The use of two populations of hepatocytes with different triacylglycerol contents as a model to study the accumulation of liver lipid in the laying hen.

(1) A method has been developed to separate hepatocytes, isolated from laying hens, according to their densities, using discontinuous density-gradient centrifugation on Nycodenz. (2) The hepatocytes recovered from the interface of the 5% and 10% Nycodenz layers were rich in triacylglycerol and were termed 'fatty' hepatocytes: 'non-fatty' hepatocytes were obtained from the interface of the 15% and 30% Nycodenz layers and contained less than one-quarter as much triacylglycerol. (3) 'Fatty' hepatocytes incorporated radiolabelled glucose and glycerol into total lipid at more than twice the rate of 'non-fatty' cells: the corresponding increases in the incorporation of radiolabelled choline and valine into phospholipid and protein respectively were smaller and not statistically significant. (4) A higher proportion of glycerol and glucose incorporated into total lipid was found to be phospholipid in the 'non-fatty' hepatocytes. (5) A higher proportion of radiolabelled lipid or protein formed from glycerol or valine respectively was secreted into the medium by the 'non-fatty' hepatocytes. (6) The use of these hepatocytes as a model to study fatty liver syndromes is discussed.     

Analysis of the sheep MHC using HLA class I,II, and C4 cDNA probes

Four cDNA probes for the human major histocompatibility complex (MHC) were used to investigate the sheep MHC, in conjunction with serological typing for ovine lymphocyte antigen (OLA). Lymphocytes from a family (two parents and five offspring) of Romanov sheep were subjected to genomic DNA digestion by the restriction endonuclease Eco RI, followed by gel electrophoresis. A single Southern blot representing all seven individuals was then consecutively hybridized with the class I, alpha-DC, beta-DR, and C4 probes, which were originally designed to identify HLA class I, class II (DC and DR), and C4 products, respectively. Using each of the three class I/class II probes, several bands showing DNA polymorphism were detected. The segregation of these bands in the five offspring exactly paralleled the OLA haplotype segregation established by serological typing. A further eight individuals carrying haplotypes which were phenotypically identical to those in the above-mentioned family showed bands in the corresponding positions when tested with the same three probes. Using the C4 probe, no polymorphism was detected in these fifteen individuals.Abbreviations used in this paper MHC major histocompatibility complex - OLA ovine lymphocyte antigen - kbp kilobase pair(s) - MLR mixed lymphocyte reaction - RFLP restriction fragment length polymorphism     

Availability,uptake and regeneration of phosphate in mesocosms with varied levels of P deficiency

A population of cyclomorphic Bosmina coregoni was studied in Lake Östersjön, southwestern Sweden and results from field samples collected in 1988, 1989, 1990 and 1991 are presented. Animals collected in summer have remarkably higher carapace and prolonged antennule compared to what we call the normal morph. In 1991 the extreme morph reach its maximum body length, body height and antennule length in July to September. The occurrence of the extreme morph coincide with the hatching of the predaceous cladoceran Leptodora kindtii.The two morphs fluctuate in abundance and in relation to each other. In early spring only the normal morph occurred in the samples followed by a period of about two months when the two morphs were found together, in July only the extreme morph was found. In September the two morphs were again present in the lake. As has been shown for other cladoceran, the conspicuous carapace and antennule could be an adaptive response that decreases mortality due to invertebrate predation. Spectacular features like these are likely also accompanied by some sort of costs.     

Activation of the first component of human complement (C1) by antibody-antigen aggregates.

The activation of subcomponents C1r and C1s in the first component of complement, C1, when bound to antibody-antigen complexes was investigated. Activation was followed both by the splitting of the peptide chains of subcomponents C1r and C1s and by the development of proteolytic activity. For the maximum rate of activation to occur, all components must be present in approximate molar proportions of antibody: C1q:C1r:C1s of 13:1:5:5. For activation of subcomponent C1s, subcomponents C1r or C1r, but not C1r inactivated with iPr2P-F (di-isopropyl phosphorofluorideate), are effective. For activation of subcomponent C1r, subcomponents C1s, C1s or C1s inactivated with iPr2P-F are effective. Subcomponent C1s is activated by C1r, and C1r is activated autocatalytically, probably through the formation of an intermediary C1r. in which the peptide chain is unsplit but a conformational change caused by interaction with the other components has led to the formation of a catalytic site able to split subcomponent C1r to C1r.     

Cell free synthesis of some storage protein subunits by polyribosomes and RNA isolated from developing seeds of pea (Pisum sativum L.)

Polyribosomes which have template activity in the wheat germ system have been isolated from developing pea seeds. Some of the translation products have identical mobilities to the vicilin and legumin subunits by SDS-PAGE. Certain products were specifically immunoprecipitated with antisera prepared against purified vicilin and legumin fractions. Various RNA fractions including poly A-rich RNA have also been isolated from polyribosomes and shown to direct the synthesis of polyripeptides whose properties are similar to the storage protein subunits. The results are discussed in relationship to other investigations with seed storage protein biosynthesis in vitro.Abbreviations DTT dithiothreitol - SDS-PAGE SDS-polyacrylamide gel electrophoresis - TCA tricarboxylic acid     

Combination v. sequential therapy with melphalan, 5-fluorouracil and methotrexate for advanced ovarian cancer.

The results of a national clinical trial to compare combination and sequential chemotherapy for stage III or IV ovarian cancer are reported. Of the 253 patients from 16 centres across Canada who were admitted to the trial 13 were excluded from the analysis. All the patients were observed for 2 to 5 years from entry into the trial. There were no differences in response to therapy or in survival between the patients treated with melphalan followed by 5-fluorouracil and then by methotrexate in high dosage and the patients treated with the same agents in combination. Patients with minimal residual disease after resection of stage III ovarian cancer had a good prognosis. Other favourable prognostic factors were age (less than 55 years), performance status (90% or 100% on the Karnofsky scale) and histologic grade of the tumour.     

The relationship between the size of mitochondria and the intensity of light that they scatter in different energetic states

The intensity of light scattered at 90° to the incident beam and the effective hydrodynamic radii of mitochondria incubated under a variety of conditions have been measured. Addition of high concentrations of uncouplers to respiring mitochondria resulted in a decrease in scatter which was not due to swelling. Addition of valinomycin to mitochondria depleted of substrate in K⁺-free medium produced an increase in scatter that was not due to shrinking. It is concluded that changes in the intensity of scattered light are not reliable indices of changes of volume of mitochondria, and that changes in conformation with changes in metabolic state dominate changes in light scatter. A molecular mechanism for the effect of metabolic state upon the scattered intensity is suggested.