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Biomedical subjects

J D Allen

Publications and source records attributed to J D Allen.

At least 19 recordsLinked to original sources
PubMed2001

Inhibition of BCRP-mediated drug efflux by fumitremorgin-type indolyl diketopiperazines.

A library of 42 diastereoisomeric mixtures of fumitremorgin-type indolyl diketopiperazines, prepared by parallel solid-phase synthesis, was screened for Breast Cancer Resistance Protein inhibitory activity and compared with GF120918. Demethoxy-fumitremorgin C was synthesized by solid-phase techniques and tested as well. Structure-activity relationship studies have identified several potent analogues, both in assays using the T6400 mouse and the T8 human cell line, whereas low cytotoxicity was seen at effective concentrations.

ATP Binding Cassette Transporter, Subfamily G, Mem↗
PubMed2001

Discrimination of gamma-hydroxybutyrate and ethanol administered separately and as a mixture in rats.

The physiological effects of gamma-hydroxybutyrate (GHB) are complex and not yet clearly defined. GHB has been labeled as a recreational drug and is reported to be frequently coabused with ethanol (ETH). Other studies have yielded discrepant results as to the interaction between GHB and ETH. Thus, the present study investigated extensively the discriminative stimulus of GHB and ETH and a mixture of the two compounds. Thirty male Long-Evans rats were divided into three groups and trained to discriminate doses of either 300 mg/kg GHB, 1000 mg/kg ETH, or a mixture (MIX: 150 mg/kg GHB+500 mg/kg ETH) from vehicle on a two-lever fixed-ratio (FR) 10 schedule of food reinforcement. Dose-response curves were attained in each group with its respective training drugs. GHB and ETH did not cross-generalize in the ETH- and GHB-trained rats, respectively. However, when the effects of the MIX were tested in the GHB- and ETH-trained rats, a greater than additive response was observed. Testing also revealed that the MIX-trained rats did not perceive a novel stimulus but a near-equal contribution from GHB and ETH. This study provides evidence of a complex relationship between GHB and ETH and opposes previous work reporting cross-generalization between GHB and ETH.

Adjuvants, Anesthesia↗
PubMed2001

Waveform optimization for internal atrial defibrillation: effects of waveform rounding, phase duration, and voltage swing.

The aim of this study was to compare the efficacy of internal atrial defibrillation by conventional truncated exponential biphasic waveforms with and without waveform rounding (1-2 phases) and to determine optimal duration for this novel double rounded waveform. Atrial fibrillation, induced by rapid electrical stimulation, was converted by internal shocks through defibrillation catheters (lateral right atrium and coronary sinus) in anesthetised sheep. Rounding the leading edges of the conventional biphasic waveform (Ventritex HVS-02; settings 100/-50 V, 150/-70 V, and 200/-100 V; n = 8) reduced delivered peak and trough voltages, currents, and energy (by > or = 21 %, P < 0.001; for double (both phases) rounded) without decreasing cardioversion success. At 100/-50 V the efficacy of single (first phase) rounded (53 +/- 13%; mean +/- SEM) and double rounded (59 +/- 11%) shocks was similar to the conventional waveform (56 +/- 14%). Double rounded waveform (phase durations 1-20 ms) efficacy was optimum at 6-10 ms phase duration (100% success at 10-ms phase duration; 1.52 +/- 0.04 J delivered energy; n = 6). Successful cardioversion by conventional, single rounded, and double rounded biphasic waveforms (duration 6 ms each phase), conventional monophasic, rounded monophasic (duration 12 ms), and a damped sine waveform correlated strongly with peak-to-trough voltage swing within the waveform (r = 0.882; P < 0.01; n = 8). For internal atrial defibrillation, rounding both phases of the conventional biphasic waveforms, the double rounded waveform, permits similar efficacy to the conventional truncated exponential biphasic waveform at reduced peak voltage, current, and delivered energy. Optimum phase duration is 6-10 ms (tested range 1-20 ms).

Animals↗
PubMed2001

Laparoscopic dismembered pyeloplasty: 50 consecutive cases.

OBJECTIVE: To test the hypothesis that laparoscopic dismembered pyeloplasty offers the same good results as open pyeloplasty, but without the disadvantages of the loin incision (which is painful, prolongs hospitalization and prevents a return to normal activities for several weeks) in the treatment of pelvi-ureteric junction (PUJ) obstruction. PATIENTS AND METHODS: Fifty consecutive consenting patients presenting with PUJ obstruction underwent laparoscopic dismembered pyeloplasty carried out by one surgeon using an extraperitoneal approach. RESULTS: Two (4%) procedures were converted to open surgery. The mean (range) operative duration was 164 (120-240) min. Fifteen (30%) of the patients had their ureter transposed anterior to a crossing lower-pole vessel; 22 (44%) patients had a separate renal pelvic suture line. The mean (range) postoperative parenteral analgesic requirement was 19.1 (0-111) mg of morphine sulphate. The mean (range) hospitalization was 2.6 (2-7) days. Two (4%) patients had complications. After a mean (range) follow-up of 18.8 (3-72) months all but one patient, who had failed endopyelotomy, had a normal renogram and were symptom-free. CONCLUSION: These results suggest that a loin wound is not necessary for a successful outcome after dismembered pyeloplasty, and that in expert hands laparoscopic dismembered pyeloplasty should now be considered the standard of care.

Adolescent↗
PubMed2001

Myocardial expression of the endothelin system in endotoxin-treated rats.

Although circulating plasma levels of endothelin (ET)-1 are elevated in endotoxemia, little is known about the myocardial expression of the ET system in endotoxic shock. We assessed the temporal mRNA expression pattern of key components of the ET system (pre-pro ET (ppET) -1, -2, ET-converting enzyme-1, ET(A) and ET(B) receptors) by reverse transcription polymerase chain reaction in a rat model of early endotoxic shock. Lipopolysaccharide (5 mg/kg, i.p.) caused a transient increase (p < 0.05) in inducible nitric oxide synthase mRNA expression. ppET-1 mRNA expression was increased at 2 h (approximately 12-fold increase; p < 0.05) in the lipopolysaccharide compared with the saline group and ppET-2 mRNA expression was unaltered. ET-converting enzyme-1, ET(A), and ET(B) receptor mRNA expression was unaltered in the lipopolysaccharide compared with the saline group. While ppET-1 mRNA expression is selectively upregulated in ventricular myocardium of lipopolysaccharide-treated rats, an absence of alteration in ET-converting enzyme-1 mRNA expression suggests an excess capacity of ET-converting enzyme-1 to cope with the increased expression of ET-1. At the level of the receptor, endotoxic shock did not affect the expression of either ET(A) or ET(B) receptor mRNA. These data are consistent with the increased expression of myocardial ET-1 as an acute-phase response due to hemodynamic instability associated with the early stages of endotoxic shock.

Animals↗
PubMed2001

Forearm vasoreactivity in type 1diabetic subjects.

This study compared forearm vasoreactivity in 15 Type I diabetic subjects with 15 healthy controls. The groups were matched for age, exercise capacity, and the absence of other cardiovascular risk factors. Vasoreactivity was measured using strain gauge plethysmography, at rest, after arterial occlusion (OCC), and following OCC coupled with handgrip exercise (ROCC). Forearm blood flows were significantly elevated between conditions 2.58 +/- 0.37ml/100mltissue at rest to 26.80 +/- 6.56 after OCC and 32.80 +/- 8.26ml/100mltissue following ROCC in Type I diabetic subjects. There were no differences in forearm blood flow between groups for any of the conditions. These data indicate the degree of forearm blood flow is directly related to the intensity of the vasodilatory stimulus. However, our study did not reveal evidence of impaired vasodilatory capacity in Type I diabetic subjects compared to controls in the absence of other risk factors.

Adult↗
PubMed2001

Promoting breast and cervical cancer screening at the workplace: results from the Woman to Woman Study.

OBJECTIVES: This article reports findings from a peer-delivered intervention designed to increase use of breast and cervical cancer screening. METHODS: Twenty-six worksites were randomly assigned to the intervention or comparison group. The 16-month intervention consisted of group discussions, outreach, and educational campaigns. Data were collected from a random sample of women employees stratified by age (baseline n = 2943; final n = 2747). Cross-sectional analyses were conducted to evaluate the impact of the intervention on screening behaviors. RESULTS: Relative to comparison worksites, the intervention group experienced greater increases in the percentage of women who reported a recent mammogram (7.2% vs 5.6%), clinical breast examination (5.8% vs 2.1%), and Papanicolaou (Pap) test (4.7% vs 1.9%). After worksite cluster and age strata were controlled for, the observed increase in Pap tests was significantly greater in the intervention group (odds ratio [OR] = 1.28; 95% confidence interval [CI] = 1.01, 1.62); however, differences in mammography screening rates (OR = 1.14; 95% CI = 0.90, 1.44) and clinical breast examination (OR = 1.19; 95% CI = 0.96, 1.49) were not statistically significant. CONCLUSIONS: Intervention activities produced a modest increase in cervical cancer screening, but they did not accelerate breast cancer screening rates above the observed secular trend.

Adult↗
PubMed2000

Extensive contribution of the multidrug transporters P-glycoprotein and Mrp1 to basal drug resistance.

Despite accumulating evidence that multidrug resistance transporter proteins play a part in drug resistance in some clinical cancers, it remains unclear whether the relatively low levels of multidrug resistance transporter expression found in most untreated tumors could substantially affect their basal sensitivity to antineoplastic drugs. To shed light on this problem, the drug sensitivities of wild-type mouse cell lines were compared with those of lines in which the Mdr1a and Mdr1b genes encoding P-glycoprotein (P-gp) were inactivated and lines in which the Mrp1 gene was inactivated in addition to Mdr1a and Mdr1b. These models permit a clean dissection of the contribution of each transporter to drug resistance at expression levels similar to those in normal tissues and avoid complications that might arise from previous exposure of cell lines to drug selection. For substrate drugs, we found that these contributions can indeed be very substantial. Lines lacking functional P-gp were, on average, markedly more sensitive to paclitaxel (16-fold), anthracyclines (4-fold) and Vinca alkaloids (3-fold). Lines lacking both P-gp and Mrp1 were (compared with wild-type lines) hypersensitive to an even broader array of drugs, including epipodophyllotoxins (4-7-fold), anthracyclines (6-7-fold), camptothecins (3-fold), arsenite (4-fold) and Vinca alkaloids, especially vincristine (28-fold). Thus, even very low levels of P-gp and Mrp1 expression that may be difficult to detect in tumors could significantly affect their innate sensitivity to a wide range of clinically important substrate drugs. An implication is that the use of resistance reversal agents to sensitize drug-naive tumors may be appropriate in more cases than is presently appreciated.

3T3 Cells↗
PubMed2000

Influence of age and normal plasma fibrinogen levels on flow-mediated dilation in healthy adults.

This study examined the effects of age-associated increases in fibrinogen on brachial artery flow-mediated dilation in 30 healthy nonsmokers. The findings demonstrate an inverse association between normal plasma fibrinogen levels and vascular function (r = -0.56, p = 0.001), suggesting elevated plasma fibrinogen may decrease the artery's responsiveness to certain vasodilatory signals, such as shear stress.

Adult↗
PubMed2000

Breast cancer resistance protein is localized at the plasma membrane in mitoxantrone- and topotecan-resistant cell lines.

Tumor cells may display a multidrug resistant phenotype by overexpression of ATP-binding cassette transporters such as multidrug resistance (MDRI) P-glycoprotein, multidrug resistance protein 1 (MRP1), and breast cancer resistance protein (BCRP). The presence of BCRP has thus far been reported solely using mRNA data. In this study, we describe a BCRP-specific monoclonal antibody, BXP-34, obtained from mice, immunized with mitoxantrone-resistant, BCRP mRNA-positive MCF-7 MR human breast cancer cells. BCRP was detected in BCRP-transfected cells and in several mitoxantrone- and topotecan-selected tumor cell sublines. Pronounced staining of the cell membranes showed that the transporter is mainly present at the plasma membrane. In a panel of human tumors, including primary tumors as well as drug-treated breast cancer and acute myeloid leukemia samples, BCRP was low or undetectable. Extended studies will be required to analyze the possible contribution of BCRP to clinical multidrug resistance.

ATP Binding Cassette Transporter, Subfamily G, Mem↗
PubMed2000

Deletion of cysteine 369 in lysyl hydroxylase 1 eliminates enzyme activity and causes Ehlers-Danlos syndrome type VI.

This study describes the relative contribution of the 10 cysteine residues in lysyl hydroxylase 1 (LH1) to enzyme activity. We have identified a novel mutation of a 15-bp deletion in exon 11 in one LH1 allele, that codes for amino acids 367-371 (DLCRQ), in two unrelated compound heterozygous patients with Ehlers-Danlos type VI. The mutations in their other alleles were a C1119T change (exon 10) and a predicted Q49X (exon 2). We confirmed that the loss of cysteine 369 in the deleted sequence contributed to the diminished enzyme activity by structure/function analysis of mutant LH1 constructs, in which C369 and the nine other cysteines were individually mutated to serine by site-directed mutagenesis of a normal pAcGP67/LH1cDNA construct. Following their expression in an Sf9 insect cell/baculovirus system, SDS-PAGE and Western analysis showed that equivalent levels of correctly-sized (85-kDa) products were secreted. The mutation of residues C369 and also C375, C552 and C687 virtually eliminated LH activity, whereas mutations of C267, C270, and C680 had an intermediate effect. In contrast, the C204S, C484S and C566S constructs had normal activity. Although disulfide bond formation may affect the relative contribution of each cysteine to LH activity, catalytic activity does not appear to be directly related to dimerization of the enzyme.

Alleles↗
PubMed2000

Inotropic response to endothelin-1, isoprenaline and calcium in cardiomyocytes isolated from endotoxin treated rats: effects of ethyl-isothiourea and dexamethasone.

1. The contractile effects of endothelin-1, isoprenaline and extracellular calcium were assessed on ventricular cardiomyocytes isolated from lipopolysaccharide-treated rats. The involvement of nitric oxide was investigated using dexamethasone (in vivo) and ethyl isothiourea (in vitro). 2. Male Wistar rats (n=70) were injected with either saline (1 ml kg(-1)) or lipopolysaccharide (LPS; 5 mg kg(-1)) alone, or following pre-treatment with dexamethasone (DEX+LPS; 5 mg kg(-1)). Ventricular cell shortening was recorded using a video edge detection system, and concentration-response relationships were established for endothelin-1, isoprenaline and calcium, in the absence or presence of ethyl isothiourea (ETU; 10 microM). iNOS expression was assessed using reverse transcription-polymerase chain reaction. 3. iNOS mRNA expression was greater (P<0.001) in the LPS (iNOS/GAPDH ratio: 0.90+/-0.09) treated group compared to saline (iNOS/GAPDH ratio: 0.36+/-0.02). Baseline contractile amplitude was reduced (P<0.05) in the LPS (7.3+/-0.2 microm) and DEX+LPS groups (6.7+/-0.3 microm) compared to saline (8. 0+/-0.2 microm). 4. The concentration-dependent contractile response to endothelin-1 was attenuated (P<0.05) in the LPS group compared to saline (maximum change: 0.45+/-0.2 vs 1.8+/-0.2 microm). Neither ETU nor dexamethasone improved contractile function in the LPS-treated animals. 5. The concentration-dependent increase in the contractile response to isoprenaline was attenuated in the LPS-treated group compared to saline (P<0.05; maximum change: 1.7+/-0.4 vs 3.1+/-0.4 microm). This effect was reversed by ETU (maximum change: 3.7+/-0.6 microm). Pre-treatment with dexamethasone prevented a significant fall in contraction amplitude (maximum change: 2.4+/-0.4 microm). 6. The contractile response to calcium was reduced (P<0.05) in the LPS group compared to saline (maximum change: 8.7+/-0.6 vs 10.7+/-0.8 microm). Neither ETU nor dexamethasone restored contractile function in the LPS-treated group. 7. In conclusion, a nitric oxide-mediated inhibitory pathway is not responsible for the diminished contractile response to either endothelin-1 or extracellular calcium, but contributes to the hyporesponsiveness to isoprenaline in lipopolysaccharide treated rats.

Animals↗
PubMed1999

Hybrid (BDBB) interferon-alpha: preformulation studies.

A number of techniques, including RP-HPLC, HP-SEC and SDS-PAGE have been used in the delineation of degradative mechanisms of recombinant hybrid (BDBB) interferon-alpha (IFN-alpha) in the solution phase. Different degradation profiles are found according to medium pH. At pH 4.0 the major routes of degradation are via chemical transformation of the monomeric protein to a species which retains antiviral activity, and by self-proteolytic hydrolysis. At pH 7.6, methionine-oxidation is the major chemical degradative process. Protein aggregation is also a significant route of degradation at the higher pH. The results have assisted in a targeted preformulation screen of potentially stabilising excipients and possible parenteral solution dosage forms have been identified. Preliminary 'real-time' storage data confirm excellent chemical and physical stability of IFN-alpha in vehicles formulated at pH 7.6 or, especially, pH 4.0 under the proposed shelf conditions.

Chemistry, Pharmaceutical↗
PubMed1999

The mouse Bcrp1/Mxr/Abcp gene: amplification and overexpression in cell lines selected for resistance to topotecan, mitoxantrone, or doxorubicin.

Mouse fibroblast cell lines lacking functional Mdr1a, Mdr1b, and Mrp1 genes were selected for resistance to topotecan, mitoxantrone, or doxorubicin. Each of the resulting drug-resistant lines showed marked gene amplification of Bcrp1, the mouse homologue of the human ATP-binding cassette transporter gene BCRP/MXR/ABCP, and greatly elevated expression of Bcrp1 mRNA. All three of the resistant cell lines were highly cross-resistant to topotecan and mitoxantrone and, to a variable extent, doxorubicin. All showed greatly reduced cellular accumulation and greatly increased efflux of mitoxantrone that was dependent on cellular ATP and efficiently reversed by the compound GF120918. The mouse Bcrp1 cDNA encodes a 657-amino-acid protein with 81% identity (86% similarity) to the human breast cancer resistance protein (BCRP) and a virtually superimposable hydrophobicity profile. Our data argue strongly that mouse Bcrp1 is functionally comparable with human BCRP, conferring multidrug resistance to topotecan, mitoxantrone, doxorubicin, and related compounds. Mouse models and cell lines should, therefore, be highly informative in understanding the clinical, pharmacological, and physiological roles of BCRP.

3T3 Cells↗
PubMed1999

Beliefs about Papanicolaou smears and compliance with Papanicolaou smear follow-up in adolescents.

OBJECTIVE: To explore qualitatively adolescent girls' understanding of Papanicolaou smears and barriers to compliance with Papanicolaou smear follow-up appointments. DESIGN: Qualitative analysis, using 3 focus groups and 15 in-depth, semistructured individual interviews. SETTING: Adolescent Clinic and Young Parents' Program at Children's Hospital, Boston, Mass. MAIN OUTCOME MEASURES: Beliefs and attitudes about Papanicolaou smears and barriers to compliance with Papanicolaou smear follow-up. RESULTS: The mean (+/- SD) age of the 15 interview participants was 18.7 (+/- 1.9) years. Knowledge about Papanicolaou smears and pelvic examinations was poor. Most participants believed that their peers receive Papanicolaou smear screening and perceived teenagers to be susceptible to cervical cancer. Perceived benefits to getting Papanicolaou smears were prevention and early detection or diagnosis, and reported barriers included pain or discomfort, embarrassment, fear of finding a problem, fear of the unknown, denial, poor communication or rapport with the provider, not wanting to look for trouble, lack of knowledge, and peers' advice. Participant-generated strategies for how providers could overcome barriers to Papanicolaou smear screening included education and the development of trusting, consistent relationships with providers. Participant-generated strategies for how providers could enhance appointment-keeping among adolescents included telephone and written reminders. CONCLUSIONS: These data support a behavioral theory-based model of adolescent compliance with Papanicolaou smear follow-up, which may help to develop strategies to enhance compliance with Papanicolaou smear follow-up appointments. These strategies include providing in-depth education about Papanicolaou smears, addressing barriers to Papanicolaou smear follow-up, focusing on appropriate provider behaviors, and instituting an appointment reminder system.

Adolescent↗
PubMed1999

The relationship between social network characteristics and breast cancer screening practices among employed women.

This study examined the relationship between social network characteristics and breast cancer screening practices among employed women. We hypothesized that larger social networks, higher levels of support from networks, and stronger social influences to undergo screening would be positively associated with regular utilization of mammograms and clinical breast examinations. Data were collected from women aged 52 and over who were employed in 27 worksites (N = 1,045). Social network characteristics, breast cancer screening practices, and sociodemographic factors were assessed in a self-administered survey. Bivariate analyses revealed that social influences were significantly associated with regular screening; social support was only marginally associated with regular screening; and social network size was not at all associated. In multivariate analyses, only the perception that screening is normative among one's peers was predictive of regular screening. Provider recommendation was the single most potent predictor of regular screening. These findings provide support for the importance of social norms in motivating women to adhere to screening guidelines. In addition, they underscore the potent impact of provider recommendations on women's screening practices.

Aged↗
PubMed1998

Topographic differences in CNV amplitude reflect different preparatory processes.

Topographic differences in Contingent Negative Variation (CNV) were recorded while people were preparing for cognitive versus motor tasks in an S1-S2 paradigm. CNV had a frontal distribution when people prepared to encode words into long-term memory, whereas CNV was more centrally distributed when the tasks were predominantly motoric. These topographic differences appeared to be related to the type of task rather than the amount of information extracted from the S2, because a direct manipulation of the level of S2 processing had little effect on CNV amplitude. The topographic differences in CNV suggest that preparation for motor activity is a different psychological process from preparation for stimulus processing and that these two processes are subserved by different neural structures. This experiment also demonstrated that a recognition memory paradigm can be useful in the investigation of the psychological correlates of CNV.

Adolescent↗