Maximin 9, a novel free thiol containing antimicrobial peptide with antimycoplasma activity from frog Bombina maxima

Amphibian skin is a rich resource of antimicrobial peptides, like maximins and maximin Hs from frog Bombina maxima. Novel cDNA clones encoding a precursor protein, which comprises a novel maximin peptide (maximin 9) and reported maximin H3, were isolated from two constructed skin cDNA libraries of B. maxima. The predicted primary structure of maximin 9 is GIGRKFLGGVKTTFRCGVKDFASKHLY-NH2. A surprising substitution is at position 16, with a free cysteine in maximin 9 rather than usual conserved glycine in other reported maximins. Maximin 9, the homodimer form and its Cys(16) to Gly(16) mutant were synthesized and their antimicrobial activities were evaluated. Unlike previously reported maximin 3, the tested bacterial and fungal strains were resistant to maximin 9, its homodimer and the Cys(16) to Gly(16) mutant (with MICs > 100 mu M). On the other hand, interestingly, while eight clinical Mollicutes strains were generally resistant to maximin 9 homodimer and its Cys(16) to Gly(16) mutant, most of them are sensitive to maximin 9 at a peptide concentration of 30 mu M, especially in the presence of dithiothreitol. These results indicate that the presence of a reactive Cys residue in maximin 9 is important for its antimycoplasma activity. The diversity of antimicrobial peptide cDNA structures encountered in B. maxima skin cDNA libraries and the antimicrobial specificity differences of the peptides may reflect well the species' adaptation to the unique microbial environments. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Avoidant Learning Ability in Free Flying Housefly (Aldrichina grahami)by Electric Shock

下载PDF阅读器研究证实:蜜蜂和果蝇具有良好的学习记忆能力.利用自主改良的研究装置对另一种具有强大生存本能的双翅目昆虫--巨尾阿丽蝇(Aldrichina grahami)在自由状态下电击同避学习能力进行研究.结果表明,巨尾阿丽蝇具有良好的学习记忆能力,因为当刺激电压范围为5V到45V时,观察到巨尾阿丽蝇有显著的回避电刺激行为,而当电压达到60V时会受到明显伤害.由此推测,巨尾阿丽蝇适合作为神经系统研究的动物模型.该实验所采用的实验范例较以往有所改进,适合作为自由状态下研究昆虫的工具.

Development of zona-free hamster ova to blastocysts in vitro

The zona pellucida (ZP) enclosing the mammalian ovum is important for its protection and for initial stages of fertilization, but the role of the ZP during embryo development is less clear. This study was designed to investigate if the hamster ZP is needed for embryo development from 1-cell to blastocyst in vitro, and to compare methods for removing the ZP. A total of 395 hamster pronucleate ova were collected 10 h post activation from superovulated, mated female hamsters. The ZP was removed from some ova using either 0.05% pronase, 0.05% trypsin or acid Tyrode's solution. To prevent ZP-free ova from sticking together, they were cultured singly in 30-50 muL drops of HECM-6 culture medium together with ZP-intact ova as controls. There was no significant difference among treatment groups in embryo development to blastocyst: 36/87 (42%) in the ZP intact group; 35/75 (47%) in the pronase-treated ZP-free group; 37/74 (50%) in the trypsin-treated ZP-free group; and 37/71 (52%) in the acid-treated ZP-free group. These results indicate that 1) the ZP is unnecessary for hamster embryo development in vitro from the pronucleate ovum stage to blastocyst; 2) none of the three ZP-removal methods was detrimental to embryo development; 3) embryos do not need to be cultured in groups during in vitro development from 1-cell to blastocyst. (C) 2000 by Elsevier Science Inc.

Serum-free culture of rhesus monkey embryonic stem cells

Previous studies have shown that the maintenance and proliferation of undifferentiated rhesus monkey embryonic stem (rES) cells requires medium supplemented with fetal bovine serum (FBS). Due to the uncharacterized composition and variation in serum nature, the present study aimed to replace the serum-containing medium with a serum-free medium in the rES cell culture. The results showed that after the initial 48-h culture in the routinely used serum-containing medium, rES cells can grow and proliferate for a prolonged period in the serum-free medium composed of DMEM supplemented with a cocktail of BSA, IGF-1, TGF-alpha, bFGF, aFGF, estradiol, and progesterone. rES cells cultured in the serum-free medium maintained high level of alkaline phosphatase activity and OCT4 level. There was no indication of differentiation as judged by the marker gene expression of all three embryonic germ layers and trophoblast. In addition, serum-free culture would not affect the passage capacity and differentiation potential of rES cells. This work will facilitate the future study of induced differentiation of rES cells and other applications.

Feeder layer- and serum-free culture of rhesus monkey embryonic stem cells

The common culture system of rhesus monkey embryonic stem (rES) cells depends largely on feeder cells and serum, which limits the research and application of rES cells. This study reports a feeder layer-free and serum-free system for culture of rES cells.

Physiological responses to capture and handling of free-ranging male Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis)

We captured free-ranging male Yangtze finless porpoises over three seasons and assayed leukocytes and serum biochemistry to investigate physiological responses to the capture and handlings. Serum thyroid hormones (THs) declined sharply in those porpoises compared with hormone variation in a captive male finless porpoise. Hypernatremia and hypokalemia were also significant in the free-ranging animals suggesting that conservation of serum sodium might be acutely vital for this freshwater subspecies. The animals captured in spring showed more significant neutrophilia and eosinopenia than those captured in autumn suggesting that they may be more affected by capture during the breeding season. Furthermore, physical examination of porpoises when out of the water was apparently stressful, particularly when they were kept out of the water for longer periods. However, an increase in circulating THs may be an adaptive response to accommodate these short-term stresses.

Relationships among the contents of total phenolics, soluble carbohydrate, and free amino acids of 15 aquatic macrophytes

In order to examine how carbon and nitrogen status of a macrophyte may affect its total phenolics (TP) production, the contents of free amino acids (FAA), soluble carbohydrate (SC) and TP were examined in leaves of seven submersed, four floating-leaved, and four emergent macrophytes. The floating-leaved and emergent macrophytes had much higher contents of SC and TP than the submersed macrophytes. The contents of FAA were not significantly different among the submersed, floating-leaved, and emergent macrophytes. Correlations among the contents of FAA, SC, and TP indicated that the production of TP was more dependent on the SC content than on the FAA content.

Free radical scavenging and antioxidant enzymes activation of polysaccharide extract from Nostoc sphaeroides

Nostoc sphaeroides Kuetzing has been used as a traditional medicine in China to treat a variety of ailments. This research identified the antioxidant activities of polysaccharide extract from Nostoc sphaeroides. The extract, which contains 46.2% carbohydrates, exhibited an effective scavenging capability on superoxide radical, hydroxyl radicals in non site-specific as well as site-specific assays, and also performed lipid peroxidation inhibition in a dose-dependent manner. Polysaccharide extract had no 1,1-diphenyl-2-picrylhydrazyl radical scavenging potential at all test concentrations. Activities of superoxide dismutase, catalase, and glutathione peroxidase in human embryo kidney 293 cells were increased effectively when Nostoc sphaeroides extract was applied. These results suggested that the use of N. sphaeroides in treating ailments may be based on the antioxidant capacities of polysaccharide composition.

Biosonar behaviour of free-ranging porpoises

Detecting objects in their paths is a fundamental perceptional function of moving organisms. Potential risks and rewards, such as prey, predators, conspecifics or non-biological obstacles, must be detected so that an animal can modify its behaviour accordingly. However, to date few studies have considered how animals in the wild focus their attention. Dolphins and porpoises are known to actively use sonar or echolocation. A newly developed miniature data logger attached to a porpoise allows for individual recording of acoustical search efforts and inspection distance based on echolocation. In this study, we analysed the biosonar behaviour of eight free-ranging finless porpoises (Neophocaena phocaenoides) and demonstrated that these animals inspect the area ahead of them before swimming silently into it. The porpoises inspected distances up to 77 in, whereas their swimming distance without using sonar was less than 20 in. The inspection distance was long enough to ensure a wide safety margin before facing real risks or rewards. Once a potential prey item was detected, porpoises adjusted their inspection distance from the remote target throughout their approach.

Echolocation signals of the free-ranging Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientialis)

This paper describes the high-frequency echolocation signals from free-ranging Yangtze finless porpoise in the Tian-e-zhou Baiji National Natural Reserve in Hubei Province, China. Signal analysis showed that the Yangtze finless porpoise clicks are typical high-frequency narrow-band (relative width of the frequency spectrum Q=6.6 &PLUSMN; 1.56, N=548) ultrasonic pulses. The peak frequencies of the typical clicks range from 87 to 145 kHz with an average of 125 &PLUSMN; 6.92 kHz. The durations range from 30 to 122 μ s with an average of 68 &PLUSMN; 14.12 μ s. The characteristics of the signals are similar to those of other members of the Phocoenidae as well as the distantly related delphinids, Cephalorhynchus spp. Comparison of these signals to those of the baiji (Lipotes vexillifer), who occupies habitat similar to that of the Yangtze finless porpoise, showed that the peak frequencies of clicks produced by the Yangtze finless porpoise are remarkably higher than those produced by the baiji. Difference in peak frequency between the two species is probably linked to the different size of prefer-red prey fish. Clear double-pulse and multi-pulse reverberation structures of clicks are noticed, and there is no indication of any low-frequency (< 70 kHz) components during the recording period. © 2005 Acoustical Society of America.

Off-axis sonar beam pattern of free-ranging finless porpoises measured by a stereo pulse event data logger

The off-axis sonar beam patterns of eight free-ranging finless porpoises were measured using attached data logger systems. The transmitted sound pressure level at each beam angle was calculated from the animal's body angle, the water surface echo level, and the swimming depth. The beam pattern of the off-axis signals between 45 and 115 (where 0 corresponds to the on-axis direction) was nearly constant. The sound pressure level of the off-axis signals reached 162 dB re 1 mPa peak-to-peak. The surface echo level received at the animal was over 140 dB, much higher than the auditory threshold level of small odontocetes. Finless porpoises are estimated to be able to receive the surface echoes of off-axis signals even at 50-m depth. Shallow water systems (less than 50-m depth) are the dominant habitat of both oceanic and freshwater populations of this species. Surface echoes may provide porpoises not only with diving depth information but also with information about surface direction and location of obstacles (including prey items) outside the on-axis sector of the sonar beam. 2005 Acoustical Society of America.