Early biochemical events in lymphocyte activation. I. Investigations on the nature and significance of early calcium fluxes observed in mitogen-induced T and B lymphocytes.

⁴⁵Ca²⁺ uptake was detected within minutes following addition of T- and B-cell² mitogens to mouse lymphocytes. The T-cell mitogens (Con A and PHA) gave an ~twofold increase in ⁴⁵Ca²⁺ uptake (representing an influx of ~ 130 amol per lymphocyte, corresponding to an increase in average cellular Ca²⁺ of ~0.95 mM). B-cell mitogens which gave the largest ⁴⁵Ca²⁺ uptake (~twofold) were purified LPS preparations from Salmonella minnesota R595 and Escherichia coli 0111:2125. The ⁴⁵Ca²⁺ uptake by rabbit splenocytes using specific anti-b4 allotype antiserum was comparable to that obtained with the two purified LPS preparations. A23187, in low nontoxic doses, gave an ~sixfold increase in ⁴⁵Ca²⁺ uptake with mouse T cells. The ⁴⁵Ca²⁺ uptake was modulated by cyclic nucleotides showing a “yin-yang” effect. The results suggest a possible entry of ⁴⁵Ca²⁺ from the extracellular medium through “gated Ca²⁺ channels” in the plasma membrane into the cytosol by passive diffusion. The Ca²⁺ may be sequestered in the mitochondria, and the excess Ca²⁺ is later effluxed into the extracellular medium. The fact that ⁴⁵Ca²⁺ uptake appears to be one of the earliest events occurring after ligand binding to the cell, together with the demonstration of a Ca²⁺-dependent glucose uptake and a requirement for extracellular Ca²⁺ for DNA synthesis, suggest that, as it is now known to function in many other cellular responses, Ca²⁺ may operate as a second messenger for lymphocyte activation.     

Human skeletal remains from Cossack,Western Australia

Human remains of a male individual from Cossack, northwestern Australia are described. Absolute dating is not possible but site geomorphology restricts the upper limit to 6500 B.P. Morphologically and metrically the skull differs from those of recent Western Australian male Aborigines, but it is very similar to that of Kow Swamp I and others included in the “robust” prehistoric Australian Aboriginal group (Thorne, 1977). The specimen is important as indicating the widespread nature and probable recency of a large, robust Australian Aboriginal population demonstrably different to recent populations.     

Relative growth rates of limb muscles in the diprotodont marsupial, Setonix brachyurus

The fore- and hindlimb muscles of 12 Setonix brachyurus joeys, aged 5 to 175 days postpartum, and four adults were dissected out and weighed. Individual muscles and muscle groups were analysed for absolute and relative growth changes. From comprising almost 59% of the total limb musculature at birth the forelimb muscles finally constitute just over 9% in the adult; the hindlimb muscles start at just over 41% and end at almost 91%. In both limbs, the extensor actions predominate in the proximal limb segment because of their propulsive functions, whereas in the distal segment the flexor muscles tend to be the larger because of their shock-absorbing and spring functions. During growth of the fore-limb there is a relative increase in the size of latissimus dorsi and triceps brachii and a decrease in the distal segment muscles; in the hindlimb the gluteal and hamstring muscles increase at the expense of the distal segment muscles. Specializations for speed include long distal hindlimb segments and proximally located muscle bellies. The above findings reflect the adaptations and changing locomotor patterns from birth to adult in the Quokka.     

Hemin feedback inhibition at reticulocyte δ-aminolevulinic acid synthetase and δ-aminolevulinic acid dehydratase

Addition of hemin (5–200 μM) to a rabbit reticulocyte iron-free incubation medium, resulted in a progressive inhibition of heme synthesis as measured by incorporation of (¹⁴C)-glycine. In contrast when (¹⁴C) δ-aminolevulinic acid incorporation into heme was studied, significant inhibition below that of the (¹⁴C)-glycine control only occurred with hemin concentrations greater than 100 μM. Hemin progressively inhibited cellular and mitochondrialδ-aminolevulinic acid synthetase activity, as well as cellular δ-aminolevulinic acid dehydratase activity. The results indicated that elevated levels of hemin initially control heme synthesis by feedback inhibition at the rate-limiting enzyme of heme synthesis, δ-aminolevulinic acid synthetase. Hemin inhibition of δ-aminolevulinic acid dehydratase is only significant for the entrire heme synthetic pathway when greater than one-third of this enzyme's activity is inhibited.     

A compact apparatus for vertical gel slab isoelectric focusing: resolution of rabbit anti-pneumococcal polysaccharide antibodies.

A compact apparatus for analytical and preparative vertical gel slab isoelectric focusing is described. The apparatus was extremely versatile, easy to use, and comparatively inexpensive. The apparatus was useful to compare the electrofocusing patterns of anti-pneumococcal polysaccharide antibodies from several different restricted and partially restricted rabbit responders.     

Engineering enzymes

Could we design and construct enzymes to catalyse any desired reaction? Compared with organic chemical catalysts, enzymes are highly specific and work in dilute aqueous solutions at ambient temperatures. Substrates are brought together from solution to precise orientations in the active site of an enzyme and the amino acid side-chains of the enzyme may assist catalysis by attacking or destabilizing substrate bonds. In principle, a novel enzyme could be constructed de novo or from pre-existing enzymes. Altering enzymes by recombinant DNA techniques offers most chance of success.