Changes in starch-bound lysophospholipids and lysophospholipase in germinating glacier and Hi amylose glacier barley varieties

Total starch, amylose content and amylose-included lipid phosphorus and lysophosphatidylcholine (LPC) were measured in normal Glacier (G) and Hi Amylose Glacier (HA) barley varieties during germination. From days three to six, alkaline and acidic lysophospholipase (LPL) activities in the starchy endosperm were measured and the distribution of these activities between a soluble and particulate form determined. During germination the amylose content of the starches increases as the total starch levels decline. The starch-bound LPC and lipid phosphorus disappear at the same rate between days three and six in both barley varieties, indicating no discrimination among the different lipid-included amylose population for degradation. However, both lipid phosphorus and LPC disappear more rapidly in the G than in the HA variety. This is presumably due to the slightly larger content of LPC per mg amylose of the G than of the HA variety, equivalent to 134 and 150 anhydroglucose residues per lipid molecule in G and HA, respectively. There is no increase in starch-bound lipid phosphorus or LPC expressed as nmol of phosphorus or LPC per mg amylose as amylose content declines, indicating no selective resistance of lipid-included amylose to degradation. The alkaline and acidic LPC activities in each variety increase 2–4-fold between days four and five. In both varieties ca 30% of the acidic LPL and ca 50–60% of the alkaline LPL is particulate from days three to six. No correlation can be made between the content of amylose or amylose-included lipid and particulate LPL activity. However, the possibility that particulate LPL activity is associated with specific populations of residual amylose-included lipid molecules cannot be excluded.     

Isolation and characterization of a 23 kDa protein essential for photosynthetic oxygen evolution

A 23 kDa protein has recently been demonstrated to participate in photosynthetic oxygen evolution by reconstitution experiments on inside-out thylakoid vesicles (Åkerlund H-E, Jansson C and Andersson B (1982) Biochim Biophys Acta 681:1–10). Here we describe the isolation of the 23 kDa protein from a spinach chloroplast extract using ion-exchange chromatography. The protein was obtained in a yield of 25% and with less than 1% of contaminating proteins. The ability of the protein to stimulate oxygen evolution in inside-out thylakoids was preserved throughout the various fractionation steps. The isolated protein was highly water soluble and appeared as a monomer. Its isoelectric point was at pH=7.3. The amino acid composition showed a high content of polar amino acids, resulting in a polarity index of 49%. The isolated protein lacked metals and other prosthetic groups. Its function as a catalytic or regulating subunit in the oxygen evolving complex is discussed.     

Outbreak of Giardiasis: Effect of a New Antiflagellate Drug,Tinidazole

We describe here an intensive outbreak of mostly symptomatic (90%) Giardia lamblia infestation in a Swedish student group visiting the U.S.S.R. A new antiflagellate drug, ethyl (2-(2-methyl-5-nitro-1-imidazolyl)ethyl) sulphone, tinidazole, Pfizer, was given in a dosage of 150 mg twice daily for seven days to 10 healthy volunteers and to 24 students infested with G. lamblia. The drug was found to be effective in curing giardiasis and in eradicating G. lamblia from the intestinal tract. All the students with symptomatic giardia infestation became free from gastrointestinal disturbance, usually soon after treatment was started. None of the 24 students had G. lamblia in their stools after tinidazole treatment was discontinued or at follow-up. No side effects of the drug were seen and all of the subjects tolerated it very well.     

Genetic transformation of apple (Malus pumila Mill.) using a disarmed Ti-binary vector

The disamed Ti-binary vector pBIN 6 in Agrobacterium tumefaciens has been used in leaf disc transfomations to produce transgenic apple (Malus pumila Mill.) plants with a nomal phenotype except for a somewhat reduced capacity to root. The presence of the genes for nopaline synthase and neomycin phosphotrans ferase (conferring kanamycin resistance), inserted into the host genome by the vector, was confirmed by Southern blot analysis, the detection of nopaline synthase activity and rooting in the presence of the antibiotic.The nopaline synthase gene continued to be expressed in glasshouse-grown plants several months after removal from in vitro growth conditions.     

Purification of two muscle enzymes by chromatography on immobilized ferric ions

Two enzymes, glycogen phosphorylase and lactate dehydrogenase, were purified simultaneously in a single step. Ferric ions immobilized on a chelating gel were used as the adsorbent. Adsorption and desorption steps were accomplished by changes in buffer composition. The recoveries were better than 80% and the capacities were about 5 mg of protein per milliliter of adsorbent. The procedure worked well both on a small and on a preparative scale. The homogeneity of the purified enzymes was checked by FPLC.     

Expression cloning and regulation of steroid 5 alpha-reductase, an enzyme essential for male sexual differentiation

The conversion of testosterone into the more potent androgen, dihydrotestosterone, catalyzed by the enzyme steroid 5 alpha-reductase, is required for the differentiation of male external genitalia. Here, we report the isolation of cDNA clones encoding the rat steroid 5 alpha-reductase using expression cloning in Xenopus oocytes. DNA sequence analysis demonstrates that the liver and ventral prostate forms of steroid 5 alpha-reductases are identical hydrophobic proteins of 29 kDa. The amount of steroid 5 alpha-reductase mRNA in liver increased in response to castration, but remained unchanged in the prostate. Testosterone administration to castrates induced expression of mRNA in the prostate but had no effect on liver. The data suggest that the steroid 5 alpha-reductase gene is differentially regulated by testosterone in androgen-responsive versus non-responsive tissues.