Biomarkers for exposure to ambient air pollution--comparison of carcinogen-DNA adduct levels with other exposure markers and markers for oxidative stress

Human exposure to genotoxic compounds present in ambient air has been studied using selected biomarkers in nonsmoking Danish bus drivers and postal workers. A large interindividual variation in biomarker levels was observed. Significantly higher levels of bulky carcinogen-DNA adducts (75.42 adducts/10(8) nucleotides) and of 2-amino-apidic semialdehyde (AAS) in plasma proteins (56.7 pmol/mg protein) were observed in bus drivers working in the central part of Copenhagen, Denmark. In contrast, significantly higher levels of AAS in hemoglobin (55.8 pmol/mg protein), malondialdehyde in plasma (0. 96 nmol/ml plasma), and polycyclic aromatic hydrocarbon (PAH)-albumin adduct (3.38 fmol/ microg albumin) were observed in the suburban group. The biomarker levels in postal workers were similar to the levels in suburban bus drivers. In the combined group of bus drivers and postal workers, negative correlations were observed between bulky carcinogen-DNA adduct and PAH-albumin levels (p = 0.005), and between DNA adduct and [gamma]-glutamyl semialdehyde (GGS) in hemoglobin (p = 0.11). Highly significant correlations were found between PAH-albumin adducts and AAS in plasma (p = 0.001) and GGS in hemoglobin (p = 0.001). Significant correlations were also observed between urinary 8-oxo-7, 8-dihydro-2'-deoxyguanosine and AAS in plasma (p = 0.001) and PAH-albumin adducts (p = 0.002). The influence of the glutatione S-transferase (GST) M1 deletion on the correlation between the biomarkers was studied in the combined group. A significant negative correlation was only observed between bulky carcinogen-DNA adducts and PAH-albumin adducts (p = 0.02) and between DNA adduct and urinary mutagenic activity (p = 0.02) in the GSTM1 null group, but not in the workers who were homozygotes or heterozygotes for GSTM1. Our results indicate that some of the selected biomarkers can be used to distinguish between high and low exposure to environmental genotoxins.     

Comparison of two 3-day Helicobacter pylori eradication regimens with a standard 1-week regimen

BACKGROUND: The duration of Helicobacter pylori eradication regimens has decreased to 1 week with cure rates of over 90%. This can be attributed to the use of triple drug regimens including potent inhibitors of gastric acid secretion and clarithromycin. There is no theoretical reason why shorter regimens should not be possible. AIM: To compare two 3-day, low-dose, twice daily regimens with 1 week of omeprazole 20 mg b.d., clarithromycin 250 mg b.d., and metronidazole 400 mg b.d. (OCM) METHODS: Outpatients referred for gastroscopy were screened by biopsy urease test. H. pylori-positive patients were randomized to receive either lansoprazole 30 mg b.d., tri-potassium dicitrato bismuthate one tablet b.d., clarithromycin 250 mg b.d., and amoxycillin 1 g b.d. for 3 days (LTdbCA), or ranitidine bismuth citrate 400 mg b.d., clarithromycin 250 mg b.d. and amoxycillin 1 g b.d. for 3 days (RbcCA) or omeprazole 20 mg b.d., clarithromycin 250 mg b.d. and metronidazole 400 mg b.d. for 1 week (OCM). They were not pre-treated with a gastric acid inhibitor. After 8 weeks, H. pylori status was assessed by 13C urea breath test. RESULTS: 974 out of 1114 patients referred for gastroscopy were screened by biopsy urease test. 140 patients were not screened either because they were anticoagulated or for technical reasons. 334 patients were H. pylori-positive: 154 were excluded mostly because of allergy to penicillin and personal reasons but 180 were randomized to treatment All regimens were well tolerated. For LTdbCA (n=60), RbcCA (n=59), and OCM (n=61) the H. pylori cure rates (95% CI) were 23% (12-34), 14% (5-23) and 87% (79-95), respectively, using intention-to-treat analysis and 25% (14-36), 15% (6-24) and 88% (80-96), respectively, if analysed per protocol. OCM was significantly superior to LTdbCA and RbcCA (P < 0.001) but there was no significant difference between regimens LTdbCA and RbcCA. CONCLUSIONS: OCM is an extremely effective H. pylori eradication regimen. The 3-day regimens tested both have poor cure rates. Pre-treatment with a proton pump inhibitor, higher doses or more frequent dosing may be necessary to increase the cure rate of short duration regimens. However, this could make them less acceptable than the H. pylori eradication regimens currently available.     

Combination of methotrexate and sulphasalazine in patients with rheumatoid arthritis: pharmacokinetic analysis and relationship to clinical response

1. The influence of sulphasalazine (SASP) on the pharmacokinetics of low dose methotrexate (MTX) and the relation between pharmacokinetic variables and clinical response was studied in 15 patients with active rheumatoid arthritis despite >6 months of SASP treatment.
2. SASP was stopped for 2 weeks. Thereafter a single oral dose of 7.5 mg MTX was administered after a standard breakfast. Blood was sampled initially every 30 min, thereafter hourly during 8 h. Urine was sampled every hour. Then 2000 mg SASP daily + 7.5 mg MTX weekly was given. After 4 weeks the same procedure was repeated supplemented with concomitant administration of 1000 mg SASP. Clinical measurements included Ritchie articular index, number of swollen joints, ESR and the disease activity score. Pharmacokinetic analysis was performed using a two- compartment model with first order absorption and lag time. Results are given as mean (s.d.). Paired t-test or signed rank test were applied in the statistical analysis.
3. Pharmacokinetics of MTX without vs with SASP, means±s.d. were as follows: AUC: 673±179 vs 628±210 (95% confidence interval [CI] of the difference was −71 to 159) ng ml⁻¹ h, MRT: 5.2±1.3 vs 5.2±1.1 (95% CI −0.4 to 0.4) h, t½,z: 4.3±1.1 vs 4.2±1.1 (95% CI −0.3 to 0.5) h, V /F: 59.3 ±29.3 vs 65.5±25.3 (95% -23.8 to 11.4) l, CL/F: 12.3±5.0 vs 13.5±4.8 (95% CI −4.5 to 2.3) l h⁻¹. CL_R/F: 6.2±1.3 vs 6.3±2.1 (95% CI −1.3 to 1.1) l h⁻¹. All P values were ≥0.3.
4. A weak correlation existed between the change of ESR and the MRT, the t½,z and the V /F (Spearman correlation coefficients of 0.43, 0.50 and 0.50 respectively, 0.05<P<0.1).
5. There is no significant influence of chronic SASP administration on the pharmacokinetics of MTX or vice versa. Of the clinical variables, only the ESR correlated consistently with some pharmacokinetic variables of MTX.

     

The mechanism of directional coronary atherectomy

An attempt was made to assess the mechanism of directional coronaryatherectomy using different methods of analysis. Quantitativecoronary angiography was used as the gold standard to assessthe immediate results of atherectomy, and a comparative quantitativeanalysis of atherectomy and balloon angioplasty was made. Todetermine whether the post-atherectomy cross-sectional areais close to a circle, we compared the area measurements obtainedby edge detection with those obtained by videodensitometry.Finally, the extent of a ‘Dotter’ effect was establishedby quantitative angiography following crossing the stenosiswith the atherectomy device. For the purpose of this study,the results of the first 113 successful atherectomy procedureswere reviewed. In matched lesions, directional atherectomy induceda greater increase in minimal luminal diameter than balloonangioplasty (1.6 mm vs 0.8 mm; P < 0.0001 However, this luminalimprovement is due to a substantial ‘Dotter’ effectinduced by the bulky atherectomy device. Following atherectomy,only a slight difference in cross-sectional area measurementsbetween edge detection and videodensitometry (mean difference:0.28 mm² was found. Histologic examination of an atherectomizedcoronary artery showed a near-circular post atherectomy areageometry. In conclusion, directional atherectomy is a very effectivedevice with a substantially better initial result than balloonangioplasty. However, insertion of this bulky device itselfcauses an important ‘Dotter’effect.     

CALCITONIN GENE-RELATED PEPTIDE II, SUBSTANCE P AND VASOACTIVE INTESTINAL PEPTIDE IN PLASMA AND SYNOVIAL FLUID FROM PATIENTS WITH INFLAMMATORY JOINT DISEASE

Immunoreactive plasma and synovial fluid concentrations of calcitoningene-related peptide II (CGRP II), substance P and vasoactiveintestinal peptide (VIP) were measured in patients with osteoarthritis,gout and rheumatoid arthritis. Significantly higher levels ofCGRP II and substance P and VIP-like immunoreactivity levelsin synovial fluid were found in gout as well as CGRP II, substanceP and VIP-like immunoreactivities in rheumatoid arthritis whencompared to those in osteoarthritis. Plasma CGRP II, substanceP and VIP-like immunoreactivity levels showed no significantdifferences among patients in the three different groups ofarthritis. Our results suggest that these neuropeptides releasedfrom peripheral nerve endings into the synovial cavity probablyplay a pathogenic role in human joint inflammation. KEY WORDS: Rheumatoid arthritis, Gout, Calcitonin gene-related peptide II, Substance P, Vasoactive intestinal peptide, Synovial fluid     

VOLTAGE FIELDS SURROUNDING NEEDLES USED IN REGIONAL ANAESTHESIA

Using a bench model, we have studied the voltage fields surroundingboth insulated and uninsulated needles used in regional anaesthesia.The findings were compared with earlier computer predictionswhich suggested that the fields would be markedly differentfor the two types of needle. The results confirm that the fieldsdiffer markedly and suggest that the use of insulated needlesmay not necessarily improve the accuracy of nerve location andthat uninsulated needles may be more appropriate. ^*Present addresses: Department of Anaesthetics, Hull Royal Infirmary,Hull, Humberside. Droitwich Knee Foundation, Saga House, SansomePlace, Worcester Present addresses: Droitwich Knee Foundation, Saga House, SansomePlace, Worcester.     

FUNCTIONAL ROLE OF CARDIAC PRESYNAPTIC α2-ADRENORECEPTORS IN THE BRADYCARDIA OF α-ADRENORECEPTOR AGONISTS IN PENTOBARBITONE- AND URETHANE-ANAESTHETIZED NORMOTENSIVE RATS

• 1 The relative potencies of 6 α-adrenoreceptor agonists, with differing physicochemical properties, at cardiac presynaptic α-adrenoreceptors were determined by measuring their ability to reduce the tachycardia produced by stimulation of the cardiac sympathetic nerves in pithed rats. The compounds studied were clonidine, B-HT 933 (azepexole), oxymetazoline, St 91, naphazoline and DPI.
• 2 The bradycardia produced by the same compounds in bilaterally vagotomized, urethane- or pento-barbitone-anaesthetized normotensive rats were also compared. The relative order of presynaptic potency appeared similar to that observed for the bradycardic activity of the compounds in urethane- and pento-barbitone-anaesthetized rats. In pentobarbitone-anaesthetized rats all compounds evoked a maximal effect of 18–20% reduction in heart rate. In urethane-anaesthetized rats, however, a difference was observed between clonidine and azepexole on the one hand and oxymetazoline, St 91, naphazoline and DPI on the other hand. The former induced a 20–22% reduction in cardiac frequency, whereas the latter diminished heart rate by 10–16% only. In vagotomized, bilaterally adrenalectomized, urethane-anaesthetized rats, clonidine, St 91, naphazoline and azepexole evoked a 25% reduction in heart rate, whereas a 20% reduction was observed for DPI and oxymetazoline.
• 3 A radio-enzymatic determination of plasma catecholamines demonstrated that under urethane-anaesthesia plasma adrenaline concentrations were significantly elevated over the values observed in pentobarbitone-anaesthetized rats. This rise in plasma adrenaline was related to the amount of urethane used. In urethane-anaesthetized, bilaterally adrenalectomized rats, plasma adrenaline was not significantly elevated.
• 4 These findings demonstrate the involvement of cardiac presynaptic α₂-adrenoreceptors in the acute bradycardia, evoked by the α-adrenoreceptor agonist upon intravenous application to pentobarbitone-anaesthetized, normotensive rats. In urethane-anaesthetized rats, however, the functional role of the cardiac presynaptic α₂-adrenoreceptors may be obscured as a result of the high plasma adrenaline levels observed in these animals.

     

IMPAIRMENT BY NIFEDIPINE OF VASOPRESSOR RESPONSES TO STIMULATION OF POSTSYNAPTIC α2-ADRENORECEPTORS IN GANGLION-BLOCKED RABBITS. FURTHER EVIDENCE FOR THE SELECTIVE INHIBITION OF POSTSYNAPTIC α2-ADRENORECEPTOR-INDUCED PRESSOR RESPONSES BY CALCIUM ANTAGONISTS

• 1 After ganglionic blockade and bilateral vagotomy, vasopressor responses induced by activation of postsynaptic α₁- and α₂-adrenoreceptors were elicited in the intact circulatory system of rabbits.
• 2 The hypertensive effects of the selective stimulating agents methoxamine (α₁-agonist) and B-HT 920 (α₂-agonist) were effectively antagonized by the adrenoreceptor antagonists prazosin and yohimbine, respectively. These findings confirm the existence of two types of postsynaptic α-adrenoreceptors (α₁-and α₂-type) in vascular smooth muscle of rabbits.
• 3 The calcium antagonistic drug nifedipine did not affect the maximal increase in diastolic pressure brought about by methoxamine, whereas it strongly inhibited the hypertensive effects of B-HT 920.
• 4 It is concluded that this confirmation of the selective inhibition of postjunctional α₂-adrenoreceptor-mediated vasopressor responses by a calcium antagonistic drug, such as nifedipine, indicates that this activity constitutes a general phenomenon. This finding supports the hypothesis that an influx of extracellular calcium is necessary for the vasoconstriction mediated by postsynaptic α₂-adrenoreceptors.

     

Neoadjuvant cisplatin plus ifosfamide in patients with stage IIB cervical cancer: a single center phase II study

The objective of this study was to evaluate cisplatin plus ifosfamide as neoadjuvant chemotherapy with regard to toxicity and clinical response in patients with stage IIB cervical cancer. Sixty-eight patients with previously untreated stage IIB cervical cancer were given two cycles of chemotherapy: cisplatin 20 mg m⁻² on Days 1–5, infused over 1 h; ifosfamide 1.2 g m⁻² on Days 1–5 infused over 30 min. Mesna 120 mg m⁻² was administered as a bolus 15 min before ifosfamide, and a continuous infusion, delivering Mesna 1.2 g m⁻², was given subsequently over the next 16 hours. The treatment cycle was repeated on day 21. Responders were then randomized to surgery or radiation therapy. All 68 patients were evaluable for toxicity. Toxicity was found to be acceptable. One patient died at home one month after completion of the second treatment cycle. There was one grade 4 thrombocytopenia. Grade 3 toxicities included anemia in four patients, leucopenia and nausea and vomiting in one patient each. Sixty-two patients were evaluable for response. A clinical response was documented in 44 of the 55 evaluable patients (80%), with 17 complete responses (31%) and 27 partial responses (49%) (95% confidence limits 69%–91%, 19%–43%, and 36%–62% respectively). The intent-to-treat response rate was 64.7%. Twenty-one patients were randomized to surgery and 23 patients to radiation therapy. Amongst the eight patients with a complete clinical response, one patient had a complete pathological response and one patient had residual intra-epithelial neoplasia. The drug combination of cisplatin plus ifosfamide had acceptable toxicity and gave a clinical response rate of 80% in previously untreated patients with stage IIB cervical cancer.     

Effect of the viable-yellow (A(vy)) agouti allele on skin tumorigenesis and humoral hypercalcemia in v-Ha-ras transgenic TGxAC mice

We previously reported that papillomas can arise from the follicular epithelium of v-Ha-ras transgenic TGxAC mice. Since the viable-yellow mutation (A(vy)) of the mouse agouti gene which regulates coat color pigmentation by acting within the micro-environment of the hair follicle has been shown to function as a tumor promoter in the liver, we hypothesized that it may also play a role in TGxAC skin tumorigenesis. Endogenous agouti protein product was detected in the outer root sheath of anagen hair follicles following plucking of the hair shaft, but not in the interfollicular epithelium, in TGxAC mice on an FVB/N genetic background. It was also detected in papillomas from these mice produced by 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment or plucking. Expression of the A(vy) allele in the v-Ha-ras transgenic TGxAC mouse line results in an approximately 2-fold increase in papilloma development compared with controls which did not carry the A(vy) allele following twice-weekly treatment with 1.25, 2.5 or 5.0 microg TPA. In addition, TPA-treated, papilloma-bearing F1 mice which carried the A(vy) allele, but not F1 mice which did not carry the A(vy) allele, exhibited a syndrome of humoral hypercalcemia mediated by parathyroid hormone-related protein (PTHrP) that led to weight loss, hypercalcemia and hypophosphatemia. Thus, we conclude that the A(vy) allele can influence the development of skin tumors and PTHrP-mediated humoral hypercalcemia in v-Ha-ras transgenic TGxAC mice.