Isolation of heparin-binding growth factors from bovine, porcine and canine hearts

Fresh bovine, porcine and canine hearts were homogenized and mitogens for mesoderm-derived cells were purified in three different steps. Extraction by two different ammonium sulfate precipitations was followed by cation-exchange chromatography and by heparin-Sepharose affinity chromatography. A heparin-Sepharose fraction from heart (eluted at 1.1 M NaCl) increased mitotic activity in serum-deprived cultures of porcine aortic endothelial and smooth muscle cells, and in human fibroblasts. This mitogenic activity is potentiated by heparin and inhibited by gamma-interferon. The heart mitogenic fraction showed one double peak on HPLC at A215 and one polypeptide band on SDS/PAGE. These peaks and bands were identical to those obtained from bovine brain. The heart acidic fibroblast growth factor (aFGF) showed a positive signal in Western blots using antibodies raised against brain aFGF. Gas-phase amino acid sequencing established that the mitogens were identical to aFGF and the N-terminally truncated aFGF. Extraction in the presence of a protease inhibitor (pepstatin A) produced a higher-molecular mass form of aFGF with a blocked amino terminus. Another mitogen, eluted at 1.6 M NaCl from heparin-Sepharose, reacted with polyclonal antiserum against human recombinant basic fibroblast growth factor (bFGF) and showed a 66% (12 from 18 amino acids determined by gas-phase sequencing) similarity with bFGF. This polypeptide increased the mitotic activity of the same cell lines but was more potent than aFGF.     

Resonance Raman spectroscopy of amicyanin, a blue copper protein from Paracoccus denitrificans

The copper binding site of amicyanin from Paracoccus denitrificans has been examined by resonance Raman spectroscopy. The pattern of vibrational modes is clearly similar to those of the blue copper proteins azurin and plastocyanin. Intense resonance-enhanced peaks are observed at 377, 392, and 430 cm-1 as well as weaker overtones and combination bands in the high frequency region. Most of the peaks below 500 cm-1 shift 0.5-1.5 cm-1 to lower energy when the protein is exposed to D2O. Based on the pattern of conserved amino acids, the axial type EPR spectrum, and the resonance Raman spectrum, it is proposed that the copper binding site in amicyanin contains a Cu(II) ion in a distorted trigonal planar geometry with one cysteine and two histidine ligands and an axial methionine ligand at a considerably longer distance. Furthermore, the presence of multiple intense Raman peaks in the 400 cm-1 region which are sensitive to deuterium substitution leads to the conclusion that the Cu-S stretch is coupled with internal ligand vibrational modes and that the sulfur of the cysteine ligand is likely to be hydrogen-bonded to the polypeptide backbone.     

Platelet count on slow induction to high altitude

Platelet counts were estimated at sea level in 50 lowlanders. They were divided at random in two groups (A and B) of 25 each. Group A went up by train/road transport to 3658 m, while group B reached the same height after 8 days of acclimatisation enroute. Platelet counts were estimated serially in both groups at high altitude. Symptoms of high altitude exposure were also recorded. No significant change in the counts was noted in either group and none became Symptomatic. All were brought back to sea level by air and deinduction studies carried out on days 1 and 4 of return. The importance of these findings in the light of our existing knowledge is discussed.     

Geminate recombination of CO in rabbit, opossum, and adult hemoglobins

The geminate recombination of CO with Hb following dissociation by a 10-ns laser pulse has been studied as a function of pH (9.2 and 7.0 without inositol hexaphosphate and 6.0 with inositol hexaphosphate) and temperature (5-35 degrees C). The hemoglobins studied included adult, Rothschild, rabbit, opossum, and carp. Despite significant differences in their structural and functional properties, the first four of these hemoglobins show similar trends in the yields, rates, and activation energies of the geminate recombination. The nature of the "cage recombination" in hemoglobin is discussed in the light of such findings. Neither a slow diffusion model nor a model based upon a specific non-heme binding site accounts for the observations.     

A note on some Eurotatoria from Panjab State,India

Among 35 eurotatorian species presently reported from Panjab State, India, ten species are new records to this region while eight are new records from N. W. India. A new synonym is proposed and biogeographical remarks are made.     

Nitrate reductase,nitrite reductase activities and leaf diffusive resistance in wheat under water deficit

The flag and second leaves of wheat showed physiological and biochemical differences in the normal and water stressed plants. Although the flag leaf had lower enzyme activities in the control plants, in stressed plants it could continue nitrate assimilation better than the second and other leaves. The flag leaf also exhibited a higher tendency to resume normal metabolic activity after the release of stress as is indicated by higher NRA, NiRa, lower r_ad and r_ab.     

Kinetics of reabsorption of nutrients in renal brush border membrane vesicles from rats with experimental ascending pyelonephritis

The uptake of nutrients was investigated in the renal cortical brush border membrane (BBM) vesicles at different stages of ascending pyelonephritis. There was significant difference (p less than 0.05) in the uptake of D-glucose, L-alanine, L-aspartate, L-lysine and L-proline 3 days postinfection and onwards in both right unobstructed and left obstructed experimental kidneys as compared to the sham operated control. The uptake of D-glucose, L-lysine and L-proline was found to be significantly decreased (p less than 0.05) during the course of infection. While uptake of L-alanine and L-aspartate increased (p less than 0.05) in early stages and decreased (p less than 0.05) in later stages of infection. The differential effect was attributed to the compensatory measure and different kinds of transport systems for different types of amino acids.