Ecological determinants of methylmercury bioaccumulation in benthic invertebrates of polar desert lakes

We investigated concentrations of monomethylmercury (MMHg) at the base of benthic food webs in six lakes from polar desert (biologically poor and low annual precipitation) on Cornwallis Island (Nunavut, Canada, ~75°N latitude). Anthropogenic mercury emissions reach the Arctic by long-range atmospheric transport, and information is lacking on processes controlling MMHg entry into these simple lake food webs, despite their importance in determining transfer to lake-dwelling Arctic char. We examined the influences of diet (using carbon and nitrogen stable isotopes), water depth, and taxonomic composition on MMHg bioaccumulation in benthic invertebrates (Chironomidae and Trichoptera). We also estimated MMHg biomagnification between benthic algae and invertebrates. Similar MMHg concentrations of chironomid larvae in nearshore and offshore zones suggest that benthic MMHg exposure was homogeneous within the lakes. Chironomid δ¹³C values were also similar in both depth zones, suggesting that diet items with highly negative δ¹³C, specifically methanogenic bacteria and planktonic organic matter, were not important food (and therefore mercury) sources for profundal larvae. MMHg concentrations were significantly different among two subfamilies of chironomids (Diamesinae, Chironominae) and Trichoptera. Higher MMHg concentrations in Diamesinae were likely related to predation on other chironomids. We found high MMHg biomagnification between benthic algae and chironomid larvae compared with literature estimates for aquatic ecosystems at lower latitudes; thus, benthic processes may affect the sensitivity of polar desert lakes to mercury. Information on benthic MMHg exposure is important for evaluating and tracking impacts of atmospheric mercury deposition and environmental change in this remote High Arctic environment.     

Immunophenotypic Characterization of CD45RO+ and CD45RA+ T Cell Subsets in Peripheral Blood of Peripheral T Cell Lymphoma Patients

To study the distribution profile of CD45RO⁺ and CD45RA⁺ T cells in the peripheral blood of peripheral T cell lymphoma (PTCL) patients and its clinical significance. 27 patients with PTCL were enrolled in this study, together with 30 healthy individuals as the control group. Flow cytometry analysis was employed to examinate the differences in the distribution of CD45RO⁺ and CD45RA⁺ T cells in peripheral blood between two groups. In PTCL patient’s lymphnode tissues, the T cell population displayed diverse antigenic expression, with CD4⁺ T cells as the major subset. No B cell-related antigen was expressed. The percentage of CD4⁺/CD8⁺ and CD4⁺CD45RO⁺ T cells in patients’ peripheral blood were significantly lower than that in the control samples, while the percentage of CD4⁺CD45RA⁺, CD8⁺CD45RA⁺, and CD8⁺CD45RO⁺ T cells in patients’ peripheral blood were significantly higher than that in the control samples. The percentage of CD4⁺/CD8⁺, CD4⁺CD45RO⁺ cells in stage I/II PTCL patients’ peripheral blood were significantly higher than that in the samples from patients with stage III/IV PTCL. The percentage of CD4⁺CD45RA⁺, CD8⁺CD45RA⁺, and CD8⁺CD45RO⁺ T cells were notably lower than that in the samples from III/IV period PTCL patients. Both CD45RO⁺ and CD45RA⁺ T cells play important roles in the process of PTCL. The immunophenotypic profile from this study will help to develop the differential diagnosis and treatment of PTCL patients in the future, and improve the accuracy rate of diagnosis and to ameliorate the prognosis.     

RNA and Macronuclear transcription in the ciliate Stylonychia mytilus

Nuclear and cytoplasmic RNA of Stylonychia mytilus were analyzed on denaturing polyacrylamide gels. The molecular weight of rRNA precursor molecules is within a range of 2.1×10⁶ daltons. A comparison between the electrophoretic pattern of nuclear non-ribosomal RNA and cytoplasmic mRNA indicates that a considerable amount of nuclear RNA sequences is of higher molecular weight than cytoplasmic RNA sequences. The molecular weight distribution of cytoplasmic RNA supports the assumption that also in Stylonychia an average sized mRNA molecule contains 1,200–1,500 nucleotides according to a molecular weight of 4×10⁵ to 5×10⁵ daltons. The size of the polyadenylic acid fragment of poly-A⁺ RNA molecules is about 120 nucleotides. The total mass of cytoplasmic RNA is around 7.5/10¹⁰ g/cell, corresponding to 1.2×10⁷ average sized mRNA molecules per cell. RNA excess hybridization experiments show that 60% of the DNA sequences are transcribed into nuclear RNA and that the cytoplasmic mRNA sequences are homologous to about 40% of macronuclear DNA sequences. There is no indication of different frequency classes within the mRNA. The number of different mRNA species in a Stylonychia cell is 1.2–1.5×10⁴. On the average each of them is present about 1,000 times in every cell.     

The modern distribution of chironomid sub-fossils (Insecta: Diptera) in Costa Rica and the development of a regional chironomid-based temperature inference model

Chironomids have been shown to provide robust reconstructions of past temperature change and variability. This is the first study to assess the contemporaneous relationship between the distribution of sub-fossil chironomids and limnological and climatic parameters in Central America. Here, we describe the distribution of chironomids in a suite of 51 lakes in Costa Rica. We identify environmental variables that account for a statistically significant amount of variance in midge distribution, and develop a quantitative chironomid-based inference model for mean annual air temperature (MAT). Psectrocladius, which is documented for the first time in Costa Rica, dominate high-elevation lakes characterized by low MAT and relatively dilute water. Canonical correspondence analysis (CCA) revealed that MAT and conductivity account for large, statistically significant amounts of variance in the distribution of chironomids. A chironomid-based inference model for MAT, developed using a partial least squares approach, provided robust performance statistics with a high coefficient of determination and a relatively low root-mean square error. Application of the chironomid-based inference model for MAT to chironomid stratigraphies spanning the Holocene, together with the ecological information provided by this study, will enable us to address many outstanding questions relating to long-term climate and environmental change in the region.     

The effect of 2-thiouracil on RNA synthesis in pollen tubesof Nicotiana alata

The level of RNA in pollen is approximately 20 mg g^-1 and remains constant during 6 h pollen germinationin vitro also in the presence of 2-thiouracil which stimulates pollen tube elongation. The synthesis of RNA in pollen tubes was investigated according to the incorporation of the label from uracil-2-¹⁴C, 2-thiouracil-2-¹⁴C, orotic acid-5-³H, fructose-U-¹⁴C and from³²PO₄ ^3- into RNA fractions separated by methylated albumine kieselguhr chromatography. The distribution of radioactivity on elution profiles was different according to the radioactivity source, however it was not changed by the presence of 2-thiouracil in cultivation medium. 2-Thiouracil incorporates into pollen tube RNA at about 50% the rate of uracil. It inhibited the incorporation of orotic acid, of fructose and of phosphate into all RNA fractions. It is suggested that the analogue inhibits the enzymes involved in RNA synthesis essentially as 2-thiouridine-5’-phosphate.     

High Fungal Diversity and Abundance Recovered in the Deep-Sea Sediments of the Pacific Ocean

Knowledge about the presence and ecological significance of bacteria and archaea in the deep-sea environments has been well recognized, but the eukaryotic microorganisms, such as fungi, have rarely been reported. The present study investigated the composition and abundance of fungal community in the deep-sea sediments of the Pacific Ocean. In this study, a total of 1,947 internal transcribed spacer (ITS) regions of fungal rRNA gene clones were recovered from five sediment samples at the Pacific Ocean (water depths ranging from 5,017 to 6,986 m) using three different PCR primer sets. There were 16, 17, and 15 different operational taxonomic units (OTUs) identified from fungal-universal, Ascomycota-, and Basidiomycota-specific clone libraries, respectively. Majority of the recovered sequences belonged to diverse phylotypes of Ascomycota (25 phylotypes) and Basidiomycota (18 phylotypes). The multiple primer approach totally recovered 27 phylotypes which showed low similarities (≤97 %) with available fungal sequences in the GenBank, suggesting possible new fungal taxa occurring in the deep-sea environments or belonging to taxa not represented in the GenBank. Our results also recovered high fungal LSU rRNA gene copy numbers (3.52?×?10⁶ to 5.23?×?10⁷copies/g wet sediment) from the Pacific Ocean sediment samples, suggesting that the fungi might be involved in important ecological functions in the deep-sea environments.     

Efficiency of modifications in the design and use of submerged funnel traps for sampling Chironomidae (Diptera)

The effects of three modifications in design and use of submerged funnel traps on capture efficiencies for emerging Chironomidae were examined in a recently-created reservoir in northern Manitoba, Canada. The experiments showed that: (1) Sample counts were higher in traps suspended just below the surface of the water than in traps suspended 1.5 m deeper, just above the lake bottom, probably owing to difficulties in handling the deep traps. (2) Sample counts were lower in traps with small cones inserted into the neck of the collecting bottle than in traps without these cones, although the differences were not significant. Bottles fitted with mini-cones usually retained higher numbers of other invertebrates which destroyed much of the chironomid catch. (3) Sample counts were proportionally higher in traps with a basal area of 0.283 m² compared to traps of 0.1 m² area.     

Regulation of sodium in the shore crabCarcinus maenas,adapted to environments of constant and changing salinities

The activity of Na-K-ATPase was determined in the posterior gills of the shore crabCarcinus maenas during a period following transfer from 35 to 10 ‰ salinity and vice versa at 15 °C. After transfer from high to low salinity, Na-K-ATPase activity increased from 3.2 to 7.0 μmoles P_i mg protein^?1 h^?1 within a period of 2 to 3 weeks. Transfer of crabs from low to high salinity resulted in reduction of activity from 7.4 to 4.5 μmoles P_i mg protein^?1 h^?1 within about the same period. The relatively slow response following salinity change indicates that the amounts of Na-K-ATPase in the gills may play a role in hyperionic Na regulation in relatively constant brackish-water environments. Instant responses to salinity result from activation and inhibition of Na-K-ATPase activity by Na. Gill Na-K-ATPase is activated by the Na concentration of the incubation medium to attain a steep maximum at about 75 mM Na, which corresponds to the lowest environmental Na levels tolerated byC. maenas equivalent to a salinity of ca 6 ‰. Activity greatly decreased towards higher Na levels, equivalent to the salinity of normal sea water, at which hyperregulation no longer occurs. Selective addition of either Na or Cl to brackish water of 9 ‰ S resulted in effective hyperregulation of the non-increased ion, and passive distribution between medium and blood of the increased ion. These data indicate that under appropriate conditions the normally coupled transport of Na and Cl may be uncoupled and take place independently of each other.     

De novo expression of fetal ED-A+ fibronectin and B+ tenascin-C splicing variants in human cardiac allografts: potential impact for targeted therapy of rejection

Management of acute and especially chronic rejection after human cardiac transplantation is still challenging. Chronic rejection, represented by allograft vasculopathy (CAV) and cardiac interstitial fibrosis (CIF) is known to cause severe long-term complications. Rejection associated tissue-remodelling entails the reoccurrence of fetal variants of Fibronectin (Fn) and Tenascin-C (Tn-C), which are virtually absent in adult human organs. In a rat model, an extensive re-expression could be demonstrated for ED-A⁺ Fn with spatial association to CAV and CIF. Thus, it is of great interest to investigate the cardiac tissue expression and distribution in human samples. From 48 heart transplanted patients, 64 tissue specimens derived from right ventricular biopsies were available. Histopathological analysis was performed according to the International Society for Heart and Lung Transplantation (ISHLT) guidelines for the detection of acute rejection. By immunohistochemistry, protein expression of ED-A⁺ Fn, B⁺ Tn-C, alpha-smooth muscle actin, CD31 and CD45 was assessed and analysed semiquantitatively. Co-localisation studies were performed by means of immunofluorescence double labelling. Histopathological analysis of the 64 samples revealed different ISHLT grades (0R in 36 cases, 1R in 20 cases and 2R in 8 cases). There was a distinct and quantitatively relevant re-occurrence of ED-A⁺ Fn and B⁺ Tn-C in most samples. Semi-quantitative evaluation did not show any correlation to the acute rejection grade for all markers. Interestingly, significant correlations to the extent of inflammation could be shown for ED-A⁺ Fn (r = 0.442, p = 0.000) and B⁺ Tn-C (r = 0.408, p = 0.001) as well as between both proteins (r = 0.663, p = 0.000). A spatial association of ED-A⁺ Fn and B⁺ Tn-C to CAV and CIF could be demonstrated. A relevant re-occurrence of ED-A⁺ Fn and B⁺ Tn-C following human heart transplantation could be demonstrated with spatial association to signs of rejection and a significant correlation to tissue inflammation. These data might contribute to the identification of novel biomarkers reflecting the rejection process and to the development of promising strategies to image, prevent or treat cardiac rejection.     

On the existence of two GABA pools associated with newly synthesized GABA and with newly taken up GABA in nerve terminals

[¹⁴C]Glutamic acid and [³H]GABA were injected into the lateral ventricle of mouse and then [¹⁴C]GABA and [³H]GABA in synaptosomes isolated from the animals were analysed. The [¹⁴C]GABA was interpreted to be newly synthesized GABA from [¹⁴C]glutamic acid while the [³H]GABA to be newly taken up GABA. We have obtained the following results: (1) when the animals were pretreated with aminooxyacetic acid and thus the GABA content in synaptosomes increased to about 2 times of the control level, only the [³H]GABA was enhanced to 3 times of the control level without any change of [¹⁴C]GABA, (2) the release of [¹⁴C]GABA from synaptosomes by high K⁺ depolarization was 1.5 times greater than that of [³H]GABA, (3) the releases of both [¹⁴C]GABA and [³H]GABA were increased in the presence of cold GABA,l-2,4-diaminobutyric acid or γ-amino-β-hydroxybutyric acid, but only slightly increased in the presence of β-alanine. These results would suggest that newly synthesized GABA and newly taken up GABA localized individually in different pools, which might localize either in different nerve terminals or separately in the same nerve terminal.     

Pistillate flower development and pollen tube growth mode during the delayed fertilization stage in Corylus heterophylla Fisch

Unlike most angiosperms, in which fertilization occurs within several days after pollination, fertilization in hazel (Corylus Spp.) is delayed by two to three and a half months. However, the female inflorescences or young fruits are too hard or lignified to be dissected according to regular paraffin sectioning technique. So, what the nature of development during the extended progamic phases of hazel remains unknown. The female inflorescence development and pollen tube growth mode during the delayed fertilization stage in hazel were investigated by improved paraffin sectioning and aniline blue staining of pollen tubes. The results showed ovaries and ovules of hazel were invisible at the time of blooming. Early ovary and ovule primordium began to form from 15 to 20 days after blooming, respectively. Integument and mature embryo sacs differentiated from the nucellus on 40th and 55th day after blooming, respectively. Pollen tubes were retarded in the bottom of the style or the pollen tube cavity (PTC, a specifical lignified cavity structure at the bottom of style for pollen tube to rest during progamic phase) for about 26 days. Then, the pollen tubes were observed to leave the PTC and began to enter the ovary. After that, a single pollen tube passed through the vicinity of the micropyle. Finally, pollen tubes turned a corner and penetrated the embryo sac through the tissue of the chalaza instead of micropyle on 52 and 55 days after blooming, respectively. The results of more in-depth information will be beneficial to better understanding of the delayed fertilization process in hazel.     

On the nature of RNA synthesized in pollen tubes ofNicotiana alata

The RNA formed in pollen tubes during 4 hours of growthin vitro was resolved by chromatography on methylated albumine on kieselguhr (MAK) into three principal fractions. Acoording to the labelling from uracil-¹⁴C about 11% was eluted with tRNA and 5 S RNA (low molecular weight RNA), 76% just after rRNA (D-RNA) and nearly 14% was recovered from the column by SDS at 35 °C (TB-RNA). In the presence of actinomycin D at concentration of 30 μg ml^-1 the synthesis of the three classes of RNA was inhibited by 71%, 97% and 70% respectively. On sucrose density gradient the radioactive low molecular weight RNA sedimented at 4 S-5 S which suggests that one or both of these RNA species are synthesized in pollen tubes. The D-RNA eluted from the MAK column is polydisperse in size exhibiting a wide range of sedimentation values up to about 35 S with a large peak at 9 S-10 S and two smaller peaks at 14 S-15 S and at about 23 S. The rapid labelling and the polydisperse rather low molecular weight character suggest that the D-RNA is a heterogeneous population of mRNA. The sedimentation profile of TB-RNA was similar to that of D-RNA. The RNA synthesized in the presence of³²PBO3-₄ or uracil-¹⁴C exhibited no radioactivity peaks corresponding to sedimentation peaks of rRNA.     

Effect of long-term shock-avoidance training on certain biochemical constituents of rat brain

A few constituents of the brain of rats subjected to long-term shock avoidance training in the jump-box were investigated. The effects of training and that of the concomitant stress on these constituents were differentially examined. Brain DNA increased in both the trained rats and those subjected to stress. The total RNA showed a specific decrease in the trained rats. The total protein content remained unaffected, while the content of soluble protein showed a greater increase in the trained rats compared to that in animals subjected to stress. The Na⁺, K⁺-ATPase activity in the brain decreased in the trained rats, whereas the glial carbonic anhydrase activity increased in the stressed as well as the trained rats. No change was found in the activities of Mg⁺⁺-ATPase and pseudocholinesterase in the experimental groups.     

Location of the genes coding for 18S and 28S ribosomal RNA in the human genome

³H-rRNA obtained from Xenopus laevis tissue cultured cells, or a ³H-cRNA made from Xenopus ribosomal DNA, was used for heterologous in situ hybridisation with human lymphocyte metaphase chromosomes. Prior to hybridisation, chromosome spreads were stained with Quinacrine and selected cells showing good Q-banding photographed; the same cells were then rephotographed after autoradiography and pairs of photographs for each cell were used to make dual karyotypes. The chromosomes within each karyotype were divided into equal sized segments (approx. 0.7 μ), with a fixed number of segments for each chromosome type. The distribution of silver grains between segments showed that the ³H-RNAs hybridised specifically to the nucleolar organising regions of the D and G group chromosomes with no other sites of localised labelling in the complement. Control experiments showed no localisation, with insignificant labelling, when metaphase spreads were incubated in a mixture containing Xenopus ³H-rRNA and competing cold human (HeLa) rRNA. Filter hybridisation experiments on isolated human DNA showed that the Xenopus derived ³H-RNAs hybridised to a fraction of human DNA which was on the heavy side of the main DNA peak and that these RNAs were competed out in the presence of excess cold human rRNA, confirming the specificity of the heterologous hybridisation. In situ hybridisation experiments were also carried out on cells from individuals with one chromosome pair showing heteromorphism for either a very long stalk (nucleolar constriction) subtending a satellite, or a large satellite. It was shown that the chromosome with the large stalk hybridised four times as much ³H-rRNA as its homologue, whereas differences in the sizes of the subtended satellites did not materially affect hybridisation levels indicating that rDNA is located in the stalks and not the satellites. The amount of ³H-rRNA hybridised differs between chromosomes and individuals; these differences are heritable and rDNA can be detected by in situ hybridisation in all three chromosomes number 21 in cells from Down's patients and in translocated chromosomes conta.ining a nucleolar constriction. Different D and G group chromosomes which hybridised equal amounts of ³H-rRNA participated in rosette associations at metaphase in a random fashion in some individuals and in a non-random fashion in others. In all individuals studied chromosomes with large amounts of rDNA were not found to be preferentially involved in association. It was therefore concluded that the probability of a chromosome being involved in the formation of a common nucleolus is not a simple function of its rDNA content and other possible factors are considered.     

Comparison of the sensitivity of the serological latex and ELISA tests

A sensitivity of the serological latex and ELISA tests were compared in carnation mottle virus diagnosis. For the latex test carnation mottle virus (CaMV) antiserum was sensibilized with latex suspension for RF-test. Sensibilized antiserum was used in 1: 200 dilution, as compared with fresh antiserum. For ELISA the γ-globuline fraction of antiserum was conjugated with alkaline phosphatase. The optimal dilution in both, CaMV fraction of antisera for coating of plates and γ-globuline-enzyme conjugate were in the ratio of 1: 500, 2 μg of antibodies in 1 ml. The dilution end point of carnation mottle virus in sap from carnation leaves was 1.6 × 10^?4 to 1.25 × l0^?5 and 1 × 10^?4 to 1.25 × l0^?5, when serological latex and ELISA tests were used. As indicated, ELISA as compared to the latex test was found to be more sensitive for carnation mottle diagnosis. As the latex test is considered to be simpler and cheaper, and in addition, showing the same assurance as the biological test onChenopodium amaranticolor, the latex test is recommended for carnation mottle virus detection.     

Ecological survey of the night monkey,Aotus trivirgatus,in Formosa Province,Argentina

Transect surveys were carried out in northern Argentina during October and November 1977 in order to determine the distribution and abundance ofAotus trivirgatus. The monkeys were seen in pairs with one to two recent young and occurred at a density of approximately six family groups/km².Aotus was only found in relatively moist, riparian, low forests. Some life history traits, such as diurnal activity and the lack of tree-hole use, are distinctive compared to more northern populations.