Serological survey for canine leptospirosis in the Pretoria area.

Canine serum samples (n = 400) from the Pretoria area were tested for Leptospira antibodies, using the microscopic agglutination test. The prevalence of antibodies (inconclusive and positive titres) was 1.5%. Reactions were only against Leptospira interrogans serovars tarassovi and pyrogens. Leptospirosis does not appear to be an important canine disease in the Pretoria area.

A serological comparison of complement fixation reactions using Brucella abortus and B. melitensis antigens in B. abortus infected cattle.

Brucella abortus and B. melitensis antigens were used in parallel on the National Standard Brucella abortus antiserum and on field sera coming from cattle where practically exclusively B. abortus biotypes 1 and 2 have been isolated over the last 11 years. With the National Standard serum the titres to B. melitensis were consistently lower than those to B. abortus antigen. Most were 1 dilution (twofold) lower. Although a similar trend was seen with the field sera, there were 7/346 sera which had twofold or higher titres to B. melitensis antigen. Although this may be due to the vagaries of the test it also warrants closer investigation of the animals concerned to see whether M-antigen predominant Brucella biotypes are possibly present. The use of the dual antigens could identify herds which are infected only with A-antigen predominant brucellae but would not be reliable for classifying individual animals.

A serological survey for brucellosis in buffalo (Syncerus caffer) in the Kruger National Park.

A serological investigation was undertaken to determine the prevalence of brucellosis titres in buffalo in the Kruger National park. A total of 406 samples were collected over a period of one year. The rose bengal and the complement fixation tests were used in the investigation as these tests are routinely used for cattle sera and have proved to be reliable. In the females, 12.6% adult, 10.7% sub-adult and 3% juvenile animals reacted positively to the tests. In the males, 15.1% adults, 10.6% sub-adults and 5.3% juveniles were recorded as positive.

Beta-chain gene rearrangements and V beta gene usage in DPw2-specific T cells.

The diversity of human T cell receptor beta-chain gene rearrangements and variable region gene usage in the T cell response to a single allogeneic class II HLA gene product has been investigated. Nine clones of cytotoxic T lymphocytes (8.2 to 8.10) that are specific for the class II specificity DPw2 were analyzed for their T cell receptor beta-chain gene rearrangements using a constant-region probe. A minimum of seven different clonotypes were present in this panel of clones. The beta-gene expressed by one clone, 8.9, was isolated and the variable (V), diversity (D), and joining (J) segments were sequenced. The sequence of the V beta 8.9 segment is identical to the V beta 14 sequence, and is joined to D beta 1.1 and J beta 1.1 segments. Northern analysis revealed that three of the eight clones analyzed, 8.5, 8.7, and 8.9, expressed a 1.3 kb transcript that hybridized with the V beta 8.9 probe, indicating that these clones were using the same V beta gene (these clones shared common DNA rearrangements). The remaining five clones did not express V beta 8.9. Southern analysis of DNA obtained from a DPw2-specific tertiary mixed lymphocyte reaction bulk culture from which the clones were derived showed a prominent rearrangement of the V beta 8.9 gene that was indistinguishable from those observed for clones 8.5, 8.7, and 8.9. This prominent rearrangement of V beta 8.9 was not observed in DNA obtained from normal peripheral blood lymphocytes. These results suggest that although the number of V beta genes which can contribute to a DPw2 specificity may be relatively large, only a limited number of clonotypes ultimately predominate in the response to certain class II HLA antigens.

Abnormality of cultured muscle and Schwann's cells in familial lipid neuromyopathy. Muscle corrected by neural influence.

Muscle and Schwann's cell cultures were established from a family (mother and two sons) with non-carnitine deficient neuromyopathy. Electron microscopy of noninnervated cultured muscle showed poorly matured muscle fibers containing large "mushy" mitochondria, lipid droplets, abundant multilaminated inclusions, and dense-core dark osmiophilic bodies. Parallel innervated muscle fibers (cocultured with normal-rat motor neurons for two to three weeks) were well cross-striated, had well-developed T tubules and sarcoplasmic reticulum, and none of the abnormalities of aneural muscle cultures. Cultured Schwann's cells were normal by light microscopy, but had ultrastructurally abnormal mitochondria, lipid droplets, dark osmiophilic granular inclusions, and numerous "foamy" vacuoles. Those studies demonstrated (1) intrinsic muscle and Schwann's cell defects because of reproduction of abnormalities in culture, (2) normal muscle cell responsiveness to innervation reflected by development of contractions and longer survival in culture, and (3) beneficial influence of a neural factor(s) on the endogenous muscle cell abnormality.

Biochemical and morphological effects of 20,25-diazacholesterol on cultured muscle cells.

Effects of 20,25-diazacholesterol (DAC), a myotonia-inducing drug, were evaluated on certain biochemical and morphological properties of embryonic rat muscle cells grown in tissue culture. During DAC treatment, muscle fibers exhibited spontaneous contractions that changed from coarse twitches to finer fibrillation movements, The ultrastructural alterations produced by DAC were smeared Z-lines, disorganized myofibrils, occasional honeycomb appearance of membranes and large vacuoles connected to zipper-like structures. Biochemically, a microsomal fraction prepared from DAC-treated cells (compared to that of normal cells) showed a 30-45 per cent decrease in the isoproterenol-enhanced and the NaF-enhanced adenylate cyclase activity. however, the beta-adrenergic receptors, through which isoproterenol activates the enzyme, showed no change in density or affinity as judged by the binding of [125I]iodohydroxybenzylpindolol. That indicated that DAC treatment caused an uncoupling of beta-receptor-adenylate cyclase interaction. Guanylate cyclase and cyclic GMP-phosphodiesterase were both markedly increased in DAC-treated cells, indicating a greater turnover of cyclic GMP. Binding of [3H]concanavalin A to DAC-treated muscle membranes was decreased 20-40 per cent. The data indicate that DAC exert a direct influence on muscle fibers, affecting their functional, biochemical and morphological properties.

Human schwann cells in tissue culture: histochemical and ultrastructural studies.

Schwann cells cultures were established from 40 human peripheral nerves that underwent biopsy according to an explant-reexplantation technique that markedly reduces non-Schwann cells and achieves cultures greatly enriched in Schwann cells suitable for a wide range of studies. Described are the growth characteristics and histochemical, ultrastructural, and ultrastructural-cytochemical patterns of normal human cultured Schwann cells from 12 patients with demonstrable peripheral nerve abnormality. These findings serve as a basis of comparison when seeking abnormalities of Schwann cells grown from peripheral nerves of patients with putative dysschwannian neuropathies.

X-linked recessive congenital muscle fiber hypotrophy with central nuclei: abnormalities of growth and adenylate cyclase in muscle tissue cultures.

Muscle cells in cultures established from biopsy specimens of two children with an infantile-fatal form of X-linked recessive muscle fiber smallness with central nuclei showed an unusual ability to proliferate through numerous passages. Ultrastructurally, the cultured muscle fibers appeared very immature even after several weeks. The nuclei were large, the number of ribosomes was greatly increased, the myofibrils remained unstriated, and glycogen was accumulated in large lakes. The plasmalemma bound concanavalin A, alpha-bungarotoxin, and ruthenium red normally, but with tannic acid it did not show the dark binding of mature fibers. Biochemically, in the cultured muscle fibers, beta-adrenergic receptors were quantitatively normal. The level of adenylate cyclase in membranes was less than in cultured normal muscle; this defect could be responsible for impaired control mechanisms resulting in the other abnormalities observed.

Correction for the inherent error in optical density readings.

Except at very low levels, uncorrected photometric determination of bacterial cell densities showed a decreasing proportionally to actual cell density or dry weight. A standard curve was prepared to convert photometric readings to truly proportional optical density values. With one dry weight determination, optical density values may be converted to absolute dry weight values.