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Biomedical subjects

M Bansal

Publications and source records attributed to M Bansal.

At least 19 recordsLinked to original sources

Hairpin and parallel quartet structures for telomeric sequences.

The role of thymine residues in the formation of G-quartet structures for telomeric sequences has been investigated using model oligonucleotides of the type d(G4TnG4), with n = 1-4. Sequences d(G4T3G4) and d(G4T4G4) adopt a G-quartet structure formed by hairpin dimerization in 70 mM NaCl as judged by a characteristic circular dichroism signature with a 295 nm positive and 265 nm negative bands while d(G4TG4) adopts a parallel G-quartet structure like d(G12) which exhibits a strong positive band at 260 nm and a negative band at 240 nm. The sequence d(G4T2G4) exhibits a mixture of both conformations. The stability of hairpin G-quartet structures decreases with decrease in the number of intervening thymine residues. Potassium permanganate, a single strand specific probe has been used to establish the presence of loops composed of T residues in the hairpin G quartet structures formed by the oligonucleotides d(G4TnG4) with n = 2-4 in 70 mM NaCl. The formation of hairpin G quartet structure for the above sequences is further supported by the enhanced electrophoretic mobility observed on non-denaturing polyacrylamide gels. Human telomeric sequence d(TTAGGG)4 which showed enhanced electrophoretic mobility like Tetrahymena telomeric sequence d(T2G4)4 also exhibited a characteristic CD spectrum for a folded-back G-quartet structure. A detailed model for G-quartet structure involving hairpin dimer with alternating syn-anti-syn-anti conformation for the guanine residues both along the chain as well as around the G tetrad with at least two thymine residues in the loop is proposed. Intermolecular association of short telomeric sequences reported here provides a possible model for chromosomal pairing.

Animals

Double helix conformation, groove dimensions and ligand binding potential of a G/C stretch in B-DNA.

The self-complementary DNA fragment CCGGCGCCGG crystallizes in the rhombohedral space group R3 with unit cell parameters a = 54.07 A and c = 44.59 A. The structure has been determined by X-ray diffraction methods at 2.2 A resolution and refined to an R value of 16.7%. In the crystal, the decamer forms B-DNA double helices with characteristic groove dimensions: compared with B-DNA of random sequence, the minor groove is wide and deep and the major groove is rather shallow. Local base pair geometries and stacking patterns are within the range commonly observed in B-DNA crystal structures. The duplex bears no resemblance to A-form DNA as might have been expected for a sequence with only GC base pairs. The shallow major groove permits an unusual crystal packing pattern with several direct intermolecular hydrogen bonds between phosphate oxygens and cytosine amino groups. In addition, decameric duplexes form quasi-infinite double helices in the crystal by end-to-end stacking. The groove geometries and accessibilities of this molecule as observed in the crystal may be important for the mode of binding of both proteins and drug molecules to G/C stretches in DNA.

Base Sequence

Effect of short-term hypomagnesemia on the chemical and mechanical properties of rat bone.

Magnesium is known to have an essential role in determining the properties of bone, but the way in which Mg exerts its actions remains unclear. Although long-term Mg deficiency is known to produce osteopenia, the effects of short-term Mg deficiency have not been established. To test the hypothesis that Mg deficiency results in an altered pattern of initial mineralization and concomitant altered bone properties, the radiographic, histologic, chemical, and mechanical properties of the bones of rats given a Mg-deficient diet were compared to those of rats pair-fed the same diet supplemented with Mg. Short-term Mg-deficiency in the diet of growing rats produced a significant decrease in both the trabecular bone volume and the mineral content of the newly formed metaphysis, a significant increase in the Ca:P ratio, and a slight, but significant increase in hydroxyapatite crystallite size and/or perfection in the metaphysis. Comparable, but not significant, trends were found in the diaphyses. Metaphyseal bone osteocalcin levels were reduced in the Mg-deficient rats and lipid was more easily extracted from their bones. No detectable alterations in radiographic microstructure were noted. Mechanically, a significant decrease in the maximum three-point bend strength of the femurs of Mg-deficient rats was observed. These data support the hypothesis that short-term Mg deficiency affects the pattern of bone mineral formation.

Animals

Case report 706: Silicone-induced reactive synovitis.

In a patient with a disrupted silicone prosthesis, the presence of a swollen articulation should suggest the presence of silicone-induced reactive synovitis. Diagnosis of the entity rests on the identification of particulate matter by polarized microscopy and/or scanning electron microscopy--studies that will not be performed routinely unless the diagnosis is suggested preoperatively. The case presented is that of a 68-year-old woman in whom a silicone elastomer trapeziometacarpal prosthesis had been implanted 5 years previously. Painful swelling had ensued, and biopsy confirmed the diagnosis.

Aged

Titanium wear debris in failed cemented total hip arthroplasty. An analysis of 71 cases.

Seventy-one cemented total hip arthroplasties (THAs) were reviewed following removal of the all-titanium alloy femoral stem. Fifty-one hips were primary arthroplasties that failed due to aseptic loosening, 8 were previous revisions with aseptic loosening, and 12 were removed for infection. The average duration of service for the three groups was 4.5 years, 5.0 years, and 3.7 years, respectively. Femoral bone loss in aseptically loose, primary THA was graded as severe in 51%, moderate in 24%, and mild in 20%. Femoral endosteolysis was present in 94%, while acetabular osteolysis was seen in 6%. Histological evaluation of tissues from failed primary arthroplasties revealed polymethyl methacrylate debris in 75% of cases, polyethylene debris in 80%, metal debris in 75%, and chronic inflammatory cells in all cases. Metallic debris was not seen in the failed revision cases and in only 17% of the infected cases. Examination of retrieved femoral components revealed burnishing of the head in all cases, while 71% of stems with aseptic loosening were abraded from the cement. Metal levels from 12 cases averaged 2,111 mg/g of dry tissue (range, 60-11,823); synovial fluid levels from 8 other cases averaged 106 mg/l (range, 22-340). While it is not certain whether metallic particles are a primary cause of loosening or are generated secondarily, their presence seems to accelerate bone loss and loosening.

Adult

Groove width and depth of B-DNA structures depend on local variation in slide.

The groove widths of DNA helix, especially minor groove width, are generally believed to be important for recognition of DNA by various types of ligands. It has been postulated earlier that large negative propeller twist, in the AT rich regions compresses the minor groove of duplex DNA. A systematic study has now been carried out by generating models with different values of local doublet and intra-basepair parameters and calculating their minor groove widths. It is found that several local doublet parameters affect the minor groove width but it depends most strongly on the local step parameters roll and slide when each parameter is considered individually. However, a detailed analysis of the various local parameters within the B-DNA family of crystal structures indicates that propeller twist and slide are most strongly correlated with the observed values of minor groove width. The groove depth is also strongly correlated with slide. Thus the local base sequence dependent variations in slide can modify both the groove width and depth and consequently determine the ligand binding properties of DNA.

Base Sequence

Assessment of renal osteodystrophy in hemodialysis patients.

We performed a prospective study of 30 patients undergoing chronic hemodialysis to determine which of 6 generally available diagnostic procedures provided the most useful information for the assessment of bone disease in hemodialysis patients. The 6 procedures were: routine biochemical measurements, N-terminal parathyroid hormone (N-PTH), radiographic analysis of hands and clavicles, bone density determination by dual photon absorptiometry (DPA), deferoxamine stimulation test, and iliac crest bone biopsy. Serum N-PTH was elevated in 83% of patients but was not significantly associated with abnormalities of other biochemical parameters. No significant relationship was demonstrated between biochemical data and radiographic findings or between biochemical data and bone density by DPA. All patients with abnormal DPA had an elevation of N-PTH; therefore, DPA did not reveal any unsuspected disease. Bone biopsies were done in 20 patients and findings in each were consistent with uremic osteodystrophy, including osteitis fibrosa cystica in 11 patients and aluminum-associated bone disease in 2 patients. Six patients had mixed disease, and 1 patient had osteoporosis. Despite 11 positive deferoxamine tests, bone biopsy revealed aluminum deposition in only 7 of these patients, suggesting extraosseous aluminum accumulation in the remaining 4. Evaluation of the positive and negative predictive accuracies of DPA, x-ray analysis, N-PTH levels, and aluminum bone deposition revealed that normal DPA or x-ray findings do not exclude bone disease, that N-PTH level is a good marker for secondary hyperparathyroidism, and that a negative deferoxamine test excludes aluminum-associated bone disease. Discriminant analysis also reinforced these conclusions.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorptiometry, Photon

Metallic debris in femoral endosteolysis in failed cemented total hip arthroplasties.

A prospective study was undertaken to quantitate metallic and cement debris in 12 consecutive patients with femoral endosteolysis (FE) and aseptic loosening of a cemented total hip arthroplasty. The mean interval between primary and revision surgery was 9.6 years. The average time to onset of FE was 8.9 years. There were four stems each of cobalt-chromium (Co-Cr), stainless steel (SS), and titanium alloy. At revision, tissue was retrieved from FE, the femoral bone-cement pseudomembrane, and the joint pseudocapsule. Histology of these tissues was studied using light and polarized microscopy. Metal and barium levels were measured by atomic absorption spectrophotometry. A histiocytic reaction and particulate cement debris were seen in every case. Polyethylene wear debris was noted in 11 of 12 cases (92%), and metallic debris in four cases (33%). Detectable metal levels were found in the FE in all cases. Metal levels were on average 2.5 times higher in FE than in femoral pseudomembrane, and 4.2 times higher than in joint pseudocapsule. This difference was statistically significant for the Co-Cr and SS groups. Barium levels in areas of FE were on average 1.7 times and 42.4 times higher than in femoral pseudomembrane and joint pseudocapsule, respectively. The difference seen between the FE and the joint pseudocapsule tissue was significant for all three alloy groups. The authors' data demonstrated higher metal and barium levels in FE than in the other tissue sites. Polyethylene and cement debris were noted in nearly every case. Cement, polyethylene, and metallic particulate wear debris may contribute to the pathogenesis and progression of FE.

Adult

Cobalt-alloy metal debris in periarticular tissues from total hip revision arthroplasties. Metal contents and associated histologic findings.

To investigate the hypothesis that an association exists between the presence of metallic particulate or ionic debris released from the components of a total hip replacement and the histologic reaction in the surrounding tissues, cobalt, chromium, nickel, and molybdenum levels were measured in periarticular tissue from 22 individuals who had revision surgery. Total tissue content of the four elements (averaged per case) ranged from 2.7 to 250 micrograms of metal per gr of dried tissue (mean, 39 micrograms/gr); however, within each case, the tissue-metal content varied more widely. The highest total tissue-metal contents occurred in cases revised for infection. Fibrosis, histiocytic reaction, hemorrhage, and necrosis were the most frequent histologic findings. Polyethylene and cement particles each appeared in approximately half of all sections, while microscopic metal particles were seen only in tissues from the infected hips. Tissue-metal content did not correlate with the histologic findings, with any of the demographic variables, nor with duration of implantation. Ratios of the individual constituent elements generally reflected the cobalt-chromium alloy composition, suggesting that metal debris was present predominantly as wear particles. Only for tissues with very low metal contents did departures of these ratios indicate the presence of ionic corrosion products. The extremely low metal contents measured in many cement-bone interfacial tissues and the few metal particles found histologically suggest that metal particles may have been less important in inflammatory reaction and loosening than cement or polyethylene particles.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Magnetic resonance imaging in diagnosis of transient osteoporosis of the hip.

The results of magnetic resonance (MR) imaging in six patients with transient osteoporosis of the hip were reviewed. Short TR/TE (repetition time/echo time) images demonstrated diffusely decreased signal intensity in the femoral head and intracapsular region of the femoral neck. Increased signal intensity was noted with progressive T2 weighting. Bone biopsies were performed in four patients. Histologic findings were nonspecific and included fat necrosis, marrow edema, increased bone resorption, and reactive bone formation. Repeat MR scans in two patients, performed six and eight months after the initial scans, showed an almost complete return to normal marrow signal. All patients became asymptomatic without bony deformity. In the appropriate clinical setting, MR scanning can aid in the diagnosis of transient osteoporosis as the cause of a painful hip.

Acute Disease

Antibacterial effects of fibrin glue-antibiotic mixtures.

In the present in vitro study, we investigated the duration of action and antibacterial effects of nonautologous fibrin glue (FG) combined with antibiotics (Abs) including Ciprofloxacin, Teicoplanin, Cefoxitin, and Gentamicin; the effect of FG alone on bacterial growth was also evaluated. The rate of Ab diffusion from combined FG-Ab clots was evaluated by separate elution with pooled human serum (HS) and normal saline (NS); supernatants were removed daily and assayed for active concentrations of each Ab. The effects of FG and combined FG-Ab clots on bacterial growth were evaluated by inoculating brain-heart infusion (BHI) with Staphylococcus aureus, followed by the addition of FG or FG-Ab clots; a separate set of studies was also performed with the addition of mouse lung homogenate (MLH) as well. The addition of Ab to FG clots resulted in continuous diffusion of the Ab into the surrounding HS or NS for up to 5 to 7 days; however, more than two-thirds of the Ab diffused out within 2 to 3 days regardless of the Ab used. The antibacterial effects of FG-Ciprofloxacin and FG-Teicoplanin clots were significant compared to those of FG clots not containing Ab. In addition, the presence of FG clots (in BHI) resulted in a reduction in bacterial growth compared to that of BHI alone. The addition of MLH to BHI resulted in increased bacterial growth, but this effect was inhibited by the presence of combined FG-Ab clots.(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Bacterial Agents

DNA polymorphism and local variation in base-pair orientation: a theoretical rationale.

Basepair stacking calculations have been carried out to understand the conformational polymorphism of DNA and its sequence dependence. The recently developed self-consistent parameter set, which is specially suitable for describing irregular DNA structures, has been used to describe the geometry of a basepair doublet. While for basepairs without any propeller, the favourable stacking patterns do not appear to have very strong features, much more noticeable sequence dependent stacking patterns emerge once a propeller is applied to the basepairs. The absolute minima for most sequences occurs for a doublet geometry close to the B-DNA fibre models. Hence in the B-DNA region, no strong sequence dependent features are found, but the range of doublet geometries observed in the crystal structures generally lie within the low energy contours, obtained from stacking energy calculations. The doublet geometry corresponding to the A-DNA fibre model is not energetically favourable for the purine-pyrimidine sequences, which prefer small roll angle values when the slide has a large negative value as in A-DNA. However positive roll with large negative slide is allowed for GG, GA, AG and the pyrimidine-purine steps. This is consistent with the observed geometries of various steps in A-DNA crystals. Thus the general features of the basepair doublets predicted from these theoretical studies agree very well with the results from crystal structure analysis. However, since most sequences show an overall preference for B-type doublet geometry, the B----A transition for random sequence DNA cannot be explained on the basis of basepair stacking interactions.

Base Composition

Local variability and base sequence effects in DNA crystal structures.

The importance and usefulness of local doublet parameters in understanding sequence dependent effects has been described for A- and B-DNA oligonucleotide crystal structures. Each of the two sets of local parameters described by us in the NUPARM algorithm, namely the local doublet parameters, calculated with reference to the mean z-axis, and the local helical parameters, calculated with reference to the local helix axis, is sufficient to describe the oligonucleotide structures, with the local helical parameters giving a slightly magnified picture of the variations in the structures. The values of local doublet parameters calculated by NUPARM algorithm are similar to those calculated by NEWHELIX90 program, only if the oligonucleotide fragment is not too distorted. The mean values obtained using all the available data for B-DNA crystals are not significantly different from those obtained when a limited data set is used, consisting only of structures with a data resolution of better than 2.4 A and without any bound drug molecule. Thus the variation observed in the oligonucleotide crystals appears to be independent of the quality of their crystallinity. No strong correlation is seen between any pair of local doublet parameters but the local helical parameters are interrelated by geometric relationships. An interesting feature that emerges from this analysis is that the local rise along the z-axis is highly correlated with the difference in the buckle values of the two basepairs in the doublet, as suggested earlier for the dodecamer structures (Bansal and Bhattacharyya, in Structure & Methods: DNA & RNA, Vol. 3 (Eds., R.H. Sarma and M.H. Sarma), pp. 139-153 (1990)). In fact the local rise values become almost constant for both A- and B-forms, if a correction is applied for the buckling of the basepairs. In B-DNA the AA, AT, TA and GA basepair sequences generally have a smaller local rise (3.25 A) compared to the other sequences (3.4 A) and this seems to be an intrinsic feature of basepair stacking interaction and not related to any other local doublet parameter. The roll angles in B-DNA oligonucleotides have small values (less than +/- 8 degrees), while mean local twist varies from 24 degrees to 45 degrees. The CA/TG doublet sequences show two types of preferred geometries, one with positive roll, small positive slide and reduced twist and another with negative roll, large positive slide and increased twist.(ABSTRACT TRUNCATED AT 400 WORDS)

Algorithms

The tissue diagnosis of metabolic bone disease. Role of histomorphometry.

Those aspects of bone histology and histomorphometry that need to be understood by clinicians to put histomorphometry into proper perspective are presented. An increasingly important component of the histologic diagnosis of the metabolic diseases of bone is the quantification of the various histologic features as seen in undecalcified bone biopsies. This article is intended to review some of the histologic features of the metabolic diseases of bone and their corresponding histomorphometric parameters as they are used in diagnosis. The methods of obtaining both direct and derived values as well as the reporting of the results are presented in detail.

Biopsy

Molecular mechanics studies on poly(purine).poly(pyrimidine) sequences in DNA: polymorphism and local variability.

Energy minimization has been carried out on three poly(purine).poly(pyrimidine) sequences--d(G)10.d(C)10, d(A)10.d(T)10, and d(AG)5.d(CT)5--using the molecular mechanics program AMBER (Assisted Model Building and Energy Refinement). In order to extensively scan the conformational space available, five different helical models were studied, three of them being right-handed helices while the other two were left helical. For all three sequences the right-handed A- and B-type helices are energetically slightly preferred over the left helices, but the energy difference between the various right-handed helices is only marginal. A detailed analysis has been carried out to characterize the local structural variability in the refined structures, both in terms of torsion angles as well as other parameters such as base-pair tilt, wedge roll, and wedge tilt, etc. All three sequences exhibit similar structural features for a particular form, but both the forms A and B show significant deviations from fiber models. In particular, the A-form structures have higher unit rise (2.7 A), and lower unit twist (31 degrees) and base-pair tilt (12 degrees), compared to the fiber model, which has corresponding values of 2.56 A, 32.7 degrees, and 20 degrees, respectively. All these changes indicate that the refined models are closer to the A-form structure observed in crystals of oligonucleotides. In the refined B-for models, the helical parameters are close to the fiber B-form, although the torsion angles show considerable variations. None of the three sequences examined, including the d(A)n.d(T)n sequence, show any pronounced curvature for the B-form structure.

Base Sequence