Biomedical subjects

J Cummings

Publications and source records attributed to J Cummings.

At least 91 records · Page 5Linked to original sources

PubMed1995

Effect of human recombinant interferon-alpha on the activity of cis-diamminedichloroplatinum(II) in human non-small cell lung cancer xenografts.

Interferons (IFNs) augment the effect of some antitumor agents, including cis-diamminedichloroplatinum(II) (cDDP), in experimental systems. The effect of human recombinant interferon-alpha 2b (rIFN alpha) on the cDDP-dependent growth delay of a human non-small cell lung cancer established as a xenograft in nude mice (NX002) has been investigated. IFN (10(5) IU/mouse, s.c.) as a single agent had no effect on the growth of the xenograft. cDDP (4.2 mg/kg, i.p.) caused a specific growth delay of 0.42, and this delay was significantly enhanced (to 1.08) by concomitant dosing with the otherwise inactive IFN. Possible mechanisms for this supra-additive relationship between IFN and cDDP have been investigated: increased intratumoral accumulation of platinum was seen at late time points (maximally at 36 hr) during the pharmacokinetic beta-phase of cDDP elimination from the plasma of the nude mice. Tumor:plasma platinum concentration ratios at 36-48 hr indicated significantly increased accumulation of platinum in tumors from IFN-treated mice compared to controls (p < 0.05). Scheduling experiments suggest that this IFN-mediated effect can persist for 4 hr. These differences may account for the enhanced antitumor activity of cDDP when coadministered with IFN.

Animals↗

PubMed1995

Identification of anthracenyl-dipeptide conjugates as novel topoisomerase I and II inhibitors and their evaluation as potential anticancer drugs.

As part of an ongoing rational drug design programme aiming to develop monosubstituted anthracenyl-peptides as potential anticancer drugs, three novel dipeptide conjugates have been synthesized and evaluated as inhibitors of topoisomerase (topo) I and II. Each of the three conjugates (designated NU/ICRF 600-602) was shown to inhibit the catalytic activity of both topoisomerase I and II, of which NU/ICRF 602 was the most active [100% inhibition of both enzymes at 5 micrograms/ml (approximately 15 microM) or less]. In a topo I/DNA unwinding assay, none of the compounds bound to DNA, suggesting genuine inhibition of catalytic activity. NU/ICRF 600 stabilized topo I cleavable complexes, although none of the compounds induced topo II-mediated DNA cleavage. Using a panel of Chinese hamster ovary cell lines along with the human ovarian cancer cell line, A2780, none of the three compounds were actively cytotoxic at concentrations < 100 microM. Subsequent drug uptake studies with NU/ICRF 600 and 602, using a method developed to correlate the chemosensitivity of A2780 cells with the uptake of anthracenyl-amino acid conjugates, revealed a lack of cellular uptake for both dipeptide conjugates. The significance of this finding in relation to drug design and the future development of this series of compounds is discussed.

Animals↗

PubMed1994

Sensitive determination of 8-chloroadenosine 3',5'-monophosphate and 8-chloroadenosine in plasma by high-performance liquid chromatography.

8-Chloroadenosine 3',5'-monophosphate (8-Cl-cAMP) is progressing through clinical evaluation as an anticancer drug. There is debate as to whether 8-Cl-cAMP is the active principal or its cytotoxic metabolite 8-Cl-adenosine. Separate high-performance liquid chromatographic methods are described for (i) 8-Cl-cAMP and its nucleotide metabolites (with 8-Br-cAMP as internal standard), and (ii) 8-Cl-adenosine. Both methods use a reversed-phase (Spherisorb ODS-2) stationary phase and a mobile phase consisting of sodium phosphate buffer (10 mM, pH 3.5) and methanol but with gradient elution for the nucleotides and isocratic elution for 8-Cl-adenosine. 8-Cl-cAMP and related nucleotides are extracted from plasma using strong anion-exchange solid-phase extraction (SPE) and 8-Cl-adenosine is extracted using reversed-phase (C8) SPE. Both techniques enabled analyses to be performed at high detector sensitivity with minimal interference. Limit of detection in plasma was 10 ng/ml for both 8-Cl-cAMP and 8-Cl-adenosine. When applied to the analysis of plasma samples from a patient treated with a low dose continuous infusion of 25 micrograms/kg/h, steady-state concentrations centred around 60 ng/ml 8-Cl-cAMP were determined. In the same patient 8-Cl-adenosine was not detected. Application of this methodology will aid in the further development of 8-Cl-cAMP as a potential new form of anticancer treatment.

2-Chloroadenosine↗

PubMed1994

Encapsulation of mitomycin C in albumin microspheres markedly alters pharmacokinetics, drug quinone reduction in tumour tissue and antitumour activity. Implications for the drugs' in vivo mechanism of action.

Pharmacokinetics and metabolism of mitomycin C (MMC) have been studied in NMRI mice bearing MAC 16 colon adenocarcinoma after direct intratumoural injection of either 500 micrograms free MMC or the same dose incorporated in albumin microspheres. Microspheres produced a tumour pharmacokinetic profile of steady state drug levels, avoiding the much higher early peak (20.5 micrograms/tumour vs 98.9 micrograms/tumour) and lower trough of free MMC, and reducing significantly the levels of drug reaching the systemic circulation (AUC 1.8 micrograms/mL x hr for microspheres vs 6.8 micrograms/mL x hr for free drug). 2,7-Diaminomitosene (2,7-DM), a key intermediate in MMC quinone bioreduction, was used as an indicator of drug metabolic activation in tumour tissue. Peak levels were 10-fold higher (11.2 micrograms/tumour vs 1.1 micrograms/tumour) and area under the curve 5-fold higher after free drug. Even taking into account differences in tumour pharmacokinetic profiles of the parent drug, microspheres actively inhibited 2,7-DM formation 3-fold. However, the microspheres generated a completely different pattern of drug metabolism where four previously uncharacterized mitosane metabolites and elevated levels of cis and trans 1-hydroxy 2,7-diaminomitosene were detected. Despite similar parent drug exposure in tumours, free drug was significantly more active (P < 0.05, Student's t-test) against MAC 16. These results suggest that formation of 2,7-DM correlates more closely with antitumour activity than sustained parent drug levels or appearance of other key metabolites. Potentially, they provide the first direct evidence for an in vivo mechanism of action dependent on bioreductive activation and formation of 2,7-DM.

Adenocarcinoma↗

PubMed1994

Changing subunit composition of heteromeric NMDA receptors during development of rat cortex.

Activation of the N-methyl-D-aspartate (NMDA) receptor is important for certain forms of activity-dependent synaptic plasticity, such as long-term potentiation (reviewed in ref. 1), and the patterning of connections during development of the visual system (reviewed in refs 2, 3). Several subunits of the NMDA receptor have been cloned: these are NMDAR1 (NR1), and NMDAR2A, 2B, 2C and 2D (NR2A-D). Based on heterologous co-expression studies, it is inferred that NR1 encodes an essential subunit of NMDA receptors and that functional diversity of NMDA receptors in vivo is effected by differential incorporation of subunits NR2A-NR2D. Little is known, however, about the actual subunit composition or heterogeneity of NMDA receptors in the brain. By co-immunoprecipitation with subunit-specific antibodies, we present here direct evidence that NMDA receptors exist in rat neocortex as heteromeric complexes of considerable heterogeneity, some containing both NR2A and NR2B subunits. A progressive alteration in subunit composition seen postnatally could contribute to NMDA-receptor variation and changing synaptic plasticity during cortical development.

Animals↗

PubMed1994

Determination of two neuropeptide growth factor antagonists, [D-Arg1,D-Phe5,D-Trp7,9,Leu11]-substance P and [Arg6,D-Trp7,9,N-MePhe8]- substance P(6-11), by high-performance liquid chromatography with electrochemical detection.

The neuropeptide growth factor antagonists [D-Arg1,D-Phe5,D-Trp7,9,Leu11]-substance P (D) and [Arg6,D-Trp7,9, [corrected] N-MePhe8]-substance P(6-11) (G) are currently undergoing preclinical evaluation as potential anticancer agents and clinical trials are planned for G in the near future. A reversed-phase high-performance liquid chromatographic separation has been developed which is both sensitive (limit of detection 250 pg/263 fmol for G; 500 pg/330 fmol for D) and selective, based on electrochemical detection of the two tryptophan residues present in each peptide. Two ion-pairing agents were included in the isocratic mobile phase to eliminate adsorption of the peptides onto the analytical column. Extensive sample clean-up procedures have been developed for plasma, tissue and tumour based on solid-phase extraction. Precision and accuracy of each assay was 91.3 +/- 16.9% (between-day) for G and 99.3 +/- 16.9% (between-day) for D. The assays were able to detect the intact peptides and a number of their metabolites in plasma, liver and the WX 322 SCLC human xenograft in nude mice for at least 6 hr after administration of therapeutic and pharmacological doses.

Amino Acid Sequence↗

PubMed1994

Stability and in vitro metabolism of the mitogenic neuropeptide antagonists [D-Arg1,D-Phe5, D-Trp7,9, Leu11]-substance P and [Arg6, D-Trp7,9, MePhe8]-substance P (6-11) characterized by high-performance liquid chromatography.

The substance P (SP) analogues [D-Arg1, D-Phe5, D-Trp7,9, Leu11]-SP and [Arg6, D-Trp7,9, MePhe8]-SP (6-11) (antagonists D and G, respectively) are under consideration as new anticancer drugs. In this report, the stability and in vitro metabolism of both antagonists in up to seven different media (water, 1 M acetic acid, human plasma, nude mouse liver and WX 322 human SCLC xenograft homogenized in either 1 M acetic acid or phosphate buffered saline (PBS), pH 7.4) have been characterized by both isocratic and gradient elution reversed-phase HPLC. Antagonist D was stable (never > 13% degradation over 24 h, at 37 degrees C) in water, 1 M acetic acid and plasma but was metabolized by PBS liver homogenates (10%, w/v) sequentially to two stable metabolites with a half life of 0.98 h at a concentration of 500 micrograms ml-1. The major pathway of degradation of antagonist G appeared to be C-terminal methionine oxidation (particularly in plasma) as well as hydrolysis, with even aqueous solutions being significantly affected at low concentrations of peptide (0.1 micrograms ml-1, half life 20.9 h at 37 degrees C). Stable metabolites of antagonist G were also detected in incubations with PBS liver homogenates (half life 1.53 h at 500 micrograms ml-1, 37 degrees C). Overall, the data presented indicate that the modifications made to SP have been relatively successful in preserving chemical and biological stability.

Amino Acid Sequence↗

PubMed1994

Pharmaceutical analysis of 8-chloroadenosine 3',5'-monophosphate.

8-Chloroadenosine 3',5'-monophosphate (8-Cl-cAMP) has recently entered clinical trials as a new anticancer drug. 8-Cl-cAMP can undergo metabolism/degradation to nucleotides (8-Cl-AMP; 8-Cl-IMP), nucleosides (8-Cl-adenosine; 8-Cl-inosine) and bases (8-Cl-adenine; 8-Cl-xanthine and 8-Cl-hypoxanthine). A reversed-phase, gradient elution high-performance liquid chromatographic method is described which can resolve all the above from their non-chlorinated naturally occurring counterparts and 8-Bromo-cAMP. The assay was then utilized to perform pharmaceutical evaluations on the 8-Cl-cAMP formulation. Sterilization by filtration (0.22 micron pore size) resulted in no loss of material. Stability studies carried out in the cartridges used to continuously infuse (for 7 days) the drug to patients showed no significant degradation over 17 days. Purity determinations revealed the presence of up to nine impurities (related to adenine) and yielded a purity figure of 95.9-99.3% with considerable batch to batch variation.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1994

Subcortical disease and neuropsychiatric illness.

The relationship between neuropsychiatric disorders and dysfunction of subcortical structures is the theme of the 1994 joint meeting of the British and American Neuropsychiatric Associations, to be held July 21-24, 1994, in Newport, Rhode Island. In this essay Dr. Salloway, chair of the Scientific Program Committee, and Dr. Cummings, director-elect of The American Neuropsychiatric Association, define the structures embraced in the "subcortical" concept and summarize the relationship between specific neuropsychiatric symptoms and regional subcortical dysfunction. These concepts and observations will be expanded, challenged, and refined at the meeting. (See the Preliminary Annual Meeting Program in this issue.)

Brain↗

PubMed1994

Customized dual data entry for computerized data analysis.

A major responsibility of any Quality Assurance Unit (QUA) is to ensure data integrity. Errors made during data entry can lead to many problems in the study review process and decrease the quality, accuracy, and overall efficiency of data management. One technique that can reduce the number of data entry errors in computer data sets is the use of a dual entry data system. Currently available software allows creation of customized data entry screens that either closely resemble or duplicate the data collection forms used during studies. Two data entry operators enter data into two independent data sets. The use of an on-screen display that resembles the data collection form reduces the potential for keypunch errors. The two data sets can then be electronically compared. The comparison reports differences between the two data sets. When differences exist, the correct values can be determined by reference to the original data sheets and the two data files can then be corrected. Theoretically, the only key punch errors that will exist after making these corrections are when the two independent entry operators make the same exact data entry error. Typically, the time required for two people to enter data is minimal compared to the time required to manually identify and correct data entry discrepancies. With error-free data entry, we have found that electronic data quality, accuracy, and audit efficiency are improved at every subsequent step of data management, analysis, quality assurance auditing, and report generation.

Information Systems↗

PubMed1993

Application of high-performance liquid chromatography for recognition of covalent nucleic acid modification with anticancer drugs.

Covalent modification of DNA by antineoplastic agents represents a potent biochemical lesion which can play a major role in drug mechanism of action. The ability to measure levels of DNA covalent modifications in target cells in vivo may, therefore, be seen as the ultimate form of therapeutic drug monitoring. Additionally, elucidation of the structure of critical DNA adducts and definition of their role in tumour cell cytotoxicity will provide more selective targets for rational drug design of new cancer chemotherapeutic agents. High-performance liquid chromatography has contributed significantly to all these areas. In vivo levels of nucleic acid covalent modifications are in the range of 1 in 10(5)-10(8) nucleotides precluding the use of conventional high-performance liquid chromatographic detection methods. Several classes of natural product anticancer drugs have been shown to bond covalently to nucleic acids under optimal laboratory conditions. These have proved more accessible to high-performance liquid chromatographic analysis because of their lipophilicity and strong UV chromophores. However, the majority of experimental evidence to date suggests that with the exception of mitomycin C and morpholino-anthracyclines these compounds do not exert their primary mechanism of action through nucleic acid covalent modification. DNA adducts of alkylating and platinating agents are more difficult to detect by high-performance liquid chromatography and can be chemically unstable. These compounds interact with DNA on the basis of chemical kinetics. Thus, the principle sites of attachment tend to be with the most nucleophilic base (guanine) at its most reactive centre (N-7 position). Limited in vivo high-performance liquid chromatographic studies with all classes of anticancer drugs indicate a much more complex pattern of adductation than would have been anticipated from in vitro studies alone. Some of these differences are probably due to methodological artefacts but these studies stress the need for sensitive detection methods and reliable sample preparation (nucleic acid extraction and digestion techniques) when attempting to determine nucleic acid covalent modifications by anticancer drugs.

Animals↗

PubMed1993

Examination gloves as barriers to hand contamination in clinical practice.

OBJECTIVE: To test the effectiveness of vinyl and latex gloves as barriers to hand contamination with gram-negative organisms and enterococci during routine hospital procedures. DESIGN AND INTERVENTIONS: We studied 137 procedures during which a health care worker's gloved hand contacted a patient's mucous membrane and was thus potentially contaminated with gram-negative rods or enterococci. Quantitative hand cultures were obtained from each health care worker before and after the gloved contact using a modified glove juice method, and the exterior glove surface was also quantitatively cultured after patient contact. Used gloves were then tested for leaks using the American Society for Testing and Materials' watertight test. SETTING: Harborview Medical Center, a 330-bed city-county hospital and level I regional trauma and burn center, is both a teaching facility affiliated with the University of Washington and the major provider of care to indigent and uninsured persons in Seattle-King County, Washington. PATIENTS AND OTHER PARTICIPANTS: Respiratory therapists performing endotracheal tube care on intubated intensive care unit patients, registered nurses performing digital rectal stimulation for bowel training on patients with spinal cord injury in the rehabilitation ward, and dentists performing routine dental examinations and procedures on healthy outpatients in the dental clinic. MAIN OUTCOME MEASURE AND RESULTS: Eighty-six of the 135 gloves cultured had gram-negative rods or enterococci on the external surface after use and were thus sources of potential hand contamination. Microbial contamination of the health care worker's hands occurred in 11 (13%; 95% confidence interval, 6% to 20%) of these 86 events, and was more frequent with vinyl (10 of 42) than latex (one of 44) gloves (P < .01). After use, glove leaks were also more frequent in vinyl gloves (26 of 61) than with latex gloves (six of 70) (P < .001). Even when leaks were present, gloves prevented hand contamination in 77% of instances and quantitative counts of microorganisms contaminating hands were 2 to 4 logs less than counts on external glove surfaces. Health care workers reported awareness of the presence of glove leaks in only seven (22%) of the 32 events in which leaks were subsequently demonstrated. CONCLUSIONS: Under conditions of routine use, gloves effectively function as a protective barrier even when leaks are present. Latex gloves were less frequently associated with leaks and hand contamination. Since hand contamination occurred after 13% of exposures and cannot be readily identified by health care workers, routine hand washing should be done after each patient contact.

Enterococcus↗

PubMed1993

Correlation between tumour drug disposition and the antitumour activity of doxorubicin-loaded microspheres: implications for the drugs' in vivo mechanism of action.

Doxorubicin (DOX) has been incorporated into five different formulations of protein microspheres, each altering tumour drug disposition in a characteristic manner. There was no correlation between the stimulation in anaerobic quinone bioreduction over the levels produced by free DOX and tumour growth delay against the Sp 107 rat mammary carcinoma. A strong correlation (r2 = 0.948, P < 0.01, two-tailed t statistic) was observed between a slower decline in parent drug levels and antitumour activity. These data support the view that free radical processes are not involved in the mechanism of action of DOX and suggest that the optimum way to delivery the drug to the Sp 107 tumour is through sustained release of lower concentrations (approximately 2 microM) from a large extracellular pool.

Animals↗

PubMed1993

Determination of mitomycin C, 2,7-diaminomitosene, 1,2-cis- and 1,2-trans-1-hydroxy-2,7-diaminomitosene in tumour tissue by high-performance liquid chromatography.

A high-performance liquid chromatographic method is described for the determination of mitomycin C (MMC) and its metabolites 2,7-diaminomitosene (2,7-DM), 1,2-cis-1-hydroxy-2,7-diaminomitosene (cis-hydro) and 1,2-trans-1-hydroxy-2,7-diaminomitosene (trans-hydro) in tumour tissue. N-la-Methylmitomycin C (porfiromycin, PM) was used as an internal standard. Two factors were critical in resolving the metabolites: pH and buffer ionic strength, where the retention times of the four components were affected in the order 2,7-DM >> cis-hydro >> trans-hydro >> MMC. The optimal isocratic conditions (flow-rate 1 ml/min) were 18 mM sodium phosphate pH 5.8-methanol (74:26) and a column temperature of 40 degrees C on a Spherisorb ODS-2 column (25 cm x 4.6 mm I.D.). Liquid-liquid extraction [twice with chloroform-propan-2-ol-ethyl acetate (2:2:1)] is described for tumour tissue. Recoveries varied depending on the component: MMC, 71.9 +/- 12.4%; PM, 85.5 +/- 27%; 2,7-DM, 51.7 +/- 5.4%; cis-hydro, 52.0 +/- 16.8%; trans-hydro, 62 +/- 8%. When applied to the analysis of a rat mammary carcinoma treated intra-tumourally with 450 micrograms of MMC five drug-related "metabolite" peaks were detected. Three of these co-chromatographed with standards of 2,7-DM, cis- and trans-hydro, and had identical absorption maxima to their respective standards, with the possible exception of trans-hydro.

Animals↗

PubMed1993

The comparative disposition of the pyrolloquinone GR63178A and its 9-hydroxy metabolite GR54374X in sensitive and resistant mouse colon adenocarcinoma.

The novel anticancer compound GR63178A is being evaluated in the clinic, having demonstrated activity against a wide range of experimental tumour systems in animals without significant toxic side-effects being apparent. In this work, we have demonstrated significant antitumour action of this compound against one murine colon cancer model (colon 38 tumour in BDF-1 mice, specific growth delay = 1.2) when given at 10 mg/kg over 21 consecutive days and in contrast shown minimal sensitivity of another similar murine colon adenocarcinoma, MAC 26, in NMRI mice with the same dose regime. We investigated the disposition of both the parent drug and the 9-OH metabolite (GR54374X) in plasma, tissues and tumours, using solid phase extraction followed by reversed-phase high performance liquid chromatography. Although plasma clearance profiles of GR63178A were similar, significant differences were seen in the disposition of the drug to major organs in two mouse strains. Noteably, the liver and kidneys of the sensitive model had higher levels of parent drug and 9-OH metabolite at both 30 min and 4 h post-injection. However, this was not apparent in the tumours themselves, and the levels of 9-OH metabolite were lower in the plasma and higher in the urine of the sensitive mice, indicating possible rapid renal clearance of this compound. Neither GR63178A nor GR54374X proved cytotoxic in in vitro experiments. The data presented here have revealed considerable variation in drug handling by these two mouse strains, but this did not produce different levels of either parent drug or GR54374X in the tumours, which are the presumed targets, suggesting that differences in disposition are probably not responsible for the different sensitivities of the two tumours. Other possible explanations include the production of a hitherto undetected ultimate cytotoxic metabolite in the sensitive, but not in the resistant, mouse/tumour combination, or differences in inherent tumour sensitivity, or in host-mediated effects. These possibilities are discussed.

Adenocarcinoma↗

PubMed1993

Endoluminal urethral stents: a review.

Although Fabian first introduced the concept of an endourethral stent in 1980, recent developments in biomedical technology and the treatment philosophy of urethral obstruction has led to a resurgence of this concept. We review the past and current literature with regard to both temporary and permanent stents. Available stents are described, clinical results summarized, and indications discussed.

Equipment Design↗

PubMed1993

DNA topoisomerase I and II as targets for rational design of new anticancer drugs.

BACKGROUND: Topoisomerase I and II (topo I and II) are enzymes which alter the topological state of DNA through DNA strand cleavage, strand passage and religation. They participate in most aspects of DNA metabolism and are therefore vital to the cell undergoing division. Only one form of topo I has been identified whereas two isoenzymes of topo II have been described: the alpha form (170 kDa protein) and beta form (180 kDa protein). Both topo II isoenzymes have distinct nuclear localisation, are regulated independently, differ in their responsiveness to inhibitors and are differentially expressed in drug resistant cell lines. RESULTS: Several clinically active anticancer drugs (e.g., doxorubicin, m-AMSA, VP-16 and camptothecins) poison these enzymes by stabilizing a putative reaction intermediate called the cleavable complex (cc) where the topoisomerase remains covalently attached to either one strand of DNA (topo I) or both strands of double helix (topo II) after strand cleavage. DNA cleavage sites appear unique for different classes of inhibitor, and are probably critical for defining cytotoxicity. Formation of the cc may cause cell death either by colliding with replication forks, by promoting illegitimate genomic-DNA recombination, by arresting cells in the G2-phase of the cell cycle or by inducing apoptosis. CONCLUSION: New classes of inhibitor have recently been described with novel mechanisms of action including compounds which do not stabilize cleavable complexes or bind significantly to DNA. These may prove to be more selective and less toxic. They may also avoid the possible problem of therapy-related leukemias associated with topo inhibitors which induce DNA cleavage and chromosomal aberrations.

Animals↗

PubMed1993

The long term stability of mechlorethamine hydrochloride (nitrogen mustard) ointment measured by HPLC.

HPLC has been applied to determine the stability of mechlorethamine hydrochloride (nitrogen mustard) formulated as an ointment in white soft paraffin (10 mg drug, 50 g paraffin and 1 mL acetone). A new solubilization technique is described for extraction of the drug from the ointment for HPLC analysis which has an extraction efficiency of 76.1% with a coefficient of variation of 10.4%. Stored at 4 degrees C the drug content of the ointment remained stable for at least 84 days and stored at 37 degrees C the drug content remained stable for at least 40 days. In comparison aqueous solutions of mechlorethamine at the same concentration, fully degraded after 4 days. These data will aid hospital pharmacists decide on a standardized protocol for the controlled usage of the ointment which is both safe and cost-effective.

Chromatography, High Pressure Liquid↗