In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate

We report the implementation of an electrostatic Einzel lens (Boersch) phase plate in a prototype transmission electron microscope dedicated to aberration-corrected cryo-EM. The combination of phase plate, C_s corrector and Diffraction Magnification Unit (DMU) as a new electron-optical element ensures minimal information loss due to obstruction by the phase plate and enables in-focus phase contrast imaging of large macromolecular assemblies. As no defocussing is necessary and the spherical aberration is corrected, maximal, non-oscillating phase contrast transfer can be achieved up to the information limit of the instrument. A microchip produced by a scalable micro-fabrication process has 10 phase plates, which are positioned in a conjugate, magnified diffraction plane generated by the DMU. Phase plates remained fully functional for weeks or months. The large distance between phase plate and the cryo sample permits the use of an effective anti-contaminator, resulting in ice contamination rates of <0.6 nm/h at the specimen. Maximal in-focus phase contrast was obtained by applying voltages between 80 and 700 mV to the phase plate electrode. The phase plate allows for in-focus imaging of biological objects with a signal-to-noise of 5-10 at a resolution of 2-3 nm, as demonstrated for frozen-hydrated virus particles and purple membrane at liquid-nitrogen temperature.     

Resting respiratory tract dendritic cells preferentially stimulate T helper cell type 2 (Th2) responses and require obligatory cytokine signals for induction of Th1 immunity

Consistent with their role in host defense, mature dendritic cells (DCs) from central lymphoid organs preferentially prime for T helper cell type 1 (Th1)-polarized immunity. However, the "default" T helper response at mucosal surfaces demonstrates Th2 polarity, which is reflected in the cytokine profiles of activated T cells from mucosal lymph nodes. This study on rat respiratory tract DCs (RTDCs) provides an explanation for this paradox. We demonstrate that freshly isolated RTDCs are functionally immature as defined in vitro, being surface major histocompatibility complex (MHC) II lo, endocytosishi, and mixed lymphocyte reactionlo, and these cells produce mRNA encoding interleukin (IL)-10. After ovalbumin (OVA)-pulsing and adoptive transfer, freshly isolated RTDCs preferentially stimulated Th2-dependent OVA-specific immunoglobulin (Ig)G1 responses, and antigen-stimulated splenocytes from recipient animals produced IL-4 in vitro. However, preculture with granulocyte/macrophage colony stimulating factor increased their in vivo IgG priming capacity by 2-3 logs, inducing production of both Th1- and Th2-dependent IgG subclasses and high levels of IFN-gamma by antigen-stimulated splenocytes. Associated phenotypic changes included upregulation of surface MHC II and B7 expression and IL-12 p35 mRNA, and downregulation of endocytosis, MHC II processing- associated genes, and IL-10 mRNA expression. Full expression of IL-12 p40 required additional signals, such as tumor necrosis factor alpha or CD40 ligand. These results suggest that the observed Th2 polarity of the resting mucosal immune system may be an inherent property of the resident DC population, and furthermore that mobilization of Th1 immunity relies absolutely on the provision of appropriate microenvironmental costimuli.     

Equaliser Performance for HIPERLAN in indoor channels

In order to accomplish practical deployment modelling for system performance evaluation and comparison for possible modulation and equalisation schemes to be used in HIPERLAN, a wide band tapped delay line (WTDL) channel model has been adopted by ETSI to characterise the multipath fading in the indoor radio environment. Based on this statistical channel model, and using Monte Carlo method, this paper evaluates the average probability of error for linear and decision feedback equaliser as a function of signal-to-noise ratio. It also evaluates the matched filter bound for this channel model. The results show the optimum performance levels achievable via the use of any equaliser.The work described in this paper was supported by the UK DTI/EPSRC LINK project: PC2011 High Throughput Radio Modem under EPSRC grant reference GR/K00318 in collaboration with Symbionics Networks limited.     

Electromagnetic-field calculations for irregularly shaped, axisymmetric layered particles with focused illumination

Theoretical procedures are presented for the determination of the internal and the near-surface electromagnetic fields for an arbitrary monochromatic field (e.g., a focused laser beam) incident upon an irregulary shaped, axisymmetric layered particle. The layered spherical particle solution is also given as a special case of the general solution. Systematic calculations are presented that demonstrate the effects of particle shape and incident focused-beam orientation on the electromagnetic-field distributions.     

Chemotherapy in non-small cell lung cancer: a review

Lung cancer, of which non-small cell carcinoma is the most common, has been a significant therapeutic challenge for decades and will remain so for decades to come. Despite its prevalence, progress in the management of non-small cell lung cancer has been relatively slow. This is in part due to the pessimism of most physicians treating this disease, which has resulted in a relatively lackadaisical attitude with regards to clinical trials when compared to other solid tumours like breast or colorectal cancers. Nevertheless, the past decade has seen significant progress, specifically with regards to the management of locally advanced disease. Chemotherapy, though shown to be biologically active in non-small cell lung cancer, is considered an ineffective palliative tool in the setting of metastatic disease due to its toxicities and the "less than encouraging" response rates generated by the cisplatin-based combination regimen which is generally considered to be the most active currently available. The advent of new active agents such as paclitaxel and vinorelbine which are potentially less toxic may change this view. Conversely, the response rate of locally advanced disease to chemotherapy is significantly higher and this has resulted in numerous multimodality trials of neoadjuvant chemotherapy prior to surgery and/or radiation. To date, a number of randomised trials have shown that this approach can result in significant survival benefit for patients with locally advanced disease. An alternative approach makes use of the potential synergism between certain chemotherapeutic agents (such as cisplatin) and radiation when used concurrently. However, data on concurrent chemoradiotherapy in locally advanced disease have been largely based on single-arm studies and are inconclusive. Three randomised trials on concurrent chemoradiotherapy have been shown benefit for the use of combined modality in locally advanced disease. Hence, treatment of locally advanced disease should include chemotherapy as part of the combined modality approach. However, the optimal sequencing of these modalities would require well-designed randomised trials to determine.     

Paging tradeoffs in distributed-shared-memory multiprocessors

Massively parallel processors have begun using commodity operating systems that support demand-paged virtual memory. To evaluate the utility of virtual memory, we measured the behavior of seven shared-memory parallel application programs on a simulated distributed-shared-memory machine. Our results (1) confirm the importance of gang CPU scheduling, (2) show that a page-faulting processor should spin rather than invoke a parallel context switch, (3) show that our parallel programs frequently touch most of their data, and (4) indicate that memory, not just CPUs, must be gang scheduled. Overall, our experiments demonstrate that demand paging has limited value on current parallel machines because of the applications' synchronization and memory reference patterns and the machines' high page-fault and parallel context-switch overheads.An earlier version of this paper was presented at Supercomputing '94.This work is supported in part by NSF Presidential Young Investigator Award CCR-9157366; NSF Grants MIP-9225097, CCR-9100968, and CDA-9024618; Office of Naval Research Grant N00014-89-J-1222; Department of Energy Grant DE-FG02-93ER25176; and donations from Thinking Machines Corporation, Xerox Corporation, and Digital Equipment Corporation.     

A rapid computational filter for predicting the rate of human renal clearance

In silico models that predict the rate of human renal clearance for a diverse set of drugs, that exhibit both active secretion and net re-absorption, have been produced using three statistical approaches. Partial Least Squares (PLS) and Random Forests (RF) have been used to produce continuous models whereas Classification And Regression Trees (CART) has only been used for a classification model. The best models generated from either PLS or RF produce significant models that can predict acids/zwitterions, bases and neutrals with approximate average fold errors of 3, 3 and 4, respectively, for an independent test set that covers oral drug-like property space. These models contain additional information on top of any influence arising from plasma protein binding on the rate of renal clearance. Classification And Regression Trees (CART) has been used to generate a classification tree leading to a simple set of Renal Clearance Rules (RCR) that can be applied to man. The rules are influenced by lipophilicity and ion class and can correctly predict 60% of an independent test set. These percentages increase to 71% and 79% for drugs with renal clearances of < 0.1 ml/min/kg and > 1 ml/min/kg, respectively. As far as the authors are aware these are the first set of models to appear in the literature that predict the rate of human renal clearance and can be used to manipulate molecular properties leading to new drugs that are less likely to fail due to renal clearance.     

Walk-up VR: virtual reality beyond projection screens

Today, VR research and development efforts often focus on the continual innovation of interaction styles and metaphors for virtual environments (VEs). New tools and interaction devices aim to increase the immersive experience rather than support seamless integration with real work scenarios. Even though users may soon perceive odours within VEs, real, task-oriented interaction within these environments will continue to lag behind. Combining these efforts can result in new user interfaces that reduce the cumbersome barriers prevalent in VEs today, finally unleashing the latent impact of this technology in everyday life. Implementing this vision requires an interdisciplinary and applied approach to integrate VR into the workplace. Mixed-reality display capabilities, useful multimodal interaction and perceptual, intuitive interfaces are major components of such an application-oriented and human-centered approach, for which we coined the term walk-up VR. The paper discusses augmented and virtual reality as contributing technologies     

Radiopharmacology of inhaled 133Xe in skeletal sites containing deposits of Gaucher cells

Gaucher's disease is a lysosomal storage disease in which cells of the reticuloendothelial system accumulate the lipid glucocerebroside. It is characterized by slowly progressive visceral and osseous involvement. One of the latter manifestations includes lipid infiltration of bone marrow. We monitored the rate of inhaled 133Xe uptake and wash-out over diseased and normal metaphyseal and epiphyseal areas of the knee. Twenty-two patients (15 adults, 7 children) with various degrees of previously diagnosed Gaucher's disease were positioned supine under a gamma-camera interfaced to a computer system. All patients rebreathed 133Xe gas from a closed system for 10 min followed by 14 min of wash-out. Digitized images of the lung, liver, spleen, bony sites and soft tissue were obtained at 1 min intervals during the wash-in and wash-out phases. Counts for each ROI were normalized per 100 pixels and plotted as a function (time). Maximum uptake was also calculated by relating the counts/ROI/100 pixels to the 10 min integrated lung count during equilibrium (the administered "dose"). There was essentially no 133Xe uptake in liver and spleen involved with Gaucher's disease. Monophasic uptake and biphasic wash-out curves were observed in the limited investigative population. Skeletal Gaucher deposits released the 133Xe at a greater rate relative to soft tissue.     

A study of the polymerization behaviour of N-(4-phenoxy)-phenylmaleimide using DSC analysis

Summary The thermal cure of a monofunctional maleimide N-(4-phenoxy)-phenylmaleimid (1) was examined using differential scanning calorimetry (DSC). The analogous bis-maleimide bis-(4-maleimidophenyl)ether (2) was also analyzed in the same manner for comparison. Polymerization kinetics derived from scanning DSC measurements showed an order of reaction varying between n=1 and 2 for (1), compared to an order of reaction between n=2 and 3 (by multiple linear regression analysis) for (2). The overall enthalpy of polymerization (H_o) for N-(4-phenoxy)-phenylmaleimide (94kJ/mole) agrees well with that obtained for maleimide itself (89kJ/mole) using conventional calorimetric techniques indicating that polymerization appears complete within the timescale of the experiment. In contrast the analogous BMI displays a markedly lower overall polymerization enthalpy (ca. 66kJ/mole) indicating an apparent conversion of only 37%.