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Biomedical subjects

P K Saha

Publications and source records attributed to P K Saha.

At least 19 recordsLinked to original sources

Sigma-nucleus potential in A=28.

We have studied the (pi(-),K+) reaction on a silicon target to investigate the sigma-nucleus potential. The inclusive spectrum was measured at a beam momentum of 1.2 GeV/c with an energy resolution of 3.3 MeV (FWHM) by employing the superconducting kaon spectrometer system. The spectrum was compared with theoretical calculations within the framework of the distorted-wave impulse approximation, which demonstrates that a strongly repulsive sigma-nucleus potential with a nonzero size of the imaginary part reproduces the observed spectrum.

Journal Article↗

Observation of a (6)(LambdaLambda)He double hypernucleus.

A double-hyperfragment event has been found in a hybrid-emulsion experiment. It is identified uniquely as the sequential decay of ( 6)(LambdaLambda)He emitted from a Xi(-) hyperon nuclear capture at rest. The mass of ( 6)(LambdaLambda)He and the Lambda-Lambda interaction energy DeltaB(LambdaLambda) have been measured for the first time devoid of the ambiguities due to the possibilities of excited states. The value of DeltaB(LambdaLambda) is 1.01+/-0.20(+0.18)(-0.11) MeV. This demonstrates that the Lambda-Lambda interaction is weakly attractive.

Journal Article↗

Artery-vein separation via MRA--an image processing approach.

This paper presents a near-automatic process for separating vessels from background and other clutter as well as for separating arteries and veins in contrast-enhanced magnetic resonance angiographic (CE-MRA) image data, and an optimal method for three-dimensional visualization of vascular structures. The separation process utilizes fuzzy connected object delineation principles and algorithms. The first step of this separation process is the segmentation of the entire vessel structure from the background and other clutter via absolute fuzzy connectedness. The second step is to separate artery from vein within this entire vessel structure via iterative relative fuzzy connectedness. After seed voxels are specified inside artery and vein in the CE-MRA image, the small regions of the bigger aspects of artery and vein are separated in the initial iterations, and further detailed aspects of artery and vein are included in later iterations. At each iteration, the artery and vein compete among themselves to grab membership of each voxel in the vessel structure based on the relative strength of connectedness of the voxel in the artery and vein. This approach has been implemented in a software package for routine use in a clinical setting and tested on 133 CE-MRA studies of the pelvic region and two studies of the carotid system from six different hospitals. In all studies, unified parameter settings produced correct artery-vein separation. When compared with manual segmentation/separation, our algorithms were able to separate higher order branches, and therefore produced vastly more details in the segmented vascular structure. The total operator and computer time taken per study is on the average about 4.5 min. To date, this technique seems to be the only image processing approach that can be routinely applied for artery and vein separation.

Algorithms↗

Breast tissue density quantification via digitized mammograms.

Studies reported in the literature indicate that breast cancer risk is associated with mammographic densities. An objective, repeatable, and a quantitative measure of risk derived from mammographic densities will be of considerable use in recommending alternative screening paradigms and/or preventive measures. However, image processing efforts toward this goal seem to be sparse in the literature, and automatic and efficient methods do not seem to exist. In this paper, we describe and validate an automatic and reproducible method to segment dense tissue regions from fat within breasts from digitized mammograms using scale-based fuzzy connectivity methods. Different measures for characterizing mammographic density are computed from the segmented regions and their robustness in terms of their linear correlation across two different projections--cranio-caudal and medio-lateral-oblique--are studied. The accuracy of the method is studied by computing the area of mismatch of segmented dense regions using the proposed method and using manual outlining. A comparison between the mammographic density parameter taking into account the original intensities and that just considering the segmented area indicates that the former may have some advantages over the latter.

Algorithms↗

Scale-based diffusive image filtering preserving boundary sharpness and fine structures.

Image acquisition techniques often suffer from low signal-to-noise ratio (SNR) and/or contrast-to-noise ratio (CNR). Although many acquisition techniques are available to minimize these, post acquisition filtering is a major off-line image processing technique commonly used to improve the SNR and CNR. A major drawback of filtering is that it often diffuses/blurs important structures along with noise. In this paper, we introduce two scale-based filtering methods that use local structure size or "object scale" information to arrest smoothing around fine structures and across even low-gradient boundaries. The first of these methods uses a weighted average over a scale-dependent neighborhood while the other employs scale-dependent diffusion conductance to perform filtering. Both methods adaptively modify the degree of filtering at any image location depending on local object scale. Object scale allows us to accurately use a restricted homogeneity parameter for filtering in regions with fine details and in the vicinity of boundaries while a generous parameter in the interiors of homogeneous regions. Qualitative experiments based on both phantoms and patient magnetic resonance images show significant improvements using the scale-based methods over the extant anisotropic diffusive filtering method in preserving fine details and sharpness of object boundaries. Quantitative analyses utilizing 25 phantom images generated under a range of conditions of blurring, noise, and background variation confirm the superiority of the new scale-based approaches.

Algorithms↗

Digital topological analysis of in vivo magnetic resonance microimages of trabecular bone reveals structural implications of osteoporosis.

Osteoporosis is a disease characterized by bone volume loss and architectural deterioration. The majority of work aimed at evaluating the structural implications of the disease has been performed based on stereologic analysis of histomorphometric sections. Only recently noninvasive imaging methods have emerged that provide sufficient resolution to resolve individual trabeculae. In this article, we apply digital topological analysis (DTA) to magnetic resonance microimages (mu-MRI) of the radius obtained at 137 x 137 x 350 microm3 voxel size in a cohort of 79 women of widely varying bone mineral density (BMD) and vertebral deformity status. DTA is a new method that allows unambiguous determination of the three-dimensional (3D) topology of each voxel in a trabecular bone network. The analysis involves generation of a bone volume fraction map, which is subjected to subvoxel processing to alleviate partial volume blurring, followed by thresholding and skeletonization. The skeletonized images contain only surfaces, profiles, curves, and their mutual junctions as the remnants of trabecular plates and rods after skeletonization. DTA parameters were compared with integral BMD in the lumbar spine and femur as well as MR-derived bone volume fraction (BV/TV). Vertebral deformities were determined based on sagittal MRIs of the spine with a semiautomatic method and the number of deformities counted after threshold setting. DTA structural indices were found the strongest discriminators of subjects with deformities from those without deformities. Subjects with deformities (n = 29) had lower topological surface (SURF) density (p < 0.0005) and surface-to-curve ratio (SCR; a measure of the ratio of platelike to rodlike trabeculae; p < 0.0005) than those without. Profile interior (PI) density, a measure of intact trabecular rods, was also lower in the deformity group (p < 0.0001). These data provide the first in vivo evidence for the structural implications inherent in postmenopausal osteoporosis accompanying bone loss, that is, the conversion of trabecular plates to rods and disruption of rods due to repeated osteoclastic resorption.

Adult↗

Topological analysis of trabecular bone MR images.

Recently, imaging techniques have become available which permit nondestructive analysis of the three-dimensional (3-D) architecture of trabecular bone (TB), which forms a network of interconnected plates and rods. Most osteoporotic fractures occur at locations rich in TB, which has spurred the search for architectural parameters as determinants of bone strength. In this paper, we present a new approach to quantitative characterization of the 3-D microarchitecture of TB, based on digital topology. The method classifies each voxel of the 3-D structure based on the connectivity information of neighboring voxels. Following conversion of the 3-D digital image to a skeletonized surface representation containing only one-dimensional (1-D) and two-dimensional (2-D) structures, each voxel is classified as a curve, surface, or junction. The method has been validated by means of synthesized images and has subsequently been applied to TB images from the human wrist. The topological parameters were found to predict Young's modulus (YM) for uniaxial loading, specifically, the surface-to-curve ratio was found to be the single strongest predictor of YM (r2 = 0.69). Finally, the method has been applied to TB images from a group of patients showing very large variations in topological parameters that parallel much smaller changes in bone volume fraction (BVF).

Aged↗

Morphological and cytoskeletal changes caused by non-membrane damaging cytotoxin of Vibrio cholerae on int 407 and HeLa cells.

Vibrio cholerae produces a non-membrane damaging cytotoxin (NMDCY), also known as cell rounding factor, which causes rapid rounding of cultured cells like HeLa, CHO and Vero and reportedly elicits enterotoxic activity in the rabbit ileal loop assay. Pursuing the concept that NMDCY might be an accessory factor contributing to the diarrhea caused by V. cholerae, we investigated the effect of NMDCY on Int 407 (intestinal cell line) and HeLa (non-intestinal cell line) cells using light, fluorescent and electron microscopy to gain insight into the cellular response evoked by NMDCY. Binding assays showed that NMDCY has affinity for both Int 407 and HeLa cells. Changes in the internal organelles and cytoskeletal structures of the cell lines were documented indicating changes in the secretory and metabolic function of the toxin-treated cells. Toxin-treated cells visualized under the electron microscope revealed retraction of cell body, formation of blebs on cell surface, changes in mitochondria having dilated and rarefied matrix and an extensively developed Golgi apparatus, endoplasmic reticulum and lysosomes compared to those in normal cells. Immunofluorescence study showed restructuring of microfilament network represented by actin, filamin and vinculin, as also of the microtubular component, tubulin and the intermediate filament, vimentin. Immunogold study further revealed that the toxin is internalized even within the nucleus. Moreover, a rise in the intracellular calcium level of the NMDCY-treated cells leads us to hypothesize that a cascade of events results in the final impairment of the cell machinery.

Calcium↗

Response of wild-type mutants of Vibrio cholerae O1 possessing different combinations of virulence genes in the ligated rabbit ileal loop and in Ussing chambers: evidence for the presence of additional secretogen.

Five wild-type mutant strains of Vibrio cholerae serogroup O1 that lacked the CTX virulence cassette, or contained a natural deletion of a virulence gene within the CTX virulence cassette, or possessed an additional virulence gene, along with a prototype toxigenic strain representing the El Tor classical biotypes were examined by in-vivo and in-vitro methods to determine their enterotoxic potential. The ability of whole cells and culture supernates of the strains to cause fluid accumulation in the rabbit ileal loop model revealed a pattern consistent with the presence of the various virulence gene(s), with those possessing the intact CTX virulence cassette being the most secretogenic. Culture supernates of strains without the CTX virulence cassette or the strain with an incomplete cassette were also able to evoke mild to moderate fluid accumulation in the rabbit ileal loop. Of the various media used, AKI and brain heart infusion broth appeared to support the production of a hitherto unknown secretogenic factor, because culture supernates of the non-toxigenic V. cholerae O1 strains showed higher fluid accumulation ratios when grown in these media than in the others. To confirm that the fluid accumulation elicited by these strains in the ileal loop was due to enterotoxin activity, the effect of supernate of the strains was examined in rabbit small intestine mounted on Ussing chambers. Increases in short circuit current and tissue conductance, as compared with the medium control, were observed even with the strains that did not possess the CTX virulence cassette, confirming their ability to disrupt the function of intestinal tissue. From these studies, it was concluded that strains of V. cholerae O1 devoid of the CTX virulence cassette were still able to elicit a secretory response in the ileal loop and displayed enterotoxic activity in an in-vitro experimental model.

Animals↗

Evaluation of serum creatine kinase as a diagnostic marker for tubal pregnancy.

Forty women comprising of 20 each in study and control groups had their serum creatine kinase (CK) estimated. The study subjects were proven cases of ectopic pregnancy and the controls were either medical termination of pregnancy seekers or antenatal cases matched for gestational age and with a confirmed intrauterine pregnancy. Total serum creatine kinase levels were found to be significantly higher in the study group i.e. 34.15 +/- 1.17 IU/L compared to the controls 18.72 +/- 1.25 (p < 0.001). Creatine kinase is estimated routinely in cardiac patients even on an emergency basis. Our findings indicate that when in doubt the test could be used as a marker for the diagnosis of ectopic pregnancy.

Biomarkers↗

Clonal analysis of non-toxigenic Vibrio cholerae O1 associated with an outbreak of cholera.

We examined the clonal relationships among eight clinical isolates of non-toxigenic (NT) V. cholerae O1 associated with a cluster of cases of cholera in Warangal, Andhra Pradesh in south India and compared their relatedness to toxigenic O1 strains of classical and E1Tor biotypes and with O139 Bengal strains of V. cholerae by pulsed-field gel electrophoresis (PFGE). Phylogentic analysis of the NotI restriction fragment length polymorphism showed that all the NT. V. cholerae O1 strains formed a tight cluster with more than 80 per cent similarity. Interestingly, the NT V. cholerae O1 cluster was more closely related to V. cholerae O139 than to classical and E1Tor biotypes of V. cholerae O1 indicating closer genetic relationships between NT V. cholerae 01 and O139 Bengal strains that were isolated during the same time-frame.

Bacterial Typing Techniques↗

Characterization of non-membrane-damaging cytotoxin of non-toxigenic Vibrio cholerae O1 and its relevance to disease.

The non-membrane-damaging cytotoxin which causes dramatic cell rounding of cultured HeLa cells was purified to homogeneity from a clinical strain (WO5) of non-toxigenic Vibrio cholerae O1 Inaba belonging to the E1 Tor biotype. The purified protein has a denatured molecular weight of 35 kDa and a native molecular weight of approximately 37 kDa indicating the monomeric nature of the protein. The 15 N-terminal amino acid sequence of non-membrane-damaging cytotoxin showed complete homology to the hemagglutinin protease previously purified and characterized from V. cholerae O1. Purified non-membrane-damaging cytotoxin from V. cholerae O1 was immunologically and biochemically identical to that previously purified from V. cholerae O26. Non-membrane-damaging cytotoxin was found to be enterotoxic in rabbit ileal loop assay inducing accumulation of non-hemorrhagic fluid at 100 micrograms and elicited a concentration dependent increase in short circuit current and tissue conductance of rabbit ileal mucosa mounted on Ussing chambers. A significant serum immunoglobulin G response against non-membrane-damaging cytotoxin was elicited by patients infected with V. cholerae O139 but not with V. cholerae O1. These properties make non-membrane-damaging cytotoxin a potential virulence factor of V. cholerae which should be taken into consideration while making live, attenuated recombinant vaccine strains against cholera.

Animals↗

Production of monoclonal antibodies to the non-membrane-damaging cytotoxin (NMDCY) purified from Vibrio cholerae O26 and distribution of NMDCY among strains of Vibrio cholerae and other enteric bacteria determined by monoclonal-polyclonal sandwich enzyme-linked immunosorbent assay.

The distribution of a newly described secretogenic non-membrane-damaging cytotoxin (NMDCY) among strains of Vibrio cholerae and other enteric bacteria was determined. To accomplish this, monoclonal antibodies against NMDCY were prepared and a sandwich monoclonal-polyclonal enzyme-linked immunosorbent assay (ELISA) was developed. By the sandwich ELISA, it was determined that 55.6% of the 412 strains of V. cholerae examined produced NMDCY at varying concentrations while 76, 37.9, and 15.6% of the clinical strains of Vibrio parahaemolyticus, Aeromonas spp., and Shigella spp., respectively, produced NMDCY. Because of its enterotoxigenic potential and based on its widespread distribution among strains of V. cholerae, we believe that NMDCY may constitute an important virulence determinant in the cascade of events which enable the organism to precipitate the disease.

Animals↗

Purification and characterization of an extracellular secretogenic non-membrane-damaging cytotoxin produced by clinical strains of Vibrio cholerae non-O1.

Some clinical strains of Vibrio cholerae non-O1 produce an extracellular factor that evokes a rapid and dramatic cytotoxic response which manifests as cell rounding of Chinese hamster ovary (CHO) and HeLa cells without accompanying membrane damage. This study was performed to establish the identity of the non-membrane-damaging cytotoxin (NMDCY), which was not inhibited by antitoxins against cholera toxin, heat-labile toxin of enterotoxigenic Escherichia coli, El Tor hemolysin, Shiga-like toxin I, and Shiga-like toxin II, indicating that NMDCY did not bear an apparent immunological relationship with the above toxins and hemolysin. Brain heart infusion broth and AKI medium supported the maximal production of NMDCY; culture supernatant of AKI medium was found to be free of hemolysin activity, whereas in brain heart infusion broth hemolysin was coproduced with NMDCY. Maximal production of NMDCY in AKI medium was observed at 37 degrees C under shaking conditions with the pH of the medium adjusted to 8.5. NMDCY was purified to homogeneity by a three-step purification procedure which increased the specific activity of the cytotoxin by 1.7 X 10(5)-fold. The denatured molecular weight of the purified toxin was 35,000, and the cytotoxin was heat labile and sensitive to trypsin. Purification of the cytotoxin revealed an enterotoxic activity as reflected by its ability to accumulate fluid in the rabbit ileal loop. Both the cytotoxic and enterotoxic activities of NMDCY could be inhibited or neutralized by antiserum raised against purified cytotoxin but not by preimmune serum. Immunodiffusion test between purified NMDCY and antiserum gave a single well-defined precipitin band which showed reactions of complete identity, while, in an immunoblot assay, a well-defined single band was observed in the 35-kDa region. Our results indicate that the cytotoxic and enterotoxic activities expressed by NMDCY appear to contribute to the pathogenesis of the disease associated with V. cholerae non-O1 strains which produce this cytotoxin.

Animals↗

Nontoxigenic Vibrio cholerae 01 serotype Inaba biotype El Tor associated with a cluster of cases of cholera in southern India.

Thirteen strains of Vibrio cholerae 01 belonging to the Inaba serotype El Tor biotype isolated from patients during an outbreak of cholera in the town of Warangal in southern India were found to be nontoxigenic (NT), since they did not produce cholera toxin or hybridize with DNA probes specific for cholera toxin, Zot, or Ace. The unheated and heated culture supernatants of the NT V. cholerae 01 evoked a rapid cell-rounding effect when introduced on confluent layers of CHO and HeLa cells which could not be inhibited by antiserum against known toxins. Culture supernatants of two representative NT V. cholerae 01 strains caused an increase in short-circuit current in rabbit ileal tissue mounted on an Ussing chamber, and the pattern of increase in short-circuit current was consistent with the presence of a quickly acting toxin like stable toxin. None of the strains of NT V. cholerae 01 hybridized with a DNA probe specific for the heat-stable enterotoxin of V. cholerae non-01, nor did the factor produced by NT V. cholerae 01 resemble the recently described heat-stable enterotoxin produced by enteroaggregative Escherichia coli as determine by a PCR assay. To our knowledge, this is the first report of NT V. cholerae 01 being associated with a cluster of cases of cholera, and it appears that a clone of NT V. cholerae 01 has the potential to cause localized outbreaks of cholera.

Animals↗

Comparative analysis of factors promoting optimal production of cholera toxin by Vibrio cholerae O1 (classical & E1Tor biotypes) & O139.

Various culture media [AKI, Brain heart infusion broth (BHI), Casamino acid-yeast extract broth (CAYE), Casamino acid-yeast extract broth supplemented with 90 micrograms/ml of lincomycin (CAYE-L), Tryptic soy broth (TSB) and Yeast extract peptone (YEP)], cultural conditions (stationary and shaking) and incubation temperatures (30 degrees C and 37 degrees C) were evaluated to determine optimal conditions for production of cholera toxin (CT) by different biotypes (classical and E1Tor) and serogroups (O1 and O139) of V. cholerae. It was found that V. cholerae O1 E1Tor grown in CAYE-L and incubated at 30 degrees C with constant shaking was optimal for production of CT, while for the classical biotype and for the O139 serogroup, CT was maximally produced when grown in YEP and incubated at 30 degrees C in a shaker. Temperature appeared to be a prominent factor affecting the production of CT by the O1 E1Tor biotype when the media used were AKI, CAYE-L and YEP and also for the classical biotype when the media used were the AKI, BHI, CAYE and YEP. In the case of the O1 E1Tor biotype, CAYE-L was the best medium for CT production whereas for the classical biotype, CAYE-L was a poor medium as far as CT production was concerned. Irrespective of the media used, 30 degrees C shake culture condition seemed to be more favourable for supporting CT production except in CAYE medium for the O1 E1Tor biotype where incubation at 37 degrees C in a shaker was as good as incubation at 30 degrees C.

Cholera Toxin↗

Distribution and virulence of Vibrio cholerae belonging to serogroups other than O1 and O139: a nationwide survey.

The distribution and virulence of Vibrio cholerae serogroups other than O1 and O139 in India before, during and after the advent of O139 serogroup was investigated. A total of 68 strains belonging to 31 different 'O' serogroups were identified during the study period. With the exception of O53, there was no spatial or temporal clustering of any particular non-O1 non-O139 serogroup at any given place. Two of the 68 strains examined produced cholera toxin (CT) which could only be partially absorbed with anti-CT immunoglobulin G. Tissue culture assay revealed that some of the non-O1 non-O139 strains produced factors which evoked either a cell rounding or cell elongation response depending upon the medium used. This study indicates that serogroups other than O1 and O139 should also be continuously monitored.

Animals↗