Detection and characterization of simian retroviruses homologous to human T-cell leukemia virus type I

Lymphoid cell lines were established from five different species of monkeys which were positive in antibodies cross-reactive with human T-cell leukemia virus type I (HTLV-I) and were shown to contain provirus sequences homologous to HTLV-I. Gene-specific probes of HTLV-I, gag, pol, env, pX, and LTR, hybridized efficiently with monkey DNAs from these cell lines under stringent conditions, indicating that the proviruses are very similar to HTLV-I along with whole viral genomes. However, the preliminary restriction mapping turned out the difference between simian retroviruses and HTLV-I and also among simian retroviruses. These findings suggest a common ancestor of simian and human retroviruses, but exclude the recent interspecies transmission between monkeys and humans.     

Selection of a mutant S1 gene during reovirus persistent infection of L cells: role in maintenance of the persistent state

LR-7 cells, variant L cells derived from a type 3 reovirus persistently infected (p.i.) carrier culture (R. Ahmed, W. M. Canning, R. S. Kauffman, A. H. Sharpe, J. V. Hallum, and B. N. Fields, Cell 25, 325-332, 1983) were used to define the viral genes critical for maintenance of the persistent state. A cloned viral isolate (L/C virus) derived from the p.i. culture replicated normally in LR-7 cells, while wild-type (wt) viruses of the three reovirus serotypes replicated less efficiently. To identify the viral gene(s) permitting enhanced replication of L/C virus in LR-7 cells, viral reassortants were prepared by mixed infection of L cells with L/C virus and type 1 wt. Study of the one-step growth curves and final yields of large numbers of reassortants in both L cells and LR-7 cells revealed that the presence of the S1 gene from L/C virus was critical for normal viral replication in LR-7 cells. However, this phenotype was suppressed by the simultaneous presence in reassortants of both the M2 and S4 genes from the type 1 wt parent. The critical change in the S1 gene occurred by passage 13 (63 days) after initiation of the carrier culture. Although multiple mutations are present in the viral population from p.i. cultures, certain specific mutations can be identified as critical for maintenance of the persistent state.     

Transcription and replication of eight RNA segments of influenza virus

A novel quantitation system of both plus- and minus-strand RNAs for all eight genome segments of influenza virus was developed using single-strand cDNAs as the probes for hybridization, and employed for the measurement of various RNA species in influenza virus WSN-infected MDBK cells. The synthesis rate and accumulation level of plus-strand RNAs differed considerably among eight RNA segments and were under temporal control. In contrast, eight vRNA molecules of minus polarity were synthesized coordinately at similar rates. Newly synthesized plus-strand RNAs were rapidly transported into the cytoplasm, particularly during the early phase of virus infection, but vRNAs accumulated in the nuclei until the late infection phase. The present data supported the differential regulation of synthesis and the separate transport between plus- and minus-strand RNAs.     

Genome locations of temperature-sensitive mutants in glycoprotein gB of herpes simplex virus type 1

A plasmid containing a herpes simplex virus type 1 (HSV-1) insert from strain KOS, prototypic coordinates 0.345 to 0.368 (3.45 kilobases) was mutagenized in vitro, and potential mutations were introduced into intact viral DNA by cotransfection. Functions normally associated with the glycoprotein gB are in the 1-9 complementation group, and the above coordinates include those that specify the gB glycoprotein gene. Following cotransfection, individual plaques were screened for temperature sensitivity (ts) of viral growth. A total of seven ts mutants was obtained, of which four were spurious mutations due to alterations outside the cloned sequences, presumably mediated by some aspect of the Ca-precipitation-cotransfection method. The remaining three did not complement known mutants of the 1-9 complementation group. These three mutants, along with tsJ12 (P.A. Schaffer, G.M. Aron, N. Biswal, and M. Benyesh-Melnick, 1973, Virology 52, 57-71) and tsJ33 (C.-T. Chu, D.S. Parris, R.A.F. Dixon, F.E. Farber, and P.A. Schaffer, 1979, Virology 98, 168-181), were physically located by marker-rescue experiments to three different restriction fragments between 0.345 to 0.368 map units. Sodium dodecyl sulfate-gel electrophoresis was used to analyze the glycoproteins synthesized during continuous or pulse-chase labeling protocols. All five mutants were found to synthesize a precursor of gB but did not accumulate mature gB during a pulse, a chase, or continuous labeling at the nonpermissive temperature.     

Effect of antibody-induced modulation of measles (SSPE) virus membrane proteins on beta-adrenergic receptor-mediated adenylate cyclase activity

The effect of measles virus antiserum on the expression of viral glycoproteins on the membranes of measles (SSPE) persistently infected C6 rat glioma cells was studied. There was a gradual loss of virus membrane antigen from these cells until no antigen was detectable 18 days after initiation of antiserum treatment. At this stage approximately 25% of cells still displayed intracellular virus antigen which was also lost after further cell passages. There was an accompanying recovery of the previously reported disrupted catecholamine-dependent beta-adrenergic receptor-stimulated cAMP synthesis in antiserum-treated cells which coincided with the loss of viral antigen from the membrane. This was determined to be due to a recovery of fluoride-stimulated activity of the cAMP synthesising adenylate cyclase enzyme to normal values. Thus we demonstrate that the impairment of this important cell function was due to insertion of viral antigen in the cell membrane rather than its accumulation in the cytoplasm.     

The fidelity of phosphorylation of the adenovirus DNA-binding protein by an in vitro nuclear protein kinase from virus-infected cells.

The type 5 adenovirus single-strand-specific DNA-binding protein is phosphorylated and can be labeled by the addition of ³²PO₄ to the medium of infected cells. An infected cell nuclear protein kinase activity is capable of phosphorylatiog the single-strand-specific DNA-binding protein in vitro (A. D. Levinson, E. H. Postel, and A. J. Levine. The fidelity of phosphorylation by this in vitro protein kinase activity was tested by comparing the ³²P-labeled partial peptides generated by Staphylococcus aureus protease digestion of the ³²P-labeled DNA-binding protein which had been phosphorylated in vivo or in vitro. Identical ³²P-labeled partial peptides were obtained from the DNA-binding protein whether it was phosphorylated in vivo or in vitro. A second nuclear protein of 36,000 MW was also faithfully phosphorylated by the in vitro nuclear protein kinase activity.     

Production and characterization of antisera specific for the erb-portion of p75, the presumptive transforming protein of avian erythroblastosis virus

The polypeptide pattern of intracytoplasmic A particles, isolated from mammary tumors of ICRC mice, displayed a prominant doublet of about 75,000 molecular weight (Ap75) and a lesser band of 42,000 molecular weight (Ap42). A particle proteins were analyzed by tryptic fingerprint mapping and compared to similarly treated core proteins (p10, p14, p23, and p28) of murine mammary tumor virus (MuMTV) purified from the milk of ICRC mice. Comparison of the peptide maps showed that almost all the major spots of the MuMTV core proteins could be located within the map of Ap75, and that Ap42 contained peptides of only p28 and p14. Since tryptic peptide maps of MuMTV core proteins have been shown to be strain-specific markers, we conclude that the A particles present in mammary tumors of ICRC mice are coded for by the same genes that code for the virions present in the milk of these mice.     

Mediated smoking cessation programs in the Stanford five-city project

Two mediated smoking cessation programs were subjected to a field evaluation. The Quit Kit is a printed self-help package, and "Calling It Quits" consists of five segments which were aired on the local television news. A sample of 239 persons requested the Quit Kit and were followed. At the 2-month and 12-month follow-ups, respectively, 13.6% and 17.9% of those surveyed reported abstinence. Results indicate the potential of mediated interventions.     

Selective and nonselective shunts for variceal bleeding. A prospective study of 103 patients

Based on the experience reported herein, the following conclusions have been made: (1) Although nonoperative means, including sclerotherapy, have an important role in the management of bleeding varices, they are not definitive means of treating recurrent variceal hemorrhage. (2) Because of the maintenance of hepatopetal flow and splanchnic venous hypertension, a selective shunt is associated with a lower incidence of encephalopathy and provides a better quality of life than does a nonselective shunt. Thus, an elective distal splenorenal shunt is the elective operation of choice for recurrent variceal hemorrhage. (3) Nonselective shunts can be performed with similar expectation of patient survival as selective shunts, but because of increased encephalopathy, should be reserved for emergency operations, in cases of unsuitable venous anatomy, and in those patients with intractable ascites. (4) A well-conceived elective shunt procedure can be performed with low operative mortality and long-term patency, results in significant survival, and is still considered the "gold standard" for treatment of variceal bleeding.     

Ejaculatory incompetence: a theoretical formulation and case illustration

Three theoretical formulations of ejaculatory incompetence have been proposed in the literature. They include: (1) aversive conditioned inhibition of the ejaculatory reflex, (2) an "autosexual" orientation associated with discrepant levels in the cognitive and physiological dimensions of sexual arousal, and (3) a discriminative learning model. These three models are discussed in relation to their theoretical and therapeutic implications. Clinical data supporting the discriminative view is presented.