Antioxidant enzyme activity in endemic Baikalean versus Palaearctic amphipods: tagma- and size-related changes

The activities of key antioxidant enzymes in two endemic Baikalean amphipod species: Pallasea cancelloides (Gerstf), Eulimnogammarus verrucosus (Gerstf) and the widely distributed Palearctic species Gammarus lacustris (Sars) were studied. This work was done to prove or disprove the hypothesis that Baikalean endemics have specifics in antioxidants system different from Palearctic species. The activities of antioxidant enzymes peroxidase, catalase and glutathione-S-transferase were measured in different sections (tagmata) of the amphipods' bodies as well as in different size groups. Well expressed tagma-related differences in peroxidase activity as well as smaller differences in catalase activity were shown in all studied species. There were no measured differences in glutathione-S-transferase activity among body sections. The existence of size-related changes in some antioxidant enzymes and the difference in such changes between Baikalean and Palearctic amphipods were noted. A significant increase in peroxidase activity with the size was found in both Baikalean species while a significant decrease in peroxidase activity was observed in the Palearctic G. lacustris. In Baikalean P. cancelloides, a significant decrease of catalase activity with the increase in age of crustaceans was noted, while in E. verrucosus no such relationship was found. In the Palearctic G. lacustris, a significant increase in catalase activity with the increase in size was noted. All species are shown to have no size-related differences in glutathione-S-transferase activity. The differences between species as well as between both different tagmata and size-classes within a particular species were estimated. It was assumed that the estimated differences in enzymes activity most likely depend on interspecific variation, rather than on conditional specifics in Lake Baikal.     

Whole genome expression profiling using DNA microarray for determining biocompatibility of polymeric surfaces

There is an ever increasing need to find surfaces that are biocompatible for applications like medical implants and microfluidics-based cell culture systems. The biocompatibility of five different surfaces with different hydrophobicity was determined using gene expression profiling as well as more conventional methods to determine biocompatibility such as cellular growth rate, morphology and the hydrophobicity of the surfaces. HeLa cells grown on polymethylmethacrylate (PMMA) or a SU-8 surface treated with HNO3-ceric ammonium nitrate (HNO3-CAN) and ethanolamine showed no differences in growth rate, morphology or gene expression profiles as compared to HeLa cells grown in cell culture flasks. Cells grown on SU-8 treated with only HNO3-CAN showed almost the same growth rate (36 +/- 1 h) and similar morphology as cells grown in cell culture flasks (32 +/- 1 h), indicating good biocompatibility. However, more than 200 genes showed different expression levels in cells grown on SU-8 treated with HNO3-CAN compared to cells grown in cell culture flasks. This shows that gene expression profiling is a simple and precise method for determining differences in cells grown on different surfaces that are otherwise difficult to find using conventional methods. It is particularly noteworthy that no correlation was found between surface hydrophobicity and biocompatibility.     

Behavioural and chemical studies of discrimination processes in the leaf-cutting ant Acromyrmex laticeps nigrosetosus (Forel, 1908).

Leaf-cutting ants live in symbiosis with a basidiomycete fungus that is exploited as a source of nutrients for ant larvae. Tests of brood transport revealed that Acromyrmex laticeps nigrosetosus workers did not discriminate a concolonial brood from an alien brood. The same result was observed with tests of fungus transport. Adult workers showed no aggressive behaviour to workers from other alien colonies (non-nestmates). There was no qualitative variation in the chemical profiles of larvae, pupae and adult workers from the different colonies. However, quantitative differences were observed between the different colonies. Hypotheses about the lack of intraspecific aggression in this subspecies of ants are discussed.     

Promiscuous malaria peptide epitope stimulates CD45Ra T cells from peripheral blood of nonexposed donors.

PBL from individuals with no history of malaria exposure, as well as cord blood lymphocytes, were tested for proliferation to T cell epitopes from the malaria circumsporozoite proteins of Plasmodium falciparum and Plasmodium vivax. Cells from many individuals proliferated in response to these peptides, but for two peptides (P. vivax317-336 and P. falciparum CS331-350) the response rate ranged from 64 to 93%, with the specific stimulation indices reaching as high as 38. The phenotype of the cells responding to PfCS331-350 was predominantly CD4+,CD8-,CD45Ra+,CD45Ro-, which was the inverse of the phenotype of the cells responding to tetanus toxoid with respect to CD45 isoforms. T cell clones from different individuals specific for PfCS331-350 were restricted by at least four different HLA-DR molecules and there was no evidence that the peptide was a "superantigen." Overlapping peptides were used to demonstrate that clones had different fine specificities although the peptide specificities of the DR4-restricted and DR11-restricted clones were similar. Although the individuals tested here have had no history of malaria exposure, these data demonstrate that they have T cells specific for malaria sequences present in high frequency that proliferate as intensely as some memory responses. Although one clone from an individual with a history of BCG vaccination did react strongly with PPD, the phenotype of these cells suggests that they are not classical memory cells for a cross-reactive recall Ag. Such cells may affect the induction or expression of malaria immunity.     

A comparative study of foot dimension between adult male and female and evaluation of foot hazards due to using of footwear

Using footwear often becomes troublesome and creates many problems. Most of these problems are associated with the wearing of ill-fitting footwear, as it leads to biomechanical imbalance and ultimately give rise to different foot problems. In the present investigation different foot problems, viz., discomfort, pain and other hazards related to the use of footwear have been evaluated and attempts have been made to study different foot dimensions of men and women that are related to the design of footwear. For the present study different foot dimensions of both right and left feet of the subjects were measured on 300 Bengalee (Indian) subjects having the age range of 20-35 years. The subjects reported that they had got discomfort, pain, blister and corn due to using different footwear. It was noted that the occurrence of these problems in right foot was greater than that in left foot. There was no significant correlation between foot troubles and type of footwear. Results also showed that there was no significant difference in most of the foot dimensions between left foot and right foot. However, significant difference (P < 0.001) in all foot dimensions was observed between male and female subjects. Correlation coefficient among different foot dimensions has also been evaluated and it was noted that foot length was highly correlated with stature and foot volume, particularly in left foot. Footwear should be made according to the foot dimensions of the user population. The database collected from the Bengalee (Indian) population may be a helpful guide for manufacturing different footwear.     

Harvesting Systems,Soil Cultivation,and Nitrogen Rate Associated with Sugarcane Yield

The adoption of mechanical harvesting of green cane gives rise to concerns as to whether systems developed under burnt cane harvesting are applicable to a green cane harvesting system. In particular, tillage, which is an integral part of the burnt cane system, may no longer be necessary, and the nitrogen fertilizer rates required may need to be replaced due to the large amounts of organic matter being returned to the soil after green cane harvesting. Mechanical harvesting is relatively new in Brazil and little is known about its effect on other sugarcane production strategies. This work aimed to evaluate sugarcane performance under not only different harvesting and cultivation systems, but also different nitrogen fertilizer rates over a 3-year period. The experimental design was a split plot with harvesting systems (burnt vs. green) as main plots, cultivation (interrow vs. no cultivation) as sub plots, and nitrogen rates as sub-sub plots. The harvesting systems produced similar sugarcane yields throughout the experimental period, which demonstrates that the harvest systems do not influence sugarcane yield. Mechanical tillage practices in interrow after harvesting had no impact on stalk yield or sugar quality, indicating no necessity for this operation in the following crop. Ratoon nitrogen fertilization promoted an increase of stalk and sugar yield, with highest yields obtained at the rate of 130 kg ha^?1 N. However, there was no interaction between harvesting system and nitrogen rate.     

UDP-apiose/UDP-xylose synthase. Subunit composition and binding studies.

The UDP-apiose/UDP-xylose synthase from cell suspension cultures of parsley has been purified 1400-fold by an improved method. The ratio of apiose to xylose formed from UDP-D-glucuronic acid (UDP-GlcUA) remained constant throughout the purification procedure. Dodecylsulfate-gel electrophoresis and sedimentation equilibrium measurements showed that this enzyme preparation is composed of two proteins with molecular weights of 65000 and 86000. The two proteins which are present in a molar ratio of about 1:0.7 to 1:0.9 could not be separated by ammonium sulfate fractionation, chromatography on DEAE-cellulose at different pH-values, and on omega-aminoalkyl-Sepharose, and by gel filtration on Acrylex P-100. Each protein is composed of two apparently identical subunits. The presence of only two different subunits was confirmed by end group analysis in which glycine was found as N-terminal amino acid for the larger and lysine for the smaller protein. Crosslinking with dimethylsuberimidate gave dimers of the identical subunits but no hybrids. Separation of the two proteins was achieved on DEAE-cellulose in the presence of urea. After dialysis only the 86000-Mr protein showed enzyme activity with no significant change in the apiose/xylose ratio. However, in the absence of the 65000-Mr protein enzyme stability was decreased drastically. By equilibrium dialysis it was found that 0.5 mol UDP-GlcUA are bound per mole of 86000-Mr protein. NAD+ alone was not bound, but in the presence of UDP it was also bound in a ratio of 0.5 mol/mol catalytic protein. Experiments in which sodium borohydride was added to the enzyme incubation gave no indication that the 4-keto intermediate is bound as a Schiff base to the enzyme. Also no evidence for epimerization at C-3 of the 4-ulose intermediate prior to ring contraction to apiose was found.     

Compositional transitions in the nuclear genomes of cold-blooded vertebrates

Summary The compositional properties of DNAs from 122 species of fishes and from 18 other coldblooded vertebrates (amphibians and reptiles) were compared with those from 10 warm-blooded vertebrates (mammals and birds) and found to be substantially different. Indeed, DNAs from cold-blooded vertebrates are characterized by much lower intermolecular compositional heterogeneities and CsCl band asymmetries, by a much wider spectrum of modal buoyant densities in CsCl, by generally lower amounts of satellites, as well as by the fact that in no case do buoyant densities reach the high values found in the GC-richest components of DNAs from warm-blooded vertebrates.In the case of fish genomes, which were more extensively studied, different orders were generally characterized by modal buoyant densities that were different in average values as well as in their ranges. In contrast, different families within any given order were more often characterized by narrow ranges of modal buoyant densities, and no difference in modal buoyant density was found within any single genus (except for the genusAphyosemion, which should be split into several genera).The compositional differences that were found among species belonging to different orders and to different families within the same order are indicative of compositional transitions, which were shown to be essentially due to directional base substitutions. These transitions were found to be independent of geological time. Moreover, the rates of directional base substitutions were found to be very variable and to reach, in some cases, extremely high values, that were even higher than those of silent substitutions in primates. The taxonomic and evolutionary implications of these findings are discussed.     

Self-diffusion of polymers in cartilage as studied by pulsed field gradient NMR

Pulsed field gradient (PFG) nuclear magnetic resonance (NMR) was used to investigate the self-diffusion behaviour of polymers in cartilage. Polyethylene glycol and dextran with different molecular weights and in different concentrations were used as model compounds to mimic the diffusion behaviour of metabolites of cartilage. The polymer self-diffusion depends extremely on the observation time: The short-time self-diffusion coefficients (diffusion time Delta approximately 15 ms) are subjected to a rather non-specific obstruction effect that depends mainly on the molecular weights of the applied polymers as well as on the water content of the cartilage. The observed self-diffusion coefficients decrease with increasing molecular weights of the polymers and with a decreasing water content of the cartilage. In contrast, the long-time self-diffusion coefficients of the polymers in cartilage (diffusion time Delta approximately 600 ms) reflect the structural properties of the tissue. Measurements at different water contents, different molecular weights of the polymers and varying observation times suggest that primarily the collagenous network of cartilage but also the entanglements of the polymer chains themselves are responsible for the observed restricted diffusion. Additionally, anomalous restricted diffusion was shown to occur already in concentrated polymer solutions.     

Artificial crossing experiments within Hypecoum sect. Hypecoum (Papaveraceae)

An artificial crossing program involving 8 annual form-series within Hypecoum sect. Hypecoum (Papaveraceae) was carried out. In most combinations between different species, hybrid inviability due to disparity between parental genomes seems to be the most important crossing barrier: chlorotic progeny was produced, or hybrids with low vitality, few and sterile flowers or dwarfishness occurred. The three form-series considered as different subspecies of H. procumbens seem rather to be separated by hybrid break-down mechanisms in advanced generations, expressed as malformation of organs, and often pronounced decrease in fertility. Crossings between populations within the same form-series usually yielded vigorous and highly fertile progeny. The breeding relationships within the group were found to be well correlated with discontinuities in morphological characteristics. The disparity between the genomes of the different form-series are probably mainly due to gene mutations and cryptic structural alterations and not to major structural polymorphisms.     

Action of inhibition by fluorodeoxyuridine and its reversal by thymidine on the nucleic acid fractions of the overground parts of photoperiodically inducedChenopodium rubrum plants

The nucleic acid fractions obtained by chromatography on MAK columns were compared in 4 variants ofChenopodium rubrum plants treated in different ways during floral induction. The first variant was normally induced to flowering. The second one was inhibited by application of FUDR to the apical bud on the third day of induction. In the third variant the inhibitory effect of FUDR was reversed by application of THY 24 h after FUDR treatment. In the fourth variant THY was applied 24 h after the termination of induction at a time when it was no longer able to reverse the inhibition of flowering. In plants treated with FUDR, a decrease in DNA and RNA synthesis was observed. After reversal of the inhibitory effect of FUDR by THY, DNA synthesis remained somewhat lower than in the control variant but RNA synthesis reached the same level as in the controls or even surpassed it. In plants to which THY was applied at a time when it was no longer possible to reverse flowering, the results obtained from different experiments were not identical. In some experiments nucleic acid synthesis remained lower than in the variant in which THY restituted flowering. In other experiments nucleic acid synthesis was fully restituted and reached the same level as in the control variant. This phenomenon is explained by the different size of the experimental plants at the beginning of the different experiments. The results are discussed with respect to the relation of the dynamics of nucleic acids to growth and development of the plants.     

Effects of donor cells on in vitro development of cloned bovine embryos

The donor cells from different individuals and with different foreign genes introduced were investigated to determine their effects on the efficiency of somatic cell nuclear transfer (SCNT). The bovine ear fibroblast from different individuals was isolated, cultured, and then transfected with foreign genes to establish the stable cell lines, which were used as donor cells for nuclear transfer. The ooeytes were obtained through ovum pick up operation. After in vitro maturation, the M II phase oocytes were selected as receptors for nuclear transfer.The reconstructed embryos were cultured in vitro and observed at 2 h, 48 h, and 7 days after transfer to assess the rate of fusion using cleaved and blastoeyst as the parameters of SCNT efficiency. The donor cells from different individuals (04036, 06081, 06088, and 06129)had no obvious effect on the fusion and cleaved rate, whereas there was significant difference in the blastocyst rate (P<0.05), and the rate was 62.3%, 37.0%, 35.1%, and 15.6%, respectively. There was no significant difference among the rate of fusion, cleaved and blastocyst in donor cells with different foreign genes (P>0.05). It was concluded that the genetic background of the donor cells could affect the effi-ciency of SCNT, while the introduction of foreign genes into the donor cells had no obvious effect on the efficiency. This study provides useful information for the SCNT and would benefit in promoting the efficiency.     

Catalytic properties of an expressed and purified higher plant type zeta-carotene desaturase from Capsicum annuum.

The zeta-carotene desaturase from Capsicum annuum (EC 1.14.99.-) was expressed in Escherichia coli, purified and characterized biochemically. The enzyme acts as a monomer with lipophilic quinones as cofactors. Km values for the substrate zeta-carotene or the intermediate neurosporene in the two-step desaturation reaction are almost identical. Product analysis showed that different lycopene isomers are formed, including substantial amounts of the all-trans form, together with 7,7',9,9'-tetracis prolycopene via the corresponding neurosporene isomers. The application of different geometric isomers as substrates revealed that the zeta-carotene desaturase has no preference for certain isomers and that the nature of the isomers formed during catalysis depends strictly on the isomeric composition of the substrate.     

大熊猫精浆蛋白的SDS-PAGE电泳研究

本试验采用SDS-PAGE对大熊猫精浆进行电泳并对电泳条带进行分析,探讨特异条带与大熊猫精液质量的关系。结果显示：大熊猫精浆SDS-PAGE电泳共分离得到12条蛋白谱带,其中6个条带为所有个体大熊猫精浆样品所共有,共有条带的相对量在不同个体之间不存在差异（P〉0．05）。试验中部分大熊猫精浆出现的特异蛋白带和共有带蛋白含量与精液质量无相关性。     

Assessing tillage disturbance on assemblages of ground beetles (Coleoptera: Carabidae) by using a range of ecological indices

Many ecological studies have used diversity indices to assess the impact of environmental disturbance. In particular, ground beetles have been advocated as a good group for assessing disturbance. Most studies on various organisms have used only one or two indices. For our study of the impact of tillage disturbance on carabid beetles in farm fields in southern Ontario, Canada, we used seven different diversity indices (richness, Shannon–Wiener, Berger–Parker, Q-statistic, Margalef, and evenness). Few studies have used deviation from diversity abundance models as a measure of disturbance; however, we use three that are applicable to our data (geometric, log-normal and log-series). The indices and models were used to test the null hypothesis that there is no change in diversity with increasing tillage disturbance, and that there is no difference in diversity with different crops or years. We were not able to reject the null hypothesis that there is any diversity difference among farms. We also found that there was no single diversity index or model that was better than any other at detecting disturbance. These results are supplemented by a meta-analysis of 45 published data sets for the same taxon but in different habitats. The meta-analysis supports the conclusions from our field research that diversity indices and models are not useful for detecting the possible effect of disturbance on assemblages of carabid beetles.     

alpha-Fucose inhibits human mixed-lymphocyte culture reactions and subsequent suppressor cell generation

Carbohydrate moieties serve as important sites of interaction for many lymphocyte activities. The potential role of saccharides in the cellular interactions involved in mitogen-, antigen-, and alloantigen-induced proliferation was investigated. Eight different monosaccharides were tested for their inhibitory potential when added to uni- and bidirectional mixed-lymphocyte culture (MLC) reaction as well as to mitogen (Con A, PHA, PWM)-stimulated cultures. Only alpha-L-fucose blocked the MLC reaction in a dose-dependent fashion while having no effect on mitogen stimulation, although antigen-specific stimulation was also blocked by fucose. Similarly alpha-L-fucose specifically inhibited the MLC-induced generation of suppressor cells. Pretreatment of the MLC responder cells with fucose dehydrogenase abolished the MLC reaction while stimulator cell pretreatment had no effect, suggesting that the recognition site of the former contained alpha-L-fucose. The generation and the effector phase of Con A-induced suppressor cells was not affected by fucose, indicating that different receptors are involved in the latter. Apparent competitive inhibition by exogenous fucose of the cell-cell interaction required for the MLC reaction suggested that this monosaccharide is an essential constituent of allogeneic recognition sites.     

Bacteriophages of Pseudomonas putida containing single-stranded canonical DNA breaks

It was shown that bacteriophage tf as well as bacteriophages phi p4/40, phi p25/42, phi p23/40 and phi p6/40, which are specific to different P. putida strains, contain the single strand breaks in their DNA. The breaks are localized in one strand of DNA molecules and are repairable with T4 DNA ligase. Bacteriophage tf has no detectable DNA homology with phi p4/40, phi p25/42, phi p23/40 and phi p6/40 bacteriophages. All the phages studied have no relation with other known Pseudomonas phages. Bacteriophages phi p4/40 and phi p25/42 share the extensive DNA homology.     

Two types of triplicated alpha-globin loci in humans.

DNA from healthy Malaysian newborns was studied on gene maps after digestion with different restriction endonucleases. Of 65 newborns, two were found to be carriers of two different variants of triplicated alpha-globin loci. In variant no. 1, found in an Malay, the three alpha-globin genes are in an elongated DNA fragment on digestion with Eco RI and Bam HI. The third alpha-globin gene was found in a additional 3.7-kb fragment on digestion with Hpa I, Bgl II and Hind III. In variant no. 2, a new type of triplicated alpha-globin loci, found in a Chinese, the three alpha-globin genes reside in an elongated DNA fragment longer than that of variant no. 1 on digestion with Eco RI and Bam HI. The third alpha-globin gene was found in an additional 4.2-kb fragment on digestion with Hpa I and Hind III. Digestion of this variant DNA with Bg1 II produced an abnormal 16.7-kb fragment in addition to the normal 7.0-kb Bgl-II fragment. The locations of the restriction sites in the two types of triplicated alpha-globin loci are compatible with a mechanism of unequal crossing over following two different modes of misalignment.     

不同低温胁迫对烟草愈伤组织抗氰交替途径诱导和交替氧化酶表达影响的比较

比较了不同低温（14℃和4℃）胁迫对烟草（Nicotiana rustica L.）愈伤组织抗氰交替途径诱导和交替氧化酶表达的影响。结果显示,不同低温胁迫处理能显著诱导烟草愈伤组织交替途径容量和实际运行的增加,且都呈现出基本相同的变化模式：在胁迫的初期（1～3 d）持续增加,在3 d时达到最高,而后下降到一个相对恒定的水平。但交替途径容量增加的幅度与温度下降的程度密切相关,而交替途径实际运行量的诱导程度在不同低温胁迫下的差异却很小。表明交替途径容量和实际运行对低温胁迫的响应是不同的。免疫印迹分析结果表明：低温胁迫明显诱导了交替氧化酶总蛋白的增加,且其随低温胁迫进程的变化与交替途径容量的变化基本一致;而对交替氧化酶单体与二聚体在低温胁迫下的含量变化检测结果则显示,烟草愈伤组织中交替氧化酶主要以二聚体形式存在,且这一存在形式并不随低温胁迫程度的加深而发生改变。两种形式的交替氧化酶蛋白含量都能被低温胁迫诱导增加,但其单体水平在两种不同的低温胁迫下并无明显差别,而4℃低温胁迫诱导的二聚体交替氧化酶蛋白含量明显高于14℃。表明不同程度低温对抗氰交替途径发生的不同影响主要是由于对交替氧化酶蛋白二聚体形式的不同诱导程度所致;而高活性的交替氧化酶单体形式则不因低温胁迫程度的加重而被明显诱导升高,使得抗氰交替途径的运行程度在两种不同的低温胁迫处理条件下无显著差异。     

Morphological and anatomical studies inTribulus terrestris I. Phyllotaxis and branching

Phyllotaxis and branching system inTribulus terrestris were studied. Besides three nodal types already known in the species, a new type was found. The four nodal types were designated as A (with one leaf and no flower), B (with one leaf and one flower), C (newly found type, with two leaves of different sizes and no flower) and D (with two leaves of different sizes and one flower). A-type nodes are found only in basal parts of shoots, while distal parts have exclusively D-type nodes. Between the nodes of the two types there is always one node of B-type. Occurrence of C-type is rare and restricted to the first node of the lateral branch. The first leaf of the lateral branch superposes to the leaves of the mother axis at maturity, but it is situated at first more or less laterally. On the basis of anatomy, as well as gross morphology, the branching system at B- and D-type nodes was interpreted as sympodial, the main axis ending in a flower. With regard to the manner of overlapping of sepal margins, right- and left-handed calyces were recognized. The handedness always corresponds to the direction to which the peduncle is inclined.