Ewingella americana: recurrent pseudobacteremia from a persistent environmental reservoir.

From September 1981 through April 1984, 20 patients at one hospital were identified with Ewingella americana pseudobacteremia. Case-control studies demonstrated an association between having a positive blood culture for E. americana and having blood for culture obtained simultaneously with blood obtained for coagulation studies (15 of 19 case patients versus 4 of 38 controls; P = 4.5 X 10(-7)). Review of blood-drawing procedures showed that blood for coagulation studies and culture was drawn with the same syringe, and coagulation tubes were filled before blood culture tubes. Some phlebotomists were not using new sterile needles to inoculate blood culture bottles. Collection tubes for coagulation studies were prepared in the hospital, and E. americana was isolated from all 52 unused coagulation tubes tested. Solutions prepared in the hospital may constitute a persistent inanimate environmental reservoir for this uncommon microorganism. Pseudobacteremia can result in unnecessary antimicrobial therapy for some patients, incurring the risks of adverse drug reactions, selection of drug-resistant bacteria, and increased health care costs.     

Polyomavirus Models of Brain Infection and the Pathogenesis of Multiple Sclerosis

Multiple sclerosis (MS) is generally considered to be an autoimmune disorder with myelin as the target and with several unidentified viruses playing ancillary roles, possibly through molecular mimicry. Although this paradigm has led to important progress on potential mechanisms of myelin loss, neither a target antigen in myelin nor a triggering mechanism has yet been identified, leaving the etiology of MS still unknown. Animal models of viral demyelination and studies showing that JC virus (JCV), the polyomavirus which causes progressive multifocal leukoencephalopathy (PML), may be latent in some normal human brains suggest another possibility. A host immune response targeting proteins expressed at low levels from viral DNA latent in the central nervous system (CNS) might underlie a focal demyelinating disease such as MS. A shift from autoimmunity to a latent-virus model is not a trivial substitution of target antigens. This shift would expand the search for a definitive laboratory test for MS and could lead to improved therapeutic and preventive approaches.     

Intermittent preventive sulfadoxine-pyrimethamine treatment of primigravidae reduces levels of plasma immunoglobulin G, which protects against pregnancy-associated Plasmodium falciparum malaria

Pregnancy-associated malaria (PAM) is an important cause of maternal and neonatal suffering. It is caused by Plasmodium falciparum capable of inhabiting the placenta through expression of particular variant surface antigens (VSA) with affinity for proteoglycans such as chondroitin sulfate A. Protective immunity to PAM develops following exposure to parasites inhabiting the placenta, and primigravidae are therefore particularly susceptible to PAM. The adverse consequences of PAM in primigravidae are preventable by intermittent preventive treatment (IPTp), where women are given antimalarials at specified intervals during pregnancy, but this may interfere with acquisition of protective PAM immunity. We found that Kenyan primigravidae receiving sulfadoxine-pyrimethamine IPTp had significantly lower levels of immunoglobulin G (IgG) with specificity for the type of parasite-encoded VSA-called VSA(PAM)-that specifically mediate protection against PAM than did women receiving a placebo. VSA(PAM)-specific IgG levels depended on the number of IPTp doses received and were sufficiently low to be of clinical concern among multidose recipients. Our data suggest that IPTp should be extended to women of all parities, in line with current World Health Organization recommendations.     

Differential activation of human and guinea pig complement by pentameric and hexameric IgM

Human and mouse IgM can be polymerized as a hexamer in addition to a pentamer. Our previous work with mouse IgM measured activation of guinea pig complement by highly enriched preparations of hexamer and pentamer and showed that hexamer is >100-fold more active than pentamer. In this report pentamer and hexamer were compared for their capacity to activate complement in a homogeneic system, i.e. chimeric mouse V/human Cmu IgM pentamer and hexamer were assayed separately for their capacity to activate human (and guinea pig) complement. In both the homogeneic and the xenogeneic systems hexamer was more active than pentamer, but the magnitude of the difference between hexamer and pentamer depended on the complement source. Whereas chimeric hexamer activated guinea pig complement >100-fold more efficiently than did chimeric pentamer, this hexamer was only 4-13-fold more active than pentamer when assayed with human complement. Similarly, mouse hexamer, which was >100-fold more active than mouse pentamer with guinea pig complement, was only approximately 2-fold more active than mouse pentamer with human complement. Mouse hexameric and pentameric IgM were each approximately 20-fold more active with human complement than were the corresponding chimeric isoforms of IgM.     

Summary of complementation groups of UV-sensitive CHO cell mutants isolated by large-scale screening

A summary is given for the lineage and complementation groupassignments of 153 UV-sensitive mutants of the CHO AA8 cellline. The distribution of mutants among six complementationgroups was highly non-random, with the great majority of theisolates belonging to groups 1 and 2. This asymmetry is consistentwith the known hemizygosity of these two linked loci in CHOcells. The relative numbers of mutants induced in group 2 wasfound to depend greatly on the type of mutagen used. Mutagenesiswith UV radiation, ethyl methanesulfonate (EMS), N-methyl-N'-nitro-N-nitroso-guanidine and 7-bromomethylbenz[a]anthraceneproduced high frequencies of group 2 mutants. In contrast, ICR170and ICR191, which are thought to produce mostly frameshift mutations,yielded very few mutants in group 2. These results are of particularimportance in light of the recent finding that the human ERCC2gene, which corrects group 2 mutants, has very strong homologywith the yeast gene RAD3. RAD3 is an essential gene for viabilityin yeast, and the low recovery of group 2 mutants using theframeshift agents strongly suggests that frameshift mutationstend to be lethal in the hamster ERCC2 locus. Several mutagen-sensitivedouble mutants were isolated in two-step selections from EMS-,mitomycin C- or UV-sensitive parental cells, including the lineUVU1, the first mammalian line with two mutations that affectUV sensitivity. The first mutation inactivated excision repair,and the second mutation appears to have affected some otherrecovery process. UVU1 should be useful for studying recoveryprocesses that are separate from nucleotide excision repair. ¹To whom correspondence should be addressed     

Kidney antigens active in isoimmunization: I. The antigenic diversity

Isoimmunization of rabbits with kidney extracts and Freund's adjuvant leads to the development of responses to a multiplicity of autoantigens. If a large number of kidneys is pooled, there is increased probability that responses directed toward isoantigens can also be detected. Such immunization thus results largely in antisera that possess multiple specificities, but the amount of antibody directed toward each specific antigen varies from one antiserum to another.

Advantage can be taken of these uneven responses by different animals, to enumerate the variety of responses to uniquely separable sets of determinants, by carefully selecting the antisera. In immunodiffusion studies, lines for such separable antigens can be sorted in an unambiguous manner. A total of five such antigens has now been found in kidney, three of which are kidney-specific and two of which are shared with liver. A sixth antigen, separable from liver, may be only a minor constituent of kidney, and was not directly detected in kidney extracts. It is important that fresh concentrated extracts of kidney homogenates be used. The detection of isoantigens requires concentrated extracts prepared from individual kidneys.

     

Nucleation of calcium phosphate by surface-bound extracellular matrix

The native extracellular matrix (ECM) laid down on silicon and titanium surfaces by osteoblast-like SAOS-2 cells was exposed by selective removal of cells. This type of material surface ECM-Si, ECM-Ti was shown to promote the nucleation of calcium phosphate from a simulated body fluid (SBF). Microscopic and spectroscopic results revealed the effect was associated with a collagen fiber-free extracellular matrix.