ABSTRACT
The egg yolk precursor protein, vitellogenin (Vg), was isolated by size exclusion and ion exchange chromatography from plasma of California halibut (Paralichthys californicus) treated with estrogen. MALDI TOF mass spectrometry (MS) analysis resulted in a molecular mass of 188 kDa. MS/MS de novo sequencing identified the protein as Vg by matching sequences of tryptic peptides to the known sequences of several other species. Matches were also made to two different forms of Vg in haddock, medaka, and mummichog, providing evidence that California halibut has more than one form of Vg. Native PAGE and Western blot with an antibody to turbot (Scophthalmus maximus) Vg confirmed the identity of the protein. Protein resolved on the SDS PAGE as a double band of approximately the same mass as determined with MALDI TOF, and two lower mass bands that were also immunoreactive. MALDI TOF and MS/MS de novo sequencing were useful for determining the molecular mass, identification, and exploring the multiplicity of Vg. The potential of using other MS methods to understand the structure and function of Vg is discussed.
Subject(s)
Flounder/physiology , Vitellogenins/biosynthesis , Amino Acid Sequence , Animals , Blotting, Western , Electrophoresis, Polyacrylamide Gel , Estradiol/pharmacology , Male , Molecular Sequence Data , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tandem Mass Spectrometry , Vitellogenins/chemistryABSTRACT
AIMS: Immunohistochemistry (IHC) and fluorescence in-situ hybridization (FISH) are both commonly used assays for evaluation of HER-2/neu status in breast cancer. However, there is still no consensus on which method is most predictive of patient response to Herceptin. Recently, the automated cellular imaging system (ACIS) has been shown to improve the accuracy and reproducibility in scoring IHC. Our aim was to compare the results of HER-2/neu expression and gene amplification in the same patients by IHC using the ACIS system and by FISH. METHODS AND RESULTS: Two hundred and forty-seven breast cancer cases were studied. The concordance rate between IHC-ACIS (> or = 2.2) and FISH (> or = 2.0) was 94%. Fifteen patients were discordant; three had borderline FISH values and three had borderline IHC values. The other nine discordant cases consisted of five IHC-ACIS+, FISH- and six IHC-ACIS-, FISH+. HER-2/neu overexpression was more common in tumours that were high-grade, aneuploid, progesterone receptor and bcl-2 negative, with MIB-1 > 10%. CONCLUSION: HER-2/neu assessment by the ACIS is reliable, rapid and inexpensive, and correlates highly with results obtained by FISH.
Subject(s)
Breast Neoplasms/chemistry , Image Processing, Computer-Assisted/methods , Immunohistochemistry/methods , In Situ Hybridization, Fluorescence , Receptor, ErbB-2/analysis , Receptor, ErbB-2/genetics , Adenocarcinoma/chemistry , Adenocarcinoma/genetics , Adenocarcinoma, Mucinous/chemistry , Adenocarcinoma, Mucinous/genetics , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Breast Neoplasms/genetics , Carcinoma, Ductal, Breast/chemistry , Carcinoma, Ductal, Breast/genetics , Carcinoma, Lobular/chemistry , Carcinoma, Lobular/genetics , Carcinoma, Medullary/chemistry , Carcinoma, Medullary/genetics , Carcinoma, Squamous Cell/chemistry , Carcinoma, Squamous Cell/genetics , Female , Gene Expression Regulation, Neoplastic , Humans , Middle Aged , Receptor, ErbB-2/biosynthesisABSTRACT
Sex and age influence morphine analgesia in humans and animals. Mature rats show greater morphine analgesia in males than in females. Ultra-low doses of naltrexone enhance morphine analgesia. In mature rats (18-22 weeks), naltrexone (0.002-2.0 mg/kg)-morphine (2 mg/kg) cotreatment enhanced morphine analgesia in females, an effect inversely related to naltrexone dose. Conversely, in mature male rats, naltrexone tended to decrease morphine analgesia with increasing dose. In young rats (8-10 weeks), morphine analgesia was unrelated to sex and in both sexes the naltrexone-morphine interaction was negligible. These data show that dose, age, and sex alter the naltrexone-morphine interaction in rats.
Subject(s)
Morphine/administration & dosage , Naltrexone/administration & dosage , Age Factors , Analgesics, Opioid/administration & dosage , Animals , Drug Interactions , Female , Male , Narcotic Antagonists/administration & dosage , Narcotics/administration & dosage , Pain Threshold/drug effects , Rats , Rats, Sprague-Dawley , Receptors, Opioid/drug effects , Sex FactorsABSTRACT
Several studies indicate greater sensitivity to morphine (MOR) analgesia in male compared to female rats under the acute dosing condition. The present study investigated whether the same sex difference in sensitivity persists in MOR-tolerant rats. MOR was administered chronically (7 mg/kg twice daily) until tolerance developed in each rat. Tolerant rats were treated randomly with higher graded doses of MOR (10-25 mg/kg). Analgesia (tail-flick test) and spontaneous motor activity (total locomotion) were measured. The present data confirmed previous studies showing a greater sensitivity to acute MOR in male than in female rats. However, the sex differences seen in MOR sensitivity were abolished in tolerant rats. The rate of acquisition of tolerance was similar in male and female rats. The analgesic response was not affected by motor depression.
Subject(s)
Drug Tolerance/physiology , Morphine/pharmacology , Sex Characteristics , Animals , Dose-Response Relationship, Drug , Female , Male , Motor Activity/drug effects , Motor Activity/physiology , Pain Measurement/drug effects , Pain Measurement/methods , Rats , Rats, Sprague-DawleyABSTRACT
The 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazol[1,5-a][1,4]benzodiazepine-3-carboxylic acid ethyl ester (Flumazenil)-morphine interaction on analgesia (acute pain model, tail-flick test) was tested after intraperitoneal (IP) and intrathecal (IT) routes of administration in female rats. Analgesia was enhanced by the concurrent administration of Flumazenil with morphine (IP), in a dose-related way. Flumazenil alone (IP) did not produce analgesia. In contrast, morphine analgesia was not enhanced by Flumazenil by the IT route. These data demonstrate that Flumazenil enhances morphine-mediated antinociception by mechanisms that are likely to involve benzodiazepine receptors at sites other than the spinal cord.
Subject(s)
Analgesics, Opioid/pharmacology , Flumazenil/pharmacology , Morphine/pharmacology , Animals , Benzodiazepines/antagonists & inhibitors , Drug Interactions , Female , Flumazenil/administration & dosage , Rats , Rats, Sprague-DawleyABSTRACT
This study assessed the effect of the central benzodiazepine receptor antagonist, 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3-carboxylic acid ethyl ester (flumazenil), on morphine-induced analgesia, locomotor effects, and development of tolerance in rats. The thermally evoked pain (tail flick) response was determined after acute and chronic intraperitoneal (i.p.) administration of morphine and flumazenil, alone and in combination. In acute studies, flumazenil induced weak analgesia unrelated to dose and sex, whereas morphine-induced analgesia was dependent on both dose and sex (male>female). Flumazenil dose-dependently enhanced the analgesic effect of morphine in female but not in male rats. Isobolographic analysis suggested synergism between flumazenil and morphine in female rats, but antagonism in male rats. Flumazenil-induced locomotor changes (alone and with morphine) were related to sex but not dose. Chronic coadministration of flumazenil with morphine enhanced analgesia and attenuated tolerance development in female rats. The findings suggest a possible role for flumazenil as an adjunct with opioids in acute and chronic pain therapy.
Subject(s)
Analgesia/methods , Drug Tolerance/physiology , Flumazenil/pharmacology , Morphine/pharmacology , Sex Characteristics , Animals , Dose-Response Relationship, Drug , Drug Interactions/physiology , Drug Therapy, Combination , Female , Locomotion/drug effects , Locomotion/physiology , Male , Pain Measurement/drug effects , Pain Measurement/methods , Rats , Rats, Sprague-DawleyABSTRACT
BACKGROUND: Widespread screening mammography has resulted in detection of many breast cancers smaller than one cm. Image-guided percutaneous needle sampling provides accurate diagnostic and prognostic information for adjuvant therapy. Less invasive methods based on imaging techniques are emerging as an alternative to wire localization and lumpectomy. DATA SOURCES: Information presented in this overview was provided by seven investigators from five medical centers in the United States. These researchers are currently developing various techniques of image-guided percutaneous therapy of small (Tis, 1) breast cancers. CONCLUSIONS: Several percutaneous treatment modalities for treatment of early breast cancer, either excisional or in-situ ablative, are described in this overview and their potential applications are discussed.
Subject(s)
Breast Neoplasms/pathology , Breast Neoplasms/surgery , Biopsy , Catheter Ablation , Female , Humans , Laser Therapy , Stereotaxic TechniquesABSTRACT
The objective of this study was to determine the safety and antitumor activity of an autologous GM-CSF-secreting melanoma cell vaccine that was engineered ex vivo with recombinant replication-incompetent adenovirus harboring a human GM-CSF gene (Adv/hGM-CSF). Melanoma samples were surgically obtained from 30 patients (15 female and 15 male, ages ranging from 23 to 87) and were processed for vaccine preparation. Due to stringent eligibility criteria, 9 out of 30 patients were enrolled in the phase 1 clinical trial (FDA IND7677). Melanoma cell lines established from surgical specimens of 9 patients were transduced with Adv/hGM-CSF (MOI of 100) and subsequently irradiated at 35 Gy. These cell lines secreted human GM-CSF in vitro at an average rate of 80-424 ng/10(6) cells/24 h. All patients were intradermally and subcutaneously injected at several sites with irradiated autologous melanoma cells (2x10(6)-1x10(7) in 300 microl saline), 2-10 times, at intervals of 4-8 weeks. None of the patients vaccinated showed any serious adverse systemic response. Three patients (nos.1, 6 and 7) demonstrated local reaction (erythema) to the vaccination. Tumor-specific CTL assays performed in the absence of K562 cells showed that the levels of CTLs in peripheral blood of 5 patients increased following vaccination, whereas those in one patient declined. Levels of CTLs assayed in the presence of K562 cells were considerably lower than those assayed in the absence of K562 cells, but were also found to increase following vaccination in the peripheral blood of 6 patients. A patient who had been vaccinated 10 times (patient 1) responded to the vaccination by apparent reduction in size of metastatic tumor in the lung. Immunohistochemical examination of the vaccination sites of patient 1, biopsied after the 3rd and 4th vaccination. showed that the vaccination sites responded with infiltration of inflammatory cells, such as T cells (CD3+, CD8+), macrophages and dendritic cells (CD83+), for a period up to about 8 days. These data suggest that repeated vaccinations with irradiated autologous GM-CSF-producing tumor cells were well tolerated by patients and led to the activation of an antitumor immune response in some patients.
Subject(s)
Adenoviridae/genetics , Cancer Vaccines/immunology , Genetic Therapy , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Melanoma/therapy , Adult , Aged , Aged, 80 and over , Female , Humans , Hypersensitivity, Delayed , Male , Melanoma/immunology , Middle Aged , T-Lymphocytes, Cytotoxic/immunology , Transduction, Genetic , VaccinationABSTRACT
Consumption of tomato products has been associated with decreased risk of some cancer types, and the tomato antioxidant, lycopene, is thought to play an important role in the observed health effects. In this study, four carotenoids, trans-lycopene, phytofluene, phytoene, and zeta-carotene, were quantified in tomato products. Samples of raw tomatoes, tomato juice after hot break scalder, and final paste were obtained from two different processing plants over two years. Comparison of carotenoid levels throughout processing indicated that lycopene losses during processing of tomatoes into final paste (25-30 degrees Brix) ranged from 9 to 28%. The initial Brix level of the raw tomatoes appeared to influence the amount of lycopene loss that occurred, possibly due to the differences in processing time required to achieve the final desired Brix level of the paste. In general, no consistent changes in the other carotenoids were observed as a function of processing. The antioxidant activity of fresh tomatoes, tomato paste, and three fractions obtained from these products (i.e., aqueous, methanol, and hexane fractions) was also determined. In both a free radical quenching assay and a singlet oxygen quenching assay, significant antioxidant activity was found in both the hexane fraction (containing lycopene) and the methanol fraction, which contained the phenolic antioxidants caffeic and chlorogenic acid. The results suggest that in addition to lycopene, polyphenols in tomatoes may also be important in conferring protective antioxidative effects.
Subject(s)
Antioxidants/analysis , Carotenoids/analysis , Food Handling/methods , Solanum lycopersicum/chemistry , Antioxidants/metabolism , Lycopene , Time FactorsABSTRACT
BACKGROUND: Treatment-related factors that influence quality of life (QOL) for women who are diagnosed with breast carcinoma require study. This study was designed to examine the convergent validity of objective and subjective indices of cosmetic and functional status after patients undergo breast-conserving treatment (BCT) and to test the relations of the objective indicators with QOL. METHODS: Women (n = 54 patients) who had received BCT and radiotherapy for Stage 0-II disease for whom the diagnosis duration ranged from 9 months to 18 years completed assessments of background information, perceived cosmetic and functional outcomes, and QOL. They also were administered a clinical breast examination, including objective ratings of arm edema and breast cosmesis. The patients were a subsample from the study reported by the authors in an accompanying article that is presented in this issue. RESULTS: The findings revealed positive cosmetic and functional treatment outcomes, such that 82% of patients had no or minimal arm edema, and 70% of patients evidenced good or excellent cosmetic results. Convergent validity was demonstrated for the objective and subjective assessments of cosmetic and functional treatment outcomes. In addition, women who had more arm edema reported poorer QOL with regard to depressive symptoms (P < 0.05) and fear of disease recurrence (P < 0.05). CONCLUSIONS: The findings from this study and those reported in the accompanying article reveal that functional treatment outcomes, such as arm edema and breast specific pain, are important correlates of QOL even many years after patients undergo BCT and radiotherapy. Both subjective and objective indicators of treatment outcomes are useful predictors of QOL.
Subject(s)
Breast Neoplasms/therapy , Mastectomy, Segmental , Quality of Life , Breast Neoplasms/psychology , Breast Neoplasms/radiotherapy , Breast Neoplasms/surgery , Combined Modality Therapy , Depression/etiology , Esthetics , Female , Humans , Middle Aged , Treatment OutcomeABSTRACT
Radiotherapy after breast-conserving surgery increases local control. We tested the feasibility of limited surgery with tumor bed irradiation only with 192Ir in a selected group of patients with stage I breast cancer. Twenty-five breasts in 24 women more than 60 years old with low- or intermediate-grade stage I tumors were treated with placement of interstitial catheters at the time of lumpectomy and axillary node dissection. This procedure was followed by after-loading with low-dose 192Ir to deliver 20-25 Gy to the tumor bed over 24-48 hours. There were neither local recurrences in the breast nor distant recurrences at a median follow-up of 47 months (range 25-90 months). Cosmetic appearance ranged from very good to excellent. There were no long-term complications. It is feasible to treat a select group of patients with tumor bed irradiation, using relatively low doses of interstitial irradiation, with excellent local control and no significant morbidity.
Subject(s)
Breast Neoplasms/radiotherapy , Breast Neoplasms/surgery , Carcinoma, Intraductal, Noninfiltrating/radiotherapy , Carcinoma, Intraductal, Noninfiltrating/surgery , Aged , Aged, 80 and over , Brachytherapy , Breast Neoplasms/pathology , Carcinoma, Intraductal, Noninfiltrating/pathology , Disease-Free Survival , Female , Humans , Iridium Radioisotopes/therapeutic use , Mastectomy, Segmental , Middle Aged , Neoplasm Staging , Pilot Projects , Treatment OutcomeABSTRACT
BACKGROUND: The Intergroup Melanoma Surgical Trial began in 1983 to examine the optimal surgical margins of excision for primary melanomas of intermediate thickness (i.e., 1-4 mm). There is now a median 10-year follow-up. METHODS: There were two cohorts entered into a prospective multi-institutional trial: (1) 468 patients with melanomas on the trunk or proximal extremity who randomly received a 2 cm or 4 cm radial excision margin and (2) 272 patients with melanomas on the head, neck, or distal extremities who received a 2 cm radial excision margin. RESULTS: A local recurrence (LR) was associated with a high mortality rate, with a 5-year survival rate of only 9% (as a first relapse) or 11% (anytime) compared with an 86% survival for those patients who did not have a LR (P < .0001). The 10-year survival for all patients with a LR was 5%. The 10-year survival rates were not significantly different when comparing 2 cm vs. 4 cm margins of excision (70% vs. 77%) or comparing the management of the regional lymph nodes (observation vs. elective node dissection). The incidences of LR were the same for patients having a 2 cm vs. 4 cm excision margin regardless of whether the comparisons were made as first relapse (0.4% vs. 0.9%) or at anytime (2.1% vs. 2.6%). When analyzed by anatomic site, the LR rates were 1.1% for melanomas arising on the proximal extremity, 3.1% for the trunk, 5.3% for the distal extremities, and 9.4% for the head and neck. The most profound influence on LR rates was the presence or absence of ulceration; it was 6.6% vs. 1.1% in the randomized group involving the trunk and proximal extremity and was 16.2% vs. 2.1% in the non-randomized group involving the distal extremity and head and neck (P < .001). A multivariate (Cox) regression analysis showed that ulceration was an adverse and independent factor (P = .0001) as was head and neck melanoma site (P = .01), while the remaining factors were not significant (all with P > .12). CONCLUSION: For this group of melanoma patients, a local recurrence is associated with a high mortality rate, a 2-cm margin of excision is safe and ulceration of the primary melanoma is the most significant prognostic factor heralding an increased risk for a local recurrence.
Subject(s)
Melanoma/surgery , Skin Neoplasms/surgery , Humans , Lymph Node Excision/adverse effects , Melanoma/mortality , Melanoma/pathology , Multivariate Analysis , Neoplasm Invasiveness/pathology , Neoplasm Recurrence, Local/mortality , Neoplasm Staging , Neoplasm, Residual , Prospective Studies , Regression Analysis , Skin Neoplasms/mortality , Skin Neoplasms/pathology , Survival Rate , Time Factors , Treatment OutcomeABSTRACT
BACKGROUND: Ten- to 15-year survival results were analyzed from a prospective multi-institutional randomized surgical trial that involved 740 stages I and II melanoma patients with intermediate thickness melanomas (1.0 to 4.0 mm) and compared elective (immediate) lymph node dissection (ELND) with clinical observation of the lymph nodes as well as prognostic factors that independently predict outcomes. METHODS: Eligible patients were stratified according to tumor thickness, anatomical site, and ulceration, and then prerandomized to either ELND or nodal observation. By using Cox stepwise multivariate regression analysis, the independent predictors of outcome were tumor thickness (P < .001), the presence of tumor ulceration (P < .001), trunk site (P = .003), and patient age more than 60 years (P = .01). RESULTS: Overall 10-year survival was not significantly different for patients who received ELND or nodal observation (77% vs. 73%; P = .12). Among the prospectively stratified subgroups of patients, 10-year survival rates favored those patients with ELND, with a 30% reduction in mortality rate for the 543 patients with nonulcerated melanomas (84% vs. 77%; P = .03), a 30% reduction in mortality rate for the 446 patients with tumor thickness of 1.0 to 2.0 mm (86% vs. 80%; P = .03), and a 27% reduction in mortality rate for 385 patients with limb melanomas (84% vs. 78%; P = .05). Of these subgroups, the presence or absence of ulceration should be the key factor for making treatment recommendations with regard to ELND for patients with intermediate thickness melanomas. CONCLUSIONS: These long-term survival rates from patients treated at 77 institutions demonstrate that ulceration and tumor thickness are dominant predictive factors that should be used in the staging of stages I and II melanomas, and confer a survival advantage for these subgroups of prospectively defined melanoma patients.
Subject(s)
Lymph Node Excision , Melanoma/mortality , Melanoma/surgery , Skin Neoplasms/mortality , Skin Neoplasms/surgery , Extremities , Head and Neck Neoplasms/mortality , Head and Neck Neoplasms/pathology , Head and Neck Neoplasms/surgery , Humans , Melanoma/pathology , Middle Aged , Neoplasm Staging , Prognosis , Proportional Hazards Models , Prospective Studies , Skin Neoplasms/pathology , Treatment OutcomeABSTRACT
Molinate undergoes oxidative metabolism forming either ring-hydroxylated metabolites or molinate sulfoxide. Our previous studies strongly implicated the sulfoxidation pathway in molinate-induced testicular toxicity. The present study compares the metabolic capability of rat and human liver microsomes and slices to form either nontoxic ring-hydroxylated metabolites of molinate or the toxic metabolites derived from the sulfoxidation of molinate. Km and Vmax values indicate that sulfoxidation would be the preferred high-dose pathway whereas hydroxylation would predominate at low dose levels in both species. Examination of phase II metabolism of molinate in liver slices reveals greater detoxification of molinate sulfoxide by glutathione conjugation in humans with rats forming less conjugate. Oxidative metabolism of molinate in both rats and humans appears to be mediated by cytochrome P-450 and not flavin monooxygenases as indicated by the use of metabolic inhibitors. Overall, the metabolism of molinate would be via the nontoxic hydroxylation pathway in both species at low doses whereas at high doses, where sulfoxidation would predominate, the human is more capable than the rat to detoxify via glutathione conjugation.
Subject(s)
Azepines/pharmacokinetics , Carbamates , Herbicides/pharmacokinetics , Microsomes, Liver/metabolism , Thiocarbamates , Animals , Cytochrome P-450 Enzyme System/metabolism , Humans , In Vitro Techniques , Microsomes, Liver/enzymology , Oxygenases/metabolism , RatsABSTRACT
Molinate is a thiocarbamate herbicide widely used in rice culture. Studies conducted for regulatory purposes have indicated that molinate exposure causes male reproductive damage in rats. The present study describes the testicular lesion after administration of single doses of molinate. The hypothesis that a metabolite of molinate is responsible for testicular toxicity was also investigated. Testicular damage was evaluated histopathologically in Sprague-Dawley rats 48 h and 1, 2, and 3 weeks after administration of molinate (100-400 mg/kg i.p.). No testicular damage was seen at any time point at the 100 mg/kg dose level. Damage was first seen 1 week after 200 mg/kg and 48 h after 400 mg/kg. The lesion was characterized by Sertoli cell vacuolation, failed spermiation, and phagocytosis of spermatids particularly evident at Stages X and XI. With increasing time, damage progressed until disorganization of the seminiferous epithelium was extensive, multinucleated giant cells were numerous, and neither spermatozoa nor late step spermatids were present. At 3 weeks after administration of the two higher-dose levels, germ cells in the seminiferous tubules were almost completely absent. Administration of the sulfoxide metabolite of molinate (200 mg/kg i.p.) caused testicular damage similar in severity to that seen at the 400 mg/kg dose level for the parent compound, indicating that it was more potent as a testicular toxicant. In vitro metabolism studies using liver and testis microsomes found that the major metabolite in both preparations was molinate sulfoxide. Testis microsomes produced only slightly less sulfoxide when compared with liver microsomes. Molinate was also metabolized via ring hydroxylation to form small amounts of hydroxymolinate. The amount of hydroxymolinate was substantially less in testis microsomes. Overall, these data indicate that sulfoxidation of molinate plays a role in molinat-induced testicular toxicity. Moreover, molinate is metabolized readily by both liver and testis microsomal enzymes, suggesting that the molinate toxic metabolite could be formed in the testis in close proximity to its site of action.
Subject(s)
Azepines/toxicity , Carbamates , Herbicides/toxicity , Pesticide Synergists/toxicity , Safrole/analogs & derivatives , Testis/drug effects , Thiocarbamates , Animals , Azepines/administration & dosage , Azepines/pharmacokinetics , Biotransformation , Dose-Response Relationship, Drug , Giant Cells/drug effects , Herbicides/administration & dosage , Herbicides/pharmacokinetics , In Vitro Techniques , Male , Pesticide Synergists/administration & dosage , Pesticide Synergists/pharmacokinetics , Phagocytosis/drug effects , Rats , Rats, Sprague-Dawley , Safrole/administration & dosage , Safrole/pharmacokinetics , Safrole/toxicity , Seminiferous Tubules/drug effects , Seminiferous Tubules/pathology , Sertoli Cells/drug effects , Sertoli Cells/pathology , Spermatids/drug effects , Spermatids/pathology , Spermatogenesis/drug effects , Testis/pathology , Time FactorsABSTRACT
Molinate, a herbicide widely used on rice, has been previously shown to cause testicular toxicity when a single dose is administered to Sprague-Dawley rats. The sulfoxide metabolite of molinate also was capable of eliciting testicular damage but at lower dose levels than molinate, suggesting that metabolic activation via sulfoxidation could be important in testicular toxicity. Both the sulfoxide and sulfone metabolites of molinate are electrophilic and molinate covalent binding to cellular macromolecules has been attributed to formation of these reactive metabolites. The present study has investigated the nature of the binding reaction of 14C-molinate as well as 14C-molinate sulfoxide and 14C-molinate sulfone in liver and testis microsomal preparations. All three compounds in preparations from both tissues bound extensively and tightly to only one protein of approximately 60 kDa molecular weight on SDS-PAGE. Isoelectric focusing PAGE revealed a pI of approximately 6.0 and native PAGE analysis revealed a native molecular weight of 180 kDa. These data, along with the ability of phenylmethylsulfonyl fluoride to block binding of the 14C-molinate, suggested the molinate-bound protein was an esterase. The protein was purified to homogeneity and MALDI-TOF mass spectral analysis was consistent with Hydrolase A, a carboxylesterase present in both liver and testis. N-terminal sequence analysis revealed 100% homology with Hydrolase A for the first 17 residues. The effect of molinate administration on in vivo esterase activity was assessed both by enzymatic measurement and by histochemical measurement. Molinate treatment caused a marked inhibition of nonspecific esterase activity in both liver and testis. In the testis, histochemical staining showed the esterase activity inhibited by molinate was localized primarily to the Leydig cell, consistent with the localization of Hydrolase A. From these data, it is proposed that molinate-induced inhibition of esterase activity in the Leydig cell could inhibit the mobilization of cholesterol esters required for testosterone biosynthesis.
Subject(s)
Azepines/metabolism , Carbamates , Carboxylic Ester Hydrolases/metabolism , Herbicides/metabolism , Pesticide Synergists/metabolism , Safrole/analogs & derivatives , Thiocarbamates , Amino Acid Sequence , Animals , Carboxylesterase , Carboxylic Ester Hydrolases/analysis , Leydig Cells/drug effects , Leydig Cells/enzymology , Male , Microsomes, Liver/drug effects , Microsomes, Liver/enzymology , Molecular Sequence Data , Rats , Rats, Sprague-Dawley , Safrole/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Sulfones/metabolism , Testis/drug effects , Testis/enzymologyABSTRACT
BACKGROUND: Transhiatal esophagectomy is a popular method of resection because of its reported lower morbidity and mortality and similar survival rates compared to transthoracic esophagectomy. A review of recent experience with these two procedures for resection of distal esophageal and cardia adenocarcinoma is reported. METHODS: From 1988 to 1994, 48 patients with adenocarcinoma of the distal esophagus and gastric cardia were resected with intent to cure, 32 by transhiatal esophagectomy (group 1) and 16 by transthoracic esophagectomy (group II). The two groups were comparable in terms of patient demographics, preoperative risk factors, tumor stage, tumor differentiation, and involvement of resection margins (all not significant [NS]). RESULTS: There was no significant difference in median intensive care unit stay, median hospital stay, incidence of postoperative anastomotic leak, and stricture. Respiratory complications were higher in group I (41% versus 6%, P = 0.01). Hospital mortality was not significantly different for the two groups (group I 3.1% versus group II 0%, NS). Actuarial 5-year survival rates (Kaplan-Meier) were 12% for group I and 39% for group II (NS). CONCLUSIONS: These results suggest that when compared with transhiatal esophagectomy, the transthoracic approach is at least as safe, has comparable complication and survival rates, and remains an acceptable procedure for resection of adenocarcinomas of the distal esophagus and gastric cardia.
Subject(s)
Adenocarcinoma/surgery , Esophageal Neoplasms/surgery , Esophagectomy/methods , Stomach Neoplasms/surgery , Adenocarcinoma/mortality , Adolescent , Adult , Cardia , Esophageal Neoplasms/mortality , Female , Humans , Male , Middle Aged , Postoperative Complications/epidemiology , Stomach Neoplasms/mortality , Survival Rate , ThoraxABSTRACT
OBJECTIVE: A prospective multi-institutional randomized surgical trial involving 740 stage I and II melanoma patients was conducted by the Intergroup Melanoma Surgical Program to determine whether elective (immediate) lymph node dissection (ELND) for intermediate-thickness melanoma (1-4 mm) improves survival rates compared with clinical observation of the lymph nodes. A second objective was to define subgroups of melanoma patients who would have a higher survival with ELND. METHODS: The eligible patients were stratified according to tumor thickness, anatomic site, and ulceration, and then were prerandomized to either ELND or nodal observation. Femoral, axillary, or modified neck dissections were performed using standardized surgical guidelines. RESULTS: The median follow-up was 7.4 years. A multifactorial (Cox regression) analysis showed that the following factors independently influenced survival: tumor ulceration, trunk site, tumor thickness, and patient age. Surgical treatment results were first compared based on randomized intent. Overall 5-year survival was not significantly different for patients who received ELND or nodal observation. However, the 552 patients 60 years of age or younger (75% of total group) with ELND has a significantly better 5-year survival. Among these patients, 5-year survival was better with ELND versus nodal observation for the 335 patients with tumors 1 to 2 mm thick, the 403 patients without tumor ulceration, and the 284 patients with tumors 1 to 2 mm thick and no ulceration. In contrast, patients older than 60 years of age who had ELND actually had a lower survival trend than those who had nodal observation. When survival rates were compared based on treatment actually received (i.e., including crossover patients), the patients with significantly improved 5-year survival rates after ELND included those with tumors 1 to 2 mm thick, those without tumor ulceration, and those 60 years of age or younger with tumors 1 to 2 mm thick or without ulceration. CONCLUSION: This is the first randomized study to prove the value of surgical treatment for clinically occult regional metastases. Patients 60 years or age or younger with intermediate-thickness melanomas, especially with nonulcerative melanoma and those with tumors 1 to 2 mm thick, may benefit from ELND. However, because some patients still are developing distant disease, these results should be considered an interim analysis.
Subject(s)
Lymph Node Excision/methods , Melanoma/surgery , Female , Follow-Up Studies , Humans , Lymphatic Metastasis , Male , Melanoma/mortality , Melanoma/secondary , Middle Aged , Prospective Studies , Regression Analysis , Survival RateABSTRACT
The role of surgery as primary treatment for patients with squamous cell carcinoma of the esophagus (SCCE) has been challenged by an improved response rate for radiotherapy that is made possible by adding radiosensitizing chemotherapy. The purpose of our study was to review our institution's treatment results for SCCE and to compare results of radiation versus surgery as primary treatment of early stage disease. A retrospective chart review was done on 241 patients who were treated with SCCE at Kansas University Medical Center and affiliated hospitals between 1970 and 1990. Patients were divided into five groups based on treatment received: (A) No Treatment; (B) Surgery Only; (C) Surgery plus Adjuvant Chemoradiotherapy; (D) Radiation Therapy Only; and (E) Chemoradiotherapy. Surgical treatment groups B and C had the best overall survival of all groups. To reduce any bias due to stage differences in groups, survival of groups was assessed only for early stage disease patients (Stage I, IIa, IIb). For Stage I and II patients receiving surgery as primary treatment (groups B and C), 29 per cent had a survival at 5 years compared to patients receiving primary radiation treatment (groups D and E) who had a combined survival of only four per cent. Although attempting comparison of risk groups is always a problem in nonrandomized studies, it is significant that only one 5-year survivor was in the nonresection radiation treatment groups D and E. Surgical resection for SCCE had the best survival in our study, especially in patients with early stage disease.
Subject(s)
Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/therapy , Esophageal Neoplasms/mortality , Esophageal Neoplasms/therapy , Adult , Aged , Aged, 80 and over , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Combined Modality Therapy , Humans , Middle Aged , Neoplasm Staging , Radiotherapy Dosage , Retrospective Studies , Survival Rate , Treatment OutcomeABSTRACT
PURPOSE: To determine whether the addition of surgical ovariectomy to standard chemotherapy prolongs disease-free survival (DFS) and overall survival in premenopausal patients with estrogen receptor (ER)-positive operable breast cancer with positive axillary nodes. PATIENTS AND METHODS: Three hundred fourteen premenopausal patients with ER-positive, node-positive breast cancer were enrolled between July 1979 and July 1989. Patients were stratified according to number of involved nodes and type of primary surgery and randomized to receive either of the following: (1) cyclophosphamide 60 mg/m2/d by mouth for 1 year, methotrexate 15 mg/m2 intravenously (i.v.) weekly for 1 year, fluorouracil (5-FU) 400 mg/m2 i.v. weekly for 1 year, vincristine .625 mg/m2 i.v. weekly for the first 10 weeks, and prednisone weeks 1 to 10 with doses decreasing from 30 mg/m2 to 2.5 mg/m2 (CMFVP); or (2) bilateral ovariectomy followed by CMFVP. RESULTS: The median follow-up time is 7.7 years and the maximum 13.2 years. Treatment arms are not significantly different with respect to either survival or DFS (one-sided log-rank, P = .55 and .70, respectively). The 7-year survival rate is 71% on the CMFVP arm and 73% on CMFVP plus ovariectomy. No significant differences were observed in node or receptor level subsets. CONCLUSION: We conclude that, in this study, the addition of ovariectomy did not improve results over chemotherapy alone in the treatment of premenopausal women with node-positive, ER-positive, operable breast cancer. Our sample size was too small to detect a small improvement. The death hazards ratio of CMFVP/CMFVP plus ovariectomy was 1.22 (95% confidence interval [CI], .79 to 1.89).