Competition by Bradyrhizobium Strains for Nodulation of the Nonlegume Parasponia andersonii

Bradyrhizobium strains isolated from the nonlegume Parasponia spp. formed a group of strains that were highly competitive for nodulation of P. andersonii when paired with strains isolated from legumes. Strains from legumes, including those of similar effectiveness to NGR231 and CP283, were not able to form nodules as single occupants on P. andersonii in the presence of Parasponia strains. However, NGR86, an isolate from Macroptilium lathyroides, jointly occupied one-third of the nodules formed with each of the three strains isolated from Parasponia spp. Time taken for nodules to appear may have influenced the outcome of competition, since CP283 and all isolates from legumes were slow to nodulate P. andersonii. Among the Parasponia strains, competitiveness for nodulation of P. andersonii was not associated with effectiveness of nitrogen fixation. The highly effective strain CP299 was a poor competitor when paired with the least effective strain NGR231. CP283 was the least competitive of the Parasponia strains but was still able to dominate nodules when paired with legume isolates. Dual occupancy was high, up to 67% when the inoculum contained CP299 and CP273. Both the Muc⁺ and Muc^- types of CP283 form a symbiosis of similar effectiveness and were similarly competitive at high inoculation densities, but the Muc^- form was more competitive at low inoculum densities. Both forms frequently occupied the same nodule. Bradyrhizobium strains isolated from Parasponia spp. may have specific genetic information that favor their ability to competitively and effectively infect plants in the genus Parasponia (Ulmaceae) outside the Leguminosae.     

A Novel GFP-Fused Eukaryotic Membrane Protein Expression System in Lactococcus lactis and Its Application to Overexpression of an Elongase

Ultraviolet (UV) irradiation has high potential to inactivate a wide range of biologic agents and is one of several nonadditive technologies being studied. The photoinactivation property of pulsed UV laser radiation (at wavelengths of 355 and 266 nm), used as an effective physical means to inactivate two typical microorganisms, prokaryotic (Escherichia coli K12) and eukaryotic (Saccharomyces cerevisiae), with respect to dose and exposure times, was examined. An E. coli population of 1.6 × 10⁴ colony-forming units (CFU)/ml was inactivated with a dose of 16.7 J/cm² energy at 355-nm wavelength. However, E. coli cells at higher concentrations were inactivated by only 98% using the same dose. Interestingly, an E. coli population of 2 × 10⁷ CFU/ml was completely inactivated using only 0.42 J/cm² at 266-nm wavelength (P ≤ 0.05). With respect to S. cerevisiae, the results were similar to those of E. coli irradiation considering that S. cerevisiae is 100 times larger than E. coli. A dose of 16.7 J/cm² completely inactivated an S. cerevisiae population of 6 × 10³ CFU/ml at 355-nm wavelength. Exposure to 266-nm wavelength, with energy doses of 1.67, 0.835, and 0.167 J/cm², successfully inactivated S. cerevisiae populations of 3 × 10⁶, 1.4 × 10⁵, and 1.5 × 10⁴ CFU/ml, respectively (P ≤ 0.05). In conclusion, compared with 355-nm wavelength, a pulsed UV laser at 266-nm wavelength inactivated a high titer of bacterial and yeast indicator standards suspended in phosphate-buffered saline-A.     

Variation of chromophoric dissolved organic matter and possible attenuation depth of ultraviolet radiation in Yunnan Plateau lakes

The increase of ultraviolet radiation (UVR, 280–400 nm) caused by stratospheric ozone depletion has profound effects on aquatic ecosystems. High-altitude lakes in the Yunnan Plateau are exposed to high intensities of UVR and contain low concentrations of chromophoric dissolved organic matter (CDOM). Thirty-eight lakes in the Yunnan Plateau with elevations from 1291 to 3809 m above sea level were investigated to study CDOM concentrations and possible effects of UVR on the lake ecosystem. The attenuation of UVR in the Yunnan Plateau lakes was calculated from the absorption coefficient of CDOM based on an empirical relationship from lakes in the Alps and Pyrenees mountains. Absorption coefficients [α(λ)] at 320 nm [α(320)] ranged from 0.52 to 14.05 m⁻¹ (mean ± standard deviation, 4.40 ± 3.85 m⁻¹) and at 380 nm [α(380)] from 0.05 to 4.51 m⁻¹ (1.40 ± 1.30 m⁻¹). The exponential slope coefficient for the relationship of wavelength to α(λ) ranged from 16.2 to 41.4 μm⁻¹ (21.74 ± 4.93 μm⁻¹) over the 280–400 nm interval. Normalized fluorescence emission (NFLU) at 450 nm from an excitation wavelength of 355 nm, F _n(355), averaged 7.93 ± 3.22 NFLU. A significant positive relationship was found between α(355) and F _n(355). The estimated diffuse attenuation coefficients of UV-B (320 nm) and UV-A (380 nm) ranged from 0.55 to 15.77 m⁻¹ and from 0.24 to 6.73 m⁻¹; the corresponding 1% attenuation depths ranged from 0.29 to 8.44 m and from 0.68 to 19.12 m. Twenty-five of 38 lakes had 1% UV-B attenuation depths of 1.5 m or more. The median 1% attenuation depth was 28.8% of the sampling depth for UV-B radiation and 60% for UV-A. In addition to CDOM, chlorophyll α (Chla) and total suspended matter (TSM) also may contribute to attenuation of UVR.     

Spectral Requirements for Flower Initiation in Two Longday Plants, Rape (Brassica campestris cv. Ceres), and Spring Wheat (Triticum×aestivum)

Action spectra for the promotion of flowering by long periods of irradiation in the red and far-red regions of the spectrum have been determined by the use of interference filters. The percentage floral initiation was greatest at 710–720 um for both wheat and rape (about 1 k erg · cm^?2· s^?1). Lowering the temperature from 35 to 15°C did not shift the most effective wavelength, suggesting that the effect of high levels of radiation at 710 nm was not related to a balance between photoactivation of phytochrome and its destruction. In both wheat and rape, an increase in temperature promoted the flower-initiating action of broad-band far-red and 710 nm radiation more than the action of broadband red or 660 nm radiation. The flower-promoting effect of broad-band red and 600 nm radiation was particularly depressed by temperatures above 25°C. It is concluded that the promotion of flowering by long periods of high energy at 710 nm is caused by the activity of a high-energy pigment, rather than through a special effect of phytochrome.     

Ultraviolet disinfection of Schistosoma mansoni cercariae in water

BackgroundSchistosomiasis is a parasitic disease that is transmitted by skin contact with waterborne schistosome cercariae. Mass drug administration with praziquantel is an effective control method, but it cannot prevent reinfection if contact with cercariae infested water continues. Providing safe water for contact activities such as laundry and bathing can help to reduce transmission. In this study we examine the direct effect of UV light on Schistosoma mansoni cercariae using ultraviolet light-emitting diodes (UV LEDs) and a low-pressure (LP) mercury arc discharge lamp.MethodologyS. mansoni cercariae were exposed to UV light at four peak wavelengths: 255 nm, 265 nm, 285 nm (UV LEDs), and 253.7 nm (LP lamp) using bench scale collimated beam apparatus. The UV fluence ranged from 0–300 mJ/cm² at each wavelength. Cercariae were studied under a stereo-microscope at 0, 60, and 180 minutes post-exposure and the viability of cercariae was determined by assessing their motility and morphology.ConclusionVery high UV fluences were required to kill S. mansoni cercariae, when compared to most other waterborne pathogens. At 265 nm a fluence of 247 mJ/cm² (95% confidence interval (CI): 234–261 mJ/cm²) was required to achieve a 1-log₁₀ reduction at 0 minutes post-exposure. Cercariae were visibly damaged at lower fluences, and the log reduction increased with time post-exposure at all wavelengths. Fluences of 127 mJ/cm² (95% CI: 111–146 mJ/cm²) and 99 mJ/cm² (95% CI: 85–113 mJ/cm²) were required to achieve a 1-log₁₀ reduction at 60 and 180 minutes post-exposure at 265 nm. At 0 minutes post-exposure 285 nm was slightly less effective, but there was no statistical difference between 265 nm and 285 nm after 60 minutes. The least effective wavelengths were 255 nm and 253.7 nm. Due to the high fluences required, UV disinfection is unlikely to be an energy- or cost-efficient water treatment method against schistosome cercariae when compared to other methods such as chlorination, unless it can be demonstrated that UV-damaged cercariae are non-infective using alternative assay methods or there are improvements in UV LED technology.     

D-pantolactone as a circular dichromism (CD) calibration.

A critical study of circular dichromism (CD) measurement at the wavelength region around 220 nm was made. Test sample methods gave deviations of abou 20%, at most, between instruments calibrated at 290 nm with D-10-camphorsulfonic acid. A calculation of the CD from optical rotatory dispersion (ORD) data was made for d-pantolactone which gave a CD maximum at 219 nm, [θ]₂₁₉²⁵ = ?17.3 × 10³ in water. A proposal is made to use this value for calibration of CD in the 220-nm region.     

Using UVC Light-Emitting Diodes at Wavelengths of 266 to 279 Nanometers To Inactivate Foodborne Pathogens and Pasteurize Sliced Cheese

UVC light is a widely used sterilization technology. However, UV lamps have several limitations, including low activity at refrigeration temperatures, a long warm-up time, and risk of mercury exposure. UV-type lamps only emit light at 254 nm, so as an alternative, UV light-emitting diodes (UV-LEDs) which can produce the desired wavelengths have been developed. In this study, we validated the inactivation efficacy of UV-LEDs by wavelength and compared the results to those of conventional UV lamps. Selective media inoculated with Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes were irradiated using UV-LEDs at 266, 270, 275, and 279 nm in the UVC spectrum at 0.1, 0.2, 0.5, and 0.7 mJ/cm², respectively. The radiation intensity of the UV-LEDs was about 4 μW/cm², and UV lamps were covered with polypropylene films to adjust the light intensity similar to those of UV-LEDs. In addition, we applied UV-LED to sliced cheese at doses of 1, 2, and 3 mJ/cm². Our results showed that inactivation rates after UV-LED treatment were significantly different (P < 0.05) from those of UV lamps at a similar intensity. On microbiological media, UV-LED treatments at 266 and 270 nm showed significantly different (P < 0.05) inactivation effects than other wavelength modules. For sliced cheeses, 4- to 5-log reductions occurred after treatment at 3 mJ/cm² for all three pathogens, with negligible generation of injured cells.     

Strong Near-infrared Avalanche Photoluminescence from Ag Nanowire Arrays

Ag nanowire (NW) arrays with NW diameter d _NW?=?12–120 nm were electrodeposited in anodic aluminum oxide templates. Strong avalanche photoluminescence (PL) from Ag NW arrays with small d _NW were observed near 914 nm by using picosecond laser at the excitation wavelength 808 nm, which is originated from the plasmon-enhanced radiative intraband transitions. The peak PL intensity of the avalanche PL from the sample with small diameter d _NW?=?12 nm is about 10² times stronger than that of the linear PL from the sample with large diameter d _NW?=?120 nm. The opposite excitation polarization dependence and emission polarization distribution of the PL from Ag NW array with d _NW?=?12 nm and d _NW?=?120 nm were also observed.     

In Vitro Comparison of Hypericin and 5-Aminolevulinic Acid-Derived Protoporphyrin IX for Photodynamic Inactivation of Medulloblastoma Cells

Background

Hypericin (HYP) is a naturally occurring photosensitizer. Cellular uptake and photodynamic inactivation after incubation with this photosensitizer have neither been examined in medulloblastoma cells in vitro, nor compared with 5-aminolevulinic acid-derived protoporphyrin IX (5-ALA-derived PpIX).

Methods

In 3 medulloblastoma cell lines (D283 Med, Daoy, and D341 Med) the time- and concentration-dependent intracellular accumulation of HYP and 5-ALA-derived PpIX was analyzed by fluorescence microscopy (FM) and FACS. Photocytotoxicity was measured after illumination at 595 nm (HYP) and 635 nm (5-ALA-derived PpIX) in D283 Med cells and compared to U373 MG glioma cells.

Results

All medulloblastoma cell lines exhibited concentration- and time-dependent uptake of HYP. Incubation with HYP up to 10 µM resulted in a rapid increase in fluorescence intensity, which peaked between 2 and 4 hours. 5-ALA-derived PpIX accumulation increased in D283 Med cells by 22% over baseline after 5-ALA incubation up to 1.2 mM. Photocytotoxicity of 5-ALA-derived PpIX was higher in D283 Med medulloblastoma compared to U373MG glioma. The [lethal dose (light dose that is required to reduce cell survival to 50% of control)] of 5-ALA-derived PpIX was 3.8 J/cm² in D283 Med cells versus 5.7 J/cm2 in U373MG glioma cells. Photocytotoxicity of HYP in D283 Med cells was determined at 2.5 µM after an incubation time of 2 h and an illumination wavelength of 595 nm. The value was 0.47 J/cm².

Conclusion

By its 5-fold increase in fluorescence over autofluorescence levels HYP has excellent properties for tumor visualization in medulloblastomas. The high photocytotoxicity of HYP, compared to 5-ALA-derived PpIX, is convincingly demonstrated by its 8- to 13-fold lower . Therefore HYP might be a promising molecule for intraoperative visualization and photodynamic treatment of medulloblastomas.     

Sugar‐feeding status alters biting midge photoattraction

The biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae) transmits pathogens to both livestock and wildlife. Biting midge surveillance relies heavily on light traps for collection; however, little is known about the light spectra preferences of C. sonorensis midges. A light assay arena was constructed and light‐emitting diodes (LEDs) of various light spectra were used as light sources to evaluate midge photoattraction. A comparison of responses to light spectra indicated the highest proportions of C. sonorensis were attracted to ultraviolet (UV) light and that midges differentiated 10‐nm differences in wavelength. Stronger intensities of UV light resulted in greater attraction. Midges exhibited both sugar‐seeking and escape behaviours under different conditions of sugar supplementation before and during the experiment. These behaviours occurred with lights of 355 nm and 365 nm in wavelength. Based on the results of this study, the attraction of C. sonorensis to light traps can be improved through the use of bright LEDs at 355 nm or 365 nm.     

Electrophysiology of the visual system in the cricket Gryllus firmus (Orthoptera: Gryllidae): Spectral sensitivity of the compound eyes

The electroretinographic spectral sensitivity of the cricket compound eyes shows the presence of two receptor types, a dominant one at 520 nm and another in the near-u.v. (λ_max 355 ± 5 nm) under dark- and intense chromatic adaptation conditions (Fig. 3). The waveform of the electrical responses elicited by short-wavelength stimuli differ from those elicited by long wavelength stimuli (Fig. 1).     

Analysis of photoernzymatic repair of UV lesions in DNA by single light flashes. XI. Light-induced activation of the yeast photoreactivating enzyme

Photoreactivating enzyme (PRE) from yeast (as semi-crude extract, or in highly purified form) shows increased activity if its is illuminated with near UV or short wavelength visible light prior to its use for photoenzymatic repair of UV-induced pyrimidine dimers in transforming DNA in vitro. This effect results from an alternation in PRE molecules changing those with low activity in the light-dependent step of the reaction to a higher activity. Light-induced activation of PRE preparations is slowly lost by dark storage for several hours to 1 day (faster at 23°C than at 5°C), but can be recovered repeatedly by renewed preillumination. The action spectrum for these preillumination effects generally resembles that for the photoenzymatic repair reaction itself, having its maximum in the same 355–385 nm region as the latter, but light of somewhat longer wavelengths (546 nm) is still effective. Preilluminated PRE is also more stable to thermal inactivation (65°C) than untreated enzyme.     

Action Spectra for Conversions of Phycochrome c from Nostoc muscorum

The reversibly photochromic pigment, phycochrome c, was extracted from the blue-green alga Nostoc muscorum strain A. Action spectra were determined for in vitro conversions of the pigment from the short wavelength to the long wavelength form and vice versa. The action peak for the absorbance decrease at 650 nm is at 630 nm. During this decrease there is only a slight increase of the absorbance in the green region. Green and yellow light (maximum efficiency at 580 nm) completely restores absorbance at 650 nm. The observations are explained by the existence of three spectrally different forms of phycochrome c: P^c₆₃₀ and P^c₆₅₀ which equilibrate in darkness and P^c₅₈₀ which is reversibly photoconvertible to P^c₆₃₀. We have also measured the absorbance changes brought about by saturating irradiations with light of various wavelengths (“photostationary state spectrum”). Extreme photostationary states were obtained with about 650 nm and 500 nm light.     

Spectral Sensitivity of Melanophores in the Primary Color Response of the Rose Bitterling, <Emphasis Type="Italic">Rhodeus ocellatus ocellatus</Emphasis>

Melanophores of young Rhodeus ocellatus ocellatus have the ability to respond by melanosome dispersion to the direct action of visible light. The effective wavelength within visible light region for inducing melanosome dispersion was investigated using melanophores located around the base of the dorsal fin of young fish set on the stage of a light microscope. The melano- -’ phores were exposed to light of various wavelengths (420–680 nm) but of the same intensity by placing interference filters under the condenser diaphragm. The most effective wavelength was about 420 nm. Longer wavelengths were less effective for the induction of melanosome dispersion.     

Spectral Characteristics of Near-Infrared Surface Plasmon Resonance

Intrinsic properties of surface plasmons (SPs) excited with Kretschmann configuration were analyzed as a function of wavelength, including the propagation length, the penetration depth, the Goos–Hänchen (GH) shift, and the field enhancement. The calculated results indicate that there exists a critical thickness (t _cr) of the gold layer and that the maximum GH shift occurring exactly at the SP resonance wavelength (λ _R) rapidly varies from positive to negative with changing of the gold layer thickness from t?<?t _cr to t?>?t _cr. The maximum field enhancement happens not at λ _R but at a wavelength smaller than λ _R due to the phase retardation between the transmitted and reflected light. Simulations also reveal that a broadband collimated near-infrared beam can simultaneously excite two SPs with different responses to a refractive index (RI) change: the shorter-wavelength SP able to make a small redshift and the longer-wavelength SP capable of yielding a large blueshift. Only the shorter-wavelength SP was experimentally observed and its RI sensitivity was measured to increase from 3,539 nm/RIU at λ _R?=?707.6 nm to 57,143 nm/RIU at λ _R?=?1,398 nm. The SP at λ _R?=?1,013 nm moved to λ _R?=?1,029 nm in response to the saturation adsorption of bovine serum albumin, and the corresponding surface coverage was determined to be Γ?=?1.565 ng/mm² based on a quasilinear dependence of Γ on the resonance wavelength shift (?λ _R) deduced theoretically. Butyrylcholinesterase adsorption from a dilute solution of 10 nM protein in phosphate buffer solution leads to a redshift of ?λ _R?=?10 nm, corresponding to Γ?≈?0.97 ng/mm².     

Flavodoxin from the blue-green alga Nostoc strain MAC

A flavodoxin was isolated from the blue-green alga Nostoc strain MAC grown photoautotrophically or chemoheterotrophically in iron-deficient medium. In vitro, the flavodoxin would support NADP⁺ photoreduction by photosynthetic membranes, pyruvate oxidation by the phosphoroclastic system of Clostridium pasteurianum, and electron transfer to Cl. pasteurianum hydrogenase. In its oxidized form, the flavodoxin had absorbance maxima at 274 sh283 sh293, 376 sh432 and 466 sh488 nm. Reduction by dithionite proceeded via a neutral, blue semiquinone radical. The flavodoxin contained 1 mol of FMN per mol of protein and the amino acid composition showed a predominance of acidic residues; cysteine was apparently absent. A minimum MW of ca 22 000 derived from these data was confirmed by electrophoresis on SDS-polyacrylamide gels and by ultracentrifugal analysis. This flavodoxin thus belongs to the higher MW group of these low potential electron transfer proteins.     

Spectral Dependence of Photoregulation of Inorganic Nitrogen Metabolism in Chlamydomonas reinhardii

The utilization of NO₃⁻ by green algae growing photoautotrophically under air, which are growth conditions close to their more habitual situations in nature, is associated with the excretion of NO₂⁻ and NH₄⁺ to the culture medium. The entire process is promoted by blue light and depends on photosynthetically active radiation for the required reducing equivalents. The stimulation of NO₃⁻ utilization and of its associated NO₂⁻ and NH₄⁺ excretions saturated at very low quantum fluxes of blue light (15 microequivalents per square meter per second) in Chlamydomonas reinhardii cells sparged with CO₂-free air and irradiated with 50 microequivalents per square meter per second background red light. The wavelength dependence data of this stimulation correlated closely with the in situ photoactivation of nitrate reductase and also with the light induced increase in its biosynthesis and/or assembly.

These results indicate that the photoregulation of inorganic N metabolism in C. reinhardii is mainly due to the blue light modulation of nitrate reductase. Although flavins are the most suitable candidates to act as physiological photoreceptors, the wavelength dependence data only show a major peak in the blue region between 400 and 500 nanometers.

     

Water Transport across Maize Roots : Simultaneous Measurement of Flows at the Cell and Root Level by Double Pressure Probe Technique

A double pressure probe technique was used to measure simultaneously water flows and hydraulic parameters of individual cells and of excised roots of young seedlings of maize (Zea mays L.) in osmotic experiments. By following initial flows of water at the cell and root level and by estimating the profiles of driving forces (water potentials) across the root, the hydraulic conductivity of individual cell layers was evaluated. Since the hydraulic conductivity of the cell-to-cell path was determined separately, the hydraulic conductivity of the cell wall material could be evaluated as well (Lp_cw = 0.3 to 6.10⁻⁹ per meter per second per megapascal). Although, for radial water flow across the cortex and rhizodermis, the apoplasmic path was predominant, the contribution of the hydraulic conductance of the cell-to-cell path to the overall conductance increased significantly from the first layer of the cortex toward the inner layers from 2% to 23%. This change was mainly due to an increase of the hydraulic conductivity of the cell membranes which was Lp = 1.9.10⁻⁷ per meter per second per megapascal in the first layer and Lp = 14 to 9.10⁻⁷ per meter per second per megapascal in the inner layers of the cortex. The hydraulic conductivity of entire roots depended on whether hydrostatic or osmotic forces were used to induce water flows. Hydrostatic Lp_r was 1.2 to 2.3.10⁻⁷ per meter per second per megapascal and osmotic Lp_r = 1.6 to 2.8.10⁻⁸ per meter per second per megapascal. The apparent reflection coefficients of root cells (σ_s) of nonpermeating solutes (KCI, PEG 6000) decreased from values close to unity in the rhizodermis to about 0.7 to 0.8 in the cortex. In all cases, however, σ_s was significantly larger than the reflection coefficient of entire roots (σ_sr). For KCI and PEG 6000, σ_sr was 0.53 and 0.64, respectively. The results are discussed in terms of a composite membrane model of the root.     

Luminescence of bacteriorhodopsin from Halobacterium halobium and its connection with the photochemical conversions of the chromophore

Red luminescence of purple membranes from Halobacterium halobium cells in suspension, dry film or freeze-dried preparations was studied and its emission, excitation and polarization spectra are reported. The emission spectra have three bands at 665–670, 720–730 and at 780–790 nm. The position (maximum at 580 nm) and shape of the excitation spectra are close to those of the absorption spectra. The spectra depend on experimental conditions, in particular on pH of the medium. Acidification increases the long wavelength part of the emission spectra and shifts the main excitation maximum 50–60 nm to the longer wavelength side. Low-temperature light-induced changes of the absorption, emission and excitation spectra are presented. Several absorbing and emitting species of bacteriorhodopsin are responsible for the observed spectral changes. The bacteriorhodopsin photoconversion rate constant was estimated to be about 1 · 10¹¹ s^?1 at ? 196°C from the quantum yields of the luminescence (1 · 10^?3) and photoreaction (1 · 10^?1). The temperature dependence of the luminescence quantum yield points to the existence of two or three quenching processes with different activation energies. High degree of luminescence polarization (about 45–47%) throughout the absorption and fluorescence spectra and its temperature independence show that there is no energy transfer between bacteriorhodopsin molecules and no chromophore rotation during the excitation lifetime. In carotenoid-containing membranes, energy migration from the bulk of carotenoids to bacteriorhodopsin was not found either. Bacteriorhodopsin phosphorescence was not observed in the 500–1100 nm region and the emission is believed to be fluorescence by nature.