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Biomedical subjects

E Wang

Publications and source records attributed to E Wang.

459 records · Page 26Linked to original sources

Laser scanning cytometry evaluation of MART-1, gp100, and HLA-A2 expression in melanoma metastases.

Assessment of antigen expression by solid tumors has relied predominantly on immunohistochemistry, flow cytometry, and more recently quantitative real-time polymerase chain reaction. However, all these techniques present intrinsic limits. The laser scanning cytometer, by combining the properties of light and fluorescence microscopy with those of laser cytometry, can quantitatively and objectively analyze hypocellular samples such as fine-needle aspirates on an individual cell basis. To validate the fidelity of laser scanning cytometry for quantitative immunophenotyping of fine-needle aspirates, the authors measured the expression of the melanoma-associated antigens MART-1 and gp100 as well as HLA-A2, a HLA class 1 restriction element associated with their recognition by melanoma-specific T cells. Expression of melanoma antigens and HLA was measured by laser scanning cytometry and immunohistochemistry in fine-needle aspirates from melanoma metastases. In addition, transcription levels of both melanoma antigens were recorded by quantitative real-time polymerase chain reaction. A quantity of less than 1,000 cells per sample (average 682 cells) was sufficient for the analysis. Laser scanning cytometry estimates correlated with those of immunohistochemistry and quantitative real-time polymerase chain reaction for MART-1 and gp100. A good correlation in HLA-A2 detection by laser scanning cytometry and immunohistochemistry was also observed. Moreover, the laser scanning cytometer could discriminate subsets of cells from the same lesion with heterogeneous melanoma antigen expression, leading to the observation that cells with a DNA index greater than 2.5 expressed significantly less gp100. Thus, laser scanning cytometry yields detailed information on protein expression in individual cells and represents a new tool for dissecting the immune response in the tumor microenvironment.

Antigens, Neoplasm↗

Validation of changes in extracellular volume measured during hemodialysis using a segmental bioimpedance technique.

Sum of segmental bioimpedance analysis (BIAs) has recently been introduced as a more accurate estimation of extracellular volume (ECV). The current study was designed to compare the changes in ECV estimated by whole body bioimpedance (BIA(W)) and BIAs and to determine whether BIAs could be used to accurately estimate changes in ECV compared with the ultrafiltration volume (UFV) as measured by the dialysis machine. Ten men (age, 49 +/- 10 years; dry weight, 78 +/- 13 kg) were studied during hemodialysis (HD). A multifrequency bioimpedance analyzer (BIS4000B; Xitron Technologies, San Diego, CA) and a digital switch developed by the authors were used to automatically collect data from three body segments (arm, trunk, and leg) and the whole body using the stated technique. There was a significant difference in delta ECV(W) and UFV at the end of the treatment (2.75 +/- 0.74 L vs 3.95 +/- 0.73 L, p < 0.05) because fluid removal from the trunk has almost no effect on whole body resistance. However, delta ECVs was not significantly different from UFV (3.76 +/- 0.65 L vs 3.95 +/- 0.73 L, p = NS). BIAs is a more accurate approach to monitor changes in ECV during HD than is BIA(W) because changes in local resistance can be allocated to segments with uniform geometry and resistivity. This is a prerequisite for future studies on dynamics of regional fluid distribution during hemodialysis.

Biophysical Phenomena↗

Predictive value of access blood flow in detecting access thrombosis.

The aim of this study was to evaluate whether repeated measurement of access blood flow (Qac) using the ultrasound dilution technique could predict access failure in patients on hemodialysis. One hundred thirty-one patients were evaluated at intervals of 8 weeks for a period of 6 months. The incidence of thrombosis was determined within each study period. During the 6 month follow-up, 36 thrombotic events occurred in 27 of 68 polytetrafluoroethylene (PTFE) grafts, and only six thrombotic events in 5 of 63 arteriovenous (AV) fistulas. The relative risk for access thrombosis for patients with PTFE grafts was 5.6 times greater than for patients with AV fistulas. Qac was significantly lower in thrombotic compared with patent PTFE grafts (958 +/- 506 ml/min vs 1141 +/- 482 ml/min, p < 0.05). A significant relationship was found between the incidence of subsequent PTFE graft thrombotic events and Qac (p < 0.001). Compared with accesses with high blood flow (1100-1400 ml/min), the risk for subsequent thrombosis tripled in grafts with a Qac of less than 500 ml/min. This relationship was not seen with AV fistulas. In patent PTFE grafts, Qac remained unchanged within each 2 month interval, whereas it decreased in thrombotic PTFE grafts. Thus, repeated measurements of Qac have the potential to predict future access failure in PTFE grafts; however, an increased measuring frequency might improve the predictive value of graft failure with high Qac.

Female↗

Reducing atrazine losses: water quality implications of alternative runoff control practices.

Water quality is being affected by herbicides, some allegedly harmful to human health. Under scrutiny is atrazine (1-chloro-3-ethylamino-5-isopropylamino-2,4,6-triazine), a commonly used herbicide in corn (Zea mays L.) and sorghum [Sorghum bicolor (L.) Moench] production. Concentrations of soluble and adsorbed atrazine losses sometimes exceed the safe drinking water standard of 3 microg L(-1) established by the USEPA. This study assesses the protective implications of runoff control structures and alternative crop farming practices to minimize atrazine losses. Using a computerized simulation model, APEX, the following four practices were the most effective with respect to the average atrazine loss as a percent of the amount applied: (i) constructing sediment ponds, 0.09%; (ii) establishing grass filter strips, 0.14%; (iii) banding a 25% rate of atrazine, 0.40%; and (iv) constructing wetlands, 0.45%. Other atrazine runoff management options, including adoption of alternative tillage practices such as conservation and no-till as well as splitting applications between fall and spring, were marginally effective.

Agriculture↗

cDNA arrays and the enigma of melanoma immune responsiveness.

Melanoma is a cancer characterized by predisposition to natural and induced immune rejection. The occurrence of this phenomenon, however, remains sporadic and capricious while there is no clear understanding of the chain of events responsible. With the progression of the understanding of the molecular immunology of melanoma during the last decade, several theories suggesting immune tolerance (inadequate immune response) and/or immune escape (adaptation of cancer cells to a vigorous immune pressure) as possible modulators of melanoma growth have emerged. As the number of these hypotheses, mostly based on molecular or pre-clinical models, has increased exponentially, the relevance of each individual one has been conversely decreasing leaving the observer without confidence that a theory likely to explain this phenomenon can be soon formulated. In this review, we will discuss a direct approach whereby new theories might be identified by direct observation of human phenomena as they occur by depiction of molecular portraits of melanoma lesions using global transcript analysis.

Gene Expression Profiling↗

Cytokines and immune response in the tumor microenvironment.

Over the last few decades a wealth of evidence has been gathered on the potential role that the immune system (IS) can play in the fight against cancer. Together with cell surface adhesion molecules, cytokines (CKs) mediate the activities of IS cells. Therefore, CK kinetics may represent a mirror of the immunologic phenomena occurring in the tumor microenvironment, where immune and malignant cells interact. Yet, CKs are currently used in a clinical setting to polarize the immune response against cancer. Despite the large amount of information available on IS physiology, little is known about the role of CKs in modulating the effectiveness of immunotherapy clinical trials aimed at the treatment of patients with cancer. This underscores our relative ignorance about the complex cascade of events that lead to tumor rejection. Here, we review the properties of some CKs believed to be particularly relevant to tumor immunology (i.e., interleukin [IL]-10, transforming growth factor-beta, interferon-gamma, IL-2, IL-4, and IL-12). We summarized the experience gained with these CKs in vitro, in animal models, and in human beings to illustrate the achievements, the controversies, and the challenges that characterize this fascinating field of oncology. In addition, we added a short section in which a broad view of CKs released in the tumor microenvironment is proposed to underline the variety of factors that contribute to the complexity of tumor-IS interactions.

Cytokines↗

Dynamic internal compliance of a vascular prosthesis.

A new technique was used to measure dynamic internal compliance of a blood vessel or vascular graft subjected to dynamic internal pressure. The internal compliance can be broken into three categories: the volumetric compliance (CV), defined as (dV/V)/dP; the longitudinal compliance (CL), defined as (dL/L)/dP; and radial compliance (CR), defined as (dR/R)/dP. It can be shown mathematically that CV = 2 CR + CL. Thus, measuring any two of the three entities will also give the value for the third. A Dynatek DCT1 dynamic compliance tester was used for measuring the compliance of DenaflexTM biologic grafts and fresh bovine internal thoracic arteries, from which the Denaflex grafts were obtained by fixation. Volumetric compliance was obtained with the test sample mounted in a loose loop that allowed the sample to move both radially and longitudinally. By mounting the sample in a straight fashion that limited longitudinal movement, the radial compliance was determined. The longitudinal compliance was then calculated from the above relationship. Test results show that the fresh bovine artery had an average volumetric compliance of 26.1%/100 mmHg, radial compliance of 9.5%/100 mmHg, and longitudinal compliance of 7.2%/100 mmHg. The Denaflex vascular graft showed a reduction in longitudinal and radial compliance, compared to the fresh raw artery, as a result of extensive fixation.

Animals↗

Effects of flavonoids on cytochrome P450-dependent acetaminophen metabolism in rats and human liver microsomes.

Flavonoids are widely occurring in our diet. In this study, the effects of nine flavonoids on acetaminophen (APAP) oxidation and related cytochrome P450 (P450) enzyme activities were investigated. With rat liver microsomes, APAP oxidation was stimulated 2- to 4-fold by 50 microM flavone or tangeretin and inhibited 65% by myricetin or quercetin. Only a slight inhibition of APAP oxidation was caused by naringenin. All the effects cited herein were from experiments with 50 microM flavonoids. With human liver microsomal samples that had high P450 3A4 activity, APAP oxidation was stimulated 1.6- to 3.0-fold by tangeretin, nobiletin, and flavone, but inhibited 40-60% by myricetin and quercetin. With human P450 1A2 expressed in Hep G2 cells, APAP oxidation was inhibited completely by flavone or quercetin. With expressed P450 3A4, this reaction was stimulated 4-fold by flavone, but inhibited 60% by quercetin. The expressed human P450 2E1-dependent APAP oxidation was only slightly affected by flavone and quercetin. The mechanisms of the inhibition and stimulation were complex and varied with the different P450 forms and flavonoids used in the system. The O-deethylation of ethoxyresorufin in rat liver microsomes was effectively inhibited by myricetin (IC50, 15 microM) and moderately inhibited by flavone, tangeretin, and quercetin (IC50, 50-80 microM); with P450 1A2 in Hep G2 cell microsomes, the activity was markedly inhibited by flavone (IC50, 2.5 microM). The microsomal P450s 2E1 and 3A activities were inhibited by myricetin (IC50, 85-90 microM), but not effectively inhibited by other flavonoids.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetaminophen↗

Expression of an antisense transforming growth factor-beta1 transgene reduces tumorigenicity of EMT6 mammary tumor cells.

Transforming growth factor-beta (TGF-beta) is a potent immunosuppressive cytokine produced by many tumor cells. Secretion of TGF-beta by malignant cells may therefore be a mechanism by which tumor cells escape destruction by tumor-specific T lymphocytes. In order to evaluate the role of tumor-derived TGF-beta on tumor progression, we have inhibited the production of this cytokine by introducing a gene encoding antisense TGF-beta1 into the EMT6 murine mammary tumor cell line using a retroviral vector (Las-TGF-beta1SN). EMT6 cells transduced with this vector (EMT6as-TGF-beta1) stably expressed the antisense gene and secreted 52% less TGF-beta than did tumor cells transduced with the backbone vector alone. Supernatant fluid recovered from tumor cells expressing the antisense TGF-beta1 gene also exhibited a decreased capacity to inhibit alloantigen-specific cytotoxic T-cell responses in vitro. Furthermore, tumor growth in mice injected with EMT6as-TGF-beta1 tumor cells was inhibited compared to mice injected with control tumor cells. These results demonstrate that expression of antisense TGF-beta1 by transduced EMT6 cells decreases their tumorigenicity and suggest that this approach of eliminating immune suppression is a potentially useful strategy to enhance antitumor responses.

Animals↗