Allele-Specific PCR Method Based on pncA and oxyR Sequences for Distinguishing Mycobacterium bovis from Mycobacterium tuberculosis: Intraspecific M. bovis pncA Sequence Polymorphism

An allele-specific amplification method based on two genetic polymorphisms to differentiate Mycobacterium tuberculosis from Mycobacterium bovis was tested. Based on the differences found at position 169 in the pncA genes from M. tuberculosis and M. bovis, a PCR system which was able to differentiate most of the 237 M. tuberculosis complex isolates tested in one of the two species was developed. All 121 M. tuberculosis strains showed the expected base (cytosine) at position 169. Most of the M. bovis isolates had a guanine at the cited position. Nevertheless, 18 of the 116 M. bovis isolates, all of them goat isolates, showed the pncA polymorphism specific to M. tuberculosis. These results suggest that goat M. bovis may be the nicotinamidase-missing link at the origin of the M. tuberculosis species. Based on the polymorphism found at position 285 in the oxyR gene, the same system was used to differentiate M. tuberculosis from M. bovis. In this case, DNAs from all 121 M. tuberculosis isolates had the expected base (guanine) at this position. In addition, all 116 M. bovis isolates, including those from goats, showed the identical polymorphism (adenine). The oxyR allele-specific amplification method can differentiate M. bovis from M. tuberculosis, is rapid (results can be obtained in less than 3 h), and is easy to perform.     

A fucose-containing epitope potentially involved in gamete interaction on the human zona pellucida

The oligosaccharide moiety of human, porcine and bovine zonaepellucidae was studied with lectins and monoclonal antibodiesspecific for tri- or tetra-saccharidic epitopes containing atleast one terminal -L-fucose. Animal eggs were collected fromfollicular aspirates, human eggs were collected from in-vitrofertilization and embryo transfer programmes and pooled intosix groups. By direct immunofluorescence, the lectins reactivitywas detected for the animal or the human zonae pools in thesame way. Reactivity of Aleuria aurantia lectin demonstratedthe presence of –L-fucose terminal residues in the zonaefrom the three species studied. By indirect immunofluorescence,the 2–25 antibody reactivity was detected in every poolof human zonae whereas there was no evidence of any antibodyreactivity on animal zonae. Using an anti-Lewis-b blood groupantibody (2–25), we observed expression of this antigenas an intrinsic component of the human zona pellucida, independentlyof patients'Lewis red blood cell phenotypes. Antibody 2–25inhibited the sperm–atozoa-zona binding in a hemizonaassay, suggesting that this fucose-containing antigen couldbe part of a sperm-zona receptor.     

Correlation of proliferation of lung epithelium with intramuscular sensitization and complement-fixing antibody to respiratory syncytial virus in the golden hamster.

Intramuscular sensitization of hamsters with several forms of respiratory syncytial virus (RSv) caused proliferation of lung epithelium. In contrast, intranasal injection of live virus rarely resulted in this phenomenon. A correlation existed between proliferation of lung epithelium and presence of complement-fixing antibody, but not between lung disease and delayed skin reactions. Complement-fixing antibody to RSv was found to be independent of the influence of the thymus.     

Contrasting roles of DAP10 and KARAP/DAP12 signaling adaptors in activation of the RBL-2H3 leukemic mast cell line

A common feature of hematopoietic activating immunoreceptors resides in their association at the cell surface with transmembrane signaling adaptors. Several adaptors, such as the CD3 molecules, FcRgamma and KARAP/DAP12, harbor intracytoplasmic immunoreceptor tyrosine-based activation motifs (ITAM) that activate Syk-family protein tyrosine kinases. In contrast, another transmembrane adaptor, DAP10, bears a YxxM motif that delivers signals by activation of lipid kinase pathways. We show here that the human signal-regulatory protein SIRPbeta1 can associate with both DAP10 and KARAP/DAP12 in a model of RBL-2H3 cell transfectants. In association with KARAP/DAP12, SIRPbeta1 complexes are capable of inducing serotonin release and tumor necrosis factor (TNF) secretion. By contrast,in the absence of KARAP/DAP12, engagement of SIRPbeta1:DAP10 complexes does not lead to detectable serotonin release or TNF secretion by RBL-2H3 transfectants. However, triggering of SIRPbeta1:DAP10 complexes co-stimulates RBL-2H3 effector function induced by sub-optimal stimulation of the endogenous FcepsilonRI complex. Therefore, we report here a cellular model in which the association of a cell surface receptor with various signaling adaptors dictates the co-stimulatory or the direct stimulatory properties of the complex.     

A second case of neonatal alloimmune thrombocytopenia associated with anti-PlA2 (Zwb) antibodies

A case of neonatal alloimmune thrombocytopenia due to fetomaternal incompatibility against the platelet specific antigen PlA2 (Zwb) is described. Anti-PlA2 antibodies were present in the maternal serum together with anti-HLA antibodies. Initial identification of the platelet-specific antibodies was achieved by using chloroquine-treated PlA1(+) and PlA1(-) platelets. Antibody specificity was later confirmed using a panel of platelets of known phenotype.     

A self-setting single-component calcium phosphate cement

Following an original synthesis route, we have prepared a single-component calcium phosphate apatite-like powder which settles and hardens when mixed with deionized water in an approximative 1.2 g:1 ml ratio. This paper describes the first physico-chemical studies and characterizations of the material. Observations of its in vitro behavior show a slight volume contraction and toxicity against fibroblasts bone marrow cells on disks of compacted powder. It is suggested that after an improvement of the powder characters such as grain size, and the choice of another hardening liquid, to name a few, this material should be a potential--or an ingredient of--bone cement.     

In vivo and in vitro models of demyelinating diseases. II. Persistence and host-regulated thermosensitivity in cells of neural derivation infected with mouse hepatitis and measles viruses.

Following inoculation of continuous cell lines of neural and other derivations, persistent infections are established with facility by mouse hepatitis and measles viruses. This occurs equally with the prototype MHV₃ and its neurotropic variant JHM as well as with the Edmonston vaccine and SSPE Hallé measles variants. In almost every instance that the infection becomes persistent at 32.5°, virus replication is found to be thermosensitive at 39.5°; however, progeny virus derived from such infections at 32.5° is itself thermostable when replicating in the indicator, fully permissive cell lines. The new data, therefore, reveal the existence of a host-conferred interrelationship between persistence and virus restriction at elevated temperature. They indicate that the two agents with neurotropic potential, when they become established as pathogens in the nervous system, could be under close host cell regulation involving as yet unknown mechanisms.     

Unusual giant cell tumor arising in a male breast

An unusual giant cell tumor of the breast of a 72 year old man is reported. The microscopic and ultrastructural features of the tumor are presented in detail. Unusual and previously unreported myofibroblastic and myoepithelial differentiation of the spindle cell component is described. The possible histogenesis of the tumor is discussed.