Protoporphyrin biosynthesis in Melanoma B16 cells stimulated by 5-aminolevulinic acid and chemical inducers: Characterization of photodynamic inactivation

The stimulation of protoporphyrin (PP) biosynthesis in BI6 melanoma cells in order to facilitate photodynamic cell killing was studied. Biosynthesis and accumulation of PP in the melanoma cells was increased from 8 to 15 pmol/mg protein by the use of dimethyl-sulfoxide (DMSO), a differentiation-inducer. Treatment of the cells with the porphyrogenic agent alh/l-isopropyl-acetamide (AIA) stimulated an additional PP increase. The most remarkable enhancement of intracellular PP was achieved by the supplementation of 5-aminolevulinic acid (5-ALA) to the growth medium following the addition of DMSO and AIA during the induction phase. The intracellular concentration of PP exceeded 21 950 pmol/mg protein following combined stimulation by DMSO/AIA and 5-ALA. The porphyrins produced in the incubated cells, in serum-depleted medium, consisted of 95% PP; 88% of it was recovered from the cells and only 7% was excreted into the medium. Photosensitization of the B16 melanoma cells containing high PP concentrations was effective even at low light doses. Potassium (K) efflux was the first measurable sign of cell damage determined by X-ray microanalysis (XRMA) following fast liquid-nitrogen fixation. During a I min interval, 70% of cellular K was lost. After 5 min illumination, complete cell destruction was detected by scanning electron microscopy (SEM) and XRMA. The photodam-aged cells showed influx of Na, CI and Ca ions accompanying the immediate K losses. Ultrastructural cell damage was manifested by disintegration of the outer membrane. Total cell death of B16 melanoma cells was achieved by chemical induction of endogenous PP and photosensitization.     

The Allergy Pricker

Skin prick test has advantages over other diagnostic tests in allergy, and attempts to increase the reproducibility are warranted. A standardised disposable precision needle with a needle point of 1.0 mm has recently become available. Based on 960 tests with histamine and grass pollen ( Phleum pratense ) the reproducibility of the skin prick weal area was calculated. Using histamine 10 mg/ml and grass pollen (1000 and 5000 PNU) A significantly lower coeffcient of variation was found compared with a stadard blood lancet with a point of 4 mm. The mean weal reaction is reduced to about 80% of the size obtained with the blood lancet. Testing with the precision needle resulted in a significantly reduced incidence of bleedings. The precision needle simplifies the skin test and does not require as much skill as other needles, and is recommended for both routine tests and the biological standardization of allergen extracts.     

Fistulizing reoperations in open-angle glaucoma

The authors analyze the results of repeated fistulization surgery (traditional trabeculectomy and that combined with resection of the episclera, and valvular trabeculectomy) in 40 patients (41 eyes) with primary open-angle glaucoma. A follow-up of 6 mos to 4 yrs has shown normalization of the ophthalmic tone in 90.2% of cases, in 65.8% of these this was achieved without drugs. Vision acuity remained unchanged or improved in 58.5% of cases, visual field was preserved or improved in 80.5% of patients. These results have lead the authors to a conclusion that a stable hypotensive effect can be achieved if repeated fistulization is performed outside the site of the primary intervention. If repeated trabeculectomy has to be performed at the site of the primary intervention, it is recommended to be combined with resection of the episclera, this improving the reliability of the hypotensive effect of surgery.     

Representativity of a postal public health questionnaire survey in Sweden, with special reference to ethnic differences in participation

AIM: Non-participation in health surveys is a common phenomenon. When differences between participants and non-participants are considerable, the external validity of the sample survey may decrease and false conclusions might be drawn about the health status of the population. For this reason, the authors aimed to investigate the representativity of a postal questionnaire survey performed in the county of Scania, Sweden, in 1999-2000. The survey, which was based on an 18- to 80-year-old population sample, had a 58% response rate (n = 13 604). METHODS: For some variables, the information obtained using the questionnaire was compared with information obtained from a population register that covers all the population in the county (for the 18- to 80-year-old group, n = 850 476). The population register includes, among other data, information on age, gender, educational level, country of birth, and healthcare expenditure. RESULTS: Men, individuals with a low level of education, and immigrants were under-represented in the survey. However, except for immigrants, the under-representation was not large. Among immigrants, particularly those born in former Yugoslavia, the Arabic-speaking countries, and Poland were very significantly under-represented in the study. By contrast, immigrants born in other Nordic countries had responded to almost the same extent as respondents born in Sweden. The survey sample had about the same healthcare utilization costs as did the general population. CONCLUSIONS: In summary, the "Health Survey for Scania, 2000" seems largely representative of the total Scanian population. A major concern, however, is the under-representation of the immigrant population.     

Salivary cortisol and self-reported stress among persons with environmental annoyance

OBJECTIVES: Increased vulnerability to stress has been suggested as a possible mechanism behind medically unexplained conditions such as sensitivity to electricity and common smells. This study examined whether subjective environmental annoyance among the general population is associated with increased physiological reactivity or subjective stress scores. METHODS: Four groups were studied (N=141): an electrically annoyed (N=17), a smell-annoyed (N=29), and a generally annoyed group (N=39) and a reference group matched for age, gender, and socioeconomic status (N=56). Over 5 days, the participants collected saliva for cortisol determination at awakening, 30 minutes after awakening, 8 hours after awakening, and at 9 o'clock in the evening. On the evening preceding the fifth day, the participants ingested a 0.5-mg dexamethasone tablet so that possible differential suppression of the hypothalamic-pituitary-adrenal (HPA) axis could be assessed. Each day, the participants also rated their subjective stress and health complaints. RESULTS: No significant differences were found between the groups regarding cortisol secretion over 5 days. The dexamethasone suppression test showed inhibited cortisol secretion in all four groups. No associations were found between the cortisol concentrations and the self-reported stress scores or subjective health complaints. CONCLUSIONS: Although the environmentally annoyed groups showed no signs of increased HPA-axis activation, being annoyed by both electrical devices and smells seems to be related to increased psychological activation in terms of self-reported stress. Because the participants were otherwise healthy and recruited from the general population, the results imply that subtle psychological stress processes may be important in the early development of environmental annoyance.     

Neuroblastoma cell death in response to docosahexaenoic acid: sensitization to chemotherapy and arsenic-induced oxidative stress

Docosahexaenoic acid (DHA) is an omega-3 polyunsaturated fatty acid vital for the developing nervous system and significantly decreased in neuroblastoma cells compared to nontransformed nervous tissue. We investigated whether supplementation of DHA affects the susceptibility of neuroblastoma cells to oxidative stress generated endogenously and in response to cytotoxic therapy. DHA, but not the monounsaturated oleic acid (OA), induced dose- and time-dependent neuroblastoma cell death. DHA supplementation was associated with depolarization of the mitochondrial membrane potential, production of reactive oxygen species (ROS) and accumulation of DNA in sub-G1 phase of the cell cycle. The antioxidant, vitamin E, inhibited mitochondrial depolarization and subsequent cell death induced by DHA, whereas, the mitochondrial pore inhibitor, cyclosporin A, partly inhibited DHA-induced neuroblastoma cell death. Depletion of glutathione by L-buthionine-sulfoximine significantly enhanced the cytotoxic effects of DHA. Nontransformed fibroblasts were not substantially affected by DHA. DHA, but not OA, significantly enhanced the cytotoxicity of cisplatin, doxorubicin and irinotecan both in chemosensitive and in multidrug-resistant neuroblastoma cells. DHA potently sensitized neuroblastoma cells to a clinically relevant concentration (1 microM) of arsenic trioxide (As2O3) and enhanced the effect of the nonsteroidal antiinflammatory drug (NSAID), diclofenac. These findings provide experimental evidence that the omega-3 fatty acid, DHA, is cytotoxic to drug-resistant neuroblastoma. The potent action of DHA with arsenic trioxide, NSAID and chemotherapeutic agents suggests clinical testing of this therapeutic concept in children with neuroblastoma.