Identification and partial purification of a novel milk clotting enzyme from Onopordum turcicum

Summary Seeds, flowers and leaves of Onopordum turcicum were found to contain proteolytic enzymes able to coagulate milk. Extraction, concentration and identification of the operational parameters affecting the activity of the enzyme complex were followed by partial purification steps involving gel-filtration and ion-exchange chromatography. Milk clotting activity of the enzyme complex was tested in several steps of its purification and an increase of almost 200 fold was obtained. Molecular weight of the proteolytic enzyme fraction having the maximum activity was determined to be about 19000–24000. Isoelectric point (pI) of the enzyme complex with maximum activity was estimated to be in the range 3.3–3.7.     

Pistil Exudate Production and Pollen Tube Growth in Lilium longiflorum Thunb.

Exudate production in the pistil of Lilium longiflorum was studiedin relation to pollen tube growth, using scanning electron microscopy(SEM), transmission electron microscopy and light microscopy.In contrast with conventional fixation for SEM, during whichthe exudate of L. longiflorum largely washes away, the exudateremains present through freezing in case of cryo-SEM. Usingthe latter method we observed that exudate production on thestigma and in the style started before anthesis. Just underneaththe stigma the exudate was first accumulated at the top of eachsecretory cell, followed by a merging of those accumulationsas exudate production proceeded. Exudate is also produced bythe placenta. It was however not possible to determine whetherany of this fluid originated from the micropyle. Apart fromthe cell shape and the cuticle present in between the secretorycells, the ultrastructure of the secretory cells covering theplacenta was comparable to those of the stylar canal. The transferwall of the secretory cells of the placenta originated fromfusing Golgi vesicles but the endoplasmic reticulum seemed tohave an important role as well. After pollination the pollen tubes grew across the stigma andentered the style through one of the slits in the three stigmalobes. The pollen tubes grew straight downward through the styleand were covered by exudate. As the pollen tubes approachedthe ovary their growth was restricted to the areas with secretorycells. In the cavity the pollen tubes formed a bundle and theybent from this bundle in between the ovules towards the micropylarside. There they bent again to stay close to the secretory cells.After bud pollination the pollen tube growth was retarded. Laterarriving pollen tubes had a tendency to grow close to the secretorycells of the style, which resulted in a growth between thesecells and preceding pollen tubes. If there was still a littleexudate produced, it resulted in a lifting up of the pollentubes, out of the exudate. The relationship between exudateproduction and pollen tube growth is discussed. Both the speedand the guidance of the pollen tube seemed determined by theproperties of the exudate.Copyright 1994, 1999 Academic Press Cryo-scanning electron microscopy, exudate, Lilium longiflorum, lily, ovary, pollination, pollen tube growth, secretory cell, stigma, style     

Interaction between protein kinase C and regulatory ligand is enhanced by a chelatable pool of cellular zinc

At micromolar concentrations, zinc (Zn) and cadmium, but not other metals, greatly augmented binding of [3H]phorbol dibutyrate ([3H]PDBu) to protein kinase C (PKC) in cell homogenates and intact cells (in the presence of ionophore). Increased binding persisted for several hours. The heavy-metal chelating agent 1,10-phenanthroline completely reversed the increased [3H]PDBu binding in cells pretreated with 65Zn and ionophore and this was associated with a decline of about 20% in cell-associated 65Zn, suggesting that a relatively small pool of intracellular Zn acts on PKC. This may be a membrane-associated pool, since 65Zn readily bound to isolated erythrocyte inside-out membranes. Phenanthroline also partially inhibited binding of [3H]PDBu to PKC in untreated cells and extracts in a Zn-reversible manner. Therefore, cellular Zn appears to regulate the interaction of ligand with PKC. PKC bound to a Zn affinity column and was eluted by metal-chelator, confirming that Zn interacts directly with PKC.     

Morphofunctional indices of the effects of pyracryl on the organs of female genital system in rats

Morphofunctional indices of pyracryl (phosphate poly-2-pyridylethylmethacrilate) effecting reproduction system of Wistar rats have been studied. It has been shown that monthly administration of pyracryl in doses 1 mg/kg and 5 mg/kg lead to infiltration of mast cells and their pronounced degranulation and impairment of blood and lymph microcirculation. As far as gonadotropic effect is concerned, its parametres depend on the administration of pyracryl. Pyracryl in doses of 1 mg/kg stimulates folliculogenesis in ovaries, a dose of 5 mg/kg may cause expressed atresia of growing follicules and changes of oestrous cycle. It is suggested that biologically active substances of mast cells in female reproduction system play an important role in realization of gonadotropic effects of pyracryl.