Standardization of the chromium-51 release, cell-mediated cytotoxicity assay: cryopreservation of mouse effector and target cells

Utilizing controlled cryopreservation techniques, we were able to standardize the 51Cr release cytotoxicity assay and thereby ensured reliable comparisons between results obtained on different days. Optimal conditions for freezing of both effector and target cells were quite similar. Dimethyl sulfoxide (DMSO) at a concentration of 7.5-10.0% was employed as the cryoprotective agent and cells were frozen at the rate of -1 degrees C/minute. The handling procedures for the cells before and after freezing were important. Factors affecting recovery of functional reactivity were related to toxicity of DMSO for the cells, the osmotic stress placed upon the cells as the DMSO was being removed after thawing, the handling temperature of the freshly thawed cells, and the susceptibility of cells to mechanical damage immediately after thawing. The recovery of lymphocytes after freezing was about 70%; the recovery of cytotoxicity was around 85%. Syngeneic cytotoxic reactivity induced by inoculation with the Moloney strain of murine sarcoma virus was cryopreserved, as were allogeneic cytotoxicity and natural cytotoxic reactivity. Multiple tests employing effector cells from the same frozen pool gave reproducible results; the standard error of the mean percent cytotoxicity was less than 1.5%. Cryopreserved target cells gave decreased day-to-day variability in susceptibility to lysis, since the same population of cells could be employed in each assay. These results demonstrated conclusively that we can now have a constant source of effector cells and target cells, which can be used from assay to assay as an internal standard.     

Effects of saponin capped triangular silver nanocrystals on the germination of Pisum sativum,Cicer arietinum,Vigna radiata seeds & their subsequent growth study

In this study, saponin capped triangular silver nanocrystals have been synthesised using fenugreek seed extract, where the extract acts both as a reducing and capping agent. X‐ray diffraction study confirms the purity and crystalline nature of the prepared nanocrystals and transmission electron microscopic study shows the triangular morphology with the average edge length of 72 nm, along with the atomic force microscopy study for the height or the width of the triangular nanocrystals. These nanocrystals have been investigated against a few pulses (seeds) such as Pisum sativum, Cicer arietinum and Vigna radiata for their effect on the germination as well as growth of root and shoot. Considering different concentration of silver nanocrystals solution, it has been found that 25 × 10⁻⁴ and 80 × 10⁻⁴ μg/ml are the minimum and maximum concentrations of silver nanocrystals, within this range, germination and subsequent growth of root and shoot are effective. The result shows significant positive influence on the growth of root and shoot of all seeds in comparison to those of unexposed control germination. Therefore, the result of this experiment has confirmed that the use of saponin capped silver nanocrystals enhances the germination and growth of plants.Inspec keywords: transmission electron microscopy, atomic force microscopy, nanofabrication, silver, nanostructured materials, X‐ray diffraction, crystal morphology, nanobiotechnology, botanyOther keywords: Pisum sativum, saponin capped nanocrystals, nanocrystals solution, atomic force microscopy, transmission electron microscopy, reducing agent, Vigna radiata seeds, Cicer arietinum, unexposed control germination, triangular nanocrystals, triangular morphology, x‐ray diffraction study, fenugreek seed     

Sediment Pickup on Streamwise Sloping Beds

This paper presents an experimental investigation on noncohesive sediment pickup under a unidirectional steady-uniform stream flow on streamwise steeply sloping (down slope and adverse) sedimentary beds. The characteristic parameters affecting the sediment pickup, identified based on the physical reasoning and dimensional analysis of the sediment particle movement under stream flow, are the transport-stage parameter, particle parameter, and geometric standard deviation of sediment particles. A large number of experiments (426 runs) were carried out in two long rectangular ducts (closed-conduit flow) with nine types of sediments (six uniform and three nonuniform sediments), having a variation of bed slope from ?15° (adverse slope) to 25° (down slope). In an open channel flow (laboratory flume study), the uniform flow is a difficult, if not impossible, proposition for a steeply sloping channel and is impossible to obtain in an adversely sloping channel. To avoid this problem, the tests were conducted with a closed-conduit flow. Measurements included flow discharge and sediment pickup rate. The bed shear stress for a particular run was computed considering side wall correction. The experimental data were used to determine the equation of sediment pickup function through a regression analysis. The equation is adequate to estimate sediment pickup not only on horizontal and mild slopes but also on steep and adverse slopes.     

Stability of magnetohydrodynamic stratified shear flows

Summary A study is made of the stability of a stratified shear flow in a perfectly conducting fluid in the presence of an external magnetic field aligned with the flow. A semi-circle theorem for the present hydromagnetic case is proved. The magnetic field is found to have a stabilizing effect on the flow. The Rayleigh-Taylor instability problem in a stratified conducting fluid is discussed. Finally, a study is made of the absorption of wave energy by the mean flow in the hydromagnetic case by considering a shear flow with an anti-symmetric velocity profile given byU=tanhz. Unlike the hydrodynamic case, it is found that, in the critical layer atU=0, the transfer of the wave energy to the mean flow occurs for any value of the Richardson number. This result implies again the stabilizing effect of the magnetic field on the shear flow.     

Synthesis, biochemical evaluation, and classical and three-dimensional quantitative structure-activity relationship studies of 7-substituted-1,2,3,4-tetrahydroisoquinolines and their relative affinities toward phenylethanolamine N-methyltransferase and the alpha2-adrenoceptor

7-Substituted-1,2,3,4-tetrahydroisoquinolines (7-substituted-THIQs) are potent inhibitors of phenylethanolamine N-methyltransferase (PNMT, EC 2.1.1.28), the enzyme involved in the biosynthesis of epinephrine. Unfortunately, most of these compounds also exhibit strong affinity for the alpha2-adrenoceptor. To design a selective (PNMT vs alpha2-adrenoceptor affinity) inhibitor of PNMT, the steric and electrostatic factors responsible for PNMT inhibitory activity and alpha2-adrenoceptor affinity were investigated by evaluating a number of 7-substituted-THIQs. A classical quantitative structure-activity relationship (QSAR) study resulted in a three-parameter equation for PNMT (PNMT pKi = 0.599pi - 0.0725MR + 1. 55sigmam + 5.80; n = 27, r = 0.885, s = 0.573) and a three-parameter equation for the alpha2-adrenoceptor (alpha2 pKi = 0.599pi - 0. 0542MR - 0.951sigmam + 6.45; n = 27, r = 0.917, s = 0.397). These equations indicated that steric effects and lipophilicity play a similar role at either active site but that electronic effects play opposite roles at either active site. Two binding orientations for the THIQs were postulated such that lipophilic and hydrophilic 7-substituents would not occupy the same region of space at either binding site. Using these two binding orientations, based on the lipophilicity of the 7-substituent, comparative molecular field analysis (CoMFA) models were developed that showed that the steric and electrostatic interactions at both sites were similar to those previously elaborated in the QSAR analyses. Both the QSAR and the CoMFA analyses showed that the steric interactions are similar at the PNMT active site and at the alpha2-adrenoceptor and that the electrostatic interactions were different at the two sites. This difference in electrostatic interactions might be responsible for the selectivity of THIQs bearing a nonlipophilic electron-withdrawing group at the 7-position. These QSAR and CoMFA results will be useful in the design of potent and selective (PNMT vs alpha2-adrenoceptor affinity) inhibitors of PNMT.