We demonstrate the density and shape of platinum nanoparticles (PtNP) on carbon-fiber microelectrodes with fast-scan cyclic voltammetry (FSCV) directly impacts detection of adenosine. Previously, we ...showed that metal nanoparticle-modified carbon significantly improves adenine-based purine detection; however, how the size and shape of the particles impact electrochemical detection was not investigated. Electrochemical investigations of how the surface topology and morphology impacts detection is necessary for designing ultrasensitive electrodes and for expanding fundamental knowledge of electrode-analyte interactions. To change the density and shape of the PtNP's on the surface, we varied the concentration of K
PtCl
and electrodeposition time. We show that increasing the concentration of K
PtCl
increases the density of PtNP's while increasing the electrodeposition time impacts both the density and size. These changes manipulate the adsorption behavior which impacts sensitivity. Based on these results, an optimal electrodeposition procedure was determined to be 1.0 mg/mL of K
PtCl
deposited for 45 s and this results in an average increase in adenosine detection by 3.5 ±0.3-fold. Interestingly, increasing the size and density of PtNPs negatively impacts dopamine detection. Overall, this work provides fundamental insights into the differences between adenosine and dopamine interaction at electrode surfaces.
We introduce a portable biochemical analysis platform for rapid field deployment of nucleic acid-based diagnostics using consumer-class quadcopter drones. This approach exploits the ability to ...isothermally perform the polymerase chain reaction (PCR) with a single heater, enabling the system to be operated using standard 5 V USB sources that power mobile devices (via battery, solar, or hand crank action). Time-resolved fluorescence detection and quantification is achieved using a smartphone camera and integrated image analysis app. Standard sample preparation is enabled by leveraging the drone’s motors as centrifuges via 3D printed snap-on attachments. These advancements make it possible to build a complete DNA/RNA analysis system at a cost of ∼$50 ($US). Our instrument is rugged and versatile, enabling pinpoint deployment of sophisticated diagnostics to distributed field sites. This capability is demonstrated by successful in-flight replication of Staphylococcus aureus and λ-phage DNA targets in under 20 min. The ability to perform rapid in-flight assays with smartphone connectivity eliminates delays between sample collection and analysis so that test results can be delivered in minutes, suggesting new possibilities for drone-based systems to function in broader and more sophisticated roles beyond cargo transport and imaging.
Chronic myelomonocytic leukemia (CMML) is a myeloid neoplasm characterized by dysplasia, abnormal production and accumulation of monocytic cells and an elevated risk of transforming into acute ...leukemia. Over the past two decades, our knowledge about the pathogenesis and molecular mechanisms in CMML has increased substantially. In parallel, better diagnostic criteria and therapeutic strategies have been developed. However, many questions remain regarding prognostication and optimal therapy. In addition, there is a need to define potential pre-phases of CMML and special CMML variants, and to separate these entities from each other and from conditions mimicking CMML. To address these unmet needs, an international consensus group met in a Working Conference in August 2018 and discussed open questions and issues around CMML, its variants, and pre-CMML conditions. The outcomes of this meeting are summarized herein and include diag nostic criteria and a proposed classification of pre-CMML conditions as well as refined minimal diagnostic criteria for classical CMML and special CMML variants, including oligomonocytic CMML and CMML associated with systemic mastocytosis. Moreover, we propose diagnostic standards and tools to distinguish between 'normal', pre-CMML and CMML entities. These criteria and standards should facilitate diagnostic and prognostic evaluations in daily practice and clinical studies in applied hematology.
In this paper, we describe the investigations and actions taken to reduce risk and prevent casualties from a catastrophic 210,000 m
3
rockslope failure, which occurred near the village of Preonzo in ...the Swiss Alps on May 15, 2012. We describe the geological predisposition and displacement history before and during the accelerated creep stage as well as the development and operation of an efficient early warning system. The failure of May 15, 2012, occurred from a large and retrogressive instability in gneisses and amphibolites with a total volume of about 350,000 m
3
, which formed an alpine meadow 1250 m above the valley floor. About 140,000 m
3
of unstable rock mass remained in place and might collapse partially or completely in the future. The instability showed clearly visible signs of movements along a tension crack since 1989 and accelerated creep with significant hydromechanical forcing since about 2006. Because the active rockslide at Preonzo threatened a large industrial facility and important transport routes located directly at the toe of the slope, an early warning system was installed in 2010. The thresholds for prealarm, general public alarm, and evacuation were derived from crack meter and total station monitoring data covering a period of about 10 years, supplemented with information from past failure events with similar predisposition. These thresholds were successfully applied to evacuate the industrial facility and to close important roads a few days before the catastrophic slope failure of May 15, 2012. The rock slope failure occurred in two events, exposing a compound rupture plane dipping 42° and generating deposits in the midslope portion with a travel angle of 39°. Three hours after the second rockslide, the fresh deposits became reactivated in a devastating debris avalanche that reached the foot of the slope but did not destroy any infrastructure. The final run-out distance of this combined rock collapse–debris avalanche corresponded to the predictions made in the year 2004.
Leukemic stem cells (LSCs) are an emerging target of curative anti-leukemia therapy. In acute lymphoblastic leukemia (ALL), LSCs frequently express CD34 and often lack CD38. However, little is known ...about markers and targets expressed in ALL LSCs. We have examined marker- and target expression profiles in CD34+/CD38− LSCs in patients with Ph+ ALL (n = 22) and Ph− ALL (n = 27) by multi-color flow cytometry and qPCR. ALL LSCs expressed CD19 (B4), CD44 (Pgp-1), CD123 (IL-3RA), and CD184 (CXCR4) in all patients tested. Moreover, in various subgroups of patients, LSCs also displayed CD20 (MS4A1) (10/41 = 24%), CD22 (12/20 = 60%), CD33 (Siglec-3) (20/48 = 42%), CD52 (CAMPATH-1) (17/40 = 43%), IL-1RAP (13/29 = 45%), and/or CD135 (FLT3) (4/20 = 20%). CD25 (IL-2RA) and CD26 (DPPIV) were expressed on LSCs in Ph+ ALL exhibiting BCR/ABL1p210, whereas in Ph+ ALL with BCR/ABL1p190, LSCs variably expressed CD25 but did not express CD26. In Ph− ALL, CD34+/CD38− LSCs expressed IL-1RAP in 6/18 patients (33%), but did not express CD25 or CD26. Normal stem cells stained negative for CD25, CD26 and IL-1RAP, and expressed only low amounts of CD52. In xenotransplantation experiments, CD34+/CD38− and CD34+/CD38+ cells engrafted NSG mice after 12–20 weeks, and targeting with antibodies against CD33 and CD52 resulted in reduced engraftment. Together, LSCs in Ph+ and Ph− ALL display unique marker- and target expression profiles. In Ph+ ALL with BCR/ABL1p210, the LSC-phenotype closely resembles the marker-profile of CD34+/CD38− LSCs in chronic myeloid leukemia, confirming the close biologic relationship of these neoplasms. Targeting of LSCs with specific antibodies or related immunotherapies may facilitate LSC eradication in ALL.
Abstract
Norepinephrine (NE) is a ubiquitous neurotransmitter that facilitates cellular communication in the sympathetic nervous system. Recent advancements have shown sympathetic neurons innervate ...secondary immune organs, such as the spleen, to control immune physiology. Fast-scan cyclic voltammetry (FSCV) is an electrochemical technique that varies an applied potential at a very fast scan rate with micro-electrodes to detect neurotransmitters with high temporal and spatial resolution. FSCV can be used to detect the sub-second release of NE in an ex vivomurine slice model to study the mechanisms related to neuro-immune communication. Our lab is the first to characterize and quantify this spontaneous transient signaling of catecholamines in the white pulp region of the murine spleen with FSCV. The average concentration of catecholamines released is 581 ± 20 nM, with an average inter-event time of 23.4 ± 0.7 s and a duration of 6.6 ± 0.1 s. Norepinephrine transporter (NET) is known to transport NE back into cells after endogenous release. We blocked NET with the inhibitor nomifensine to provide further evidence of NE release and study the mechanisms involved in NE uptake. To study the mechanisms of NE release in the spleen, we blocked α2-adrenoceptor, a known autoreceptor only found on neurons. Yohimbine is an α2-adrenoceptor antagonist, and when applied to slices, it increased catecholamine concentration and duration, demonstrating that we are detecting neuronal release of NE. Furthermore, we used ELISAs to quantify the extent to which the α2-adrenoceptor affects cytokine production, providing further evidence neuronal-regulated immune signaling in the white pulp region of the spleen.
Supported by grants from NIH (R01AI151552, R01NS121426)
Prediction of a boar's fertility level has great economic importance for sow herds. After standard sperm morphology and motility metrics are met, approximately 25% of boars have less than 80% ...conception rates. Due in part to the many factors involved in the fertilization process, a multifactorial model incorporating multiple relevant sperm physiology factors will likely lead to increased understanding of boar fertility. Here we review the current literature on boar sperm capacitation as a predictor of boar fertility. While limited, several studies have provided correlations between the percentage of sperm in an ejaculate that are capable of undergoing sperm capacitation in a chemically defined media and artificial insemination field fertility as well as proteome and other methods. Work summarized here underscores the need for further understanding of boar fertility.
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