Influences of cadmium on fine structure and metabolism of Hypnea musciformis (Rhodophyta, Gigartinales) cultivated in vitro

The in vitro effect of cadmium on apical segments of Hypnea musciformis was examined. Over a period of 7?days, the segments were cultivated with different concentrations of cadmium, ranging from 50 to 300???M. The samples were processed for microscopic and histochemical analysis of growth rates, content of photosynthetic pigments, and photosynthetic performance. Cadmium treatments increased cell wall thickness and the accumulation of plastoglobuli. Destruction of chloroplast internal organization was observed. Compared to controls, algae exposed to cadmium showed growth rate reduction, depigmentation, and blending in the lateral branches. The content of photosynthetic pigments, including chlorophyll a and phycobiliproteins, decreased after exposure to different concentrations of cadmium. These results agree with the decreased photosynthetic performance and relative electron transport rate observed after exposure of algae to cadmium. Taken together, these findings strongly indicate that cadmium negatively affects the architecture and metabolism of the carragenophyte H. musciformis, thus posing a threat to the economic vitality of this red macroalgae.     

Modified procedures for extraction and analysis of carrageenan applied to the red alga Hypnea musciformis

Dried powder of Hypnea musciformis was extracted with water at pH 7 after an initial short pre-treatment with cold, diluted HCl. Carrageenans were isolated by alcohol precipitation after an amylase treatment and a filtration of the extracts. The yields at 25 and 90 °C were 25 and 75% (w/w) of the dry alga, with molecular weights (M_w) corresponding to 194 000 and 245 000, respectively. The chemical structure was dominated by G4S-DA-(kappa-carrageenan or carrageenose 4-sulphate). A simple fractionation procedure for kappa-carrageenase hydrolysates, based on stirring in different enthanol/water mixtures, is introduced. NMR analysis showed that oligosaccharides with a repeating DA-G4S structure were the main constituents in the enzymic hydrolysates of the carrageenans from Hypnea musciformis. These oligosaccharides were solubilized in an ethanol concentration from 96 to 48% (v/v). In some enzyme resistant fractions D6S-G4S and DA2S-G4S sequences and D2S,6S unites were detected by ¹³C-NMR.Author for correspondence     

Purification and characterization of a new lectin from the red marine alga Hypnea musciformis

A lectin from the red marine alga Hypnea musciformis (HML) was purified by extraction with 20 mM PBS, precipitation with 70% saturated ammonium sulphate, ion-exchange DEAE-Cellulose chromatography and RP-HPLC. The 9.3 kDa polypeptide agglutinates erythrocytes from various sources and shows oligomerization tendencies under certain MALDI-TOF/MS conditions. Preliminary N-terminal sequencing and biological assays strongly suggest that the HML may belong to a new class of algae lectins.     

The effect of different concentrations of lead on inversion polymorphism in Drosophila subobscura

Drosophila subobscura is a wild Drosophila species that is spread over almost all of Europe. It possesses an uniquely rich inversion polymorphism on all five long chromosomes. This polymorphism is to a certain degree associated with the variation and dynamics of ecological factors in space and time. We analyzed the changes of inversion polymorphism components of Drosophila subobscura flies maintained on media with different concentrations of lead in laboratory conditions. The effects of lead on inversion polymorphism were observed by cytological analysis of gene arrangements on all of the five acrocentric chromosomes, as well as by cytological analysis of karyotypes on all of the four autosomes. The frequencies of particular gene arrangements on the four autosomes changed significantly in the samples maintained on medium not supplemented with lead. The frequencies of some gene arrangements on all of the five acrocentric chromosomes changed significantly in the flies maintained on media supplemented with lead. The length of exposure to different lead concentrations results in a significant change in the frequency of a few gene arrangements on two autosomes. However, the results show that different concentrations of lead, as well as the length of exposure, do not affect major parameters of inversion polymorphism. The results suggest that some gene arrangements could be linked with adaptive processes in evolving heavy metal resistance.     

Biomass and carrageenan yields of Hypnea musciformis in relation to selected environmental variables in the coastal waters of Ghana

Journal of Applied Phycology - Seaweed abundance and polysaccharide properties can vary spatially and temporarily, influenced by various environmental factors, although information on this is...     

不同基质栽培的药用鲍姆桑黄孔菌体外抗肿瘤活性的比较研究

比较3种不同栽培技术的鲍姆桑黄孔菌Sanghuangporus baumii——3年生栎树段木鲍姆桑黄孔菌、桑枝代料鲍姆桑黄孔菌和发酵鲍姆桑黄孔菌中多糖、总酚、黄酮及总三萜含量和体外抗肿瘤活性。分别采用硫酸-蒽酮、福林-酚、亚硝酸-硝酸铝和香草醛-冰醋酸法对4种主要成分进行含量测定。选用人大细胞肺癌细胞（H460）、人前列腺癌细胞（PC3）、人乳腺癌细胞（MDA-MB-231）、人肝癌细胞（SMMC-7721和BEL-7402）和人神经母细胞瘤细胞（SH-SY5Y）为待检细胞株,采用CCK-8检测细胞活力。结果显示,发酵鲍姆桑黄孔菌多糖含量最高（29.64%）,代料鲍姆桑黄孔菌和3年生段木鲍姆桑黄孔菌多糖含量均低于2.57%;2种代料鲍姆桑黄孔菌的总酚和黄酮含量较高,而发酵鲍姆桑黄孔菌最低（0.24%和0.68%）,尤其是代料鲍姆桑黄孔菌1号的总酚和黄酮含量高达4.02%和32.13%;三萜含量在4种栽培法的鲍姆桑黄孔菌中差异不明显。体外对6种肿瘤细胞增殖抑制能力最强的均为代料鲍姆桑黄孔菌,其IC₅₀值最低,其次是3年生段木鲍姆桑黄孔菌,发酵鲍姆桑黄孔菌在本研究设置的浓度范围内无明显的细胞毒性。结果表明：不同栽培方式鲍姆桑黄孔菌产生体外抗肿瘤活性不同,代料鲍姆桑黄孔菌最强,其次是3年生段木鲍姆桑黄孔菌,发酵鲍姆桑黄孔菌活性最差,这种活性差异与其有效成分含量高低直接相关;鲍姆桑黄孔菌发挥抗肿瘤活性的物质基础主要是总酚和黄酮。     

The coadaptation of female morphology and offspring size: a comparative analysis in crickets

Few studies of invertebrates have considered combinations of morphological and life history traits in the context of the evolution of reproductive strategies. Cricket species that exploit habitats harsh with respect to egg survival have evolved a long ovipositor, presumably because laying deep in the soil reduces egg mortality. Yet hatchling mortality increases with laying depth, and the ability of hatchlings to climb through the soil increases with egg size. Thus a conflict may exist between survival of the egg and that of the hatchling, inducing a positive covariation between egg size and ovipositor length across species evolving under contrasting selective habitats. We used the phylogenetic autocorrelation method and a path analysis to assess whether egg size coevolved with ovipositor length across 40 species of crickets, and whether egg size was affected by body size or ecological factors that influence egg mortality. Body size and ovipositor length were affected by taxonomic association, whereas common ancestry had no significant effect on egg size, diapausing strategy, and oviposition preference for soil types. The path model indicated that 29.11% of the variance in egg size was explained by independent evolution. As expected, ovipositor length was positively correlated with egg size, and species diapausing in the egg stage produced larger eggs than crickets diapausing in the nymphal stage or with no diapause. Ovipositor length and diapausing strategy were the first and second most important traits, respectively, in term of the proportion of variance in egg size explained by specific values. These results support the hypothesis that the ability of hatchlings to climb through the soil, and variation in diapause strategies, are general selective factors affecting the evolution of egg size in crickets. Phylogeny explained 51.01% of the variance in egg size. Egg size in a current cricket species, however, was not directly determined by egg size in its ancestor. Instead, it was strongly related to the phylogenetic values of body size and ovipositor length. Such indirect phylogenetic effects of body size and ovipositor length may have arisen because clades originating from ancestors with different ovipositor lengths experienced different selective pressures on egg size. Recelived: 13 October 1995 / Accepted: 30 September 1996     

The effect of different platelet-rich plasma concentrations on proliferation and differentiation of human periodontal ligament cells in vitro

OBJECTIVES: The use of platelets and platelet products has become increasingly popular clinically as a means of accelerating endosseous wound healing. It is likely that growth factors released by activated platelets at the site of injury play a role in periodontal regeneration by regulating cellular activity. The purpose of this study was to evaluate the biological effects of platelet-rich plasma (PRP) on human periodontal ligament cells (hPDLCs) in vitro. MATERIALS AND METHODS: Primary cultures of hPDLCs were obtained from healthy premolars. PRP was isolated by two-step centrifugation. Two main growth factors present in the thrombin-activated PRP (platelet-derived growth factor [PDGF-AB] and transforming growth factor-beta1 [TGF-beta1]) were evaluated using ELISA assay. Activated PRP or the combination of recombined human TGF-beta1 (rhTGF-beta1) and PDGF-AB (rhPDGF-AB) were added to hPDLCs in different concentrations to assess cell proliferation and osteogenic differentiation. RESULTS: PRP contained high levels of TGF-beta1 and PDGF-AB. Cell attachment, proliferation and ALP activity were enhanced by addition of PRP or rhTGF-beta1 and rhPDGF-AB combination to the cell cultures, while the stimulatory potency of PRP was much greater than the latter. These stimulatory effects presented in a dose-dependant manner, it seemed that PRP with 50~100 ng/ml TGF-beta1 was an ideal concentration. CONCLUSIONS: PRP can enhance hPDLC adhesion, proliferation and induce the differentiation of hPDLC into mineralized tissue formation cell; thereby contribute to the main processes of periodontal tissue regeneration. For economical and biological reasons, PRP has more clinical beneficial than analogous growth factors.     

The comparative analysis of the effect of oligopeptides and amino acids on the development of rat different tissue culture

The effect the synthetic tetra-, tripeptides and amino acids, composed them, in concentration 10(-12) M was investigated in organotypic tissue culture on the cell proliferation and apoptosis development in six tissue explants (ecto-, meso- and entodermal genesis) in rats at age 3 months old. The tetrapeptide demonstrated the greater tissue-specificity, as compared to the tripeptides and amino acids. Each tetrapeptide stimulated proliferation in only one corresponded tissue type. Less tissue-specificity was observed in the tripeptides, which stimulated proliferation and apoptosis in one (one case)--four tissues. The amino acids stimulated proliferation and apoptosis in three--five tissues. It can be suggested, that tissue-specificity in these biologically active substances is depended on the structure complexity. It is discussed, that the tetrapeptide tissue-specificity is related to the complementar interaction to the DNA site-specific blocks.     

The effect of lead ions on the energy metabolism of human erythrocytes in vitro

The aim of this work was to evaluate the influence of chronic exposure to lead ions on the parameters of energetic status of human erythrocytes in vitro. Umbilical cord erythrocytes were incubated with lead acetate at final lead ion concentrations ranging from 10 to 200 microg/dl. ATP, ADP, AMP, adenosine, GTP, GDP, GMP, guanosine, IMP, inosine, hypoxanthine, NAD and NADP concentrations in erythrocytes were determined using HPLC. Scanning electron micrographs of erythrocytes were taken. The mean concentrations of ATP, GTP, NAD and NADP, and mean values of adenylate energy charge (AEC) and GEC in cells incubated at the presence of lead ions were significantly lower after 20 h of incubation. Concentrations of purine degradation products (Ado, Guo, Ino) and Hyp were significantly higher. It is suggested that lead ions affect the energy metabolism of erythrocytes. Morphological changes in erythrocytes correspond to the increase of lead ions in the incubation mixture and to the decrease of ATP concentration in erythrocytes. A decrease in NAD and ATP concentration in erythrocytes could be a sensitive indicator of energy process disturbance, useful in monitoring in case of chronic lead exposure.     

Studies on lead and cadmium toxicity in Dianthus carthusianorum calamine ecotype cultivated in vitro

• Information on metallophytes during reclamation of land contaminated with heavy metals is sparse. We investigated the response of D. carthusianorum calamine ecotype to Pb and Cd stress. We focused on in vitro selection of tolerant plant material for direct use in chemically degraded areas.
• Shoot cultures were treated with various concentrations of Pb or Cd ions. Plantlet status was estimated as micropropagation efficiency, growth tolerance index (GTI) and through physiological analysis. Moreover, determination of plant Pb, Cd and other elements was performed.
• The application of Pb(NO₃)₂ resulted in stronger growth inhibition than application of CdCl₂. In the presence of Pb ions, a reduction was observed of both, the micropropagation coefficient to 1.1–1.8 and the GTI to 48%. In contrast, Cd ions had a positive influence on tested cultures, expressed as an increase of GTI up to 243% on medium enriched with 1.0 μm CdCl₂. Moreover, photosynthetic pigment content in shoots cultivated on media with CdCl₂ was higher than in control treatment. The adaptation to Cd was associated with decreased accumulation of phenols in the order: 0.0 μm  > 1.0 μm > 3.0 μm  > 5.5 μm CdCl₂. It seems that high tolerance to Cd is related to K uptake, which is involved in antioxidant defence.
• This work presents an innovative approach to the impact of Cd ions on plant growth and suggests a potential biological role of this metal in species from metalliferous areas.

     

不同钙浓度对宽叶雀稗幼苗的生长和抗性生理的影响

研究不同钙浓度对宽叶雀稗(Paspalum wettsteinii)幼苗生长和生理的影响, 对于揭示宽叶雀稗对不同钙浓度环境的适应机理至关重要。该研究采用盆栽砂培试验, 研究不同钙浓度(0、5、25、50、100和200 mmol·L^-1 CaCl₂)和不同处理时间(7、14、21和28天)对宽叶雀稗幼苗生长、渗透调节物质含量、抗氧化酶活性、叶绿素含量和光合参数的影响。结果表明, 随着CaCl₂浓度的增加和处理时间的延长, 宽叶雀稗幼苗株高等形态指标、生物量、渗透调节物质含量、抗氧化酶活性、叶绿素含量和光合参数呈先增后减的趋势, 低钙浓度(5-50 mmol·L^-1)环境下, 株高、叶长、叶宽、根长和生物量与对照(0 mmol·L^-1)相比均升高, 脯氨酸、可溶性蛋白和可溶性糖含量、过氧化物酶、过氧化氢酶和超氧化物歧化酶活性提高, 丙二醛含量和胞间CO₂浓度降低、叶绿素含量增加以及净光合速率、蒸腾速率和气孔导度增强; 高钙浓度(200 mmol·L^-1)环境下, 脯氨酸、可溶性蛋白和可溶性糖含量、过氧化物酶、过氧化氢酶和超氧化物歧化酶活性降低, 丙二醛含量和胞间CO₂浓度增加, 叶绿素含量减少以及净光合速率、蒸腾速率和气孔导度减弱。结合隶属函数分析, 低钙盐浓度(5-50 mmol·L^-1)处理对宽叶雀稗幼苗无抑制作用, 说明宽叶雀稗对低钙浓度具有一定的耐受性; 而在高钙浓度(200 mmol·L^-1)下, 宽叶雀稗幼苗通过提高自身有机渗透调节物质含量、增强酶活性、增加叶绿素含量以及增强光合作用等方式来快速调节植物生理代谢功能, 进而适应高钙浓度环境条件。     

Blue copper proteins: a comparative analysis of their molecular interaction properties

Blue copper proteins are type-I copper-containing redox proteins whose role is to shuttle electrons from an electron donor to an electron acceptor in bacteria and plants. A large amount of experimental data is available on blue copper proteins; however, their functional characterization is hindered by the complexity of redox processes in biological systems. We describe here the application of a semiquantitative method based on a comparative analysis of molecular interaction fields to gain insights into the recognition properties of blue copper proteins. Molecular electrostatic and hydrophobic potentials were computed and compared for a set of 33 experimentally-determined structures of proteins from seven blue copper subfamilies, and the results were quantified by means of similarity indices. The analysis provides a classification of the blue copper proteins and shows that (I) comparison of the molecular electrostatic potentials provides useful information complementary to that highlighted by sequence analysis; (2) similarities in recognition properties can be detected for proteins belonging to different subfamilies, such as amicyanins and pseudoazurins, that may be isofunctional proteins; (3) dissimilarities in interaction properties, consistent with experimentally different binding specificities, may be observed between proteins belonging to the same subfamily, such as cyanobacterial and eukaryotic plastocyanins; (4) proteins with low sequence identity, such as azurins and pseudoazurins, can have sufficient similarity to bind to similar electron donors and acceptors while having different binding specificity profiles.     

Accumulation of intermediates in a denitrifying process at different copper and high nitrate concentrations

NO2- accumulation occurred in an upflow anaerobic sludge blanket reactor at 500, 1000 and 1500 mg NO3- -N/l, but not after copper in the medium was increased to 28 g Cu2/l. N2O release was not avoided even with 56 g Cu2/l, at 1500 and 2500 mg NO3- -N/l and reached 251 mg N2O-N/ld. © Rapid Science Ltd. 1998