Light-scattering studies of packed stationary phases for capillary electrochromatography

Multiply scattered light through turbid media, packed particles, or compressed powders will inherently have a significantly longer optical path length than that of light which is not scattered. The concept of using the multiply scattered light potentially generated in the packed stationary phase of a capillary electrochromatography (CEC) column for enhanced detection as a result of its increased optical path length was examined. Ultraviolet (UV) light at 365 nm or laser light at 635 nm was focused to a small spot onto the packed section of a 3 microns spherisorb ODS1 CEC column (100 microns i.d.). The light was transported inside the capillary, and an image of the multiply scattered light several millimeters along the capillary was collected using a charged-couple device detector. Even if the spot size was less than 100 microns in diameter, evidence of light scattering was observed at a detection spatial off-set distance of 1-2 mm from the illumination point. When the calcium channel blocking drug felodipine was flushed through the column, the light intensity value dropped (increase in absorbance) to a greater degree at a spatial off-set (1.5 mm) than at the illumination point. The greater absorbance values at the spatial off-set were examined experimentally when felodipine was eluted from the column in the CEC mode in 6 min using MeCN/50 mM TRIS (pH 8.0) (80:20, v/v) at an applied voltage of 300 V/cm and an injection time of 2 s at 10 kV. A factor of 8.5 increase in absorbance was observed at a spatial off-set of 1 mm compared to the value obtained at the illumination point. An efficiency value of approximately 234,000 plates m-1 was obtained for this higher felodipine peak. Higher noise values, however, were also observed with this increase in absorbance. Using a spectrophotometer or an open capillary to obtain reference values for optical length, it was possible to estimate the average optical path length of light traveled through the packed stationary phase when transmitted at a spatial off-set. It was concluded that, although an increase in absorbance of 8.5 was observed at a spatial off-set, this most likely arises from the light being "redirected" and scattered in a straightforward fashion along the capillary. It was expected that if substantial multiple scattering did occur inside the packed stationary phase, a significantly larger absorbance increase would be attained. A number of proposals are thus given to explain the relatively low degree of multiple scattering in this stationary phase and suggestions offered on means to attain even higher absorbance increases at a spatial off-set. Additional potential applications are also discussed.     

Fusion-competent vaccines: broad neutralization of primary isolates of HIV

Current recombinant human immunodeficiency virus (HIV) gp120 protein vaccine candidates are unable to elicit antibodies capable of neutralizing infectivity of primary isolates from patients. Here, "fusion-competent" HIV vaccine immunogens were generated that capture the transient envelope-CD4-coreceptor structures that arise during HIV binding and fusion. In a transgenic mouse immunization model, these formaldehyde-fixed whole-cell vaccines elicited antibodies capable of neutralizing infectivity of 23 of 24 primary HIV isolates from diverse geographic locations and genetic clades A to E. Development of these fusion-dependent immunogens may lead to a broadly effective HIV vaccine.     

Anaerobic digestion of space mission wastes.

The technical feasibility of applying leachbed high-solids anaerobic digestion for reduction and stabilization of the organic fraction of solid wastes generated during space missions was investigated. This process has the advantages of not requiring oxygen or high temperature and pressure while producing methane, carbon dioxide, nutrients, and compost as valuable products. Anaerobic biochemical methane potential assays run on several waste feedstocks expected during space missions resulted in ultimate methane yields ranging from 0.23 to 0.30 L g-1 VS added. Modifications for operation of a leachbed anaerobic digestion process in space environments were incorporated into a new design, which included; (1) flooded operation to force leachate through densified feedstock beds; and (2) separation of biogas from leachate in a gas collection reservoir. This mode of operation resulted in stable performance with 85% conversion of a typical space solid waste blend, and a methane yield of 0.3 Lg per g VS added after a retention time of 15 days. These results were reproduced in a full-scale prototype system. A detailed analysis of this process was conducted to design the system sized for a space mission with a six-person crew. Anaerobic digestion compared favorably with other technologies for solid waste stabilization.     

The crying infant

The crying infant is a common presenting complaint and a difficult diagnostic dilemma that may represent the primary manifestation of a serious or even life-threatening condition. Although many children experience an exacerbation of the normal crying tendencies or minor ailments typical of the early months of life, a significant number of infants have underlying pathologic conditions requiring immediate intervention. This article briefly reviews current and past research on this phenomenon and presents differential diagnoses and recommendations for the evaluation and management of the acute crying episode.     

Isolation and characterization of the Saccharomyces cerevisiae DPP1 gene encoding diacylglycerol pyrophosphate phosphatase

Diacylglycerol pyrophosphate (DGPP) is involved in a putative novel lipid signaling pathway. DGPP phosphatase (DGPP phosphohydrolase) is a membrane-associated 34-kDa enzyme from Saccharomyces cerevisiae which catalyzes the dephosphorylation of DGPP to yield phosphatidate (PA) and then catalyzes the dephosphorylation of PA to yield diacylglycerol. Amino acid sequence information derived from DGPP phosphatase was used to identify and isolate the DPP1 (diacylglycerol pyrophosphate phosphatase) gene encoding the enzyme. Multicopy plasmids containing the DPP1 gene directed a 10-fold overexpression of DGPP phosphatase activity in S. cerevisiae. The heterologous expression of the S. cerevisiae DPP1 gene in Sf-9 insect cells resulted in a 500-fold overexpression of DGPP phosphatase activity over that expressed in wild-type S. cerevisiae. DGPP phosphatase possesses a Mg2+-independent PA phosphatase activity, and its expression correlated with the overexpression of DGPP phosphatase activity in S. cerevisiae and in insect cells. DGPP phosphatase was predicted to be an integral membrane protein with six transmembrane-spanning domains. The enzyme contains a novel phosphatase sequence motif found in a superfamily of phosphatases. A dpp1Delta mutant was constructed by deletion of the chromosomal copy of the DPP1 gene. The dpp1Delta mutant was viable and did not exhibit any obvious growth defects. The mutant was devoid of DGPP phosphatase activity and accumulated (4-fold) DGPP. Analysis of the mutant showed that the DPP1 gene was not responsible for all of the Mg2+-independent PA phosphatase activity in S. cerevisiae.     

Is excess intensive care mortality in the United Kingdom concealed by ICU mortality prediction models?

Eleven viruses isolated between 1993 and 1997 from outbreaks of classical swine fever in the neighbouring countries of Slovakia, The Czech Republic and Austria were compared after partial sequencing of the NS5B and E2 genes. Viruses collected from South-Central and West Slovakia were indistinguishable during a period of four years, even when associated with outbreaks of variable severity. Outbreaks that occurred in the Czech Republic in 1996 involved two types of virus, one of which was related to the Slovakian outbreaks, and the other to Austrian outbreaks. The results show that the molecular-genetic approach can reveal epizootiological relationships between outbreaks that would not otherwise be apparent. Furthermore, the relative genetic stability of the classical swine fever virus in the field, means that quite small sequence differences can have epizootiological significance.     

Effects of induced hypertension on transcranial Doppler ultrasound velocities in patients after subarachnoid hemorrhage

BACKGROUND AND PURPOSE: Transcranial doppler ultrasound (TCD) is used after subarachnoid hemorrhage to detect cerebral vasospasm and is often treated with induced hypertension. Cerebral autoregulation, however, may be disturbed in this population, raising the possibility that TCD velocities may be elevated by induced hypertension. To study this possibility, we performed continuous TCD monitoring of the middle cerebral artery during the induction and withdrawal of induced hypertension in patients after subarachnoid hemorrhage. METHODS: Twenty-eight patients were studied during the induction and withdrawal of hypertension using primarily phenylephrine. Continuous monitoring was performed on the middle cerebral artery with the highest flow velocity. Treatment was based on rising TCD velocities or clinical evidence for cerebral vasospasm. Mean arterial pressure and mean TCD velocities were recorded every minute. A change of > 15% from starting TCD values was considered significant. Cerebral autoregulation was calculated as a percentage of intact autoregulation. Patients were subsequently divided into groups of disturbed and intact autoregulation. RESULTS: In 10 of 19 patients (53%), TCD velocities changed by > 15% and paralleled changes in mean arterial pressure. This directly altered the TCD interpretation of the grade of vasospasm in 7 of 19 patients (36%). Three additional patients had smaller absolute changes in TCD velocities. No clinical difference could be identified between patients with disturbed and intact autoregulation. CONCLUSIONS: In patients with disturbed autoregulation after subarachnoid hemorrhage, induced hypertension can alter cerebral blood flow velocities. The level of autoregulation needs to be considered when interpreting TCD velocities in patients after subarachnoid hemorrhage.