Upper boundary of the biosphere.

By using meterological rockets fitted with specially designed analyzers, samples for microbiological investigation have been taken. The analyzer design prevented extraneous microorganisms from penetrating into the analyzer. Before being used, the analyzers were sterilized with high gamma-ray doses. For the first time microorganisms have been detected in the mesosphere at an altitude of 48 to 77 km. The microorganisms are microscopic fungi having black conidia or spores (Circinella muscae, Aspergillus niger, Papulaspora anomala) and one species forming green conidia (Penicillium notatum). Colonies of Mycobacterium luteum and Micrococcus albus have also grown. Five of the six species have synthesized pigments. The presence of pigmented microbial forms leads us to believe that natural selection is occurring in the mesosphere because cells possessing chromogenous pigments (carotenoids, melanins) are more resistant to ultraviolet-ray action. A greater number of microorganisms have been registered in the mesosphere during dust storms than in the absence of strong winds.     

Isolation and characterization of marine pigmented bacteria from Norwegian coastal waters and screening for carotenoids with UVA-blue light absorbing properties

Microbial culture collections are important resources for isolation of natural compounds with novel properties. In this study, a culture collection of around 1,500 pigmented heterotrophic bacteria was established. The bacteria were isolated from the sea surface microlayer at different sampling sites along the mid-part of the Norwegian coast. The bacterial isolates produced pigments of various coloration (e.g. golden, yellow, red, pink and orange). Methanol extracts of sixteen isolates were characterized with LC-Diodearray-TOF mass spectrometry analysis. The number of pigments per isolate varied considerably, and a tentative identification of the pigments was performed based on UV-absorbance profile and molecular formula assignation based on the accurate mass determination. The LC-MS analyses revealed that most of the pigments probably were carotenoids. Furthermore, we developed a high throughput LC-MS method for characterization and screening of a larger sub-fraction (300 isolates) of the culture collection. The aim was to screen and identify bacterial isolates producing carotenoids that absorb light in the UVA-Blue light. Six of the bacterial strains were selected for detailed investigation, including 16s rRNA sequencing, preparative HPLC for purification of major carotenoids and subsequent structural elucidation with NMR. Among the identified carotenoids were zeaxanthin, nostoxanthin and sarcinaxanthin, some with novel glycosylation patterns.     

Sample-hold technique for analysis of respiratory gas composition at high breathing frequencies

A gas sampling device is described for continuous monitoring of respiratory gas composition that is applicable to experimental conditions when the breathing frequency is very high (greater than 2 Hz) and the response time of conventional gas analyzers becomes a critical limiting factor. The system utilizes the principle of discontinuous gas collection at any selected point of the respiratory cycle facilitated by ultraspeed piezoelectric valves and includes provision for sample-hold characteristics. Two distinct modes of operation are supported. In phase-locked mode gas sampling is synchronous with breathing frequency. In scanning mode gas collection is asynchronous with breathing frequency. Phase-locked mode may be used for continuous monitoring of end-tidal gas concentrations, whereas scanning mode is intended for assessing the gas concentration profile throughout the respiratory cycle. The system may be applied to steady breathing encountered in mechanical ventilation at high frequency or during quasi-steady breathing observed in panting animals. Combined with a respiratory mass spectrometer, the system has been used for measurement of gas concentrations in alveolar gas mixtures at breathing frequencies ranging from 3 to 30 Hz that were otherwise not amenable to rapid measuring techniques.     

Genetic control of chloroplast pigment development in soybeans as a function of leaf and plant maturity

Changes in the major chloroplast pigments of pigment-deficient genotypes of soybeans were studied as a function of leaf age and plant age. The trends of pigment development are plotted as a function of relative leaf age at two periods of plant development. Comparisons are made between aging leaves from different nodes and leaves of different ages from the same node. In addition, trends of pigment development are expressed as a ratio of pigment-deficient/normal genotypes for five different genotypes and seven sampling periods. Those genotypes that exhibit a lag in production of pigments during leaf development show a similar lag in overall plant pigment development.     

Regulation of the xanthophyll cycle pool size in duckweed (Lemna minor) plants

Duckweed ( Lemna minor L.) plants grown under high light are characterized, when compared to low light acclimated plants, by a higher xanthophyll cycle (VAZ) pool content, but also by a higher proportion of photoconvertible violaxanthin and a superior ability to synthesize VAZ pigments. When duckweed plants were transferred to a high light environment a general response was the quick adjustment of the carotenoid composition, mainly xanthophyll cycle pigments. These changes resulted from a balance between a process of continuous light-independent carotenoid degradation and a light-induced accumulation. The use of norflurazon, an inhibitor of carotenogenesis, allowed us to demonstrate that the observed light induced increase of the VAZ pool was mainly caused by de novo synthesis through carotenogenesis. The extent of light-induced carotenogenesis was proportional to the light treatment and also to the operation of the VAZ cycle since it was partly abolished by treatments leading to a low activity of the VAZ cycle, such as low light, DTT or DCMU. These results suggest that not only the light itself, but also a mechanism triggered by a factor associated with the de-epoxidation state of the VAZ cycle controls carotenogenesis at some point before phytoene formation in the terpenoid biosynthesis pathway.     

LH500血细胞分析仪故障检修

对LH500血细胞分析仪常见故障,如气源、进样、计数、HGB与分类等,从原理与工作流程方面进行阐述。同时分析了故障原因与排除方法,总结检修思路与经验,供同行分享。     

Irradiance prior to and during desiccation improves the tolerance to excess irradiance in the desiccated state of the old forest lichen Lobaria pulmonaria

Hydrated thalli of the lichen Lobaria pulmonaria were either preconditioned to dim irradiance (DI, 5 μmol m⁻² s⁻¹) or medium irradiance (MI, 200 μmol m⁻² s⁻¹) for 6 h. After this 6 h period, the thalli were allowed to desiccate under the two respective irradiances. Thereafter, these dry lichens were exposed to high irradiance (HI, 1 000 μmol m⁻² s⁻¹) for 60 h. After this HI treatment, the maximal photochemical quantum yield (F_V/F_M) and the de-epoxidation state of xanthophyll cycle pigments (DEPS) were highest in thalli preconditioned to MI. Hence irradiance in the last hydrated period before sampling is significant for the physiological state of lichens. A standardized irradiance pre-treatment before start of experiments is recommended.     

Steady-state polarized light spectroscopy of isolated Photosystem I complexes

Monomeric and trimeric Photosystem I core complexes from the cyanobacterium Synechocystis PCC 6803 and LHC-I containing Photosystem I (PS I-200) complexes from spinach have been characterized by steady-state, polarized light spectroscopy at 77 K. The absorption spectra of the monomeric and trimeric core complexes from Synechocystis were remarkably similar, except for the amplitude of a spectral component at long wavelength, which was about twice as large in the trimeric complexes. This spectral component did not contribute significantly to the CD-spectrum. The (77 K) steady-state emission spectra showed prominent peaks at 724 nm (for the Synechocystis core complexes) and at 735 nm (for PS I-200). A comparison of the excitation spectra of the main emission band and the absorption spectra suggested that a significant part of the excitations do not pass the red pigments before being trapped by P-700. Polarized fluorescence excitation spectra of the monomeric and trimeric core complexes revealed a remarkably high anisotropy (0.3) above 705 nm. This suggested one or more of the following possibilities: 1) there is one red-most pigment to which all excitations are directed, 2) there are more red-most pigments but with (almost) parallel orientations, 3) there are more red-most pigments, but they are not connected by energy transfer. The high anisotropy above 705 nm of the trimeric complexes indicated that the long-wavelength pigments on different monomers are not connected by energy transfer. In contrary to the Synechocystis core complexes, the anisotropy spectrum of the LHC I containing complexes from spinach was not constant in the region of the long-wavelength pigments, and decreased significantly below 720 nm, the wavelength where the long-wavelength pigments on the core complexes start to absorb. These results suggested that in spinach the long-wavelength pigments on core and LHC-I are connected by energy transfer and have a non-parallel average Q_y(0-0) transitions.Abbreviations PS Photosystem - P Primary donor - Chl chlorophyll - LHC light-harvesting complex - CD circular dichroism - LD linear dichroism - BisTris 2-[bis(2-hydroxyethyl)amino]-2-hydroxy-methylpropane-1,3-diol - RC reaction center     

Cell culture monitoring via an auto‐sampler and an integrated multi‐functional off‐line analyzer

Mammalian cell‐based bioprocesses are used extensively for production of therapeutic proteins. Off‐line monitoring of such cultivations via manual sampling is often labor‐intensive and can introduce operator‐dependent error into the process. An integrated multi‐functional off‐line analyzer, the BioProfile FLEX (NOVA Biomedical, Waltham MA) has been developed, which combines the functionality of three off‐line analyzers (a cell counter, an osmometer, and a gas/electrolyte & nutrient/metabolite bio‐profile analyzer) into one device. In addition, a novel automated sampling system has also been developed that allows the BioProfile FLEX to automatically analyze the culture conditions in as many as ten bioreactors. This is the first report on the development and function of this integrated analyzer and an auto‐sampler prototype for monitoring of mammalian cell cultures. Evaluation of the BioProfile FLEX was conducted in two separate laboratories and involved two BioProfile FLEX analyzers and two sets of reference analyzers (Nova BioProfile 400, Beckman‐Coulter Vi‐Cell AS, and Advanced Instruments Osmometer 3900), 13 CHO cell lines and over 20 operators. In general, BioProfile FLEX measurements were equivalent to those obtained using reference analyzers, and the auto‐sampler did not alter the samples it provided to the BioProfile FLEX. These results suggest that the system has the potential to dramatically reduce the manual labor involved in monitoring mammalian cell bioprocesses without altering the quality of the data obtained, and integration with a bioreactor control system will allow feedback control of parameters previously available only for off‐line monitoring. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

History of anthropogenically mediated eutrophication of Lake Peipsi as revealed by the stratigraphy of fossil pigments and molecular size fractions of pore-water dissolved organic matter

We investigated stratigraphic changes in fossil pigments and the molecular structure of the UV-absorbing fraction of pore-water dissolved organic matter in a sedimentary record from Lake Peipsi (Estonia/Russia) temporally covering the 20th century. The aims of the study were to define the onset of eutrophication in the lake and to track its course. An attempt was also made to reconstruct lake conditions before the intensive nutrient loading began. Fossil pigment analysis indicated that the eutrophication of the lake started in the 1960s and accelerated in the 1970s. Sedimentary pigments also indicate a continuing tendency of the lake ecosystem towards eutrophy in the 1980s and 1990s. However, changes in the molecular size structure of pore-water dissolved organic matter indicated that the contribution of autochthonous matter to the organic pool of the lake ecosystem had already started to increase around the end of the 1930s. We conclude that this rise was generated by a coincidence of several anthropogenic and natural factors. The pore-water data also show that a slight relative reduction in the autochthonous organic matter took place in the 1990s. A discordance in the paleodata obtained for the beginning of the 20th century complicates clear conclusions about earlier conditions in the lake. On the one hand, the qualitative characteristics of pore-water dissolved organic matter and the low concentration of chlorophyll a indicate that the phytoplankton biomass was low in Lake Peipsi during that period. On the other hand, the concentrations of marker pigments of specific phytoplankton groups are high, comparable with the values in the recent sediments. Possible reasons for the high levels of these pigments in the early 1900s sediments, such as a shift in the preservation conditions of organic substances and their transport from the lake’s catchment, are discussed.     

Dynamics of cover, UV-protective pigments, and quantum yield in biological soil crust communities of an undisturbed Mojave Desert shrubland

Biological soil crusts are an integral part of dryland ecosystems. We monitored the cover of lichens and mosses, cyanobacterial biomass, concentrations of UV-protective pigments in both free-living and lichenized cyanobacteria, and quantum yield in the soil lichen species Collema in an undisturbed Mojave Desert shrubland. During our sampling time, the site received historically high and low levels of precipitation, whereas temperatures were close to normal. Lichen cover, dominated by Collema tenax and C. coccophorum, and moss cover, dominated by Syntrichia caninervis, responded to both increases and decreases in precipitation. This finding for Collema spp. at a hot Mojave Desert site is in contrast to a similar study conducted at a cool desert site on the Colorado Plateau in SE Utah, USA, where Collema spp. cover dropped in response to elevated temperatures, but did not respond to changes in rainfall. The concentrations of UV-protective pigments in free-living cyanobacteria at the Mojave Desert site were also strongly and positively related to rainfall received between sampling times (R² values ranged from 0.78 to 0.99). However, pigment levels in the lichenized cyanobacteria showed little correlation with rainfall. Quantum yield in Collema spp. was closely correlated with rainfall. Climate models in this region predict a 3.5–4.0 °C rise in temperature and a 15–20% decline in winter precipitation by 2099. Based on our data, this rise in temperature is unlikely to have a strong effect on the dominant species of the soil crusts. However, the predicted drop in precipitation will likely lead to a decrease in soil lichen and moss cover, and high stress or mortality in soil cyanobacteria as levels of UV-protective pigments decline. In addition, surface-disturbing activities (e.g., recreation, military activities, fire) are rapidly increasing in the Mojave Desert, and these disturbances quickly remove soil lichens and mosses. These stresses combined are likely to lead to shifts in species composition and the local extirpation of some lichen or moss species. As these organisms are critical components of nutrient cycling, soil fertility, and soil stability, such changes are likely to reverberate throughout these ecosystems.     

Kinetic,isotherm and thermodynamic investigations on adsorption of trace elements and pigments from soybean oil using high voltage electric field-assisted bleaching: A comparative study

In this study, the amounts of trace elements (Fe (II) and Cu (II)) and pigments (chlorophyll and carotenoid) in soybean oil were evaluated under high voltage electric field (HVEF) bleaching method at different voltage (10 and 20 kV), temperature (35–65 °C), time (0−30 min) and clay percentage (0.5–2 %) and then were compared to the industrial bleaching method (IBM). The kinetic data of ions and pigments adsorbed on activated bentonite clay under IBM and HVEF at two voltages of 10 and 20 kV followed the mechanism of the pseudo-first-order model (PFOM). The carotenoid and chlorophyll equilibrium data followed a Freundlich isotherm type model, which demonstrated multilayer adsorption under HVEF. The thermodynamic parameters (ΔG°, ΔH°, and ΔS°) displayed that the adsorption of trace metal ions and pigments on bentonite clay under IBM and HVEF were feasible, endothermic and spontaneous between 35 and 65 °C. The results indicated that the HVEF, especially at higher voltage, has a high remarkable capability to remove metal ions and pigments from soybean oil than the IBM. The highest removal capacity for metal and pigments of soybean oil bleaching were obtained in the order of HVEF-20 kV > HVEF-10 kV > IBM.     

Effects of ultraviolet (UV) exclusion on the seasonal concentration of photosynthetic and UV-screening pigments in Scots pine needles

The effects of ultraviolet (UV) radiation on the photosynthetic and UV‐screening pigments in needles of Scots pine (Pinus sylvestris L.) saplings were studied in a UV‐exclusion field chamber experiment in northern Finland (67°N) during 2001–2002. The chambers held filters that excluded both UVB and UVA, only UVB, transmitted all UV, or lacked filters. Analyses of control needles (no filter and polyethene filter) showed that the first changes to occur in spring (end of April) was an abrupt increase in the epoxidation state (EPS) of the xanthophyll cycle pigments, likely in relation with the beginning of the photosynthetic activity. The concentration of chlorophyll, lutein, neoxanthin, α‐carotene, β‐carotene, and the size of the xanthophyll cycle pool (violaxanthin+antheraxanthin+zeaxanthin=VAZ) changed only later when needles reached their summer photosynthesis state. Exclusion of UV radiation significantly affected the xanthophyll cycle but not the other photosynthetic pigments analysed. Interestingly, the effects on xanthophylls were dependent on the sampling date. Under UVA/B‐exclusion, the EPS was increased and VAZ pool size was unchanged in April, whereas EPS remained unchanged and the VAZ pool size was reduced in May and June. The existence of two sustained and active antenna modes during winter and summer could be an explanation for the specific UV‐exclusion effect in the different season. A high‐performance liquid chromatography analysis of soluble phenolics showed that the exclusion of UVA/B radiation caused a significant effect on five compounds out of 46 studied, without affecting the concentration of the total soluble phenolics. Under UVA/B‐exclusion, the concentration of three of them (secoisolariciresinol‐glucopyranoside, two unknown) was reduced while the concentration of dicoumaroyl‐astragalin and pinosylvin monomethylether was increased compared with both controls separately. In general, the exclusion of UVA/B caused a stronger effect than the exclusion of UVB on both photosynthetic and UV screening pigments. The effects of UV radiation on xanthophyll cycle pigments were season‐specific and detectable only under stressful spring conditions (freezing temperatures and high irradiance due to snow reflection). The effect on the xanthophyll cycle could be a direct consequence of UV treatments, or an indirect consequence of the changed flavonoid composition, or a combination of both.     

Formation of water-solubleMonascus red pigments by biological and semi-synthetic processes

Summary New water-soluble red pigments were produced byMonascus sp. in a chemically defined fermentation medium containing glutamate as nitrogen source. They were isolated and characterized as glutamate derivatives of the well-known orangeMonascus pigments (monascorubrin and rubropunctatin). The new pigments have several advantages over the known redMonascus pigments (rubropunctamine and monascorubramine) including very high water-solubility, higher absorption coefficient, and greater resistance to decoloration by light. Adding glutamate, glycine or leucine to a resting-cell system led to the formation of specific water-soluble red pigments corresponding to the exogenous amino acid. The water-soluble red pigments produced by resting-cells have retention times identical to those of the corresponding red derivatives made chemically from the orange pigments in methanol-phosphate buffer at pH 7. The hydrophobicities of the amino acid sources correspond to the HPLC retention times of the red pigments derived from them.     

Production of water-soluble yellow pigments via high glucose stress fermentation of Monascus ruber CGMCC 10910

Monascus pigments are secondary metabolites of Monascus species and are mainly composed of yellow pigments, orange pigments and red pigments. In this study, a larger proportion of Monascus yellow pigments could be obtained through the selection of the carbon source. Hydrophilic yellow pigments can be largely produced extracellularly by Monascus ruber CGMCC 10910 under conditions of high glucose fermentation with low oxidoreduction potential (ORP). However, keeping high glucose levels later in the culture causes translation or a reduction of yellow pigment. We presume that the mechanism behind this phenomenon may be attributed to the redox level of the culture broth and the high glucose stress reaction of M. ruber CGMCC 10910 during high glucose fermentation. These yellow pigments were produced via high glucose bio-fermentation without citrinin. Therefore, these pigments can act as natural pigments for applications as food additives.

     

Pigment signatures associated with an anoxic coastal zone: Bahia Concepcion, Gulf of California

Bahia Concepcion is a coastal lagoon that has bottom anoxic conditions and high pigment concentrations during the summer. The phytoplankton responsible for this pigment increase is enigmatic, therefore we sampled the lagoon to analyze the pigment with a C8-HPLC system to look for signatures of phytoplankton groups. Analysis reveals a low pigment concentration in the mixed layer with a higher concentration of zeaxanthin and increasing values of chlorophyll a, peridinin, and fucoxanthin below, which peaked at the depth where oxygen dramatically decreases and H(2)S increases. Below this depth, a high pigmentation was recorded and the most important signatures were six chlorophyll-like pigments that eluted between the fucoxanthin and the chlorophyll a, and one carotenoid that eluted just after the chlorophyll a. Spectral characteristics of these last pigments are very similar to pigments present in the Chlorobiales group. These results suggest that cyanobacteria, diatoms, and dinoflagellates are responsible for the chlorophyll a increases, though in highly pigmented samples, anoxygenic phototrophic bacteria are probably the main contributors to the increase in pigments.     

Physical and chemical cues influencing the oviposition behaviour of Aphidius ervi

The oviposition behaviour of the aphid parasitoid Aphidius ervi Haliday is influenced by both chemical and physical cues. Oviposition attack responses were elicited by paint pigments sealed into the tip of a glass capillary tube. Parasitoids reacted to yellow pigments with repeated oviposition attack responses, but they did not react to green pigments. The spectrum of reflected light from the yellow pigments was very similar to that from the `green' natural host Acyrthosiphon pisum (Harris), with a high proportion of the total radiation energy being emitted in the yellow-orange wavebands (580–660 nm). Pea aphid cornicle secretion also elicited oviposition attack responses, which were not exclusively induced by its pale yellow-green colour. In fact, the oviposition attack response to capillary tips coated with cornicle secretion remained evident under red light conditions, which, in contrast, nearly completely suppressed the response to yellow pigments. Chemical compounds from cornicle secretion do not appear to be involved in parasitoid orientation, even though they stimulate intense oviposition attack responses. Olfactometer experiments showed that the putative kairomone involved acts only at very short range or on contact. Host exuviae, which also elicited strong and persistent oviposition reactions from A. ervi females, appear to be a good alternative source of ovipositional kairomone(s). This work confirms the existence of an aphid cuticular kairomone.     

Reduced photoinhibition of a phycocyanin-deficient mutant of Synechocystis PCC 6714

The effects on photoinhibition of light-harvesting pigments in microalgal cells were examined using the wild type and a phycocyanin- deficient mutant (PD-1) of Synechosystis PCC 6714. Mutant PD-1 showed higher resistance to high light than the wild type in terms of the decline of photosynthetic activity at any light intensity and with various cell densities. This suggests that the loss of productivity induced by high light intensity would be improved by reducing the content of light-harvesting pigments. This revised version was published online in June 2006 with corrections to the Cover Date.     

BANYULS, a novel negative regulator of flavonoid biosynthesis in the Arabidopsis seed coat

A mutant of Arabidopsis that accumulates a high level of red pigments within the seed coat has been isolated from a population of T-DNA-transformed plants. Genetic analysis revealed that the mutation is recessive and affects maternal seed tissues only. Due to the color of the immature seeds, this mutation was named banyuls ( ban ). Pigments accumulated continuously from early seed development to the desiccation stage in the seed coat of the mutant. The phenotype of the double mutant banyuls/transparent testa confirmed the flavonoid nature of the pigments and enabled assignment of the regulatory TT ( Transparent Testa ) genes to two groups according to their epistatic relationship to ban . The flavonoid content of germinated ban and wild-type seedlings was similar. Plants harbouring the ban mutation had a normal formation of trichomes and root hairs and were not affected in their responses to light. The seeds of ban plants exhibited reduced germination compared to wild-type which may be a direct consequence of the high level of pigments. These results suggest that BANYULS functions as a negative regulator of flavonoid biosynthesis that prevents accumulation of pigments in the seed coat during early embryogenesis in Arabidopsis .