Human alfa-fetoprotein: isolation and production of monoclonal antibodies.

Alfa-fetoprotein from human cord serum was purified in a single step by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B with a final recovery of alfa-fetoprotein of about 90% and a purification factor of 900. The purified preparation was homogeneous on SDS-PAGE and native PAGE running with a relative molecular weight of 72,000. Monoclonal antibodies against this purified preparation were raised by hybridoma technology using Sp2/0 myeloma cells as a fusion partner. 50% of culture wells exhibited hybrid growth and 7% of these wells contained anti-AFP secreting hybrids. Positive hybrid cells were cloned twice by the limiting dilution method and 8 clones were obtained that secreted monoclonal antibodies. Five of these cell lines (3F6H10, 3F6H4, 3F6H1, 3F6G5 and 3F6G10) were selected at random for purification and characterization purposes. All 5 cell lines secreted monoclonal antibodies of IgG1 subclass which were purified by affinity chromatography on Protein A- Sepharose CL-4B column with a final recovery of 80% and a purification factor of about 13. The purified preparations were homogeneous on SDS-PAGE, native PAGE and IEF. The monoclonal antibodies were highly specific for human alfa-fetoprotein as determined by Western blotting. The affinity constants (K) of these Mab ranged from 10(6) to 10(9) l/mol.     

Plasmodium chabaudi: Expression of active recombinant chabaupain-1 and localization studies in Anopheles sp.

Plasmodium cysteine proteases have been shown to be immunogenic and are being used as malaria potential serodiagnostic markers and vaccine targets. Genes encoding two Plasmodium chabaudi cysteine proteases chabaupain-1 (CP-1) and chabaupain-2 (CP-2) were identified and further expressed in Escherichia coli. Solubilisation of recombinant CP-1 and CP-2 was achieved by decreasing the temperature of induction.Anopheles gambiae tissues infected with Plasmodium were analyzed by Western blotting using the anti-CP-1 antibody showing that CP-1 is only present in the A. gambiae midguts being absent from other infected mosquito biological material. Anti-CP-1 anti-serum recognized a 30 kDa band in P. chabaudi, Plasmodium berghei and Plasmodium yoelii lysates but does not recognize the recombinant CP-2 extracts suggesting high antibody specificity.     

Recombinogenic Telomeres in Diploid Sorex granarius (Soricidae,Eulipotyphla) Fibroblast Cells

The telomere structure in the Iberian shrew Sorex granarius is characterized by unique, striking features, with short arms of acrocentric chromosomes carrying extremely long telomeres (up to 300 kb) with interspersed ribosomal DNA (rDNA) repeat blocks. In this work, we investigated the telomere physiology of S. granarius fibroblast cells and found that telomere repeats are transcribed on both strands and that there is no telomere-dependent senescence mechanism. Although telomerase activity is detectable throughout cell culture and appears to act on both short and long telomeres, we also discovered that signatures of a recombinogenic activity are omnipresent, including telomere-sister chromatid exchanges, formation of alternative lengthening of telomeres (ALT)-associated PML-like bodies, production of telomere circles, and a high frequency of telomeres carrying marks of a DNA damage response. Our results suggest that recombination participates in the maintenance of the very long telomeres in normal S. granarius fibroblasts. We discuss the possible interplay between the interspersed telomere and rDNA repeats in the stabilization of the very long telomeres in this organism.     

Seasonal changes in chlorophyll distributions in Amazon floodplain lakes derived from MODIS images

To assess seasonal changes in phytoplanktonic chlorophyll distributions in Amazon floodplain lakes, a linear mixing model was applied to Moderate Resolution Imaging Spectroradiometer (MODIS) reflectance data acquired at four river stages: rising (April), high (June), decreasing (September), and low (November). The study area is located in a floodplain reach from Parintins (Amazonas) to near Almeirim (Pará). A three-end-member mixing model designed to uncouple three fractions [high suspended inorganic matter (ip), low inorganic suspended matter (w), and high chlorophyll a (Chl)] was tested in Lake Curuaí (1.5°S 55.43°W) based on field sampling done almost concurrently with satellite overpasses. During high water, phytoplankton patches are confined to lakes closer to terra firme under the influence of clear water inflow, whereas during the low and decreasing water stages, the patches are more evenly distributed over the floodplain.     

Changes in the canopy structure of the Mediterranean shrub Lavandula stoechas after disturbance

Abstract. This study attempts to show the dynamics of the canopy structure of the Mediterranean pioneer shrub Lavandula stoechas after man-made perturbation (i.e. grazing). The development of the vertical structure of the shrub was studied by harvesting the canopy of plants of 2–6 yr old in horizontal layers. The supportive biomass of the canopy was concentrated near the base at all ages. Leaf biomass was evenly distributed all over the vertical profile in 2- and 3-yr old plants. In 4-yr old plants it presented a maximum near the top of the canopy. For 5-yr old plants a structural transition started with leaf profiles showing a bimodal distribution. Leaf biomass predominated near the base in 6-yr old plants, suggesting that the transition was completed. Three canopy stages in the growth processes of the plant were recognized after the first year of growth: in the first one (from 2 to 3 yr old) both leaf and supportive biomass increased; in the second one (from 3 to 4 yr) leaf biomass remained stable and there was an increase in supportive biomass until the plants reached a ‘mature stage’, in 4-yr old plants; finally, in 5- and 6-yr old plants there was a decrease both in leaf and supportive biomass and plant structure showed evidence of senescence. Early transitions from seedling to 1-yr old plant and from this to 2- to 3-yr old plants were less obvious. The leaf/supportive biomass ratio always decreased with plant age, from 1.88 in seedlings to 0.01 in 6-yr old plants. Biomass density followed the pattern of supportive biomass, with an increase from 1.7 g/dm³ (2-yr old plants) to 2.4 g/dm³ (4-yr old plants). Thereafter, biomass density decreased to 0.6 g/dm³ (6-yr old plants).     

Long-Range Enhancer Interactions Are Prevalent in Mouse Embryonic Stem Cells and Are Reorganized upon Pluripotent State Transition

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A new role for PGRP-S (Tag7) in immune defense: lymphocyte migration is induced by a chemoattractant complex of Tag7 with Mts1

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7–Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target. Cells of either innate immunity (neutrophils and monocytes) or acquired immunity (CD4⁺ and CD8⁺ lymphocytes) can produce this complex, which confirms the close connection between components of the 2 branches of immune response.     

Effect of Salinity on Zinc uptake by Brassica juncea

Salinity is a major worldwide problem that affects agricultural soils and limits the reclamation of contaminated sites. Despite the large number of research papers published about salt tolerance in Brassica juncea L., there are very few accounts concerning the influence of salinity on the uptake of trace metals. In this study, B. juncea plants divided through soil sets comprising 0, 900 and 1800 mg Zn kg^?1, were treated with solutions containing 0, 60 and 120 mmol L^?1 of NaCl, with the purpose of observing the effect of salt on Zn uptake, and some physiological responses throughout the 90 days experiment. Increasing concentrations of NaCl and Zn produced a decline in the ecophysiological and biochemical properties of the plants, with observable synergistic effects on parameters like shoot dry weight, leaf area, or photochemical efficiency. Nevertheless, plants treated with 60 mmol L^?1 of NaCl accumulated striking harvestable amounts of Zn per plant that largely exceed those reported for Thlaspi caerulescens. It was concluded that salinity could play an important role on the uptake of Zn by B. juncea. The potential mechanisms behind these results are discussed, as well as the implications for phytoremediation of Zn on saline and non-saline soils.