Use of cellular and cordless telephones and risk of testicular cancer.

A case-control study on testicular cancer included use of cellular and cordless telephones. The results were based on answers from 542 (92%) cases with seminoma, 346 (89%) with non-seminoma, and 870 (89%) controls. Regarding seminoma the use of analog cellular phones gave odds ratio (OR) = 1.2, 95% confidence interval (CI) = 0.9-1.6, digital phones OR = 1.3, CI = 0.9-1.8, and cordless phones OR = 1.1, CI = 0.8-1.5. The corresponding results for non-seminoma were OR = 0.7, CI = 0.5-1.1, OR = 0.9, CI = 0.6-1.4, and OR = 1.0, CI = 0.7-1.4, respectively. There was no dose-response effect and OR did not increase with latency time. No association was found with place of keeping the mobile phone during standby, such as trousers pocket. Cryptorchidism was associated both with seminoma (OR = 4.2, CI = 2.7-6.5) and non-seminoma (OR = 3.3, CI = 2.0-5.6), but no interaction was found with the use of cellular or cordless telephones.

Use of cellular telephones and brain tumour risk in urban and rural areas.

AIMS: To investigate the association between the use of cellular or cordless telephones and the risk for brain tumours in different geographical areas, urban and rural. METHODS: Patients aged 20-80 years, living in the middle part of Sweden, and diagnosed between 1 January 1997 and 30 June 2000 were included. One control matched for sex and age in five year age groups was selected for each case. Use of different phone types was assessed by a questionnaire. RESULTS: The number of participating cases was 1429; there were 1470 controls. An effect of rural living was most pronounced for digital cellular telephones. Living in rural areas yielded an odds ratio (OR) of 1.4 (95% CI 0.98 to 2.0), increasing to 3.2 (95% CI 1.2 to 8.4) with >5 year latency time for digital phones. The corresponding ORs for living in urban areas were 0.9 (95% CI 0.8 to 1.2) and 0.9 (95% CI 0.6 to 1.4), respectively. This effect was most obvious for malignant brain tumours. CONCLUSION: In future studies, place of residence should be considered in assessment of exposure to microwaves from cellular telephones, although the results in this study must be interpreted with caution due to low numbers in some of the calculations.

How to combine the use of different mobile and cordless telephones in epidemiological studies on brain tumours?

Mobile phone users in epidemiological studies have often used more than one phone model, and sometimes also more than one mobile phone system (analogue and digital systems). Until now, this has not been taken into account in epidemiological studies, mainly because we do not know the possible interaction mechanism(s) and, hence, how to integrate exposure from different phones into one dosimetric measure. In this paper we take a step towards starting a discussion about how to proceed with this important issue and the possible use of parameters such as weighting factors, measured specific absorption rate (SAR) values and integrated specific absorption values are discussed. As a base of this discussion two previously published studies are used, one on mobile phones and cancer and the other one on subjective symptoms.

No association between the use of cellular or cordless telephones and salivary gland tumours.

AIM: To investigate the association between the use of cellular or cordless telephones and the risk for salivary gland tumours. METHODS: Cases were assessed from the six regional cancer registries in Sweden. Four controls matched for sex and age in five year age groups were selected for each case. A total of 293 living cases and 1172 controls were included. RESULTS: There were 267 (91%) participating cases and 1053 (90%) controls. Overall no significantly increased risk was found. Odds ratios were 0.92 (95% CI 0.58 to 1.44) for use of analogue phones, 1.01 (95% CI 0.68 to 1.50) for use of digital phones, and 0.99 (95% CI 0.68 to 1.43) for use of cordless phones. Similar results were found for different salivary gland localisations. No effect of tumour induction period or latency was seen, although few subjects reported use for more than 10 years. CONCLUSIONS: No association between the use of cellular or cordless phones and salivary gland tumours was found, although this study does not permit conclusions for long term heavy use.

Case-control study on the use of cellular and cordless phones and the risk for malignant brain tumours.

PURPOSE: To investigate the use of cellular and cordless phones and the risk for malignant brain tumours. MATERIALS AND METHODS: A case-control study was performed on 649 patients aged 20-80 years of both sexes with malignant brain tumour diagnosed from 1 January 1997 to 30 June 2000. All patients were alive during the time of the study and had histopathology verified brain tumours. One matched control to each case was selected from the Swedish Population Register. The study area was the Uppsala-Orebro, Stockholm, Linköping and Göteborg medical regions of Sweden. RESULTS: Exposure was assessed by a questionnaire answered by 588 (91%) cases and 581 (90%) controls. Phone usage was defined as 'ever use' and usage starting within 1 year before diagnosis was disregarded. Overall, no significantly increased risks were found: analogue cellular phones yielded an odds ratio (OR)=1.13, 95% confidence interval (CI)=0.82-1.57, digital cellular phones OR=1.13, CI=0.86-1.48, and cordless phones OR=1.13, CI=0.85-1.50. For ipsilateral (same side) radiofrequency exposure, analogue mobile phones gave OR=1.85, CI=1.16-2.96, for all malignant brain tumours. For astrocytoma, this risk was OR=1.95, CI=1.12-3.39. For all malignant brain tumours, digital mobile phones yielded OR=1.59, CI=1.05-2.41, and cordless phones yielded OR=1.46, CI=0.96-2.23, in the analysis of ipsilateral exposure. CONCLUSION: The ipsilateral use of an analogue cellular phone yielded a significantly increased risk for malignant brain tumours.

Cellular and cordless telephones and the risk for brain tumours.

Microwave exposure from the use of cellular telephones has been discussed in recent years as a potential risk factor for brain tumours. We included in a case-control study 1617 patients aged 20-80 years of both sexes with brain tumour diagnosed between 1 January 1997 and 30 June 2000. They were alive at the study time and had histopathologically verified brain tumour. One matched control to each case was selected from the Swedish Population Register. The study area was the Uppsala-Orebro, Stockholm, Linköping and Göteborg medical regions of Sweden. Exposure was assessed by a questionnaire that was answered by 1429 (88%) cases and 1470 (91%) controls. In total, use of analogue cellular telephones gave an increased risk with an odds ratio (OR) of 1.3 (95% confidence interval (CI) 1.02-1.6). With a tumour induction period of >10 years the risk increased further: OR 1.8 (95% CI 1.1-2.9). No clear association was found for digital or cordless telephones. With regard to the anatomical area of the tumour and exposure to microwaves, the risk was increased for tumours located in the temporal area on the same side of the brain that was used during phone calls; for analogue cellular telephones the OR was 2.5 (95% CI 1.3-4.9). Use of a telephone on the opposite side of the brain was not associated with an increased risk for brain tumours. With regard to different tumour types, the highest risk was for acoustic neurinoma (OR 3.5, 95% CI 1.8-6.8) among analogue cellular telephone users.

Case-control study on concentrations of organohalogen compounds and titers of antibodies to Epstein-Barr virus antigens in the etiology of non-Hodgkin lymphoma.

A rapid increase in incidence of non-Hodgkin lymphoma (NHL) has been reported from many countries. Exposure to certain pesticides and organochlorines has been shown to be risk factors. Epstein-Barr virus (EBV) is a human herpesvirus that has been associated with some subgroups of NHL, such as Burkitt lymphoma and lymphomas related to severe immunosuppression. In this study, we measured lipid adjusted blood concentrations of 36 congeners of polychlorinated biphenyls (PCBs), p,p'-dichlorodiphenyl-dichloroethylene (p,p'-DDE), hexachlorobenzene (HCB), four different subgroups of chlordanes (trans-nonachlordane, cis-nonachlordane, MC6 and oxychlordane) and 2,2',4,4'-tetrabrominated diphenyl ether (TBDE) in incident cases of NHL and controls from the general population. Titers of antibodies to the Epstein-Barr early antigen (EA) were correlated to concentrations of organochlorines. We found a significant difference in lipid adjusted blood concentrations of total PCBs and TBDE between cases and controls. Titers of antibodies to EA IgG > 80 were correlated to an increased risk for NHL with odds ratio (OR) = 1.9, 95% confidence interval (CI) =0.94-3.8. This risk was further increased in those with a level above the median value of "sum of PCBs" (OR=4.0, CI=1.2-14), HCB (OR=5.3, CI=1.6-19), sum of chlordanes (OR=4.0, CI=1.2-14) and TBDE (OR=21, CI=4.6-124), suggesting an interaction between EBV and a higher concentration of these chemicals. Also for the "sum of immunotoxic PCBs" increased risk was found in that group (OR=6.4, CI=1.9-24). Subdivision of NHL in histological types yielded highest risks for low-grade B-cell NHL.

Adipose tissue concentrations of dioxins and dibenzofurans, titers of antibodies to Epstein-Barr virus early antigen and the risk for non-Hodgkin lymphoma.

A rapid increase in the incidence of non-Hodgkin lymphoma (NHL) has been reported in many countries. Exposure to certain pesticides or organochlorines has been shown to be a risk factor. Epstein-Barr virus (EBV) is a human herpesvirus that has been associated with some subgroups of NHL, such as Burkitt lymphoma and lymphomas related to severe immunosuppression. In this study we measured concentrations of dioxins and dibenzofurans in 33 NHL cases and 39 surgical controls. For 23 of the cases and 32 of the controls EBV titers were also available. Median titer of antibodies to EBV early antigen (EA) IgG was higher in patients than in controls. Concentrations of dioxins and dibenzofurans were divided into two groups according to the median concentration for the controls. Unconditional logistic regression analysis was performed adjusting for sex, age, and body mass index. For several higher chlorinated congeners increased risk was found for patients in the high-concentration and high-titer group. For toxic equivalency factor >27.79 and EA>80 an odds ratio of 2.8 with 95% confidence interval 0.52-18 was calculated. These results indicated that current exposure to certain organochlorines in combination with EBV might increase the risk for NHL.

Interleukin 1beta, interleukin-6, and tumor necrosis factor-alpha in endometriotic tissue and in endometrium.

OBJECTIVE: To compare the levels of IL-1beta, IL-6, and TNFalpha in endometriotic tissue and in endometrium from women with endometriosis and healthy controls. DESIGN: Open. SETTING: Department of Obstetrics and Gynecology at a university hospital. PATIENT(S): Twenty-six women with endometriosis and 22 controls operated on for clinical indications. INTERVENTION(S): ELISA in homogenized tissue samples collected during surgery. MAIN OUTCOME MEASURE(S): Levels of IL-1beta, IL-6, and TNFalpha in tissue homogenates. RESULT(S): The three types of tissue differed significantly with respect to all three cytokines. Endometriotic tissue had significantly higher concentrations of IL-1beta than endometrium from both patients with endometriosis and healthy controls. Both endometriotic tissue and endometrium from patients had significantly higher concentrations of IL-6, and endometriotic tissue had significantly lower concentration of TNFalpha than did endometrium from controls. IL-1beta showed a cycle phase dependence that was significant in endometrium from patients, being higher in the secretory than in the follicular phase. IL-1beta was significantly higher in endometrioma than in lesions of other localizations. Concentrations of IL-1beta and IL-6 were positively correlated in endometriotic tissue and in endometrium from controls. No other significant correlations were found. CONCLUSION(S): This study has shown a significant production of IL-1beta, IL-6, and TNFalpha in endometriotic tissue and endometrium, with significant differences between the tissue types, indicating a deviating cytokine pattern in both endometriotic tissue and endometrium from women with endometriosis compared with that from healthy controls.

Production of interleukins 1beta, 6 and 8 and tumor necrosis factor alpha in separated and cultured endometrial and endometriotic stromal and epithelial cells.

The production of IL-1beta, IL-6, IL-8 and TNF-alpha was studied in short-time culture of separated stromal and epithelial cells. The cytokine secretion into culture medium was analyzed using immunoassay to evaluate the cytokine protein levels and bioassay to assess the bioactivity of the cytokines. Tissue samples of endometrium and ovarian endometriomas were obtained from 4 patients operated on for clinical reasons. Only IL-8 was found in all samples. IL-1beta and TNF-alpha were detected in the culture medium from most stromal cell samples, but in fewer media from epithelial cell samples. IL-6 was measurable in a few medium samples. Few of the samples displayed a bioactivity. There was no obvious difference between endometrium and endometriotic cell samples besides the production of IL-8 that seems to be lower in endometriotic tissue.

Elevated expression of tumour necrosis factor alpha in cultured granulosa cells from women with endometriosis.

Fertilization and oocyte cleavage rates have previously been demonstrated to be lower for women with endometriosis undergoing IVF compared with controls. This might be related to impaired oocyte function, possibly due to an inflammatory milieu in the pelvis of these women, where an elevated concentration of many cytokines is documented. The aim of this study was to examine whether granulosa cells from women with endometriosis deviated with respect to production of the inflammatory cytokines interleukin-1beta, interleukin-6, interleukin-8 and tumour necrosis factor alpha (TNFalpha) compared with granulosa cells from healthy women, undergoing IVF for male infertility. The effect of human chorionic gonadotrophin on cytokine production was also investigated. Granulosa cells in follicular fluid were obtained at oocyte retrieval for IVF. Incubated cell culture media were analysed by enzyme-linked immunosorbent assay. The basal production of all four cytokines was higher in cells from women with endometriosis when compared to controls, although the increase was only significant for TNFalpha. Chorionic gonadotrophin had no significant effect, although it had a tendency to suppress cytokine release in both patient categories. Whether aberrant cytokine production in granulosa cells from women with endometriosis may disturb fertilizing capacity of oocytes requires study.

Effects of naloxone on c-jun/AP-1 in met-enkephalin-and FMRFamide-immunreactive neurons of a gastropod snail.

1. Opioid- and FMRFamide (FMRFa)-ergic systems are believed to play antagonistic behavioral roles in both higher and lower animals. In our previous experiments on a snail, behavioral choice has been demonstrated to be dependent on a balance between FMRFa and enkephalins [7]. Here, we examined if the disturbance of the balance causes changes in the activity of both systems. Opiate receptor blocker naloxone was applied and its effect on c-jun expression of met-enkephalin (MEnk)- and FMRFa-ergic neurons was examined immunocytochemically in terrestrial gastropod snail Cepaea nemoralis. 2. In control, untreated snails, central neurons with c-jun/AP-1-like-immunoreactivity were found to occur. These included MEnk-, FMRFa- and 5HT-immunoreactive (-ir) neurons, as was revealed by double-labelling. 3. After treatment with naloxone for 4 h, the following changes were observed: (i) increase in the number of MEnk-ir neurons; increase in the number of neurons showing c-jun/AP-1 and MEnk double-labeling; (ii) disappearance of c-jun/AP-1-immunoreactivity from some FMRFa-ir neurons. 4. It is suggested that immediate early genes are involved in the mechanisms responsible for the reciprocal regulation of the opioid and antiopioid neuropeptide systems.

Phosphorylation of Shc by Src family kinases is necessary for stem cell factor receptor/c-kit mediated activation of the Ras/MAP kinase pathway and c-fos induction.

In this report we show that Tyr568 and Tyr570 are phosphorylated in vivo in the Kit/stem cell factor receptor (Kit/SCFR) following ligand-stimulation. By mutation of Tyr568 and Tyr570 to phenylalanine residues and expression of the mutated receptors in porcine aortic endothelial (PAE) cells, we could demonstrate a loss of activation of members of the Src family of tyrosine kinases when Tyr568 was mutated, while mutation of Tyr570 only led to a minor decrease in activation of Src family members. Mutation of both tyrosine residues led to a complete loss of Src family kinase activation. Phosphorylation of the adapter protein Shc by growth factor receptors provides association sites for Grb2-Sos, thereby activating the Ras/MAP kinase pathway. A much lowered degree of Shc phosphorylation, Ras and Erk2 activation and c-fos induction was seen in the Y568F mutant, while in the Y570F mutant these responses were less affected. In contrast, the mitogenic response was only slightly reduced. In a mutant receptor with both Tyr568 and Tyr570 mutated to phenylalanine residues, no phosphorylation of Shc and no activation of Ras and Erk2 was seen in response to stem cell factor stimulation, very weak induction of c-fos was seen and the mitogenic response was severely depressed. These data show that Ras/MAP kinase activation and c-fos induction by Kit/SCFR are mediated by members of the Src family kinases. However, the mitogenic response is only to a minor extent dependent on Src kinase activity.

Anthraquinone-induced cell injury: acute toxicity of carminomycin, epirubicin, idarubicin and mitoxantrone in isolated cardiomyocytes.

Acute toxic effects of the antineoplastic anthraquinones carminomycin, epirubicin, idarubicin and mitoxantrone were studied in primary cultures of cardiomyocytes, which were isolated from adult rats. Both time- and concentration-dependent changes of cell structure and viability (trypan blue exclusion) following incubation of myocytes with subclinical, clinical and toxic concentrations of the anthraquinones were examined by light microscopy. The area under the decay curve of viable and rod-shaped myocytes was used to express cytotoxicity of the drugs. Mitoxantrone was found to reduce cell viability and number of rod-shaped cells to the greatest extent, followed by carminomycin, idarubicin and epirubicin. A significantly lower accumulation in cardiomyocytes was obtained with epirubicin and idarubicin compared with carminomycin. An inhibitory effect on oxygen consumption by the cells occurred already at 0.1 microM with epirubicin, whereas inhibition caused by other anthraquinones was less pronounced. Our data indicate a weak association of net accumulation and the toxicity parameter IC50 for carminomycin and idarubicin. In contrast to these results, a more significant correlation of cytotoxicity and anthraquinone lipophilicity was found, which suggests that the lipophilic character of a particular anthraquinone may be an important factor in drug-induced acute cardiotoxicity.

Identification of Tyr-703 and Tyr-936 as the primary association sites for Grb2 and Grb7 in the c-Kit/stem cell factor receptor.

In this paper we demonstrate the presence of two novel in vivo autophosphorylation sites in the c-Kit/stem cell factor receptor (c-Kit/SCFR): Tyr-703 in the kinase insert and Tyr-936 in the C-terminal tail. We furthermore demonstrate that the adapter protein Grb2 is a specific binding partner for both phosphorylated Tyr-703 and phosphorylated Tyr-936, whereas the adapter protein Grb7 binds selectively to phosphorylated Tyr-936. It is shown that the association occurs through the Src homology 2 (SH2) domains of Grb2 and Grb7. Binding of Grb2 to Tyr-703 in the c-Kit/SCFR provides a link to the Ras/mitogen-activated protein kinase pathway.

Dolichol-like lipids with stimulatory effect on DNA synthesis: substrates for protein dolichylation?

Substantial evidence has suggested that a nonsterol product of mevalonic acid (MVA) is essential for the initiation of DNA synthesis in mammalian cells. Several possible isoprenoid candidates have been suggested, but the identity of this compound still remains unknown. In this study we have isolated and purified MVA products from SV40-transformed human fibroblasts and identified fractions with a growth-stimulatory effect. The cells were labelled with [14C]MVA in the presence of inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. After lipid extraction, the [14C]MVA-labelled lipids were subjected to high performance liquid chromatography and size-exclusion chromatography, and the effect of the fractionated eluate on the DNA synthesis of arrested MVA-depleted target cells was tested. Thereby we found a fraction of [14C]MVA-labelled lipids with a substantial stimulatory effect on DNA synthesis. The chromatographic behavior suggested that the growth-stimulating fractions contained dolichol-20. This was confirmed by mass spectrometric analysis. Similar results were obtained when lipids from hepatocellular carcinoma cells and a sample from breast tumor were isolated and analyzed by the same procedure. The mechanisms by which these compounds induce DNA synthesis are unknown. Recent data obtained in our laboratory have provided evidence that dolichyl groups are covalently linked to tumor cell proteins, which implicates a new biological function for long-chain polyisoprenoid alcohols (Hjertman et al. [1997] FEBS Lett 416:235-238). In this study we demonstrate that tumor cells containing dolichol-like growth-stimulatory lipids also contained dolichylated proteins. This raises the question whether the growth-stimulatory dolichol-like lipids serve as substrates for the dolichylation reaction.

Specific and sensitive detection of the EWS/FLI1 fusion protein in Ewing's sarcoma by Western blotting.

We applied Western blotting, using an antibody against the carboxy terminal of the FLI1 protein, for detection of the 68-kDa EWS/FLI1 fusion protein in cultured Ewing's sarcoma cells and in four surgical biopsies of Ewing's sarcoma. Of six different human cell lines, the 68-kDa fusion protein was identified only in Ewing's sarcoma cells carrying the t(11;22)(q24;q12) translocation. The four samples from Ewing's sarcoma patients were also found to contain the 68-kDa fusion protein. The lowest detection level for total protein loaded on the gel was 0.3 microg. When whole Ewing's sarcoma cells were used for Western blotting without prior protein extraction, the lowest detection level was 1,300 cells. It will be possible to use Western blotting for detection of the EWS/FLI1 fusion protein in the diagnosis of Ewing's sarcoma in surgical biopsy specimens, and possibly also in fine-needle aspirates. As the method is not dependent on the quality of mRNA in the sample and involves no risk of contamination, it will be a powerful complement to the reverse-transcription polymerase chain reaction (RT-PCR).

Evidence for protein dolichylation.

Labeling of human colon carcinoma cells with [3H]dol, followed by extensive delipidation and removal of dol-P oligosaccharides, showed that dol are bound to cellular proteins with sizes of 5, 10, 27, 75 and > 140 kDa. HPLC purification of proteolytic products of [3H]dol- and [35S]cys-labeled proteins revealed a hydrophobic peak containing both dol and cysteine. The dol/cys-labeled products were clearly separated from GG-cys, and exhibited a hydrophobicity between that of dol-P and dol. In another set of experiments delipidated proteins were treated with methyl iodide, which cleaves thioether bonds. After HPLC purification of released dol-like lipids, these were subjected to mass spectrometry. This demonstrated molecular ions with the same mass as that of dol. Taken together our data provide evidence for the existence of proteins covalently modified with dol.

Mevalonate-regulated mechanisms in cell growth control: role of dolichyl phosphate in expression of the insulin-like growth factor-1 receptor (IGF-1R) in comparison to Ras prenylation and expression of c-myc.

One or more mevalonate derivatives of non-sterol type have been proposed to be of indispensable importance for cell growth. Conceivable mevalonate-dependent mechanisms involved in growth control are farnesylation of Ras proteins, regulation of c-myc expression, and N-linked glycosylation of the IGF-1 receptor. The latter mechanism might be rate-limited by dolichyl phosphate, which acts as a donor of oligosaccharides in glycoprotein synthesis in the endoplasmic reticulum. In order to study the significance for cell proliferation of the three aforementioned mevalonate-dependent processings, their inhibitory response due to mevalonate deprivation was explored and compared with the effect on DNA synthesis in the malignant melanoma cell line SK-MEL-2. We found that mevalonate depletion due to treatment with 3 microM lovastatin for 24 h, which efficiently growth-arrested the cells, hardly at all affected the expression of c-myc, and although Ras prenylation was inhibited by 50%, the most pronounced effect of lovastatin was seen on N-linked glycosylation of IGF-1 receptors, which was inhibited by more than 95%. The order and magnitude of the decreased IGF-1 receptor glycosylation, which was followed by a decreased expression of IGF-1 receptors at the cell membrane, correlated well with the inhibition of DNA synthesis. We investigated whether dolichol, and in particular dolichyl phosphate, through its participation in N-linked glycosylation, act as regulators of IGF-1 receptor expression. First, we could confirm that exogenous dolichol became phosphorylated and in this form took part in the glycosylation processing. Secondly, we showed that dolichyl phosphate, in a dose-dependent manner, could increase the number of IGF-1 receptors at the cell membrane, simultaneously as DNA synthesis was stimulated. Taken together, our results provide direct evidence for an important role of dolichyl phosphate as a regulator of cell growth through limiting N-linked glycosylation of the IGF-1 receptor.