Heat-induced stabilization of the nuclear matrix: A morphological and biochemical analysis in murine erythroleukemia cells

Using mouse erythroleukemia cells we performed a comprehensive morphological and biochemical study of the nuclear matrix obtained after exposure of isolated nuclei to 37 degrees C or from cells heat shocked in vivo at 43 or 45 degrees C. At the ultrastructural level it was possible to see that in the absence of a 37 degrees C incubation of purified nuclei, the final matrix lacked well-defined nucleolar remnants but a peripheral lamina was clearly visible, as well as a sparse fibrogranular network which was located at the periphery of the structures. On the contrary, after a 37 degrees C nuclear incubation, very electron-dense nucleolar remnants were observed along with an abundant meshwork dispersed throughout the interior of the structures. When intact cells were heat shocked in vivo, electron-dense residual nucleoli were present only when isolated nuclei had been exposed to 37 degrees C in vitro, whereas without such an incubation, they were not as easily distinguishable and appeared less electron-dense. In the latter case the inner network was more evenly distributed. After purified nuclei were incubated at 37 degrees C for 45 min, the high salt and DNase I resistant fraction retained about 18% of the nuclear protein whereas if the heating was omitted protein recovery dropped to 6%. An increase in the recovery of intact structures in the matrix fraction was the main reason for the higher protein recovery. Heating nuclei in vitro further increased the amount of nuclear protein present in the matrix fraction even if intact cells had been heat shocked in vivo. No major qualitative differences were seen when the polypeptide pattern of the various types of nuclear matrices was analyzed on one-dimensional polyacrylamide gels and this finding was further supported by Western blot analysis with a monoclonal antibody to lamins A and C. These results show that heating mainly stabilizes the nucleolar remnants of the matrix and to a lesser extent the inner network, but the morphology of the final structures is different depending on whether the stabilization is performed in vivo or in vitro.     

A T-cell antigen system of chickens: Ly-4 and Marek's disease

A search was made for lymphocyte antigens associated with resistance or susceptibility to the T-cell lymphoma induced by the herpes virus of Marek's disease (MD), the experimental model for Burkitt's lymphoma of humans. Antisera were produced by reciprocal immunization with whole blood between an MD-resistant and susceptible line of chickens compatible at the major histocompatibility complex (MHC), and were tested against lymphocytes of both lines. The lymphocytes were not agglutinated, immobilized, or lysed, but their ability to evoke graft-versus-host (GVH) splenomegaly was reduced. This inhibitory activity was line-specific, and these sera had a maximum limiting effect on GVH splenomegaly at a dilution of 1/50 and a minimum at 1/800 dilution. A test based on the differential limitation of GVH splenomegaly by a pair of alloantisera was used to identify the antigens in F₁ and F₂ generations. The segregation results established a locus,Ly-4, with two codominant alleles,Ly- 4^a andLy-4 ^b .Ly-4 is distinct from theA, B, orC blood group loci and from theBu-1 locus determining B-cell antigens, but may be linked to theTh-1 locus determining T-cell antigens (recombination frequency of 32 percent). Tentative evidence was obtained from comparisons of homozygous F₂ and F₃ progeny for association of theLy-4 allele characteristic of the susceptible line with increased incidence of MD.     

Comparison of vegetable shortening and cocoa butter as vehicles for cortisol manipulation in Salmo trutta

This study demonstrates that vegetable shortening and cocoa butter are two effective vehicles for intraperitoneal cortisol implants in juvenile teleosts, specifically brown trout Salmo trutta, residing in north temperate freshwater environments. Each vehicle showed a different pattern of cortisol elevation. Vegetable shortening was found to be a more suitable vehicle for long‐term cortisol elevation [elevated at 3, 6 and 9 days post treatment (dpt)], while cocoa butter may be better suited for short‐term cortisol elevation (only elevated at 3 dpt). Additionally, plasma cortisol levels were higher with cortisol–vegetable shortening than with cortisol–cocoa butter implants. Plasma glucose levels were elevated 6 and 9 dpt for fishes injected with cortisol–vegetable shortening, but did not change relative to controls and shams in cortisol–cocoa butter fishes. In conclusion, vegetable shortening and cocoa butter are both viable techniques for cortisol manipulation in fishes in temperate climates, providing researchers with different options depending on study objectives.     

Influence of non-MHC T lymphocyte alloantigens on regression of rous sarcomas in the chicken

Chickens of Regional Poultry Research Laboratory (RPRL) inbred line 63 regress sarcomas induced by Bryan high-titer Rous sarcoma virus to a greater extent than chickens of line 7₂, although these lines are identical for the major histocompatibility complex (MHC, B complex). They differ, however, at two independent autosomal loci, Ly-4 and Th-1, which determine surface alloantigens of partly overlapping subsets of T lymphocytes. Association of genotypes at these loci with quantitative variation in ability to regress Rous sarcomas was tested in segregating progeny derived from crosses of lines 6₃ and 7₂. In the F4 generation chickens of the Ly-4 ^a /Ly-4 ^a , Th-1 ^a /Th-1 ^a genotype (symbolized aa/aa) had significantly higher regressor ability than any of the other three double homozygous genotypes. In F5, all nine genotypes formed by combinations of homozygotes and heterozygotes were tested, and higher regressor ability was shown by the aa/aa, ab/aa, and aa/ab genotypes. These results indicate that higher regression is associated with: (1) interaction between the line 6₃ Ly-4 ^a and Th-1 ^a alleles in homozygous form; and (2) dominance x dominance interaction, in that the a allele at each locus is dominant for higher regression only within the homozygous aa genotype at the other locus.     

The effect of high hydrostatic pressure on Listeria monocytogenes in phosphate-buffered saline and model food systems

Three strains of Listeria monocytogenes (NCTC 11994, a poultry isolate and the Scott A strain) were exposed to a range of pressures (300, 350, 375, 400 and 450 MPa) in 10 mmol l⁻¹ phosphate-buffered saline (PBS) at pH 7·0 for up to 30 min at ambient temperature. Generally, increasing the magnitude and duration of compression resulted in increasing levels of inactivation, although the inactivation kinetics varied depending on the strain and pressure applied. The three strains also exhibited a wide variation in their resistance to high pressure. The resistance of the three strains to high pressure (375 MPa) was also assessed in a series of model food systems containing one of each of the three main food constituents: protein (1, 2, 5 and 8% w/v bovine serum albumin in PBS), carbohydrate (1, 2, 5 and 10% w/v glucose in PBS) and lipid (olive oil (30% v/v) in PBS emulsion). Overall, increasing the concentrations of bovine serum albumin (BSA) and glucose in the suspending medium resulted in decreasing levels of inactivation of all three strains; however, the minimum concentration of BSA and glucose required to increase survival to a level greater than that observed in PBS alone varied depending on the strain and on the duration of the treatment. The survival of all three strains was greater in the olive oil/PBS emulsion than in PBS alone at all treatment times.     

Incidence of Listeria spp. and Listeria monocytogenes in a Poultry Processing Environment and in Poultry Products and Their Rapid Confirmation by Multiplex PCR

Volume 60, no. 12, p. 4602: Fig. 1 should appear as shown below. FIG. 1. Incidence of L. monocytogenes and other Listeria spp. in a poultry processing environment and in raw and cooked poultry products from March to September 1992. (A) Raw (R) and cooked (C) areas within the processing environment. (B) Raw and cooked poultry products. (box), L. monocytogenes; (box), Listeria spp. (not L. monocytogenes); (symbl), no Listeria spp. [This corrects the article on p. 4600 in vol. 60.].     

Chemical and carbon isotopic composition of fatty acids in adipose tissue as indicators of dietary history in wild arctic foxes (A lopex lagopus) on Svalbard

The chemical and carbon isotope compositions of triacylglycerol fatty acids were analysed in samples from two or more adipose depots dissected from adult and subadult arctic foxes collected between November 1991 and March 1992 in four different areas of Spitsbergen in the Svalbard archipelago (latitude 78° 5' to 79° 50' N). Site-specific differences were minor but there were large and consistent differences in the fatty acid composition of the storage lipids of foxes caught in the same areas, suggesting that residents of contiguous territories may have had quite different diets. The adipose tissue of adult foxes caught in Austfjordneset, an area where reindeer are rare, contained a much greater proportion of unsaturated fatty acids, suggesting that these animals were feeding mainly in the marine ecosystem, probably on seabirds and/or fish in summer and the remains of polar bear kills in winter. Measurements of the relative abundance of the carbon isotopes ¹²C and ¹³C in individual fatty acids show that palmitoleic acid (C16:1) in storage triacylglycerols originates from the marine food chain, probably together with most other unsaturated lipids, but that the foxes obtain oleic acid (C18:1), and probably most saturated fatty acids, from either terrestrial or marine sources.