Emerging key roles for P2X receptors in the kidney

P2X ionotropic non-selective cation channels are expressed throughout the kidney and are activated in a paracrine or autocrine manner following the binding of extracellular ATP and related extracellular nucleotides. Whilst there is a wealth of literature describing a regulatory role of P2 receptors (P2R) in the kidney, there are significantly less data on the regulatory role of P2X receptors (P2XR) compared with that described for metabotropic P2Y. Much of the historical literature describing a role for P2XR in the kidney has focused heavily on the role of P2X1R in the autoregulation of renal blood flow. More recently, however, there has been a plethora of manuscripts providing compelling evidence for additional roles for P2XR in both kidney health and disease. This review summarizes the current evidence for the involvement of P2XR in the regulation of renal tubular and vascular function, and highlights the novel data describing their putative roles in regulating physiological and pathophysiological processes in the kidney

Better Pumps: Reliable Handpump Infrastructure

Approximately 90 million people in Africa lack access to safe drinking water, despite having water infrastructure installed in their community. The India Mark II and the Afridev handpumps are among the most widely used handpumps in the world. Sadly, studies show that approximately 30% of these handpumps are non-operational due to failures of the bearings, seals, head flange, and other common components. The Better Pumps team of the Collaboratory provides engineering support for partners who are working to improve handpump sustainability. We partnered with Tony Beers and AlignedWorks to validate a bearing test methodology for the India Mark II handpump. By modifying the loading conditions in our handpump test machine, we were able to replicate failures observed by AlignedWorks in a field trial of their bearing design. We partnered with Matt Schwiebert and Living Water International to test new seal designs for the India Mark II and Afridev handpumps and to measure head flange deflections in the India Mark II handpump. Seal performance data collected by the team was used to validate a new design in advance of field trials by Living Water International. Head flange deflection data was collected for partner benchmarking of their computational analysis. Test methodologies and results are reported.https://mosaic.messiah.edu/engr2021/1000/thumbnail.jp

Pediatric resident and faculty attitudes toward self-assessment and self-directed learning: a cross-sectional study

<p>Abstract</p> <p>Background</p> <p>The development of self-assessment and self-directed learning skills is essential to lifelong learning and becoming an effective physician. Pediatric residents in the United States are now required to use Individualized Learning Plans (ILPs) to document self-assessment and self-directed learning. A better understanding of resident and faculty attitudes and skills about self-assessment and self-directed learning will allow more successful integration of lifelong learning into residency education. The objective of this study was to compare faculty and resident attitudes, knowledge and skills about self-assessment, self-directed learning and ILPs.</p> <p>Methods</p> <p>Survey of pediatric residents and faculty at a single institution. Respondents rated their attitudes, knowledge, and self-perceived skills surrounding self-assessment, self-directed learning and ILPs.</p> <p>Results</p> <p>Overall survey response rate was 81% (79/97); 100% (36/36) residents and 70% (43/61) faculty. Residents and faculty agreed that lifelong learning is a necessary part of being a physician. Both groups were comfortable with assessing their own strengths and weaknesses and developing specific goals to improve their own performance. However, residents were less likely than faculty to continuously assess their own performance (44% vs. 81%; p < 0.001) or continuously direct their own learning (53% vs. 86%; p < 0.001). Residents were more likely than faculty to believe that residents should be primarily responsible for directing their own learning (64% vs. 19%; p < 0.0001), but at the same time, more residents believed that assigned clinical (31% vs. 0%; p < 0.0001) or curricular (31% vs. 0%; p < 0.0001) experiences were sufficient to make them competent physicians. Interns were less likely than senior residents to have a good understanding of how to assess their own skills (8% vs. 58%; p = 0.004) or what it means to be a self-directed learner (50% vs. 83%; p = 0.04).</p> <p>Qualitative comments indicated that while ILPs have the potential to help learners develop individualized, goal-directed learning plans based on strengths and weaknesses, successful implementation will require dedicated time and resident and faculty development.</p> <p>Conclusion</p> <p>These findings suggest that training and experience are necessary for physicians to understand the role of self-directed learning in education. Deliberate practice, for example by requiring residents to use ILPs, may facilitate self-directed, lifelong learning.</p

Deletion of airway cilia results in noninflammatory bronchiectasis and hyperreactive airways

The mechanisms for the development of bronchiectasis and airway hyperreactivity have not been fully elucidated. Although genetic, acquired diseases and environmental influences may play a role, it is also possible that motile cilia can influence this disease process. We hypothesized that deletion of a key intraflagellar transport molecule, IFT88, in mature mice causes loss of cilia, resulting in airway remodeling. Airway cilia were deleted by knockout of IFT88, and airway remodeling and pulmonary function were evaluated. In IFT88− mice there was a substantial loss of airway cilia on respiratory epithelium. Three months after the deletion of cilia, there was clear evidence for bronchial remodeling that was not associated with inflammation or apparent defects in mucus clearance. There was evidence for airway epithelial cell hypertrophy and hyperplasia. IFT88− mice exhibited increased airway reactivity to a methacholine challenge and decreased ciliary beat frequency in the few remaining cells that possessed cilia. With deletion of respiratory cilia there was a marked increase in the number of club cells as seen by scanning electron microscopy. We suggest that airway remodeling may be exacerbated by the presence of club cells, since these cells are involved in airway repair. Club cells may be prevented from differentiating into respiratory epithelial cells because of a lack of IFT88 protein that is necessary to form a single nonmotile cilium. This monocilium is a prerequisite for these progenitor cells to transition into respiratory epithelial cells. In conclusion, motile cilia may play an important role in controlling airway structure and function

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

Nucleotides and nucleosides are not only involved in cellular metabolism but also act extracellularly via P1 and P2 receptors, to elicit a wide variety of physiological and pathophysiological responses through paracrine and autocrine signalling pathways. For the first time, we have used an ion-pair reversed-phase high-performance liquid chromatography ultraviolet (UV)-coupled method to rapidly and simultaneously quantify 12 different nucleotides and nucleosides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine, uridine triphosphate, uridine diphosphate, uridine monophosphate, uridine, guanosine triphosphate, guanosine diphosphate, guanosine monophosphate, guanosine): (1) released from a mouse renal cell line (M1 cortical collecting duct) and (2) in human biological samples (i.e., urine). To facilitate analysis of urine samples, a solid-phase extraction step was incorporated (overall recovery rate ? 98 %). All samples were analyzed following injection (100 ?l) into a Synergi Polar-RP 80 Å (250 × 4.6 mm) reversed-phase column with a particle size of 10 ?m, protected with a guard column. A gradient elution profile was run with a mobile phase (phosphate buffer plus ion-pairing agent tetrabutylammonium hydrogen sulfate; pH 6) in 2-30 % acetonitrile (v/v) for 35 min (including equilibration time) at 1 ml min(-1) flow rate. Eluted compounds were detected by UV absorbance at 254 nm and quantified using standard curves for nucleotide and nucleoside mixtures of known concentration. Following validation (specificity, linearity, limits of detection and quantitation, system precision, accuracy, and intermediate precision parameters), this protocol was successfully and reproducibly used to quantify picomolar to nanomolar concentrations of nucleosides and nucleotides in isotonic and hypotonic cell buffers that transiently bathed M1 cells, and urine samples from normal subjects and overactive bladder patients

Purinergic inhibition of Na+,K+,Cl− cotransport in C11-MDCK cells: Role of stress-activated protein kinases

Previously, we observed that sustained activation of P2Y1 leads to inhibition of Na+,K+,Cl− cotransport (NKCC) in C11 cells resembling intercalated cells from collecting ducts of the Madin-Darby canine kidney. This study examined the role of stress-activated protein kinases (SAPK) in NKCC inhibition triggered by purinergic receptors. Treatment of C11 cells with ATP led to sustained phosphorylation of SAPK such as JNK and p38. Activation of these kinases also occurred in anisomycin-treated cells. Surprisingly, we observed that compounds SP600125 and SB202190, known as potent inhibitors of JNK and p38 in cell-free systems, activated rather than inhibited phosphorylation of the kinases in C11 cells. Importantly, similarly to ATP, all the above-listed activators of JNK and p38 phosphorylation inhibited NKCC. Thus, our results suggest that activation of JNK and/or p38 contributes to NKCC suppression detected in intercalated-like cells from distal tubules after their exposure to P2Y1 agonists

A case of serendipity*

An account is given of how a sensitive bioassay system for measurement of the neurotransmitter acetylcholine serendipitously led to the identification of adenosine triphosphate (ATP) released in vitro from active skeletal muscle. Subsequent application of the identification procedures to exercising human muscle in vivo, cardiac muscle cells in vitro, and human erythrocytes exposed to hypoxia gave rise to the general concept of ATP as a molecule that could influence cell function from the extracellular direction. Mechanisms of ATP release from cells in terms of “trigger” events such as mechanical distortion of the membrane, depolarization of the membrane, and exposure to hypoxia are discussed. Potential therapeutic uses of extracellular ATP in cancer therapy, radiation therapy, and a possible influence upon aging are discussed. Possible roles (distant and local) of extracellular ATP released from muscle during whole body exercise are discussed