Decreased UDP-GlcNAc:Glycopeptideβ-2-N-Acetylglucosaminyltransferase II activity in a ricin-resistant mutant of baby hamster kidney (BHK) cells

The ricin-resistant mutant baby hamster kidney (BHK) cell line RIC^R21 is unable to make the sialylated bi- or triantennary complexN-glycans found in wild type cells and accumulates instead non-bisected hybrid structures containing three Man residues and one or two sialylated antennae (Hugheset al 1983, Carbohydr Res 120215-34). Specific assays forN-acetylglucosaminyltransferases I, II, III and IV were applied to Triton X-100 extracts of wild type BHK, RIC^R14 and RIC^R21 cells. It was shown that RIC^R21 cell extracts had a decreasedN-acetylglucosaminyltransferase II specific activity (17 to 27% of wild type values). It is suggested that in wild type cellsN-acetylglucosaminyltransferase II action proceeds quickly, leading to complexN-glycan synthesis, while in RIC^R21 cells potential substrates forN-acetylglucosaminyltransferase II move into the trans-Golgi compartment before the transferase can act, thereby leading to hybrid structures.     

Interrelationships of dietary protein level,aflatoxin B1 metabolism,and hepatic microsomal epoxide hydrase activity

In a continuation of studies on protein intake and aflatoxin B₁ (AFB₁) metabolism, weanling rats were fed semipurified diets containing either 20% casein or 5% casein for two weeks to determine the effect of dietary protein level on hepatic microsomal epoxide hydrase activity and AFB₁ metabolism in an effort to evaluate the role of protein intake on the formation and degradation of the reactive metabolite of AFB₁. Styrene oxide was used as substrate for epoxide hydrase since the hypothetical AFB₁ 2,3-epoxide (AFB-epox) cannot be synthesized because of its lability. Two groups of animals were fed 20% casein diets; one was fed ad libitum and the second was pair fed to the 5% casein group in order to control the effects of total feed intake. The depression of epoxide hydrase activities caused by the 5% casein diets was approximately equivalent to that previously seen with hepatic microsomal mixed function oxidase (MFO) activities with the identical protocol. Similarly, the metabolism of AFB₁ to AFQ₁ and AFM₁ was depressed by the 5% casein diets, with an increase in the production of chromatographically more polar material. The relationship of the MFO and epoxide hydrase activities to AFB₁ metabolism and formation of macromolecular adducts is discussed.     

The modulation of membrane fluidity by hydrogenation processes. III. The hydrogenation of biomembranes of spinach chloroplasts and a study of the effect of this on photosynthetic electron transport

A method is reported for the in situ modification of the lipids of isolated spinach chloroplast membranes. The technique is based on a direct hydrogenation of the lipid double bonds in the presence of the catalyst, chlorotris(triphenylphosphine)rhodium (I). The pattern of hydrogenation achieved suggests that the catalyst distributes amongst all of the membranes. The polyunsaturated lipids within the membranes are hydrogenated at a faster rate and at an earlier stage than are the monoenoic lipids.Whilst addition of the catalyst to the chloroplast causes an initial 10–20% decrease in Hill activity, saturation of up to 40% of the double bonds present can be accomplished without causing further significant alterations in photosynthetic electron transport processes or marked morphological changes of the chloroplast structure as observed in the electron microscope.     

Time course of osmotic adaptation and gill energetics of rainbow trout (Salmo gairdneri R.) following abrupt changes in external salinity

Summary The physiological and biochemical responses of rainbow trout (mean weight 250 g) to abrupt increases in salinity have been investigated. An initial crisis period lasting about 30 h was characterized by an increase in plasma and muscle ions, a rapid gill dehydration and a pronounced acidosis following a transient alkalosis. Mortality was low during this period. During the following days, gradual changes resulted in new steady state levels for most parameters examined.Analysis of adenylate pool (ATP, ATP/ADP ratio and energy charge) in the gills demonstrated an increased energy demand exhibiting two phases (4 h and 3 days), and a return to freshwater values. The gill respiration rate was constant during 3 days in sea water and decreased slightly later on. It was not influenced during the reverse transfer of seawater adapted fish into fresh water at the level of either isolated gills or perfused heads.     

Location ofMls locus on mouse chromosome 1

Linkage between theMls locus and the chromosome 1 markersDip-1 andald was detected using two sets of recombinant inbred strains. Linkage betweenMls andDip-1 was confirmed in the fifth and sixth backcross generations of an incipient congenic strain. The AKXL data indicate that the gene order isDip-1-ald-Mls. The recombination frequency betweenald andMls is estimated to be 0.07 ±0.05, based on the AKXL data. The recombination frequency betweenDip-1 andMls is estimated to be 0.18 ±0.04, based on all the available data.     

Prospectively randomised trial of proximal gastric vagotomy either with or without pyloroplasty in treatment of uncomplicated duodenal ulcer.

A consecutive series of 100 men with uncomplicated duodenal ulcer was randomly divided into two groups: one group of 52 underwent proximal gastric vagotomy (PGV), the other group (48) underwent PGV with pyloroplasty (PGVP). Preoperative peak acid output (PAOP) was measured in all patients. Those with a higher preoperative PAOP were significantly more likely to develop recurrent ulceration. Three patients developed recurrent ulceration after PGV and seven after PGVP. Dumping was both more common and more severe after PGVP than PGV. An overall satisfactory result was achieved in 92% after PGV and 81% after PGVP. We conclude that combining pyloroplasty with PGV has no appreciable advantages.     

Protein-lipid interactions and differential scanning calorimetric studies of bacteriorhodopsin reconstituted lipid-water systems

Bacteriorhodopsin has been reconstituted at various molar concentrations into liposomes of dimyristoyl- and also of dipalmitoylphosphatidylcholine. Differential scanning calorimetry indicates that as the protein concentration within the lipid bilayer increases, the cooperativity of the lipid phase transition is reduced, i.e. the transition is broadened, while the midpoint transition temperature remains virtually unchanged. Freeze-fracture electron microscopy of our preparation shows, in agreement with previous data from other laboratories, that extensive protein aggregation occurs when the liposome is cooled below the T_c transition temperature of the lipid. Laser flash photolysis measurements of protein rotation of the bacteriorhodopsin show, especially in the case of protein-rich recombinants, that protein aggregates exist even above T_c. The perturbation caused by the presence of bacteriorhodopsin in the lipid bilayer is similar to that produced by other intrinsic proteins. The difficulty of correlating the observed calorimetric enthalpy data with a simple concept of a ‘boundary lipid layer’ based upon consideration of a single isolated protein is discussed in view of the occurrence of protein aggregates both above and below T_c. It is concluded that the reduction of enthalpy is related to the number of lipids which solvate the protein aggregates within the protein-lipid patches and are thereby removed from the cooperative melting and enthalpy of the remaining regions of pure lipid.     

Proteolytic modification of mouse liver catalase

When catalase was immunoprecipitated from different subfractions of mouse liver homogenates, the enzyme which was obtained from extracts of the large granular fraction exhibited a lower molecular weight than that from either the cytosol or purified peroxisomal fractions, as judged by sodium dodecyl sulphate polyacrylamide gel electrophoresis. This modification of the enzyme could be prevented by the addition of proteolytic inhibitors to extraction buffers; and consequently, unmodified catalase was able to be purified in the presence of 5 mM iodoacetamide. Electrophoretic comparison of the catalases against standards of known molecular sizes indicated that the unmodified enzyme had a subunit mass approximately 2,000 daltons larger than the modified enzyme. The significance of these proteolytic modifications has been discussed in relation to the involvements of catalase and peroxisome turnover.