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1.
Dermatology ; 217(2): 164-8, 2008.
Article in English | MEDLINE | ID: mdl-18525203

ABSTRACT

BACKGROUND/OBJECTIVE: Acne prevalence studies often use subject self-report as data source. Our aim was to evaluate the validity of acne self-report. METHODS: Responses of university students to an acne questionnaire were compared to the trained observer's concurrent examination of acne. The validity of self-report was measured by sensitivity, specificity, positive predictive value and negative predictive value. Agreement was measured by Cohen's kappa and correct classification percentage. RESULTS: The sensitivity of self-report was 0.55 (95% CI 0.47-0.63), the specificity was 0.72 (95% CI 0.63-0.80), the positive predictive value was 0.70 (95% CI 0.61-0.78), and the negative predictive value was 0.57 (95% CI 0.49-0.65). Cohen's kappa was 0.26 (95% CI 0.15-0.38) and correct classification percentage was 63. CONCLUSIONS: Validity of self-report was moderate at best and agreement was fair, indicating that college students could not accurately report that they have acne. This is likely not sufficient for clinical or research activities or to assure that individuals who self-guide their acne therapy actually have acne.


Subject(s)
Acne Vulgaris/diagnosis , Acne Vulgaris/epidemiology , Health Knowledge, Attitudes, Practice , Self Concept , Acne Vulgaris/drug therapy , Adolescent , Adult , Age Factors , Anti-Bacterial Agents/therapeutic use , Confidence Intervals , Cross-Sectional Studies , Dermatologic Agents/therapeutic use , Female , Humans , Male , Prevalence , Probability , Sensitivity and Specificity , Severity of Illness Index , Sex Factors , Surveys and Questionnaires , Treatment Outcome
2.
Int J Cancer ; 123(2): 464-475, 2008 Jul 15.
Article in English | MEDLINE | ID: mdl-18449880

ABSTRACT

This study investigates the role of tumor nitric oxide (NO) and vascular regulation in tumor ulceration following high-dose tumor necrosis factor-alpha (TNF) treatment. Using TNF-responsive (MethA) and nonresponsive (LL2) mouse tumors, tumor NO concentration was measured with an electrochemical sensor and tumor blood flow by Doppler ultrasound. Mice were also pretreated with a selective inducible nitric oxide synthase (iNOS) inhibitor, 1400 W. Tumors harvested from TNF-treated mice were cryosectioned and immunostained for murine macrophages, or/and iNOS. MethA tumor-bearing mice were depleted of macrophages. Pre- and post-TNF tumor NO levels were measured continuously, and mice were followed for gross tumor response. In MethA tumors, TNF caused a 96% response rate, and tumor NO concentration doubled. Tumor blood flow decreased to 3% of baseline by 4 hr and was sustained at 24 hr and 10 days post-TNF. Selective NO inhibition with 1400 W blocked NO rise and decreased response rate to 38%. MethA tumors showed tumor infiltration by macrophages post-TNF and the pattern of macrophage immunostaining overlapped with iNOS immunostaining. Depletion of macrophages inhibited tumor NO increase and response to TNF. LL2 tumors had a 0% response rate to TNF and exhibited no change in NO concentration. Blood flow decreased to 2% of baseline by 4 hr, recovered to 56% by 24 hr and increased to 232% by 10 days. LL2 tumors showed no infiltration by macrophages post-TNF. We conclude that TNF causes tumor infiltrating, macrophage-derived iNOS-mediated tumor NO rise and sustained tumor blood flow shutdown, resulting in tumor ulceration in the responsive tumor.


Subject(s)
Antineoplastic Agents/pharmacology , Fibrosarcoma/blood supply , Fibrosarcoma/drug therapy , Macrophages/metabolism , Nitric Oxide/metabolism , Tumor Necrosis Factor-alpha/pharmacology , Animals , Cell Adhesion Molecules/metabolism , Cell Line, Tumor , Clodronic Acid/administration & dosage , Clodronic Acid/pharmacology , E-Selectin/metabolism , Enzyme-Linked Immunosorbent Assay , Female , Fibrosarcoma/enzymology , Fibrosarcoma/metabolism , Gene Expression Regulation, Enzymologic , Immunohistochemistry , Liposomes , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Nitric Oxide Synthase Type II/metabolism , Regional Blood Flow , Vascular Cell Adhesion Molecule-1/metabolism
3.
J Am Acad Dermatol ; 58(1): 68-73, 2008 Jan.
Article in English | MEDLINE | ID: mdl-17692428

ABSTRACT

BACKGROUND: Atopic dermatitis (AD) is often associated with comorbidities such as allergic rhinitis and asthma. OBJECTIVE: We sought to describe the frequency of these comorbidities in children with AD. METHODS: We conducted a cross-sectional study of the first 2270 children with physician-confirmed AD enrolled in a large postmarketing cohort. All were queried for information on comorbidities using a questionnaire from the International Study of Asthma and Allergies in Childhood. RESULTS: In all, 71.3% reported at least one additional form of atopy (symptoms of asthma or allergic rhinitis). A total of 33.3% reported only symptoms of asthma or allergic rhinitis whereas 38.0% reported symptoms of asthma and allergic rhinitis. By age 3 years, nearly 66% reported at least one additional form of atopy. A statistically significant trend toward poorer disease control was observed for those with additional atopic illnesses (P < .001). LIMITATIONS: This is a cross-sectional study. CONCLUSION: Individuals with AD exhibit a predisposition to additional atopic illnesses by age 3 years and in turn the presence of these illnesses correlates with poor disease control.


Subject(s)
Asthma/epidemiology , Dermatitis, Atopic/epidemiology , Hypersensitivity/complications , Rhinitis/epidemiology , Rhinitis/etiology , Adolescent , Asthma/physiopathology , Child , Child, Preschool , Cohort Studies , Comorbidity , Cross-Sectional Studies , Dermatitis, Atopic/diagnosis , Dermatitis, Atopic/physiopathology , Female , Humans , Male , Physicians , Prevalence , Rhinitis/physiopathology
4.
Ann Surg ; 244(5): 781-91, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17060772

ABSTRACT

BACKGROUND: Isolated limb perfusion using high-dose human tumor necrosis factor-alpha with melphalan is effective therapy for bulky extremity in-transit melanoma and sarcoma. OBJECTIVE: While it is widely accepted that melphalan is a DNA alkylating agent, the mechanism of selective antitumor effect of tumor necrosis factor-alpha is unclear. METHODS AND RESULTS: Electron microscopic analyses of human melanoma biopsies, pre- and post-melphalan perfusion, showed that the addition of tumor necrosis factor-alpha caused gapping between endothelial cells by 3 to 6 hours post-treatment followed by vascular erythrostasis in treated tumors. In human melanoma xenografts raised in mice, tumor necrosis factor-alpha selectively increased tumor vascular permeability by 3 hours and decreased tumor blood flow by 6 hours post-treatment relative to treated normal tissue. In an in vitro tumor endothelial cell model, tumor necrosis factor-alpha caused vascular endothelial adherens junction protein, VE-cadherin, to relocalize within the cell membrane away from cell-cell junctions leading to gapping between endothelial cells by 3 to 6 hours post-treatment. Phosphotyrosinylation was a prerequisite for movement of VE-cadherin away from endothelial cell junctions and for gapping between endothelial cells. Clinical isolated limb perfusion tumor specimens, at 3 hours postperfusion, showed a discontinuous and irregular pattern of VE-cadherin expression at endothelial cell junctions when compared with normal (skin) or pretreatment tumor tissue. CONCLUSIONS: Together, the data suggest that tumor necrosis factor-alpha selectively damages the integrity of tumor vasculature by disrupting VE-cadherin complexes at vascular endothelial cell junctions leading to gapping between endothelial cells, causing increased vascular leak and erythrostasis in tumors. VE-cadherin appears to be a potentially good target for selective antitumor therapy.


Subject(s)
Antigens, CD/metabolism , Cadherins/metabolism , Endothelium, Vascular/drug effects , Melanoma, Experimental , Skin Neoplasms , Skin/blood supply , Tumor Necrosis Factor-alpha/therapeutic use , Animals , Antigens, CD/drug effects , Biopsy , Blood Flow Velocity , Cadherins/drug effects , Cell Line, Tumor , Endothelium, Vascular/metabolism , Endothelium, Vascular/ultrastructure , Flow Cytometry , Humans , Melanoma, Experimental/blood supply , Melanoma, Experimental/drug therapy , Melanoma, Experimental/metabolism , Mice , Mice, Nude , Microscopy, Electron , Neoplasm Transplantation , Skin/physiopathology , Skin/ultrastructure , Skin Neoplasms/blood supply , Skin Neoplasms/drug therapy , Skin Neoplasms/metabolism , Transplantation, Heterologous , Treatment Outcome
5.
Clin Cancer Res ; 12(18): 5464-70, 2006 Sep 15.
Article in English | MEDLINE | ID: mdl-17000681

ABSTRACT

PURPOSE: A phase II trial of Photofrin-mediated i.p. photodynamic therapy shown in a previous report limited efficacy and significant acute, but not chronic, toxicity. A secondary aim of this trial and the subject of this report is to determine Photofrin uptake in tumor and normal tissues. EXPERIMENTAL DESIGN: Patients received Photofrin, 2.5 mg/kg, i.v., 48 hours before debulking surgery. Photofrin uptake was measured by spectroflurometric analysis of drug extracted from tumor and normal tissues removed at surgery. Differences in drug uptake among these tissues were statistically considered using mixed-effects models. RESULTS: Photofrin concentration was measured in 301 samples collected from 58 of 100 patients enrolled on the trial. In normal tissues, drug uptake significantly (P<0.0001) differed as a function of seven different tissue types. In the toxicity-limiting tissue of intestine, the model-based mean (SE) Photofrin level was 2.70 ng/mg (0.32 ng/mg) and 3.42 ng/mg (0.24 ng/mg) in full-thickness large and small intestine, respectively. In tumors, drug uptake significantly (P=0.0015) differed as a function of patient cohort: model-based mean Photofrin level was 3.32 to 5.31 ng/mg among patients with ovarian, gastric, or small bowel cancer; 2.09 to 2.45 ng/mg among patients with sarcoma and appendiceal or colon cancer; and 0.93 ng/mg in patients with pseudomyxoma. Ovarian, gastric, and small bowel cancers showed significantly higher Photofrin uptake than full-thickness large and/or small intestine. However, the ratio of mean drug level in tumor versus intestine was modest (

Subject(s)
Dihematoporphyrin Ether/pharmacokinetics , Photochemotherapy/methods , Biopsy , Cohort Studies , Dihematoporphyrin Ether/administration & dosage , Female , Gastrointestinal Neoplasms/drug therapy , Humans , Injections, Intraperitoneal , Models, Biological , Organ Specificity , Ovarian Neoplasms/drug therapy , Sarcoma/drug therapy , Tissue Distribution
6.
Clin Cancer Res ; 12(8): 2517-25, 2006 Apr 15.
Article in English | MEDLINE | ID: mdl-16638861

ABSTRACT

PURPOSE: A previous phase I trial of i.p. photodynamic therapy established the maximally tolerated dose of Photofrin (Axcan Pharma, Birmingham, AL)-mediated photodynamic therapy and showed encouraging efficacy. The primary objectives of this phase II study were to determine the efficacy and toxicities of i.p. photodynamic therapy in patients with peritoneal carcinomatosis and sarcomatosis. EXPERIMENTAL DESIGN: Patients received Photofrin 2.5 mg/kg i.v. 48 hours before debulking surgery. Intraoperative laser light was delivered to the peritoneal surfaces of the abdomen and pelvis. The outcomes of interest were (a) complete response, (b) failure-free survival time, and (c) overall survival time. Photosensitizer levels in tumor and normal tissues were measured. RESULTS: One hundred patients were enrolled into one of three strata (33 ovarian, 37 gastrointestinal, and 30 sarcoma). Twenty-nine patients did not receive light treatment. All 100 patients had progressed by the time of statistical analysis. The median failure-free survival and overall survival by strata were ovarian, 2.1 and 20.1 months; gastrointestinal cancers, 1.8 and 11.1 months; sarcoma, 3.7 and 21.9 months. Substantial fluid shifts were observed postoperatively, and the major toxicities were related to volume overload. Two patients died in the immediate postoperative period from bleeding, sepsis, adult respiratory distress syndrome, and cardiac ischemia. CONCLUSIONS: Intraperitoneal Photofrin-mediated photodynamic therapy is feasible but does not lead to significant objective complete responses or long-term tumor control. Heterogeneity in photosensitizer uptake and tumor oxygenation, lack of tumor specificity for photosensitizer uptake, and the heterogeneity in tissue optical properties may account for the lack of efficacy observed.


Subject(s)
Carcinoma/drug therapy , Dihematoporphyrin Ether/therapeutic use , Peritoneal Neoplasms/drug therapy , Photochemotherapy/methods , Sarcoma/drug therapy , Adult , Antineoplastic Agents/adverse effects , Antineoplastic Agents/therapeutic use , Carcinoma/pathology , Diarrhea/chemically induced , Dihematoporphyrin Ether/adverse effects , Edema/chemically induced , Female , Follow-Up Studies , Humans , Male , Middle Aged , Peritoneal Neoplasms/pathology , Photochemotherapy/adverse effects , Sarcoma/pathology , Sunburn/etiology , Survival Analysis , Treatment Outcome
7.
Cancer Lett ; 221(2): 225-35, 2005 Apr 28.
Article in English | MEDLINE | ID: mdl-15808408

ABSTRACT

Tumor oxygenation status is an independent prognostic indicator in cancer because it influences tumor progression and treatment outcome. Its quantitative value is determined by a number of tumor vascular parameters such as microvascular density, blood flow, blood volume, blood oxygen saturation, tumor tissue pO2, and resistance to oxygen diffusion within the tumor. Over the past several years, considerable time and effort have been invested into developing techniques to effectively and reliably measure the oxygenation status of a tumor. The measurement and interpretation of data obtained with currently available methods is complicated by the heterogeneity in tumor oxygenation. Currently available techniques can be broadly classified into direct invasive methods, direct non-invasive methods, and measurement of surrogate endogenous markers of tumor oxygenation. Of these methods, the Eppendorf pO2 histograph is considered the 'gold standard' and even so has several limitations. Given the importance of tumor oxygenation status in therapy and in predicting disease progression, it is imperative that reliable, globally usable, and technically simplistic methods be developed to yield a consistent, comprehensive, and reliable profile of tumor oxygenation. Until newer more reliable techniques are developed, existing independent techniques or appropriate combinations of techniques should be optimized and validated using known endpoints in tumor oxygenation status and/or treatment outcomes.


Subject(s)
Cell Hypoxia , Neoplasms/diagnosis , Neoplasms/metabolism , Oxygen/metabolism , Animals , Humans , Prognosis
8.
J Clin Endocrinol Metab ; 90(6): 3440-8, 2005 Jun.
Article in English | MEDLINE | ID: mdl-15797967

ABSTRACT

CONTEXT: Currently, there is no effective treatment for iodine-resistant thyroid cancers. OBJECTIVE: As a new approach to treatment, the efficacy of replication-selective, human thyroglobulin (TG) enhancer and promoter-driven, adenovirus (AdhTGEP)-mediated oncolysis was investigated using two well-differentiated thyroid cancer cell lines, XTC (TG positive) and FTC-133 (TG negative), and other control tumor and nontumor cell lines (all TG negative). DESIGN: A cohort study design was used. SETTING: The study setting was laboratory bench-top experiments. SUBJECTS/PARTICIPANTS: In vitro TG-expressing and nonexpressing thyroid cell culture lines, nonthyroid tumor cell lines, as well as preclinical thyroid tumor-bearing mice were studied. INTERVENTION: Adenoviral infection of cell lines was determined by immunohistochemistry, selective replication by one-step growth assays, and cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrozolium (MTS) assay. In vivo tumor growth inhibition was determined by a single intratumoral injection of 1 x 10(9) plaque-forming units AdhTGEP, AdLacZ (control virus), or PBS to 50- to 75-mm(3) tumors. XTC cells showed intense immunohistochemical staining, whereas FTC-133 and all other control cell lines showed minimal staining for viral infection with AdhTGEP. MAIN OUTCOME MEASURES: Cell survival and tumor growth inhibition after adenoviral infection were the main outcome measures. RESULTS: One-step growth assays showed at least a more than 60-fold titer of AdhTGEP in XTC than in FTC-133 cells. Cytotoxicity assays showed approximately 68% cell kill in XTC and minimal cell kill in FTC-133 and all other control cell lines at a multiplicity of infection of 250. There was significant in vivo growth inhibition of AdhTGEP-treated XTC tumors (67 +/- 49 mm(3)) compared with AdLacZ-treated XTC (228 +/- 45 mm(3); P < 0.01), PBS-treated XTC (372 +/- 70 mm(3); P < 0.001), or AdhTGEP-treated FTC-133 tumors (598 +/- 168 mm(3)). CONCLUSION: Replication-selective virus-mediated oncolysis is a potential therapy for recurrent, well-differentiated, TG-secreting thyroid cancer that is unresponsive to standard treatment.


Subject(s)
Adenoviridae/physiology , Enhancer Elements, Genetic , Genetic Therapy , Promoter Regions, Genetic , Thyroglobulin/genetics , Thyroid Neoplasms/therapy , Adenoviridae/genetics , Base Sequence , Cell Line , Cell Survival/drug effects , Cohort Studies , DNA Primers , Humans , Thyroid Neoplasms/pathology , Transfection , Virus Replication
9.
Surgery ; 135(4): 427-36, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15041967

ABSTRACT

BACKGROUND: Interferon-beta (IFNbeta) has multiple antitumor effects; however, its use has been limited by its short half-life in vivo. This limitation may be overcome by IFNbeta gene therapy. We evaluated adenovirus-IFNbeta therapy in an immunocompetent mouse model of carcinomatosis. METHODS: Mice that were treated intraperitoneally 5 days after tumor (mouse ovarian teratoma) inoculation with an adenoviral vector that contains the mouse IFNbeta gene (Ad-IFNbeta), control adenoviral vector or saline solution. Mice were monitored for multiple outcome measures and toxicity. To determine the mechanism of antitumor effect, flow cytometry of ascites fluid was performed to differentiate immune cell populations. Nitric oxide in ascites fluid was measured with an electrochemical microsensor. RESULTS: Tumor burden was decreased and survival was prolonged (P<.001) in the Ad-IFNbeta group after a single treatment of 3.3 x 10(8) plaque-forming units, with acceptable toxicity. By flow cytometry, an increase in the proportion of natural killer cells (from less than 2% of the gated population to more than 8%; P=.024) and an increase in macrophages were seen in the treated animals. Although there was a trend toward increased levels of nitric oxide in Ad-IFNbeta treatment groups, it was not statistically significant. CONCLUSION: IFNbeta gene therapy results in decreased tumor burden and improved survival in an aggressive, immunocompetent mouse model of carcinomatosis. This therapy warrants further evaluation as a treatment for disseminated peritoneal cancer.


Subject(s)
Antineoplastic Agents/therapeutic use , Genetic Therapy/methods , Interferon-beta/therapeutic use , Ovarian Neoplasms/therapy , Teratoma/therapy , Adenoviridae , Animals , Cell Line, Tumor , Female , Genetic Vectors , Immunocompetence , Mice , Mice, Inbred C3H , Models, Animal , Ovarian Neoplasms/pathology , Teratoma/pathology
10.
Cancer Res ; 63(21): 7232-40, 2003 Nov 01.
Article in English | MEDLINE | ID: mdl-14612518

ABSTRACT

Tumor oxygen status is a reliable prognostic marker that impacts malignant progression and outcome of tumor therapy. However, tumor oxygenation is heterogeneous and cannot be sufficiently described by a single parameter. It is influenced by several factors including microvessel density (MVD), blood flow (BF), blood volume (BV), blood oxygen saturation, tissue pO(2), oxygen consumption rate, and hypoxic fraction. The goal of this investigation was to integrate these measurements to obtain a comprehensive profile of tumor oxygenation. Platelet/endothelial cell adhesion molecule immunohistochemistry, the recessed oxygen microelectrode, color and power Doppler ultrasound (DUS), and diffuse light spectroscopy (DLS) were used to measure tumor oxygen status using vascular endothelial growth factor (VEGF)-transfected hypervascular human melanoma xenografts and their nontransfected counterparts as a model. NIH1286 human melanoma cells were transfected with a retroviral vector +/- a 720-bp fragment of human VEGF(121). High VEGF-producing clones were selected by ELISA. Oxygen consumption rate was measured in NIH1286/VEGF+ [VEGF-transfected cells (VEGF+ cells)] and NIH1286/Vec cells [cells transfected with vector alone (Vec cells)] using a standard Clark oxygen electrode. Athymic nude 6-8-week-old mice received s.c. injection in the right flank with 5 x 10(6) VEGF+ or Vec cells. When tumors were 10-14 mm in maximum dimension, serum was analyzed for VEGF by ELISA. Cryopreserved tumor tissue sections were immunostained for platelet/endothelial cell adhesion molecule, and MVD measurements were made. Tumor-bearing mice were anesthetized, and pO(2) measurements were made using Eppendorf pO(2) histograph or the recessed oxygen microelectrode. Tumor BF and BV were measured by quantitative analysis of DUS images. DLS was used to measure tumor BF and blood oxygen saturation variation. VEGF+ cell supernatants had 15,500 pg/ml VEGF, and Vec cells had 10 pg/ml. VEGF+ and Vec cells had equivalent oxygen consumption rates. VEGF+ tumors had a faster growth rate than Vec tumors. Serum from VEGF+ tumor-bearing mice showed 4,211 pg/ml VEGF, whereas VEGF was undetectable in the serum of control mice. MVD values were 74 +/- 11 in VEGF+ tumors and 39 +/- 4 in control tumors at x200 magnification/0.95-mm(2) area. The median pO(2) values were 3.5-fold higher in VEGF+ tumors than in Vec tumors by the recessed oxygen microelectrode and 18-fold higher by Eppendorf pO(2) histograph. DUS showed a 3.3-fold higher mean BF and a 5.5-fold higher BV in VEGF+ tumors than in Vec tumors. DLS showed a 3.2-fold higher mean BF and 1.7-fold higher oxygen saturation in the hypervascular tumors as compared with the control tumor type, consistent with increased BF and BV data by DUS. An integrated approach that yields a comprehensive and consistent profile of oxygen status in tumors could potentially provide critical information for prognosis and treatment.


Subject(s)
Melanoma/blood supply , Oxygen/blood , Animals , Cell Line, Tumor , Humans , Melanoma/diagnostic imaging , Melanoma/metabolism , Mice , Mice, Nude , Neoplasm Transplantation , Oxygen/metabolism , Partial Pressure , Transfection , Transplantation, Heterologous , Ultrasonography, Doppler/methods , Vascular Endothelial Growth Factor A/biosynthesis , Vascular Endothelial Growth Factor A/genetics
11.
Surgery ; 133(4): 420-8, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12717360

ABSTRACT

BACKGROUND: Isolated limb perfusion (ILP) with tumor necrosis factor-alpha (TNF) and melphalan for advanced extremity malignancies achieves significant complete response rates. To study molecular mechanisms underlying this response, a nude rat ILP model with a human melanoma xenograft was developed. METHODS: NIH1286 human melanoma was grown subcutaneously in the hind limb of nude rats. Anesthetized rats underwent a 10-minute ILP via femoral vessels with hetastarch, heparin, and melphalan, TNF, or TNF plus melphalan. The tumors and ulcers were measured and viable tumor area was calculated. Post-ILP tumors were analyzed by electron microscopy for vascular damage and also by liquid chromatography atmospheric pressure chemical ionization tandem mass spectrometry (LC/APCI/MS/MS) for free melphalan levels. Colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-dephenyltetrazolium bromide (MTT) assays were performed on NIH1286 cells and human dermal microvascular endothelial cells (HDMVEC) to test for direct cytotoxicity to TNF and melphalan. Post-ILP tumors sections were also examined by electron microscopy. RESULTS: The mean maximal decrease in viable tumor area after ILP for control, TNF, melphalan, and TNF + melphalan groups were 0%, 22 +/- 13%, 61 +/- 14%, and 100 +/- 0%, respectively. LC/APCI/MS/MS revealed no difference in the free tumor melphalan concentration between melphalan alone and TNF + melphalan groups. The percent cytotoxicity in MTT assays using TNF, melphalan, and TNF + melphalan against NIH1286 were 0%, 51-59%, and 74-81%, respectively, and against HDMVEC were 28%, 16-23%, and 6-13%, respectively. Electron microscopic analyses showed that addition of TNF to the perfusate caused erythrostasis in tumor blood vessels. CONCLUSION: We developed a human melanoma nude rat ILP model with tumor responses very similar to the human ILP trials. This model will allow further investigation of the synergistic mechanism of TNF and melphalan in human melanoma in a preclinical setting, and extension of this study to current clinical trials.


Subject(s)
Antineoplastic Agents, Alkylating/pharmacology , Antineoplastic Agents/pharmacology , Melanoma/drug therapy , Melphalan/pharmacology , Skin Neoplasms/drug therapy , Tumor Necrosis Factor-alpha/pharmacology , Animals , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Disease Models, Animal , Female , Hindlimb , Humans , Perfusion , Rats , Rats, Nude , Tumor Cells, Cultured , Xenograft Model Antitumor Assays
12.
Am J Physiol Heart Circ Physiol ; 284(1): H317-29, 2003 Jan.
Article in English | MEDLINE | ID: mdl-12388254

ABSTRACT

High-dose TNF with melphalan has significant antitumor activity in regional perfusion of the limbs and liver in human malignancies. TNF is believed to target tumor vasculature, but the precise molecular mechanism is unknown. The present study demonstrates that TNF downregulates the VEGF receptor, fetal liver kinase-1 (Flk-1), on tumor endothelium in a human melanoma xenograft model. NIH1286 human melanoma cells were transduced with a 720-bp fragment of the human VEGF(121) gene to develop well-vascularized tumors that served as an amplified system for measuring Flk-1 expression changes. We injected 5 x 10(6) cells subcutaneously, each of two distinct single cell clones (NIH1286/3 and NIH1286/15), into athymic nude mice to produce tumors approximately 10 mm in size. Each animal then received either BSA or TNF in BSA by tail vein. Tumors harvested at different time points post-TNF were analyzed for Flk-1 mRNA and protein expression. Data obtained showed that intravascular TNF downregulated Flk-1 expression in tumor endothelial cells. This effect could contribute to the antitumor activity of TNF known to target tumor vasculature.


Subject(s)
Melanoma/metabolism , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor Receptor-2/metabolism , Animals , Down-Regulation , Endothelial Growth Factors/genetics , Gene Expression , Humans , Intercellular Signaling Peptides and Proteins/genetics , Lymphokines/genetics , Melanoma/genetics , Mice , Mice, Nude , Neoplasm Transplantation , RNA, Messenger/metabolism , Serum Albumin, Bovine/pharmacology , Transplantation, Heterologous , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/pharmacology , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factor Receptor-2/genetics , Vascular Endothelial Growth Factors
13.
Surgery ; 132(2): 252-8, 2002 Aug.
Article in English | MEDLINE | ID: mdl-12219020

ABSTRACT

BACKGROUND: Isolated limb perfusion (ILP) with melphalan is an accepted treatment for intransit melanoma of the extremities. Using an ILP human melanoma xenograft model, we tested the hypothesis that acidosis augments the antitumor effect of melphalan and that nitric oxide (NO) induction mediates tumor regression. METHODS: NIH1286 human melanoma tumor bearing athymic nude rats underwent a 10-minute ILP. Group C was perfused at physiologic pH without acid or melphalan, group M received melphalan at physiologic pH (7.2), group A received 0.2 N of HCl at pH 6.8, and group A/M received melphalan and HCl at pH 6.8. Groups 1400W + A and 1400W + A/M were injected with 1400W, a specific inhibitor of inducible NO synthase, 1 hour pre-ILP. Tumor response was followed for up to 60 days in all survival experiments. In 4 to 6 animals from groups C, M, A, and A/M, tumor NO was measured pre- and post-ILP, and tumor and thigh muscle from 2 additional animals in each group were collected at 20 minutes and 24 hours post-ILP and processed for terminal deoxynucleotidyl transferase dUTP nick end labeling staining. RESULTS: Maximum mean reduction in tumor size after ILP in the different groups was as follows: C = 0%, M = 55%, A = 99.6% (3 of 4 complete responses), A/M = 100% (all complete responses), 1400W + A = 0%, and 1400W + A/M = 25%. Median tumor NO was 0.87 +/- 0.74 (SD) micromol/L before ILP and increased significantly (Mann-Whitney rank sum test, P <.001) after ILP (C = +6.9%, n = 4; M = +7.5%, n = 5; A = +66.0%, n = 6; A/M = +35.9%, n = 6). Also, minimal apoptotic cell death was seen in C and M, whereas A and A/M showed evidence of widespread apoptosis. CONCLUSIONS: Acidosis enhances the antitumor effect of melphalan. NO induction appears to play a role in tumor regression.


Subject(s)
Acidosis/metabolism , Antineoplastic Agents, Alkylating/pharmacology , Melanoma/drug therapy , Melphalan/pharmacology , Skin Neoplasms/drug therapy , Amidines/pharmacology , Animals , Apoptosis/drug effects , Apoptosis/physiology , Benzylamines/pharmacology , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Female , Hindlimb/blood supply , Humans , Melanoma/metabolism , Nitric Oxide/metabolism , Rats , Rats, Nude , Regional Blood Flow , Skin Neoplasms/metabolism , Tumor Cells, Cultured/transplantation , Xenograft Model Antitumor Assays
14.
Ann Surg Oncol ; 9(4): 411-8, 2002 May.
Article in English | MEDLINE | ID: mdl-11986195

ABSTRACT

BACKGROUND: Mature dendritic cells (DCs) are potent antigen-presenting cells that activate naive T lymphocytes and initiate cellular immune responses. The ability of CD83(+) mature DCs infected with vaccinia virus encoding the gp100 melanoma transgene (rV-gp100) to stimulate an antimelanoma CD8(+) T-cell response was investigated. METHODS: Monocyte-derived immature or CD83(+) mature DCs were infected with rV-gp100. The activation state of the DCs and the expression of gp100 protein were evaluated by flow cytometry. The reactivity of antimelanoma CD8(+) T cells was confirmed by measuring specific interferon gamma secretion by using enzyme-linked immunosorbent assay in a mixed-tumor lymphocyte culture. RESULTS: Both immature and CD83(+) mature DCs expressed gp100 protein when the DCs were infected with rV-gp100. Calcium-signaling agents were required to induce maturation of both infected and noninfected immature DCs. Only rV-gp100-infected CD83(+) DCs induced CD8(+) T cells, after a single stimulation that recognized both peptide-pulsed target cells to multiple gp100 epitopes and a melanoma cell line that endogenously expressed gp100 antigen. CONCLUSIONS: CD83(+) DCs transduced with rV-gp100 are capable of generating a strong CD8(+) T-cell response against melanoma tumor cells. Expression of melanoma antigens by mature DCs offers the potential advantage of presenting multiple endogenously processed T-cell epitopes and using multiple HLA restriction elements for antimelanoma vaccine therapy.


Subject(s)
Antigen Presentation/genetics , HLA-A2 Antigen/genetics , Immunoglobulins/genetics , Melanoma/immunology , Membrane Glycoproteins/genetics , Neoplasm Proteins/genetics , Vaccinia virus/genetics , Antigens, CD , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/metabolism , Cell Line , Cytotoxicity, Immunologic/genetics , Dendritic Cells/immunology , Dendritic Cells/metabolism , Flow Cytometry , HLA-A2 Antigen/metabolism , Humans , Immunoglobulins/immunology , Membrane Glycoproteins/immunology , Neoplasm Proteins/immunology , Tumor Cells, Cultured , Vaccinia virus/immunology , gp100 Melanoma Antigen , CD83 Antigen
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