Here we present two robotic sample changers integrated into the experimental stations for the macromolecular crystallography (MX) beamlines AMX and FMX, and the biological small‐angle scattering ...(bioSAXS) beamline LiX. They enable fully automated unattended data collection and remote access to the beamlines. The system designs incorporate high‐throughput, versatility, high‐capacity, resource sharing and robustness. All systems are centered around a six‐axis industrial robotic arm coupled with a force torque sensor and in‐house end effectors (grippers). They have the same software architecture and the facility standard EPICS‐based BEAST alarm system. The MX system is compatible with SPINE bases and Unipucks. It comprises a liquid nitrogen dewar holding 384 samples (24 Unipucks) and a stay‐cold gripper, and utilizes machine vision software to track the sample during operations and to calculate the final mount position on the goniometer. The bioSAXS system has an in‐house engineered sample storage unit that can hold up to 360 samples (20 sample holders) which keeps samples at a user‐set temperature (277 K to 300 K). The MX systems were deployed in early 2017 and the bioSAXS system in early 2019.
Robotic sample changers for macromolecular crystallography and biological small‐angle X‐ray scattering are presented.
This study examines the hypothesis that cholecystitis down-regulates Guinea pig gallbladder (GPGB) smooth muscle cholecystokinin (CCK)-stimulated prostaglandin (PG) release. Guinea pig gallbladder ...from Control and 48 h bile duct ligated (BDL) animals were placed in cell culture and grown to confluence. The cultures underwent Western Blot analysis for smooth muscle cell content of COX-1, COX-2, Prostacyclin Synthase (PS), or were incubated with CCK at 10(-8)M or 10(-6)M with and without indomethacin for 1h and analyzed for release of 6-keto-PGF1alpha, PGE2 and TxB2 by EIA. BDL increased Guinea pig gallbladder cell culture basal PGE2 and PGI2 release which was in part due to increased COX-2 content. CCK incubation down-regulated BDL Guinea pig gallbladder cell culture release of 6-keto-PGF1alpha and PGE2 and down-regulated COX-2 content but did not alter the Control group. The decrease in CCK-mediated BDL cell Guinea pig gallbladder release may be an endogenous mechanism to limit physiologic derangements induced by increased endogenous gallbladder PG synthesis during early acute cholecystitis.
Loss of the short arm of chromosome 8 is a common event in prostatic neoplasms. Previous studies indicate that there may be up to three separate tumor suppressor genes on chromosome arm 8p, based on ...patterns of allelic loss. The responsible gene or genes have yet to be identified. In the present study, we used laser‐capture microdissection of primary human prostate tumors and 17 microsatellite markers across chromosome band 8p21 to determine a minimal deletion interval. From an initial set of 120 cases, three tumors contained overlapping interstitial deletions on chromosome band 8p21. The three cases define an internally consistent minimal candidate tumor suppressor gene interval of approximately two megabases. Published 2002 Wiley‐Liss, Inc.
Two new macromolecular crystallography (MX) beamlines at the National Synchrotron Light Source II, FMX and AMX, opened for general user operation in February 2017 Schneider et al. (2013). J. Phys. ...Conf. Ser. 425, 012003; Fuchs et al. (2014). J. Phys. Conf. Ser. 493, 012021; Fuchs et al. (2016). AIP Conf. Proc. SRI2015, 1741, 030006. FMX, the micro-focusing Frontier MX beamline in sector 17-ID-2 at NSLS-II, covers a 5–30 keV photon energy range and delivers a flux of 4.0 × 1012 photons s-1 at 1 Å into a 1 µm × 1.5 µm to 10 µm × 10 µm (V × H) variable focus, expected to reach 5 × 1012 photons s-1 at final storage-ring current. This flux density surpasses most MX beamlines by nearly two orders of magnitude. The high brightness and microbeam capability of FMX are focused on solving difficult crystallographic challenges. The beamline's flexible design supports a wide range of structure determination methods – serial crystallography on micrometre-sized crystals, raster optimization of diffraction from inhomogeneous crystals, high-resolution data collection from large-unit-cell crystals, room-temperature data collection for crystals that are difficult to freeze and for studying conformational dynamics, and fully automated data collection for sample-screening and ligand-binding studies. FMX's high dose rate reduces data collection times for applications like serial crystallography to minutes rather than hours. With associated sample lifetimes as short as a few milliseconds, new rapid sample-delivery methods have been implemented, such as an ultra-high-speed high-precision piezo scanner goniometer Gao et al. (2018). J. Synchrotron Rad. 25, 1362–1370, new microcrystal-optimized micromesh well sample holders Guo et al. (2018). IUCrJ, 5, 238–246 and highly viscous media injectors Weierstall et al. (2014). Nat. Commun. 5, 3309. Overall, the new beamline pushes the frontier of synchrotron crystallography and enables users to determine structures from difficult-to-crystallize targets like membrane proteins, using previously intractable crystals of a few micrometres in size, and to obtain quality structures from irregular larger crystals.
OBJECTIVETo test the hypothesis that the physicochemical properties of perfluorochemical liquid used in partial liquid ventilation can influence ventilatory requirements, pulmonary mechanics, ...microvascular permeability, and vasoactive mediator release in the abnormal lung.
DESIGNProspective, controlled animal study.
SETTINGResearch laboratory in a university setting.
SUBJECTSMale Sprague-Dawley ratssham and intestinal ischemia/reperfusion injury.
INTERVENTIONSTreatment with perfluorochemical partial liquid ventilation (PLVPP-5 or H-130) or conventional mechanical ventilation (CMV) over 60 mins of superior mesenteric artery occlusion and 60 mins of reperfusion.
MEASUREMENTS AND MAIN RESULTSGas exchange, ventilatory requirements, and pulmonary mechanics were measured in vivo. Subsequently, pulmonary vascular resistance, microvascular permeability, and thromboxane were measured by using the isolated perfused lung preparation. PLV with PP-5 required significantly (p < .05) higher positive end-expiratory pressure resulting in increased mean airway pressures and pulmonary vascular resistance in both sham and intestinal ischemia/reperfusion injured animals compared with those treated with CMV or PLV H-130. PLV PP-5 also resulted in significantly (p < .05) lower respiratory compliance and greater microvascular permeability compared with sham animals. Following intestinal ischemia/reperfusion injury, PLV H-130 treated animals had significantly higher (p < .05) respiratory compliance than those treated with PLV PP-5 and a significantly lower (p < .05) intestinal ischemia/reperfusion-mediated increase in microvascular permeability than those treated with CMV or PLV PP-5. Thromboxane levels were significantly higher (p < .01) in injured animals treated with CMV or PLV PP-5 compared with comparably treated shams, were significantly lower (p < .01) in both PLV groups than CMV, and were further attenuated (p < .01) by PLV H-130 compared with PLV PP-5 animals.
CONCLUSIONWe conclude that PLV with perfluorochemical liquids attenuates pulmonary sequelae resulting from remote organ injury and that the extent of lung protection depends on the physicochemical properties of the perfluorochemical liquids.
This study evaluated the morbidity, mortality, and intermediate term follow-up of patients undergoing replacement of their aortoiliac-femoral systems with lower extremity deep and superficial veins.
...The most commonly used treatment for aortic prosthetic infection is ectopic bypass and removal of the prosthesis. The overall mortality rate with this approach is approximately 20%, with an amputation rate of 10% to 14%. Other limitations include thrombosis of the ectopic bypass leading to limb loss, reinfection of the ectopic bypass, and aortic stump blowout. Dissatisfaction with this approach has led the authors to develop the following.
A neo-aortoiliac system (NAIS) was fashioned from lower extremity deep veins (DV), greater saphenous veins (GSV), or both in patients with infected aortobifemoral prosthesis (n = 17) and other complex aortic problems (n = 3). Removal of infected prosthetic material, harvest of vein, and creation of NAIS was performed as a single-staged procedure.
The in-hospital mortality and amputation rates were 10% each. The mean (+/- standard deviation SD) operative time was 6.5 +/- 1.8 hours and the blood transfusion requirement was 4 +/- 3 units. Four patients experienced postoperative gastrointestinal complications with peritonitis and sepsis; NAIS vein graft resisted infection and remained intact. The mean follow-up time was 22.5 +/- 16 months. NAISs constructed from GSVs were prone to the development of focal stenoses requiring intervention or diffuse neointimal hyperplasia leading to occlusion. In contrast, all NAISs from larger caliber DVs have remained widely patent. The failure rate of GSV NAISs was 64%, compared to 0% for DV NAISs (p = 0.006). Despite the high failure rate in patients with GSV NAISs, none has required amputation. In patients who had DVs harvested for NAIS reconstruction, limb edema and other signs of venous hypertension have been minimal.
NAIS reconstruction from lower extremity veins is a successful option in patients with extensive aortic prosthetic infection and other complex aortic problems.
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