Bovine viral diarrhea virus is inactivated when whole milk from persistently infected cows is heated to prepare semen extender.

Bovine viral diarrhea virus (BVDV) can be present in cryopreserved bovine semen and be transmitted through artificial insemination. Because BVDV can be shed in milk, the virus might also be introduced as a contaminant of milk-based semen extenders. Thus, the purpose of this study was to evaluate the epidemiologic risk of using heated, BVDV-contaminated milk to prepare semen extender. Milk was obtained from cows free of and persistently infected (PI) with BVDV. Six replicates of milk samples were processed by heating (85-92.2 degrees C, 10min). Samples of milk collected before and after heating were assayed for BVDV. Additionally, milk was injected intravenously into eight BVDV seronegative calves to monitor for seroconversion and viral infection. Virus was not detected in any milk samples from negative animals. Virus was consistently isolated from unheated milk samples from PI cows by passage of somatic cells, ultracentrifugation, and animal inoculation. Virus was usually detected in these samples by RT-nPCR (reverse transcription nested polymerase chain reaction). In heated milk samples from PI cows, no infectious BVDV was detected using any technique, but viral RNA was detected using RT-nPCR in four of six replicates. Bovine viral diarrhea virus in milk from PI cows was inactivated by heating. Therefore, properly heated milk used in semen extenders will not result in transmission of infectious BVDV. Although RT-nPCR detected the presence of viral RNA in milk samples after heating, the virus was not infectious as demonstrated by lack of replication despite using multiple sensitive techniques.

Identification of mRNA species that code for extra-embryonic endodermal cytoskeletal proteins in differentiated derivatives of murine embryonal carcinoma cells.

The messenger RNA species that code for two extra-embryonic endodermal cytoskeletal proteins (Endo A and Endo B) have been identified. Endo A and Endo B messenger RNA species are differentially expressed. They are absent, or at basal levels, in undifferentiated embryonal carcinoma cells (F9.22 cell line) and relatively abundant in parietal endoderm (PFHR9 cell line) or in embryonal carcinoma cells that have been induced, by retinoid acid, to differentiate to parietal endoderm. Endo A and Endo B messenger RNA can be detected in F9.22 cells 48 to 72 h after exposure to retinoic acid, which is coincident with the expression of Endo A and Endo B proteins. The size of Endo A and Endo B messenger RNA has been determined by denaturing methyl mercury hydroxide agarose gels to be 2.0 +/- 0.1 and 1.5 +/- 0.2 kilobases, respectively.

Construction and identification of bacterial plasmids containing nucleotide sequence for human leukocyte interferon.

Bacterial plasmids containing human leukocyte interferon sequences were constructed and identified. Identification was confirmed by correspondence of the nucleotide sequence with out amino acid sequence of human leukocyte interferon. The finding of bacterial recombinants containing distinct leukocyte interferon sequences is consistent with our purification of different leukocyte interferon species. We conclude that what has been designated human leukocyte interferon is, indeed, a class of homologous proteins. Preliminary indications suggest that their diversity appears to be represented by individual genomic equivalents. Each of the individual species exhibits characteristic activities. The structural modulation of these biological activities has immense significance for understanding the natural role of the interferons and for refining and developing their ultimate therapeutic potential.

Human leukocyte interferon produced by E. coli is biologically active.

A human leukocyte interferon cDNA was enzymatically synthesized, inserted into the vector pBR322, and cloned in Escherichia coli. The DNA sequence codes for a 23-amino acid signal peptide followed by an interferon polypeptide of 165 amino acids. An expression plasmid was constructed which permits the synthesis in E. coli of 2.5 x 10(8) units of interferon per litre of culture. This LeIF protected squirrel monkeys from lethal encephalomyocarditis virus infection.

A coliphage lambda vector with enhanced biological containment: lambda gtALO.lambda B.

The biological containment of the lambda gt family of cloning vectors has been enhanced by conditionally blocking DNA replication as well as head and tail morphogenesis. The vector, lambda gtALO.lambda B, was constructed by crossing the Oam29, Aama1 and Lam439 mutations into lambda gt.lambda B. The mutation blocking phage DNA replication, Oam29, is suppressed by suII+ or suIII+. The head gene mutation, Aama1, is suppressed by suIII+ but not by suII+ and the tail gene mutation, Lam439, is suppressed by suII+ but not by suIII+. This allows the option of increasing the biological containment by producing heads when a large amount of cloned DNA is being prepared from an individual isolate. A model recombinant, lambda gt Aama1 Lam439 Oam29.KmR' (lambda gtALO.KmR') was constructed and the containment of the vector was evaluated by the series of standardized experiments required for EK2 certification.

Somatic, behavioral, and reproductive disturbances in mice following neonatal administration of sodium L-aspartate.

Sodium L-aspartate (ASP) was administered to neonatal mice according to an increasing dose schedule from Days 2--11 after birth. Adult ASP-treated animals showed large increases in body weight over controls along with stunted body length. The ASP group also showed decreases in locomotor and exploratory behavior. Reproductive dysfunction occurred in both female and male ASP-treated animals. Among treated animals, females had fewer pregnancies and smaller litters while males showed reduced fertility. Evidence of multiple endocrine dysfunction in ASP-treated animals was reflected by decreased pituitary, thyroid, ovaries and tested weights, along with delayed onset of puberty in females. These results demonstrate that sodium L-aspartate produces a syndrome similar to that seen following the administration of monosodium L-glutamate.