Cytotoxic effects of topotecan combined with various anticancer agents in human cancer cell lines

BACKGROUND: Topotecan (TPT) is a topoisomerase I poison that exhibits antineoplastic activity. Analysis of the cytotoxic effects of combinations of TPT and other anticancer agents has been limited. PURPOSE: We assessed the cytotoxic effects produced by combinations of TPT and other antineoplastic agents in experiments involving multiple human cancer cell lines of diverse histologic origins. METHODS: The cytotoxic effects of various antimetabolites (fluorouracil, methotrexate, or cytarabine), antimicrotubule agents (vincristine or paclitaxel [Taxol]), DNA alkylating agents (melphalan, bis[chloroethyl]nitrosourea [BCNU], or 4-hydroperoxycyclophosphamide [4HC]), and a DNA-platinating agent (cisplatin), alone and in combination with TPT, were measured in clonogenic (i.e., colony-forming) assays. HCT8 ileocecal adenocarcinoma, A549 non-small-cell lung carcinoma, NCI-H82ras(H) lung cancer, T98G glioblastoma, and MCF-7 breast cancer cell lines were used in these assays. The data were analyzed by the median effect method, primarily under the assumption that drug mechanisms of action were mutually nonexclusive (i.e., completely independent of one another). For each level of cytotoxicity (ranging from 5% to 95%), a drug combination index (CI) was calculated. A CI less than 1 indicated synergy (i.e., the effect of the combination was greater than that expected from the additive effects of the component agents), a CI equal to 1 indicated additivity, and a CI greater than 1 indicated antagonism (the effect of the combination was less than that expected from the additive effects of the component agents). RESULTS: When the mechanisms of drug action were assumed to be mutually nonexclusive, virtually all CIs for combinations of TPT and either antimetabolites or antimicrotubule agents revealed cytotoxic effects that were less than additive. The CIs calculated at low-to-intermediate levels of cytotoxicity for combinations of TPT and the DNA alkylating agents melphalan, BCNU, and 4HC also showed drug effects that were less than additive; in most cases, however, nearly additive or even synergistic effects were observed with these same drug combinations at high levels of cytotoxicity (i.e., at > or = 90% inhibition of colony formation). Results obtained with combinations of TPT and cisplatin varied according to the cell line examined. With A549 cells, less than additive effects were seen at low-to-intermediate levels of cytotoxicity, and more than additive effects were seen at high levels of cytotoxicity. With NCI-H82ras(H) cells, synergy was observed over most of the cytotoxicity range. CONCLUSIONS AND IMPLICATIONS: TPT cytotoxicity appears to be enhanced more by combination with certain DNA-damaging agents than by combination with antimetabolites or antimicrotubule agents. Interactions between TPT and other drugs can vary depending on the cell type examined. Further investigation is required to determine the basis of the observed effects and to determine whether these in vitro findings are predictive of results obtained in vivo.     

CT of appendicitis in children

PURPOSE: Our goal was to review the CT findings and to help define the role of CT in the evaluation of appendicitis in children. METHOD: Of 730 children with surgically proven appendicitis, 22 underwent preoperative CT evaluation. Their CT scans and operative and pathology records were retrospectively reviewed. The CT scans were evaluated for appendiceal wall thickness, diameter, and location, appendicoliths, pericecal inflammation, phlegmon, abscess, free fluid, small bowel dilatation, and bowel wall thickening. Criteria for diagnosing appendicitis were (a) appendiceal wall thickening (> 1 mm) or (b) presence of abscess, phlegmon, or pericecal inflammation associated with appendicolith(s). Prospective reports of ultrasound examinations performed within 2 days of the CT scans were available in 14 children and were correlated with the CT findings. RESULTS: An abnormally thickened appendix, with a diameter ranging from 9 to 18 mm, was seen in four children. Three appendices were retrocecal and one was near the cecal tip, anterior to the iliac vessels. Appendicoliths were present in 10 children, multiple in 1. Abscesses were seen in 13 of 22 children, multiple in 5. Phlegmon was seen in five children and pericecal inflammation in two. Bowel wall thickening was present in seven children and small bowel dilatation was noted in six. Other findings included free fluid, hydronephrosis, thickening of urinary bladder wall, air in the uterus and vagina, adenopathy, and thickening of the abdominal wall musculature. CT was diagnostic of appendicitis in 11 of 22 children (50%). In 14 children with both ultrasound and CT studies, CT was slightly better in diagnosing appendicitis and visualizing the abnormal appendix and was superior in defining the presence and extent of abscess and inflammation in 9 of 14 children. CONCLUSION: CT is a useful adjunct in diagnosing appendicitis in children, with a major role in cases of complicated appendicitis.     

Plasmid DNA is protected against ultrasonic cavitation-induced damage when complexed to cationic liposomes

Cationic liposomes bound to plasmid DNA are currently used for in vitro and in vivo gene therapy applications, but such complexes readily form large, heterogeneous aggregates that are not appropriate for pharmaceutical development. More importantly, size heterogeneity makes studies focused on optimizing gene transfer to cells difficult to conduct or understand. For this reason we have evaluated the effect of microprobe sonication on these complexes in an effort to achieve process-controlled size homogeneity. Complexes were prepared using a 7.2 kb reporter plasmid and the following liposomal lipid combinations: DDAB/DOPE (50:50 mol %), DDAB/DOPE/PEG-PE (50:45:5 mol %), DDAB/EPC (50:50 mol %), DDAB/EPC/PEG-PE (50:45:5, 50:40:10, 50:35:15 mol %), DODAC/DOPE (50:50 mol %), and DODAC/EPC (50:50 mol %) (DDAB, dimethyldioctadecylammonium bromide; DOPE, dioleoylphosphatidylethanolamine; PEG-PE, monomethoxypolyethylene glycol2000 succinate- distearoylphosphatidylethanolamine; EPC, egg phosphatidylcholine; DODAC, dioleoyldimethylammonium chloride). The influence of complex composition and lipid:DNA ratio was evaluated. Particle size was determined before and after complexation and again after sonication using the quasi-elastic light scattering technique. DNA integrity was assessed via agarose gel electrophoresis. Finally, gene transfection was evaluated using CHO cells that were transfected in vitro with sonicated and unsonicated complexes. It is established in this study that size reduction can occur, but this is dependent on cationic and neutral lipid composition and, in some cases, lipid:DNA ratio. Surprisingly, the process of sonication leaves a significant percentage of the plasmid DNA intact and capable of in vitro transfection. This study shows that plasmid DNA can be protected from damage due to sonication by liposome complex formation. This may indicate that more common pharmaceutical methods for size reduction which subject particles to mechanical stress may be applicable in preparation of liposome/DNA formulations for in vivo application.     

Improved accuracy of burn wound assessment using laser Doppler

The utility of the laser Doppler for determining burn depth has been questioned because of problems with technology and methodology. This study prospectively evaluates the ability of a new laser Doppler technique to predict burn healing time. Using the Periflux System 4000 laser Doppler, readings were taken on 305 burns (147 patients) on postburn day 3 or 4. Sixty-six wounds were used to derive a predictive function (phase I) and 152 wounds were used to test the function (phase II). Blood flow dynamics (flux), microvascular dilation capacity of the wounds to beat stress, and flow motion wave pattern (vasomotion) were studied using the laser Doppler, and seven parameters were evaluated to determine their relative contribution to the prediction of healing time. These parameters are hyperemic flux (flux value after heating to 42 degrees C), average hyperemic wave amplitude (AHWA), number of average flux units >100(F100), number readings with wave amplitude 75 (A5), average flux change (AFC), percentage of average flux increase, and relative flow capacity (RFC = AFC/average hyperemic flux). After readings were made, the wounds were observed and divided into two groups: those that healed in less than 14 days and those that healed or were grafted after 14 days. A step-wise discriminant analysis was used to assess the relative contribution of the Doppler-derived measures to healing time prediction. AHWA, F100, and RFC were included in the final discriminant function explaining 72% of the healing time variance (Wilks' lambda value 0.28; p value <0.0001). Predicted outcome = 0.05(AHWA) + 0.31(F100) + 5.0(RFC) - 2.3. With this derived function, there is 94% accuracy in the prediction of burn wound healing time compared with a physician predictive accuracy of 70%.     

Discrepancies between thyrotropin (TSH) measurement by four sensitive immunometric assays

A comparison of four different commercial immunometric thyrotropin (TSH) assays (Amerlite R TSH-30 Ultrasensitive assay from Kodak, BeriLux R hTSH from Behring Werke, Delfia R hTSH Ultra from Wallac and IMX R Ultrasensitive hTSH from Abbott) was made by measuring serum TSH in 81 consecutive patients referred to hospital for various reasons with a serum TSH less than 0.8 mlU/l in the IMX assay. The analytical and functional assay sensitivities of each of the assays were analysed. Even though three of the methods had a sensitivity corresponding to third generation assays, we could only demonstrate an overall agreement of serum TSH when comparing two of the kits. The measurements in Delfia Ultra and Berilux showed good agreement (P = 0.7, paired t-test and bias = 0.003 mIU/l), while the comparisons between the other assays showed different measurements (P < 0.00001, paired t-test and bias more than 0.07 mIU/l). Differences in the calibrators used in the assays might explain some of the discrepancy, although all methods were calibrated according to the same international standard. Also, differences in the specificity of the TSH monoclonal antibodies used in the assays might be an evident explanation and further studies of the specificity of the monoclonal antibodies are needed. An international collaborative study to clarify reasons for the differences between the TSH assays and to standardize the measurements is recommended.     

A general and flexible method of problem definition in structural optimization systems

This paper describes the implementation of a general and flexible method of formulating problems of mathematical programming in structural optimization systems. The method enables the formulation and solution of problems involving scalar, integral, min/max, max/min and possibly non-differentiable user defined functions in any conceivable mix. The mathematical formulation is based on the bound formulation, and the implementation specific details involve a parser capable of interpreting and performing symbolic differentiation of the user defined functions.     

Fertilization in vitro. A review over the medical indications and suggestion to unified guidelines for public fertility clinics in Denmark

Medical indications for in vitro fertilization and embryo transfer (IVF-ET) internationally and in Denmark are reviewed. Reports from large international centres document that tubal infertility, unexplained infertility, endometriosis and male infertility are equally good indications for IVF. Traditionally, tubal infertility has been the only medical indication qualifying for IVF treatment within the National Health Service in Denmark. Thus, in this country, couples with unexplained and male infertility and with endometriosis have to pay up to 25,000 D.Kr. per IVF-ET treatment in private fertility clinics. Since there is no scientific basis for this discrimination, it is urged that the present rules are changed, so that couples with unexplained and male infertility and endometriosis are also allowed IVF treatment free of charge in the public fertility clinics.     

Study of inaccuracy in semi-analytical sensitivity analysis — a model problem

The semi-analytical method of sensitivity analysis (Zienkiewicz and Campbell 1973; Esping 1983; Cheng and Liu 1987) of finite element discretized structures is attractive due to the balance between computational cost and ease of implementation (Cheng and Liu 1987; Haftka and Adelman 1989), but unfortunately the method may exhibit serious inaccuracies when applied in shape optimization of structures modelled by beam, plate, shell and Hermite elements (Cheng and Liu 1987; Haftka and Adelman 1989; Barthelemyet al. 1988; Barthelemy and Haftka 1988; Choi and Twu 1991, Pedersenet al. 1989; Chenget al. 1989).In the present paper, we perform an exact analysis of the error of sensitivity for a simple model problem which has earlier been considered by Barthelemyet al. (1988), Barthelemy and Haftka (1988), Pedersenet al. (1989). The analysis gives a deep insight into the nature of the general inaccuracy problem and enables us to devise methods by which the severe error of the sensitivity can be substantially reduced or removed for the model problem. The results of the paper are illustrated via an example.A method of error elimination for an extended class of semianalytical analysis problems is developed and presented in a companion paper (Olhoff and Rasmussen 1991).