Primary in vitro antibody formation by blood leucocytes of a subhuman primate

Blood leucocytes from humans and two species of marmosets, S. fuscicollis and S. oedipus oedipus, were cultured in vitro with sheep red blood cells. Leucocyte cultures from 16 of 20 S. fuscicollis marmosets produced significant numbers of plaque-forming cells (PFC) as assayed by the Jerne procedure. Under the same conditions, none of 15 S. oedipus oedipus cultures and only 1 of 15 human blood cultures responded. A kinetic study of the response with S. fuscicollis leucocytes revealed peak 19S and 7S PFC to appear on Day 9 of culture. It is suggested that two factors are acting synergistically in the successful development of antibody-forming cells from leucocytes of S. fuscicollis marmosets: (a) the innate immune response capabilities of the blood lymphocytes and (b) the natural hemopoietic chimerism that occurs in this species.     

Therapeutic effects of electromagnetic fields in the stimulation of connective tissue repair

The therapeutic effects of electric and magnetic fields have been studied largely for their promotion of connective tissue repair. The most widely studied application concerns bone repair and deals with acceleration of the healing of fresh fractures, delayed and non-unions, incorporation of bone grafts, osteoporosis, and osteonecrosis. More recently the effects of these fields upon the repair of cartilage and soft fibrous tissues have been described. In all these experimental systems and clinical applications an acceleration of extracellular matrix synthesis and tissue healing has been observed. A degree of specificity, in terms of the parameters of applied energy and biological response, is hypothesized.     

Evidence for cooperative effects in the binding of polyvalent metal ions to pure phosphatidylcholine bilayer vesicle surfaces

An internal NMR monitor for the study of lanthanide ion (Ln³⁺) binding to phospholipid bilayer membranes has been developed. The dimethylphosphate anion, DMP^?, forms labile complexes with Ln³⁺ in aqueous solution and in solutions also containing bilayer dispersions. The hyperfine shift in the DMP^? resonance induced by Pr³⁺ ions has been used to determine the overall thermodynamic formation constants for the Pr(DMP)²⁺ and Pr(DMP)₂⁺ complexes: 81 (M^?1) and 349 (M^?2) at 52°C; the limiting hyperfine shift (³¹P) at 52°C is 91.5 ppm downfield. These parameters, applied to the observed DMP^? hyperfine shift in the presence of the membrane, establish both the free Pr³⁺ concentration and the amount of Pr³⁺ bound to the phospholipid surface. Extensive data for the binding of Pr³⁺ to the outer surfaces of sonicated vesicles yield a limiting hyperfine shift per Pr³⁺ of 181.6 ppm downfield for the dipalmitoylphosphatidylcholine ³¹P resonance at 52°C, clearly demonstrating that the binding stoichiometry is two DPPCs per Pr³⁺. A Hill analysis indicates that the binding data are more anti-cooperative than a realistic Langmuir isotherm, yet more cooperative than a Stern isotherm incorporating electrostatic considerations at the Debye-Hückel level. Fittings to specific models lead to a cooperative model in which tense (T) sites, with low affinity for Pr³⁺, present in the absence of metal ions, quickly give way to relaxed (R) sites (two DPPCs per site), with much higher affinity for Pr³⁺, as the amount of Pr³⁺ bound to the surface increases. The intrinsic equilibrium constants for the binding of Pr³⁺ to DPPC vesicles are 2 M^?1 and 3 000 M^?1 for the T and R sites, respectively, at 52°C. The distribution coefficient between these sites ([R]/[T]) in the absence of Ln³⁺ is 0.14 at 52°C. We picture the binding site conversion as a head-group conformational change involving mostly the choline moiety. Sketchy results for binding on the inside vesicle surface indicate that the overall affinity for Pr³⁺ is significantly greater and suggest that the site stoichiometry may be different.     

Preliminary analysis of drinking from seawater sources by free-ranging rhesus macaques (Macaca mulatta)

Rhesus macaques on Key Lois Island were observed drinking seawater that flowed into four holes they had excavated in the sand. Data were gathered to determine the salinity (TDS) and pH levels of the water and which animals were using the holes. Average TDS level (4,506.8 ± 1,750.8), but not pH (8.1 ± .30) level, of water from the holes differed from the surrounding seawater (TDS = 29,000, pH = 8.0). There were significant variations in TDS and pH levels of water between holes. A total of 249 drinking and 11 digging bouts were observed. Adult females drank and dug most often (46.9% of total bouts). Of the 260 total drinking and digging bouts, 76.1% (N = 198) were concentrated at one hole. This hole had the lowest average TDS level (3,714.2 ± 1,504.4) and one of the highest average pH levels (8.1 ± .29). Age/sex class differences in drinking bout frequencies may have been due to differential social status. We suggest that the holes were excavated to overcome a temporary shortage of provisioned water. © 1993 Wiley-Liss, Inc.     

Antibodies specific for gp40 inhibit cell-cell adhesion by cross-linking the protein on the surface of Dictyostelium purpureum

We have previously suggested a role for gp40 in cell-cell adhesion in Dictyostelium purpureum from the fact that antibodies specific for this protein inhibited adhesion in an in vitro assay [Springer: Dev Biol 133:447–455, 1989]. To further confirm this role mutants lacking the protein were isolated and characterized. To our surprise, the mutants had normal adhesive properties both in vitro and in situ. These results lead us to the conclusion that gp40 is not necessary for the cell-cell adhesions observed and may not be a molecule which directly participates in these adhesions. When studied further, we found that adhesion-inhibitory antibodies were only effective as divalent IgG. Monovalent Fab fragments of the same antibodies could not inhibit adhesion. The inhibitory antibodies also caused the cells to remain rounded and incapable of attaching to plastic surfaces. We conclude that when divalent antibodies specific for gp40 cross-link this protein on the cell surface a cytoskeletal change prevents them from attaching to substratum as well as to other cells, thereby inhibiting cell-cell adhesion. We suggest that an alternative mechanism for inhibition of cell-cell adhesion by divalent antibodies exists and should be considered before proposing a direct role for a protein in adhesion.     

Synthesis of a fluorescent 2′3′-dideoxycytosine analog,tCdd

The pathway leading to the preparation of a novel tricyclic 2′3′-dideoxycytosine analog, tCdd (1) is reported. A protected 2′3′-dideoxyribose prepared from l-glutamic acid was coupled to a silylated fluorescent base to yield a mixture of the α- and β-anomers of the 2′3′-dideoxyribonucleoside of 1,3-diaza-2-oxophenothiazine, tCdd (1). The fluorescent base analog retains a high fluorescence emission over a large pH range and should be useful in a variety of probe applications.