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1.
The global redox switch between aerobic and anaerobic growth in Rhodobacter sphaeroides is controlled by the RegA/RegB two-component system, in which RegB is the integral membrane histidine protein kinase, and RegA is the cytosolic response regulator. Despite the global regulatory importance of this system and its many homologues, there have been no reported examples to date of heterologous expression of full-length RegB or any histidine protein kinases. Here, we report the amplified expression of full-length functional His-tagged RegB in Escherichia coli, its purification, and characterisation of its properties. Both the membrane-bound and purified solubilised RegB protein demonstrate autophosphorylation activity, and the purified protein autophosphorylates at the same rate under both aerobic and anaerobic conditions confirming that an additional regulator is required to control/inhibit autophosphorylation. The intact protein has similar activity to previously characterised soluble forms, but is dephosphorylated more rapidly than the soluble form (half-life ca 30 minutes) demonstrating that the transmembrane segment present in the full-length RegB may be an important regulator of RegB activity. Phosphotransfer from RegB to RegA (overexpressed and purified from E. coli) by RegB is very rapid, as has been reported for the soluble domain. Dephosphorylation of active RegA by full-length RegB has a rate similar to that observed previously for soluble RegB.  相似文献   

2.
Light inhibited each step of the denitrification process in whole cells of Rhodobacter sphaeroides forma sp. denitrificans. This inhibition by light was prevented in the presence of exogenous electron donors like N,N,N,N-tetramethyl-p-phenylenediamine (TMPD) plus ascorbate or in the presence of an uncoupler (carbonyl cyanide m-chlorophenylhydrazone). Addition of myxothiazol restored the inhibition by light in uncoupled cells. Measurements of light-induced absorbance changes under these conditions showed that this inhibition is due, for the steps of reduction of nitrite to dinitrogen, to the photooxidation of cytochromes c 1 plus c 2 and not due to the photoinduced membrane potential. Moreover, the presence of oxygen inhibited almost all of the reduction of nitrate and nitrous oxide but only 70% of the reduction of nitrite to nitrous oxide. These inhibitions were overcome in the presence of TMPD plus ascorbate. This implies that the inhibition in presence of oxygen was due to a diversion of the reducing power from the denitrifying chain to the respiratory chain. It was concluded from this series of experiments that the reduction of nitrate to nitrite is inhibited when the ubiquinone pool is partly oxidized and that nitrite and nitrous oxide reductions are inhibited when cytochromes c 1 plus c 2 are oxidized by photosynthesis or respiration.Abbreviations R Rhodobacter - TMPD N,N,N,N-tetramethyl-p-phenylenediamine - HOQNO 2-n-heptyl-4-hydroxyquinoline N-oxide - CCCP carbonyl cyanide m-chlorophenylhydrazone - cytochrome c 1 cytochrome c 2 plus cytochrome c 1  相似文献   

3.
Wang YJ  Liao Q  Wang YZ  Zhu X  Li J 《Bioresource technology》2011,102(13):6902-6908
Photosynthetic bacteria (PSB), Rhodopseudomonas palustris CQK 01, were immobilized on the surface of a thin glass slide in a lab-scale flat panel photobioreactor under different flow rates and substrate concentrations. The morphology, dry weight and thickness of the mature PSB biofilms were determined to reveal the relationship between biofilm formation and hydrogen production performance. The mature biofilm formed at a low flow rate and a high substrate concentration showed a looser structure, these structures of the mature biofilm then affected the H2 production performance of the bioreactor during mature stage. The biofilm formed at a flow rate of 228 ml/h and a substrate concentration of 60 mmol/l exhibited the highest dry weight and optimally porous structure, which is beneficial not only for hydrogen removal from the biofilm but also glucose diffusion into the biofilm, thus significantly boosting the photo-hydrogen production performance.  相似文献   

4.
5.
We report on the response of reaction center (RC) from Rhodobacter sphaeroides (an archetype of membrane proteins) to the exposure at high temperature. The RCs have been solubilized in aqueous solution of the detergent N,N-dimethyldodecylamine-N-oxide (LDAO). Changes in the protein conformation have been probed by monitoring the variation in the absorbance of the bacteriochlorine cofactors and modification in the efficiency of energy transfer from tryptophans to cofactors and among the cofactors (through fluorescence measurements). The RC aggregation taking place at high temperature has been investigated by means of dynamic light scattering. Two experimental protocols have been used: (i) isothermal kinetics, in which the time evolution of RC after a sudden increase of the temperature is probed, and (ii) T-scans, in which the RCs are heated at constant rate. The analysis of the results coming from both the experiments indicates that the minimal kinetic scheme requires an equilibrium step and an irreversible process. The irreversible step is characterized by a activation energy of 205 ± 14 kJ/mol and is independent from the detergent concentration. Since the temperature dependence of the aggregation rate was found to obey to the same law, the aggregation process is unfolding-limited. On the other hand, the equilibrium process between the native and a partially unfolded conformations was found to be strongly dependent on the detergent concentration. Increasing the LDAO content from 0.025 to 0.5 wt.% decreases the melting temperature from 49 to 42 °C. This corresponds to a sizeable (22 kJ/mol at 25 °C) destabilization of the native conformation induced by the detergent. The nature of the aggregates formed by the denatured RCs depends on the temperature. For temperature below 60 °C compact aggregates are formed while at 60 °C the clusters are less dense with a scaling relation between mass and size close to that expected for diffusion-limited aggregation. The aggregate final sizes formed at different temperatures indicate the presence of an even number of proteins suggesting that these clusters are formed by aggregation of dimers.  相似文献   

6.
7.
The production of biomass, polysaccharide storage material and H2 from malate was studied in the wild-type and mutants RdcI, RdcII and RdcI/cII of Rhodobacter capsulatus. The mutants are defective in either copy I, copy II or both copies of the nitrogenase genes nifA and nifB. Stationary phase levels of biomass, polysaccharide and H2 were determined in phototrophic batch cultures grown with 30 mM of d,l-malate and either 2, 5, or 8 mM of ammonium or 7 mM of glutamate. Calculation of the amounts of malate converted into the three products revealed that, at 8 mM of ammonium and 7 mM of glutamate, malate consumption and product formation were balanced. But with decreasing ammonium concentrations malate not converted into biomass was utilized with decreasing efficiency in polysaccharide and H2 formation. This suggests formation of unknown products at the lower ammonium concentrations. Under conditions of optimal N supply, 80% of the malate not used for biomass production was converted by the wild-type and strain RdcII to H2 and CO2. Mutant RdcI exhibited slightly decreased H2 production. The double mutant did not evolve H2 but accumulated increased amounts of polysaccharide. However, the amounts of polysaccharide were lower than should be expected if all of the spare malate, not utilized by the double mutant for H2 production, was converted into storage material. This and incomplete conversion of malate into known products at low ammonium supplies suggests that polysaccharide accumulation does not compete with the process of H2 formation for malate.  相似文献   

8.
The heterotrich ciliate Mirofolliculina limnoriae occurs in an obligate association with wood-boring species of the isopod Limnoria. It attaches to the dorsal surface of the limnoriid pleotelson. Folliculinid-bearing populations of Limnoria tripunctata from the Ria Formosa, Portugal, and of Limnoria quadripunctata from Portsmouth, UK, were sampled. Up to four folliculinids were found on one individual of L. quadripunctata and up to twelve on L. tripunctata. There was no evidence that folliculinid load increased with host size. The nature of the epibiont/basibiont relationship was examined by observing animals with and without epibionts, kept individually with small sticks of wood in cell culture wells over periods of up to 15 days. Faecal pellet production and moulting were recorded. In both host species, feeding rate as measured by faecal pellet production was significantly lower in animals bearing folliculinids. Feeding rate also diminished with the onset of moulting, but returned to the original rate within a week after moulting. The estimated intermoult period for the population of L. tripunctata examined was about 25 days and for L. quadripunctata was about 32 days. However, the proportion of individuals moulting over the period of observation declined with increasing size in L. tripunctata, suggesting increasing length of intermoult period with age. As the host faces toward the blind end of its tunnel and ventilates the burrow with its pleopods, the folliculinids are well placed to intercept particulate food in the incoming water stream. There are no clear advantages for the host in the association, but the suppression of feeding and the potential of the folliculinids to hinder swimming during dispersal migrations indicate that the association should be viewed as ectoparasitism rather than mutualism or commensalism.  相似文献   

9.
Summary The objectives of this investigation were to determine the effects of oxygen partial pressure (pO2) and combined nitrogen (NH 4 + ) on rates of acetylene reduction (AR) associated with roots of intact corn, sorghum, and pearl millet plants. Soil-grown plants were carefully removed from soil and incubated hydroponically with the root system enclosed in a plastic cylinder; the tops were left exposed to ambient conditions. Oxygen concentrations around the root systems were controlled by sparging the nutrient solution with known quantities of O2 in N2. Ammonium nitrogen was added to the nutrient solution following establishment of AR rates to determine its effect on rates of N2-fixation (AR). Substantial AR rates (0.1–1.5 mol C2H4 g dry wt–1 h–1) were associated with roots exposed to 0–2% O2 (v/v) (0.0–2.02 kPa) in N2 following at 12–24 h period of exposure to the reduced oxygen tension. Root systems exposed to air failed to demonstrate AR while those exposed to 100% N2 showed lower activity than those at reduced pO2 values. Addition of NH 4 + (10–20 g N ml–1 of nutrient solution) reduced AR by 75–90% within 24 h after addition. Oxygen uptake by roots exposed to low pO2 was substantially reduced.  相似文献   

10.
In many shallow coastal areas worldwide, several species of opportunistic macroalgae (mainly Chlorophyta) have an excessive growth, as a consequence of eutrophication processes. Therefore, bloom-forming macroalgae become the dominant primary producers within these coastal systems. However, frequently the ecology and the ecophysiology of adult macroalgae have been insufficient to explain their seasonal abundances. Thus, it is essential to understand the factors that regulate the germination and growth of spores of opportunistic green macroalgae. In the present work, we assessed the effects of nutrients (N and P), salinity and light on the germination and growth of Enteromorpha spores. Overall, the results highlight the fact that, such as for adult macroalgae, spore germination and growth are adversely affected by low salinities. Growth of the spores is significantly decreased at 5 psu, while salinities of 20 and, especially of 35 psu, clearly promote the spore growth. Additionally, Enteromorpha spores seem to be particularly sensitive to PO4-P limitation and to NH4-N toxicity, which suggests a higher sensitivity to the variation of external nutrient concentrations than adult macroalgae. The present results contribute to increase the understanding about the factors that control macroalgal growth at its early phases of development. In particular, the results suggest that the growth of spores from opportunistic green macroalgae is strongly salinity-dependent. Consequently, in highly hydrodynamic systems such as most shallow estuaries, salinity variations may play a determinant role in the yearly abundances of green macroalgae, since it controls macroalgal growth from the spores to the adults.  相似文献   

11.
Plants of the seagrass Zostera noltii were cultured in the laboratory (mesocosms) for two weeks to assess the effect of above:below-ground (AG/BG) biomass ratios and light on growth, photosynthesis and chemical composition. Experimental plant units (EPUs) with different proportions between AG and BG biomass were obtained from plants of the same size (containing 6 shoots and 5 internodes) by excising 0-5 shoots. The EPUs maintained the proportions in AG/BG biomass ratios during the experiment. While growth rate was unaffected by biomass partitioning at high light, maximum growth at low light was recorded in plants with low AG/BG ratios. The production of shoots and rhizomes showed a compensatory morphological response depending on the initial AG/BG proportions regardless of the light level. While shoot production, estimated as shoot appearance rate, was high at low AG/BG ratios and minimal under high AG/BG values, rhizome production, estimated as internode appearance rate and internode elongation rate, was maximal under high AG/BG proportions and decreased towards lower AG/BG ratios. This rhizomatic response was observed for secondary rhizomes and not for primary ones. In contrast to morphological response, no significant differences were detected in maximum electron transport rates (ETRm) among the different shoots in the plant. However, mean values of ETRm in plants were affected by biomass partitioning and light. EPUs grown in low light increased the sucrose stored in shoots as the AG/BG biomass ratios decreased; however, EPUs grown at high light showed no effect of biomass partitioning on sucrose levels. In conclusion, shoots excision by experimental manipulation caused a compensatory morphological response in plants while photosynthetic performance remained almost unaffected.  相似文献   

12.
Myosin light chain 2 (MLC-2) gene was isolated and characterized from Antheraea pernyi, a well-known wild silkmoth. The isolated cDNA sequence is 905 bp in length with an open reading frame of 612 bp encoding a polypeptide of 203 amino acids. Semi-quantitative RT-PCR analysis showed that the MLC-2 gene was transcribed during four developmental stages (egg, larva, pupa, and moth), and present in all tissues tested. Alignment analysis revealed that the deduced protein sequence has over 95% identity to myosin light chain 2 of lepidopteran species, and 57–88% identity to other insect species, suggesting that insect MLC-2 proteins are highly conserved throughout evolution. The protein sequence was used to construct phylogenetic trees with other known vertebrate and invertebrate MLC-2 sequences, and the obtained trees demonstrated similar topology with the classical systematics, indicating the potential value of MLC-2 gene in phylogenetic study.  相似文献   

13.
The aim of this work was to quantify the intensity of sediment mixing induced by the gallery-diffusor (functional bioturbation group) Hediste diversicolor as a function of density, using particles tracers (luminophores). In order to assess the impact of density on sediment reworking, a 1-D model was used to obtain sediment reworking coefficients such as Db (biodiffusion-like) and r (biotransport). Densities used in this experiment corresponded to population densities observed in the sampling area (Saint-Antoine Canal, Gulf of Fos, France): 144, 288, 577, 1153 indiv/m2. At first, results showed that neither luminophore maximum burying depth nor the more marked tracer accumulation areas were influenced by density. Thus density did not seem to have any influence on size of galleries or complexity of structure. Then, density-dependent relations with Db (biodiffusion-like mixing) and r (biotransport) were highlighted with an observed process intensity rate twice as high at highest worm density. On the other hand, Db and r per capita coefficients were negatively influenced by density. Db and r per capita at highest density were equal to ∼ 20% of individual Db and r obtained at the lowest density. Finally, this study showed the importance of density which appears to be a key parameter in the functioning of the sedimentary ecosystem.  相似文献   

14.
Much effort has been expended to improve irrigation efficiency and drought tolerance of agronomic crops; however, a clear understanding of the physiological mechanisms that interact to decrease source strength and drive yield loss has not been attained. To elucidate the underlying mechanisms contributing to inhibition of net carbon assimilation under drought stress, three cultivars of Gossypium hirsutum were grown in the field under contrasting irrigation regimes during the 2012 and 2013 growing season near Camilla, Georgia, USA. Physiological measurements were conducted on three sample dates during each growing season (providing a broad range of plant water status) and included, predawn and midday leaf water potential (ΨPD and ΨMD), gross and net photosynthesis, dark respiration, photorespiration, and chlorophyll a fluorescence. End-of-season lint yield was also determined. ΨPD ranged from −0.31 to −0.95 MPa, and ΨMD ranged from −1.02 to −2.67 MPa, depending upon irrigation regime and sample date. G. hirsutum responded to water deficit by decreasing stomatal conductance, increasing photorespiration, and increasing the ratio of dark respiration to gross photosynthesis, thereby limiting PN and decreasing lint yield (lint yield declines observed during the 2012 growing season only). Conversely, even extreme water deficit, causing a 54% decline in PN, did not negatively affect actual quantum yield, maximum quantum yield, or photosynthetic electron transport. It is concluded that PN is primarily limited in drought-stressed G. hirsutum by decreased stomatal conductance, along with increases in respiratory and photorespiratory carbon losses, not inhibition or down-regulation of electron transport through photosystem II. It is further concluded that ΨPD is a reliable indicator of drought stress and the need for irrigation in field-grown cotton.  相似文献   

15.
16.
Helicobacter pylori, the main cause of chronic gastritis, plays a central role in the etiology of peptic ulcer disease and gastric cancer. In vitro studies have shown that H. pylori increases gastric epithelial cell turnover, thus increasing the risk for the development of neoplastic clones. The mechanisms by which H. pylori promotes perturbation of cell proliferation are not yet elucidated. To investigate whether products released by H. pylori in culture media interfere with cell cycle progression of human gastric epithelial cells, four cell lines (MKN 28, MKN 7, MKN 74, and AGS) were incubated in the presence of H. pylori broth culture filtrate. Cell cycle analysis showed that a H. pylori-released factor(s) significantly inhibited the G1- to S-phase progression of MKN 28 and MKN 7 cell lines, with a reversible, nonlethal mechanism, independent of the expression of VacA, CagA, and/or urease. The cell cycle inhibition occurred concomitantly with an increase in p27(KIP1) protein levels, a reduction in Rb protein phosphorylation on serine residues 807-811, and a significant decrease in cyclin E-associated cdk2 activity. In contrast, the cell cycle progression of MKN 74 and AGS cell lines was not affected by the H. pylori-released factor(s). In normal human fibroblasts, G1-phase cell accumulation was concomitant with the reduction in Rb protein phosphorylation; that, however, appeared to be dependent on p21(WAF1/CIP1) rather than on p27(KIP1) protein. A preliminary characterization showed that the molecular mass of the partially purified cell cycle inhibitory factor(s) was approximately 40 kDa. These results suggest that H. pylori releases a soluble factor(s) that may affect cell cycle progression of gastric epithelial cells through elevated levels of cdk inhibitor p27(KIP1). This factor(s) might act in vivo on noncolonized distant cells, the most proliferating cells of human gastric mucosa.  相似文献   

17.
为了给大气CO2浓度逐渐升高背景下的毛竹林适应性经营管理提供理论依据,运用开顶式气室(OTCs)模拟大气CO2浓度升高(500、700 μmol/mol)情景,以目前环境背景大气为对照,研究了Na+、Fe2+-Fe3+、Ca2+、Mg2+等矿质离子在毛竹器官中吸收、运输和分配的变化规律.结果显示,除CO2浓度700 μmoL/mol对Ca2+浓度在毛竹器官中大小排序会产生影响外,CO2浓度500、700 μmol/mol并未改变毛竹器官中Na+、Fe2+,Fe3+、Mg2+、Ca2+浓度的大小排序.CO2浓度升高对竹叶Fe2+-Fe3+和竹枝Fe2+-Fe3+、Mg2+浓度无明显影响,但对器官的其它矿质离子浓度会有不同程度的影响,竹叶Ca2+和Mg2+、竹枝Na+和Ca2+、竹秆Na+和Ca2+及Mg2+、竹根Na+和Mg2+浓度明显提高,竹叶Na+、竹秆Fe2+-Fe3+、竹根Fe2+-Fe3+和Ca2+浓度明显降低;随着CO2浓度的升高,竹叶Fe2+-Fe3+/Na+、Mg2+/Na+和Ca2+/Na+,竹枝Ca2+/Mg2+及各器官Mg2+/Fe2+-Fe3+、Ca2+/Fe2+-Fe3+均逐渐增大,而竹枝、竹秆、竹根Fe2+-Fe3+/Na+、Mg2+/Na+、Ca2+/Na+和竹叶、竹秆、竹根Ca2+/Mg2+均逐渐减小;CO2浓度升高后除竹根-竹秆Sca.Na、竹秆-竹枝SMg,Fe和竹枝-竹叶Sca,Mg明显下降外,其余的毛竹器官矿质离子向上运输系数变化平缓或明显提高.研究表明CO2浓度升高增强了毛竹立竹根部积累Na+能力和Fe2+-Fe3+、Ca2+和Mg2+的向上选择性运输能力,提高了光合器官竹叶中矿质养分元素浓度,可维持体内矿质养分元素平衡,有利于提高毛竹对高浓度CO2环境的适应能力.  相似文献   

18.
Campylobacter jejuni and Mycobacterium paratuberculosis have been implicated in the pathogenesis of Crohn's disease. The presence of bacterial metabolites in the colonic lumen causing a specific breakdown of fatty acid oxidation in colonic epithelial cells has been suggested as an initiating event in inflammatory bowel disease (IBD). l-Carnitine is a small highly polar zwitterion that plays an essential role in fatty acid oxidation and ATP generation in intestinal bioenergetic metabolism. The organic cation/carnitine transporters, OCTN1 and OCTN2, function primarily in the transport of l-carnitine and elimination of cationic drugs in the intestine. High-resolution linkage disequilibrium mapping has identified a region of about 250kb in size at 5q31 (IBD5) encompassing the OCTN1 and -2 genes, to confer susceptibility to Crohn's disease. Recently, two variants in the OCTN1 and OCTN2 genes have been shown to form a haplotype which is associated with susceptibility to Crohn's. We show that OCTN1 and OCTN2 are strongly expressed in target areas for IBD such as ileum and colon. Further, we have now identified a nine amino acid epitope shared by this functional variant of OCTN1 (Leu503Phe) (which decreases the efficiency of carnitine transport), and by C. jejuni (9 aa) and M. paratuberculosis (6 aa). The prevalence of this variant of OCTN1 (Phe503:Leu503) is 3-fold lower in unaffected individuals of Jewish origin (1:3.44) compared to unaffected individuals of non-Jewish origin (1:1). We hypothesize that a specific antibody raised to this epitope during C. jejuni or M. paratuberculosis enterocolitis would cross-react with the intestinal epithelial cell functional variant of OCTN1, an already less efficient carnitine transporter, leading to an impairment of mitochondrial beta-oxidation which may then serve as an initiating event in IBD. This impairment of l-carnitine transport by OCTN1 may respond to high-dose l-carnitine therapy.  相似文献   

19.
Lee EJ  Min HY  Joo Park H  Chung HJ  Kim S  Nam Han Y  Lee SK 《Life sciences》2004,75(23):2829-2839
Stilbenoids, including resveratrol (3,5,4'-trihydroxy-trans-stilbene) which is a naturally occurring phytoalexin abundant in grapes and several plants, have been shown to be active in inhibiting proliferation and inducing apoptosis in human cancer cell lines. Using resveratrol as the prototype, we have synthesized various analogs and evaluated their growth inhibitory effects in cultured human cancer cells. In the present study, we show that one of the stilbenoids, 3,4,5-trimethoxy-4'-bromo-cis-stilbene (BCS), was more effective than its corresponding trans-isomer and resveratrol on the inhibition of cancer cell growth. Prompted by the strong growth inhibitory activity of BCS (IC50; 0.03 microM) compared to its trans-isomer (IC50; 6.36 microM) and resveratrol (IC50; 33.0 microM) in cultured human lung cancer cells (A549), we investigated its mechanism of action. BCS induced arrest at the G2/M phase cell cycle in the early time and subsequently increased in the sub-G1 phase DNA contents in a time-dependent manner, indicating induction of apoptosis. Morphological observation with round-up shape and DNA fragmentation was also revealed the apoptotic phenomena. BCS treatment elevated the expression levels of the pro-apoptotic protein p53, the cyclin-dependent kinase inhibitor p21, and the release of cytochrome c in the cytosol. The down-regulation of checkpoint protein cyclin B1 by BCS was well correlated with the cell cycle arrest at G2/M. These data suggest the potential of BCS to serve as a cancer chemotherapeutic or chemopreventive agent by virtue of arresting the cell cycle and induction of apoptosis of human lung cancer cells.  相似文献   

20.
The influence of temperature and chlorophyll concentration on egg production, hatching success, female survival and body size of the copepod Temora longicornis was investigated in situ in the coastal waters of the Eastern English Channel. Twenty samples were collected between February and July 2003 three to four times per month. The maximum daily level of egg production of 75.5 eggs female−1 was observed in March, with minima of 10 eggs female−1 in February and July. Between February and March egg production increased with chlorophyll concentration. At the end of March a decrease in egg production corresponded to a strong increase in chlorophyll concentration that indicated a Phaeocystis-dominated bloom. After this period egg production closely followed the temporal variation in chlorophyll concentration with a maximum of 55.3 eggs female−1 in April during a second peak of chlorophyll concentration. Hatching success varied between 60% and 80% and was not influenced by chlorophyll concentration, in situ temperature or any other biological parameter considered in this study. Increases in temperature from February to July paralleled a decrease in body length and an increase in the percentage of spawning females. Mean female survival followed the variation of both temperature and chlorophyll concentration only between April and July. Temperature and chlorophyll concentration affected the reproductive parameters of T. longicornis differently. Female survival and body size were negatively correlated with temperature, while the highest chlorophyll concentrations were not always favourable for egg production. Therefore the quality of food should not be associated with chlorophyll quantity. Furthermore, the maximum values of egg production in this study are the highest recorded for T. longicornis. This study, conducted for the first time in the Eastern English Channel, showed high levels of productivity of T. longicornis despite a decrease of egg production during a Phaeocystis sp. dominated bloom in April.  相似文献   

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