Genetics of New Crop Genus Limnanthes I. Five Morphological Marker Loci in L. alba × L. gracilis Progenies

New crop development often depends on a few essential traits such as nonshattering of seed at maturity and pollination control. In order to domesticate Limnanthes alba (meadowfoam), as a source of industrial oil, we performed genetic analysis of crosses between L. alba and its closest relative Limnanthes gracilis var. gracilis which shows higher selfing ability and nutlet abscission. Differences in pubescence, nutlet abscission, and seedling pigmentation were each found to be governed primarily by a single locus; two interacting loci appeared to control capsule dehiscence. Segregation distortion was detected for pigmentation in F₂ and F₃ populations, and for pubescence in several F₃ families. Loci controlling pubescence, nutlet abscission, and capsule dehiscence resided in a single linkage block which assorted independently of the locus controlling pigmentation.     

Detection of equine antiplatelet and antineutrophil antibodies by enzyme-linked immunosorbent assay

An enzyme-linked immunosorbent assay (elisa) was standardised and applied for the detection of anti-platelet and antineutrophil antibodies using a heterologous system consisting of equine platelets or neutrophils and antisera raised in rabbits. The standardised technique consisted of using Immulon type 3 plate, I per cent gelatine as a blocking solution, poly-L-lysine buffer as a coating solution, unfixed antigen, 90 μl test serum, horseradish peroxidase conjugated antibody and o-phenylenediamine dihydrochloride as a substrate. The number of unfixed platelets or neutrophils required for optimum detection of antibodies was 250,000 per well. Unfixed cellular antigens were as good as their extracts and superior to paraformaldehyde-fixed antigens in detecting specific antibodies. Microtitre plates coated with platelet or neutrophil antigens could be stored at 4° and −70°C for four to five weeks without significant loss of antigenicity. The ELISA was very sensitive in that antiplatelet antibody was detected up to a titre of 1:204,800 and antineutrophil antibody to a titre of 1:51,200. Some cross-reactivity (1:1600) was detected in antiplatelet and antineutrophil sera for neutrophil and platelet antigens, respectively. Platelet-associated antibody was also detected in extracts from platelets pretreated with 1:2 and 1:8 dilutions of antiplatelet serum. Standardised elisa detected antiplatelet antibodies in nine and antineutrophil antibodies in three of 100 isologous equine blood typing sera.     

Use of flow cytometry for determination of differential leukocyte counts in bovine blood

A flow cytometric method was developed to perform differential leukocyte counts on bovine blood. Blood specimens from 50 healthy Holstein cows were analyzed by use of a flow cytometer. The method entailed diluting blood with phosphate-buffered, hypotonic saline solution containing acridine orange, and performing a step-wise, 3-parameter analysis on the bases of cell size, cellular granularity, and granulocyte fluorescence. Initially, proportions of monocytes, granulocytes, and lymphocytes were determined by creating appropriate windows on dot plots of cell size (determined by forward light scatter) vs cellular granularity (determined by the logarithm of side light scatter). Eosinophils were resolved by analysis of granulocytes as dot plots of logarithms of green vs red fluorescence ascribed to acridine orange. Proportions of eosinophils and neutrophils were computed from data so generated. Microclumps of platelets spuriously affected counts of some granulocytes, particularly eosinophils. Differential leukocyte counts determined by flow cytometry generally compared favorably with those obtained by use of the conventional microscopic method, using Wright-stained blood films. Mean neutrophil and eosinophil counts determined by the 2 methods did not differ significantly, but lymphocyte counts determined by flow cytometry were significantly higher than those determined by microscopy (P less than 0.01). Correlation coefficients for counts of neutrophils, eosinophils, and lymphocytes determined by the 2 methods ranged from 0.519 to 0.833. Correlation between monocyte counts was low (r = 0.147), although mean monocyte counts determined by the 2 methods did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hybrid cell line from a cloned immunoglobulin-producing mouse myeloma and a nonproducing mouse lymphoma

A hybrid cell line was established by cell fusion between a cloned Balbic myeloma that is resistant to 8-azaguanine and produces immunoglobulin (gammaG and free kappa chain) and C57BL/6N lymphoma that is resistant to bromo-deoxyuridine and does not produce immunoglobulins. The hybrid cells contained the membrane antigens of both parents; they synthesized free kappa chain; no synthesis of gammaG (gamma(2a)) heavy chain was detected.