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At least 1,297 records · Page 72Linked to original sources

PubMed1992

Monoclonal antibodies to the spike protein of feline infectious peritonitis virus mediate antibody-dependent enhancement of infection of feline macrophages.

Antibody-dependent enhancement of virus infection is a process whereby virus-antibody complexes initiate infection of cells via Fc receptor-mediated endocytosis. We sought to investigate antibody-dependent enhancement of feline infectious peritonitis virus infection of primary feline peritoneal macrophages in vitro. Enhancement of infection was assessed, after indirect immunofluorescent-antibody labelling of infected cells, by determining the ratio between the number of cells infected in the presence and absence of virus-specific antibody. Infection enhancement was initially demonstrated by using heat-inactivated, virus-specific feline antiserum. Functional compatibility between murine immunoglobulin molecules and feline Fc receptors was demonstrated by using murine anti-sheep erythrocyte serum and an antibody-coated sheep erythrocyte phagocytosis assay. Thirty-seven murine monoclonal antibodies specific for the nucleocapsid, membrane, or spike proteins of feline infectious peritonitis virus or transmissible gastroenteritis virus were assayed for their ability to enhance the infectivity of feline infectious peritonitis virus. Infection enhancement was mediated by a subset of spike protein-specific monoclonal antibodies. A distinct correlation was seen between the ability of a monoclonal antibody to cause virus neutralization in a routine cell culture neutralization assay and its ability to mediate infection enhancement of macrophages. Infection enhancement was shown to be Fc receptor mediated by blockade of antibody-Fc receptor interaction using staphylococcal protein A. Our results are consistent with the hypothesis that antibody-dependent enhancement of feline infectious peritonitis virus infectivity is mediated by antibody directed against specific sites on the spike protein.

Animals↗

PubMed2004

Dynamic imaging of pancreatic diseases by contrast enhanced coded phase inversion harmonic ultrasonography.

BACKGROUND: Coded phase inversion harmonic ultrasonography, a newly available sonographic technique, enables visualisation of slow flow in minute vessels in a real time fashion with the use of a sonographic contrast agent containing monosaccharide. Our purpose was to employ this novel technique to observe microvessels in pancreatic tumours. SUBJECTS AND METHODS: Sixty five patients with suspicious pancreatic tumours received contrast enhanced coded phase inversion harmonic ultrasonography, contrast enhanced computed tomography, and endosonography. Final diagnoses based on histological findings were pancreatic ductal carcinomas in 49 patients, inflammatory pseudotumours with chronic pancreatitis in seven, and endocrine tumours in nine. For contrast enhanced coded harmonic ultrasonography, Levovist, a contrast agent, was injected intravenously as a bolus. When the first microbubble signal appeared in the pancreas, images of the ideal scanning plane were displayed in a real time continuous fashion (vessel images). Subsequently, interval delay scanning (perfusion images) was taken to demonstrate parenchymal flow. Tumour vascularity was evaluated by using the two types of imaging. Sensitivities for depicting pancreatic tumours were compared between three examinations. RESULTS: Contrast enhanced ultrasonography demonstrated tumour vessels in 67% of pancreatic ductal carcinomas, although most were relatively hypovascular compared with the surrounding pancreatic tissue. The vascular patterns of tumours obtained by contrast enhanced ultrasonography were closely correlated with those obtained by contrast enhanced computed tomography. Values for sensitivity in depicting pancreatic tumours of 2 cm or less in size were 68% for contrast enhanced computed tomography, 95% for endosonography, and 95% for contrast enhanced ultrasonography. CONCLUSION: Contrast enhanced coded phase inversion harmonic ultrasonography successfully visualised fine vessels in pancreatic tumours and may play a pivotal role in the depiction and differential diagnosis of pancreatic tumours.

Adult↗

PubMed2002

Radio-frequency ablation of VX2 rabbit tumors: assessment of completeness of treatment by using contrast-enhanced harmonic power Doppler US.

PURPOSE: To assess contrast material-enhanced harmonic power Doppler and fundamental color Doppler ultrasonography (US) in the detection of residual viable tumor tissue after radio-frequency (RF) ablation in tumors embedded in fat. MATERIALS AND METHODS: Twenty-eight VX2 tumors were implanted into the retroperitoneum of 14 rabbits. Tumors were examined with contrast-enhanced fundamental color Doppler US and harmonic power Doppler US before and 10 minutes after RF ablation. Saline-enhanced RF ablation (30 mL/h) was performed over 10 minutes with 28-W RF power. Follow-up included repeat US examinations. Necropsies and histopathologic assessment were performed after detection of residual untreated tumor at US or 3 weeks after ablation. RESULTS: VX2 tumors reached a mean size of 21 mm +/- 9 (SD) (size range, 6-43 mm) 25 days after implantation. All tumors larger than 31 mm showed signs of central necrosis at US. Before ablation, intense vascularity was detected in all tumors with both contrast-enhanced US modes. Histopathologic assessment at the end of the follow-up period revealed local relapses due to incomplete ablation in 14 (50%) of 28 cases. Detection of residual tumor was missed in all cases with contrast-enhanced color Doppler US. Contrast-enhanced harmonic power Doppler US depicted residual flow in 12 of the 14 cases (sensitivity, 86%) in which local relapses occurred. There was a significant (P <.005, McNemar test) improvement in detection of residual tumor when the harmonic power Doppler mode was used. CONCLUSION: Contrast-enhanced harmonic power Doppler US has greater sensitivity than contrast-enhanced color Doppler US for detecting residual VX2 tumor following ablation. Therefore, contrast-enhanced harmonic power Doppler US may be a useful additional method for the detection of residual tumors after RF ablation.

Animals↗

PubMed2003

Carotid artery stenosis: accuracy of contrast-enhanced MR angiography for diagnosis.

PURPOSE: To assess accuracy of contrast material-enhanced magnetic resonance (MR) angiography as compared with three-dimensional (3D) time-of-flight (TOF) MR angiography and reference digital subtraction angiography (DSA) in diagnosis of carotid artery stenosis. MATERIALS AND METHODS: Enhanced and 3D TOF MR angiography and DSA were performed in 51 consecutive patients suspected of having carotid artery stenosis at duplex ultrasonography. Stenoses were measured by two independent observers blinded to clinical information and other test results. Pearson correlation coefficients were used, and kappa for interobserver variabilities was estimated. Sensitivity and specificity of enhanced and 3D TOF MR angiography were calculated and compared with those of DSA. RESULTS: Pearson correlation coefficients were 0.94 (P <.01) for enhanced angiography versus DSA, 0.92 (P <.01) for 3D TOF angiography versus DSA, and 0.93 (P <.01) for enhanced versus 3D TOF angiography for observer 1 and 0.94 (P <.01), 0.95 (P <.01), and 0.94 (P <.01), respectively, for observer 2. kappa statistics were 0.81 for enhanced angiography, 0.79 for 3D TOF angiography, and 0.78 for DSA. Stenosis measurements of observer 1 at enhanced MR angiography, with inclusion of carotid arteries on the symptomatic side only, compared with those of DSA yielded a sensitivity of 90% (95% CI: 68%, 99%) and a specificity of 77% (95% CI: 55%, 92%). 3D TOF angiography yielded a sensitivity of 86% (95% CI: 67%, 97%) and a specificity of 73% (95% CI: 50%, 89%) compared with those of DSA. For observer 2, sensitivity and specificity for enhanced angiography were 91% (95% CI: 70%, 99%) and 76% (95% CI: 52%, 91%), respectively, and 90% (95% CI: 68%, 99%) and 77% (95% CI: 51%, 92%), respectively, for 3D TOF angiography. CONCLUSION: Accuracy of enhanced MR angiography in diagnosis of severe stenosis is similar to that of 3D TOF MR angiography.

Adult↗

PubMed2003

Discrimination of prostate cancer from normal peripheral zone and central gland tissue by using dynamic contrast-enhanced MR imaging.

PURPOSE: To evaluate which parameters of dynamic magnetic resonance (MR) imaging and T2 relaxation rate would result in optimal discrimination of prostatic carcinoma from normal peripheral zone (PZ) and central gland (CG) tissues and to correlate these parameters with tumor stage, Gleason score, patient age, and tumor markers. MATERIALS AND METHODS: Of 58 patients with prostatic carcinoma, 36 were included for analysis. Patients underwent MR imaging at 1.5 T with an endorectal-pelvic phased-array coil and subsequently underwent prostatectomy. A T2-weighted turbo spin-echo sequence, an intermediate-weighted sequence, and a fast T1-weighted gradient-echo sequence (seven sections in 2.03 seconds) during bolus injection of 0.1 mmol gadopentetate dimeglumine per kilogram of body weight were performed. Contrast agent concentration-time curves were obtained for prostatic carcinoma and normal PZ and CG tissue by using whole-mount sections to guide placement of regions of interest. Onset time, time to peak, peak enhancement, relative peak enhancement, washout, and T2 relaxation rates were calculated. Multivariate receiver operating characteristic analysis was performed with and without relative peak enhancement. RESULTS: Results of multivariate receiver operating characteristic analysis showed that relative peak enhancement demonstrated the highest area under the receiver operating characteristic curve (AUC) in the PZ and the CG (AUC = 0.93, 0.82). Results of multivariate analysis without relative peak enhancement showed that relative peak enhancement in the PZ and washout in the CG demonstrated the highest AUC (AUC = 0.9, 0.81). Pearson correlation coefficients between the dynamic parameters or T2 relaxation rates in carcinoma and the tumor stage, Gleason score, patient age, and tumor markers ranged between 0.02 and 0.44. CONCLUSION: The optimal parameter for discrimination of prostatic carcinoma in the PZ and CG was relative peak enhancement. If relative peak enhancement was not used, then peak enhancement was optimal in the PZ, and washout was optimal in the CG. Poor-to-moderate correlation was present between the dynamic parameters or T2 relaxation rate in carcinoma and the tumor stage, Gleason score, patient age, tumor volume, and prostate-specific antigen.

Aged↗

PubMed2003

Bone marrow with diffuse tumor infiltration in patients with lymphoproliferative diseases: dynamic gadolinium-enhanced MR imaging.

PURPOSE: To evaluate gadolinium enhancement of bone marrow in patients with lymphoproliferative diseases and diffuse bone marrow involvement. MATERIALS AND METHODS: Dynamic contrast material-enhanced magnetic resonance (MR) imaging of the thoracolumbar spine was performed in 42 patients with histologically proved diffuse bone marrow involvement and newly diagnosed myeloma (n = 31), non-Hodgkin lymphoma (n = 8), or Hodgkin disease (n = 3). The maximum percentage of enhancement (Emax), enhancement slope, and enhancement washout were determined from enhancement time curves (ETCs). A three-grade system for scoring bone marrow involvement was based on the percentage of neoplastic cells in bone marrow samples. Quantitative ETC values for the 42 patients were compared with ETC values for healthy subjects and with grades of bone marrow involvement by using mean t test comparisons. Receiver operating characteristic (ROC) analysis was conducted by comparing Emax values between patients with and those without bone marrow involvement. Baseline and follow-up MR imaging findings were compared in nine patients. RESULTS: Significant differences in Emax (P <.001), slope (P <.001), and washout (P =.005) were found between subjects with normal bone marrow and patients with diffuse bone marrow involvement. ROC analysis results showed Emax values to have a diagnostic accuracy of 99%. Emax, slope, and washout values increased with increasing bone marrow involvement grade. The mean Emax increased from 339% to 737%. Contrast enhancement decreased after treatment in all six patients who responded to treatment but not in two of three patients who did not respond to treatment. CONCLUSION: Dynamic contrast-enhanced MR images can demonstrate increased bone marrow enhancement in patients with lymphoproliferative diseases and marrow involvement.

Adult↗

PubMed1993

Dynamic contrast-enhanced CT of the liver: comparison of contrast medium injection rates and uniphasic and biphasic injection protocols.

To determine the effect of contrast medium injection rate and biphasic versus uniphasic bolus on hepatic parenchymal enhancement at computed tomography (CT), 105 patients were randomized into four protocols. Hepatic and aortic enhancement curves were measured for each protocol. The optimal scanning interval (the duration between onset of a desired hepatic enhancement threshold and decline of enhancement below the threshold or onset of the equilibrium phase) and contrast enhancement index (CEI) (area under the hepatic enhancement curve above a desired threshold during the nonequilibrium phase) were the primary determinants of merit. Measurements were performed at hepatic enhancement thresholds between 10 and 60 HU. At most thresholds of hepatic enhancement, optimal scanning intervals were significantly longer and CEIs were significantly higher for the biphasic protocols than for the uniphasic protocols (P < .03). A biphasic injection with a high initial flow rate produces high peak contrast enhancement but delayed onset of equilibrium and thus provides a longer optimal scanning interval.

Adult↗

PubMed1995

Enhancement of the intracranial arterial wall at MR imaging: relationship to cerebral atherosclerosis.

PURPOSE: To investigate the relationship between intracranial arterial wall enhancement and atherosclerosis. MATERIALS AND METHODS: Intracranial vertebral arteries of 30 patients and carotid arteries of 62 patients were studied with spin-echo magnetic resonance imaging with contrast enhancement and spatial presaturation. Arterial wall enhancement was graded as follows: stage 1, no substantial enhancement; stage 2, faint or thin area of enhancement; stage 3, definite and thick area of enhancement. RESULTS: In vertebral arteries, stage 3 enhancement was seen in 11 patients (mean age, 73.7 years) and stage 1 in eight (mean age, 56.4 years). In carotid arteries, stage 3 enhancement was seen in 13 patients (mean age, 71.0 years) and stage 1 in 21 patients (mean age, 39.0 years). In both arteries, stage was well correlated with age (P < .05). CONCLUSION: Arterial wall enhancement is related to aging and is probably due to neovascularity in association with atherosclerotic plaques. This finding may permit assessment of intracranial atherosclerosis and other vascular diseases.

Adult↗

PubMed1995

Adrenal gland enhancement at MR imaging with Mn-DPDP.

PURPOSE: To determine adrenal gland enhancement with manganese (II) N,N' dipyridoxylethylenediamine-N,N'-diacetate 5,5'bis(phosphate) (DPDP) at magnetic resonance imaging. MATERIALS AND METHODS: After phase III trials, fat-suppressed, motion-compensated, T1-weighted spin-echo images (repetition time, 500 msec; echo time, 12 msec) of 13 consecutive patients were obtained at 1.5 T at one site, prior to and approximately 30 minutes after intravenous administration of 5 mumol/kg Mn-DPDP. Images were analyzed visually and by means of region-of-interest measurements, normalized to spleen. With data added from three more sites, enhancement of three adrenal adenomas and two metastases was analyzed. RESULTS: Twenty-five of 26 adrenal glands were depicted on MR images, and all showed enhancement. Mean adrenal enhancement (38%) was comparable to mean enhancement of liver (46%), pancreas (25%), and renal cortex (58%). Adrenal cortex and medulla could not be distinguished in nonenhanced or enhanced images. All three adrenal adenomas enhanced by more than 40%, but the metastases did not enhance. CONCLUSION: Functioning adrenal tissue (glands and at least some adenomas) are enhanced with Mn-DPDP.

Adenoma↗

PubMed1998

Aortic and hepatic contrast medium enhancement at CT. Part II. Effect of reduced cardiac output in a porcine model.

PURPOSE: To investigate how reduction in cardiac output affects the magnitude and timing of aortic and hepatic contrast medium enhancement during abdominal computed tomography (CT). MATERIALS AND METHODS: Eight 20-30-kg pigs underwent CT before and after pharmacologic reduction of cardiac output (measured by means of thermodilution). Each CT study consisted of 53 dynamic images acquired every 5 seconds at a fixed level through the midliver after intravenous injection of contrast medium (concentration, 282 mg of iodine per milliliter; dose, 2 mL per kilogram of body weight; injection rate, 2 mL/sec). Curves of contrast medium enhancement versus time were measured. Changes in the magnitude and timing of aortic and hepatic enhancement were compared with the reduction in cardiac output. RESULTS: With reduction in cardiac output, the time from the injection start to the arrival of the contrast medium bolus in the aorta (P < .01) and the times from injection completion to peak aortic (P < .01) and peak hepatic (P < .01) enhancement increased. As cardiac output decreased, peak aortic enhancement increased proportionally (P < .01). Peak hepatic enhancement increased only slightly and correlated weakly with the decrease in cardiac output (P = .07). CONCLUSION: As cardiac output decreases, the times to the arrival of the contrast medium bolus in the aorta and to peak aortic and hepatic enhancement increase. Reduction in cardiac output results in a substantial increase in peak aortic enhancement but not in peak hepatic enhancement.

Animals↗

PubMed1999

Single-dose breath-hold gadolinium-enhanced three-dimensional MR angiography of the renal arteries.

PURPOSE: To evaluate the quality of single-dose breath-hold three-dimensional (3D) magnetic resonance (MR) angiography of the renal arteries optimized with a 1-mL test bolus timing examination. MATERIALS AND METHODS: Three-dimensional spoiled gradient-echo imaging (3.8-4.2/1.3-1.7 [repetition time msec/echo time msec], 25 degrees-40 degrees flip angle) was performed in 60 patients after administration of gadopentetate dimeglumine (average dose, 0.11 mmol/kg). Synchronization of contrast material administration with data acquisition was achieved with a 1-mL test dose of contrast material to estimate patient circulation parameters. Image quality was assessed by using contrast-to-noise (CNR), relative vascular enhancement, and venous-to-arterial enhancement ratios and subjective scoring of arterial and venous enhancement. The effect of the contrast material injection rate and the influence of breath holding during the timing examination also were examined. RESULTS: Overall, of 60 studies, 58 were diagnostic and 56 demonstrated excellent arterial enhancement. Venous enhancement was seen in eight studies. The average aortic relative vascular enhancement (+/- SD) was 14.6 +/- 5.9, with an aorta-to-inferior vena cava (IVC) CNR of 69.7 +/- 43.9. The IVC-to-aorta venous-to-arterial enhancement ratio averaged 0.08 +/- 0.16. There was no significant difference in image quality based on injection rates or the performance of breath holding during the timing examination (P > .1). CONCLUSION: Breath-hold gadolinium-enhanced renal MR angiography free of venous enhancement can be performed consistently and reliably with 20 mL of contrast material when studies are synchronized to patient circulation time by using a timing examination.

Contrast Media↗

PubMedDate not supplied

Blood flow patterns in focal liver lesions at microbubble-enhanced US.

Noninvasive diagnosis of liver lesions is usually performed with contrast material-enhanced computed tomography (CT) and magnetic resonance (MR) imaging and is based on enhancement features of the arterial and portal venous phases. Ultrasonography (US) is often limited in characterizing liver lesions because color and spectral Doppler US provide limited vascular information in large patients and in small or deep lesions. However, microbubble contrast agents, together with specialized US techniques, now allow diagnosis of liver lesions based on morphologic evaluation of lesion vascularity and visualization of specific enhancement features. Microbubble contrast agents are purely intravascular, easy to administer, and well tolerated and allow sensitive real-time evaluation of blood flow in hepatic lesions. During the portal venous phase, benign lesions (eg, hemangioma, focal nodular hyperplasia) typically enhance more than the liver, whereas malignant lesions (eg, hepatocellular carcinoma, metastases) enhance less. Microbubble-enhanced US allows characterization of very small lesions that may not be accurately characterized with CT or MR imaging. Findings from initial studies suggest that microbubble-enhanced US of the liver provides enhancement information comparable to that provided by contrast-enhanced CT and MR imaging, along with real-time morphologic evaluation of lesion vascularity.

Acoustics↗

PubMedDate not supplied

Normal magnetic resonance contrast enhancement of extraocular muscles: a quantitative analysis.

The purpose was to evaluate quantitatively the magnetic resonance contrast enhancement of normal extraocular muscles and the use of temporal muscles as a reference of enhancement. Eighty extraocular and 20 temporal muscles were taken into analysis. Before contrast administration, mean intensity of extraocular muscles was found to be higher than that of temporal muscles (p < 0.000). With contrast agent, all extraocular muscles were enhanced more (111% enhancement) than the temporal muscles (45% enhancement, p < 0.000). Lateral recti had the lowest signal intensity, both in pre- and postcontrast images (p < 0.005). Normal extraocular muscles showed prominent enhancement on contrast-enhanced T(1)-weighted images. Temporal muscles were also enhanced in all subjects, urging the observers to compare the enhancement of extraocular muscles not with the latter.

Adolescent↗

PubMed2006

Outcomes of enhanced counseling services provided to adults through Project Liberty.

OBJECTIVE: Project Liberty provided brief crisis counseling to 753,015 residents of New York City and surrounding counties after the attacks on the World Trade Center. Most regained predisaster functioning after counseling. For those who did not, Project Liberty provided enhanced services by specially trained, licensed mental health professionals. Individuals receiving crisis counseling and enhanced services responded to confidential telephone interviews about 18 and 24 months, respectively, after the attacks. Impairment was compared between groups to determine whether enhanced services recipients reported improved functioning and fewer symptoms of depression, posttraumatic stress, and complicated grief. METHODS: Crisis counseling recipients (N=153) were interviewed once and enhanced services recipients (N=76) were interviewed twice about symptomatology and daily functioning. RESULTS: The samples did not differ in age or gender. Significantly greater proportions of enhanced services recipients reported knowing someone who died as a result of the attacks, having been involved in rescue efforts, or having lost their job because of the attacks. Compared with crisis counseling respondents, enhanced services recipients at their first interview reported significantly more symptoms of depression, grief, and traumatic stress and significantly poorer daily functioning in five life areas. At follow-up, enhanced services respondents reported significant improvement in three of five functioning domains, significantly fewer symptoms of depression and grief, and marginally less traumatic stress. CONCLUSIONS: Recipients of enhanced services were more impaired than people who received only crisis counseling. On the basis of reports from service recipients, meaningful improvements in functioning and symptoms may be associated with the receipt of enhanced services.

Activities of Daily Living↗

PubMed2006

The saponin-mediated enhanced uptake of targeted saporin-based drugs is strongly dependent on the saponin structure.

Saponins are a group of plant glycosides consisting of a steroid or triterpenoid aglycone to which one or more sugar chains are attached. They exhibit cell membrane-permeabilizing properties and, thus, have been investigated for their therapeutic potential. Recently, at a non-permeabilizing concentration saponinum album from Gypsophila paniculata L. has been described to enhance the cytotoxicity of a chimeric toxin in a cell culture model. To elucidate whether this enhancing effect is also mediated by other saponins, we analyzed the ability of seven different saponins to enhance the cytotoxicity of a targeted chimeric toxin. The chimeric toxin is composed of saporin, a plant ribosome-inactivating toxin, a cleavable adapter, and human epidermal growth factor (EGF). Cytotoxicity on EGF receptor (EGFR)-bearing cells was analyzed both alone and after combined application of saponin and chimeric toxin. Only two of the tested saponins, quillajasaponin and saponinum album, enhanced cytotoxicity by more than 1,000-fold, whereas the enhancement factors of the other saponins were only approximately 10-fold. In contrast to saponinum album, quillajasaponin enhanced the cytotoxicity both on control cells lacking EGFR and on target cells, indicating that, in this case, the enhancement is not target cell receptor specific. This is also the case for some of the saponins with low enhancement factors. Saponinum album resulted in a more than 13,600-fold receptor-specific enhancement, decreasing the 50% inhibitory concentration (IC(50)) from 2.4 nM to 0.18 pM, which renders it the best option to promote saporin-3-based drug uptake while retaining specificity for the EGFR.

Animals↗

PubMed1988

Mapping gold-labeled IgE receptors on mast cells by scanning electron microscopy: receptor distributions revealed by silver enhancement, backscattered electron imaging, and digital image analysis.

Immunogold labeling and silver enhancement techniques are widely used to determine density and distribution of cell membrane receptors by light and transmission electron microscopy. However, these techniques have not been widely used for receptor detection by scanning electron microscopy. We used antigen- or protein A-conjugated colloidal gold particles, together with silver enhancement, sequential secondary and back-scattered electron imaging (SEI and BEI), and digital image processing, to explore cell surface distribution of IgE-receptor complexes on RBL-2H3 cells, a rat leukemia line that provides a model for the study of mucosal mast cells. Cells were first incubated with a monoclonal antidinitrophenol IgE (anti-DNP-IgE) that binds with high affinity to cell surface IgE receptors. The resulting IgE-receptor complexes were cross-linked either with the multivalent antigen, DNP-BSA-gold, or with a polyclonal anti-IgE antibody. Antibody-treated cells were labeled after fixation with protein A-gold. Fixed, gold-labeled cell monolayers were silver enhanced (or not), dehydrated, critical point-dried, and coated with gold-palladium (for SEI analysis) or carbon (for combined SEI/BEI analysis). They were observed in an Hitachi S800 SEM equipped with a field emission tip and a Robinson backscattered electron detector. An image processor (MegaVision 1024XM) digitized images directly from the S800 microscope at 500-1000 line resolution. Silver enhancement significantly improves detection of gold particles in both SEI and BEI modes of SEM. On gold-palladium-coated samples, 20-nm particles are resolved by SEI after enhancement. BEI resolves 15-nm particles without enhancement and 5- or 10-nm particles are resolved by BEI on silver-enhanced, carbon-coated samples. Neither BEI nor SEI alone can yield high resolution topographical maps of receptor distribution (BEI forms images on the basis of atomic number contrast which reveals gold but not surface features). Image analysis techniques were therefore introduced to digitize, enhance, and process BEI and SEI images of the same field of view. The resulting high-contrast, high-resolution images were superimposed, yielding well-resolved maps of the distribution of antigen-IgE-receptor complexes on the surface of RBL-2H3 mast cells. The maps are stored in digital form, as required for computer-based quantitative morphometric analyses. These techniques of silver enhancement, combined BEI/SEI imaging, and digital image analysis can be applied to analyze density and distribution of any gold-labeled ligand on its target cell.

Cell Line↗

PubMed2001

Multiple redundant sequence elements within the fission yeast ura4 replication origin enhancer.

BACKGROUND: Some origins in eukaryotic chromosomes fire more frequently than others. In the fission yeast, Schizosaccharomyces pombe, the relative firing frequencies of the three origins clustered 4-8 kbp upstream of the ura4 gene are controlled by a replication enhancer - an element that stimulates nearby origins in a relatively position-and orientation-independent fashion. The important sequence motifs within this enhancer were not previously localized. RESULTS: Systematic deletion of consecutive segments of approximately 50, approximately 100 or approximately 150 bp within the enhancer and its adjacent core origin (ars3002) revealed that several of the approximately 50-bp stretches within the enhancer contribute to its function in partially redundant fashion. Other stretches within the enhancer are inhibitory. Some of the stretches within the enhancer proved to be redundant with sequences within core ars3002. Consequently the collection of sequences important for core origin function was found to depend on whether the core origin is assayed in the presence or absence of the enhancer. Some of the important sequences in the core origin and enhancer co-localize with short runs of adenines or thymines, which may serve as binding sites for the fission yeast Origin Recognition Complex (ORC). Others co-localize with matches to consensus sequences commonly found in fission yeast replication origins. CONCLUSIONS: The enhancer within the ura4 origin cluster in fission yeast contains multiple sequence motifs. Many of these stimulate origin function in partially redundant fashion. Some of them resemble motifs also found in core origins. The next step is to identify the proteins that bind to these stimulatory sequences.

Journal Article↗

PubMed1996

Vasoactive intestinal peptide enhances immunoglobulin production and growth in human plasma cells via mechanisms that may involve protein kinase C.

The effects of various neuropeptides on human plasma cells were studied. Of the various neuropeptides tested, vasoactive intestinal peptide (VIP) enhanced Ig production and growth in human plasma cell lines, IM-9 and AF-10, and in plasma cells generated in vivo (four out of four patients with plasma cell leukemia) and in vitro. In contrast, other neuropeptides (neuropeptide Y, somatostatin, substance P, peptide YY, neurokinin A, calcitonin gene-related peptide, chole-cystokinin octapeptide, and beta-endorphin) were ineffective. Moreover, VIP-induced enhancement was specifically blocked by VIP receptor antagonist. Among the various cytokines, IL-6, GH, and insulin-like growth factor I (IGF-I) also enhanced Ig production and thymidine uptake in plasma cells. However, VIP-induced enhancement was not mediated by IL-6, GH, or IGF-I because antibodies to these cytokines failed to block VIP-induced enhancement. Phorbol 12,13 dibutyrate enhanced Ig production and thymidine uptake in plasma cells, and the Phorbol 12,13 dibutyrate-induced enhancement was blocked by H7 (a protein kinase C inhibitor) but not by H8 (a protein kinase A inhibitor). Similarly, VIP-induced enhancement was blocked by H7 but not by H8. Collectively, VIP enhances plasma cell responses via mechanisms that may involve protein kinase C.

Cell Division↗