铜、镉胁迫下施硫肥和有机肥对冬小麦碳氮运转的影响

采用盆栽试验,研究了铜、镉胁迫条件下施硫和有机肥对冬小麦碳氮运转的影响。结果表明,与各自对照相比,铜、镉胁迫下低施硫和有机肥的处理增加了小麦叶片、茎鞘、颖壳穗轴等营养器官花前贮藏物质、氮素的再运转量和运转率以及营养器官花前贮藏物质、氮素的总再运转量和总运转率,高施硫和有机肥的铜、镉处理则规律性不明显。在铜、镉胁迫条件下,施用硫肥和有机肥处理增加了小麦成熟期籽粒重和花后光合同化物输入籽粒量以及籽粒氮素含量和花后氮素积累量。与各自对照相比,铜胁迫下施硫和有机肥的处理与镉胁迫下低施硫和有机肥的处理增加了成熟期小麦的穗数、穗粒数和千粒重,提高了籽粒产量,其中以T\-5处理增产幅度最大;镉胁迫下高施硫和有机肥的处理则变化不大。铜、镉胁迫下低施硫和有机肥的处理均增加了籽粒淀粉含量,而高施硫和有机肥的铜、镉处理则未表现出此规律。此外,铜、镉胁迫下施硫和有机肥的各处理增加了籽粒蛋白质的含量。     

重组人戊型肝炎疫苗细菌内毒素检查方法的建立

确立重组人戊型肝炎疫苗原液的细菌内毒素检查方法。供试品参照《中华人民共和国药典》(2005年版三部)热原检查法进行热原检查,结果符合规定。该供试品同时参照《中华人民共和国药典》(2005年版三部)细菌内毒素检查法要求进行试验。供试品溶液在40μg/m l浓度下,确定内毒素限值为40EU/m l。供试品在该内毒素限值下干扰试验有效,且细菌内毒素检查法符合规定。该疫苗用内毒素检查法代替热原检查法,方法可行。     

Stimulation of bacterial growth by heat-stable, norepinephrine-induced autoinducers

The ability of norepinephrine to increase the growth of Escherichia coli in a serum-based medium has previously been shown to be due to the production of an autoinducer of growth during early log phase. Seventeen Gram-negative and 6 Gram-positive clinical isolates were examined for a similar ability to respond to norepinephrine, and to synthesise autoinducer. The majority of Gram-negative strains both produced and responded to heat-stable norepinephrine-induced autoinducers of growth. Most of these autoinducers showed a high degree of cross-species activity, suggesting the existence of a novel family of Gram-negative bacterial signalling molecules. In contrast, although a number of Gram-positive strains were able to respond to norepinephrine, the majority failed to produce autoinducers in the presence of norepinephrine.     

Functionality of essential genes drives gene strand-bias in bacterial genomes

Essential genes, indispensable genes for an organism’s survival, encode functions that are considered a foundation of life. Based on those experimentally determined for 10 bacteria, we find that essential genes are more preferentially situated at the leading strand than at the lagging strand, for all the 10 genomes studied, confirming previous findings based on either smaller datasets or putatively assigned ones by homology search. Furthermore, we find that rather than all essential genes, only those with the COG functional category of information storage and process (J, K and L), and subcategories D (cell cycle control), M (cell wall biogenesis), O (posttranslational modification), C (energy production and conversion), G (carbohydrate transport and metabolism), E (amino acid transport and metabolism) and F (nucleotide transport and metabolism) are preferentially situated at the leading strand. In contrast, the strand-bias for essential genes in other COG functional subcategories is not statistically significant. These results suggest that the remarkable strand-bias of the distribution of essential genes is mainly relevant to the aforementioned functionalities, which, therefore, likely play a key role in shaping the gene strand-bias in bacterial genomes.     

Identification and experimental verification of a novel family of bacterial cyclic nucleotide-gated (bCNG) ion channels

Studies of bacterial ion channels have provided significant insights into the structure-function relationships of mechanosensitive and voltage-gated ion channels. However, to date, very few bacterial channels that respond to small molecules have been identified, cloned, and characterized. Here, we use bioinformatics to identify a novel family of bacterial cyclic nucleotide-gated (bCNG) ion channels containing a channel domain related by sequence homology to the mechanosensitive channel of small conductance (MscS). In this initial report, we clone selected members of this channel family, use electrophysiological measurements to verify their ability to directly gate in response to cyclic nucleotides, and use osmotic downshock to demonstrate their lack of mechanosensitivity. In addition to providing insight into bacterial physiology, these channels will provide researchers with a useful model system to investigate the role of ligand-gated ion channels (LGICs) in the signaling processes of higher organisms. The identification of these channels provides a foundation for structural and functional studies of LGICs that would be difficult to perform on mammalian channels. Moreover, the discovery of bCNG channels implies that bacteria have cyclic nucleotide-gated and cyclic nucleotide-modulated ion channels, which are analogous to the ion channels involved in eukaryotic secondary messenger signaling pathways.     

The bacterial effector Cif interferes with SCF ubiquitin ligase function by inhibiting deneddylation of Cullin1

Cycle inhibiting factor (Cif) is one of the effectors delivered into epithelial cells by enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC) via the type III secretion system (TTSS). Cif family proteins, which inhibit host cell-cycle progression via mechanisms not yet precisely understood, are highly conserved among EPEC, EHEC, Yersinia pseudotuberculosis, Photorhabdus luminescens and Burkholderia pseudomallei.Levels of several proteins relevant to cell-cycle progression are modulated by Cullin-RING ligases (CRLs), which in turn are activated by conjugation and deconjugation of NEDD8 to Cullins. Here we show that Cif interacts with NEDD8 and interferes with SCF (Skp1-Cullin1-F-box protein) complex ubiquitin ligase function. We found that neddylated Cullin family proteins accumulated and ubiquitination of p27 decreased in cells infected with EPEC. Consequently, Cif stabilized SCF substrates such as CyclinD1, Cdt1, and p27, and caused G1 cell-cycle arrest. Using time-lapse-imaging of fluorescent ubiquitination-based cell-cycle indicator (Fucci)-expressing cells, we were able to monitor cell-cycle progression during EPEC infection and confirmed the arrest of infected cells at G1. Our in vitro and in vivo data show that Cif-NEDD8 interaction inhibits deneddylation of Cullins, suppresses CRL activity and induces G1 arrest. We thus conclude that the bacterial effector Cif interferes with neddylation-mediated cell-cycle control.     

A spatio-temporal model to describe the spread of Salmonella within a laying flock

Salmonella is one of the major sources of toxi-infection in humans, most often because of consumption of poultry products. The main reason for this association is the presence in hen flocks of silent carriers, i.e. animals harboring Salmonella without expressing any visible symptoms. Many prophylactic means have been developed to reduce the prevalence of Salmonella carrier-state. While none allows a total reduction of the risk, synergy could result in a drastic reduction of it. Evaluating the risk by modeling would be very useful to estimate such gain in food safety. Here, we propose an individual-based model which describes the spatio-temporal spread of Salmonella within a laying flock and takes into account the host response to bacterial infection. The model includes the individual bacterial load and the animals’ ability to reduce it thanks to the immune response, i.e. maximum bacterial dose that the animals may resist without long term carriage and, when carriers, length of bacterial clearance. For model validation, we simulated the Salmonella spread under published experimental conditions. There was a good agreement between simulated and observed published data. This model will thus allow studying the effects, on the spatiotemporal distribution of the bacteria, of both mean and variability of different elements of host response.     

Quantitative in vitro assay to evaluate the capability of yeast cell wall fractions from Trichosporon mycotoxinivorans to selectively bind gram negative pathogens

Yeast cell wall fractions have been proposed to bind enteropathogenic bacteria. The aim of this study was to develop a quantitative assay by measuring the optical density as growth parameter of adhering bacteria. The exponential growth phase of adhering bacteria was determined by optical density reading and compared with the colony count (CFU/mL). A linear regression was compiled and the bacterial number bound to the yeast cell wall product could be determined. Further focus was the investigation of a yeast cell wall from strain Trichosporon mycotoxinivorans (MTV) for its ability to bind gram negative Salmonella, E. coli and Campylobacter strains and gram positive probiotic bacteria of the genera lactobacilli and bifidobacteria as well as gram positive Clostridium perfringens quantitatively. The gram negative probiotic strain E. coli Nissle 1917 was also investigated. Seven out of 10 S. Typhimurium and S. Enteritidis strains adhered to the cell wall product with an amount between 10³ and 10⁴ CFU/10 μg. Four out of 7 E. coli strains showed an average binding capability (10² CFU/10 µg) whereas 4 × 10³E. coli F4 cells bound per 10 μg yeast cell wall. E. coli 0149 K91, E. coli 0147 K89, C. jejuni and C. perfringens as well the genera lactobacilli and bifidobacteria did not bind to the yeast cell wall. E. coli Nissle 1917 was bound with 2 × 10² CFU/10 μg. These results demonstrate that cell wall from MTV can be used to differentially bind E. coli spp. and Salmonella spp. up to 8 × 10⁴ CFU/10 μg. Thus certain yeast cell walls may prevent enteric infections caused by selective bacteria. This methodical approach would be an accurate tool in the feed industry for quality control of yeast cell wall products.