Potential of ultraviolet wide-field imaging and multiphoton microscopy for analysis of dehydroergosterol in cellular membranes

Dehydroergosterol (DHE) is an intrinsically fluorescent sterol with absorption/emission in the ultraviolet (UV) region and biophysical properties similar to those of cholesterol. We compared the potential of UV-sensitive low-light-level wide-field (UV-WF) imaging with that of multiphoton (MP) excitation microscopy to monitor DHE in living cells. Significantly reduced photobleaching in MP microscopy of DHE enabled us to acquire three-dimensional z-stacks of DHE-stained cells and to obtain high-resolution maps of DHE in surface ruffles, nanotubes, and the apical membrane of epithelial cells. We found that the lateral resolution of MP microscopy is ～1.5-fold higher than that of UV-WF deconvolution microscopy, allowing for improved spatiotemporal analysis of plasma membrane sterol distribution. Surface intensity patterns of DHE with a diameter of 0.2 μm persisting over several minutes could be resolved by MP time-lapse microscopy. Diffusion coefficients of 0.25-μm-diameter endocytic vesicles containing DHE were determined by MP spatiotemporal image correlation spectroscopy. The requirement of extremely high laser power for visualization of DHE by MP microscopy made this method less potent for multicolor applications with organelle markers like green fluorescent protein-tagged proteins. The signal-to-noise ratio obtainable by UV-WF imaging could be significantly improved by pixelwise bleach rate fitting and calculation of an amplitude image from the decay model and by frame averaging after pixelwise bleaching correction of the image stacks. We conclude that UV-WF imaging and MP microscopy of DHE provide complementary information regarding membrane distribution and intracellular targeting of sterols.     

Congener-specific composition of polychlorinated naphthalenes, coplanar PCBs, dibenzo-p-dioxins, and dibenzofurans in the Halowax series

Concentrations and congener compositions of polychlorinated naphthalenes (PCNs), coplanar polychlorinated biphenyls (Co-PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and dibenzofurans (PCDFs) were determined in seven Halowax (HW) preparations. In HW 1000 and 1031, low-chlorinated naphthalenes (CNs) and in HW 1051, highly chlorinated naphthalenes were dominant, whereas tri- through penta-CNs were major homologues in other Halowaxes. Concentrations of Co-PCBs were in the range of 2.0-2600 ng/g. CB 105 and 118 were dominant in all Halowaxes. Concentrations of PCDDs/DFs were in the range of 92-5900 ng/g. The 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) in Halowaxes calculated from the amounts of PCNs, Co-PCBs, and PCDDs/DFs were in the range of 2800-220,000 ng-TEQ/g. PCNs accounted for most of the total TEQs in Halowaxes, and the contribution of PCDDs/DFs and Co-PCBs to total TEQs was less than 1/1000 that of PCNs. Congeners that most significantly contributed to TEQs were CN 69, 73, 70, and 63. Total TEQs roughly estimated from the Japanese production of technical PCNs, 210 kg-TEQ, was about half from technical PCBs, 440 kg-TEQ on average.     

A new RTD-FET logic family

We describe a new family of clocked logic gates based on the resonant-tunneling diode (RTD). Pairs of RTDs form storage latches, and these are connected by networks consisting of field-effect transistors (FETs), saturated resistors, and RTDs. The design, operation, and expected performance of both a shift register and a matched filter using this logic are discussed. Simulations show that the RTD circuits can achieve higher performance in terms of speed and power in many signal processing applications. Compared to circuits using III-V FETs alone, the RTD circuits are expected to run nearly twice as fast at the same power or at the same speed with reduced power. Compared to circuits using Lincoln Laboratory's fully depleted silicon-on-insulator CMOS, implementation using state-of-the-art RTDs should operate five times faster when both technologies follow the CMOS design rules     

Formation and control of toxic polychlorinated compounds during incineration of wastes containing polychlorinated naphthalenes

To estimate the potential impact on municipal solid waste (MSW) incinerator toxic equivalent (TEQ) emissions of treating wastes containing polychlorinated naphthalenes (PCNs), pilot-scale thermal treatment experiments were conducted. MSW (run 1) and MSW fortified with synthetic rubber belts containing PCNs (runs 2 and 3) were incinerated. Flue-gas and ash samples were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), coplanar polychlorinated biphenyls (co-PCBs), and PCNs. Final exhaust-gas WHO-TEQ emissions were all less than 0.1 ng/Nm3. Flue-gas TEQs were mainly from PCDFs (58-74%). When 2,3,7,8-tetrachlorodibenzo-p-dioxin relative potency factors (REPs) of specific PCN congeners from previous reports were used as estimated toxic equivalency factors to compute estimated PCN TEQs and total TEQs along with PCDDs, PCDFs, and co-PCBs, the contributions of PCNs to the total TEQs were small in ash samples and up to 28% in final exhaust gas. The TEQs in primary combustion flue gases increased through the formation of dioxins and PCNs and then decreased via secondary combustion, fabric filtration, and activated carbon adsorption. From this pilot-scale study, the incremental impact of incinerating PCN-containing wastes on annual TEQ emissions in Japan is estimated as 0.27 g of total TEQ.     

A methodology for the evaluation of research and development projects and associated resource allocation

This paper develops a methodology for the evaluation of research and development projects and the allocation of resources for the development of large scale technologies. The methodology is directed at the problem of selecting a research portfolio when the number of projects is large enough that enumeration of all the possibilities is impractical. In a series of successive screening stages, the number of candidate portfolios is reduced to a practical number. The initial screening stage is based on projected performance of the technology. Successive stages first screen market penetration by ignoring competition from other projects in the portfolio, and then screen by considering project competition. Thus succeeding stages in the methodology are successively finer but more costly to implement. Project screening techniques, which include elimination by aspects, ranking, and stochastic dominance are applied througout the methodology to result in a heuristic and effective approach to evaluation, and associated resource allocation, in very large scale systems.     

Organochlorine contaminants in human adipose tissues from China: mass balance approach for estimating historical Chinese exposure to DDTs

Concentrations of persistent organochlorines (OCs), such as DDTs, hexachlorocyclohexanes (HCHs), hexachlorobenzene (HCB), polychlorinated biphenyls (PCBs), and chlordane compounds (CHLs) were determined in 34 human adipose tissues collected from Guizhou Province, southern China, during 2002. DDT was the predominant contaminant among OCs analyzed; concentrations ranged from 420 to 20 000 ng/g on a lipid wt basis (average +/- sd: 5700 +/- 4100 ng/g). Concentrations of DDTs and the ratio of p,p'-DDT/ sigmaDDT in humans in China were significantly higher than those reported for developed countries. DDT levels in humans in Guizhou Province were comparable to those from Shanghai City, implying the presence of significant sources of DDTs in inland and coastal areas in China. Age-dependent accumulation of HCH concentration was found in this study, possibly because of the considerable reduction in average dairy intake (ADI) of HCHs by Chinese during the recent two decades. On the basis of the information of the ADI rates, half-lives, and body burdens of DDTs, the magnitude of historical DDT exposures by Chinese was estimated. This suggested that approximately 80% of DDTs deposited in Chinese adipose tissues was accumulated before the 1990s. The monitoring of OC levels and epidemiological studies are needed in China to understand the status of contamination and the risks to humans.     

Sterols of the spongeTethya amamensis: Occurrence of (24E)-24-ethylidenecholesta-5,7-dienol, (24E)-24-propylidenecholesta-5,7-dienol,and (24Z)-24-propylidenecholesta-5,7-dienol

The spongeTethya amamensis, collected from Kagoshima Bay, Japan, contained at least 24 sterols, including Δ⁵-sterols (82.2% of total sterols) and Δ^{5, 7}-sterols (17.8%). The predominant sterols were cholesterol (29.0%), cholesta-5,22-dienol (13.8%), 24-methylcholesta-5,22-dienol (10.9%), 24-methylenecholesterol (8.3%), 24-methylcholesta-5,7,22-trienol (6.8%), 24-ethylcholest-5-enol (6.1%), and isofucosterol *4.1%). Combined gas liquid chromatography-mass spectrometry suggested the presence of 3 uncommon sterols, (24E)-24-ethylidenecholesta-5,7-dienol, (24E)-24-propylidenecholesta-5,7-dienol, and (24Z)-24-propylidenecholesta-5,7-dienol as minor components. The sterols ofT. amamensis also contained small amounts of 24-norcholesta-5,7,22-trienol and (24Z)-24-ethylidenecholesta-5,7-dienol.     

Box beams subject to torsion

Stress analysis is carried out for the case where a cantilever box beam is subjected to distributed torque and concentrated torque at the tip. Direct stress $\text{σ}{}_{\mathrm{x}}$ acting in a cover plate is assumed to be in the form of $\text{∑}\text{n}\text{σ}{}_{\mathrm{n}}\text{(x)y}{}^{\mathrm{n}}$ where $\text{n}$ is an odd integer and the fundamental equation governing a box beam is introduced, using variational method. The accuracy can be investigated by comparing the values of strain energy by each obtained solution. Since this calculation is very complicated, strain energy formula is simplified by the new device. From the results of theoretical analysis, it became evident that, for the case where approximation improves, there is no necessity for the empirical formula proposed by Kuhn to be used, and that the range affected by the clamped end is about within the breadth of a box beam from the clamped end.     

Performance Specification of Time-temperature Integrators Designed to Protect Against Botulism in Refrigerated Fresh Foods

ABSTRACT: Time‐temperature integrators (TTIs) may provide a means to monitor safety of fresh foods packaged in reduced‐oxygen environments. Commercially, TTI's have not been fully realized, possibly due to confusion over selection, performance, and reliability. The purpose of this work was to develop a framework for establishing performance targets for TTI's with foods that have potential to cause foodborne botulism. An Arrhenius‐type curve was developed that offers safety margins that satisfy regulatory and shelf life requirements. TTI's developed as specified should minimize “false unsafe” readings that lead to destruction of safe product. The approach could be used to establishing guidelines for application of TTI's to food safety verification.