The RNA interference

The phenomenon of RNA interference (RNAi) is highly conserved mechanism in the organism evolution. As a immune system, RNAi is a ubiquitous mechanism against invading microorganism in plant and animal cells. Recently, it has been found that RNAi is the process by which double-strand RNA(dsRNA) directs sequence-specific degradation of messenger RNA and the mediations of sequence specific messenger RNA degradation are 21-and 23-nucleotide small interfering RNAs that generate by ribonuclease from endogenous longer dsRNA or by transfectious technics from heterologous dsRNA. Over the past few years, the way in which cells respond to dsRNA by silencing homologous genes has revealed a new regulating paradigm in biology.     

The physical appearance of the vulva in ureaplasma infected ewes

The vulvar labia of ewes naturally infected with ureaplasma were significantly more swollen and red than those of uninfected ewes. Similar changes were observed following experimental infection of previously uninfected ewes. These differences, although statistically significant, were not sufficiently marked to be useful in diagnosing infection by clinical examination.     

Individual antigens of cattle. Bovine CD2 (BoCD2).

The data obtained in the workshop provide further evidence that CH128A and IL-A26 and the 12 new mAbs that form a cluster recognise the bovine orthologue of CD2. The mAbs inhibit rosetting with SRBC, stain cells in primary and secondary lymphoid organs in patterns consistent with those obtained in humans with anti-CD2 mAbs, and the 11 IgG mAbs all immunoprecipitate a peptide with a Mr of 58-62 kDa. It is not clear from the studies whether the epitopes defined by the mAbs correspond with the region I and II epitopes present on CD2. None of the data suggest that any of the mAbs recognise the region III (CDD2R) epitope (Peterson and Seed, 1987; Knapp et al., 1989). Further studies are now needed to define the physical and functional relation of the epitopes and establish whether antibody-mediated activation corresponds with that noted in humans. Data reported in one study (Baldwin et al., 1988) with IL-A26 suggest possible differences in the requirements for activation. In addition, further studies are needed to demonstrate how many cell types express BoCD2. In mice, evidence has been presented which shows the mouse orthologue is expressed on some B cells (Yagitta et al., 1989). Studies in cattle have clearly shown CD2 is present on the majority of CD4+ and CD8+ T-cells and a small population of CD4-/CD8- cells (Baldwin et al., 1988; Davis, unpublished observations). Evidence presented in this workshop has shown that some CD2+ cells express a WC2 molecule (Sopp et al., 1991).(ABSTRACT TRUNCATED AT 250 WORDS)     

Multifactorial analysis of distance in studies of ecological community structure

A framework is described for partitioning the matrix of between-sample distances that forms a starting point for many ecological studies into contributions attributable to different factors in a structured data set. This partitioning enables a series of ordinations to be produced that better enable an insight to be gained into the effects of the factors. More detailed application of the same partitioning provides a decomposition of each factorial effect into single degree-of-freedom contrasts, which enables reasons for observed trends to be investigated. The methods are illustrated by application to data from a study of marine community structure.     

Culture of bovine bone marrow progenitor cells in vitro.

In vitro methylcellulose cultures of bovine bone marrow progenitor cells were developed. An existing technique described for bovine species was compared to a method for human tissue and further adapted during subsequent experiments. Bovine bone marrow samples were collected at the slaughterhouse, and mononuclear cells were separated by gradient centrifugation (1.077 g/ml specific density and 400 g). The use of 3% bovine leucocyte-conditioned medium, produced by stimulation of blood lymphocytes with 4 microg/ml concanavalin A and harvested on day 4 of culture, gave better results than the use of supernatant of the human bladder carcinoma 5637, which is widely used in human bone marrow cultures. However, bovine leucocyte-conditioned medium was not added to erythroid cultures because inhibitory effects were observed. Erythroid colonies were stimulated with erythropoietin, and hemin was added to enable microscopic identification. Reduced oxygen tension was necessary to induce growth of erythroid colonies. This was not necessary for myeloid cultures. In conclusion, the results of this study show that the growth of myeloid and erythroid colonies in methylcellulose-based medium requires different culture conditions, which are different from the culture conditions for human cells.     

Analysis of the reactivity of anti-bovine CD8 monoclonal antibodies with cloned T cell lines and mouse L-cells transfected with bovine CD8

Mouse L-cells transfected with bovine CD8 and two Theileria parva-infected cloned T cell lines expressing bovine CD8 were used to screen the panel of ten monoclonal antibodies (mAbs) submitted to the workshop. Eight of the ten mAbs reacted with the transfectant and both the cloned T cell lines. However, two mAbs CC58 and BAT82A did not recognise the transfectant and only reacted with one of the T cell lines. Further biochemical studies indicated that the eight mAbs react with both homo- and heterodimeric forms of bovine CD8 whilst the two mAbs CC58 and BAT82A react with only heterodimeric forms. These data suggest that bovine DC8 is encoded by two genes as is the case in mouse and man.     

Pharmacokinetics and pharmacodynamics of butorphanol in llamas after intravenous and intramuscular administration.

OBJECTIVE: To evaluate disposition of butorphanol after i.v. and i.m. administration, effects on physiologic variables, and analgesic efficacy after i.m. administration in llamas. DESIGN: Nonrandomized crossover study. ANIMALS: 6 healthy adult male llamas. PROCEDURE: Butorphanol (0.1 mg/kg [0.045 mg/lb] of body weight) was administered i.m. first and i.v. 1 month later. Blood samples were collected intermittently for 24 hours after administration. Plasma butorphanol versus time curves were subjected to pharmacokinetic analysis. Two months later, butorphanol (0.1 mg/kg) was administered i.m., and physiologic variables and analgesia were assessed. RESULTS: Extrapolated peak plasma concentrations after i.v. and i.m. administration were 94.8 +/- 53.1 and 34.3 +/- 11.6 ng/ml, respectively. Volume of distribution at steady state after i.v. administration was 0.822 +/- 0.329 L/kg per minute and systemic clearance was 0.050 +/- 0.014 L/kg per minute. Slope of the elimination phase was significantly different, and elimination half-life was significantly shorter after i.v. (15.9 +/- 9.1 minutes) versus i.m. (66.8 +/- 13.5 minutes) administration. Bioavailability was 110 +/- 49% after i.m. administration. Heart rate decreased and rectal temperature increased. Somatic analgesia was increased for various periods. Two llamas became transiently sedated, and 2 became transiently excited after butorphanol administration. CONCLUSIONS AND CLINICAL RELEVANCE: Although i.v. administration of butorphanol results in a short half-life that may limit its analgesic usefulness, the elimination half-life of butorphanol administered i.m. is likely to be clinically useful. The relationship among plasma butorphanol concentration, time, and analgesia differed with the somatic analgesia model; clinically useful analgesia may occur at lower plasma concentrations than those reported here.