287— The X-Ray Afterglows of Short Gamma-Ray Bursts

Light curves of the X-ray afterglow from 81 short gamma-ray bursts, from 2005 to 2018, were obtained from the Swift/XRT catalog. The light curves were fit with single power-law and double-power law models using the emcee library in Python and sorted into three groups: bursts well fit by a single power-law, bursts well fit by a double power-law, and bursts not well fit by either. Filters were applied to each category to identify bursts with a steep decay or a plateau. Once these unusual bursts were identified, their properties (duration, flux, fluence, and hardness) were compared to those of the more typical bursts. No significant trends were found to distinguish the unusual bursts from the rest

Investigating the Underlying Mechanisms Responsible for the Effectiveness of Behavioral Cough Therapy

Chronic cough (CC), a cough that lasts more than eight weeks, is the number one complaint of adults seeking non-emergent medical care. An estimated 20% of patients with CC do not respond to medical treatment, and are said to have refractory chronic cough (RCC). Several studies provide evidence that RCC is caused by hypersensitivity of sensory protein receptors in the airway epithelium known to regulate cough. The primary sensory receptor is the transient receptor potential vanilloid (TRPV). These receptors can be found in the epithelial layer of the bronchus, larynx and nose. These receptors are very plastic; in other words they are easily influenced by endogenous and exogenous stimulants. Behavioral cough therapy (BCT), which is provided by speech-language pathologists, has been shown to result in reduced cough sensitivity. However, the underlying mechanism that results in reduced cough sensitivity is unknown. We hypothesize the change is due to down-modulation of TRPV receptors through a neuroplastic mechanism. To test this hypothesis, we will use Western Blot analysis and quantitative polymerase chain reaction (qPCR) to estimate TRPV expression in biopsies of epithelial tissue obtained from the nose and larynx of patients pre and post-BCT. Examining the expression of TRPV before and after BCT will potentially explain the mechanism of the effect of BCT, which may increase its application in the clinic as well as open doors to other potential treatments for RCC

Investigating the Underlying Mechanisms Responsible for the Effectiveness of Behavioral Cough Suppression Therapy

Several studies provide evidence to support that refractory chronic cough (RCC) may be related to hypersensitivity of afferent receptors expressed on C-fibers in airway epithelium. The primary known sensory receptor involved in regulation of cough sensitivity is the transient receptor potential vanilloid 1 (TRPV1). TRPV1 receptors are found in the epithelial layer of the bronchus, larynx, and nose, and are very plastic. In other words, they are easily influenced by endogenous and exogenous stimulants. Behavioral cough suppression therapy (BCST) has been shown to improve RCC symptoms in approximately 80% of patients. Several studies have shown a change in cough sensitivity following successful BCST, as evidenced by a change in capsaicin cough-challenge testing. However, the underlying mechanism that results in reduced cough sensitivity is unknown. Given that capsaicin is a known agonist of the TRPV1 receptor, and the high degree of plasticity of TRPV1 receptors, we hypothesize that the improvement seen after BCST is due to down-modulation of TRPV1 receptors through a neuroplastic mechanism. To test this hypothesis, we will use Western Blot analysis and quantitative polymerase chain reaction (qPCR) to quantify TRPV1 expression in biopsies of epithelial tissue obtained from the nose and larynx (epiglottis) of patients with RCC pre and post-BCST. An improved understanding of the mechanism responsible for the treatment effect of BCST may help to increase its application in the clinical setting as well as open doors to other potential treatments for RCC. The University of Montana Institutional Review Board has approved this study. Participant recruitment is currently underway. We anticipate having data on up to four participants by Fall Voice 2019. Keywords: refractory chronic cough; behavioral cough suppression therapy; transient receptor potential vanilloid 1 (TRPV1

Patient-Reported Variables Associated with the Success of Behavioral Intervention for Patients with Chronic Cough

Chronic cough is a common condition that persists for more than eight weeks and accounts for millions of visits to physicians each year. Approximately 10-20% of patients with chronic cough do not respond to medical treatment. These patients, said to have refractory chronic cough (RCC), often respond to behavioral cough suppression therapy (BCST), provided by a speech-language pathologist (SLP). The purpose of this study was to determine if common factors exist that distinguish patients with RCC, who benefit from BCST from those who don’t benefit from BCST. The long-term goal of this research is to create a screening tool that physicians can use to identify candidates for BCST. Forty three adults referred for BCST completed an enrollment survey at the beginning of therapy. The survey consisted of 52-items pertaining to cough onset factors, cough symptoms, and personality traits. Participants also completed the Leicester Cough Questionnaire (LCQ), a validated measurement for assessing cough-related quality of life. A follow- up survey with similar questions regarding their cough status and changes in treatment was administered 3-4 weeks after beginning BCST. Thirty participants were satisfied following BCST (BCST-S); 13 were not satisfied (BCST-NS). There were significant differences between the two groups on: cough productivity, tight-throat feeling, symptoms of reflux, and stress as a cough trigger. Additionally patients in the BCST-S group described themselves as significantly more anxious and stressed than the patients in the BCST-NS group. The LCQ confirmed a significant improvement in the BCST-S group, but not the BCST-NS group. The study indicates potential to create a valid screening tool that would assist with the identification of candidates for BCST. Such a tool could save considerable time and money for patients with RCC

Impact of preanalytical factors on the concentration of specific metabolic hormones and effect of age and fasting on specific metabolic hormones in rats

Die biochemische Analyse von Hormonen wird durch drei Variablen beeinflusst – die präanalytischen, analytischen und postanalytischen Variabilität. Bezüglich der analytischen und biologischen Variabilität wurden für Nager schon zahlreiche Arbeiten veröffentlicht, jedoch ist die Präanalytik in der Untersuchung von Blutproben von Ratten, in Bezug auf die Hormonanalytik, bislang kaum beachtet worden. Deswegen wurden in dieser Arbeit einige Aspekte der Präanalytik bei der Hormonmessung in Ratten genauer untersucht; auch um Anhaltspunkte zu geben, in wie weit vorbehandelte Blutproben von Ratten zur Messung unterschiedlicher Hormone genutzt werden können und wie sich die Verwendung unterschiedlicher Probenmaterialien, als auch Einfrier-Auftau-Zyklen, auf die Messergebnisse verschiedener Stoffwechselhormone auswirken können. In Bezug auf die biologische Variabilität wurde der Einfluss von Alter und Fasten auf ausgewählte Stoffwechselhormone bei der Ratte genauer untersucht. Ziel dieser Arbeit war es, in Analyseprozessen insbesondere auf den Faktor „Präanalytik“ einzugehen und aufzuzeigen, in welchem Ausmaß Ergebnisse in der Messung von Hormonen mittels Immunassays beeinflusst werden können. Im Vergleich zu reinem Serum waren die gemessenen Konzentrationen von IGF-I (+9,2%, p<0,001), IGF-II (+24,0%, p<0,001), IGFBP-3 (+24,0%, p<0,001) und Leptin (+54,9%, p<0,0001) in EDTA-Plasma signifikant niedriger, GH (-137,8%, p<0,001) und Gesamt-Ghrelin (-10,8%, p<0,05) zeigten erhöhte Werte, für IGFBP- 2, Gesamt-GIP, aktives GLP-1, Insulin und Gesamt-Ghrelin zeichneten sich keine Differenzen ab. Nach zehn Einfrier-Auftau-Zyklen von Serum blieben die Konzentrationen von GH, IGF-I, IGFBP-2 und aktivem GLP-1 unverändert, bei Gesamt-GIP (+49,8%, p<0,001), Insulin (+32,9%, p<0,001) und Gesamt-Ghrelin (+24,6%, p<0,001), konnte eine geringere Menge detektiert werden und bei IGF-II (-25,9%, p<0,01), IGFBP-3 (-19,3%, p<0,001) und Leptin (-41,3%, p<0,001) wurden höhere Werte gemessen. EDTA-Plasmaproben, die ab dem Moment der Abnahme von der Ratte bis zum gefrorenen Aufbewahren ununterbrochen gekühlt wurden, zeigten keine signifikanten Unterschiede zu solchen Proben, die bei Raumtemperatur bis zum Einfrierprozess aufbewahrt wurden. Die Zugabe von Proteaseinhibitoren, im Speziellen von DPP-4-Inhibitor, ist für die Bestimmung von Inkretinhormonen angezeigt. In dieser Studie wurde keine erhöhte Variabilität durch den Zusatz der Proteaseinhibitoren auf die Messung von Gesamt-GIP und aktivem GLP-1 gefunden. Für die Messung von acyliertem Ghrelin ist eine Ansäuerung des Probenmaterials mit HCl empfehlenswert. IGF-II, IGFBP-2, IGFBP-3, Gesamt-GIP und Insulin konnten mit dieser Probenvorbehandlung bedenkenlos gemessen werden. Für Leptin (-73,91%, p<0,05) wurden erhöhte Konzentrationen detektiert, IGF-I (+11,9%, p<0,001) und Gesamt-Ghrelin (+27,1%, p<0,05) zeigten hingegen verringerte Werte. In Bezug auf die biologische Variabilität konnte ebenfalls ein signifikanter Einfluss des Alters und Fastens auf Ratten detektiert werden. Jungtieren hatten einen deutlicher niedrigeren Insulinspiegel (+0,22%, p<0,05) im Blut, bei einer Fastendauer von 16 Stunden, im Vergleich zu wesentlich älteren 1-Jahr alten Ratten. Im Vergleich der Insulinwerte zwischen 6 und 16 Stunden des Fastens konnten signifikant niedrigere Werte nach 16 Stunden festgestellt werden (+30,7, p<0,05). Bei jungen Ratten reduzierte sich die Gesamt-GIP-Konzentration (+105,9%, p<0,01) während des Fastens stärker als bei alten Tieren, aber bei diesem Hormon ist, bei Langzeitfasten aller Tiere, eine signifikante Reduktion der Gesamt- GIP-Konzentration (66,8%, p<0,05) im Vergleich von 6 zu 16 Stunden zu verzeichnen gewesen. Bei IGF-I und Gesamt-Ghrelin veränderten sich die Konzentrationen der zirkulierenden Hormone im Blut trotz einer Fastendauer von 16 Stunden nicht signifikant. Es konnten in dieser Arbeit einige Faktoren der Präanalytik, im Speziellen in der Messung von Hormonen mittels Immunassays bei Ratten, untersucht werden, jedoch sind auf diesem Themengebiet noch zahlreiche weitere Fragestellungen, wie zum Beispiel der Einfluss von Einfrier-Auftau-Zyklen auf EDTA-Plasma oder der Einfluss von Matrixeffekten auf Western Liganden Blot, noch nicht näher untersucht, weswegen es sinnvoll erscheint diesbezüglich weitere Studien anzuschließen.Biochemical analysis of hormones is influenced by three factors – preanalytical, analytical and postanalytical variability. For analytical und biological variability many studies have been published, but the preanalytical measurement of hormones is less examined for blood samples in rats. Therefore we investigated in some issues of preanalytical hormone measurement, to give evidence, if pretreated blood samples also can be used for measuring different hormones and how the usage of different blood sample materials (serum or EDTA-plasma) can affect the measured concentrations of different metabolic hormones. Furthermore the influence of freeze and thawing cycles on the concentrations of different metabolic hormones has been studied. We also investigated in two compounds of the biological variability: the influence of age and fasting on specific metabolic hormones in rats. With this study we aimed to point out the factor “preanalytic“ in the analytical processes and we were able to show how much the results of hormone measurement, detected by immunassays, can be influenced by these components. In relation to plain serum the concentrations of IGF-I (+9.2%, p<0.001), IGF-II (+24.0%, p<0.001), IGFBP-3 (+24.0%, p<0.001) and leptin (+54.9%, p<0.0001) had significantly lower levels in EDTA-plasma whereas GH (-137.8%, p<0.001) and total Ghrelin (-10.8%, p<0.05) showed significant higher levels and for IGFBP- 2, total GIP, active GLP-1, insulin and total Ghrelin there has been no significant difference. After ten freezing and thawing cycles of serum the concentrations of GH, IGF-I, IGFBP-2 and active GLP-1 have not changed, however lower concentrations have been detected for total GIP (+49.8%, p<0.001), insulin (+32.9%, p<0.001) and total Ghrelin (+24.6%, p<0.001). For IGF-II (-25.9%, p<0.01), IGFBP-3 (-19.3%, p<0.001) and leptin (-41.3%, p<0.001) higher concentrations have been measured after ten freezing and thawing cycles. Comparing blood samples, kept at room temperature during treatment until getting frozen for long-time storage, to those blood samples, which have been stored on ice until long-time storage there have no significant differences been detected. The addition of protease inhibitors, especially the addtiton of DPP-4-inhibitor, is advised for the measuring of incretins. In this study no higher variability in case of adding protease inhibitors to blood samples has been detected in the measurement of total GIP and active GLP-1. The addition of HCl to blood samples is recommended for measuring active Ghrelin, but IGF-II, IGFBP-2, IGFBP-3, total GIP and insulin can also be measured from this pretreated blood samples without deviation in their hormone concentrations. After addition of HCl for leptin (- 73.91%, p<0.05) higher concentrations have been detected and for IGF-I (+11.9%, p<0.001) and total Ghrelin (+27.1%, p<0.05) lower concentrations have been measured. As a component of the biological variability, there has also been a significant influence of age and fasting in rats. Young rats (14 weeks old) had lower levels of insulin (0.22%, p<0.05) after fasting for 16h in relation to one year old rats. When comparing insulin concentrations after 6 and 16h of fasting there have been significant lower levels after 16h (+30.7%, p<0.05). While fasting, the concentration of total GIP (+105.9%, p<0.01) was more reduced in young rats than in old ones. Furthermore there was a significant reduction of total GIP after 6 to 16h of fasting (66.8%, p<0.05). For IGF-I and total Ghrelin there was no difference in fasting the rats for 6 or 16h. In this study we investigated some aspects of preanalytic, especially in the measurement of circulating metabolic hormones by immunoassay in rats. However there are still much more topics to be researched, like the influence of freezing and thawing cycles on hormone concentrations in EDTA-plasma or the impact of matrix effects on Western ligand blot

An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria

Owing to their small size and enhanced stability, nanobodies derived from camelids have previously been used for the construction of intracellular "nanotraps," which enable redirection and manipulation of green fluorescent protein (GFP)-tagged targets within living plant and animal cells. By taking advantage of intracellular compartmentalization in the magnetic bacterium Magnetospirillum gryphiswaldense, we demonstrate that proteins and even entire organelles can be retargeted also within prokaryotic cells by versatile nanotrap technology. Expression of multivalent GFP-binding nanobodies on magnetosomes ectopically recruited the chemotaxis protein CheW(1)-GFP from polar chemoreceptor clusters to the midcell, resulting in a gradual knockdown of aerotaxis. Conversely, entire magnetosome chains could be redirected from the midcell and tethered to one of the cell poles. Similar approaches could potentially be used for building synthetic cellular structures and targeted protein knockdowns in other bacteria. IMPORTANCE Intrabodies are commonly used in eukaryotic systems for intracellular analysis and manipulation of proteins within distinct subcellular compartments. In particular, so-called nanobodies have great potential for synthetic biology approaches because they can be expressed easily in heterologous hosts and actively interact with intracellular targets, for instance, by the construction of intracellular "nanotraps" in living animal and plant cells. Although prokaryotic cells also exhibit a considerable degree of intracellular organization, there are few tools available equivalent to the well-established methods used in eukaryotes. Here, we demonstrate the ectopic retargeting and depletion of polar membrane proteins and entire organelles to distinct compartments in a magnetotactic bacterium, resulting in a gradual knockdown of magneto-aerotaxis. This intracellular nanotrap approach has the potential to be applied in other bacteria for building synthetic cellular structures, manipulating protein function, and creating gradual targeted knockdowns. Our findings provide a proof of principle for the universal use of fluorescently tagged proteins as targets for nanotraps to fulfill these tasks

A minisandwich experiment with blended ca-bentonite and pearson water—Hydration, swelling, solute transport and cation exchange

Shaft seals are geotechnical barriers in nuclear waste deposits and underground mines. The Sandwich sealing system consists of alternating sealing segments (DS) of bentonite and equipotential segments (ES). MiniSandwich experiments were performed with blended Ca-bentonite (90 mm diameter and 125 mm height) to study hydration, swelling, solute transport and cation exchange during hydration with A3 Pearson water, which resembles pore water of Opalinus Clay Formation at sandy facies. Two experiments were run in parallel with DS installed either in one-layer hydrate state (1W) or in air-dry two-layer hydrate (2W) state. Breakthrough at 0.3 MPa injection pressure occurred after 20 days and the fluid inlet was closed after 543 days, where 4289 mL and 2984 mL, respectively, passed both cells. Final hydraulic permeability was 2.0–2.7 × 10$^{-17}$ m$^{2}$. Cells were kept for another 142 days before dismantling. Swelling of DS resulted in slight compaction of ES. No changes in the mineralogy of the DS and ES material despite precipitated halite and sulfates occurred. Overall cation exchange capacity of the DS does not change, maintaining an overall value of 72 ± 2 cmol(+)/kg. Exchangeable Na$^{+}$ strongly increased while exchangeable Ca$^{2+}$ decreased. Exchangeable Mg$^{2+}$ and K$^{+}$ remained nearly constant. Sodium concentration in the outflow indicated two different exchange processes while the concentration of calcium and magnesium decreased potentially. Concentration of sulfate increased in the outflow, until it reached a constant value and chloride concentration decreased to a minimum before it slightly increased to a constant value. The available data set will be used to adapt numerical models for a mechanism-based description of the observed physical and geochemical processes