Calcium environment in bone mineral determined by EXAFS spectroscopy

Summary Extended X-ray absorption fine structure (EXAFS) spectra were recorded, above the Ca K edge, from powdered mouse femurs. Spectra were interpreted on the basis of a model developed previously to explain the features of the EXAFS spectrum of fully crystalline hydroxyapatite. Eight shells of atoms surrounding Ca out of 0.57 nm were required to explain the appearance of the EXAFS spectrum of bone. Shell radii and Debye-Waller factors were systematically varied to obtain the best fit between observed and theoretical spectra, calculated using exact spherical wave theory. The results were closely similar to those obtained previously from the interpretation of EXAFS spectra from poorly crystalline hydroxyapatite prepared by maturation of amorphous calcium phosphate. However, there appears to be slightly more disorder in bone mineral, perhaps as a result of its accommodating carbonate ions     

Genome sequence based, comparative analysis of the fluorescent amplified fragment length polymorphisms (FAFLP) of tubercle bacilli from seals provides molecular evidence for a new species within the Mycobacterium tuberculosis complex.

Tuberculosis in seals is caused by a member of the Mycobacterium tuberculosis complex referred to as the 'seal bacillus'. Fluorescent amplified-fragment length polymorphism (FAFLP) analysis was applied to isolates from four Australian and six Argentinean seals and compared with FAFLP pattern for standard strains belonging to the M. tuberculosis complex. The FAFLP profiles derived from EcoRI/MseI restricted fragments of blind coded DNA samples differentiated the seal bacillus from other members of the M. tuberculosis complex. According to the phylogenetic analysis performed using FAFLP data, seal bacilli appear to have diverged significantly from other members of the M. tuberculosis complex. We describe the suitability of a panel of 19 highly polymorphic markers for rapid identification and comparative genomic analyses of the seal bacillus strains. It is likely that these bacilli got separated from the M. tuberculosis lineage as a result of different insertion deletion events occurring on a genome wide scale. Our analysis reveals that the seal bacillus and M. bovis are genetically related and therefore, might have originated from a common ancestor. Our data additionally support the hypothesis that seal bacillus occupies a unique taxonomic position within the M. tuberculosis complex.     

Analysis of genomic downsizing on the basis of region-of-difference polymorphism profiling of Mycobacterium tuberculosis patient isolates reveals geographic partitioning

Mycobacterium tuberculosis, the etiological agent of tuberculosis, has lost many coding and noncoding regions in its genome during the course of evolution. We performed region-of-difference (RD) analysis using PCR-based genotyping of 131 M. tuberculosis clinical isolates obtained from four different countries, namely, India, Peru, Libya, and Angola. Our studies revealed that RD patterns are often distinct for strains circulating in specific geographical regions and can be used to trace the descent and spread of an isolate from its original reservoir. We describe our findings, which show that no single isolate from the four countries (n = 131) had all the 15 RDs either deleted or retained. Tuberculosis-specific deletion 1 (TbD1) was found to be conserved in 23% of the Indian isolates, indicating their possible ancient origin. RD9 was the most conserved region, RD11 was predominantly deleted, and RD6 was the most variable among the isolates in our collection irrespective of their geographic region. In contrast to earlier reports, our results demonstrate that the deletion of RD1 does not correlate with a decrease in the virulence potential of M. tuberculosis, as Indian isolates (n = 30) examined by us were from diseased individuals and yet had lost the RD1 region. Our results further illustrated that the intactness of the RD5 region may be associated with increased virulence of the organism. This study highlights that the RDs in M. tuberculosis genomes are geographically distributed and specific and may possibly be associated with virulence spectrum.     

Real-time transesophageal echocardiography for intraoperative surveillance of patients with renal cell carcinoma and vena caval extension undergoing radical nephrectomy

PURPOSE: Vena caval tumor thrombus associated with renal cell carcinoma occurs in 4 to 10% of all renal tumors. There is significant operative morbidity and mortality in removing these tumors. We investigate the use of real-time transesophageal echocardiography intraoperatively and to identify tumor thrombus migration and air embolus, which are 2 potentially fatal complications of this procedure. MATERIALS AND METHODS: A total of 13 consecutive patients with renal masses and vena caval extension underwent extirpative surgery monitored with real-time transesophageal echocardiography. RESULTS: In 11 cases the involved kidney and tumor thrombus were removed without morbidity and no evidence of tumor migration or air embolus. Transesophageal echocardiography revealed a 5 cm. tumor thrombus in the right atrium which was removed by immediate atriotomy in 1 of the remaining 2 cases, and a large volume of air in the right atrium that was percutaneously evacuated in the other. These intraoperative complications were unsuspected and only recognized due to the use of transesophageal echocardiography. CONCLUSIONS: Real-time transesophageal echocardiography is a useful adjunct to surgery in patients with renal cell carcinoma and vena caval extension. Transesophageal echocardiography facilitates identification of tumor thrombus migration and air embolization, which are potentially fatal complications, and allows for immediate intraoperative intervention.     

Absence of ATM truncations in patients with severe acute radiation reactions

Purpose: Severe acute toxicity limits the effective use of radiotherapy in patients who are radiosensitive, and it is not usually possible to identify these radiohypersensitive (R-H) individuals before treatment commences. Five such R-H patients were detected over a 3-year period. We undertook this study to determine whether the severe acute radiohypersensitivity of these five individuals showed any correlation with cellular and molecular parameters known to be abnormal in radiosensitivity-related syndromes such as ataxia–telangiectasia (A-T).

Methods and Materials: Lymphoblastoid cells were isolated from fresh blood from the 5 R-H individuals who had previously demonstrated clinical R-H at least 9 months prior to sampling. Lymphoblastoid cell lines (LCLs) were established to determine the extent of postradiation chromosomal aberrations, cell cycle delay, cell proliferation, and tumor suppressor p53 protein stabilization. The polymerase chain reaction (PCR) and protein truncation (PTT) assays were used to test for the possibility of mutations in the gene mutated in A-T, termed ATM.

Results: LCLs derived from R-H subjects retained a significantly higher degree of radiation-induced chromosomal aberrations when compared to normal control LCLs. p53 stabilization by ionizing radiation appeared normal in all but one R-H subject. There was no evidence of A-T gene truncation mutations in any of the R-H subjects tested.

Conclusions: All R-H subjects in this study had their cellular radiosensitivity confirmed by the chromosomal aberration assay. Delayed p53 stabilization at 4 hours postirradiation in one R-H subject suggested that different etiologies may apply in the radiohypersensitivity investigated in this study.     

An Evaluation of the Performance of Direct Agglutination Test on Filter Paper Blood Sample for the Diagnosis of Visceral Leishmaniasis

The attack phase of the visceral leishmaniasis (VL) elimination program in Bangladesh aims to decrease the burden of VL incidence from close to 20 cases to less than one case per 10,000 at sub-district level. The consolidation phase will aim to confirm no increase in VL in endemic areas through active surveillance. During this phase, a reliable diagnostic tool for mass screening is required. Here, we report the diagnostic sensitivity and specificity of a filter paper-based agglutination test (FP-DAT) for diagnosis of VL in patients admitted to an upazila health complex in Mymensingh, a VL-endemic region of Bangladesh. The sensitivity of both the conventional direct agglutination test (DAT) and FP-DAT were 100% and 96%, respectively. The specificity of both assays was 100%. However, when the performances of the two assays were compared using McNamar''s test, neither the sensitivity nor the specificity of the FP-DAT differed significantly from conventional DAT.