Search

The genome of the class II filamentous bacteriophage Pf1 has been sequenced by a combination of the chain termination and chemical degradation methods. It consists of 7349 nucleotides in a closed, circular loop of single-stranded DNA. The size and position of its open reading frames (ORFs) in general resemble those of other filamentous bacteriophage genomes. The size and position of the spaces between the ORFs have not been conserved, however, and six short reading frames (2 of which overlap) occupy a region corresponding to that filled by genes 2 and 10 in the Ff genome. Most of the ORFs are preceded by sequences resembling ribosome binding sites from the phage's host. Pseudomonas aeruginosa, that appear to differ somewhat from their counterparts in Escherichia coli. A search for sequences related to known pseudomonad promoters suggests that the promoters in this bacteriophage may well be ntr-dependent, with the two strongest preceding the gene for the major coat protein (gene 8) and another ORF (430). Gene 8 is followed by a sequence with the properties of a rho-independent terminator of transcription, like that at the same position in the genome of Ff. The Pf1 genome contains no collection of potential stem-and-loop structures corresponding to those that initiate replication of Ff DNA and assembly of the Ff virion, although isolated structures of this kind are present. The available evidence suggests that at least 13 of the 14 major ORFs are expressed. Overall, the organization of the Pf1 genome differs from that of the other class II filamentous phage whose genome has been sequenced, Pf3, as much as it does from that of the class I phages Ff and IKe.

Developing and nurturing professional relationships with support departments enhance the function of any nursing area. Building mutual goals and sharing common knowledge of patient cases can create improved patient care in the endoscopy suite and the inpatient unit. This article discusses possible methods to achieve team spirit and a sense of common purpose for the gastrointestinal nurses and associates who share a client in the procedure area and the ward.

The amino-acid sequence of the single-stranded DNA-binding protein of bacteriophage Pf1 and the nucleotide sequence of the corresponding gene have been determined. The protein has 144 amino acids and a molecular weight of 15 400; the gene consists of 435 nucleotides. The amino-acid sequence was determined by Edman degradation, carboxypeptidase A, B, and P digestion of intact protein and of peptides derived by chymotrypsin, Staphylococcus aureus V8 protease, and trypsin digestion. The nucleotide sequence was determined by the dideoxy method after random cloning of fragments of Pf1 DNA into M13. No sequence homology could be established between the amino-acid sequence of the DNA-binding protein of Pseudomonas aeruginosa-specific bacteriophage Pf1 and bacteriophage fd of Escherichia coli.

To determine development rates, fecundity and survival of Rhipicephalus appendiculatus, Boophilus decoloratus and B. microplus, a study was carried out in long and short grass in the highveld of Zimbabwe. Engorged adult females of the three species and engorged larvae and nymphs of R. appendiculatus were buried beneath the soil in small cages in the rainy, cool and hot seasons in 1980 and 1981. Half the number of cages were examined regularly to determine development rates and half were left undisturbed to determine survival rates and the fecundity of engorged females. Development was most rapid during warm conditions and slowest during cool conditions, but high temperatures appeared to prolong the preoviposition periods of all species. The relationship between fluctuating temperatures and rate of development in the field was defined using a least-squares procedure. Survival of engorged females was usually high, but was reduced by predation when they were not protected. Fecundity was reduced in long grass during the cool season and in short grass during the hot season. A higher percentage of eggs hatched in the rainy season than in the cool or dry seasons. The survival of engorged larvae and nymphs was usually high in all seasons. Engorged nymphs were the hardiest stage and eggs the most suceptible stage to adverse microclimatic conditions.

Bunyamwera virus replication was examined in Aedes albopictus (mosquito) cell cultures in which a persistent infection is established and in cytopathically infected BHK cells. During primary infection of A. albopictus cells, Bunyamwera virus reached relatively high titers ( approximately 10(7) PFU/ml), and autointerference was not observed. Three virus-specific RNAs (L, M, and S) and two virion proteins (N and G1) were detected in infected cells. Maximum rates of viral RNA synthesis and viral protein synthesis were extremely low, corresponding to <2% of the synthetic capacities of uninfected control cells. Viral protein synthesis was maximal at 12 h postinfection and was shut down to barely detectable levels at 24 h postinfection. Virus-specific RNA and nucleocapsid syntheses showed similar patterns of change, but later in infection. The proportions of cells able to release a single PFU at 3, 6, and 54 days postinfection were 100, 50, and 1.5%, respectively. Titers fell to 10(3) to 10(5) PFU/ml in carrier cultures. Persistently infected cultures were resistant to superinfection with homologous virus but not with heterologous virus. No changes in host cell protein synthesis or other cytopathic effects were observed at any stage of infection. Small-plaque variants of Bunyamwera virus appeared at approximately 7 days postinfection and increased gradually until they were 75 to 95% of the total infectious virus at 66 days postinfection. Temperature-sensitive mutants appeared between 23 and 49 days postinfection. No antiviral activity similar to that reported in A. albopictus cell cultures persistently infected with Sindbis virus (R. Riedel and D. T. Brown, J. Virol. 29: 51-60, 1979) was detected in culture fluids by 3 months after infection. Bunyamwera virus replicated more rapidly in BHK cells than in mosquito cells but reached lower titers. Autointerference occurred at multiplicities of infection of approximately 10. Virus-specific RNA and protein syntheses were at least 20% of the levels in uninfected control cells. Host cell protein synthesis was completely shut down, and nucleocapsid protein accumulated until it was 4% of the total cell protein. We discuss these results in relation to possible mechanisms involved in determining the outcome of arbovirus infection of vertebrate and mosquito cells.

This article describes 10 special therapeutic tubes utilized in gastroenterology. The discussion includes a description of each tube; categories are developed by anatomical site as well as the number of lumens in each tube. Several tubes are not discussed due to previous development in the literature or specialty purposes limited to diagnostics: esophageal manometry, Levin, Salem sump, gastrostomy tubes, bougies, dilators, the Dreiling tube and the Rubin-Quinton tube. Many of the tubes discussed are often utilized in the inpatient setting on a ward and may not be as familiar to the gastroenterology nurse or associate in an endoscopy suite.

We have characterized members of the Mapputta group of 'bunyavirus-like' viruses in terms of morphology, structure, ultrastructural development and virus-directed RNA and protein synthesis. Our primary study has been with Maprik virus (MPK) as a representative of the group. The MPK virion is uniformly spherical (congruent to 90 nm diameter) and possesses a membrane envelope. Virus maturation is by budding into small vesicles in the perinuclear region. During infection of BHK cells which is cytopathic, maximum viral RNA synthesis is congruent to 20% of RNA synthesis in uninfected cells without actinomycin D. In Aedes albopictus cells, where high initial titres are followed by a persistent infection, maximum viral RNA synthesis is only 1.5% of levels in control cells. MPK virions contain three RNA species. MPK generates four virus-specific RNAs (L, M, S1 and S2) and three nucleocapsid species in infected BHK cells. Six virus-specific polypeptides are present in infected cells; the four largest correspond to L (a probable transcriptase component), G1 and G2 (envelope proteins) and N (the nucleocapsid protein), respectively. These results and complementary studies with Trubanaman and Gan Gan viruses indicate that the members of the Mapputta serogroup can be placed in the bunyavirus genus of the family Bunyaviridae.

Adenylate cyclase activity of a rat embryo fibroblast cell line (F111) is markedly increased by brief treatment with 1:300 trypsin. The degree of stimulation depends upon the length of time the cells are treated and the concentration of trypsin. Crystalline trypsin produced a stimulation similar to that obtained with 1:300 trypsin. Further, the addition of soybean trypsin inhibitor blocked the stimulation of adenylate cyclase by 1:300 trypsin. Trypsin-treated adenylate cyclase responds to PGE1, but there is no increase over that of untreated enzyme. This result and the increase in fluoride-stimulated levels of activity suggest that the trypsin is acting upon the catalytic unit of the enzyme.

Rhipicephalus appendiculatus passes through one generation per annum in the highveld of Zimbabwe, Rhodesia and shows a well-defined pattern of seasonal activity. Peak adult activity occurs in the rainy season, whereas peak larval and nymphal activity occurs in the dry season. Adult activity is regulated by the combined influences of temperature, humidity, and day length. Climatic factors have little or no direct influence on the activity of larvae and nymphs. The occurrence of the larval and nymphal activity peaks is determined by the timing of the adult activity peak and the duration of the preceeding developmental periods, which are temperature dependent. In the early rainy season, unfed adults climb to the tips of the grass and enter a period of quiescence prior to becoming active.

Barmah Forest virus has been characterized in a number of ways including electron microscopy of infected cells; physical studies of the virion, its RNA, and associated proteins; N-terminal sequence analysis of the two envelope glycoproteins; studies of macromolecular species present in infected cells; and serological cross-reactions with alphaviruses and bunyaviruses. From these results Barmah Forest virus is clearly an alphavirus since the structure of the virion, the mode of replication, and the macromolecular species present in infected cells are typical of alphaviruses. The N-terminal regions of the two glycoproteins E1 and E2 show extensive sequence homology (approximately 50%) with those of other alphaviruses. Barmah Forest virus cross-reacts in hemagglutination inhibition tests, although not in complement fixation tests or infectivity neutralization tests, with other alphaviruses. In some of its properties Barmah Forest virus is unusual, however. It cross-reacts in complement fixation and hemagglutination inhibition tests with Umbre virus, a bunyavirus, which originally led it to be classified as a bunyavirus; the glycosylation pattern of E2 of Barmah Forest virus appears to differ from that of other alphaviruses; and the sedimentation coefficient of the virion appears to be slightly less than that of other alphaviruses.