Cloning, Expression, and Sequencing of a Cell Surface Antigen Containing a Leucine-Rich Repeat Motif from Bacteroides forsythus ATCC 43037

Bacteroides forsythus is a recently recognized human periodontopathogen associated with advanced, as well as recurrent, periodontitis. However, very little is known about the mechanism of pathogenesis of this organism. The present study was undertaken to identify the surface molecules of this bacterium that may play roles in its adherence to oral tissues or triggering of a host immune response(s). The gene (bspA) encoding a cell surface-associated protein of B. forsythus with an apparent molecular mass of 98 kDa was isolated by immunoscreening of a B. forsythus gene library constructed in a lambda ZAP II vector. The encoded 98-kDa protein (BspA) contains 14 complete repeats of 23 amino acid residues that show partial homology to leucine-rich repeat motifs. A recombinant protein containing the repeat region was expressed in Escherichia coli, purified, and utilized for antibody production, as well as in vitro binding studies. The purified recombinant protein bound strongly to fibronectin and fibrinogen in a dose-dependent manner and further inhibited the binding of B. forsythus cells to these extracellular matrix (ECM) components. In addition, adult patients with B. forsythus-associated periodontitis expressed specific antibodies against the BspA protein. We report here the cloning and expression of an immunogenic cell surface-associated protein (BspA) of B. forsythus and speculate that it mediates the binding of bacteria to ECM components and clotting factors (fibronectin and fibrinogen, respectively), which may be important in the colonization of the oral cavity by this bacterium and is also a target for the host immune response.     

Intracranial olfactory neuroblastoma: evidence for olfactory epithelial origin.

AIMS: To determine the possible histogenesis of the intracranial variant of olfactory neuroblastoma. METHODS: Four specimens from three cases of intracranial olfactory neuroblastoma were studied by light microscopy and immuno-histochemistry, and electron microscopy in two cases. RESULTS: Light microscopical examination showed small cell tumour with additional features of epithelioid cells in one case and ganglion cells in another. Olfactory and Homer-Wright rosettes were present. All the specimens showed a uniform positive reaction to neurone specific enolase, S-100, and cytokeratin antibodies. Glial fibrillary acidic protein was absent. The salient electron microscopic features were the presence of cell junctions, cytoplasmic intermediate filaments, basal bodies and cytolasmic processes. Dense cored vesicles were absent. CONCLUSIONS: The results strongly support the view that intracranial olfactory neuroblastomas are of olfactory epithelial origin and differ from conventional neuroblastomas.     

The GAA triplet-repeat sequence in Friedreich ataxia shows a high level of somatic instability in vivo,with a significant predilection for large contractions

Friedreich ataxia is commonly caused by large expansions of a GAA triplet-repeat (GAA-TR) sequence in the first intron of the FRDA gene. We used small-pool PCR to analyze somatic variability among 7190 individual FRDA molecules from peripheral blood DNA of subjects carrying 12 different expanded alleles, ranging in size from 241 to 1105 triplets. Expanded alleles showed a length-dependent increase in somatic variability, with mutation loads ranging from 47% to 78%. We noted a strong contraction bias among long alleles (>500 triplets), which showed a 4-fold higher frequency of large contractions versus expansions. Some contractions were very large; of all somatic mutations scored, approximately 5% involved contractions of >50% of the original allele length, and 0.29% involved complete reversion to the normal/premutation length (< or =60 triplets). These observations contrast sharply with the strong expansion bias seen in expanded CTG triplet repeats in myotonic dystrophy. No somatic variability was detected in >6000 individual FRDA molecules analyzed from 15 normal alleles (8-25 triplets). A premutation allele with 44 uninterrupted GAA repeats was found to be unstable, ranging in size from 6 to 113 triplets, thus establishing the threshold for somatic instability between 26 and 44 GAA triplets. Analysis of an additional 7850 FRDA molecules from serially passaged lymphoblastoid cell lines carrying nine expanded alleles (132-933 triplets) showed very low mutation loads, ranging from 0% to 6.2%. Our data indicate that expanded GAA-TR alleles in Friedreich ataxia are highly mutable and have a natural tendency to contract in vivo, and that these properties depend on multiple factors, including DNA sequence, triplet-repeat length and unknown cell-type-specific factors.     

Low delay rate adaptive pacemaker using FPGA embedded piezoelectric sensor

Abstract

This paper presents the hardware implementation of low delay, power-efficient, rate-adaptive dual-chamber pacemaker (RDPM) using a piezoelectric sensor. Rate adaptive pacemaker has the ability to sense the patient’s activity by means of some special sensors and it controls the pacing rate according to the patient’s activity. Ideally, there should be no delay between sensing and the subsequent pacing operation performed by the pacemaker. However, delay in the responses of various components in the circuitry produces an accumulative delay effect in any practical circuit. Physical activity and the physiological needs of the patient can be easily adapted by the rate-responsive pacemakers using a wide range of sensor information. The piezo-electric sensor recognises the pressure on human muscles because of physical activity and converts it to an electrical signal, which is received by the pulse generator of the pacemaker. When the patient is in the rest mode, the heart rate is the only parameter that is to be detected by the pacemaker. Thus, the heart rate and the physical activity both are the inevitable parameters for the design of RDPM. Performance analysis of the proposed RDPM shows a significant reduction in the delay between sensing and pacing. Device utility analysis shows that the proposed design not only requires lesser memory but also reduces the number of components on the chip. Therefore, it becomes very clear that the proposed pacemaker design will consume much lesser power.     

Evoked potential changes in ischaemic myelopathy

Somatosensory evoked potentials are employed in the intraoperative monitoring but there is paucity of information about the motor evoked potentials (MEP) in spinal cord ischaemia. Two patients aged 9 and 4 years developed paraplegia following surgery for coarctation of aorta. The aortic cross clamping time in these patients was 100 min and 30 min. respectively. The patient with longer clamping time had flaccid paraplegia and lower limb MEPs were not recordable; whereas the patient with shorter clamping time had spastic paraparesis and prolonged CMCT to right lower limb. The latter patient improved significantly. Tibial SEPs were normal in both the patients. Our results suggest that MEP may have a greater role than SEP in documenting and prognosticating ischaemic myelopathy.