Ultrafast X-ray probing of water structure below the homogeneous ice nucleation temperature.

Water has a number of anomalous physical properties, and some of these become drastically enhanced on supercooling below the freezing point. Particular interest has focused on thermodynamic response functions that can be described using a normal component and an anomalous component that seems to diverge at about 228 kelvin (refs 1-3). This has prompted debate about conflicting theories that aim to explain many of the anomalous thermodynamic properties of water. One popular theory attributes the divergence to a phase transition between two forms of liquid water occurring in the 'no man's land' that lies below the homogeneous ice nucleation temperature (TH) at approximately 232 kelvin and above about 160 kelvin, and where rapid ice crystallization has prevented any measurements of the bulk liquid phase. In fact, the reliable determination of the structure of liquid water typically requires temperatures above about 250 kelvin. Water crystallization has been inhibited by using nanoconfinement, nanodroplets and association with biomolecules to give liquid samples at temperatures below TH, but such measurements rely on nanoscopic volumes of water where the interaction with the confining surfaces makes the relevance to bulk water unclear. Here we demonstrate that femtosecond X-ray laser pulses can be used to probe the structure of liquid water in micrometre-sized droplets that have been evaporatively cooled below TH. We find experimental evidence for the existence of metastable bulk liquid water down to temperatures of 227(-1)(+2) kelvin in the previously largely unexplored no man's land. We observe a continuous and accelerating increase in structural ordering on supercooling to approximately 229 kelvin, where the number of droplets containing ice crystals increases rapidly. But a few droplets remain liquid for about a millisecond even at this temperature. The hope now is that these observations and our detailed structural data will help identify those theories that best describe and explain the behaviour of water.

Single-particle structure determination by correlations of snapshot X-ray diffraction patterns.

Diffractive imaging with free-electron lasers allows structure determination from ensembles of weakly scattering identical nanoparticles. The ultra-short, ultra-bright X-ray pulses provide snapshots of the randomly oriented particles frozen in time, and terminate before the onset of structural damage. As signal strength diminishes for small particles, the synthesis of a three-dimensional diffraction volume requires simultaneous involvement of all data. Here we report the first application of a three-dimensional spatial frequency correlation analysis to carry out this synthesis from noisy single-particle femtosecond X-ray diffraction patterns of nearly identical samples in random and unknown orientations, collected at the Linac Coherent Light Source. Our demonstration uses unsupported test particles created via aerosol self-assembly, and composed of two polystyrene spheres of equal diameter. The correlation analysis avoids the need for orientation determination entirely. This method may be applied to the structural determination of biological macromolecules in solution.

Femtosecond dark-field imaging with an X-ray free electron laser.

The emergence of femtosecond diffractive imaging with X-ray lasers has enabled pioneering structural studies of isolated particles, such as viruses, at nanometer length scales. However, the issue of missing low frequency data significantly limits the potential of X-ray lasers to reveal sub-nanometer details of micrometer-sized samples. We have developed a new technique of dark-field coherent diffractive imaging to simultaneously overcome the missing data issue and enable us to harness the unique contrast mechanisms available in dark-field microscopy. Images of airborne particulate matter (soot) up to two microns in length were obtained using single-shot diffraction patterns obtained at the Linac Coherent Light Source, four times the size of objects previously imaged in similar experiments. This technique opens the door to femtosecond diffractive imaging of a wide range of micrometer-sized materials that exhibit irreproducible complexity down to the nanoscale, including airborne particulate matter, small cells, bacteria and gold-labeled biological samples.

New light on disordered ensembles: ab initio structure determination of one particle from scattering fluctuations of many copies.

We report on the first experimental ab initio reconstruction of an image of a single particle from fluctuations in the scattering from an ensemble of copies, randomly oriented about an axis. The method is applicable to identical particles frozen in space or time (as by snapshot diffraction from an x-ray free electron laser). These fluctuations enhance information obtainable from an experiment such as conventional small angle x-ray scattering.

Cryptotomography: reconstructing 3D Fourier intensities from randomly oriented single-shot diffraction patterns.

We reconstructed the 3D Fourier intensity distribution of monodisperse prolate nanoparticles using single-shot 2D coherent diffraction patterns collected at DESY's FLASH facility when a bright, coherent, ultrafast x-ray pulse intercepted individual particles of random, unmeasured orientations. This first experimental demonstration of cryptotomography extended the expansion-maximization-compression framework to accommodate unmeasured fluctuations in photon fluence and loss of data due to saturation or background scatter. This work is an important step towards realizing single-shot diffraction imaging of single biomolecules.

Coulomb fission event resolved progeny droplet production from isolated evaporating methanol droplets.

An electrodynamic balance was used to levitate a single methanol droplet while it evaporated down to, and just beyond, its first encountered Coulomb fission limit as ascertained by the time dependence of its laser light scatter signal. At its Coulomb limit, the primary droplet fragmented and approximately 81% of its net charge was released in the form of small droplets that are termed progeny. The window of time over which the progeny droplets were ejected from the electrodynamic balance was deltat = 850 ms, indicating that these droplets were formed with a range of mass-to-charge values. An average of 13.2 (+/-4.4) progeny droplets were detected as a result of this fragmentation event. On the assumption that the average initial radius of the progeny droplets were one-tenth the radius of the droplet undergoing the Coulomb fission event, approximately 50% of the total net charge ejected would not have been carried by the 13 progeny droplets detected.

Antiviral activity of 2'-deoxy-3'-oxa-4'-thiocytidine (BCH-10652) against lamivudine-resistant human immunodeficiency virus type 1 in SCID-hu Thy/Liv mice.

Oral administration of 2'-deoxy-3'-oxa-4'-thiocytidine (BCH-10652), a nucleoside analog structurally similar to lamivudine (3TC), caused dose-dependent inhibition of viral replication in SCID-hu Thy/Liv mice infected with human immunodeficiency virus type 1 NL4-3 and with an NL4-3 clone containing the M184V mutation in reverse transcriptase that confers resistance to 3TC. These experiments demonstrate the utility of this mouse model for evaluating drug resistance and for performing direct comparisons between antiviral compounds in vivo.

Thiosulfate elimination and permeability in a sulfide-adapted marine invertebrate.

Oxidation of hydrogen sulfide to thiosulfate is one of the best-characterized mechanisms by which animals adapted to sulfide minimize its toxicity, but the mechanism of thiosulfate elimination in these animals has remained unclear. In this study, we examined the accumulation and elimination of thiosulfate in the sulfide-adapted marine worm Urechis caupo. The coelomic fluid of U. caupo exposed to 50-100 micromol L-1 sulfide in hypoxic seawater (Po2 ca. 10 kPa) accumulated (mean+/-SD) 132+/-41 micromol L-1 thiosulfate after 2 h, reaching 227+/-113 micromol L-1 after an additional 4 h in aerated, sulfide-free seawater. In whole-animal thiosulfate clearance studies, the rate of thiosulfate elimination from the coelomic fluid followed a single exponential time course with a half-life of 6 h. The thiosulfate permeability coefficient of isolated preparations mounted in diffusion chambers was 7.6x10-5+/-7. 7x10-5 cm s-1 for the hindgut and 5.5x10-7+/-2.7x10-7 cm s-1 for the body wall. These rates were independent of the direction of net efflux (mucosal-to-serosal or serosal-to-mucosal). Using a simple mathematical model of U. caupo that incorporates the thiosulfate permeability coefficients, the thiosulfate half-life was calculated to be 23 h without hindgut ventilation but less than 1 h with normal hindgut ventilation. Based on this information, we propose that passive thiosulfate diffusion across the hindgut is adequate to explain the observed rates of thiosulfate elimination.

CCR5- and CXCR4-utilizing strains of human immunodeficiency virus type 1 exhibit differential tropism and pathogenesis in vivo.

CCR5-utilizing (R5) and CXCR4-utilizing (X4) strains of human immunodeficiency virus type 1 (HIV-1) have been studied intensively in vitro, but the pathologic correlates of such differential tropism in vivo remain incompletely defined. In this study, X4 and R5 strains of HIV-1 were compared for tropism and pathogenesis in SCID-hu Thy/Liv mice, an in vivo model of human thymopoiesis. The X4 strain NL4-3 replicates quickly and extensively in thymocytes in the cortex and medulla, causing significant depletion. In contrast, the R5 strain Ba-L initially infects stromal cells including macrophages in the thymic medulla, without any obvious pathologic consequence. After a period of 3 to 4 weeks, Ba-L infection slowly spreads through the thymocyte populations, occasionally culminating in thymocyte depletion after week 6 of infection. During the entire time of infection, Ba-L did not mutate into variants capable of utilizing CXCR4. Therefore, X4 strains are highly cytopathic after infection of the human thymus. In contrast, infection with R5 strains of HIV-1 can result in a two-phase process in vivo, involving apparently nonpathogenic replication in medullary stromal cells followed by cytopathic replication in thymocytes.

New iridoids from the medicinal plant Barleria prionitis with potent activity against respiratory syncytial virus.

Two new iridoid glycosides (1 and 2), together with the known compounds barlerin (3) and verbascoside (4), were isolated from Barleria prionitis. The new iridoid glycosides were determined to be 6-O-trans-p-coumaroyl-8-O-acetylshanzhiside methyl ester (1) and its cis isomer (2) by using spectroscopic, especially 2D NMR, data. A 3:1 mixture of 1 and 2 was shown to have potent in vitro activity against respiratory syncytial virus (EC50 2.46 microgram/mL, IC50 42.2 microgram/mL).

Percutaneous cholecystolithotomy with endoscopic lithotripsy by using a pulsed-dye laser: preliminary experience.

Percutaneous cholecystolithotomy with endoscopically directed lithotripsy with a pulsed-dye laser was performed in eight patients for the treatment of symptomatic gallstones. All patients had stones that were not amenable to therapy by less invasive techniques. During the first stage of a two-stage procedure, a cholecystostomy catheter was placed percutaneously. During the second stage, performed at least 10 days later, the stones were fragmented with a laser and removed. Fragmentation of the stones was successful in all patients. In six patients, the gallbladder was completely cleared of stones, as assessed with endoscopy and cholecystography. In two patients, residual fragments remained in the gallbladder. No laser-related complications occurred. This preliminary experience suggests that the pulsed-dye laser is safe and effective in endoscopically directed lithotripsy of gallbladder stones.

Percutaneous pulsed-dye laser lithotripsy of gallbladder stones in swine.

Human gallstones were surgically implanted in the gallbladders of 14 pigs. Nine to 16 days later a sheath was successfully placed percutaneously into the gallbladders of 13 animals using ultrasound and fluoroscopy. Two methods were used to guide laser fragmentation: (1) fluoroscopy and a steerable double lumen catheter (two animals), and (2) a flexible endoscope (11 animals). Laser treatment was done in 12 animals with a flashlamp-pumped pulsed-dye laser. A mean of 3600 pulses/animal were delivered using a wavelength of 504 nm and a maximum energy of 60 mJ/pulse. No fragmentation occurred in two animals, partial fragmentation occurred in six, and complete fragmentation occurred in four. Endoscopic guidance was superior to fluoroscopic guidance. Complications (sheath dislodgment, gallbladder perforation, bleeding) occurred in eight of 14 animals. Pulsed-dye laser fragmentation of gallbladder stones is feasible using endoscopic guidance. The use of this technique through an acute percutaneous tract may be associated with complications.

Unequal doses of ioversol versus diatrizoate for urography.

A randomized, double-blind clinical trial was performed to compare a higher dose of conventional ionic contrast media (diatrizoate) with a lower dose of a new, nonionic contrast material (ioversol) for excretory urography (EU). One hundred twenty patients were randomized to receive either 100 ml of diatrizoate (Renografin-60) or 75 ml of ioversol (Optiray-320). Despite the lower iodine dose, the use of ioversol resulted in significantly better opacification of the calyces, renal pelves, and ureters. There was no significant difference in renal parenchymal opacification, opacification of the urinary bladder, or distention of the collecting system. We conclude that high-quality EU can be performed with a lower iodine dose using ioversol.

Needle biopsy of renal allografts: comparison of two techniques.

Two techniques for renal allograft biopsy were retrospectively evaluated to compare relative safety and efficacy. After ultrasound (US) localization of the kidney and biopsy with a hand-held 14-gauge cutting needle, an adequate specimen was obtained in 74 of 77 cases (96%). Major complications occurred in six of these 77 cases (8%). One hundred four biopsies were performed by using a smaller 18-gauge cutting needle with a spring-loaded biopsy "gun" and real-time US guidance. With this newer technique, specimens adequate for diagnosis were obtained in 99 biopsies (95%). There was a single major complication with this technique (1%). The 18-gauge needle with real-time US guidance yields comparably adequate specimens with a lower frequency of complications.

CT evaluation of esophageal perforation following endoscopic sclerotherapy.

We report a case of contained esophageal perforation following endoscopic sclerotherapy for bleeding esophageal varices. CT demonstration of the perforation is presented.

Residue dynamics of acephate and methamidophos in urban dooryard citrus foliage, Pompano Beach, Florida--August-September 1978.

Residues of acephate and its toxic metabolite methamidophos, attributable to the State-Federal program for eradication of the citrus blackfly (CBF) [Aleurocanthus woglumi Ashby] on citrus foliage, were assessed in urban areas in Pompano Beach, Florida. Eighteen dooryard citrus trees were sampled on two line transects, each ca 1.6 km long, along two city streets. The trees were sampled twice monthly for five months, beginning before chemical treatments were applied, continuing through the acephate treatment period, and ending when residues decreased below the limits of detection. Acephate and methamidophos residues, as high as 302.5 ppm and 15.8 ppm, respectively, were detected on leaves within one day after the first of a series of three treatments. Significant conversion of acephate to methamidophose was observed. Of the 143 samples collected, 114 contained measurable residues of both compounds; methamidophos accounted for an average of 19 percent of the total residues. Both compounds degraded rapidly, however, and residues averaged below 1 ppm approximately four weeks after the third treatment in the series. Average foliar half-lives for acephate and methamidophos were 8.93 days (SD = 2.52) and 8.40 days (SD = 2.55), respectively.

Sodium extrusion by the sea-water-acclimated fiddler crab Uca pugilator: comparison with other marine crustacea and marine teleost fish.

1. Measurements of the blood Na concentration and transepithelial electrical potential (T.E.P.) across Uca pugilator acclimated to sea water indicate that Na is maintained out of electrochemical equilibrium with sea water. The resulting net Na influx as well as the sodium gain due to ingestion of the medium must be balanced by extrarenal Na extrusion. 2. The small T.E.P. (-0.7 mV) and the 'transport numbers' of Na and Cl indicate that the permeability to these ions is equivalent. 3. Removal of external K results in a significant stimulation of unidirectional Na efflux that is dependent upon external Na but is not inhibited by ouabain. 4. Transfer of Uca to K and Na-free sea water results in a 54% decline in unidirectional efflux, which is not due to T.E.P. changes. Readdition of 25mM-K stimulates Na efflux much more than can be accounted for by changes in the T.E.P. Readdition of 25mM-Na to potassium-free sea water does not change the Na efflux. 5. The results indicate that Na extrusion by Uca is via a Na/K exchange mechanism which partially inhibits Na/Na exchange. Cessation of Na/K exchange (in K-free sea water) removes this inhibition and allows rapid Na/Na exchange. It is not known whether Na/K and Na/Na exchange are via the same or parallel carrier systems.