Grafting assay distinguishes promotion sensitive from promotion resistant JB6 cells

The JB6 mouse epidermal cell system has been used extensively as an in vitro transformation model for the study of tumor promotion. The standard JB6 cell assay for promotion of transformation is carried out in soft agar or other anchorage independent conditions. The present study was directed to the development of an in vivo model to distinguish the promotion resistant (P-) and promotion sensitive (P+) progression phenotypes. Results indicate that the grafting assay distinguishes P- and P+ cells in vivo with P+ but not P- cells forming tumors within 7-9 weeks. Expression of dominant negative mutant jun TAM67 blocks both anchorage independent transformation response and graft bed tumor formation by P+ cells, suggesting that the requirement for AP-1 activation in transformation now applies in vivo. Expression of mutated p53 produced a gain of P+ phenotype in P- cells in vitro, but not in vivo. Histochemical and Northern blot analysis for expression of various keratinocyte markers revealed no evidence for expression, suggesting a loss of keratinocyte markers following establishment in culture. In summary, the skin-grafting assay described in this study appears to be a valid in vivo assay for distinguishing the preneoplastic progression phenotypes represented by JB6 P- and P+ cells.      

Glycopyrrolate treatment of drooling in an adult male patient with cerebral palsy

1. The aim of the study was to assess the effect of glycopyrrolate on drooling in an adult male patient with cerebral palsy. 2. After a thorough medical examination and consent in writing by the responsible guardian, a baseline sum score for frequency and seriousness of drooling was established over a 2 week period in addition to data on shifts of handkerchiefs, urination, defecation and observation of behaviour. Glycopyrrolate (1 mg) tablets were then administered, starting with one tablet daily the third week and increasing the daily dose by one tablet per week until a maximum of four tablets during week six and 4 days of week seven when the daily dose was reduced to two tablets for 3 days. For the four weeks 8-11 three tablets were given daily. In week 12 the dose was reduced to two tablets and for the weeks 13-15 no tablets of glycopyrrolate were given. 3. For as long as the patient received three to four tablets of glycopyrrolate daily, drooling was markedly reduced and handkerchiefs were not necessary on some days. After the tablets were withdrawn drooling increased to approximately the same level as it was before treatment. No adverse medical, psychological, or social effects were observed. 4. For shorter periods, glycopyrrolate can be given in controlled doses provided that an adequate medical assessment has been undertaken.     

Development of oxygen sensitivity in infants of smoking and non-smoking mothers

AIMS: To assess the effect of prenatal cigarette smoke exposure on the postnatal resetting of oxygen sensitivity in term infants. METHODS: 15 healthy term infants of smoking mothers (median 10 cigarettes/day) and 16 controls were studied during quiet sleep 1, 3, and 10 days and 10 weeks postnatally. Strain-gauge respiratory trace was continuously recorded. Repeated 15-s challenges with 100% O2 and 15% O2 were presented in randomised order through a face mask. A median of six hyperoxic and six hypoxic challenges per recording were obtained. Breath-by-breath ventilation in a time-window from 20 s before onset of stimulus to 60 s after was extracted. For each infant at each age, the normalised coherently averaged response to hyperoxia and hypoxia was calculated. Mean ventilation at end of the 15-s stimulus was analysed with ANOVA, as were parameters describing a function fitted to each averaged response. RESULTS: During air breathing, smoke-exposed infants had higher respiratory rates and lower tidal volumes than controls. Nicotine concentration in infant hair, measured by gas chromatography, was positively correlated with maternal level of smoking. A long-term development in oxygen sensitivity was demonstrated in both groups. However, neither the time-course nor the magnitude of O2 responses was affected by maternal smoking. Overall, hyperoxia reduced ventilation by 6.3% at day 1, 13.2% at day 3, 29.6% at day 10, and 40.0% at week 10. Transient hypoxia increased ventilation by 3.5%, 3.2%, 6.4%, and 8.8%, respectively, at the four ages studied.     

Importance of overall treatment time for the response to radiotherapy in patients with squamous cell carcinoma of the head and neck

In the nineties, several national protocols by the Danish Head and Neck Cancer Study group DAHANCA and other randomized trials, which have included several thousand patients, were performed on squamous cell carcinoma of the head and neck, now the most common malignant disease worldwide. It is a locoregional disease and distant metastases are rarely seen at diagnosis. Radiotherapy and surgery are thus the treatment of choice, with radiotherapy being the treatment modality if organ conservation is required. Since the late eighties there has been a strong focus on the importance of overall treatment time for the outcome of curative radiotherapy for these carcinomas. Based on the results of the Danish protocols it was concluded that the schedule of radiotherapy should be given with the shortest possible overall treatment time. In fact, as a consequence of the loco-regional control rate, the disease-specific and overall survival have shown a significant dependency on the overall treatment time: when this is short, the most beneficial results are achieved. Furthermore, treatment with 6 fractions per week is now the standard radiotherapy in Denmark in most head and neck carcinomas, associated with hypoxic modification using nimorazole. The response to accelerated fractionation is however heterogeneous and until proper predictive factors can be identified and further clarified which are the patients who truly benefit from accelerated fractionation, the radiotherapy schedules should secure a sufficient dose to all patients.     

Plasma osteopontin, hypoxia, and response to the hypoxia sensitiser nimorazole in radiotherapy of head and neck cancer: results from the DAHANCA 5 randomised double-blind placebo-controlled trial

BACKGROUND: The concentration of osteopontin (SPP1) in plasma is associated with tumour hypoxia. The DAHANCA 5 trial found that the hypoxia radiosensitiser nimorazole significantly improved the outcome of radiotherapy for patients with head and neck cancer compared with placebo. However, whether all patients benefit from such modification of hypoxia is unclear. We aimed to assess whether the concentration of plasma osteopontin could predict response to the hypoxia radiosensitiser. METHODS: Plasma concentrations of osteopontin were measured by use of ELISA from stored samples of 320 patients randomised in the DAHANCA 5 trial. Samples were grouped into tertiles according to high (167-1382 microg/L), intermediate (69-166 microg/L), or low (0-68 microg/L) concentrations of plasma osteopontin, and analysed for locoregional tumour control and disease-specific survival at 5 years. FINDINGS: Overall, locoregional tumour failure and disease-specific mortality were more frequent in patients assigned placebo than in those assigned nimorazole (relative risk [RR] 0.51 [95% CI 0.32-0.79] and 0.54 [0.35-0.85], respectively). Locoregional tumour failure was more frequent in patients with high concentrations of osteopontin assigned placebo than in those with high concentrations assigned nimorazole (0.19 [0.08-0.44]), as was disease-specific mortality (0.25 [0.11-0.59]). However, neither locoregional tumour failure nor disease-specific mortality differed between groups for patients with low concentrations of plasma osteopontin (0.79 [0.26-1.70]) and (0.69 [0.31-1.51]) or for those with intermediate concentrations (0.90 [0.41-1.98] and 0.89 [0.41-1.96], respectively). INTERPRETATION: High plasma concentrations of osteopontin are associated with a poor outlook after radiotherapy for patients with head and neck cancer, but can be improved by use of nimorazole. High concentrations of osteopontin can predict clinically relevant hypoxia, and might identify patients who will benefit from modification of hypoxia during radiotherapy.     

Reduced release of vasoconstrictors from the porcine heart after repeated periods of ischemia

OBJECTIVE: To examine if the decline in post-ischemic hyperemic flow after repeated brief periods of myocardial ischemia is accompanied by augmented cardiac release of the vasoconstrictors endothelin-1 (ET-1) and norepinephrine (NE). DESIGN: Mid-LAD (left anterior descending coronary artery) was occluded for 10 min with 30 min intervals a total of four times in six anesthetized pigs. Blood from the anterior interventricular coronary vein was drained through a shunt to the right atrium to facilitate blood sampling. Plasma concentrations of ET-1 and NE were repeatedly measured in arterial and coronary venous blood to estimate cardiac vasoconstrictor release. RESULTS: Plasma concentrations of ET-1 and NE remained unaltered, but cardiac release of both vasoconstrictors rose briefly during reperfusion due to the hyperemia. However, release declined progressively after repeated periods of ischemia and reperfusion and amounted to 53% (NE) and 17% (ET-1) of initial release after the fourth period of ischemia. CONCLUSION: The decline in post-ischemic hyperemia after repeated brief periods of myocardial ischemia is not accompanied by a progressive accentuation of cardiac ET-1 and NE release.     

Glycaemic control in type II diabetic tube-fed patients with a new enteral formula low in carbohydrates and high in monounsaturated fatty acids: a randomised controlled trial

OBJECTIVES: To investigate the effects of long-term treatment with a new enteral formula low in carbohydrates and high in monounsaturated fatty acids (MUFAs), in comparison with a standard formula, on glycaemic control in tube-fed type II diabetic patients. DESIGN: Randomised, double-blind, controlled, multi-centre trial.Setting:Early rehabilitation centres, primary care and nursing facilities. SUBJECTS:A total of 78 patients with insulin-treated type II diabetes with HbA(1C) > or =7.0% and/or fasting blood glucose >6.66 mmol/l, who required enteral tube feeding due to neurological dysphagia. INTERVENTIONS: Patients received 113 kJ (27 kcal)/kg of body weight of either test feed or an isoenergetic, isonitrogenous enteral formula (control) for 12 weeks. Glycaemic control (total daily insulin dosage (IU), fasting blood glucose, and HbA(1C)) and gastrointestinal tolerance were monitored daily. RESULTS: After 12 weeks, median values for changes from baseline were as follows (test group vs control group, 'data as available' analysis): total daily IUs -6.0 vs 0.0 (P=0.0024), fasting blood glucose (mmol/l) -1.59 vs -0.08 (P=0.0068); HbA(1C) (%) -0.8 vs 0.0 (P=0.0016). Both formulas were tolerated comparably. CONCLUSIONS:This study indicates that in tube-fed insulin-treated type II diabetic patients, the new low-carbohydrate, high MUFA formula results in a more effective glycaemic control than the standard diet, while being comparable in safety.     

The effect of the market opening on trends in smoking rates in Taiwan

OBJECTIVES: To examine smoking rates before and after the opening of the market to foreign imports. METHODS: Consumer surveys of Monopoly Bureau, National Health Interview Survey and official tobacco consumption, production and import data were used to calculate smoking rates, cigarette consumption, and the market share of imports. Age-adjusted smoking rates were used to compare trends before and after the market opening. RESULTS: The market share of the imports reached half in 2001 from 2% before opening. Compared to projected rates, smoking rates in 2001 for all and younger males, and all and younger females were increased by 12, 6, 202, and 249%, respectively, over and above the projected trends. The ratio of smoking rates between younger and older adults became larger, from 0.62 to 0.74 in males and from 0.32 to 0.98 in females. Per capita consumption increased after the market opening, particularly when large increases in smuggled cigarettes were considered. CONCLUSIONS: The finding that the market opening triggered an increase in smoking rates is contrary to the assertion by U.S. cigarette producers that importation would only make smokers switch brands. Younger adults and females were more affected, reflecting their higher sensitivity to the appeal from foreign cigarettes. Because of the marketing strategy of imports, more young people smoked, at an earlier age. Experience from Taiwan revealed that after the initial increase in smoking rates, the opening had galvanized the anti-smoking sentiments, legitimized and strengthened tobacco control policies and tempered the severity of the adverse impact.     

Successful nonsibling bone marrow transplantation in severe combined immunodeficiency.

Severe combined immunodeficiency (SCID) was diagnosed in a girl immediately after birth; her older brother had SCID and was successfully reconstituted by bone marrow transplantation from his uncle. She was isolated in a laminar air flow bench and decontaminated. The father differed by one HLA-A antigen but was HLA-Dw2 homozygous like the patient; his lymphocytes showed a slight response to the patient's cells in mixed lymphocyte culture (MLC). At the age of 2 1/2 months and again at 5 months, she was given a bone marrow transplant from the father. During the entire course the patient had no infections, and apart from a transient eosinophilia she had no signs of graft-versus-host reaction. Immunological reconstitution was nearly complete at 9 months of age, when she was recontaminated. One year later plasma immunoglobulin concentrations are in the low normal range (IgG and IgM) or decreased (IgA); tests of cell-mediated immunity are normal. Apart from slight upper respiratory infections, the patient has been healthy. Physical and psychological development have been normal.     

Extrinsic discoloration of teeth

The majority of tooth discolorations are extrinsic in nature and appear as brown integuments. Various clinical indices and photometric techniques have been used for the evaluation of extrinsic discolorations. Smoking, tea or coffee consumption and increasing age are promoting factors and such discolorations are frequently seen in connection with oral use of antibacterial plaque-inhibiting mouthrinses. Chemical alteration of the acquired pellicle appears to be the major reason for these brown integuments.