Lectin-mediated agglutination of murine lymphoma cells. Cell surface deformability and reversibility of agglutination by saccharides

Agglutination of S49 mouse lymphoma cells by Ricinus communis I agglutinin can be reversed by the competing haptenic saccharide, lactose, soon after agglutination, but after further incubation in the absence of lectin the agglutination reaction could not be reversed by lactose and the cells remained as multicell aggregates. The irreversibility of S49 cell agglutination was time, temperature and lectin concentration dependent and its onset correlated with ultrastructurally observed deformation of adjacent cell surfaces and an increase in the proportion of adjacent cell surface areas in close apposition within multicell aggregates. Pretreatment of S49 cells with cytochalasin B or cytochalasin B plus vinblastine enhanced R. communis I agglutinin-mediated agglutination, while vinblastine alone and fluoride plus azide had essentially no effect. When drug-treated cells were agglutinated and then incubated in lectin-free drug-containing media for various times prior to lactose addition, the drug effects were more pronounced. Cytochalasin B alone or with vinblastine inhibited lactose reversal of S49 cell agglutination compared to the drug-free controls, while fluoride plus azide enhanced hapten reversibility. Electron microscopic analysis revealed that the onset of agglutination irreversibility correlated with cell surface deformation in the drug-treated cells. Cell aggregates that were more readily reversible by lactose (fluoride plus azide) were unchanged or less deformed, while S49 aggregates treated with cytochalasin B plus vinblastine were more deformed compared to controls without drugs. These experiments suggest a role for cell surface deformability as an important secondary effect during lectin-mediated cell agglutination of 849 lymphoma cells.     

Antifreeze proteins in the grubby sculpin,Myoxocephalus aenaeus and the tomcod,Microgadus tomcod: comparisons of seasonal cycles

Synopsis Antifreeze protein levels in the plasma of the grubby sculpin, Myoxocephalus aenaeus and the tomcod, Microgadus tomcod of Long Island coastal waters start to increase by November in anticipation of midwinter freezing conditions. Peak levels of antifreeze, as measured by the difference in plasma melting and freezing points, were detected in January for both species. The thermal hysteresis values reached 0.459°C in sculpin and 0.51°C in tomcod. Antifreeze peptides and glycopeptides start to disappear when water temperatures begin to rise and are at insignificant levels by late spring. Aspects of the seasonal cycle and the level of antifreeze activity were compared in three sympatric species (sculpin, tomcod, flounder); in two closely related but ecologically distinct gadids (tomcod, Atlantic cod); and within the genus Myoxocephalus.     

Lipopolysaccharide effects on sensitive and resistant variant chinese hamster ovary cell lines

Chinese hamster ovary (CHO · K1 · PRO) cell growth was inhibited by addition of a gram-negative bacterial lipopolysaccharide (LPS) to the cell culture medium. Growth inhibition began after three or four days of incubation, was dose-dependent up to a maximum at an LPS concentration of 500 μg/ml and was accompanied by cell shape changes and enhanced cytoplasmic vacuolization. Formation of bizarre CHO · K1 · PRO cell shapes and vacuole formation were most pronounced after seven days of incubation with LPS and could be observed by light and electron microscopy. An LPS-resistant cell population was obtained by intermittent in vitro exposure to high levels of LPS; these variant cells or clones derived from them failed to display growth inhibition in the presence of LPS. A clone from the LPS-resistant variant population showed altered cell properties compared to the parental cell line which included changes in cell morphology, adhesion, and endocytosis. Parental cells were markedly density-inhibited, whereas the variant clone exhibited considerable growth after confluency. The LPS-resistant variant cells showed a more elongated morphology than the parental line. No significant differences were observed between rates of detachment of parental and variant cells when sparse cultures of either line were removed from tissue culture dishes by ethylenediaminetetracetate (EDTA). However, at confluency approximately 100% of the variant cells versus 35% of the parental cells were removed by EDTA in one hour. Measurements of ¹²⁵I-ferritin uptake by parental and variant cells showed approximately twenty-fold and twofold increases, respectively, in uptake induced by LPS when compared to untreated control cultures.     

Haem iron intake in 12-36 month old children depleted in iron: case-control study.

OBJECTIVE: To compare the intakes of haem and non-haem iron in iron depleted and iron replete children. DESIGN: Case-control study. SETTING: Early Childhood Centres and a long day care centre in Sydney, Australia. SUBJECTS: Children aged 12-36 months depleted in iron and controls matched for age and sex. MEAN OUTCOME MEASURES: Iron status by using plasma ferritin concentration. A three day weighed dietary intake record completed by the parents. Risk factors for iron deficiency assessed by questionnaire. RESULTS: Fifty six iron depleted and 68 iron replete children participated. The average daily intake of haem iron was significantly lower in the iron depleted group (t = 2.392, P = 0.018); there was a tendency towards a lower average daily intake of non-haem iron (t = 1.724, P = 0.086) and vitamin C (t = 1.921, P = 0.057) for iron depleted children. Low intake of haem iron (< 0.71 mg/day) was significantly associated with iron depletion with an odds ratio fo 3.0 (P = 0.005). The proportion of iron depleted children who were given whole cows'' milk before 12 months of age was almost double that of iron replete children; multivariate analysis showed that both haem iron intake and age of introduction of cows'' milk were independently associated with iron depletion. CONCLUSIONS: The results of this study show that, in young children in developed countries, a lower haem iron intake is a major risk factor for iron depletion; the introduction of whole cows'' milk before 12 months is further confirmed as a risk factor. Parental education on nutrition should now focus on these two aspects of nutrition for infants and young children.