Inhibition of growth of cultured Babesia microti by serum and macrophages in the presence or absence of T cells

Serum and macrophages from the acute-phase (days 12-14 p.i.) and recovery-phase (days 23-25 p.i.) of infection of mice with Babesia microti were analyzed for their ability to inhibit the in vitro growth of B. microti in the presence or absence of T cells. Recovery-phase serum was inhibitory to the growth of B. microti, whereas, acute-phase serum had no inhibitory effects. Both acute- and recovery-phase macrophages inhibited B. microti growth. The co-culture of acute- but not recovery-phase T cells with macrophages from uninfected control mice was inhibitory to the growth of B. microti. Growth of B. microti was also inhibited in cultures containing macrophages from uninfected control mice plus culture supernatant fluid from acute-phase but not recovery-phase T cells. The supernatant fluid from B. microti cultures with acute-phase T cells contained IFN-gamma detected by a sandwich ELISA, whereas cultures with control T cells or recovery phase T cells did not. Results of the present study suggest the likelihood of a protective role against B. microti in mice for antibody which appeared in recovery-phase serum and for macrophages activated by IFN-gamma from acute-phase T cells.     

Effects of Mild and Moderate Monoclonal Antibody Dose on Inflammation,Bone Loss,and Activation of the Central Nervous System in a Female Collagen Antibody-induced Arthritis Mouse Model

Induction of severe inflammatory arthritis in the collagen antibody-induced arthritis (CAIA) murine model causes extensive joint damage and pain-like behavior compromising analysis. While mild models are less severe, their reduced, variable penetrance makes assessment of treatment efficacy difficult. This study aimed to compare macroscopic and microscopic changes in the paws, along with central nervous system activation between a mild and moderate CAIA model. Balb/c mice (n=18) were allocated to control, mild, and moderate CAIA groups. Paw inflammation, bone volume (BV), and paw volume (PV) were assessed. Histologically, the front paws were assessed for joint inflammation, cartilage damage, and pre/osteoclast-like cells and the lumbar spinal cord and the periaqueductal gray (PAG) region of the brain for glial reactivity. A moderate CAIA dose induced (1) significantly greater local paw inflammation, inflammatory cell infiltration, and PV; (2) significantly more osteoclast-like cells on the bone surface and within the surrounding soft tissue; and (3) significantly greater glial reactivity within the PAG compared with the mild CAIA model. These findings support the use of a moderate CAIA model (higher dose of monoclonal antibodies with low-dose lipopolysaccharide) to induce more consistent histopathological features, without excessive joint destruction.     

Frameshift mutagenesis of lambda prophage by 9-aminoacridine, proflavin and ICR-191

Summary The changes in DNA base sequence induced in the lambda cI gene in an E. coli lysogen have been determined following mutagenesis by three acridine derivatives: 9-aminoacridine and proflavin, which bind reversibly to DNA; and ICR-191, which attaches covalently to DNA through a half-mustard group. For all three derivatives, most mutations are +1 and-1 frameshifts in runs of adjacent G:C pairs. The specificity of mutagenesis at various sites is similar for all three compounds. Prophage in mutL host cells, deficient in mismatch repair, are much more susceptible to mutagenesis by 9-aminoacridine. The induced mutations are also frameshifts, and the site specificity is the same as in lysogens of wild type cells. Thus, additions or deletions of single bases can be corrected by the mismatch repair system, but mismatch repair does not play an important role in determining the sequence specificity of the mutational events.     

Changes in DNA base sequence induced by targeted mutagenesis of lambda phage by ultraviolet light

In targeted mutagenesis of lambda phage by ultraviolet light, the mutations are caused by radiation-induced lesions in the phage DNA. Of 62 mutations in the lambda cI gene that were sequenced, 41 (63%) of the targeted mutations were transitions, with similar numbers of C X G to T X A and T X A to C X G base changes. The remaining 21 mutations were about equally divided among eight transversions, seven frameshifts (5 additions and 2 deletions), and six double events with either two nearby base changes or a base change and a nearby frameshift. Of the 62 mutations, 60 could be associated with -Pyr-Pyr- sequences in the DNA, sites of likely photoproducts. For more information on this point, lambda phage were irradiated with 313 nm light in the presence of acetophenone, for which the major photoproduct is reported to be the thymine-thymine cyclobutyl dimer, with no measurable Pyr(6-4)Pyo photoproducts. Of 22 mutations sequenced, 19 were transversions and only one was a transition, permitting the conclusion that thymine-thymine cyclobutyl dimers are not the primary cause of ultraviolet light-induced transitions. A consideration of all the data strongly suggests that Pyr(6-4)Pyo photoproducts are mutagenic lesions.     

Emergence and development of resistance to antimicrobial chemicals and heat in spores of Bacillus subtilis

The emergence and development of chemical and thermal resistance in spores of Bacillus subtilis was examined. The chemicals studied were of the disinfectant type: glutaraldehyde, hypochlorite, hypochlorite-methanol and povidone-iodine. Growth and sporulation were followed by electron microscopy and resistance assigned to specific stages in relation to 45Ca and DPA accumulation. A sequential development of resistance was observed with thermal resistance appearing first at early Stage V corresponding to maturation of cortex and deposition of rudimentary spore coat material. Chemical resistance coincided with middle to late Stage V dependent on the chemical concerned. A progressive development of resistance was observed on prolonged incubation in sporulation medium and was affected by inclusion of lysozyme in the spore washing sequence.     

Lectin-mediated targeting of liposomes to a model surface. An ELISA method

Wheat germ agglutinin has been conjugated to the surfaces of sonicated phospholipid liposomes by reacting the protein derivatised with N-succinimidyl-S-acetylthioacetate (SATA) with the m-maleimidobenzoyl-N-hydroxysuccinimide (MBS) derivative of dipalmitoylphosphatidylethanolamine (DPPE) incorporated into the liposomal bilayers. The liposomes as characterised by photon correlation spectroscopy had a weight-average radius of 44 +/- 10 nm and the number of WGA molecules per liposome was in the range up to approx. 120. An ELISA method has been developed to assess the efficiency of targeting the conjugated liposomes to the antigenic determinants on a surface coated with glycophorin A (blood group B). For liposomes in which the degree of conjugation was controlled by varying the mol% DPPE-MBS from 3 to 27% targeting efficiency as assessed from the extent of inhibition of the ELISA increased by a factor of 10.