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The application of a Brillouin distributed sensor for the monitoring of railway traffic is presented in this work. The field test is performed on the Italian regional line San Severo-Peschici, operated by Ferrovie del Gargano. A single-mode optical fiber sensor was attached along a rail length of about 60 m. The strain associated with train passage was acquired along the monitored rail length at 31 Hz acquisition rate and 1 m spatial resolution. The data acquired by the sensor demonstrates its capability of retrieving useful information in railway traffic monitoring, such as train identification, axle counting, speed detection, and dynamic load calculation.

In this work, we analyze the effects of Brillouin gain and Brillouin frequency drifts on the accuracy of the differential pulse-width pair Brillouin optical time-domain analysis (DPP-BOTDA). In particular, we demonstrate numerically that the differential gain is highly sensitive to variations in the Brillouin gain and/or Brillouin shift occurring during the acquisition process, especially when operating with a small pulse pair duration difference. We also propose and demonstrate experimentally a method to compensate for these drifts and consequently improve measurement accuracy.

In silica optical fiber Surface Plasmon Resonance (SPR)-based sensors, an increase in fiber core diameter produces a corresponding increase in the sensitivity and Signal to Noise Ratio (SNR). In Plastic Optical Fiber (POF) realized in PMMA there are different influences of design parameters on the performance, as both sensitivity and SNR are concerned. In particular, the SNR, for different refractive index values of the analyte, in a 250 μm diameter POF is greater than the one in 1,000 μm diameter POF. On the other hand, the sensitivity, for the same refractive index values of the analyte, in a 1,000 μm diameter POF is greater than the one in a 250 μm diameter POF. The results of an experimental analysis demonstrating the above behavior are reported.

We propose and demonstrate a long-range Brillouin Optical Time-Domain Analysis (BOTDA) distributed sensing system making use of an unbalanced double sideband probe formed by a Stokes and an anti-Stokes line. In particular, we show that for each measuring condition an optimal Stokes /anti-Stokes input power ratio exists, allowing a larger suppression of nonlocal effects induced by pump depletion. Experiments on a 50 km single-mode sensing fiber with 5 meters spatial resolution are reported., (© 2011 Optical Society of America)

A numerical analysis of conventional and differential pulse-width pair Brillouin optical time domain analysis systems is reported. The tests are focused on determining the performance of these systems especially in terms of spatial resolution, as a function of the pulse characteristics. A new definition of spatial resolution is given, based on analysis of the shape of the Brillouin gain spectrum. The influence of the rise/fall time of the pulse light to the spatial resolution is also studied., (© 2011 Optical Society of America)

Intense nanosecond pulsed electric fields (nsPEFs) interact with cellular membranes and intracellular structures. Investigating how cells respond to nanosecond pulses is essential for a) development of biomedical applications of nsPEFs, including cancer therapy, and b) better understanding of the mechanisms underlying such bioelectrical effects. In this work, we explored relatively mild exposure conditions to provide insight into weak, reversible effects, laying a foundation for a better understanding of the interaction mechanisms and kinetics underlying nsPEF bio-effects. In particular, we report changes in the nucleus of Jurkat cells (human lymphoblastoid T cells) exposed to single pulses of 60 ns duration and 1.0, 1.5 and 2.5 MV/m amplitudes, which do not affect cell growth and viability. A dose-dependent reduction in alkaline comet-assayed DNA migration is observed immediately after nsPEF exposure, accompanied by permeabilization of the plasma membrane (YO-PRO-1 uptake). Comet assay profiles return to normal within 60 minutes after pulse delivery at the highest pulse amplitude tested, indicating that our exposure protocol affects the nucleus, modifying DNA electrophoretic migration patterns.

This paper reports the fabrication and testing of two configurations of optical sensor systems based on Surface Plasmon Resonance (SPR) at the interface of a liquid sample and sandwiched structures realized starting from the exposed core of a Plastic Optical Fiber (POF). The proposed geometries have proven to be suitable for measuring the refractive indexes of liquids whose refractive index falls around 1.35. Furthermore, the proposed sensing head, being low cost and relatively easy to realize, may be very attractive for biosensor implementation.

Intense nanosecond pulsed electric fields (nsPEFs) have been shown to induce, on intracellular structures, interesting effects dependent on electrical exposure conditions (pulse length and amplitude, repetition frequency and number of pulses), which are known in the literature as "bioelectrical effects" (Schoenbach et al., IEEE Trans Plasma Sci 30:293-300, 2002). In particular, pulses with a shorter width than the plasma membrane charging time constant (about 100 ns for mammalian cells) can penetrate the cell and trigger effects such as permeabilization of intracellular membranes, release of Ca(2+) and apoptosis induction. Moreover, the observed effects have led to exploration of medical applications, like the treatment of melanoma tumors (Nuccitelli et al., Biochem Biophys Res Commun 343:351-360, 2006). Pulsed electric fields allowing such effects usually range from several tens to a few hundred nanoseconds in duration and from a few to several tens of megavolts per meter in amplitude (Schoenbach et al., IEEE Trans Diel Elec Insul 14:1088-1109, 2007); however, the biological effects of subnanosecond pulses have been also investigated (Schoenbach et al., IEEE Trans Plasma Sci 36:414-422, 2008). The use of such a large variety of pulse parameters suggests that highly flexible pulse-generating systems, able to deliver wide ranges of pulse durations and amplitudes, are strongly required in order to explore effects and applications related to different exposure conditions. The Blumlein pulse-forming network is an often-employed circuit topology for the generation of high-voltage electric pulses with fixed pulse duration. An innovative modification to the Blumlein circuit has been recently devised which allows generation of pulses with variable amplitude, duration and polarity. Two different modified Blumlein pulse-generating systems are presented in this article, the first based on a coaxial cable configuration, matching microscopic slides as a pulse-delivery system, and the other based on microstrip transmission lines and designed to match cuvettes for the exposure of cell suspensions.

A high-visibility integrated optofluidic Mach-Zehnder interferometer based on liquid-core antiresonant reflecting optical waveguides is reported. The device's geometry has been optimized to minimize the intensity imbalance between the two arms for highly unbalanced Mach-Zehnder configurations. This results in a very compact device with a total length of only 2.5 mm and with required liquid volume of about 0.16 nl. High visibility is demonstrated for two interferometers corresponding to different sensing lengths. The devices have been optically characterized, and the measured interference fringes in the transmitted spectra show good agreement with the theoretical ones.

In a recent article [Opt. Express 17, 17317 (2009)] Kovalev et al. claimed that SBS-induced group index is always negligibly small, regardless of the intensity and bandwidth of pump radiation. In this comment, we show that their conclusions are not valid in any practical conditions in which SBS-based slow light experiments are carried out.

We present a technique for dynamic strain measurements in optical fibers based on the stimulated Brillouin scattering interaction between two counterpropagating optical pulses. The technique allows for a high sampling rate and permits to addressing dynamically and randomly the position at which vibration is measured. Preliminary experimental results carried out with a perturbation frequency up to 98 Hz demonstrate the validity of the proposed technique.

We report on an experimental analysis of stimulated Brillouin scattering (SBS) in a 20-m-long highly birefringent microstructure fiber for sensing applications. In particular, an experimental setup based on Brillouin optical frequency-domain analysis, operating at a wavelength of 1550 nm, has been employed in order to analyze the distribution of Brillouin frequency shift along the fiber, as well as to study the dependence of Brillouin frequency shift on optical polarization, temperature, and strain. Our results indicate that, for any fixed polarization, the fiber has a dual-peaked Brillouin spectrum. A study about the origin of these two peaks is presented.

A Blumlein pulse generator which utilizes the superposition of electrical pulses launched from two individually switched pulse forming lines has been designed and tested. By using a power metal-oxide-semiconductor field-effect transistor as a switch on each end of the Blumlein line, we were able to generate pulses with amplitudes of 1 kV across a 100 Omega load. Pulse duration and polarity can be controlled by the temporal delay in the triggering of the two switches. Using this technique, we have demonstrated the generation of pulses with durations between 8 and 60 ns. The lower limit in pulse duration was determined by the switch closing time and the upper limit by the length of the pulse forming line. A further advantage of the concept is that pulse distortions caused by the non-negligible on-resistance of a line with a single switch can be eliminated by using switches with identical characteristics.

The single-wall carbon nanotubes (SWCNTs) are one of the new materials ofemerging technologies. They are becoming increasingly studied for the possibleapplications in electronics, optics and biology. In particular, very promising fields ofapplication are the development of optical biosensors and the intracellular drug delivery.Nevertheless, there is a paucity of information on their toxicological properties and onpotential human health risk. In the present study the SWCNTs were investigated for thepossible induction of toxicity in human blood cells. Cell growth, viability, apoptosis andmetabolic activity were evaluated in proliferating human peripheral blood lymphocytes. Inun-stimulated human leukocytes primary DNA damage was also evaluated. SWCNTsconcentrations ranging from 1 to 50 μg/ml were tested, and treatment duration varied from6 to 72 h, in accordance with the biological target investigated. A statistically significantdecrease in cell growth was found in cells treated with the highest concentrations (25 and50 μg/ml). Such decrease was not associated to cell death or apoptosis, but it wasdemonstrated to be related to a decrease in metabolic activity, as assessed by resazurinassay. Moreover, treatments of 6 h with SWCNTs concentrations of 1, 5 and 10 μg/mlfailed to induce primary DNA damage on the entire human leukocytes population.

D-Galactose/D-glucose-binding protein from E. coli (GGBP) is a monomer thatbinds glucose with high affinity. The protein structure of GGBP is organized in twoprincipal domains linked by a hinge region that form the sugar-binding site. In this workwe show that the mutant form of GGBP at the amino acid position 182 can be utilized as aprobe for the development of a non-consuming analyte fluorescence biosensor to monitorthe glucose level in diabetes health care.

Starting from the standard three-wave SBS coupled equations, we derive a novel expression describing Brillouin interaction between a pulsed pump wave with a finite cw component, and a Stokes continuous wave counter-propagating along a single-mode optical fiber. The derived integral equation relates the time-domain Stokes beam amplification to the Brillouin frequency distribution. The proposed model permits an accurate description of the Brillouin interaction even for arbitrarily-shaped pump pulses, and can be efficiently employed for improving the accuracy and the resolution of SBS-based distributed sensors. The validity and the limits of the proposed model are numerically analyzed and discussed.

This paper describes an innovative integrated micro flow cytometer that presents a new arrangement for the excitation/detection system. The sample liquid, containing the fluorescent marked particles/cells under analysis, is hydrodynamically squeezed into a narrow stream by two sheath flows so that the particles/cells flow individually through a detection region. The detection of the particles/cells emitted fluorescence is carried out by using a collection fiber placed orthogonally to the flow. The device is based on silicon hollow core antiresonant reflecting optical waveguides (ARROWs). ARROW geometry allows one to use the same channel to guide both the sample stream and the fluorescence excitation light, leading to a simplification of the optical configuration and to an increase of the signal-to-noise ratio. The integrated micro flow cytometer has been characterized by using biological samples marked with standard fluorochromes. The experimental investigation confirms the success of the proposed microdevice in the detection of cells.

Stimulated Brillouin scattering (SBS) has been recently shown to offer a mechanism for generating tunable all-optical delays in room-temperature single-mode optical fibers at telecommunication wavelengths. This technique makes use of the rapid variation of the refractive index that occurs in the vicinity of the Brillouin gain resonance. When the slow light pulse delay is subject to a constraint on the allowable pulse distortion, it has been shown that the use of a pair of closely-spaced Brillouin gain lines can increase the distortion-constrained delay, with respect to the single-line configuration. In this paper, we numerically and experimentally demonstrate that the same experimental apparatus usually employed for generating a Brillouin gain doublet, can also be used for achieving three equally-spaced Brillouin gain resonances, further increasing the distortion-constrained pulse delay.

Recently, metal-clad leaky waveguides (MCLW) have been proposed as highly sensitive single point sensor devices for small-volume refractive index (RI) and fluorescence detection. In this paper, we present a theoretical study of the efficiency of MCLW-based sensors on glass substrate, for fluorescence detection. It is shown that MCLWs can be designed in order to obtain an efficient coupling of fluorescence emission with their leaky modes. This leads to a higher directionality of the fluorescence emission into the glass substrate, when compared to the emission near a pure glass/water interface and surface-plasmon coupled emission (SPCE). Numerical analyses also indicate that collecting the fluorescence emission through a water-immersed microscope objective, may result in a 70-fold enhancement of the detectable signal when compared to conventional fluorescence detection carried out on a glass slide.

In this report we show the purification to homogeneity and a partial characterization of a new odorant-binding protein from Canis familiar (CfOBP) nasal mucosa. In addition, we report preliminary data on the utilization of CfOBP as a probe for the development of a refractive index-based biosensor.

In the last years, fiber optic sensors have been widely exploited for several sensing applications, including static and dynamic strain measurements up to acoustic detection. Among these, fiber Bragg grating sensors have been indicated as the ideal candidate for practical structural health monitoring in light of their unique advantages over conventional sensing devices. Although this class of sensors has been successfully tested for static and low-frequency measurements, the identification of sensor performances for high-frequency detection, including acoustic emission and ultrasonic investigations, is required. To this aim, the analysis of feasibilty on the use of fiber Bragg grating sensors as ultrasonic detectors has been carried out. In particular, the response of fiber Bragg gratings subjected to the longitudinal ultrasonic (US) field has been theoretically and numerically investigated. Ultrasonic field interaction has been modeled, taking into account the direct deformation of the grating pitch combined with changes in local refractive index due to the elasto-optic effect. Numerical results, obtained for both uniform and Gaussian-apodized fiber Bragg gratings, show that the grating spectrum is strongly influenced by the US field in terms of shape and central wavelength. In particular, a key parameter affecting the grating response is the ratio between the US wavelength and the grating length. Normal operation characterized by changes in wavelength of undistorted Bragg peak is possible only for US wavelengths longer than the grating length. For US wavelengths approaching the grating length, the wavelength change is accompanied by subpeaks formation and main peak amplitude modulation. This effect can be attributed to the nonuniformity of the US perturbation along the grating length. At very high US frequencies, the grating is not sensitive any longer. The results of this analysis provide useful tools for the design of grating-based ultrasound sensors for meeting specific requirements in terms of field intensity and frequencies.

A numerical and experimental analysis of the stimulated Brillouin scattering in a single-mode optical fiber for distributed sensing applications is carried out in the frequency domain. The theoretical model describing the Brillouin interaction is solved by taking into account the temporal dynamics of the acoustic wave that is involved. The simulations and the experimental results reveal the role played by the ac component of the acoustic wave, which is responsible for significant changes of the small-signal stimulated Brillouin scattering transfer function that occur when the modulation frequency rises above the natural Brillouin gain spectrum linewidth. One should take these effects into account to perform accurate signal processing of frequency-domain signals in high-resolution distributed sensing applications.

A simple integrated optical refractometric sensor based on hollow-core antiresonant reflecting optical waveguides is proposed. The sensor uses the antiresonant reflecting guidance mechanism and permits one to measure the refractive index of a liquid filling the core by simply monitoring the transmitted spectrum. The device has been made with standard silicon technology, and the experimental results confirm numerical simulations performed in one- and two-dimensional geometry. The sensor exhibits a linear response over a wide measurement range (1.3330-1.4450) and a resolution of 9 x 10(-4) and requires a small analyte volume.

A telomerase assay has been developed for high-throughput screening in 96-well microtiter plates. A crude cell lysate which adds telomere repeats to a biotinylated DNA primer is the source of telomerase. The telomerase-extended primer is hybridized to a digoxigenin-labeled telomere antisense DNA probe. The hybrid is further processed by enzyme-linked immunosorbent assay (ELISA) as follows. The biotinylated hybrid is captured on streptavidin-coated microtiter plates. The immobilized hybrid is probed with alkaline phosphatase-antidigoxigenin and detected via chemiluminescent readout. The limit of detection of a chemically synthesized tetra-telomere repeat was about 10 attomoles. Apparent telomerase activity was detected in lysates of 293T cells. The signal to background for the assay (ratio of signal for the complete assay mixture divided by the signal for the assay mixture without primer) was around 10. An automated system that performed unattended runs of up to 17 96-well microtiter plates in 8h was constructed.

A novel approach to distributed fiber-optic Brillouin sensing is presented and numerically analyzed. An integral equation that directly relates the Brillouin gain to the Brillouin signal is derived in the frequency domain, and from this result a new technique for the quantitative reconstruction of temperature-strain profiles along an optical fiber is developed. We achieve the reconstruction by minimizing a cost function that represents the error between the measured and the model data. We effectively perform such a minimization by representing the unknown (temperature-strain) profile with a finite number of parameters. Numerical results confirm the effectiveness of the proposed approach and its stability against noise in the data.

An integrated optical waveguide refractometer, believed to be novel, is presented. The sensor is based on an antiresonant reflecting optical waveguide and uses the strong attenuation dependence on the refractive index of antiresonant cladding layers as the sensing principle. The theory and the operation of the sensor are discussed in terms of one- and two-dimensional geometry. The theoretical predictions and numerical analysis show that a versatile sensor can be realized. The design trade-offs are discussed, and the sensitivity and measurement range are presented.

The interactions of brain-derived neurotrophic factor (BDNF) with the extracellular domain of its receptor (trkB) are investigated by employing isotope-edited Fourier transform IR (FTIR) spectroscopy. The protein secondary structures of individual BDNF and trkB in solutions are compared with those in their complex. The temperature dependence of the secondary structures of BDNF, trkB, and their complex is also investigated. Consistent with the crystal structure, we observe by FTIR spectroscopy that BDNF in solution contains predominantly beta strands (approximately 53%) and relatively low contents of other secondary structures including beta turns (approximately 16%), disordered structures (approximately 12%), and loops (approximately 18%) and is deficient in alpha helix. We also observe that trkB in solution contains mostly beta strands (52%) and little alpha helix. Conformational changes in both BDNF and trkB are observed upon complex formation. Specifically, upon binding of BDNF, the conformational changes in trkB appear to involve mostly beta turns and disordered structures while the majority of the beta-strand conformation remains unchanged. The IR data indicate that some of the disordered structures in the loop regions are likely converted to beta strands upon complex formation. The FTIR spectral data of BDNF, trkB, and their complex indicate that more amide NH groups of trkB undergo H-D exchange within the complex than those of the ligand-free receptor and that the thermal stability of trkB is decreased slightly upon binding of BDNF. The FT-Raman spectra of BDNF, trkB, and their complex show that the six intramolecular disulfide bonds of trkB undergo significant conformational changes upon binding of BDNF as a result of changes in the tertiary structure of trkB. Taken together, the FTIR and Raman data are consistent with the loosening of the tertiary structure of trkB upon binding of BDNF, which leads to more solvent exposure of the amide NH group and decreased thermal stability of trkB. This finding reveals an intriguing structural property of the neurotrophin ligand-receptor complex that is in contrast to other ligand-receptor complexes such as a cytokine-receptor complex that usually shows protection of the amide NH group and increased thermal stability upon complex formation., (Copyright 2002 John Wiley & Sons, Inc.)

Chordin is a bone morphogenetic protein (BMP) inhibitor that has been identified as a factor dorsalizing the Xenopus embryo. A novel secreted protein, CHL (for chordin-like), with significant homology to chordin, was isolated from mouse bone marrow stromal cells. Injection of CHL RNA into Xenopus embryos induced a secondary axis. Recombinant CHL protein inhibited the BMP4-dependent differentiation of embryonic stem cells in vitro and interacted directly with BMPs, similar to chordin. However, CHL also weakly bound to TGFbetas. In situ hybridization revealed that the mouse CHL gene, located on the X chromosome, was expressed predominantly in mesenchyme-derived cell types: (1) the dermatome and limb bud mesenchyme and, later, the subdermal mesenchyme and the chondrocytes of the developing skeleton during embryogenesis and (2) a layer of fibroblasts/connective tissue cells in the gastrointestinal tract, the thick straight segments of kidney tubules, and the marrow stromal cells in adults. An exception was expression in the neural cells of the olfactory bulb and cerebellum. Interestingly, the spatiotemporal expression patterns of CHL were distinct from those of chordin in many areas examined. Thus, CHL may serve as an important BMP regulator for differentiating mesenchymal cells, especially during skeletogenesis, and for developing specific neurons., (Copyright 2001 Academic Press.)

We have assigned the disulfide structure of Md-65 agouti-related protein (Md65-AGRP) using differential reduction and alkylation followed by direct sequencing analysis. The mature human AGRP is a single polypeptide chain of 112 amino acid residues, consisting of an N-terminal acidic region and a unique C-terminal cysteine-rich domain. The C-terminal domain, a 48 amino acid peptide named Md65-AGRP, was expressed in Escherichia coil cells and refolded under different conditions from the mature recombinant protein. The disulfide bonds in the cystine knot structure of Md65-AGRP were partially reduced using tris(2-carboxyethyl) phosphine (TCEP) under acidic conditions, followed by alkylation with N-ethylmaleimide (NEM). The procedure generated several isoforms with varying degrees of NEM alkylation. The multiple forms of Md65-AGRP generated by partial reduction and NEM modification were then completely reduced and carboxymethylated to identify unreactive disulfide bonds. Differentially labeled Md65-AGRP were directly sequenced and analyzed by MALDI mass spectrometry. The results confirmed that Md65-AGRP contained the same disulfide structure as that of Md5-AGRP reported previously [Bures, E. J., Hui, J. O., Young, Y. et al. (1998) Biochemistry 37, 12172-12177].

Objectives: To examine the distribution of interventional cardiac catheterization laboratories, their case load, the time trends, and the regional variation of percutaneous transluminal cutaneous angioplasty (PTCA) utilization in Italy., Methods: Analysis of data was provided by the annual reports of the Italian Group of Studies and Interventional Cardiology over the period from 1983 to 1996., Results: The number of PTCA facilities and their use steadily increased, mainly in the North. In 1996 the utilization rate was 34 per 100,000 population, but only 60% of labs performed 200 or more procedures., Conclusions: Dramatic time trends and regional variations often took place without an epidemiology and technology assessment-based planning process.

The agouti-related protein gene (Agrp) is a novel gene implicated in the control of feeding behavior. The hypothalamic expression of Agrp is regulated by leptin, and overexpression of Agrp in transgenic animals results in obesity and diabetes. By analogy with the known actions of agouti, these data suggest a role for the Agrp gene product in the regulation of melanocortin receptors expressed in the central nervous system. The availability of recombinant, highly purified protein is required to fully address this potential interaction. A nearly full-length form of AGRP (MKd5-AGRP) was expressed in the cytosolic or soluble fraction of Escherichia coli and appeared as large intermolecular disulfide-bonded aggregates. Following oxidation, refolding, and purification, this protein was soluble, and eluted as a single symmetric peak on RP-HPLC. Circular dichroism studies indicated that the purified protein contains primarily random coil and beta-sheet secondary structure. Sedimentation velocity studies at neutral pH demonstrated that MKd5-AGRP is monomeric at low micromolar concentrations. Mobility shifts observed using SDS-PAGE under reducing and nonreducing conditions for bacterially expressed and mammalian expressed AGRP were identical, an indication of a similar disulfide structure. The purification to homogeneity of a second, truncated form of AGRP (Md65-AGRP) which was expressed in the insoluble or inclusion body fraction is also described. Both forms act as competitive antagonists of alpha-melanocyte stimulating hormone (alpha-MSH) at melanocortin-3 (MC-3) and melanocortin-4 receptors (MC-4). The demonstration that AGRP is an endogenous antagonist with respect to these receptors is a unique mechanism within the central nervous system, and has important implications in the control of feeding.

The agouti-related protein gene (Agrp) plays an important role in body weight regulation. The mature human protein is a single polypeptide chain of 112 amino acid residues, consisting of an N-terminal acidic region and a unique C-terminal cysteine-rich domain. The disulfide structure of recombinant human AGRP was determined by chemical methods using partial reduction with tris(2-carboxyethyl)phosphine under acidic conditions, followed by direct alkylation with N-ethylmaleimide or fluorescein-5-maleimide. Partial reduction and alkylation provided several forms of AGRP that were modified in a stepwise fashion. The resulting proteins were characterized by peptide mapping, sequence analysis, and mass spectrometry, showing that AGRP contained a highly reducible disulfide bond, C85-C109, followed by less reactive ones, C90-C97, C74-C88, C67-C82, and C81-C99, respectively. The chemically defined disulfide connectivity of the recombinant human AGRP was homologous to that of omega-agatoxin IVB except for an additional disulfide bond, C85-C109.

We describe a novel apparatus for the real-time characterization of transverse-mode-mixing effects by monitoring the fluctuations of the M(2) factor defined by Siegman [Proc. Soc. Photo-Opt. Instrum. Eng. 1224, 2 (1990)]. A comparison between the results provided by our approach and those obtained by the use of a standard measurement apparatus has shown a satisfactory agreement within the experimental uncertainty.

A quantitative measurement of laser-beam quality can be performed by determination of the presence of multiple transverse modes of the laser oscillator and by calculation of their power content. Along this line of argument, we discuss a new approach that, starting from near-field and far-field intensity measurements, can evaluate the complex excitation coefficients of the transverse modes in a laser beam. The exploitation of near-field measurements sharply improves the performances of the technique in those cases in which only far-field measurements are used. The validity of the method is confirmed by several accurate numerical simulations and by some experimental results relative to a multimode Q-switched Nd:YAG laser.

Brain-derived neurotrophic factor (BDNF), a 27-kDa noncovalently linked homodimer with subunits of approximately 13.5 kDa as viewed by SDS-PAGE, is thought to be primarily produced in the central nervous system. We report here the isolation of BDNF from pooled normal human sera, using a two-step purification process followed by SDS-PAGE, transfer to a polyvinylidene difluoride membrane, and subsequent identification of the protein by sequence analysis of the appropriate band(s) from the membrane. The level of BDNF in pooled human sera was estimated to be approximately 15 ng/ml as determined by an enzyme-linked immunosorbant assay. The average for six individuals was 18.9 +/- 5.7 ng/ml. There is an approximately 200-fold increase in the levels of BDNF in serum relative to plasma. Results from experiments using differential centrifugation suggest that the source of this increase is due to release from platelets. The presence of high levels of BDNF in serum suggests a role for this neurotrophin either in nerve repair at sites of injured tissue or in nonneuronal functions.

This study was performed to test three instruments for functional status assessment in General Practice: the Dartmouth Coop Charts (COOP Charts), the Functional Status Questionnaire (FSQ) and the Duke University Health Profile (DUHP). All the instruments covered a score of functional aspects in physical, mental and social areas, providing a multidimensional measure of health status. We used these three instruments, validated by international studies, to acquire information concerning their feasibility and acceptability among patients from rural communities needing primary care and to test their validity in differentiating between patient subgroups. The COOP Charts, the FSQ and the DUHP were administered by physicians respectively to 98, 100 and 97 patients, waiting for a visit in the ambulatories of their General Practitioner. Answers relating to each instrument were analyzed according to sex, age and education of patients. All the instruments seemed to be feasible and acceptable, but only the COOP Charts and the FSQ were able to discriminate between different sex, age and scolarity groups. Taking into account the need to elaborate answers according to a formula when using the FSQ, we concluded that the best instrument for General Practice to provide a multidimensional measure of health status seems to be the COOP Charts.

The gene for human stem cell factor (SCF) encodes a leader sequence followed by 248 amino acids (Martin et al., 1990, Cell 63, 203). Of these 248 amino acids, the first 189 correspond to an extracellular domain and the remainder correspond to a hydrophobic transmembrane domain plus a cytoplasmic domain. A naturally occurring soluble form, released by proteolytic cleavage after amino acid 165, has been described. An alternatively spliced mRNA, lacking the codons for exon 6, has also been described. Since the amino acids encoded by exon 6 include the proteolytic cleavage site, the form expressed from the alternatively spliced mRNA tends to remain membrane-bound. In the present study, we have begun to explore structure/function relationships within the extracellular domain of SCF. Forms beginning at amino acid 1 (after the leader sequence) and ranging from 127 to 189 at the C-terminus have been recombinantly expressed in Escherichia coli and purified. In addition, forms missing the amino acids encoded by exon 6, forms missing up to 10 amino acids from the N-terminus, and forms with disulfide bond alterations have been expressed and purified. The forms have been characterized structurally, as well as functionally, in quantitative cell proliferation and receptor-binding assays. The results indicate that amino acids 1-141 comprise a structural and functional core and allow conclusions about the necessity of each of the two disulfide bonds for structure and function.

We have developed a numerical code that, starting from far-field intensity measurements, is able to evaluate the excitation coefficients of the transverse modes in a laser system.Both the coherent and incoherent mode cases are addressed, and, while the incoherent case is shown to be equivalent to a linear problem, the coherent case is discussed through its equivalence to the phase-retrieval problem. Problems arising from both ill posedness and the nonlinearity are discussed in detail.The validity of our approach is confirmed by several numerical simulations and some experimental results on the characterization of a Q-switched Nd:YAG laser.

The phagocyte function of granulocytes and monocytes of whole blood was measured as superoxide production in patients treated for acute nonlymphoblastic leukaemia. The results indicate that the antileukaemic protocol used in this study did not cause a decrease of the oxidative metabolism triggered by phagocytosis and by phorbol esters.

The oxidative metabolism of human neutrophils has been studied after incubation of the cells with recombinant interferon-y. Neutrophils incubated for 2-4 hours with 2-50 U/ml recombinant interferon-y undergo a higher respiratory burst measured both as Oz consumption and Oz- production when stimulated with formyl-methionyl-leucyl-phenylalanine, Concanavalin A or phorbol myristate acetate. The potentiating effect of interferon-y requires more than one hour of incubation, is optimal at 20-50 U/ml and depends on the presence of serum in the incubation medium. The interferon effect depends on new protein synthesis. Cycloheximide at doses which do not alter the respiratory response of normal cells completely prevents the potentiating effect of interferon.

A new technique is successfully used to analyze in real time the pulse-to-pulse fluctuations of mode size and time duration in a picosecond Nd:YAG laser. In particular we show that the pulse length (30 psec) of our active-passive mode-locked Nd:YAG laser is stable to within 10% when the cavityis perfectly tuned and the saturable absorber is fresh. This technique is experimentally shown to be effective and reliable for real-time analysis of the stability of ultrashort laser pulses undera broad range of experimental conditions.

It is widely believed that the transduction pathway in the activation of the NADPH oxidase by formyl-methionyl-leucyl-phenylalanine (FMLP) in neutrophils involves the stimulation of phosphoinositide hydrolysis, the increase in [Ca2+]i and the activity of the Ca2+ and phospholipid dependent protein kinase C. The results presented here show that the activation of the respiratory burst by FMLP can be dissociated by the stimulation of the hydrolysis of phosphatidylinositol 4,5-bisphosphate and Ca2+ changes. In fact, in neutrophils pretreated (primed) with non stimulatory doses of phorbol myristate acetate the respiratory burst by chemotactic peptide is greatly potentiated while the increase in [3H] inositol phosphates formation and in [Ca2+]i are depressed due to the inhibition of phospholipase C. This finding indicates that FMLP can trigger also a sequence of transduction reactions for the activation of the NADPH oxidase different from that involving the formation of the second messengers diacylglycerol and inositol phosphates and the increase in free Ca2+ concentration.

Highly active superoxide (O2-)-forming NADPH oxidase was extracted from plasmamembranes of phorbol-12-myristate-13-acetate-activated pig neutrophils and was partially purified by gel filtration chromatography. Oxidase activity copurified with cytochrome b-245 in an aggregate containing phospholipids and was almost completely separated from FAD and NAD(P)H-cytochrome c reductase. A polypeptide with molecular weight of 31,500 strictly paralleled the purification of NADPH oxidase, suggesting that it is a major component of the enzyme. The enzyme complex was then dissociated by high detergent and salt concentration and cytochrome b-245 was isolated by a further gel filtration chromatography, with a 147 fold purification with respect to the initial preparation. The cytochrome b-245 showed a 31,500 molecular weight by SDS electrophoresis, indicating that it is actually the component previously identified in the partially purified enzyme. The 31,500 protein was phosphorylated in enzyme preparations from activated but not from resting neutrophils, suggesting that phosphorylation of cytochrome b-245 is involved in the activation mechanism of the O2(-) -forming enzyme responsible for the respiratory burst in phagocytes.