Determination of Hydrogen Density by Swift Heavy Ions.

A novel method to determine the total hydrogen density and, accordingly, a precise plasma temperature in a lowly ionized hydrogen plasma is described. The key to the method is to analyze the energy loss of swift heavy ions interacting with the respective bound and free electrons of the plasma. A slowly developing and lowly ionized hydrogen theta-pinch plasma is prepared. A Boltzmann plot of the hydrogen Balmer series and the Stark broadening of the H_{ß} line preliminarily defines the plasma with a free electron density of (1.9±0.1)×10^{16} cm^{-3} and a free electron temperature of 0.8-1.3 eV. The temperature uncertainty results in a wide hydrogen density, ranging from 2.3×10^{16} to 7.8×10^{18} cm^{-3}. A 108 MHz pulsed beam of ^{48}Ca^{10+} with a velocity of 3.652 MeV/u is used as a probe to measure the total energy loss of the beam ions. Subtracting the calculated energy loss due to free electrons, the energy loss due to bound electrons is obtained, which linearly depends on the bound electron density. The total hydrogen density is thus determined as (1.9±0.7)×10^{17} cm^{-3}, and the free electron temperature can be precisely derived as 1.01±0.04 eV. This method should prove useful in many studies, e.g., inertial confinement fusion or warm dense matter.

Excimer laser pumped by an intense, high-energy heavy-ion beam.

High-energy heavy ions are an ideal tool to generate homogeneously excited, extended volumes of nonthermal plasmas. Here, the high-energy loss (dE/dx) and absolute power deposition of heavy ions interacting with matter has been used to pump an ultraviolet laser. A pulsed 70 MeV/u 238U beam with up to 2.5 x 10(9) particles in approximately 100 ns beam bunches was stopped in a 1.2 m long laser cell filled with a 1.6 bar Ar-Kr-F2 mixture (typically 50%:49.9%:0.1%). Laser effect on the 248 nm KrF* excimer transition is clearly demonstrated.

Single photon ionization time-of-flight mass spectrometry with a pulsed electron beam pumped excimer VUV lamp for on-line gas analysis: setup and first results on cigarette smoke and human breath.

Single-photon ionization (SPI) using vacuum ultraviolet (VUV) light produced by an electron beam pumped rare gas excimer source has been coupled to a compact and mobile time-of-flight mass spectrometer (TOFMS). The novel device enables real-time on-line monitoring of organic trace substances in complex gaseous matrixes down to the ppb range. The pulsed VUV radiation of the light source is employed for SPI in the ion source of the TOFMS. Ion extraction is also carried out in a pulsed mode with a short time delay with respect to ionization. The experimental setup of the interface VUV light source/time-of-flight mass spectrometer is described, and the novel SPI-TOFMS system is characterized by means of standard calibration gases. Limits of detection down to 50 ppb for aliphatic and aromatic hydrocarbons were achieved. First on-line applications comprised real-time measurements of aromatic and aliphatic trace compounds in mainstream cigarette smoke, which represents a highly dynamic fluctuating gaseous matrix. Time resolution was sufficient to monitor the smoking process on a puff-by-puff resolved basis. Furthermore, human breath analysis has been carried out to detect differences in the breath of a smoker and a nonsmoker, respectively. Several well-known biomarkers for smoke could be identified in the smoker's breath. The possibility for even shorter measurement times while maintaining the achieved sensitivity makes this new device a promising tool for on-line analysis of organic trace compounds in process gases or biological systems.

Single-photon ionization quadrupole mass spectrometry with an electron beam pumped excimer light source.

The application of soft ionization methods for mass spectrometry (MS), such as single-photon ionization (SPI) using vacuum ultraviolet (VUV) light, provides powerful analytical instrumentation for real-time on-line monitoring of organic substances in gaseous matrixes. A compact and mobile quadrupole mass spectrometer (QMS) system using a novel electron beam pumped rare gas VUV lamp for SPI has been developed for on-line analysis of organic trace compounds (ppb concentrations). The VUV radiation of the light source is employed for SPI in the ion source of the QMS. The concept of the interfacing of the VUV light source with the QMS is described and the SPI-QMS is characterized. On-line detection limits down to 50 ppb for benzene, toluene, and m-xylene were achieved. The instrument is well suited for continuous measurements of aromatic and aliphatic trace compounds and can therefore be used for on-line monitoring of trace compounds in dynamically fluctuating process gases. First measurements of gas standards, petrochemical samples, and on-line monitoring of automotive exhaust are presented.

Energy-transfer processes in neon-hydrogen mixtures excited by electron beams.

Energy- and charge-transfer processes in neon-hydrogen mixtures (500-1400 hPa neon and 0.001-3 hPa hydrogen partial pressures) excited by a pulsed low-energy (approximately 10 keV) electron beam were investigated using time-resolved spectroscopy. Time spectra of the hydrogen Lyman-alpha line, neon excimer emission (second continuum), and neon atomic lines (3p-3s transitions) were recorded. The time-integrated intensity of the Lyman-alpha emission was measured for the same range of gas mixtures. It is shown that direct energy transfer from Ne*2 excimers and neon atoms in the four lowest excited states as well as recombination of H3+ ions are the main channels populating atomic hydrogen in the n=2 state. A rate constant of (4.2+/-1.4)x10(-11) cm3 s(-1) was obtained for the charge transfer from Ne2+ ions to molecular hydrogen. A lower limit for the depopulation rate constant of Ne*2 excimers by molecular hydrogen (combination of energy transfer and ionization) was found to be 1.0 x 10(-10) cm3 s(-1).

Single photon ionization (SPI) via incoherent VUV-excimer light: robust and compact time-of-flight mass spectrometer for on-line, real-time process gas analysis.

Fast on-line detection of organic compounds from complex mixtures, such as industrial process gas streams, require selective and sensitive analytical methods. One feasible approach for this purpose is the use of mass spectrometry (MS) with a selective and soft (fragment-free) ionization technique, such as chemical ionization (CI) or photo ionization (PI). Single photon ionization (SPI) with vacuum ultraviolet (VUV) light is a particularly sof tionization technique, well-suited for detection of both aromatic and aliphatic species. Problematic, however, is the generation of the VUV light. In general, the vacuum ultraviolet (VUV) light sources for SPI-MS are based either on lasers (e.g., 118-nm radiation generated by frequency-tripling of the third harmonic of a Nd:YAG laser) or on conventional VUV lamps, such as deuterium lamps. Althoughthe laser-based techniques are very sophisticated and expensive, the conventional lamps have serious drawbacks regarding their optical parameters, such as low-output power, low spectral power density, and broad emission bands. In this work, a novel excimer VUV light source, in which an electron beam is used to form rare gas excimer species, is used. The excimer VUV light sourceproduces brilliant and intense VUV light. The novel VUV light source was coupled to a compact and mobile time-of-flight mass spectrometer (TOFMS). A special interface design, including optical (VUV optics) as well as electronic measures (e.g., pulsed ion extraction) was realized. The use of the excimer VUV lamp for SPI will allow the realization of very compact, rugged, and sensitive SPI-TOFMS devices, which preferably will be adapted for process analytical application or monitoring issues (e.g., chemical warfare detection). The excimer VUV-lamp technology delivers VUV light with a good beam quality and high-output power at low costs. Furthermore, it allows changing the emitted wavelength as well as the bandwidth of the excimer VUV lamp in t he 100-200-nm region by changing the gas filling. Consequently, SPI-TOFMS with an excimer light source is a fast detection technique that can be used for online monitoring, for example, in environmental studies or industrial manufacturing processes. In this paper, technology and characteristics of the new excimer light source, as well as the combination with the TOFMS, are presented. Furthermore, a first characterization of the SPI-TOFMS instrument, regarding analytical parameters such as detection limits and selectivity, is given. This includes a discussion of potential improvements that probably will be achievable within a future prototype genertation. Finally, first applications of the system for on-line measurement of organic trace species in a complex gas mixture (here, motorcycle exhaust gas) are presented.

The Neurospora rca-1 gene complements an Aspergillus flbD sporulation mutant but has no identifiable role in Neurospora sporulation.

The Aspergillus nidulans flbD gene encodes a protein with a Myb-like DNA-binding domain that is proposed to act in concert with other developmental regulators to control initiation of conidiophore development. We have identified a Neurospora crassa gene called rca-1 (regulator of conidiation in Aspergillus) based on its sequence similarity to flbD. We found that N. crassa rca-1 can complement the conidiation defect of an A. nidulans flbD mutant and that induced expression of rca-1 caused conidiation in submerged A. nidulans cultures just as was previously observed for overexpression of flbD. Thus, the N. crassa gene appears to be a functional homologue of A. nidulans flbD and this is the first demonstration of functional complementation of an A. nidulans sporulation defect using a gene from an evolutionarily distant fungus. However, deletion of the rca-1 gene in N. crassa had no major effect on growth rate, macroconidiation, microconidiation, or ascospore formation. The only phenotype displayed by the rca-1 mutant was straight or counterclockwise hyphal growth rather than the clockwise spiral growth observed for wild type. Thus, if rca-1 is involved in N. crassa development, its role is subtle or redundant.

Asexual sporulation in Aspergillus nidulans.

The formation of mitotically derived spores, called conidia, is a common reproductive mode in filamentous fungi, particularly among the large fungal class Ascomycetes. Asexual sporulation strategies are nearly as varied as fungal species; however, the formation of conidiophores, specialized multicellular reproductive structures, by the filamentous fungus Aspergillus nidulans has emerged as the leading model for understanding the mechanisms that control fungal sporulation. Initiation of A. nidulans conidiophore formation can occur either as a programmed event in the life cycle in response to intrinsic signals or to environmental stresses such as nutrient deprivation. In either case, a development-specific set of transcription factors is activated and these control the expression of each other as well as genes required for conidiophore morphogenesis. Recent progress has identified many of the earliest-acting genes needed for initiating conidiophore development and shown that there are at least two antagonistic signaling pathways that control this process. One pathway is modulated by a heterotrimeric G protein that when activated stimulates growth and represses both asexual and sexual sporulation as well as production of the toxic secondary metabolite, sterigmatocystin. The second pathway apparently requires an extracellular signal to induce sporulation-specific events and to direct the inactivation of the first pathway, removing developmental repression. A working model is presented in which the regulatory interactions between these two pathways during the fungal life cycle determine whether cells grow or develop.

Dominant mutations affecting both sporulation and sterigmatocystin biosynthesis in Aspergillus nidulans.

The initiation of conidiophore development in the filamentous fungus Aspergillus nidulans is a complex process requiring the activities of several genes including fluG, flbA, flbB, flbC, flbD, and flbE. Recessive mutations in any one of these genes result in greatly reduced expression of the brlA developmental regulatory gene and a colony morphology described as fluffy. These fluffy mutants have somewhat diverse phenotypes but generally grow as undifferentiated masses of vegetative hyphae to form large cotton-like colonies. In this paper we describe a genetic screen to identify dominant mutations resulting in similar fluffy colony morphologies. We have identified 36 dominant fluffy mutant strains and shown that 29 of these mutants have greatly reduced brlA expression as compared to wild-type. In addition, we have found that 19 of these mutants are not only developmentally altered but also fail to produce the toxic, carcinogenic, secondary metabolite sterigmatocystin. At least three of the mutants isolated result from dominant activating mutations in fadA which encodes the G alpha subunit of a heterotrimeric G-protein. Another of the mutants results from a dominant interfering mutation in brlA. We discuss the approaches taken to characterize these potentially important regulators of growth, development and secondary metabolism.

In vivo modulated N-acyl side chain of N-acetylneuraminic acid modulates the cell contact-dependent inhibition of growth.

Sialylation of plasma membrane glycoproteins is thought to be involved in the regulation of differentiation and in the process of tumorigenesis. Here we show that sialylation also affects cell-cell contact-dependent growth regulation. When cultured in the presence of non-physiological synthetic sialic acid precursors, human diploid fibroblasts no longer exhibited density-dependent inhibition of growth. Concomitantly, increased sialylation of contactinhibin, a glycoprotein involved in density-dependent inhibition of growth, was observed. These results indicate that sialidase-resistant sialic acid modifications lead to dysregulated growth control. The modifications have been induced by N-propanoyl and other N-acyl derivatives of D-mannosamine.

The Aspergillus FlbA RGS domain protein antagonizes G protein signaling to block proliferation and allow development.

flbA encodes an Aspergillus nidulans RGS (regulator of G protein signaling) domain protein that is required for control of mycelial proliferation and activation of asexual sporulation. We identified a dominant mutation in a second gene, fadA, that resulted in a very similar phenotype to flbA loss-of-function mutants. Analysis of fadA showed that it encodes the alpha-subunit of a heterotrimeric G protein, and the dominant phenotype resulted from conversion of glycine 42 to arginine (fadA(G42R)). This mutation is predicted to result in a loss of intrinsic GTPase activity leading to constitutive signaling, indicating that activation of this pathway leads to proliferation and blocks sporulation. By contrast, a fadA deletion and a fadA dominant-interfering mutation (fadA(G203R)) resulted in reduced growth without impairing sporulation. In fact, the fadA(G203R) mutant was a hyperactive asexual sporulator and produced elaborate sporulation structures, called conidiophores, under environmental conditions that blocked wild-type sporulation. Both the fadA(G203R) and the fadA deletion mutations suppressed the flbA mutant phenotype as predicted if the primary role of FlbA in sporulation is in blocking activation of FadA signaling. Because overexpression of flbA could not suppress the fadA(G42R) mutant phenotype, we propose that FlbA's role in modulating the FadA proliferation signal is dependent upon the intrinsic GTPase activity of wild-type FadA.

Mapping striate and extrastriate visual areas in human cerebral cortex.

Functional magnetic resonance imaging (fMRI) was used to identify and map the representation of the visual field in seven areas of human cerebral cortex and to identify at least two additional visually responsive regions. The cortical locations of neurons responding to stimulation along the vertical or horizontal visual field meridia were charted on three-dimensional models of the cortex and on unfolded maps of the cortical surface. These maps were used to identify the borders among areas that would be topographically homologous to areas V1, V2, V3, VP, and parts of V3A and V4 of the macaque monkey. Visually responsive areas homologous to the middle temporal/medial superior temporal area complex and unidentified parietal visual areas were also observed. The topography of the visual areas identified thus far is consistent with the organization in macaque monkeys. However, these and other findings suggest that human and simian cortical organization may begin to differ in extrastriate cortex at, or beyond, V3A and V4.

flbD encodes a Myb-like DNA-binding protein that coordinates initiation of Aspergillus nidulans conidiophore development.

The timing of asexual fruiting body formation during Aspergillus nidulans colony development is precisely regulated so that conidiophores are typically produced 1-2 mm behind the growing edge of the colony. Mutations in any of four A. nidulans genes, flbB, flbC, flbD, or flbE, result in colonies that are delayed at least 24 hr in their ability to initiate conidiophore development resulting in fluffy colonies with conidiophores forming in the center, at least 12-15 mm behind the growing edge. The requirement for each of these four genes in determining the timing of developmental initiation precedes transcriptional activation of the primary developmental regulatory gene brlA, indicating a possible role for each gene in developmentally regulated activation of brlA expression. The wild-type flbD gene was isolated and shown to encode an approximately 1.6-kb mRNA that is present throughout the A. nidulans life cycle. The deduced FlbD protein sequence predicts a 314-amino-acid polypeptide with significant identity at its amino terminus to the DNA-binding domain of the Myb family of transcription factors indicating that FlbD probably functions as a sequence-specific transcriptional activator. Although conidiophore development does not normally occur in submerged culture, forced overexpression of flbD in submerged hyphae caused inappropriate activation of brlA expression and resulted in production of complex conidiophores that produced all of the distinct cell types observed in wild-type conidiophores including viable spores. This ability of flbD overexpression to activate conidiation requires brlA, flbB, and flbA (another early developmental regulator) but does not require flbC or flbE. We propose that FlbD functions during normal development by activating transcription of other genes required for development (such as brlA) and that FlbD activity is normally controlled post-transcriptionally by an unknown mechanism.

Genetic requirements for initiating asexual development in Aspergillus nidulans.

Conidiation in the filamentous ascomycete Aspergillus nidulans requires activation of brlA, a well-characterized transcriptional regulator of genes that are induced specifically during asexual development. We have isolated and characterized developmental mutations in six loci, designated fluG, flbA, flbB, flbC, flbD, and flbE, that result in defective development and reduced brlA expression. These mutants grow indeterminately to produce masses of aerial hyphae resulting in the formation of cotton-like colonies with a "fluffy" morphology. The results of growth and epistasis tests involving all pairwise combinations of fluffy mutations indicate complex hierarchical relationships among these loci. We discuss these genetic interactions and propose that there are multiple mechanisms for activating brlA.

Progressive muscular dystrophy (Duchenne): biochemical studies by flow-cytometry.

The peanut lectin (PNL) receptor density of the cell membrane and several metabolic parameters of cultured fibroblasts of normal human individuals and of patients with muscular dystrophy were measured by simultaneous two and three parameter flow cytometry. The PNL-receptor density was significantly decreased on muscular dystrophy fibroblasts (between 20.7 and 33.6%) as compared to normal fibroblasts. The cell volume, the esterase activity, the intracellular pH, and the percentage of proliferating cells of both types of fibroblasts were not significantly altered. The mean cell volume of different fibroblast cultures varied between 2500 and 6000 micron 3. The concentration of the intracellular esterase activity of fibroblasts was low (0.169 relative units) as compared to lymphocytes and granulocytes of the peripheral blood (1.56 and 2.17 relative units). The fibroblasts had an acidic intracellular pH of 6.52 while lymphocytes and granulocytes had basic pH values of 7.30 and 7.17. Some of the fibroblasts were in the S + G2/M phase of the cell cycle (20%). The study shows that the measurement of biochemical parameters of vital and fixed single fibroblasts by flow-cytometry is of great interest for the recognition of differences between normal individuals and muscular dystrophy patients.