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The relation between polyoma T-antigen and increased 5S RNA synthesis in cell-free extracts from polyoma-infected mouse kidney cell cultures.

In polyoma-infected mouse kidney cell cultures 5S RNA synthesis began to increase around 16 h, i.e. 7-9 h after the onset of polyoma T-antigen synthesis. The rate of polyoma-induced 5S RNA synthesis reached a maximum plateau around 25 h when it was 1.8-2.0 times higher than in mock-infected parallel cultures. Stimulation of 5S RNA synthesis in vivo thus coincided in time with the increase in total cellular RNA and protein. Cell-free extracts (S100) prepared at 15 h from mock-(S100-M) or polyoma-infected (S100-Py) mouse kidney cell cultures were indistinguishable with respect to protein concentration and 5S RNA synthesis, using a cloned somatic Xenopus borealis 5S gene as template. S100-Py extracted 25 h after infection contained 30% more protein and synthesized 1.5-2.0 times more 5S RNA than S100-M. Complete removal of the polyoma T-antigens from S100-Py by 3 cycles of immunoprecipitation with hamster anti-T serum remained without effect on stimulated 5S RNA synthesis. However, a linear relationship between 5S RNA synthesis and protein concentration of S100-M and S100-Py was observed.

Animals↗

Nematospiroides dubius and Nippostrongylus brasiliensis: delayed type hypersensitivity responses to ovalbumin in the infected mouse.

Mice (C57BL) infected with the intestinal nematode Nematospiroides dubius showed depressed delayed type hypersensitivity responses to ovalbumin administered subcutaneously in Freund's complete adjuvant. IgG and IgM responses to this inoculum were unaffected. It is unlikely that the depression arose from impairment of the ear test response because responses to an extract of the adult parasite were measurable and ear testing with lipopolysaccharide yielded normal responses in infected mice. Furthermore, mice immunized on the day of infection responded normally, whilst long term infected mice ear challenged with antigen pulsed macrophages gave depressed responses. The in vitro proliferative responses of cells from the spleens and from the lymph nodes draining the site of immunization were enhanced marginally by N. dubius infection. Furthermore, these cells induced normal or elevated adoptive delayed-type hypersensitivity and IgG responses in irradiated recipients. These findings suggest that N. dubius does not compromise the development of ovalbumin specific T cells involved in a delayed type hypersensitivity response. Evidence for the induction of suppressor cells by N. dubius is discussed, and the findings are compared with results obtained with Nippostrongylus brasiliensis, a parasite which is rejected rapidly from the mouse.

Animals↗

The relationship of phosphorylation of membrane proteins with the osmotic fragility and filterability of Plasmodium berghei-infected mouse erythrocytes.

Membrane from Plasmodium berghei-infected mouse erythrocytes showed a pattern of protein phosphorylation which was substantially altered from the normal pattern, with an increase in the phosphorylation of the protein with an apparent molecular weight of 43,000 (M 43), which increased from undetectable in uninfected cells to a maximum in the mature trophozoite stage. Phosphorylation levels of this and other minor bands were strongly correlated with osmotic fragility and filterability. The level of M 43 phosphorylation in membranes from cells which remained intact in a hypotonic medium was 3.82 +/- 0.59-times that of lysed cells, compared with the value of 0.76 +/- 0.07 calculated from distribution alone. Results found when intact erythrocytes were phosphorylated by incubation with [32P]Pi prior to partial lysis were similar to those found when membranes from the lysed and unlysed fractions were subsequently phosphorylated with [gamma-32P]ATP. Infected erythrocytes which could pass repeatedly through 3-micron polycarbonate filters had a much higher phosphorylation level for the M 43 region than whole infected cells with similar parasitemia and stage distribution. The phosphorylation change could play a role in the control of osmotic and mechanical properties of the infected erythrocytes during maturation.

Animals↗

Biochemical analysis of DA strain of Theiler's murine encephalomyelitis virus obtained directly from acutely infected mouse brain.

Growth and neurovirulence of a number of neurotropic viruses show pronounced differences after passage in cell culture compared with continued in vivo passage in the central nervous system. The DA strain of Theiler's murine encephalomyelitis virus provides a model for studying these issues since DA virus grown in mouse brain produces acute neuronal disease in weanling mice, but tissue culture-passed DA virus does not. In addition, DA virus grown in mouse brain has a greater 50% mouse lethal dose/50% tissue culture infective dose ratio than tissue culture-passed DA virus. Comparison of these viruses required the analysis of virus purified directly from infected mouse brain, without tissue culture passage. Capsid proteins from DA virus grown in mouse brain were resolved on sodium dodecyl sulfate-polyacrylamide gels and shown to have the same profile as tissue culture-passed DA virus. Viral RNAs were the same size, with no evidence of defective interfering particle production. Two-dimensional gels of in vitro-labeled RNase T1-digested RNA showed that virus variants were more apparent during acute in vivo passage. These genomic differences may be critical in determining the biological behavior of the virus.

Animals↗

Electron microscopy of Listeria monocytogenes-infected mouse spleen.

Armstrong, B. A. (The University of Kansas, Lawrence), and C. P. Sword. Electron microscopy of Listeria monocytogenes-infected mouse spleen. J. Bacteriol. 91:1346-1355. 1966.-Mouse spleen infected with Listeria monocytogenes was observed during the acute phase of infection; 72 hr after infection, organisms were usually found within phagocytic vacuoles in the cytoplasm of macrophages. These vacuoles, which resembled phagosomes, often contained several organisms as well as varying amounts of amorphous electron-dense material, ferritin-like particles, membrane fragments, and vesicles of varying density. Breakdown of vacuolar membranes appeared to be accompanied by damage to the host cell cytoplasm. Nuclear membrane damage was occasionally observed when phagocytic vacuoles were close to the nucleus.

Animals↗

The cytoskeleton of murine leukemia virus (MuLV)-infected mouse fibroblasts as observed under varying conditions of formaldehyde fixation.

Mouse fibroblasts chronically infected with Moloney murine leukemia virus (MuLV) were fixed using variable amounts of formaldehyde, then examined by indirect immunofluorescence light microscopy. Several antisera were employed to detect both external and internal antigens associated with the cells, eg, MuLV gp70, tubulin, vimentin, and actin. Our results indicate that the cell membranes could be partially permeabilized to IgG molecules directed against the three cytoskeletal antigens only after 3.7%, but not 1%, formaldehyde treatment. Complete permeabilization was achieved by subsequent acetone treatment of cells after 3.7% formaldehyde fixation. In such cells, normal-appearing cytoskeletal networks of microtubules and intermediate filaments were observed. Stress fibers were also seen; however, they appeared less numerous and thinner than those of uninfected mouse fibroblasts. Further, a significant amounts of F-actin fluorescence was localized in granules in the cytoplasm of infected cells. Similar observations were made using JLS-V9 mouse cells chronically infected with 334C virus, another MuLV. These results taken together suggest that subtle differences exist in the organization of actin within MuLV-infected and uninfected mouse fibroblasts.

Actins↗

Comparative chemotherapeutic activity of new fluorinated 4-quinolones and standard agents against a variety of bacteria in a mouse infection model.

The new fluorinated 4-quinolones appear to represent orally effective alternatives to parenteral and oral agents currently in use. A number of new fluorinated 4-quinolones were compared in acute systemic mouse-infection models with various Gram-positive cocci (streptococci and staphylococci), Enterobacteriaceae and Pseudomonas aeruginosa. Also included were standard oral and parenteral antimicrobial agents. CI-934 was the most potent quinolone in infections induced by Streptococcus pyogenes and Str. pneumoniae. CI-934, ciprofloxacin, enoxacin, norfloxacin, ofloxacin and pefloxacin were as effective as or superior to standard oral agents currently utilized in infections induced by the Enterobacteriaceae and staphylococci. They were active against antibiotic-susceptible strains and strains resistant to beta-lactams and gentamicin. Most were also quite potent against systemic P. aeruginosa mouse infections. These studies indicate good chemotherapeutic potential for the new generation fluorinated 4-quinolones in infections induced by the staphylococci, streptococci, Enterobacteriaceae and P. aeruginosa, including strains resistant to standard antimicrobial agents.

4-Quinolones↗

Indiscriminate degradation of RNAs in interferon-treated, vaccinia virus-infected mouse L cells.

In this report we used Northern blot hybridization analysis to characterize the fate of several species of viral RNA transcribed from internal and terminal regions of vaccinia DNA in interferon-treated, infected mouse L cells grown in suspension. All species of viral RNAs were expressed but were reduced in amount. Larger-sized RNAs were reduced more than smaller-sized RNAs. This reduction appears to be related to the activation of the interferon-mediated double-stranded RNA-dependent 2-5A synthetase-endoribonuclease system, as the rRNA cleavage pattern characteristic of this system was observed early in infection and in cell extracts in response to exogenous 2-5A. Thus, in interferon-treated, vaccinia-infected mouse L cells in suspension, there is indiscriminate degradation of viral and cellular RNAs, and this RNA breakdown might play a role in the interferon-mediated inhibition of protein synthesis.

Animals↗

Enhanced fusion capacity of malaria (Plasmodium berghei)-infected mouse red cells.

The capacity of normal and malaria-infected mouse red cells to undergo fusion was investigated by phase contrast microscopy. The fusion of mouse red cells induced by 50% w/w poly(ethylene glycol)-6000 in the presence of Ca+2 is enhanced by P. berghei infection. Cells carrying parasites in the ring form stage and early trophozoite stage show slightly higher fusion induced by dimethyl sulphoxide and Ca+2 than those carrying parasites in trophozoite and schizont stages.

Animals↗

Regulation of Ca2+/calmodulin kinase II inhibitor alpha (CaMKIINalpha) in virus-infected mouse brain.

The alpha and beta isoforms of rat Ca(2+)/calmodulin kinase II inhibitor (CaMKIINalpha/beta) expressed in brain or brain and testis, respectively, are potent inhibitors of Ca(2+)/calmodulin kinase II (CaMKII) in vitro. However, the regulation or function of CaMKIINalpha/beta in the central nervous system (CNS) is not known. In this study, we demonstrate that mouse CaMKIINalpha gene encodes a 2.9kb brain-specific transcript whose expression is downregulated in mouse brain during Japanese encephalitis virus (JEV) and rabies virus infection. The downregulation is specific for CaMKIINalpha but not CaMKIINbeta mRNA. In addition to these changes in CaMKIINalpha mRNA, distinct changes are also observed in the phosphorylation as well as subcellular localization of CaMKIIalpha leading to an increase in cytosolic CaMKII activity in JEV-infected mouse brain. The differential regulation of CaMKIIalpha and CaMKIINalpha during JEV infection suggests a possible role for these proteins in viral infection and/or virus-induced neuropathogenesis in the CNS.

Amino Acid Sequence↗

Protective immune response in mice immunized with antigens from Trypanosoma gambiense-infected mouse blood.

Immunogenicity and property of antigens obtained from Trypanosoma gambiense-infected mouse blood (IMP) were examined. A strong vaccine effect against intravenous challenges with 3 x10(3) parasites given on study day 3, 5, or 14 (day 0 = immunization) was observed in mice immunized with a combination of IMP (2 mg protein/mouse) and Freund's complete adjuvant (FCA). But when the challenge was given on day 21 or 30, per cent survival in mice dropped to the 20- and 40-per cent level, respectively. Among fractioned components of IMP, IMP-1, IMP-2, and imp-3, by gel filtration with Sephadex G-200, all of the mice immunized with IMP-1 antigen alone or together with FCA and challenged on day 5 were able to conquer intraperitoneal challenges with 1 x10(2) parasites. Mice immunized with IMP-2 or IMP-3 died within 6 days after challenge. Moreover, protection efficacy shown by IMP-1p (144,000 xg sediment of IMP-1) antigen in mice was similar to that by IMP and IMP-1 antigens. IMP-3 yielded a single precipitin line against mouse anti-IMP serum by Ouchterlony double diffusion method but this response was eliminated when the antiserum was absorbed by IMP-1p. No precipitin line was identified between mouse anti-IMP serum and IMP-1 or IMP-2. From electron microscopic observations, elements of IMP-1 and IMP-1p are possibly corresponded to the fragments of filopodia of the parasites.

Agglutination Tests↗

Ex vivo model of leukocyte migration into herpes simplex virus-infected mouse corneas.

We are investigating neutrophilic infiltration into herpes simplex virus type 1 (HSV-1)-infected mouse corneas. Using a novel ex vivo corneal migration assay, we demonstrated two waves of neutrophil chemotaxis in HSV-1-infected corneas. An early chemotactic gradient developed within 48 h in concert with the appearance of HSV-1 epithelial lesions, peaked by 4 days post-infection (p.i.), and degraded by 6-8 days p.i.. A second chemotactic gradient appeared 10 days after infection, just before the initiation of chronic inflammation. The gradient was maintained in corneas that developed inflammation but rapidly degraded in corneas that failed to develop inflammation. The early chemotactic gradient was established in the absence of T lymphocytes, but the second chemotactic gradient was virtually abrogated by T cell depletion. We conclude that HSV-1 infection induces two temporally separated neutrophil chemotactic gradients in the mouse cornea: an early, transient, T cell-independent gradient; and a later, chronic, T cell-dependent gradient.

Animals↗

Natural immunity in the oncornavirus-infected mouse.

BALB/cfC3H females neonatally infected with mammary tumor virus (MTV) possess spleen cells capable of significant cytotoxic activity against target MTV-induced mammary tumor cells in microcytotoxicity assay. Spleen cells from supposedly MTV-free BALB/c females are also specifically reactive, as a result of horizontal transmission of MTV antigens. No quantitative differences in reactivity exist between females not susceptible to the development of mammary tumors (BALB/c) and those that may develop or already have tumors (multiparous BALB/cfC3H). However, there are qualitative differences in response between those at risk and those not at risk.

Age Factors↗

A single hamster PrP amino acid blocks conversion to protease-resistant PrP in scrapie-infected mouse neuroblastoma cells.

Neurodegeneration caused by the transmissible spongiform encephalopathies is associated with the conversion of a normal host protein, PrP-sen, into an abnormal aggregated protease-resistant form, PrP-res. In scrapie-infected mouse neuroblastoma cells, mouse PrP-sen is converted into PrP-res but recombinant hamster PrP-sen expressed in these cells is not. In the present studies, recombinant hamster/mouse PrP-sen molecules were expressed in these scrapie-infected cells to define specific PrP amino acid residues critical for the conversion to PrP-res. The results showed that homology to the region of mouse PrP-sen from amino acid residues 112 to 138 was required for conversion of recombinant PrP-sen to PrP-res in scrapie-infected mouse cells. Furthermore, a single hamster-specific PrP amino acid at residue 138 could inhibit the conversion of the recombinant PrP-sen into PrP-res. The data are consistent with studies in humans which show that specific amino acid residue changes within PrP can influence disease pathogenesis and transmission of transmissible spongiform encephalopathies across species barriers.

Amino Acid Sequence↗

Polyoma virus early and late mRNAs in productively infected mouse 3T6 cells.

We mapped polyoma virus-specific mRNAs isolated from productively infected mouse 3T6 cells on the viral genome by analyzing nuclease S1-resistant RNA-DNA hybrids. The polyoma early mRNAs, which code for the three T antigens, have several 5' ends near 73 map units (m.u.). During the late phase of infection an additional 5' end is found near 71 m.u. All of the major early mRNAs have common 3' ends at 26.01 m.u. There is a minor species of early mRNA with a 3' end at 99.05 m.u. There are two proximal and two distal splice junctions in the early region which are used to generate three different spliced early mRNAs. There are three late mRNAs encoding the three virion proteins, VP1, VP2, and VP3. The late mRNAs have common 3' ends at 25.34 m.u. The late mRNAs have heterogeneous 5' leader sequences derived from the region between 65.53 and 68.42 m.u. The leader sequences are joined to the bodies of the messages coding for VP2, VP3, and VP1 at 66.59, 59.62, and 48.57 m.u., respectively. These results confirm and extend previous analyses of the fine structure of polyoma mRNAs.

Animals↗

Comparative effects of amoxycillin and ampicillin in the treatment of experimental mouse infections.

Amoxycillin was significantly more active than ampicillin in the treatment of intraperitoneal mouse infections when administered by oral and parenteral routes, although the causal bacteria were equally susceptible in vitro to the two penicillins. Amoxycillin produced higher antibiotic blood concentrations in mice than ampicillin after oral administration, and this was a possible explanation for the superior oral activity of amoxycillin. In contrast, antibiotic blood concentrations were the same for both compounds after subcutaneous injection, but it was demonstrated that amoxycillin was more effective than ampicillin by this route in reducing bacterial counts in the peritoneal cavity and in the blood of mice infected with Escherichia coli. Amoxycillin was also significantly more active than ampicillin in the treatment of infection by intraperitoneal dosing as a result of greater bactericidal activity in infected mice together with the production of higher antibiotic blood levels. The results of these studies on the effects of parental treatment of experimental infections with the two penicillins show that the superior chemotherapeutic activity of amoxycillin was associated with the greater bactericidal activity of amoxycillin in vivo and with differences in the distribution of the two penicillins in the infected animal.

Amoxicillin↗