited onto the surface of FTO substrate, and functioned in regulating conductivity and indicating color change of the aptasensor. For electrochemical impedance measurements, intercalation of Li+ ions in WO3 film could enhance its conductivity and thus decrease background signal. Compared with aptasensor under non-lithiated state, aptasensor under fully lithiated state exhibited a larger detection range, lower detection limit and superior selectivity. On the other hand, for colorimetric measurements, electrochromic behavior of the proposed aptasensor could be adjusted by the hinder effect induced by specific binding between aptamers and ATP. The proposed aptasensor provided both electrochemical and colorimetric detection modes, which combined the advantages of high sensitivity and simple readout. More importantly, this paper provides a simple strategy for enhancing the detection sensitivity of electrochemical sensors based on WO3 film, which holds a great significance in expanding the application of WO3 in electrochemical fields.

A1 and A2A Receptors Modulate Spontaneous Adenosine but Not Mechanically Stimulated Adenosine in the Caudate.

Adenosine is a neuromodulator, and rapid increases in adenosine in the brain occur spontaneously or after mechanical stimulation. However, the regulation of rapid adenosine by adenosine receptors is unclear, and understanding it would allow better manipulation of neuromodulation. The two main adenosine receptors in the brain are A1 receptors, which are inhibitory, and A2A receptors, which are excitatory. Here, we investigated the regulation of spontaneous adenosine and mechanically stimulated adenosine by adenosine receptors, using global A1 or A2A knockout mice. Results were compared in vivo and in brain slices' models. A1 KO mice have increased frequency of spontaneous adenosine events, but no change in the average concentration of an event, while A2A KO mice had no change in frequency but increased average event concentration. Thus, both A1 and A2A self-regulate spontaneous adenosine release; however, A1 acts on the frequency of events, while A2A receptors regulate concentration. The trends are similar both in vivo and slices, so brain slices are a good model system to study spontaneous adenosine release. For mechanically stimulated adenosine, there was no effect of A1 or A2A KO in vivo, but in brain slices, there was a significant increase in concentration evoked in A1KO mice. Mechanically stimulated release was largely unregulated by A1 and A2A receptors, likely because of a different release mechanism than spontaneous adenosine. Thus, A1 receptors affect the frequency of spontaneous adenosine transients, and A2A receptors affect the concentration. Therefore, future studies could probe drug treatments targeting A1 and A2A receptors to increase rapid adenosine neuromodulation.

A newly designed duodenoscope with detachable distal cap significantly reduces organic residue contamination after reprocessing.

A newly designed duodenoscope with detachable distal cap may reduce bacterial contamination by allowing better access to the elevator. We compared bacterial contamination and organic residue evaluated by rapid adenosine triphosphate (ATP) test and culture from duodenoscopes with detachable vs. fixed distal caps after high-level disinfection (HLD). During December 2018-April 2019, 108 used newly designed duodenoscopes were enrolled. In group A (n = 54), the distal cap of the duodenoscope was detached before manual cleaning. In group B (n = 54), the distal cap was not detached. After HLD, samples were collected from the elevator, submitted for culture, and evaluated using the ATP test, using the cutoff value of 40 relative light units (RLUs). After HLD, the proportion of potential bacterial contamination and organic residue in group A was significantly lower than in group B (37.0 % vs. 75.9 %; P < 0.001; relative risk 0.49, 95 % confidence interval 0.33-0.71), and also confirmed by median ATP values (45.2 vs. 141.0 RLU; P < 0.001). In group B, one sample culture was positive for nonpathogenic bacteria. Pathogenic bacteria were not found in any culture from either group. The detachable distal cap was more effective at eliminating bacterial contamination and reducing organic residue than a fixed cap. Nonpathogenic bacteria were detected in the fixed cap group after reprocessing. The ATP test with 40 RLU cutoff is a practical method to ensure the cleanliness of duodenoscope reprocessing without the need to wait for bacterial culture results.

CD73 or CD39 Deletion Reveals Different Mechanisms of Formation for Spontaneous and Mechanically Stimulated Adenosine and Sex Specific Compensations in ATP Degradation.

Adenosine is important for local neuromodulation, and rapid adenosine signaling can occur spontaneously or after mechanical stimulation, but little is known about how adenosine is formed in the extracellular space for those stimulations. Here, we studied mechanically stimulated and spontaneous adenosine to determine if rapid adenosine is formed by extracellular breakdown of adenosine triphosphate (ATP) using mice globally deficient in extracellular breakdown enzymes, either CD39 (nucleoside triphosphate diphosphohydrolase 1, NTPDase1) or CD73 (ecto-5'-nucleotidase). CD39 knockout (KO) mice have a lower frequency of spontaneous adenosine events than wild-type (WT, C57BL/6). Surprisingly, CD73KO mice demonstrate sex differences in spontaneous adenosine; males maintain similar event frequencies as WT, but females have significantly fewer events and lower concentrations. Examining the mRNA expression of other enzymes that metabolize ATP revealed tissue nonspecific alkaline phosphatase (TNAP) was upregulated in male CD73KO mice, but not secreted prostatic acid phosphatase (PAP) or transmembrane PAP. Thus, TNAP upregulation compensates for CD73 loss in males but not in females. These sex differences highlight that spontaneous adenosine is formed by metabolism of extracellular ATP by many enzymes. For mechanically stimulated adenosine, CD39KO or CD73KO did not change stimulation frequency, concentration, or t1/2. Thus, the mechanism of formation for mechanically stimulated adenosine is likely direct release of adenosine, different than spontaneous adenosine. Understanding these different mechanisms of rapid adenosine formation will help to develop pharmacological treatments that differentially target modes of rapid adenosine signaling, and all treatments should be studied in both sexes, given possible differences in extracellular ATP degradation.

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.

Comparison of spontaneous and mechanically-stimulated adenosine release in mice

Rapid adenosine signaling, on the time frame of seconds, has been discovered in the brain that can modulate neurotransmission or blood flow. Rapid adenosine release can occur spontaneously or be evoked after a mechanical stimulation, but these two modes of adenosine have not been compared. Here, we compared spontaneous and mechanically-stimulated adenosine release in the prefrontal cortex, striatum, and hippocampus of anesthetized mice. For spontaneous adenosine, the number of adenosine events in the prefrontal cortex (40 ± 4 per hour) was significantly lower than in the striatum (54 ± 3) or hippocampus (56 ± 3). Similarly, the concentration per transient was lower in the prefrontal cortex but highest in the striatum. For mechanically-stimulated adenosine, the peak concentration in the prefrontal cortex (8 ± 2 μM) and striatum (8 ± 1 μM) were significantly lower than in the hippocampus (16 ± 2 μM). Comparing the two modes, the hippocampus had high mechanically-stimulated concentration and high spontaneous frequency, while the prefrontal cortex had lower spontaneous frequency and mechanically-stimulated release. However, there is no pattern with the striatum and thus no direct correlations between spontaneous and mechanically-stimulated adenosine. Thus, there may be different pools of adenosine or mechanisms of formation for these two modes. Because of the high frequency of spontaneous events and high concentration of mechanically-stimulated release in the hippocampus, there may be some areas that have stronger adenosine signaling and thus stronger neuromodulatory control by adenosine.

Caffeine Modulates Spontaneous Adenosine and Oxygen Changes during Ischemia and Reperfusion.

Adenosine is an endogenous neuroprotectant that modulates vasodilation in the central nervous system. Oxygen changes occur when there is an increase in local cerebral blood flow and thus are a measure of vasodilation. Transient oxygen events following rapid adenosine events have been recently discovered, but the relationship between adenosine and blood flow change during ischemia/reperfusion (I/R) has not been characterized. Caffeine is a nonselective adenosine receptor antagonist that can modulate the effects of adenosine in the brain, but how it affects adenosine and oxygen levels during I/R is also unknown. In this study, extracellular changes in adenosine and oxygen were simultaneously monitored using fast-scan cyclic voltammetry during bilateral common carotid artery occlusion (BCCAO) and the effects of a specific A2A antagonist, SCH 442416, or general antagonist, caffeine, were studied. Measurements were made in the caudate-putamen for 1 h of normoxia, followed by 30 min of BCCAO and 30 min of reperfusion. The frequency and number of both adenosine and oxygen transient events significantly increased during I/R. The specific A2A antagonist, SCH 442416 (3 mg/kg, i.p.), eliminated the increase in adenosine and oxygen events caused by I/R. The general adenosine receptor antagonist, caffeine (100 mg/kg, i.p.), decreased the frequency of adenosine and oxygen transient events during I/R. These results demonstrate that, during BCCAO, there are more rapid release events of the neuromodulator adenosine and correlated local oxygen changes, and these rapid, local effects are dampened by caffeine and other A2A antagonists.

Early changes in transient adenosine during cerebral ischemia and reperfusion injury.

Adenosine is an important neuromodulator in the central nervous system, and tissue adenosine levels increase during ischemic events, attenuating excitotoxic neuronal injury. Recently, our lab developed an electrochemical fast-scan cyclic voltammetry (FSCV) method that identified rapid, spontaneous changes in adenosine concentrations that last only about 3 seconds. Here, we investigated the effects of cerebral ischemia and reperfusion on the concentration and frequency of transient adenosine release in the caudate-putamen. In anesthetized rats, data were collected for four hours: two hours of normoxia, 30 min of cerebral ischemia induced by bilateral common carotid artery occlusion, and 90 min of reperfusion. Transient adenosine release was increased during the cerebral ischemia period and remained elevated during reperfusion. The total number of adenosine transients increased by 52% during cerebral ischemia and reperfusion compared to normoxia. The concentration of adenosine per event did not increase but the cumulative adenosine concentration during cerebral ischemia and reperfusion increased by 53% because of the higher frequency of events. Further, we evaluated the role of A2A antagonist, SCH442416, a putative neuroprotective agent to affect adenosine transients. SCH442416 significantly decreased the transient frequency during cerebral ischemia-reperfusion by 27% and the cumulative concentration by 31%. Our results demonstrate that this mode of rapid adenosine release increases during early cerebral ischemia-reperfusion injury. Rapid adenosine release could provide fast, local neuromodulation and neuroprotection during cerebral ischemia.

A new sampling algorithm demonstrates that ultrasound equipment cleanliness can be improved

Australia has established guidelines on cleaning for reusable ultrasound probes and accompanying equipment. This is a preliminary study investigating cleanliness standards of patient-ready ultrasound equipment in 5 separate health care facilities within a major city. The cleanliness was assessed using rapid adenosine triphosphate (ATP) testing used with a sampling algorithm which mitigates variability normally associated with ATP testing. Each surface was initially sampled in duplicate for relative light units (RLUs) and checked for compliance with literature recommended levels of cleanliness (<100 RLUs). Triplicate sampling was undertaken where necessary. A cleaning intervention step (CIS) followed using a disposable detergent wipe, and the surface was retested for ATP. There were 253 surfaces tested from the 5 health care facilities with 26% (66/253) demonstrating either equivocal or apparent lack of cleanliness. The CIS was conducted on 148 surfaces and demonstrated that for >91% (135/148) of surfaces, the cleaning standards could be improved significantly (P > .001). For 6% (9/148) of devices and surfaces, the CIS needed to be repeated at least once to achieve the intended level of cleanliness (<25 RLUs). This study indicates that ATP testing is an effective, real-time, quality assurance tool for cleanliness monitoring of ultrasound probes and associated equipment.