浮游动物诱发藻类群体的形成

从研究蓝藻水华形成机理的需要出发,综述了浮游动物的牧食压力对藻类群体形成的诱发作用。指出诱发藻类群体形成的化合物来自牧食性浮游动物对藻类的有效牧食,是藻类群体形成的重要原因之一,而这些诱发性的化合物并不是有关生物体的组成成分,是种间相互作用的结果。藻类群体的形成方式有源于一个母细胞的分裂和业已存在的单细胞的聚合两种方式,栅藻的诱发性群体可能是来自一个母细胞的分裂,而在其它藻类的诱发性群体形成如铜绿微囊藻则可能是业已存在的单细胞的聚合。由于藻类形成群体后能显著降低浮游动物对其牧食速率,因此,这种诱发性群体形成的现象,可以解释为藻类对变化的牧食压力的一种有效的反牧食防御策略,也是两者协同进化的结果。浮游动物对藻类群体形成的重要作用,在研究模拟蓝藻群体及水华形成值得借鉴应用。作者还提出推测,水华蓝藻的群体形成,可能就是在富营养化条件下藻类快速生长,加上浮游动物的牧食压力共同作用下联合驱动的结果,而这种群体形成很可能在积累到一定程度后,结合特定的气象水文等理化因子,就会聚集于水表“爆发”出肉眼可见的水华。因此,开展浮游动物牧食作用对水华蓝藻早期群体形成诱发效应的研究不仅能加深对水华形成的全面认识,而且对于进一步认识藻类的诱发性反牧食防御适应机制、揭示生态系统中生物之间的复杂关系也具有重要的理论意义。     

Seed germination and seedling survival in a colony of the common cormorant,Phalacrocorax carbo

Germination and seedling survival of some woody plants were examined in a colony of the common cormorant at Unoyama, central Japan. The germination ofQuercus serrata andPinus densiflora was less successful in the colony than outside the colony. The percentage of germination success was negatively correlated with the amount of cormorant faeces scattered on the ground and also with the soil water conent. Most of the germinated seedlings inside of the colony died with symptoms such as necrosis spreading from the edge of leaves. Saplings ofQ. serrata also tended to die more in the colony than outside the colony. The survival rate ofQuercus glauca seedlings with scattered faeces on their leaves and/or on the the surrounding soil was significantly lower than the rate of those free of faeces. These results suggest that in the cormorant colony, germination of seeds and seedling survival are greatly inhibited due to both the direct and indirect effects of cormorant faeces.     

Relationship between stability of folding intermediates and amyloid formation for the yeast prion Ure2p: a quantitative analysis of the effects of pH and buffer system

The dimeric yeast protein Ure2 shows prion-like behaviour in vivo and forms amyloid fibrils in vitro. A dimeric intermediate is populated transiently during refolding and is apparently stabilized at lower pH, conditions suggested to favour Ure2 fibril formation. Here we present a quantitative analysis of the effect of pH on the thermodynamic stability of Ure2 in Tris and phosphate buffers over a 100-fold protein concentration range. We find that equilibrium denaturation is best described by a three-state model via a dimeric intermediate, even under conditions where the transition appears two-state by multiple structural probes. The free energy for complete unfolding and dissociation of Ure2 is up to 50 kcal mol(-1). Of this, at least 20 kcal mol(-1) is contributed by inter-subunit interactions. Hence the native dimer and dimeric intermediate are significantly more stable than either of their monomeric counterparts. The previously observed kinetic unfolding intermediate is suggested to represent the dissociated native-like monomer. The native state is stabilized with respect to the dimeric intermediate at higher pH and in Tris buffer, without significantly affecting the dissociation equilibrium. The effects of pH, buffer, protein concentration and temperature on the kinetics of amyloid formation were quantified by monitoring thioflavin T fluorescence. The lag time decreases with increasing protein concentration and fibril formation shows pseudo-first order kinetics, consistent with a nucleated assembly mechanism. In Tris buffer the lag time is increased, suggesting that stabilization of the native state disfavours amyloid nucleation.     

Product formation kinetics in genetically modified <Emphasis Type="Italic">E. coli </Emphasis>bacteria: inclusion body formation

A data-driven model is presented that can serve two important purposes. First, the specific growth rate and the specific product formation rate are determined as a function of time and thus the dependency of the specific product formation rate from the specific biomass growth rate. The results appear in form of trained artificial neural networks from which concrete values can easily be computed. The second purpose is using these results for online estimation of current values for the most important state variables of the fermentation process. One only needs online data of the total carbon dioxide production rate (tCPR) produced and an initial value x of the biomass, i.e., the size of the inoculum, for model evaluation. Hence, given the inoculum size and online values of tCPR, the model can directly be employed as a softsensor for the actual value of the biomass, the product mass as well as the specific biomass growth rate and the specific product formation rate. In this paper the method is applied to fermentation experiments on the laboratory scale with an E. coli strain producing a recombinant protein that appears in form of inclusion bodies within the cells’ cytoplasm.     

Survival of Campylobacter jejuni and Escherichia coli in groundwater during prolonged starvation at low temperatures

AIMS: To evaluate the survival of Campylobacter jejuni relative to that of Escherichia coli in groundwater microcosms varying in nutrient composition. METHODS AND RESULTS: Studies were conducted in groundwater and deionized water incubated for up to 470 days at 4 degrees C. Samples were taken for culturable and total cell counts, nutrient and molecular analysis. Die-off in groundwater microcosms was between 2.5 and 13 times faster for C. jejuni than for E. coli. Campylobacter jejuni had the lowest decay rate and longest culturability in microcosms with higher dissolved organic carbon (4 mg l(-1)). Escherichia coli survival was the greatest when the total dissolved nitrogen (12.0 mg l(-1)) was high. The transition of C. jejuni to the coccoid stage was independent of culturability. CONCLUSION: The differences in the duration of survival and response to water nutrient composition between the two organisms suggest that E. coli may be present in the waters much longer and respond to water composition much differently than C. jejuni. SIGNIFICANCE AND IMPACT OF THE STUDY: The data from these studies would aid in the evaluation of the utility of E. coli as an indicator of C. jejuni. This study also provided new information about the effect of nutrient composition on C. jejuni viability.     

Effects of LPA1 and LPA6 on the regulation of colony formation activity in colon cancer cells treated with anticancer drugs

Lysophosphatidic acid (LPA) is a simple physiological lipid and exhibits a variety of cellular responses via the activation of G protein-coupled transmembrane LPA receptors (LPA receptor-1 (LPA₁) to LPA₆). The aim of our study was to investigate effects of LPA receptors on soft agar colony formation in colon cancer cells treated with anticancer drugs. DLD1 cells were treated with fluorouracil (5-FU) or cisplatin (CDDP) for at least six months (DLD-5FU and DLD-CDDP cells, respectively). LPAR1 gene expression was markedly elevated in DLD-5FU cells. In contrast, DLD-CDDP cells showed the high expression of LPAR6 gene. In colony formation assay, DLD-5FU cells formed markedly large-sized colonies, while no colony formation was observed in DLD1 and DLD-CDDP cells. The large-sized colonies formed in DLD-5FU cells were suppressed by LPA₁ knockdown. In contrast, LPA₆ knockdown increased the size of colonies. In addition, DLD-5FU cells were further treated with CDDP for three months (DLD-C-F cells). DLD-CDDP cells were also treated with 5-FU (DLD-F-C cells). DLD-C-F cells formed large-sized colonies, but not DLD-F-C cells, correlating with LPAR1 and LPAR6 gene expression levels. These results suggest that LPA₁ and LPA₆ may regulate the colony formation activity in DLD1 cells treated with anticancer drugs.     

Socioeconomic measures influence survival in osteosarcoma: an analysis of the National Cancer Data Base

BackgroundWhile previous studies have identified low socioeconomic status as a risk factor for metastatic disease in patients with high-grade osteosarcoma, the influence of socioeconomic status on overall survival remains unclear. The present study aims to investigate the relationship between survival and socioeconomic status in patients with high-grade conventional osteosarcoma.MethodsThe National Cancer Data Base (NCDB) was queried from 1998-2012 to identify all patients <40 years of age with a diagnosis of high-grade conventional osteosarcoma. A total of 3,503 patients were identified that met inclusion and exclusion criteria. Univariate relationships were investigated using Kaplan-Meier survival analysis and associated log-rank tests in order to determine patient, socioeconomic, tumor, and treatment variables associated with overall survival. Multivariate analysis was performed to determine independent predictors of survival.ResultsIn order of decreasing magnitude, metastatic disease (Hazard Ratio [HR] 3.28, 95% Confidence Interval [CI] 2.82-3.82), primary site in the pelvis or spine (HR 2.15, 95% CI 1.79-2.59), positive surgical margins (HR 1.82, 95% CI 1.46-2.27), tumor size >8 cm (HR 1.47, 95% CI 1.24-1.74), age ≥18 years (HR 1.30, 95% CI 1.14-1.48), lowest quartile of composite socioeconomic status (HR 1.23, 95% CI 1.02-1.51), and Medicaid insurance (HR 1.18, 95% CI 1.02-1.38) were predictors of decreased survival at 5 years.ConclusionTreating providers should be aware that some of their patients may have challenges unrelated to their diagnosis that make timely presentation, adherence to treatment, and continued close surveillance difficult. This investigation suggests that socioeconomic variables influence overall survival for osteosarcoma in the United States, although not as dramatically as established tumor- and treatment-related risk factors.     

Changes in the genetic level and the effects of age at first calving and milk production on survival during the first lactation over the last 25 years

Survival during the first year after first calving was investigated over the last 25 years, 1989–2013, as well as how the association of survival with season of calving, age at first calving (AFC) and within-herd production level has changed over that period. The data set contained 1 108 745 Dutch black-and-white cows in 2185 herds. Linear models were used to estimate (1) effect of year and season and their interaction and (2) effect of AFC, within-herd production level, and 5-year intervals and their two-way interactions, and the genetic trend. All models contained AFC and percentage of Holstein Friesian as a fixed effect, and herd-year-season, sire and maternal grandsire as random effects. Survival and functional survival were analysed. Functional survival was defined as survival adjusted for within-herd production level. Survival rate increased by 8% up to 92% in the last 25 years. When accounting for pedigree, survival showed no improvement up to 1999, but improved since then. Genetically, survival increased 3% to 4% but functional survival did not increase over the 25 years. We found an interesting difference between the genetic trends for survival and functional survival for bulls born between 1985 and 1999, where the trend for survival was still increasing, but was negative for functional survival. Since 1999, genetic trend picked up again for both survival and functional survival. AFC, season of calving and within-herd production level affected survival. Survival rate decreased 0.6%/month for survival and 1.5% for functional survival between AFC of 24 and 32 months. Calving in summer resulted in 2.0% higher survival than calving in winter. Within herd, low-producing cows had a lower survival rate than high-producing cows. However, these effects became less important during the recent years. Based on survival optimum AFC is around 24 months, but based on functional survival it is better to have an AFC<24 months. Overall, survival rate of heifers has improved considerably in the past 25 years, initially due to the focus on a high milk production. More recently, the importance of a high milk production has been reduced towards attention for functional survival.     

Trade‐offs in the evolution of bumblebee colony and body size: a comparative analysis

Trade‐offs between life‐history traits – such as fecundity and survival – have been demonstrated in several studies. In eusocial insects, the number of organisms and their body sizes can affect the fitness of the colony. Large‐than‐average body sizes as well as more individuals can improve a colony's thermoregulation, foraging efficiency, and fecundity. However, in bumblebees, large colonies and large body sizes depend largely on high temperatures and a large amount of food resources. Bumblebee taxa can be found in temperate and tropical regions of the world and differ markedly in their colony sizes and body sizes. Variation in colony size and body size may be explained by the costs and benefits associated with the evolutionary history of each species in a particular environment. In this study, we explored the effect of temperature and precipitation (the latter was used as an indirect indicator of food availability) on the colony and body size of twenty‐one bumblebee taxa. A comparative analysis controlling for phylogenetic effects as well as for the body size of queens, workers, and males in bumblebee taxa from temperate and tropical regions indicated that both temperature and precipitation affect colony and body size. We found a negative association between colony size and the rainiest trimester, and a positive association between the colony size and the warmest month of the year. In addition, male bumblebees tend to evolve larger body sizes in places where the rain occurs mostly in the summer and the overall temperature is warmer. Moreover, we found a negative relationship between colony size and body sizes of queens, workers, and males, suggesting potential trade‐offs in the evolution of bumblebee colony and body size.     

Assessment of the role of a terrestrial isopod in the survival of a genetically modified pseudomonad and its detection using the polymerase chain reaction

Abstract The effects of a terrestrial isopod, Porcellio scaber , on the survival of a genetically modified pseudomonad were studied. Pseudomonas fluorescens KTG was inoculated onto ash leaf litter and supplied to populations of P. scaber . Plate counts were lower in fresh faeces than the ash leaf litter for P. fluorescens KTG, and higher counts were detected in the faeces for the total bacterial population. When faeces were aged by incubation for up to 7 days at 15–17°C, plate counts for P. fluorescens KTG increased during the first day to a level similar to those in the corresponding ash leaf litter, and remained relatively constant thereafter. The total bacterial population in the faeces continued to increase steadily over the 7 days, whilst remaining at a constant level in the ash leaf litter during the same period. Counts of bacteria in faecal material showed that P. fluorescens KTG was present for 6 days after the isopods had fed on inoculated litter although transit times of food through the gut were as little as 5 h. The implications for GEMMO dispersal of bacterial retention in the gut is considered. The polymerase chain reaction was utilised in the detection of the inserted DNA. Positive amplification of the inserted DNA sequence of P. fluorescens KTG was achieved in ash leaf litter, fresh faeces, and faeces from animal which were supplied uninoculated litter for one day after feeding on the inoculated litter. However, plate counts were more sensitive than the polymerase chain reaction in detecting P. fluorescens KTG in the faeces. Our findings suggest that when the GEMMO is ingested by the woodlouse it can survive within the guts and faeces. This has implications for risk assessment of genetically modified bacteria in terrestrial environments.     

Starvation-induced expression of SspA and SspB: the effects of a null mutation in sspA on Escherichia coli protein synthesis and survival during growth and prolonged starvation

Maxicell labelling and two-dimensional gel electro-phoresis (2-D PAGE) have identified the proteins encoded by sspA and sspB (SspA, SspB) as proteins D27.1 and A25.8, respectively, in the Escherichia coli gene-protein database. SspA expression increases with decreasing growth rate and is induced by glucose, nitrogen, phosphate or amino acid starvation. The promoter, P_ssp, is similar to gearbox promoters. Inactivation of SspA (sspA::neo) blocks sspB expression. [³⁵S]-methionine-labelled proteins synthesized during growth and during stationary phase are different in δsspA strains compared to sspA strains. This difference is enhanced during extended stationary phase (24–72 h). Long-term (10 d) viability of arginine-starved isogenic strains shows that sspA cultures remain viable significantly longer than δsspA mutants. 2-D PAGE of proteins expressed during exponential growth shows that expression of at least 11 proteins is altered in δsspA strains. A functional relA gene is required for sspA to affect protein synthesis.     

A real-time analysis of growth cone–target cell interactions during the formation of stable contacts between hippocampal neurons in culture

Mechanisms of cell-cell recognition and structural changes of growth cones (g.c.) and target membranes during contact formation are poorly understood. To examine these issues, we obtained a high magnification, realtime record of stale contact formation in cultured cells from the hippocampal CA1 area in the newborn rat. We used differential interference contrast (DIC) optics coupled to a video microscope for periods of over 24 h of continuous time-lapse recording. Our goal was to observe the sequential changes exhibited by afferent and target cells as they form a stable contact. Understanding the process of how stable contacts are made is important because such contacts are the first step in synapse formation. Four principal observations emerged from our study: (1) The target cell was receptive to a contact on a specific patch on its surface defined by the presence of lamellae and filopodia. This specific patch (named target site) was invariably present on the target cell surface before the time the growth cone arrived. (2) Stable adhesion between filopodia on the two cells initiated events leading to cell–cell contact formation. Specifically, the remaining filopodia on the growth cone and target cell were redirected toward the adhering filopodia, and the growth cone size decreased dramatically. (3) The axonal process then grew at a significantly accelerated rate (up to 50 times its baseline growth rate). (4) In addition, a number of observations were obtained on axonal turns towards the target cell, induction of target sites, and architectural remodelling of cells after the formation of a new contact. Our findings indicate that in this neuronal system, filopodia are the means used by cells to interact at stages prior to and during contact formation. We speculate that the molecules involved in cell recognition and the machinery that initiates contact formation are embedded in the fine structure of filopodia. Finally, our results provide possible clues as to some of the stages that may be involved in synapse formation in the mammalian central nervous system. © 1992 John Wiley & Sons, Inc.     

Location matters: survival of artificial nests is higher in small grassland patches and near the patch edge

Nest survival is an important part of breeding success in grassland ecosystems, and the location of nests can determine vulnerability to different predators. We conducted an experiment with artificial nests to evaluate jointly the predation rate on nests at different spatial scales (landscape, patch and tussock) and the relative abundance of potential nest predators (small mammals and birds) in a temperate grassland area. In November 2014 and 2016, we installed 288 artificial nests in Common Pampas Grass Cortaderia selloana grasslands in the southeastern Pampas region, Argentina. The nests were placed in two 10-ha plots in a continuous grassland patch (c. 900 ha) within a reserve and in two small grassland patches (1.5 and 1.8 ha) in an agricultural matrix (landscape-scale), at the patch edge and inside the patches (patch-scale), and at two heights within the tussock grass (tussock-scale). In 2016, we also conducted live trapping of small mammals and surveyed birds along strip transects at the sampling sites. Nests located in patches within an agricultural matrix and near the edge had greater relative survival than those set in the reserve and inside the patches, respectively. This might be explained by the lower relative abundance of small mammals that we found outside the reserve. Artificial nest survival values recorded at the landscape-scale contrasted with those previously observed for natural nests. Our results could be partly explained by differences in nest density between agro-patches and those within the reserve. Future studies could also evaluate the role of parental nest defence on nest survival.     

Vesicle formation in the membrane of onion cells (Allium cepa) during rapid osmotic dehydration

Background and Aims

Optimization of osmotic dehydration in different plant cells has been investigated through the variation of parameters such as the nature of the sugar used, the concentration of osmotic solutions and the processing time. In micro-organisms such as the yeast, Saccharomyces cerevisiae, the exposure of a cell to a slow increase in osmotic pressure preserves cell viability after rehydration, while sudden dehydration involves a lower rate of cell viability, which could be due to membrane vesiculation. The aim of this work is to study cytoplasmic vesicle formation in onion epidermal cells (Allium cepa) as a function of the kinetics of osmotic pressure variation in the external medium.

Methods

Onion epidermal cells were submitted either to an osmotic shock or to a progressive osmotic shift from an osmotic pressure of 2 to 24 MPa to induce plasmolysis. After 30 min in the treatment solution, deplasmolysis was carried out. Cells were observed by microscopy during the whole cycle of dehydration–rehydration.

Key Results

The application of an osmotic shock to onion cells, from an initial osmotic pressure of 2 MPa to a final one of 24 MPa for <1 s, led to the formation of numerous exocytotic and osmocytic vesicles visualized through light and confocal microscopy. In contrast, after application of a progressive osmotic shift, from an initial osmotic pressure of 2 MPa to a final one of 24 MPa for 30 min, no vesicles were observed. Additionally, the absence of Hechtian strand connections led to the bursting of vesicles in the case of the osmotic shock.

Conclusions

It is concluded that the kinetics of osmotic dehydration strongly influence vesicle formation in onion cells, and that Hechtian strand connections between protoplasts and exocytotic vesicles are a prerequisite for successful deplasmolysis. These results suggest that a decrease in the area-to-volume ratio of a cell could cause cell death following an osmotic shock.     

Sensitive liquid chromatographic method using fluorescence detection for the determination of estradiol 3- and 17-glucuronides in rat and human liver microsomal incubations: formation kinetics

We have developed a sensitive and specific HPLC-fluorescence assay for the determination of estradiol-3-glucuronide and estradiol-17-glucuronide in human and rat liver microsomal incubations. The method utilizes a mobile phase comprised of acetonitrile and 50 mM ammonium phosphate buffer (35:65, v/v) that is pumped though a phenyl column at 1 ml/min; the run time is less than 15 min. Calibration curves for both metabolites were linear over the range 20-4000 pmol. The intra- and inter-day coefficients of variation were <6%. In both rat and human liver microsomes, the formation of estradiol-3-glucuronide displayed atypical kinetics (consistent with activation), while estradiol-17-glucuronide formation was consistent with classical Michaelis-Menten kinetics. Overall, the assay described is a sensitive and reproducible method for the determination of estradiol glucuronides in liver microsomal preparations.     

Role of colanic acid exopolysaccharide in the survival of enterohaemorrhagic Escherichia coli O157:H7 in simulated gastrointestinal fluids

AIM: This study evaluated the production of colanic acid (CA) exopolysaccharide (EPS) by Escherichia coli O157:H7 in relation to the pathogen's ability to survive under acidic conditions simulating the environment in the human gastrointestinal tract. METHODS AND RESULTS: Escherichia coli O157:H7 W6-13 and its CA-deficient mutant M4020 were examined for their resistance to bile salts, and their ability to survive in simulated gastric fluid containing pepsin (pH 2.0) and simulated intestinal fluid containing pancreatin (pH 8.0). The effect of acid adaptation at pH 5.5 on the survival of E. coli O157:H7 in simulated gastric fluid was also determined. The results indicated that the survivability of M4020, under conditions simulating the environment in the human gastrointestinal tract, reduced more drastically than the viability of W6-13. The presence of bile salts had a slight effect on both types of E. coli O157:H7 cells. The loss of CA did not change the ability of M4020 to respond to acid adaptation. CONCLUSION: The EPS CA may serve as a protective barrier to E. coli O157:H7 for its survival in the human gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: The study contributes to a better understanding of the EPS affecting the ability of E. coli O157:H7 to combat acid stress.     

整合转录组数据鉴定大型真菌原基形成的潜在通路

[背景]大型真菌子实体发育特别是从菌丝体到原基转变的分子机制,目前仍不清楚.现有的研究大多集中在有限的真菌种类或环境因素上,但对在发育中起关键作用的基因提供的信息有限.[目的]研究大型真菌原基形成的分子机制.[方法]对11个物种及4个环境因子相关的转录组数据进行分析.[结果]与对照组相比,上调基因数量从白灵菇(Pleu...     

Model for prebiotic pyrophosphate formation: Condensation of precipitated orthophosphate at low temperature in the absence of condensing or phosphorylating agents

Summary The formation of pyrophosphate (PPi) by condensation of orthophosphate (Pi) at low temperature (37°C) in the absence of condensing or phosphorylating agents could have been an ancient process in chemical evolution. In the present investigation the synthesis of³²PPi from³²Pi was carried out at pH 8.0 and PPi was found in larger amounts in the presence of insoluble Pi (with calcium or manganese ions) than in its absence (with magnesium ions, or with no divalent cations present). After 10 days of incubation in the presence of precipitated calcium phosphate, about 1.6 nmol/ml of PPi was formed (0.057% yield relative to insoluble Pi). The hypothesis that the reaction is dependent on precipitated Pi was reinforced by the effect of adding dimethyl sulfoxide (2.1–9.9 M) in the presence of magnesium ions: the amount of magnesium phosphate precipitated in the presence of the organic solvent was proportional to the amount of PPi formed. The large and negative activation entropies found in aqueous media with calcium ions and in a medium containing 11.3 M dimethyl sulfoxide with magnesium ions suggest that the reaction was favored by a hydrophobic phenomenon at the surface of solid Pi. This reaction could serve as a model for prebiotic formation of PPi.     

Polygenic analysis of pattern formation in Drosophila: Specification of campaniform sensilla positions on the wing

This study was designed to measure the degree of correlation between vein formation and the specification of campaniform sensillae positions in the wing of Drosophila melanogaster. The campaniform sensillae are sensory organs placed at various locations on the wing. Those on the third longitudinal vein (L3) were the focus of this analysis. The system of polygenic modifiers of vein length is comparatively simple, as shown in whole chromosome assays of selected lines. This variability provides a sensitive method of altering vein-forming ability and of assessing correlated changes in other parts of the vein pattern. In selection lines of veinlet, sensillae were displaced toward the base of the wing as vein length decreased by distal loss of vein material. Changes in the amount of vein were, however, not directly proportional to changes in sensillae positions. The more distal sensillae were shifted the largest amount. In the mutant tilt, in which reduced L3 vein-forming competence results in subterminal gaps, distal campaniform sensillae were almost completely eliminated. The remaining sensillae were shifted toward the base of the wing where vein formation is normal. The placement of sensillae therefore appears to be sensitive to the same underlying determinants involved in vein-forming competence.