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S K Sahni

Publications and source records attributed to S K Sahni.

15 recordsLinked to original sources
PubMed2001

An algorithm for automatic, computed-tomography-based source localization after prostate implant.

Permanent implant of the prostate using I-125 and Pd-103 seeds is a popular choice of treatment for early-stage prostate cancer in the United States. Evaluation of the quality of the implant is best based on the calculated dose distribution from postimplant computed tomography (CT) images. This task, however, has been time-consuming and inaccurate. We have developed an algorithm for automatic source localization from postimplant CT images. The only requirement of this algorithm is knowledge of the number of seeds present in the prostate, thus minimizing the need for human intervention. The algorithm processes volumetric CT data from the patient, and pixels of higher CT numbers are categorized into classes of definite and potential source pixels. A multithresholding technique is used to further determine the number of seeds and their precise locations in the CT volume data. A graphic user interface was developed to facilitate operator review of and intervention in the calculation and the results of the algorithm. This algorithm was tested on two phantoms containing nonradioactive seeds, one with 20 seeds in discrete locations and another with 100 seeds with small distances between seeds. The tests showed that the algorithm was able to identify the seed locations to within 1 mm of their physical locations for discrete seed locations. It was further able to separate seeds at close proximity to each other while maintaining an average seed localization error of less than 2 mm, with no operator intervention required.

Algorithms↗
PubMed1999

Involvement of protein kinase C in Rickettsia rickettsii-induced transcriptional activation of the host endothelial cell.

Our laboratory has reported on a biphasic pattern of nuclear factor kappaB (NF-kappaB) activation in cultured human umbilical vein endothelial cells during infection with Rickettsia rickettsii, an obligate, intracellular bacterium, and the etiologic agent of Rocky Mountain spotted fever. Transcriptional activation of the tissue factor (TF) gene during this infection has been shown to involve NF-kappaB. To further understand the signal transduction events underlying these phenomena, we studied the role of protein kinase C (PKC), a ubiquitous family of phospholipid-dependent enzymes implicated in the regulation of a variety of cell signaling pathways. Two inhibitors of PKC, namely, bisindolylmaleimide I hydrochloride (BM-1) and calphostin C, which exhibit different inhibitory properties towards various isozymes of PKC, were used. Infection of cells with R. rickettsii in the presence of BM-1 (50 nM) did not significantly affect NF-kappaB, whereas calphostin C (2.5 microM) completely blocked the early phase of NF-kappaB activation. The late, sustained phase also was not affected by treatment with BM-1. Downregulation of phorbol ester-sensitive PKCs by long-term treatment with phorbol 12-myristate 13-acetate (PMA) did not inhibit NF-kappaB activation. Likewise, this downregulation had no effect on induction of TF activity. The activity of TF was, however, sensitive to BM-1 and calphostin C, whereas expression of TF mRNA was inhibited only by calphostin C. Overall, these results suggest the lack of involvement of classical PKC pathways in R. rickettsii-induced NF-kappaB activation but the possible involvement of a non-PMA-responsive PKC isoform in the posttranscriptional control of TF expression.

Cells, Cultured↗
PubMed1998

NF-kappa B-dependent inhibition of apoptosis is essential for host cellsurvival during Rickettsia rickettsii infection.

The possibility that bacteria may have evolved strategies to overcome host cell apoptosis was explored by using Rickettsia rickettsii, an obligate intracellular Gram-negative bacteria that is the etiologic agent of Rocky Mountain spotted fever. The vascular endothelial cell, the primary target cell during in vivo infection, exhibits no evidence of apoptosis during natural infection and is maintained for a sufficient time to allow replication and cell-to-cell spread prior to eventual death due to necrotic damage. Prior work in our laboratory demonstrated that R. rickettsii infection activates the transcription factor NF-kappa B and alters expression of several genes under its control. However, when R. rickettsii-induced activation of NF-kappa B was inhibited, apoptosis of infected but not uninfected endothelial cells rapidly ensued. In addition, human embryonic fibroblasts stably transfected with a superrepressor mutant inhibitory subunit Ikappa B that rendered NF-kappa B inactivatable also underwent apoptosis when infected, whereas infected wild-type human embryonic fibroblasts survived. R. rickettsii, therefore, appeared to inhibit host cell apoptosis via a mechanism dependent on NF-kappa B activation. Apoptotic nuclear changes correlated with presence of intracellular organisms and thus this previously unrecognized proapoptotic signal, masked by concomitant NF-kappa B activation, likely required intracellular infection. Our studies demonstrate that a bacterial organism can exert an antiapoptotic effect, thus modulating the host cell's apoptotic response to its own advantage by potentially allowing the host cell to remain as a site of infection.

Apoptosis↗
PubMed1998

Proteasome-independent activation of nuclear factor kappaB in cytoplasmic extracts from human endothelial cells by Rickettsia rickettsii.

Interaction of many infectious agents with eukaryotic host cells is known to cause activation of the ubiquitous transcription factor nuclear factor kappaB (NF-kappaB) (U. Siebenlist, G. Franzoso, and K. Brown, Annu. Rev. Cell Biol. 10:405-455, 1994). Recently, we reported a biphasic pattern of NF-kappaB activation in cultured human umbilical vein endothelial cells consequent to infection with Rickettsia rickettsii, an obligate intracellular gram-negative bacterium and the etiologic agent of Rocky Mountain spotted fever (L. A. Sporn, S. K. Sahni, N. B. Lerner, V. J. Marder, D. J. Silverman, L. C. Turpin, and A. L. Schwab, Infect. Immun. 65:2786-2791, 1997). In the present study, we describe activation of NF-kappaB in a cell-free system, accomplished by addition of partially purified R. rickettsii to endothelial cell cytoplasmic extracts. This activation was rapid, reaching maximal levels at 60 min, and was dependent on the number of R. rickettsii organisms added. Antibody supershift assays using monospecific antisera against NF-kappaB subunits (p50 and p65) confirmed the authenticity of the gel-shifted complexes and identified both p50-p50 homodimers and p50-p65 heterodimers as constituents of the activated NF-kappaB pool. Activation occurred independently of the presence of endothelial cell membranes and was not inhibited by removal of the endothelial cell proteasome. Lack of involvement of the proteasome was further confirmed in assays using the peptide-aldehyde proteasome inhibitor MG 132. Activation was not ATP dependent since no change in activation resulted from addition of an excess of the unhydrolyzable ATP analog ATPgammaS, supplementation with exogenous ATP, or hydrolysis of endogenous ATP with ATPase. Furthermore, Western blot analysis before and after in vitro activation failed to demonstrate phosphorylation of serine 32 or degradation of the cytoplasmic pool of IkappaB alpha. This lack of IkappaB alpha involvement was supported by the finding that R. rickettsii can induce NF-kappaB activation in cytoplasmic extracts prepared from T24 bladder carcinoma cells and human embryo fibroblasts stably transfected with a superrepressor phosphorylation mutant of IkappaB alpha, rendering NF-kappaB inactivatable by many known signals. Thus, evidence is provided for a potentially novel NF-kappaB activation pathway wherein R. rickettsii may interact with and activate host cell transcriptional machinery independently of the involvement of the proteasome or known signal transduction pathways.

Cells, Cultured↗
PubMed1997

Rickettsia rickettsii infection of cultured human endothelial cells induces NF-kappaB activation.

Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever, is an obligate intracellular bacterial organism that infects primarily the vascular endothelial cells (EC). A component of the EC response to infection is transcriptional activation, which may contribute to the thrombotic and inflammatory consequences of disease. In this study, we explore R. rickettsii-induced activation of the nuclear factor-kappaB/Rel (NF-kappaB) family of transcription factors involved in early transcriptional responses to injurious stimuli. Two NF-kappaB species were activated by infection and reacted with a double-stranded oligonucleotide probe corresponding to the kappaB binding domain of the murine kappa light-chain gene enhancer. Gel supershift analysis demonstrated the reactivity of these complexes with antibodies against p65 and p50, and the induced species were tentatively identified as p50-p50 homodimers and p50-p65 heterodimers. Semiquantitative reverse transcription-PCR analysis revealed dramatic increases in the steady-state levels of mRNA coding for the inhibitory subunit of NF-kappaB (IkappaB alpha), transcription of which is enhanced by the binding of NF-kappaB within the IkappaB alpha promoter region. NF-kappaB activation was first detected 1.5 h following infection and was biphasic, with an early peak of activation at approximately 3 h, a return to baseline levels at 14 h, and even higher levels of activation at 24 h. It is likely that NF-kappaB activation requires cellular uptake of R. rickettsii, since treatment of EC with cytochalasin B during infection to block entry inhibited activation by only 70% at 3 h. R. rickettsii-induced activation of NF-kappaB may be an important controlling factor in the transcriptional responses of EC to infection with this obligate intracellular organism.

Cells, Cultured↗
PubMed1995

Amperometric thin film biosensors based on glucose dehydrogenase and Toluidine Blue O as catalyst for NADH electrooxidation.

Amperometric glucose sensors were constructed based on solid graphite electrodes, surface-modified with NAD+ dependent glucose dehydrogenase (GDH), Toluidine Blue O (TBO), and protective ionic polymers. The electrocatalytic oxidation of NADH was evaluated from cyclic voltammetry with TBO dissolved, adsorbed, and electrostatically or covalently bound to polymers. The NADH and glucose sensors constructed were investigated and operated at 0 mV vs. Ag/AgCl using single potential step chronoamperometry. The operational stability of the glucose sensors was limited by leakage of NAD+. A glucose sensitivity much higher than carbon paste electrode was found. A sensitivity as high as 25 microA cm-2 mM-1 was achieved.

Biosensing Techniques↗
PubMed1993

Repeated cocaine administration does not affect 5-HT receptor subtypes (5-HT1A, 5-HT2) in several rat brain regions.

In order to examine whether cocaine-induced behavioral sensitization is modulated by changes in serotonin receptor subtypes, we measured the binding of [3H]8-hydroxy-2-(di-n-propylamino)tetralin ([3H]8-OH-DPAT) to 5-HT1A receptors and of [3H]-ketanserin to 5-HT2 receptors in various brain regions of cocaine-treated and saline-treated (control) rats. As previously reported, repeated administration of cocaine resulted in behavioral sensitization. Stereotypic scores with the cocaine challenge were significantly (P < 0.05) higher in cocaine-pretreated animals than in the saline-pretreated group. Neither acute nor chronic cocaine administration significantly altered the number (Bmax) or the affinity (KD) of either [3H]8-OH-DPAT or [3H]ketanserin binding sites in any of the brain regions examined. These results suggest that the enhanced functional sensitivity of 5-HT1A or 5-HT2 receptor subtypes seen with cocaine may be associated with alterations in processes distal to receptors rather than changes in the number or the affinity of the receptors.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1992

Alterations in host regulatory glycolytic enzymes during pathogenesis of primate malaria and following chloroquine therapy.

Hexokinase, phosphofructokinase and pyruvate kinase, the regulatory enzymes of the glycolytic sequence, showed progressive increases in their activities with the rise of parasitaemia in Plasmodium knowlesi-infected rhesus monkey serum and red blood cells. Chloroquine therapy cleared the parasitaemia in 72 h and brought the elevated levels of these enzymes back to almost normal in about 30 days.

Animals↗
PubMed1992

Studies on glycolytic enzymes of Plasmodium knowlesi, a simian malaria parasite, and infected host erythrocytes.

Almost all of the important enzymes of glycolytic sequence were studied in isolated schizonts of Plasmodium knowlesi as well as in normal and P. knowlesi parasitized rhesus red blood cells. Significant activities of all the glycolytic enzymes assayed were observed in cell-free schizonts and the levels of all were found to be considerably elevated in infected host erythrocytes, confirming that the consumption of glucose is dramatically increased in host red cells, consequent to malaria infection.

Animals↗
PubMed1991

Status of hepatic heme and heme oxygenase during Plasmodium yoelii nigeriensis infection in mice.

Plasmodium yoelii nigeriensis infection in albino mice caused a significant increase in hepatic heme level, the increase being concomitant with a rise in parasitemia. This elevated heme was found to be associated with all the subcellular fractions except the cytosol, where its content remained unaltered. Activity of heme oxygenase, the key enzyme responsible for catabolism of heme, also increased progressively with rise in parasitemia. Treatment of normal mice with cobalt chloride [60 mg (kg body wt)-1; subcutaneously] brought about a 150% increase in the level of heme oxygenase; similar treatment of infected mice at low parasitemia could induce the enzyme activity while at high parasitemia the enzymic activity remained unaltered as compared to untreated infected mice. In spite of an increased level of heme oxygenase in the cobalt-treated mice, the level of heme did not show any noticeable change. Oral administration of chloroquine [64 mg (kg body wt)-1 x 4 days] brought about a 56% reduction in the level of heme oxygenase of normal animals but there was no change in infected animals when compared with the corresponding untreated infected mice. However, the amount of chloroquine present in livers of normal and infected animals was not significantly different.

Administration, Oral↗
PubMed1988

Intraoperative electroneurography: management of ulnar neuropathy at the elbow.

At the elbow the ulnar nerve may be compressed either in the retrocondylar groove or at the cubital tunnel. Optimal surgical therapy should be directed at the specific site of involvement. Intraoperative electroneurography performed in conjunction with 19 ulnar nerve explorations helped localize the precise site of compression. Of the primary procedures, abnormality was at the retrocondylar groove in 9, cubital tunnel in 4, both locations in 3, and at an unusual distal point in 1; 12 anterior subcutaneous transpositions, 4 cubital tunnel releases, and 1 distal decompression resulted. Intraoperative studies helped identify residual compression in two patients undergoing reexploration. Although routine electrodiagnosis may localize an ulnar neuropathy to the elbow, reliably separating retrocondylar from cubital tunnel compression is more difficult. Preoperatively, percutaneous serial short increment studies were more accurate than simple "inching" in predicting the site of compression.

Diagnosis, Differential↗
PubMed1988

Entrapment neuropathy of the ulnar nerve at its point of exit from the flexor carpi ulnaris muscle.

The ulnar nerve normally enters the flexor carpi ulnaris (FCU) proximally and anteriorly between the humeral and ulnar heads of the muscle. After an intramuscular course of several centimeters, the nerve exits the FCU distally to lie in a tissue plane between the FCU and the flexor digitorum profundus (FDP). A patient with ulnar neuropathy studied by intraoperative electroneurography demonstrated major focal conduction block at the point where the nerve exited the FCU. A fivefold increase in amplitude and reversal of a dispersed, irregular compound muscle action potential to a more normal configuration occurred with stimulation just distal to the point of exit. There was no evidence by inspection, probing, or electroneurography of compression in the retrocondylar groove or at the cubital tunnel. In a series of 100 cadaver dissections, the intermuscular septum between FCU and FDP was thick and tough in several specimens. This septum may represent a site of ulnar nerve entrapment.

Electrodiagnosis↗
PubMedDate not supplied

Effect of anti-relapse antimalarial compound CDRI 80/53 and primaquine on hepatic mixed function oxidase system of rhesus monkey.

A potential anti-relapse antimalarial compound CDRI 80/53 [N1-(3-acetyl-4-5-dihydro-2-furanyl)-N4-(6-methoxy-8-quinolinyl)1,4- pentanediamine] at 7.5 mg/kg body weight and primaquine at 6.0 mg/kg body weight did not cause any significant change in the status of hepatic microsomal mixed function oxidase system of rhesus monkeys, when given orally for 7 days. Further, the extension of the treatment at the same dossier up to 21 days resulted in impairment of the different indices of the MFO system. The compound CDRI 80/53 inhibited cytochrome P-450, aminopyrine-N-demethylase, aniline and benzo(a)pyrene hydroxylases, cytochrome b5 and haem levels by 17, 11, 58, 0, 36 and 35% whereas the inhibition caused by primaquine was 34, 40, 72, 54, 39 and 38% respectively, establishing that the cytochrome P-450 dependent mono-oxygenase system of monkey liver was comparatively less suppressed by compound CDRI 80/53. The cessation of the compound/drug treatment resulted in almost complete reversal of all the MFO activities to normal in a period of about 6 weeks.

Aminoquinolines↗
PubMedDate not supplied

NADP-specific isocitrate dehydrogenase from the simian malaria parasite Plasmodium knowlesi: partial purification and characterization.

Cell-free schizonts of Plasmodium knowlesi, a simian malaria parasite, possess significant isocitrate dehydrogenase (IDH) activity, about 90% of which is contributed by the NADP-specific enzyme that is localized in the cytosolic fraction. The enzyme has been partially purified by affinity chromatography using Blue sepharose CL-6B. Although unstable in nature, it is stabilized by citrate and glycerol. Kinetic studies with DL-isocitrate and NADP yielded hyperbolic curves with Michaelis constants of 0.210 and 0.038 mM, respectively. Manganous or magnesium ions are essential for activity. The enzyme is thermosensitive, shows maximum activity at pH 8.0, and has a molecular mass of about 48.5 kDa. It is strongly inhibited by thiol-blocking agents but protected against them by thiol-providing agents. Cupric and argentic ions also have a marked inhibitory effect on its activity. The enzyme is significantly inhibited by chloroquine and oxytetracycline in vitro, but to a lesser degree by tetracycline.

Animals↗