Biomedical subjects
Publications and source records attributed to Himanshu Shah.
Structure-activity relationship study of bipiperidine amide 1 has identified the reverse bipiperidine amide 4a as a CC chemokine-3 (CCR3) receptor antagonist. Optimization of the structure-activity relationship of compound 4a has resulted in the identification of a CCR3 antagonist 4i as well as a CCR3 agonist 13.
Bipiperidine amide 1 has been identified as a CC chemokine receptor 3 (CCR3) antagonist. Optimization of its structure-activity relationship has resulted in the identification of cis (R,R)-4-[(3,4-dichlorophenyl)methyl]-3-hydroxymethyl-1'(6-quinolinylcarbonyl)-1,4'-bipiperidine 14n, which exhibits potent receptor affinity and inhibition of both calcium flux and eosinophil chemotaxis.
A 40-year-old male with alcoholic cirrhosis and portal hypertension presented with acute variceal hemorrhage. Abdominal CT scan and endoscopy revealed large gastric varices. The patient underwent a TIPS procedure. Portal venography demonstrated persistent filling of the large gastric varices with associated high-flow spontaneous splenorenal shunt. Because of the large size of the varices, a Simon-Nitinol filter was used in conjunction with multiple embolization coils to enable successful occlusion of the varices.
PURPOSE: To determine diagnostic accuracy of four-channel multi-detector row computed tomography (CT) in emergency room and inpatient populations suspected of having acute pulmonary embolism (PE) who prospectively underwent both CT and pulmonary arteriography (PA). MATERIALS AND METHODS: Patients referred for PA to assess suspected PE were eligible. Institutional review board approval and written informed consent were obtained. All patients underwent CT and PA within a 48-hour period. For CT, 4 x 2.5-mm collimation was used. Three readers independently evaluated each study for PE presence. PE status, vessel level, and lobar location were determined by means of majority rule, and interobserver agreement (kappa) was calculated for PE status, as assessed with each modality. Sensitivity and specificity of CT were calculated by using PA as the reference standard. Two radiologists later reviewed false-positive CT studies. RESULTS: The study group comprised 93 patients (median age, 56 years; range, 19-88 years). Sensitivity, specificity, and accuracy of CT were 100%, 89%, and 91%, respectively. kappa values were 0.71 and 0.83 for CT and PA, respectively, and were not significantly different between modalities. At PA, 18 patients (19%) had PE at 50 vessel levels (five main and/or interlobar, 24 segmental, and 21 subsegmental), 17 (94%) of which had PE at multiple sites. At CT, 26 patients (28%) had PE at 71 vessel levels (24 main and/or interlobar, 33 segmental, and 14 subsegmental). Twenty patients (77%) had PE at multiple sites. Review of eight false-positive CT studies showed an appearance highly suggestive of acute PE in three patients, chronic PE in one, and no PE in three; one study was inconclusive. CT better demonstrated large-level vessel involvement (P < .01), while PA better demonstrated small-level vessel involvement (P < .01). CONCLUSION: Multi-detector row CT has an accuracy of 91% in the depiction of suspected acute PE when conventional PA is used as the reference standard.
ADAM33 (a disintegrin and metalloproteinase) is an asthma susceptibility gene recently identified through a genetic study of asthmatic families (van Eerdewegh et al. (2002) Nature 418, 426-430). In order to characterize the catalytic properties of ADAM33, the metalloproteinase domain of human ADAM33 was expressed in Drosophila S2 cells and purified. The N-terminal sequence of the purified metalloproteinase was exclusively (204)EARR, indicating utilization of one of three furin recognition sites. Of many synthetic peptides tested as potential substrates, four peptides derived from beta-amyloid precursor protein (APP), Kit-ligand-1 (KL-1), tumor necrosis factor-related activation-induced cytokine, and insulin B chain were cleaved by ADAM33; mutation at the catalytic site, E346A, inactivated catalytic activity. Cleavage of APP occurred at His(14)/Gln(15), not at the alpha-secretase site and was inefficient (k(cat)/K(m) (1.6 +/- 0.3) x 10(2) m(-1) s(-1)). Cleavage of a juxtamembrane KL-1 peptide occurred at a site used physiologically with a similar efficiency. Mutagenesis of KL-1 peptide substrate indicated that the P3, P2, P1, and P3' residues were critical for activity. In a transfected cell-based sheddase assay, ADAM33 functioned as a negative regulator of APP shedding and mediated some constitutive shedding of KL-1, which was not regulated by phorbol 12-myristate 13-acetate activation. ADAM33 activity was sensitive to several hydroxamate inhibitors (IK682, K(i) = 23 +/- 7 nm) and to tissue inhibitors of metalloproteinase (TIMPs). Activity was inhibited moderately by TIMP-3 and TIMP-4 and weakly inhibited by TIMP-2 but not by TIMP-1, a profile distinct from other ADAMs. The identification of ADAM33 peptide substrates, cellular activity, and a distinct inhibitor profile provide the basis for further functional studies of ADAM33.
We compared the tissue mRNA prevalence and protein maturation of two splice variants of mouse ADAM33, a metalloprotease implicated in airway hyperresponsiveness. These variant cDNAs, designated 914 (alpha) and 906 (beta), encode membrane-bound forms that differ primarily in 26 residues (exon 17) between the cysteine-rich and epidermal growth factor-like domains. Proteins of approximately 120 and 103 kD, detectable by anti-ADAM33 antibodies, were expressed in 914-transfected HEK293 cells. The time-dependent appearance of the approximately 100-kD form and its inhibition by a peptidyl chloromethylketone, or the calcium ionophore, A23187, indicated that this was mature ADAM33, which was processed by a furin-like convertase. One form, approximately 110 kD, was detected in 906-transfected cell lysates. Trypsin and biotinylation treatment of transfected cells demonstrated that all of the mature approximately 100-kD, a minority of the approximately 120-kD pro-form, and none of the 906-expressed 110-kD form localized to the cell surface. The mature form was resistant to endoglycosidase H(f). The approximately 110-kD form was endoglycosidase H(f)-sensitive, indicating retention proximal to the trans-Golgi, consistent with a lack of maturation. Quantitation of transcripts demonstrated that those containing exon 17 predominate, whereas those lacking exon 17 are negligible in the mouse lung, although detectable at low levels in mouse testis, heart, and brain. Thus, potential dominant-negative effects exerted by the nonprocessed 906-encoded beta splice variant are unlikely to occur in mouse lung.
We examined transcript expression and post-transcriptional regulation of human ADAM33, a recently identified asthma gene. A detailed messenger RNA (mRNA) expression profile was obtained using Northern, reverse transcription polymerase chain reaction, and in situ hybridization analyses. ADAM33 mRNA was expressed significantly in smooth muscle-containing organs, minimally in immune organs and hematopoietic cells, and highly in repairing duodenal granulation tissue. Expression was seen in asthmatic subepithelial fibroblasts and smooth muscle but not in respiratory epithelium. In all tissues, transcripts of approximately 5 kb predominated over those of approximately 3.5 kb by 2- to 5-fold. The effect of the 3' untranslated region (UTR) on ADAM33 protein expression and maturation was examined. The presence of the 3'UTR in untagged full-length constructs promoted prodomain removal, detected as mature approximately 100 kD protein by ADAM33-reactive antibodies; in its absence, maturation was 2- to 3-fold less in HEK293 cells. His-tagged and untagged constructs lacking the 3'UTR demonstrated that lack of maturation was not a result of tag-mediated effects. Minimal maturation of ADAM33 occurred in primary lung and MRC5 fibroblasts following adenoviral-mediated expression of ADAM33 lacking the 3'UTR. In contrast, prodomain removal was observed with plasmids and adenovirus encoding only the pro- and catalytic domains. Thus, the 3'UTR of ADAM33 and domains downstream of the catalytic domain regulate potential ADAM33 activity. Mechanisms of regulation of ADAM33, distinct from closely related ADAMs, thus include mRNA localization and processing and protein maturation.
ADAM33 (a disintegrin and metalloprotease) was recently found to be a novel asthma susceptibility gene. Domain-specific antibodies were used to study its expression and processing. When the pro-domain and catalytic domain were expressed by a stable-transfected cell line, the pro-domain was removed by cleavage within a putative furin cleavage site. The catalytic domain was active in an alpha(2)-macroglobulin complex formation assay and mutation of the catalytic site glutamic acid (E346A) eliminated activity. In transient transfections using the full-length protein, a pro-form and mature form were detectable and alternate glycosylation was demonstrated at sites within the catalytic domain. ADAM33 was detected on the cell surface, with the majority of protein detected intracellularly. The E346A mutation had no significant effect on protein processing. Endogenous ADAM33 was detected in bronchus tissue, bronchial smooth muscle cells, and MRC-5 fibroblasts, consistent with a role in the pathophysiology of asthma.
Study of the CC chemokine receptor 3 (CCR3) has been limited to using radiolabeled agonist chemokines. A small molecule CCR3 antagonist, 2-[(6-amino-2-benzothiazolyl)thio]-N-[1-[(3,4-dichlorylphenyl)methyl]-4-piperidinyl]acetamide, Banyu (I), was tritiated and used for pharmacological studies. Banyu (I) has a K(d) of 5.0+/-0.4 and 4.3+/-1.8 nM on human CCR3 transfectants and eosinophils, and noncompetitively inhibits [125I]eotaxin binding and eotaxin-induced [35S]guanosine-5'-O-(3-thiotriphosphate) ([35S]GTPgammaS) binding. The proportion of [125I]eotaxin: [3H]Banyu (I) binding sites in eosinophils or transfectants was 35% or 13%, although both binding sites were overexpressed in transfectants. CCR3 spontaneously couples to G-proteins in CCR3 transfectants, demonstrated by changes in basal and eotaxin-induced [35S]GTPgammaS binding under reduced NaCl and GDP concentrations. Consequently, Banyu (I) was identified as an inverse agonist. In contrast, CCL18 and I-TAC (interferon-inducible T cell alpha-chemoattractant) were neutral antagonists, inhibiting eotaxin-induced [35S]GTPgammaS binding, with minimal effect on basal coupling of CCR3 to G proteins. Eotaxin, eotaxin-2 and monocyte chemoattractant protein (MCP)-4 are full agonists inducing [35S]GTPgammaS binding; eotaxin-3, MCP-3, RANTES (regulated on activation normal T cell expressed and secreted), vMIP-I (Kaposi's sarcoma-associated herpesvirus macrophage inflammatory protein-) and vMIP-II are partial agonists, indicating that this is a sensitive method to quantitate agonist efficacy.
PURPOSE: To describe our long-term experience with percutaneous access to continent urinary reservoirs for calculus removal. PATIENTS AND METHODS: A retrospective study of 13 procedures in 10 patients was performed. In 2 of the 13 procedures, access and calculus removal was performed in a single session. In the other 11 procedures, initial access was obtained using ultrasonography, fluoroscopy, and/or computed tomography. The patients then returned at a later date for a second step where the access was dilated and the calculi were removed. RESULTS: Access was achieved successfully in all cases with no complications. At mean follow-up time of 13.6 months (range 1-94 months) one patient had died of complications unrelated to her continent urinary reservoir. Another patient had been placed on suppressive antibiotics for recurrent calculi. The remaining patients were stone free and without late complication. CONCLUSIONS: Percutaneous removal of reservoir calculi can be performed safely, avoiding potential injury to the continence valve mechanism by a direct cystoscopic approach. We propose a two-stage procedure using CT guidance for initial access as the preferred technique.
Thirteen patients underwent placement of a balloon-expandable stent either at initial transjugular intrahepatic portosystemic shunt (TIPS) creation (n = 3) because of immediate technical failure of the Wallstent or at shunt revision because of failure of the Wallstent to reduce the portosystemic gradient <or=12 mm Hg despite overdilation (n = 2), stenosis of the Wallstent (n = 1), or parenchymal tract shunt stenosis (n = 7). The mean duration of primary patency when balloon-expandable stents were used at initial shunt creation was 162.7 days +/- 110.1 (SD). The time from initial TIPS creation or shunt revision with a Wallstent to placement of a balloon-expandable stent was 88.6 days +/- 115.3. The time from balloon-expandable stent placement to shunt revision (n = 6), most recent follow-up (n = 2), death (n = 1), or liver transplantation (n = 1) was 171.9 days +/- 121.6. No significant difference in shunt patency was found between the Wallstent and balloon-expandable stent (P =.31). Because no added benefit in patency was seen between the balloon-expandable stent and Wallstent, it appears that balloon-expandable stent placement is valuable only when placement of a Wallstent technically fails as a result of its limited radial force or when precision in placement is needed to treat a hepatic vein stenosis near the confluence with the inferior vena cava.
BACKGROUND: Our purpose was to compare the function and complications of two high-flow polyurethane hemodialysis catheters. METHODS: This prospective, randomized trial compared the Ash-Split (MedComp) and Opti-Flow (Bard Access Systems) catheters. All patients referred for tunneled hemodialysis catheter placement were offered entry in the study, provided they met inclusion criteria. Catheters were placed by interventional radiologists using ultrasound and fluoroscopic guidance. Procedure time and initial complications were recorded. Effective (QbEff) catheter flow rates and recirculation were studied at baseline, one month, three and six months using ultrasonic dilution (Transonic) at various pump speeds (Qb). Episodes of catheter malfunction and infection were recorded. Catheter removal or six months was the study endpoint. RESULTS: A total of 132 patients were enrolled in the trial. The groups did not differ as to age, sex distribution, height or weight (P> 0.05). Initial complications included kinking resulting in catheter failure (Optiflow N = 3), and tunnel bleeding (Optiflow N = 1; Ash N = 3). Adjusted mean flow rates (QbEff) at Qb300 were 299 mL/min Ash and 305 mL/min Optiflow (P = 0.06), at Qb400 were 365 mL/min Ash and 382 mL/min Optiflow (P = 0.01), and at QbMax were 414 mL/min Ash and 433 mL/min Optiflow (P = 0.03). Recirculation was significantly higher with the Optiflow catheter at most measurement points. Total late complications were lower in the Ash group (P = 0.04), and catheter survival was significantly higher in the Ash group (P = 0.02). CONCLUSIONS: Both catheters can deliver flow rates well beyond those recommended by the Dialysis Outcomes Quality Initiative. While the Optiflow delivered higher flow rates at some measurement points, this was offset by higher recirculation. The Ash catheter showed a long-term survival advantage and fewer late complications.