Precision and accuracy of the determination of the ionic product of 10750oxy-apatite

The formation of 10750oxy-apatite may be a major determinant of the concentration of dissolved orthophosphate in many natural hard fresh waters. Tests of this hypothesis require knowledge of the precision and accuracy of the calculated ionic product [Ca²⁺]⁵ [PO _inf4 ^sup3– ]³ [OH^–]. In routine analyses the largest contribution to overall error is probably that of pH: precision and bias better than 0.02 units are desirable. There is also a large uncertainty present in the possible bias of the third ionisation constant of ortho-phosphoric acid.     

Voltage clamping with single microelectrodes: Comparison of the discontinuous mode and continuous mode using the Axoclamp 2A amplifier

Summary The voltage clamp technique is a powerful method for studying the physiology of excitable membrane. This technique has made possible the determination of ionic responses generated by activation of either receptor-mediated or voltage-dependent processes. The development of the whole-cell, tight-seal voltage clamp method has allowed the analysis and examination of membrane physiology at the single cell level. The method allows the characterization of voltage-dependent ionic conductances both at the macroscopic (whole-cell) and at the microscopic (unitary conductance or single channel) level in cells less than 10 µm in diameter, a feat difficult to achieve with conventional fine-tipped micropipettes.In this paper, several methologies used for culturing neuronal and non-neuronal cells in the laboratory are described. A comparison between the two modes of voltage clamp using blunt-tipped patch-microelectrodes, the switching (discontinuous) and the non-switching (continuous) modes, of the Axoclamp-2A amplifier is made. Some results on membrane currents obtained from neuronal and non-neuronal cells using the single electrode whole-cell tight-seal voltage clamp is illustrated. The possible existence of two inactivating K⁺ currents, one dependent on Ca⁺⁺ the other is not, is discussed.     

Birth of 'human-specific' genes during primate evolution

Humans and other Anthropoids share very similar chromosome structure and genomic sequence as seen in the 98.5% homology at the DNA level between us and Great Apes. However, anatomical and behavioral traits distinguish Homo sapiens from his closest relatives. I review here several recent studies that address the issue by using different approaches: large-scale sequence comparison (first release) between human and chimpanzee, characterization of recent segmental duplications in the human genome and analysis of exemplary gene families. As a major breakthrough in the field, the heretical concept of human-specific genes has recently received some supporting data. In addition, specific chromosomal regions have been mapped that display all the features of gene nurseries and could have played a major role in gene innovation and speciation during primate evolution. A model is proposed that integrates all known molecular mechanisms that can create new genes in the human lineage.     

Metal interactions in algal toxicology: conventional versus in vivo tests

Different studies on the toxicity of the same metals to algae have often shown divergent and sometimes contradictory results. The inconsistency of these findings was often attributed to environmental factors such as the degree of chelation, complexation and precipitation of metals. Conventionally, toxicity tests of metal mixtures were conducted by adding metals to algae growing in synthetic media. In our study, we examined the in vivo toxic effects of metals by pretreating the algal cells with one metal, resuspendeding them in fresh medium, then exposing them to a second metal. The algal response showed marked differences between the conventional and the new approach. The conventional approach shows that the toxicity depends upon the complexes formed externally, whereas the in vivo approach shows that the toxicity probably depends upon the molecular transformation of the metals internally.     

Relationship between boron content and antioxidant compounds in Citrus leaves taken from fields with different water source

Boron contamination in the agricultural lands is an important problem for Western Turkey, which has rich boron deposits. This study was carried out in Nazilli regions upon orange (Citrus sinensis L. Osbeck) plants irrigated with relatively high boron laden channel waters (LCI) and with well waters (LWI) which contain lower amounts of boron. The leaves of the plants irrigated with channel water were found to contain twice the amount of boron compared with those irrigated with well waters. Boron content of leaves in both groups were approximately ten times the boron content in the soil on which they are grown. In the leaves of the plants irrigated with channel waters there were approximately 50% chlorophyll loss and higher chlorophyll a/b and caretonoid/chlorophyll ratios. In the excessive boron containing leaves was found higher soluble protein and carbohydrate contents, but lower determined free proline value. In plants that irrigated with high boron laden channel water significantly lower -tocopherol content and two fold higher ascorbate concentration were determined. The lower activities of catalase and glutathione reductase enzymes and higher total superoxide dismutase activity were measured in high boron content leaves. The retardation of growth due to boron toxicity can be attributed to the chlorophyll loss and inhibition of the carbohydratemetabolism. Boron at toxic level may cause the cell membrane lipids to be damaged by the free radicals by decreasing the -tocopherol levels. The increase in the ascorbate concentration may have a vital role in the protection of the inner cell structures against the boron toxicity.     

Assessment of possible sediment toxicity of contaminated sediments in Port Jackson, Sydney, Australia

A considerable amount of chemical data is available for surficial sediments in Port Jackson. Some of the highest concentrations of heavy metals, organochlorines and polycyclic aromatic hydrocarbons of any capital port occur in sediments mantling shallow tributaries and embayments close to central Sydney. However, these data have limited ability to predict adverse effects on living resources and in the absence of toxicological data, sediment quality guidelines (SQG) have been used to assess the possible adverse biological effects of sedimentary contaminants in this estuary. Several SQGs are currently available for both fresh and marine environments, but the scheme used in the current study is based on empirical analysis of matching chemical and biological data compiled by the National Oceanographic and Atmospheric Administration (NOAA) in the U.S. The NOAA SQG can be used to assess individual chemicals, or to estimate the probability of acute sediment toxicity by calculating `mean quotients' for a large range of contaminants. Although many individual chemicals in sediment exceed SQG over extensive parts of Port Jackson, `mean quotient' results suggest that only a small proportion (1%) of the harbour may be highly toxic (74% probability of toxicity). However, sediment in a considerably larger proportion of the port (almost 25%) is estimated to have a 49% probability of being toxic using the `mean quotient' approach. These results are indicative at best and contemporaneous chemical/biological/ecotoxicological studies are needed to verify the applicability of SQG developed in the U.S. for use outside North America and site specific studies of this type are still required to determine the toxicity of sediments in Port Jackson.     

Assessing the impact of sewage effluent on the ecosystem health of the Toronto Waterfront (Ashbridges Bay), Lake Ontario

The ecosystem health of the Toronto Waterfront (Ashbridges Bay), Lake Ontario which receives treated sewage effluent was investigated during 1987 and 1988 by means of a functional and structural battery of tests. The functional tests included in situ size-fractionated primary productivity, Algal Fractionation Bioassays (AFBs), unfiltered and filtered bioassays, and sediment assays with Daphnia magna and Hyalella azteca. The structural evaluation involved the biomonitoring of the components of the microbial loop, such as bacteria, autotrophic picoplankton, heterotrophic nanoflagellates, and protozoa. The experimental results reveal a diversity of physiological responses to the complex nutrient and contaminant regimes by the indigenous phytoplankton. There was no evidence of the impact of chlorination on the primary productivity of the Bay. The overall productivity was higher during the post-chlorination period than the pre-chlorination phase. The high rates of microplankton + netplankton productivity near the outfall have been attributed to the bioavailability of nutrients which, quite possibly, exert ameliorating effects on metal toxicity. In contrast, the low ultraplankton rates have been interpreted to be due to their well-known sensitivity to contaminants. The Effluent Receiving Water Bioassays (ERWB) with filtered and unfiltered experiments provided interesting insight and appear to be a potentially useful assessment tool. Generally, the unfiltered water compared to the filtered was toxic to the offshore test phytoplankton. This demonstrates a unique ecological adaptation to the prevailing in situ conditions by the Bay community which might be important from the restoration point of view. However, the offshore population was found to be sensitive to the particulate-bound toxicity as indicated by the unfiltered bioassays. Consequently, it is essential to probe the complexity of nutrient-contaminant interactions which ultimately appear to determine the toxicity and the resulting health of the biota. Furthermore, our experiments have shown that the particulate-matter is an important carrier of both nutrients and contaminants in Ashbridges Bay. The sediment bioassays for Station 419 indicated that sediments were toxic during both the pre- and post-chlorination phases. Both solid and liquid phase testing indicated toxicity of sediment to the acute Daphnia test. The Hyalella chronic assay showed good survival during the 4-wk period of the experiment, in contrast to the toxicity observed for phytoplankton and Daphnia. This may be due to large mounts of organic matter available in the Bay. The invertebrate bioassays confirmed the lack of impact of chlorination. Finally, the microbial loop seems to be a sensitive, rapid, and an early warning bioindicator of anthropogenic stress. The multi-trophic battery of structural and functional strategy adopted in our laboratory appear to be holistic and effective. The strategy has a considerable potential for developing eco-technology for a badly needed assessment and restoration of ecosystem health of the Great Lakes as well as other perturbed environments in the world.     

Corrections to the Chironomidae part of the Catalogue of Palaearctic Diptera

An extensive list of corrections to the Chironomidae part of the Catalogue of Palaearctic Diptera (ASHE and CRANSTON, 1990) is given.     

Ornithine decarboxylase prevents tumor necrosis factor alpha-induced apoptosis by decreasing intracellular reactive oxygen species

Ornithine decarboxylase (ODC) plays an essential role in various biological functions, including cell proliferation, differentiation and cell death. However, how it prevents the cell apoptotic mechanism is still unclear. Previous studies have demonstrated that decreasing the activity of ODC by difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, causes the accumulation of intracellular reactive oxygen species (ROS) and cell arrest, thus inducing cell death. These findings might indicate how ODC exerts anti-oxidative and anti-apoptotic effects. In our study, tumor necrosis factor alpha (TNF-) induced apoptosis in HL-60 and Jurkat T cells. The kinetic studies revealed that the TNF- -induced apoptotic process included intracellular ROS generation (as early as 1 h after treatment), the activation of caspase 8 (3 h), the cleavage of Bid (3 h) and the disruption of mitochondrial membrane potential ( _m) (6 h). Furthermore, ROS scavengers, such as glutathione (GSH) and catalase, maintained _m and prevented apoptosis upon treatment. Putrescine and overexpression of ODC had similar effects as ROS scavengers in decreasing intracellular ROS and preventing the disruption of _m and apoptosis. Inhibition of ODC by DFMO in HL-60 cells only could increase ROS generation, but did not disrupt _m or induce apoptosis. However, DFMO enhanced the accumulation of ROS, disruption of _m and apoptosis when cells were treated with TNF- . ODC overexpression avoided the decline of Bcl-2, prevented cytochrome c release from mitochondria and inhibited the activation of caspase 8, 9 and 3. Overexpression of Bcl-2 maintained _m and prevented apoptosis, but could not reduce ROS until four hours after TNF- treatment. According to these data, we suggest that TNF- induces apoptosis mainly by a ROS-dependent, mitochondria-mediated pathway. Furthermore, ODC prevents TNF- -induced apoptosis by decreasing intracellular ROS to avoid Bcl-2 decline, maintain _m, prevent cytochrome c release and deactivate the caspase cascade pathway.     

Genetically determined differences in the response of alpha1-antitrypsin levels in human serum to typhoid vaccine

Summary Normal individuals and those homozygous and heterozygous for the gene which determines deficiency in serum ₁-antitrypsin (₁-at) were given an intravenous injection of typhoid vaccine. Following this injection the trypsin-inhibiting capacity of serum and the concentration of ₁-at increases markedly in normal individuals. Heterozygotes for the deficiency gene have an elevation of ₁-at and trypsin-inhibiting capacity of their serum but reach only 50 percent of the levels reached by homozygotes for the common gene (normal individuals). Their values are however temporarily in the range of normal men not given typhoid vaccine. Homozygotes for the deficiency gene show only a small elevation of ₁-at under these conditions. These findings demonstrate that (1) the increase in trypsin-inhibiting capacity of serum after an injection of typhoid vaccine is largely due to the elevation of the ₁-at concentration, and (2) the ₁-at deficiency gene inhibits the quantitative response of the ₁-at to such a stimulus.Supported in part by USPHS grant HE-06285 from the National Heart Institute.Some of these studies were carried out in the General Clinical Research Center Ward, U. C., FR-79, supported by the Division of Research Facilities and Resources, U.S. Public Health Service.     

Improved rate of callus induction from rice anther culture following microscopic staging of microspores in iron alum-haematoxylin

Summary High frequencies of callusing were obtained in indica rice from the microspores which were staged in acetic acid iron alum-haematoxylin stain prior to culture on G5 medium. Two local varieties, Khonorullo and Namyi, and two advance pre-release cultures, PK 1-1-3 and PK 12-22, were used in this investigation. All the cultures exhibited a wide adaptation to varying medium; however, the frequency of callusing was highest (45.5%) in PK 1-1-3 followed by PK 12-22 (32.4%) and Khonorullo (31.6%). Cold shock (10 °C) for 11 days enhanced the frequency of callusing by 200% in Khonorullo.     

Chemical characteristics of pond waters in the Labyrinth of southern Victoria Land,Antarctica

A large number of fresh water and saline ponds are located in the Labyrinth (77° 33 S, 160° 50 E) of the upper Wright Valley in the Dry Valleys region of southern Victoria Land, Antarctica. It is situated near the terminus of the Wright Upper Glacier between 800–1000 m above sea level. From a limnological point of view, the most interesting problems concerning these saline ponds are the origin of their salts and their evolutional history.Chloride ion contents vary remarkably among the ponds ranging from 0.0049 to 52.4 g kg^–1. Surprisingly, more than a half of the ponds are saline with the highest chloride ion content being 2.7 times greater than that of seawater. The D and ¹⁸O values of the pond waters indicate a snow and/or glacial meltwaters origin, and that the ponds underwent subsequent alteration due to evaporation or freezing. The composition of chemical components reveal no evidence of trapped seawater. Thus the salt concentrations in the Labyrinth pond waters must be explained principally by the accumulation of atmospheric salts and subsequent repeated cycles of evaporation and freezing of the pond waters over considerable time periods.     

Temporal variation of phytophilous Chironomidae over a 11-year period in a shallow Neotropical lake in southern Brazil

Small and shallow lakes make up most of the lake-covered area in the world, and immature Chironomidae generally constitute the dominant group among invertebrates in these systems, exhibiting a wide distribution and tolerance to various levels of water quality. The changes in trophic features associated with the dominance of different major primary producers (macrophyte and phytoplankton) are reflected in the functioning of these ecosystems and likely in their phytophilous Chironomidae species richness and abundance. These were thus studied in a Brazilian shallow lake over a 11-year period characterized by both clear and turbid water conditions in relation to water quality and dominant macrophytes. The results revealed differences in Chironomidae composition but not in richness associated with clear and turbid waters. The Goeldichironomus genera predominated during all years and for different macrophyte types (freefloating, floating leaf, and submerged). We attribute the variability of Chironomidae assemblages to changes in macrophyte dominance and water trophic status, such as the concentration of nutrients and chlorophyll-a, during the 11-year period. Submerged macrophytes were associated with clear waters and harbored the highest diversity, which emphasized the importance of these environmental conditions for preserving the biodiversity of Chironomidae in shallow lakes.     

Immunocytochemical study by two photon fluorescence microscopy of the distribution of GABA<Subscript>A</Subscript> receptor subunits in rat cerebellar granule cells in culture

Summary. An immunocytochemical investigation of the expression of ₁, ₆, _2/3, ₂ and subunits was performed on rat cerebellum granule cells in culture by the two photon microscopy technique.The first four subunits appear to be expressed abundantly in these cells, whereas the one seems to be expressed at a lower level. Another major difference in the distribution of these subunits is that whereas ₆, _2/3 and ₂ appear only on plasma membranes ₁ and are present mainly in the cell bodies cytoplasm. Still another difference was found in that the presence of ₂ on neurites is polarized, preferentially labelling neurites with the appearance of dendrites. The subunits ₆ and _2/3 appear to label all types of neurites, with _2/3 being by far the most heavily expressed subunit type. A final distinct characteristic is that ₆ and, even more, ₂ appear to accumulate in the cytoplasmic domains immediately below the cone of emergence of neurites. This suggests a conspicuous transport of such subunits from the site of synthesis in the cell body to the site of final expression in the neurites (dendrites and axon terminals).     

Interpretations of gradients in δ13C value in thick photosynthetic tissues of plants with Crassulacean acid metabolism

In Ceropegia dichotoma, Crassula argentea, Esheveria colorata, Kalanchoë beharensis, Opuntia ficus-indica, Sansveria stuckyi and Opuntia inermis the carbon-isotope ratio ( ¹³C) of tissues close to the epidermis is 2–4.3 more negative than those in the centre of the leaf or cladode. The greatest change in ¹³C value occurs between the epidermal layer and the layer of mesophyll tissue immediately underneath. Analysis of major metabolic and structural components in successive layers of Crassula argentea grown under controlled environmental conditions conducive to Crassulacean acid metabolism confirmed that much of the variation in ¹³C values of bulk carbon is caused by differences in chemical composition. Thus the steep gradient in ¹³C value at the epidermis reflects, in part, the contribution of more-negative ¹³C values of lipids in these tissues. Moreover, during nocturnal CO₂ fixation the amount of malic acid synthesised decreases with depth and the ¹³C value of the methanol-soluble fraction is less negative with distance away from the upper epidermis. These results are consistent with diffusion limitation to CO₂ uptake in these thick leaf tissues, which also contributes to the observed gradients in ¹³C value.Abbreviation CAM Crassulacean acid metabolism - FW fresh weight - PEPCase phospoenolpyruvate carboxylase - Rubisco ribulose-1,5-bisphosphate carboxylase-oxygenase - ¹³C carbon-isotope ratio This work was supported by grants from the North Carolina Biotechnology Centre to Duke University, National Science Foundation (NSF) grant DCB90-06830, Department of Energy, Division of Energy Biosciences grant DE-FG05-89 ER 14005, and NSF grant BSR 87-06429 to Duke University Phytotron.     

Re-assessment of Wundsch's (1940) ‘H2S-Oscillatoria-Lake’ type using the eutrophic Lake Scharmützel (Brandenburg, NE Germany) as an example

Thienemann's 1922 biological lake classification indicates the increase in the trophic level of lakes. It is based on the sucession of the disappearance of certain benthic indicator organisms in relation to the gradient of a decreasing O₂concentration in the deep water, i.e. from O₂-sensitive Chironomidae spp. (non-biting midges) to the less sensitive Chaoboridae (phantom midges) larvae. As early as the mid 1930s, several lakes in Brandenburg, e.g. Lake Scharmützel, belonged to the last category of this classification. They were O₂-deficient in deep water during summer and lacked the Chironomidae larvae. Simultaneously, filamentous cyanobacteria (Oscillatoriales) appeared. This lead to the replacement of Thienemann's indicators, i.e. O₂ by H₂S and Chironomidae by Oscillatoriales, and to Wundsch's 1940 new H₂S-Oscillatoria-Lake type. Since H₂S and Oscillatoriales were not clearly identified as symptoms of eutrophication, it is not justified to use them to characterise a separate lake type. However, these indicators are of ecological importance, since the regional and common creeping increase in SO₄ ^2- concentration favoures the current high H₂S formation. The successive deterioration in O₂ conditions, the increase in the extent of H₂S formation related to the decrease in species diversity clearly indicates that Lake Scharmützel reached a qualitatively new state of eutrophication within 60 years.     

Molecular dynamics simulations of ribonuclease T1

Molecular dynamics simulations in vacuum and with a water sphere around the active site were performed on the 2GMP-RNase T1 complex. The presence of water led to the maintenance of the 2-GMP-RNase T1 interactions as compared to the X-ray structure, including the hydrogen bonds implicated in the enzyme-inhibitor recognition process. The sidechain of His92 in the molecular dynamics water simulation, however, hydrogen bonds directly to the phosphate of 2GMP in contrast to the X-ray structure but in support of the role of that residue in the enzyme's catalytic mechanism. Fluctuations of activesite residues are not strongly influenced by water, possibly owing to the exclusion of water by the bound 2GMP, which did show an increase in mobility. Analysis of the 2GMP-RNase T1 interactions versus time reveal an equilibrium fluctuation in the presence of water, leading to a less favorable 2GMP-RNase T1 interaction energy, suggesting a possible relationship between picosecond fluctuations and inhibitor dissociation occurring in the millisecond time domain.Abbreviations RNase T1 Ribonuclease T1 (EC.3.1.27.3) - 2GMP Guanosine-2-monophosphate - SBS Stochastic Bondary Simulation - VS Vacuum Simulation - MD Molecular Dynamics     

Gene duplication in salmonid fishes: Evidence for duplicated but catalytically equivalent A4 lactate dehydrogenases

Skeletal muscle tissues from many species of salmonid fish are known to exhibit a set of three to five isozymes for A₄-type lactate dehydrogenase (l-lactate: NAD oxidoreductase, E.C. 1.1.1.27), but the genetic basis for this isozyme system has not previously been assessed. This isozyme system was purified to homogeneity from salmon (Oncorhynchus tshawytscha) and shown to be composed of two polypeptides, A and A, in binomial tetrameric combinations. Amino acid analysis revealed that A and A are closely related but genetically distinct proteins, and thus are coded for by recently duplicated structural loci. Catalytic studies on the purified intact salmon isozyme system and on the isozymically pure A₄ and A₄ homotetramers from brown trout (Salmo trutta) revealed no significant differences in catalytic properties among these enzymes, suggesting equivalent catalytic function of A and A. These results, in combination with studies on polypeptide and isozyme ratios, suggest that one of the duplicated loci in salmon may be drifting toward a nonfunctional state by accumulation of mutations in regulatory DNA rather than in the structural gene itself.This work was supported in part by research grants from the University Grants Committee, the University of Otago Research Committee, and the Medical Research Council of New Zealand. Part of this work was performed while one of us (S. T. L.) was supported by predoctoral fellowships from the Medical Research Council of New Zealand and from the Lee Foundation.     

Is haemoglobin Gα Philadelphia linked to α-thalassaemia?

Summary The question, Is Hb G Philadelphia linked to -thalassaemia? was first posed because the abnormal haemoglobin is found in heterozygotes at a concentration greater than 25%, the proportion predicted from a 4 -chain gene model. Globin chain biosynthesis was studied in a West Indian family in which one parent had ⁺ thalassaemia and the other was heterozygous for the G Philadelphia chain gene. The former had a globin chain production ratio / well above 1, while the latter had a ratio significantly less than 1. One child of the marriage had inherited the ⁺ thallassaemia from one parent and the G Philadelphia chain gene from the other and showed the typical picture of /-thalassaemia (/ ratio slightly above normal). It is explained in the discussion that the evidence favours a close linkage of 2 -chain genes.