Quantitative methods for scoring cell migration and invasion in filter-based assays

The ability of cells to traverse pores in a biocompatible filter provides means for examining cell chemoattraction. Filter-based assays also permit rapid, quantitative assessment of the in vitro migratory and invasive potential of tumor cells. Scoring migration has relied on visual counting of stained cells which appear on the underside of the filter and determining a true percentage score involves arduous counting of cells on both filter surfaces. Visual counting of random fields may be unreliable, and counting all fields is laborious. In the present study we developed and compared two alternative methods for scoring cell numbers in filter-based assays, a colorimetric assay of toluidine blue binding, and a radioassay of cells prelabeled with [3H]thymidine. Each method was evaluated for sensitivity, variability, ease of use and efficiency, and suitability for use in assays of cell migration and invasion. The radiolabeling method proved to be sensitive and reliable and was the most efficient technique. Although less sensitive and specific, the colorimetric dye method offered a rapid and reliable, nonradioactive alternative with the distinct advantage of preserving intact cultures for follow-up visual assessments. We conclude that colorimetric and radiolabel scoring of filter-based assays are reliable and efficient semiautomated methods which provide means to obtain more complete assessments of cell migration and invasion.     

The neuropeptide red pigment concentrating hormone affects rhythmic pattern generation at multiple sites

The level of LamB protein in the outer membrane of Escherichia coli was derepressed in the absence of a known inducer (maltodextrins) under carbohydrate-limiting conditions in chemostats. LamB protein contributed to the ability of the bacteria to remove sugar from glucose-limited chemostats, and well-characterized lamB mutants with reduced stability constants for glucose were less growth competitive under glucose limitation than those with wild-type affinity. In turn, wild-type bacteria were less growth competitive than lamB mutants with enhanced sugar affinity. In contrast to an earlier report, we found that LamB- bacteria were less able to compete in carbohydrate-limited chemostats (with glucose, lactose, arabinose, or glycerol as the carbon and energy sources) when mixed with LamB+ bacteria. The transport Km for [14C]glucose was affected by the presence or affinity of LamB, but only in chemostat-grown bacteria, with their elevated LamB levels. The pattern of expression of LamB and the advantage it confers for growth on low concentrations of carbohydrates are consistent with a wider role in sugar permeation than simply maltosaccharide transport, and hence the well-known maltoporin activity of LamB is but one facet of its role as the general glycoporin of E. coli. A corollary of these findings is that OmpF/OmpC porins, present at high levels in carbon-limited bacteria, do not provide sufficient permeability to sugars or even glycerol to support high growth rates at low concentrations. Hence, the sugar-binding site of LamB protein is an important contributor to the permeability of the outer membrane to carbohydrates in habitats with low extracellular nutrient concentrations.     

Effects of glucose load on brain extracellular lactate concentration in conscious rats using a microdialysis technique

Changes in the brain lactate concentration in cerebral extracellular fluid (ECF) during intravenous infusion of glucose and local administration of glucose were investigated in adult, conscious, unrestrained rats, with a microdialysis probe in the posterior hippocampus. The rats were infused intravenously with either 25% sucrose solution or 25% glucose solution at a rate of 16.6 microliters.min-1.100 g-1 for three hours. The blood glucose concentration reached 17.0 +/- 2.6 mM at the end of the glucose infusion, and brain ECF glucose showed a parallel change with the blood glucose concentration and increased to 2.37 +/- 0.30 mM. However, blood and brain ECF glucose concentrations did not change in animals infused with the sucrose solution. On the other hand, the blood lactate concentration in the glucose-infused group also increased from 0.93 +/- 0.18 mM to 2.85 +/- 0.39 mM at the end of the glucose infusion, which was significantly higher than that measured in the sucrose-infused group. The blood lactate level in the glucose-infused group returned to the basal level by the end of the experiment. Brain ECF lactate concentrations increased from 1.21 +/- 0.06 mM to 1.69 +/- 0.11 mM in glucose-infused animals, but did not change in the sucrose-infused animals. The brain ECF lactate concentration showed a positive correlation with the brain ECF glucose concentration in glucose-infused animals. Another group of rats was administered glucose locally for 90 min after substitution of artificial cerebrospinal fluid.(ABSTRACT TRUNCATED AT 250 WORDS)     

A selective CVD tungsten-strapped polysilicon local interconnectiontechnology

A local interconnection technology utilizing polysilicon strapped with selective-chemical-vapor-deposited (CVD) tungsten has been developed. Both n- and p-channel MOS transistors have been successfully fabricated using this technology. Tungsten deposited on polysilicon is an attractive gate shunt and local interconnection material because of its low resistivity, immunity to dopant segregation and diffusion, and resistance to electromigration. A potential problem of this technology is the excessive diode leakage current associated with strapping shallow source/drain diodes with tungsten. The leakage is attributed to defects induced by the heavy source/drain implant, which can be effectively eliminated with a proper annealing procedure     

Heading towards an advanced signalling system for multimedia,multiparty services in broadband ISDN

Introduction of the broadband integrated services digital network (B-ISDN) will enable network users to make use of advanced multimedia, multiparty services. With this aim in view, RACE II project R2044, MAGIC, is investigating long-term signalling requirements, architecture and protocols for B-ISDN. New service concepts are being developed in MAGIC which will provide more flexibility when compared to narrowband ISDN (N-ISDN). The service requirements and other signalling requirements, such as the separation between call and associated connections at the network-node interface, are used to determine the distribution of functions over the network. The signalling protocols which are used to exchange the signalling information are subsequently developed. These signalling protocols are simulated to prove the performance and the feasibility of call handling concepts     

All-optical wavelength conversion by semiconductor opticalamplifiers

Following a brief introduction to the applications for wavelength conversion and the different available conversion techniques, the paper gives an in depth analysis of cross gain and cross phase wavelength conversion in semiconductor optical amplifiers. The influence of saturation filtering on the bandwidth of the converters is explained and conditions for conversion at 20 Gb/s or more are identified. The cross gain modulation scheme shows extinction ratio degradation for conversion to longer wavelengths. This can be overcome using cross phase modulation in semiconductor optical amplifiers that are integrated into interferometric structures. The first results for monolithic integrated interferometric wavelength converters are reviewed, and the quality of the converted signals is demonstrated by transmission of 10 Gb/s converted signals over 60 km of nondispersion shifted single mode fiber     

Patient's self-interpretation of tuberculin skin tests

BACKGROUND: In the outpatient use of tuberculin skin testing (purified protein derivative [PPD]), it is at times inconvenient to have a patient revisit for interpretation. Therefore, we assessed patients' ability to self-interpret these test results. METHODS: In keeping with prior custom, patients were seen by an experienced nurse, who performed skin testing with PPD intermediate strength as well as mumps and Candida anergy control tests in some cases, and explained the procedure. The patients were asked to return 48 to 72 h later, at which time one of the researchers recorded their test interpretations before they were again evaluated by the nurse. RESULTS: Sixty-eight patients were studied, of whom 59 returned at appropriate interval. Eighteen patients had a positive PPD test reaction of 10 to 20 mm induration, which only one patient correctly identified as a positive test result. However, positive anergy control tests were correctly interpreted in 10 of 27 cases. CONCLUSION: The small number of positive PPD test result recognition by these patients may be partially attributed to their lack of education, as well as foreign birth and denial of illness. PPD results should be checked by an experienced professional.     

Scattering from coated targets using a frequency-dependent, surfaceimpedance boundary condition in FDTD

One method for reducing the radar cross section of objects such as aircraft and missiles is the application of a lossy coating. Computing scattering from targets coated with dielectric/magnetic materials is challenging due to the reduced wavelengths of an incident field inside the coating. These smaller wavelengths require finer sampling of the fields. A technique for implementing this calculation without greatly increased memory requirements or computation times has previously been developed using a finite-difference time-domain (FDTD) code which has been tested in one, two, and three dimensions. The method requires knowledge of the frequency behaviour of the complex permittivity and permeability, and the thickness of the dielectric coating and is applicable to thin coatings when one or more reflections from the conducting surface are significant. The impedance at the surface of the coating is computed based on the given information and then approximated using a summation of causal functions. The approximated impedance is Z-transformed and added to the FDTD code in special update equations for the fields at the surface of the coating. No computations are required inside the coatings so the FDTD grid can be sized based on the free-space wavelength. The result obtained is valid over the entire frequency range of interest, assuming that the approximated surface impedance is a good match over the entire range. Comparisons with measurements of a scale model coated missile show good agreement and almost no increase in resource requirements over a standard FDTD calculation for an uncoated metal target     

Distributed channel assignment strategies using coexistenceetiquettes for land based radio environment

The performance of two distributed channel assignment algorithms which use coexistence etiquettes for point to point links are considered by the authors. It has been found that selecting channels prior to transmission at the transmitter as opposed to the receiver, supports greater offered traffic with the additional advantage of a less complex protocol