There is general concern that persistent organic pollutants (POPs) found in the environment, wildlife, food, water, house dust, human tissues, and fluids may alter normal human physiologic activities . Although the levels of some POPs [polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCs)] in these matrices have decreased after their ban, others [polybrominated diphenyl ethers (PBDEs)] have increased in recent years.To determine the longitudinal trend of specific POPs in human fetal tissues for risk assessment purposes.n = 52) and placental (n = 60) tissues, obtained after elective abortions during 1998–2006, for selected PBDEs, PCBs, and OCs using gas chromatography–mass spectroscopy.We analyzed early to mid-gestation fetal liver (p < 0.03), whereas placental levels were generally lower, with no clear trend. Low levels of PCBs and OCs varied yearly, with no evident trend. The major analytes in 1998 were OCs , whereas the major analytes in 2006 were PBDEs . The 1998–2006 tissue PBDE congener profile is similar to that of DE-71, a commercial primarily pentabrominated diphenyl ether mixture manufactured in North America.Total PBDEs in fetal liver increased over time (mean ± SE: 1998, 284.4 ± 229.8 ng/g lipid; 2006, 1,607.7 ± 605.9; Although commercial production of penta- and octa-brominated diphenyl ethers in North America was halted in 2004, their concentrations in fetal liver and placenta are now greater than the tissue burdens for the analyzed OCs and PCBs. Our findings also demonstrate that PBDEs accumulate within the fetal compartment at a very early stage in gestation. Persistent organic pollutants (POPs), such as organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs), contaminate many environmental matrices and can be found in human blood, adipose tissue, and breast milk . In addiin utero exposure and fetal outcome. A more recently recognized class of environmental contaminant, whose chemical structure is similar to PCBs, is the brominated flame retardants, which includes the polybrominated diphenyl ether (PBDE) congeners . For theIn contrast to OCs and PCBs, many studies worldwide have reported that the amount of PBDEs found in the environment, human serum, and breast milk has been increasing steadily since the 1980s . A few iin utero exposure begins to contribute significantly to an infant’s body burden of PBDEs and other POPs. To examine this question, we analyzed the levels of multiple OCs p,p′-DDT , p,p′-DDE , trans-nonachlor, and hexachlorobenzene (HCB)], PCB congeners , and BDE congeners  in early to mid-gestation fetal liver and placental specimens obtained between 1998 and 2006. The results are an expanded version of data presented previously at two scientific meetings  samples  after elective pregnancy terminations during 1998–2006 in the Greater Montreal area of Quebec with informed written consent. Fetal ages were determined by foot length . The tisWe precleaned all glassware, except volumetric flasks and custom-made columns, in a dishwasher and baked them at 475°C for 2 hr in a Tempyrox oven . The volumetrics and custom-made columns were cleaned by solvent rinsing with acetone followed by hexane. We thawed approximately 1 g of each sample at room temperature in 50-mL centrifuge tubes and homogenized them in 20 mL of a 2:1 acetone/hexane solution for 1 min using a Polytron homogenizer . The resulting supernatant was quantitatively transferred and filtered through precleaned (dichloromethane) glass wool, collected in 250-mL round-bottom flasks (RBFs), and concentrated to approximately 1 mL on a rotary evaporator . The extracts were dried by pouring them into funnels containing prebaked sodium sulfate  and again collected in 250-mL RBFs. We then transferred the dried extracts to preweighed 10-mL vials for gravimetric lipid determinations.p,p′-DDT, p,p′-DDE, trans-nonachlor, HCB). We selected these analytes based on previous studies of Canadian breast milk samples showing that they had the highest levels and occurrences in 1992 and 2002 , PCB congeners , and several OCs (and 2002 . In addiand 2002 . The Flohttp://www.ehponline.org/members/2008/0800205/suppl.pdf) provides the mass-to-charge ratios monitored for the target ions and qualifier ions as well as the expected qualifier response factors and response factor tolerances.We analyzed samples using an Agilent 6890N/5973 GC-MS system operated in electron impact ionization mode equipped with an Agilent 7683 autosampler . Aliquots of 2 μl were injected via an on-column injector operated in oven-track mode. The capillary column employed was a 30 m × 0.25 mm × 250 μm DB5-MS (Agilent Technologies) coupled to a 2-m × 0.53-mm deactivated fused silica column . The GC oven parameters were 80°C for 2 min; 8°C/min to 220°C, 4 min hold; 5°C/min to 230°C; 10°C/min to 295°C; 10 min hold. The helium carrier gas was set at a constant flow rate of 1.2 mL/min and the transfer line temperature was maintained at 295°C . Supplemental Material, http://www.ehponline.org/members/2008/0800205/suppl.pdf). Analyte concentrations in the samples were calculated based on the equation of the linear regression line after forcing through the origin. The response was linear for all analytes (r2 > 0.99).External standard calibration was used after determination of the instrument’s linear response range for each analyte. We based positive identification of target compounds on three criteria: a chromatographic retention time matched to analytical standards, a signal-to-noise ratio of > 3, and qualifier ion ratios within the tolerances listed in Supplemental Material, http://www.ehponline.org/members/2008/0800205/suppl.pdf)]. The BDE background contamination levels fluctuate almost daily, as determined from the reagent blanks; therefore, we subtracted this value from the spike, the SRM, and the tissue samples on a batch-to-batch basis. Control charts were used to monitor the performance of the SRM and corrective action was undertaken any time the warning limit (mean ± 2σ) was approached. Recoveries from the spiked corn oil sample were also monitored on a batch-to-batch basis. Supplemental Material, Table 2, provides mean recovery values (http://www.ehponline.org/members/2008/0800205/suppl.pdf). Method detection limits (MDLs) for each analyte were determined using eight corn oil samples (100 mg) spiked with a known concentration of each analyte; linearity was established between 7 and 900 ng/mL . The corn oil spikes were then extracted as outlined above and the mean concentrations for each analyte calculated. We calculated the MDL as the SD for the replicates (n = 8) multiplied by 2.31 . Supplemental Material, Table 2, lists MDLs (http://www.ehponline.org/members/2008/0800205/suppl.pdf).Instrument sequences consisted of one reagent blank, one corn oil sample (no background POPs) spiked with the analytes, one standard reference material (SRM) sample , and seven tissue samples  of total PBDEs, PCBs, and OCs. 10 tissue concentration, nanograms per gram lipid); the dashed line represents the trend with time (1998–2006) using all of the samples per year , and the solid line, the trend using only samples with detectable levels of PBDEs. We analyzed the data using a nonparametric approach because of the existence of zeros (nondetects) and values below the MDL that prevented a transformation from achieving normality. The nonparametric measure of correlation, Kendall’s tau statistic ]. Of the PCBs, only PCB 180 showed a significant trend: a decrease with time and only in liver . The major PCB congener in both tissues was PCB 153 .Kendall’s tau test for trend revealed no significant changes from 1998 through 2006 in any of the individual OC analytes or total OCs in either the fetal liver or placental tissues. The major OC in both tissues was r = 0.22, p < 0.03; r = 0.20, p < 0.04; r = 0.24, p < 0.02; r = 0.29, p < 0.006; data not shown but similar to BDE-154), BDE-154 . In the placental specimens, there was a significant increase in BDE-85 . Because of the limited number of samples per year, it was not possible to analyze statistically whether there was a correlation between the age of the fetus and the amount of POPs in the tissue samples; however, we found no apparent associations when we graphed the data (data not shown).In utero exposure is also considered to be a critical period  in the 1970s and 1980s has produced a decline in their relative concentrations in various matrices, including human specimens, during the past 30 years . In addiSince the 1990s, studies of environmental and human specimens in North America have uniformly documented a dramatic increase in PBDE levels . What isOn average, the concentrations of PBDEs as well as OCs and PCBs in our liver samples were appreciably higher than those in the placenta, suggesting tissue-specific bioaccumulation. This may be due in part to the relative difference in their lipid content . However, the lack of a correlation within the paired samples in our study, as well as the results from investigations of paired maternal and cord sera , suggestAlthough 84% of our PBDE values were in the range of 0–1,500 ng/g lipid, two liver and three placental samples had levels approximately 10- to 100-fold higher. Unfortunately, these were not among the paired samples, but that may not have provided much insight into whether the mother’s BDE burden greatly exceeded the population’s average because we found no association between the amount of PBDEs in our fetal liver and placental samples. We analyzed samples collected anonymously, therefore neither our present study nor that of Interestingly, ours is not the only report of PBDE concentrations that vary markedly among samples. In a review of Canadian breast milk studies, In summary, our study of POPs in human fetal liver and placental tissues suggests that PBDEs, PCBs, and OCs are passing into and accumulating within the fetal compartment from a very early stage in gestation. Although we found little change in the relative amounts of the PCBs and OCs from 1998 through 2006, there has been a significant increase in the fetal liver levels of PBDEs. These data, along with the results from previous studies of human postnatal blood, breast milk, and adipose tissues, suggest increased exposure of humans to PBDEs at every stage in life during the past decade. Although this appears to be more of an issue in North America, the widespread presence of PBDEs in commercial and household items as well as in the environment and food chain ensures that this is a universal problem. Whether some individuals are reaching an exposure level that may constitute a danger to their health is unknown. Annual monitoring programs  in different regions would help to answer how the body burdens are changing in the future and provide important dose information for studies of the physiologic effects of these contaminants.


The relationship between prostate cancer and height is uncertain.We prospectively examined the association of height with prostate cancer among 34268 men in the prostate, lung, colorectal, and ovarian cancer trial. Anthropometry was assessed at baseline and 2144 incident prostate cancer cases were identified upto 8.9 years of follow-up.P trend=0.05; relative risk (RR) for 190 cm+ vs ⩽170 cm=1.39, 95% confidence interval (95% CI): 0.96–2.01). This association was largely limited to men below the age of 65 years , although the number of cases was small and risk estimates were somewhat unstable.Overall, tallness was not associated with the risk of prostate cancer or with the risk of non-aggressive disease, but the risk for aggressive prostate cancer tended to be greater in taller men (Gleason score ⩾7 or stage ⩾III; The results of this large prospective prostate cancer screening trial suggest that tallness is associated with increased risk for younger onset aggressive prostate cancer. Tallness is a potential risk factor for prostate cancer , possiblThe PLCO Screening Trial is a multi-centre trial designed to evaluate screening methods for the early detection of these cancers (n=775); men without an initial PSA test or DRE (n=2470); men who received an initial screening examination, but with whom there was no subsequent contact (n=721); men who did not complete the baseline questionnaire (n=898); and men with missing (n=205) or extreme values for height . We also excluded men whose initial screening examination occurred after 30 September 2002, the censoring date for this analysis (n=72). After exclusions, the analytic cohort comprised 34 268 men (some participants were included in more than one exclusion category).Of the 38 349 men who were randomly assigned to the screening arm of the trial, we excluded men who reported a history of cancer other than non-melanoma skin cancer  and had a Gleason sum of <7 (low-grade disease). Tumours were considered aggressive if classified as stage III or IV (high-stage disease) or assigned a Gleason sum of ⩾7 (high grade-disease). We also distinguished between high-grade disease (defined as a Gleason sum of ⩾7) and high-stage disease (defined as stage III or IV).2 or more), and number of screenings using PSA or DRE during the trial. Tests for linear trend were conducted by assigning the median value for each category and treating that term as a single continuous variable in the model. In addition, stratified analyses were carried out on the basis of age at baseline, history of prostate cancer, race, history of diabetes, current BMI, and BMI at age 18 years. We formally tested for interactions using log-likelihood ratio tests. All the analyses were conducted using SAS version 9.1 . All P-values were two sided.Person-years were calculated from the date of the baseline prostate cancer screening to the date of the most recently completed annual follow-up questionnaire, date of diagnosis, death, or 30 September 2002, whichever came first. We used Cox proportional hazards regression analysis to estimate RRs and 95% CIs. Multivariable analyses were adjusted for confounding factors, including age (continuous), race , PLCO study centre, family history of prostate cancer , smoking status , education , history of diabetes , body mass index (BMI) . Positive trends with height were noted for both high-grade  and high-stage disease .During 170 882 person-years of follow-up of 34 268 men, 2144 men were diagnosed with prostate cancer, of whom 1202 (57%) were classified as non-aggressive and 912 (43%) as aggressive cases. In multivariable analysis, height was associated neither with the risk of prostate cancer overall nor with non-aggressive disease . HoweverP trend=0.008; RR=1.76, 95% CI=1.06–2.93; comparing 190+ with ⩽170 cm). No clear pattern was observed for men in the older (⩾65 years) age group. The test for interaction between age and height was statistically significant (P interaction=0.009). There was no interaction between height and other risk factors  in relation to risk .As exploratory analyses, we have examined whether relationships with height differ according to prostate cancer risk factors. A strong positive association with aggressive disease was observed for men younger than 65 years   (n=109), prevented our examining potential interaction with family history. Our findings require confirmation in large consortium-based studies.Our finding of a stronger association with younger onset aggressive prostate cancer is consistent with a prospective study in Hawaii (<72.5 years, Height is influenced by bioavailable IGFs and androgens . InsulinA limitation of our study is that height was assessed by self-report and hence was not validated. Nevertheless, the correlations between measured and self-reported height are typically high, ranging from 0.80 to 0.95 , althougThis large prospective study suggests that tallness may be associated with increased risk of aggressive prostate cancer, particularly in younger men; confirmation is needed in other populations.


Objectives. Mitral valve prolapse (MVP) is a known cause for false positive exercise test (ET). The purpose of this study was to establish additional electrocardiographic criteria to distinguish the false positive exercise results in patients with MVP. Methods. We studied 218 consecutive patients  with MVP (according to echocardiographic study), and positive treadmill ET was performed due to multiple cardiovascular risk factors or angina-like symptoms. A coronary angiography was performed to detect coronary artery disease (CAD). Results. From 218 patients, 90 (group A) presented with normal coronary arteries according to the angiography  while the rest 128 (group B) presented with CAD. ST-segment depression in hyperventilation phase was present in 54 patients of group A (60%) while only in 14 patients of group B (11%), P < .05. Conclusions. Presence of ST-segment depression in hyperventilation phase favors a false positive ET in patients with MVP. Mitral valve prolapse (MVP) is one of the most prevalent cardiac valvular abnormalities. Using standardized echocardiographic diagnostic criteria, a community-based study showed that MVP occurs in 2.4 percent of the population , 2. It i The vast majority of patients with MVP are asymptomatic and remain so throughout their lives. However, some patients may complain of symptoms like syncope, pre-syncope, palpitations, chest discomfort, and, when mitral regurgitation (MR) is severe, symptoms of diminished cardiac reserve. Chest discomfort may be typical of angina pectoris but is more often atypical in that it is prolonged, not clearly related to exertion, and punctuated by brief attacks or severe stabbing pain at the apex. In these patients false positive exercise stress responses, as testified by coronary angiography, electrocardiographic –14 as we Since the diagnosis of coronary artery disease (CAD) in patients with MVP based on the standard ST-segment depression criteria during exercise testing (ET) is uncertain, the establishment of accessional electrocardiographic (ECG) criteria for the discrimination of true positive exercise responses would be of great value.  It is well known that patients with MVP often have high levels of catecholamines –22. On tFrom 2006 to 2009, 218 consecutive patients with MVP  and a normal resting ECG who presented with ischemic ECG changes during ET and agreed to undergo coronary angiography were included in the study. During the recruitment period, a total of 746 patients with MVP were examined. From them, 252 (34%) presented with ischemic ECG changes during ET in our laboratory, and 218 (87%) accepted to participate in our study. The diagnosis of MVP was based on an echocardiographic study while the definition of ischemic ECG response during ET was based on the standard ST-segment deviation criteria. Treadmill ET was performed in these patients due to multiple cardiovascular risk factors or angina-like symptoms. Patients with known history of CAD, left or right bundle-branch block, left or right ventricular hypertrophy, other valvular abnormality, pre-excitation syndromes, and those who were receiving digitalis or were unable to walk on treadmill were excluded from the study. All medications were discontinued for at least 5 half-lives before the ET. The study was approved by our hospital's committee on human research, and written informed consent was obtained from all participants.Echocardiographic study was performed to all patients in order to establish the diagnosis of MVP . The two From 218 patients with documented prolapse, 180 (83%) had typical auscultatory findings (click or murmur). Doppler echocardiography frequently revealed mild MR that was not always associated with an audible murmur. Moderate to severe MR was present in about two-thirds of patients with posterior leaflet prolapse and in about one-fourth of patients with anterior leaflet prolapse which is in concordance with the literature .All patients performed ET on a Marquette case system , according to the multistage Bruce protocol. Each patient had an electrocardiogram (ECG) recorded with the standard 12 leads. The ECG was recorded continuously at the resting status (both supine and standing), during hyperventilation, during exercise, and for up to the 10th minute of the recovery period. Blood pressure was measured every minute at the resting status (both supine and standing), during hyperventilation, during exercise, and during the recovery period, with a sphygmomanometer. Exercise was terminated in the presence of exercise-limiting angina, fatigue, dyspnea, ataxia, dizziness, near syncope, or signs of poor perfusion . In the absence of symptoms, the test was terminated at the occurrence of ≥ 0.2 mV ST-segment depression or ≥ 0.1 mV ST-segment elevation, an exaggerated blood pressure response (≥ 250/115 mm Hg), a decrease in systolic blood pressure ≥ 10 mm Hg from baseline or specific arrhythmias  .2: 21%) with a frequency rate of 40–45 breaths/min at room temperature (20–22°C). The target ventilation rate was equivalent to the maximum voluntary ventilation, and the mean duration of the test was 1 minute. Hyperventilation was discontinued earlier if the patient felt dizziness, lightheadedness, weakness, shortness of breath, a sense of unsteadiness, tingling around the mouth, fingertips, and toes (as a result of hypocapnia) in order to avoid loss of consciousness. All patients underwent forced hyperventilation (breaths as deep and rapid as possible—at least 30 breaths/min) in the standing position before the exercise. The test required the subject to overbreadth voluntarily the room air  as horizontal or downsloping depression of ≥ 0.1 mV below the resting ST-segment level (measured 60 milliseconds after the J-point), upsloping depression of ≥ 0.15 mV (measured 80 milliseconds after J-point), or elevation of ≥ 0.1 mV . Intensive care was taken to detect ST-segment depression in any of the leads during the hyperventilation phase of the test since all patients presented with normal baseline (both supine and standing) ECG. In the presence of ST-segment depression during hyperventilation, an ischemic ST-segment response during exercise was defined as additional depression of ≥ 0.1 mV (measured in the standing position) below the hyperventilation ST-segment level . Exercise tests without ischemic ST-segment changes, which were terminated when the heart rate was < 85% of the predicted maximal heart rate, were considered inconclusive. Patients with inconclusive exercise tests were excluded from the study. ECG measurements were performed with a magnifying lens by two of the investigators who were unaware of the results of the coronary angiography. The interobserver and intraobserver variability for ST-segment changes was 0.007 ± 0.005 and 0.008 ± 0.004 mV, respectively. Disagreements between observers were resolved by a third interpreter.Selective coronary arteriography was performed to all patients, using the percutaneous (Judkins) technique. The left coronary artery was visualized at the 60° left anterior oblique, the 30° right anterior oblique, and the left lateral position, with a 30° cranial angulation. The right coronary artery was visualized at the 60° left anterior oblique and the 30° right anterior oblique position. Significant coronary stenosis was defined as diameter narrowing of 70% or more in the lumen of left anterior descending (LAD), left circumflex (LCx), and right coronary artery (RCA), or 50% or more in the lumen of left main coronary artery. The interpretation was performed by two investigators who were unaware of ET results.t-test. Categorical variables between two groups were compared using chi-square test (with the use of Yates correction) or Fisher's exact t-test. All tests were two-sided. Differences were considered as statistically significant for P  value < .05. Data analysis was performed with SPSS 15.0 statistical software .All continuous variables were normally distributed as confirmed by using Kolmogorov-Smirnov test and are expressed as mean ± SD. Categorical variables are reported as observed number (percentage). Comparisons of continuous variables between two groups were performed by using unpaired From 218 patients included in the study, 103 (47%) were males while the rest 115 (53%) patients were females. Patient's characteristics, echocardiographic and angiographic data are demonstrated in  There were no differences between males/females regarding the mean age of the patients or the prevalence of arterial hypertension, diabetes mellitus, hypercholesterolemia, and smoking.P = .048 while posterior leaflet prolapse was present in 23 males (22%) and 38 females (33%), P = .042. The ejection fraction of the left ventricle was similar in men and women. Anterior leaflet prolapse was present in 80 males (78%) and 77 females (67%), P = NS for all. Overall, from 218 patients with positive ET, 90 (41%) had normal coronary angiograms . Coronary vasospasm was not observed in any patient during angiography. Normal coronary arteries were found in 38 males (37%) and 52 females (45%) while 1-vessel disease was found in 33 males (32%) and 37 females (32%), 2-vessel disease in 21 males (20%) and 18 females (16%), and 3-vessel disease in 11 males (11%) and 8 females (7%),  Exercise and ECG data are presented in P = .036). Hyperventilation induced an increase in systolic blood pressure  and heart rate . Exercise parameters  were similar in males and females with the exception of the duration (in sec) of the exercise  presented with ST-segment depression in the inferior leads , 2 patients (3%) in the anterior leads , 12 patients (18%) in the lateral leads  while the rest 23 patients (34%) in both inferior and lateral leads. Furthermore, 27 patients (40%) presented with horizontal and the rest 41 patients (60%) with downsloping pattern of ST-segment depression. This depression was measured (in mV) up to 0.09 ± 0.03 in men versus 0.11 ± 0.05 in women, P = NS . ST-segment depression during the hyperventilation phase of the test was detected in 27 males (26%) and 41 females (36%), P = NS . Maximal exercise-induced ST-segment depression (in mV) was measured at peak exercise up to 0.15 ± 0.02 in men versus 0.19 ± 0.03 in women, P = .043). Time required for exercise-induced ST-segment depression of 0.1 mV (in sec) was greater in males . Overall, from 68 patients with ST-segment depression during hyperventilation, 54 (79%) appeared without CAD  which is concordant with the literature [ The results of this study indicate that false positive exercise response is common in patients with MVP ≈ 1% which terature . Hyperventilation-induced ST-segment depression is a criterion that (according to the results of this study) makes CAD less likely in MVP patients with exercise-induced ischaemic response. The incorporation of this new electrocardiographic criterion in the standard ST-segment depression criteria makes exercise stress testing much more accurate in detecting CAD in MVP patients. With sensitivity up to 89% and reduced false positive responses compared to standard ET (from 41% to 24%), a normal ECG during hyperventilation makes an ischaemic stress response in patients with MVP quite trustworthy. No previous data are available concerning the ability of ST-segment changes during hyperventilation to detect coronary artery disease in patients with MVP when accompanying with positive exercise-induced responses. Ellestad et al. report that patients with abnormal autonomic drive (high levels of catecholamines) have demonstrated ST-segment depression after hyperventilation as well after exercise . In thei2+ overload, and oxidative stress play essential role in the pathogenesis of catecholamine-induced myocardial damage [2+-handling abnormalities in both sarcolemmal and sarcoplasmic reticulum, defects in energy production by mitochondria, and myocardial cell damage [ Mechanisms involving disturbed balance between oxygen demand and supply, decreased level of high-energy phosphate stores, changes in electrolyte contents, intracellular Cal damage . Severall damage . Hypervel damage –22, 33 al damage .  Considering all these potential pathophysiological mechanisms and that patients with high levels of catecholamines frequently present with ST-segment depression during hyperventilation in the absence of CAD –27, the Absence of ST-segment changes during hyperventilation in MVP patients with positive exercise testing is proved to be a useful diagnostic tool for detecting CAD with Se, Sp, Ppv, Npv, and Ac up to 89, 60, 76, 79, and 77%, respectively. Reducing false positive responses (from 41% to 24%), this novel criterion could guide physicians to avert unnecessary coronary arteriography in an amount of patients with MVP . Since exercise ECG test is a popular, well-established, inexpensive, widely available procedure, we should probably consider it as an acceptable diagnostic tool for CAD even in patients with MVP after incorporating this novel ECG criterion.The results of our study would be more explicable if catecholamines levels had been measured in our patients. Besides, it should be clearly stated that unless an MVP diagnosis is in fact known, the study results do not apply to other patient groups.


Female and male RTs reported a total of 111 and 34 haematopoietic malignancies and 115 and 34 solid tumours, respectively, in their offspring. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated using Cox proportional hazards regression. Leukaemia (n=63) and solid tumours (n=115) in offspring were not associated with maternal in utero or preconception radiation exposure. Risks for lymphoma (n=44) in those with estimated doses of <0.2, 0.2–1.0, and >1.0 mGy vs no exposure were non-significantly elevated with HRs of 2.3, 1.8, and 2.7. Paternal preconception exposure to estimated cumulative doses above the 95th percentile  was associated with a non-significant risk of childhood cancer of 1.8 (95% CI 0.7–4.6). In conclusion, we found no convincing evidence of an increased risk of childhood cancer in the offspring of RTs in association with parental occupational radiation exposure.We examined the risk of childhood cancer (<20 years) among 105 950 offspring born in 1921–1984 to US radiologic technologist (USRT) cohort members. Parental occupational  As 73% USRT cohort participants are female, both preconception and n=23 123; the latest year for which estimated doses were available); had missing, incomplete, or inconsistent data on birth year, parent birth year, cancer diagnosis year, or death year (n=2859); had reported Down's syndrome (n=123); or had an ‘other cancer’ reported as a condition of the skin (other than sarcoma) or the cervix (n=73) occurring during childhood (<20 years), to eliminate potentially benign common conditions of these tissues . The resultant data set included 81 262 and 24 678 offspring of female and male RTs, respectively.The University of Minnesota Institutional Review Board and the United States National Cancer Institute approved all protocols for data use. Complete study details have been described elsewhere  and 95% confidence intervals (CIs) were calculated by Cox proportional hazards regression . Person-time was calculated as the interval between the reported birth year and the cancer diagnosis year, death year, the year the child turned age 19, or the year that the second questionnaire was completed, whichever came first. A person-time of 0.5 years was assigned to those who were censored or had an event in their birth year. All regression analyses were stratified by the gender of the parent.Average cancer incidence rates per million person-years were determined for each age group in years  as the number of events per person-years. The ratio of observed to expected numbers (the standardised incidence ratio (SIR)) was calculated for RT parent and child gender strata for childhood leukaemia, lymphoma, and all sites by applying the sex-specific incidence rates obtained from two registries: (1) Surveillance, Epidemiology, and End Results (SEER) 9 (1973–2004)  and (2) P-values for tests for linear trend calculated by inclusion of the continuous measure of dose in regression models. Estimated in utero dose exposures were categorized as follows: 0 mGy (reference), >0–0.17 mGy, 0.18–1.0 mGy, and 0.01–12.6 mGy. Estimated preconception ovarian and testes dose exposures were categorized according to quartiles: <0.43 mGy, 0.43–1.49 mGy, 1.50–3.57 mGy, >3.57 mGy; testes: <0.67, 0.67–4.92, 4.93–15.26, >15.27) with additional analyses for the offspring of males that divided the fourth quartile at the 95th percentile (>81.92 mGy).Exposures were modelled as categorical variables with n=68), results are given for haematological malignancies (leukaemia and lymphoma), solid tumours, and overall childhood cancers only.For the offspring of female RTs, results are reported for the following childhood cancer outcomes: (1) leukaemia, (2) lymphoma, (3) solid tumours, and (5) all childhood cancers. Owing to the small number of childhood cancer cases reported by male RTs . Overall, case parents tended to be younger (<30 years) than non-case parents at the time of the offspring's birth (79 vs 73%) (data not shown).Approximately 96% of cases and 95% of non-cases had parents who reported being white. Cases and non-cases were similar with respect to birth order (48% of cases n=94), lymphoma (n=61), and central nervous system (CNS) tumours (n=46) were the most common cancer types. The peak leukaemia incidence occurred at <5 years of age with rates of 72 and 97 cases per million person-years in female and male RT offspring, respectively. Peak lymphoma incidence occurred in older children, with the highest rate being in 15- to 19-year olds. Central nervous system tumour rates did not show marked variation by age in the offspring of female RTs. The pattern for the offspring of male RTs was less consistent, presumably due to the small number of cases (n=12).In total, 294 offspring had reported cancers diagnosed at <20 years of age . Leukaemin utero radiation exposure (P=0.32).No significant increase in risk or dose–response was found for leukaemia, lymphoma, solid tumours, or childhood cancer overall in association with exposure . Based oNo appreciable increased risk or dose–response was observed between maternal preconception exposure and childhood cancer. The estimated paternal doses above the 95th percentile (>82 mGy) were associated with a non-significant increased risk of 1.8 (95% CI 0.7–4.6) relative to the reference group, on the basis of six events  .Standardised incidence ratios were calculated for leukaemia, lymphoma, or all reported childhood cancer rates in the offspring of RTs relative to SEER and Connecticut registry rates . Cancer in utero or before conception. An increased risk of lymphoma was observed based on a small number of cases but there was no dose–response. Our data do not indicate an association between maternal preconception radiation exposure and offspring childhood cancers. A non-significant increased risk of childhood cancer was associated with paternal preconception exposure only above the 95th percentile (>82 mGy), based on few cases.Overall, our results do not support an increased risk of childhood cancer in the offspring of RTs associated with occupational IR exposure while In utero exposure to maternal occupational medical radiation did not markedly increase the risk of any of the childhood cancer outcomes examined except lymphoma. A small increase of approximately 40% in the risk of childhood cancer is thought to be detectable for acute exposure to diagnostic in utero radiation at doses in the range of 10 mGy, based on results from case–control studies of subjects born between late 1940s and early 1980s (in utero dose for lymphoma cases was 0.3 mGy, which is approximately 10-fold lower than average annual background radiation exposure (ly 1980s . The offexposure  and, theConsistent with our results, most previous studies have not provided strong support for an association between childhood cancer and parental preconception exposures to either low or high doses of atomic bomb radiation , medicalin utero diagnostic radiation exposure in two of the largest studies: the Oxford Survey of Childhood Cancers (in utero (Excess in cases of lymphoma in the male offspring of those exposed to radiation has not been previously reported. No male excess has been reported in association with  Cancers  and the  Cancers , or in tOur study addresses the important issue of cancer risk in the offspring of medical radiation workers, particularly females, who are exposed to low-level protracted occupational radiation. Prior data comes mainly from studies of cancer risk in offspring of nuclear workers who are predominantly male. However, as with all studies of rare diseases and low-dose exposures, our study has its own limitations. Electronic files of film-badge doses were not routinely available until late 1970s. To estimate these doses, we undertook a comprehensive dose reconstruction that used hundreds of thousands of badge doses from electronic files from 1977 onward, thousands of badge doses from hard copy records for the period 1960–76, and literature-based dose data for the period before 1960 (Although our sample size is insufficient to detect small increases in risk, we can conclude with certainty that the risk of cancer is not greatly increased in offspring of radiologic technologists.


Toxocara canis can invade the brain and cause BI in humans and mice, leading to cerebral toxocariasis (CT). Because the parasitic burden is light in CT, it may be too cryptic to be detected in humans, making it difficult to clearly understand the pathogenesis of subtle BI in CT. Since the pathogenesis of murine toxocariasis is very similar to that in humans, it appears appropriate to use a murine model to investigate the pathogenesis of CT.Because the outcomes and sequelae after different types of brain injury (BI) are variable and difficult to predict, investigations on whether enhanced expressions of BI-associated biomarkers (BIABs), including transforming growth factor β1 (TGF-β1), S100B, glial fibrillary acidic protein (GFAP), neurofilament light chain (NF-L), tissue transglutaminases (tTGs), β-amyloid precursor proteins (AβPP), and tau are present as well as whether impairment of the ubiquitin-proteasome system (UPS) is present have been widely used to help delineate pathophysiological mechanisms in various BIs. Larvae of T. canis embryonated eggs was investigated from 3 days (dpi) to 8 weeks post-infection (wpi) by Western blotting and RT-PCR.BIAB expressions and UPS function in the brains of mice inoculated with a single dose of 250 T. canis larvae were found to have invaded areas around the choroid plexus but without eliciting leukocyte infiltration in brains of infected mice; nevertheless, astrogliosis, an indicator of BI, with 78.9~142.0-fold increases in GFAP expression was present. Meanwhile, markedly increased levels of other BIAB proteins including TGF-β1, S100B, NF-L, tTG, AβPP, and tau, with increases ranging 2.0~12.0-fold were found, although their corresponding mRNA expressions were not found to be present at 8 wpi. Concomitantly, UPS impairment was evidenced by the overexpression of conjugated ubiquitin and ubiquitin in the brain.Results revealed that at 4 and 8 wpi, Further studies are needed to determine whether there is an increased risk of CT progression into neurodegenerative disease because neurodegeneration-associated AβPP and phosphorylated tau emerged in the brain. Brain injury (BI) caused by any number of physical, chemical, or biological insults can have disabling or even fatal consequences . BecauseTGF-β1 is involved in regulating the brain's response to inflammation and injury. Increased levels of TGF-β1 have been correlated with deposition of scar materials after traumatic CNS injury, and overexpression of TGF-β1 in the CNS might lead to reduced microglial activation and reduced induction of proinflammatory chemokines after severe hypoxic-ischemic injury . The S10Astrocytes are supportive structural elements of the nervous system. They play active roles in normal brain physiology and in certain pathological conditions. Reactive astrocytes with increased expression of GFAP are commonly found in cerebral infarction and many areas of brain damage . NF-L is2+-dependent enzyme, can catalyze the incorporation of a polyamine into polypeptide-bound glutamine leading to the formation of a (γ-glutamyl) polyamine bond resistant to proteolytic cleavage, and may serve not only to stabilize proteins against degradation, but may also alter their functions. Elevated expression of tTG has been observed after traumatic BI (TBI) [tTG, a CaBI (TBI)  and cereBI (TBI) . AβPPs, BI (TBI) , and wheBI (TBI) .The normal function of the tau protein, an important structural element in neuronal cells, is to assemble and stabilize microtubules, but in certain BIs , it is redistributed to cell bodies where it accumulates and forms insoluble, fibrillary deposits . The ubiToxocara canis is an intestinal nematode of canines, and its embryonated eggs are infectious to both final and paratenic hosts including humans and rodents. Toxocara canis larvae, measuring 357~414 μm in length × 13~17 μm in width [T. canis larvae and migratory pathways [in width . Howeverin width -21. Sincpathways , it may T. canis larvae by measuring levels of BI-associated indicator molecules of TGF-β1, S100B, GFAP, NF-L, tTG, AβPP, and tau as well as UPS function by assessing ubiquitin expression in infected mice brains.In this study, we investigated the pathogenesis of CT and the extent of neural damage caused by Toxocara canis eggs were obtained from adult female worms, and embryonated eggs were prepared as previously described [ad libitum. The viability of T. canis embryonated eggs was assessed by the light stimulation method immediately before use. Each mouse was infected by oral intubation with about 250 embryonated eggs in 100 μl of water [escribed . Female of water .Policies and Guidelines for the Care and Use of Laboratory Animals, Taipei Medical University and all effort was made to minimize animal suffering.Infected mice were deeply anaesthetized with ether and killed by heart puncture on day 3 and at weeks 1, 4, and 8 post-infection (dpi or wpi). On each date, three infected mice and two age-matched uninfected mice were killed, and their brains were excised for further experimentation. As shown in Figure g for 10 min), and the larvae in the sediment were counted in a Petri dish placed under an inverted microscope  at 100× magnification.Whether the larvae had successfully invaded the brain was confirmed using a previously described method . BrieflyA brain specimen from each mouse was separately fixed in 10% neutral buffered formalin for at least 24 h and embedded in paraffin for pathological studies and immunohistochemical detection of GFAP. Five-micrometer brain sections were processed using standard procedures and stained with hematoxylin-eosin (H&E) for the histological study. For the immunohistochemical studies, brain sections from infected and uninfected mice were deparaffinized and rehydrated using descending ethanol gradients before further processing. Immunohistochemical detection of GFAP was performed as described by Balasingam et al. [g at 4°C for 10 min, and stored at -80°C for further studies. Before being loaded onto the gel, the protein extracts were boiled for 5 min. In each lane, 50 μg of proteins in loading buffer was boiled and then electrophoresed in a 6%~18% SDS/PAGE mini gel. The proteins were then electrically transferred onto a Hybond-P polyvinylidene fluoride membrane for 2 h. Membranes were blocked in 10% PBS/skimmed milk, and then mouse anti-TGF-β1 monoclonal antibodies (mAbs) , mouse anti-S100B mAbs , mouse anti-tau mAbs , mouse anti-β-actin mAbs , goat anti-tTG polyclonal antibodies (pAbs) , goat anti-NF-L pAbs , rabbit anti-GFAP pAbs , mouse anti-AβPPs mAbs , mouse anti-Aβ mAbs , or mouse anti-ubiquitin mAbs  were added for 2 h at 37°C. After several washes with PBS-Tween 20, a peroxidase-conjugated secondary antibody was added, and the membrane was hybridized at 37°C for 30 min. Immunoreactions were detected with a Western Lightning® kit . The secondary antibodies, rabbit anti-mouse immunoglobulin G (IgG), goat anti-rabbit IgG (Sigma), and donkey anti-goat IgG (Santa Cruz), were used at 1: 10,000 dilutions. The optical density of the immunoreactive band was measured with a Typhoon 9000D cabinet equipped with ImageQuant software . Relative amounts of the targeted proteins were expressed as optical density (OD) relative to that of the control group of uninfected mice. If the p-tau was undetectable by Western blotting, any p-tau proteins in the brains were subjected to further quantitative analysis using an enzyme-linked immunosorbent assay (ELISA) kit (Sigma), which has the ability to detect a minimal dose of p-tau of > 7.4 pg/ml.Western blotting was performed as described by Ueberham et al.  with mod® DNA polymerase. The primers are indicated in Table Total RNA was isolated from murine brains using the GenElute™ Mammalian Total RNA Miniprep Kit (Sigma) according to manufacturer's instructions. RT-PCR was performed using a JumpStart™ RED HT RT-PCR Kit (Sigma). One microgram of total RNA was reverse-transcribed using oligo (dT)23 as the primer and enhanced avian myeloblastosis virus reverse transcriptase (eAMV-RT) (Sigma) in a 20-μl reaction mixture. The resulting complementary (c)DNA was amplified using JumpStart REDTaqP < 0.05, P < 0.01, or P < 0.001, respectively), according to a method suggested by Richardson et al. [All data of duplicate tests performed in ELISA, Western blotting, and RT-PCR were reported as the mean OD ± standard deviation (SD), and the statistical difference between the experimental group of infected mice and control group of uninfected mice could be calculated by the mean OD values obtained from ELISA, Western blotting, or RT-PCR from each experimental group of 3 mice. Those that were greater than or equal to the mean OD value plus 2, 3, or 4 SDs of the negative control group of 8 uninfected mice were considered to be statistically significant  number of wpi Fig. . All of No inflammatory cell infiltration was found in the experimental groups of infected mice at 3 dpi or 1 wpi (data not shown). Although larvae were found in or close to the choroid plexus, no leukocyte infiltration was seen in infected mice at 4 Fig.  or 8 wpiT. canis larvae invaded the mouse brains, there were no changes in the level of either TGF-β1 or S100B cytokines at 1 wpi  and 85.11 ± 0.96 ng/ml at 8 wpi (P < 0.001), both significantly higher than those found in uninfected mice (55.25 ± 2.34 ng/ml)  Fig. . HoweverRT-PCR revealed that the mRNA expression of the cytokines, TGF-β1 and S100B Fig. , glial pAβPP770, AβPP751, and AβPP695 levels significantly increased from 3 dpi to 4 wpi, followed by a significant decline at 8 wpi Fig. .T. canis is still limited. In addition, brain involvement is likely too cryptic or not easily detected in humans with CT; thus, the underlying mechanism behind the pathogenesis and sequelae of subtle BI in CT has remained largely unclear. Because it is not easily detected, the actual number of cases of subtle BI caused by this tiny parasite has probably been underestimated. Worldwide, many asymptomatic cases of toxocariasis have been confirmed to be positive by serology [Toxocara larvae embedded in internal organs may be reactivated at any time and migrate again to the brain [Although many cases of children and adults with CT clinically characterized by severe neurological disorders such as eosinophilic meningitis, encephalitis, myelitis, arachnoiditis, cerebral vasculitis, and dementia have not uncommonly been reported recently in developed countries, e.g., Austria , Belgiumserology , althoughe brain ,38. Desphe brain .T. canis infection and it seems that larvae remain in brains of BALB/c mice for a longer time compared to other strains of mice, using ICR mice as an animal model is still adequate to study CT. In fact, our previous study indicated that the larval recovery rate at a range of 2.9%~3.8% from the brain of ICR mice [T. canis eggs, the low burden of T. canis larvae  appearing in the brain and causing changes in murine behavior is likely to more realistically reflect the situation in humans and wild rodents with toxocariasis. Our present results of a mean larval burden per brain of 2.9 ± 3.0 larvae in ICR mice are close to that reported by Cox and Holland [Although Hamilton et al.  recentlyICR mice  was veryICR mice , who expICR mice  also useICR mice  also, ho Holland . In addi Holland  examined Holland  indicateT. canis larvae increasingly migrated to the brains over time, although our H&E staining showed no leukocyte infiltration or pathological changes in areas near the choroid plexus, the site of the T. canis invasion. The reason that inflammatory cell infiltration was not seen in the injured brain caused by T. canis larvae might be that the T. canis larvae mimic host tissue antigenic components and escape immune recognition or perhaps some mechanisms in the brain diminish inflammation in order to protect it from severe injuries caused by inflammatory storms [We found that y storms . Howevery storms ,44. Moduy storms ,46.Although TGF-β1 is often considered an anti-inflammatory molecule, we still propose that enhanced TGF-β1 expression may play a certain role in promoting inflammation in acute brain injury associated with experimental CT. Substantial evidence indicates that injection of an antiserum directed against TGF-β1 reduces inflammation in the CNS after traumatic injury ,48, and Elevated expression of AβPP should have deleterious effects on the acute brain injury of experimental CT. This postulation was confirmed by AβPP being involved in cerebral falciparum malaria (CFM) which causes disruption of axonal transport leading to neurological dysfunction as supported by the frequency and extent of AβPP staining being more severe in patients with than in those without CFM . It is nEnhanced expression of AβPP appears to make possible AβPP intraneuronal processing through which Aβ peptides, which are potentially detrimental to the brain, are produced . AlthougThe role of tTG induction in the neural response to injury in experimental CT is not clear. Some studies found upregulation of tTG after TBI . tTG expP. falciparum infection due to axonal injury [Similarly, it was suggested that elevated S100B levels are involved in the development of Down's syndrome and AD ; its expl injury . It was l injury . Excess l injury .Interestingly, while mRNA expressions of TGF-β1, S100B, GFAP, N F-L, tTG, and AβPP markedly declined in brains of the experimental CT mice at 8 wpi, their corresponding proteins remained abundantly present at that time. One possibility for this might be that transcription of those BIABs, which began from 4 wpi onwards, had ceased by 8 wpi. The fact that these proteins were still detectable at 8 wpi may be explained by the time lag in translating mRNAs or by the stability and persistence of the proteins. We explored the possibility that some protein degradation systems, e.g., the UPS, were disabled or impaired in brains of the experimental CT mice between 4 and 8 wpi. Ubiquitin conjugates as well as elevated ubiquitin levels in our study were found to be increased in brains of experimental CT mice, indicating impairment of the UPS's processing of excess unwanted proteins . SeveralToxocara canis larvae have been shown to be non-randomly distributed within the brain, with the telencephalon and cerebellum being their preferred sites of accumulation after at least 1 wpi, as evidenced by studies using different strains of mice inoculated with a single dose of T. canis eggs [T. canis should have potent impacts on different aspects of murine behavior. Several studies have reported varying degrees of behavior changes, and others have reported some level of memory impairment in either inbred or outbred murine hosts infected with T. canis [T. canis infection in the present study may provide more insights to better understand the pathogenesis of CT and its links with the possible development of neurodegeneration.nis eggs -68. ConsT. canis ,41,19. AT. canis . Since sT. canis -72, and T. canis , we propT. canis larval invasion of the brain. Although further studies regarding the relationship between the expressions of various BIABs and behavior disorders in experimental CT are required, results of the present study and those of the others cited in this paper suggest the possibility that cerebral infection by T. canis can have deleterious consequences and may increase the risk that CT will develop into neurodegenerative-like disease such as AD. This cannot be completely excluded because neurodegeneration is associated with the emergence of AβPP and phosphorylated tau in the brains of experimental CT mice.To the best of our knowledge, this is the first study to provide clear evidence of the concomitant presence of enhanced BIAB expression and UPS impairment during TGF-β1: transforming growth factor β1; GFAP: glial fibrillary acidic protein; NF-L: neurofilament light chain; tTGs: tissue transglutaminases; AβPPs: β-amyloid precursor proteins; UPS: ubiquitin-proteasome system; BIAB: brain injury associated biomarkers; AD: Alzheimer's disease; TBI: traumatic BI; CT: cerebral toxocariasis.The authors declare that they have no competing interests.C–KF conceived of the study, participated in the design of the study, coordination, drafted the manuscript, and performed the statistical analysis. C–KF, C–WL and W–LC carried out the molecular studies, pathological studies, and drafted the manuscript. T–CK, K–ES, and D–DJ carried out the larva recovery study, immunoassays, and gave writing suggestions. Y–HL carried out the pathological studies. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:


P=0.003) and DSS (P=0.034), both being markedly longer in tumours with negative or weakly positive staining. In multivariate Cox model, number of metastatic lymph nodes and CA IX intensity were the only independent predictors of DFS. Carbonic anhydrase isozyme IX intensity was the only independent predictor of DSS, with HR=9.2 for dying of disease with moderate-intense CA IX expression as compared with CA IX-negative/weak cases. Negative/weak CA IX staining intensity is an independent predictor of longer DFS and DSS in rectal cancer.The aim of the study is to assess the value of carbonic anhydrase isozyme IX (CA IX) expression as a predictor of disease-free survival (DFS) and disease-specific survival (DSS) in rectal cancer treated by preoperative radio- or chemoradiotherapy or surgery only. Archival tumour samples from 166 patients were analysed for CA IX expression by three different evaluations: positive/negative, proportion of positivity and staining intensity. The results of immunohistochemical analysis were confirmed by demonstrating CA IX protein in western blotting analysis. Forty-four percent of the operative samples were CA IX positive, of these 34% had weak and 66% moderate/strong staining intensity. In univariate survival analysis, intensity of CA IX expression was a predictor of DFS ( Colorectal cancer (CRC) is a common malignancy in Western countries and the incidence is rising: there were nearly 372 000 new cases of CRC in Europe in 2002  but as yα), which leads to upregulation of several genes involved in cell proliferation and angiogenesis (CA9 (CA9 encodes the carbonic anhydrase isozyme IX (CA IX)  with or without chemotherapy. Operative samples obtained from non-irradiated patients were used as controls. Carbonic anhydrase isozyme IX expression was studied in relation to histopathological features and clinical data pertinent to disease-free survival (DFS) and disease-specific survival (DSS).n=54), we studied a series of patients who had not received any treatment before surgery. After completion of the treatment protocols, all patients were followed up at the Department of Surgery.This study consists of archival operative tumour samples of 166 consecutive patients with rectal cancer, treated according to the standard protocols at Turku University Hospital. Patients in the preoperative treatment group had been operated during 2003–2008 and those in the control group between 2000 and 2008. To have a biologically and therapeutically homogenous study population, only tumours of the middle and lower rectum were included. Superficial tumours that had been treated by excision only were excluded. Standard staging included magnetic resonance imaging or computerised tomography (CT) of the rectum, CT of the abdomen and X-ray or CT of the thoracic area. Since 2005, the treatments have been planned by a multidisciplinary team. Thirty-seven patients were treated with long-course preoperative RT, generally by giving 50.4 Gy in 6 weeks, followed by surgery in about 4–7 weeks. Five of these patients were treated with 5-fluorouracil and 24 with capecitabine concomitantly with RT. Seventy-five patients were treated with short-course RT, consisting of five fractions of 5 Gy within 1 week and surgery on the following week. Post-operative adjuvant chemotherapy was considered for patients with lymph node-positive or high-risk lymph node-negative tumours. As a control group  and the National Authority for Medico-legal Affairs .Tumour regression grade was analysed after long-course RT in the HE-stained sections according to the modified Dworak and Rödel scales, using three categories: poor, moderate and excellent response  staining with rabbit polyclonal antibody for CA IX . The slides were pre-treated in microwave oven twice for 7 min in 10 mM sodium citrate buffer, pH 6. The antibody was diluted to a 1 : 8000 concentration in DAKO Antibody diluent-solution. The staining was carried out using the PowerVision+ Poly-HRP IHC-kit . The HE staining was performed according to the standard laboratory protocol.Carbonic anhydrase isozyme IX expression was analysed in all preoperative diagnostic biopsies (control group excluded) available for the study, as well as in all tumour samples obtained at operation. In each case, the most representative blocks were selected, cut to 5 The IHC stainings of the samples were evaluated by two observers (EK and JS), blinded to clinical and radiological information. Light microscopes with × 4 and × 10 objectives (EK) and × 5 and × 10 objectives (JS) were used for evaluation. Carbonic anhydrase isozyme IX staining was graded using three approaches: (1) general grouping of the cases into positive or negative, (2) proportion of positive staining and (3) staining intensity. The slides were assessed as negative, if the proportion of positive carcinoma cells in the section was less than 10%. For positive cases, the proportion of positively stained carcinoma cells was analysed using four categories: (i) 10–25%, (ii) 26–50% and (iii) over 50%. The positive slides were further evaluated to determine the staining intensity. In grading the staining intensity, three categories were used: 0 for negative, 1 for weak and 2–3 for moderate to strong staining intensity. Renal cell carcinoma was used as a positive control for strong staining intensity and normal rectal mucosa as a negative control. In weak staining intensity, there was a faint positive staining in the cytoplasm and occasional staining in the cell membranes impossible to detect at a small magnification (objectives × 4 to × 5). The staining was assessed as moderate or strong, if the positive reaction in the cell membranes could easily be identified at a small magnification (objectives × 4 to × 5). If there were areas of a variety of staining intensities, the predominant intensity was chosen. In preoperative biopsies, only positive/negative staining and staining intensity were assessed, but not the proportion of staining, because of the small size of the samples.n=4) was further studied for CA IX protein. RNA from these tumours and corresponding normal colorectal mucosa had been previously isolated for cDNA microarray analysis, and CA9 was shown to be the most upregulated gene  or Fisher's exact test for categorical variables. Differences in the means of continuous variables were analysed using Mann–Whitney's test or Kruskal–Wallis's test for two and multiple independent samples, respectively.All statistical analyses were run using SPSS  and STATA  software packages . Frequency tables were analysed using the κ and weighted κ. To calculate the latter, the ICC (intra-class correlation coefficient) test was used, with parallel mode and two-way random model, using consistency assumption and average measures option to interpret the ICC (95% CI). Concordance of CA IX expression between preoperative biopsies and operative samples was analysed using non-parametric paired-samples test (Wilcoxon signed ranks test or McNemar test). The inter-observer reproducibility of all CA IX assessments was almost perfect, with regular κ having values κ=0.988, κ=0.931 and κ=0.892, for CA IX+/−, staining proportion and intensity, respectively. Using weighted κ (ICC), even higher values were obtained: ICC=0.994, 0.985 and 0.984, respectively. This indicates that all three classifications of CA IX staining used in this study are highly reproducible.Inter-observer reproducibility of the CA IX assessments was tested using regular (Cohen's) P-value <0.05 level.Univariate survival analysis for DFS and DSS was based on the Kaplan–Meier method, where stratum-specific outcomes were compared using log-rank (Mantel-Cox) statistics. To adjust for covariates, Cox proportional hazards regression model was used, covariates being entered in stepwise backward manner. All statistical tests were two-sided and declared significant at R=0.917, P=0.0001). A low proportion of CA IX-positive staining (10–25%) was associated with weak staining intensity and vice versa. Negative, weak, moderate and strong staining intensity of CA IX has been illustrated in Carbonic anhydrase isozyme IX staining was positive in 49% of the diagnostic biopsies and 44% of the operative specimens. Staining intensity was weak in 15% and moderate or strong in 29% of the operative samples. The proportion of CA IX positive staining and staining intensity were directly related  stained for CA IX were compared with the respective operative samples (S). Using CA IX positive/negative (+/−) grading, 48 out of 78 (61%) B–S pairs were concordant and 30 out of 78 (39%) were discordant. Of the discordant samples, positive CA IX expression in biopsies was downregulated in 18 out of 38 (48%) cases, and negative CA IX expression was upregulated in 12 out of 40 (30%) cases. Using CA IX intensity evaluation 43 out of 78 (55%) B–S pairs were concordant and 35 out of 78 (45%) were discordant. In pairwise comparison (Wilcoxon), CA IX expression pattern in B–S pairs was not significantly different, P=0.006) between the groups, showing more intense staining in the group that had not received concomitant chemotherapy. Tumour regression grade was possible to be evaluated in 36 long-course RT group patients. The staining was more intense in the poor response group than in the moderate/excellent response group (P=0.010). On the other hand, CA IX expression pattern was not significantly associated with the size, nodal status or grade of the tumour, number of examined lymph nodes, resection type or its radicality, circumferential-, proximal- or distal margins, vessel invasion, tumour necrosis or patient gender.In P=0.001), as depicted in P=0.009) longer among patients who had tumours with negative or weak CA IX staining intensity (P=0.013) for disease recurrence if ⩾4 metastatic nodes and (2) CA IX intensity; HR=7.54 (95% CI 2.44–23.28) (P=0.003) for recurrence, if moderate-intense CA IX expression (CA IX-negative/weakly positive cases as reference) was present. In a similar Cox model, only CA IX intensity remained as significant independent predictor of DSS, with HR=9.23 (95% CI 2.26–37.64) for dying of disease with moderate-intense CA IX expression, as compared with CA IX-negative/weakly positive cases as reference. From the survival curves, the cumulative proportion of survivors at the 36-month follow-up time point was 83 and 35% for CA IX negative/weak and moderate/strong groups, respectively.In univariate survival (Kaplan–Meier) analysis, patients with negative or weak CA IX staining intensity had significantly longer DFS  survival model, we found that the intensity of CA IX expression was significantly related to both DFS and DSS. Patients whose tumours were negative or weakly positive in CA IX staining had better prognosis. The difference in DFS between patients with negative/weak staining intensity and those with moderate/strong staining intensity was evident from the early postoperative period, and the difference increased over time to about 50% at 3 years. This is important while considering potential interventions with adjuvant therapies. Similar data have not been previously reported in rectal cancer.When examined in multivariate (Cox) proportional hazards regression model controlled for several covariates, CA IX intensity together with the number of metastatic lymph nodes were shown to be independent predictors of DFS. Carbonic anhydrase isozyme IX intensity was the only significant independent predictor of DSS. These data implicate that intensity of CA IX expression seems to be a powerful independent prognostic factor, not confounded by the mode of treatment or other established prognostic factors (This study could not show a correlation between the expression of CA IX and other clinical or histopathological factors. Most of the patients in this study had received RT with or without chemotherapy before operation, which may have an influence on tumour size, stage and grade, nodal status and consequently, CA IX staining in the operative sample. To compensate this problem, CA IX expression was compared with the preoperative biopsies and respective surgical specimens. However, some of the available diagnostic biopsy samples were scarce, precluding adequate comparison of all sample pairs.Carbonic anhydrase isozyme IX staining intensity in the operative samples was shown to be significantly different among the treatment categories. The same was also true among patients with different response to therapy, which was analysed from the long-course RT group specimens. Most tumours in this group that had been treated with long-course RT without chemotherapy were CA IX positive with moderate/strong staining intensity. Instead, tumours that had been treated with chemoradiotherapy were mostly CA IX negative. This may imply that chemotherapy can act as a radiosensitiser and improve tumour oxygenation and consequently also final treatment outcome. This finding can presumably have important therapeutic implications in other tumour types as well.Tumour oxygenation is known to have an effect on patient prognosis. In advanced cancer of the uterine cervix, The present data are in alignment with the results reported in other human malignancies, where CA IX expression has been shown to be a prognostic marker as well . This inTo our knowledge, this is the first study to report the important prognostic significance of CA IX in rectal cancer. This study shows that strong staining intensity of CA IX is an adverse prognostic factor in rectal cancer. Further studies are needed to evaluate its potential therapeutic implications.


We investigated whether dietary intakes of total fat, monounsaturated fat (MUFA), polyunsaturated fat (PUFA) and saturated fat (SFA) were associated with breast cancer risk in a prospective cohort of 49 261 Swedish women (30–49 years at enrolment), which yielded 974 breast cancer cases by December 2005. Further, we evaluated if associations differed by oestrogen and/or progesterone receptor tumour status. Total fat, MUFA, PUFA or SFA were not associated with risk overall. However, women in the highest MUFA and PUFA quintile intake had a reduced breast cancer risk after age 50 years  compared to women in the lowest quintile. The associations did not differ by oestrogen or progesterone receptor status. Despite the negative findings, type of fat during premenopausal years may have later differential effects on risk. Animal, ecological and some case–control studies suggest that a high consumption of total dietary fat increases breast cancer risk . There i+/PR+ but negatively with ER−/PR− tumours in postmenopausal women  and saturated fat (SFA) were associated with breast cancer risk.The study cohort consisted of 49 261 women (n=244), emigration before the follow-up (n=7), very high- or low-energy intakes <1 or 99% (n=1072) and missing information on BMI, education, use of oral contraceptives (OC), age at menarche, parity, age at first birth or breast cancer in first-degree relative (mother or sister) (n=3369). Thus, the final analysis was conducted on 44 569 women. The study was approved by the ethical committee at the University of Uppsala.Around 49 261 women returned the study questionnaires and were enrolled in the study, of whom 9% were postmenopausal. We excluded 4692 women from the initial cohort due to breast cancer before enrolment  and PR status (PR− or PR+), determined by means of an Abbott immunoassay  and SFA as continuous linear variables excluding total fat. We used the standard method for energy adjustment (vs time (The models above examine the effect of substitution of fat (or fat subtypes) for carbohydrate or protein (or fat subtype) when non-alcohol energy intake is held constant. In an additional model, we also adjusted for protein and carbohydrates by fitting the following variables: non-nutritional covariates, alcohol, protein, carbohydrate, PUFA, MUFA and total fat. It examines the effect of replacing SFA with PUFA or MUFA while keeping carbohydrate, protein and total fat (and thus energy) constant. We checked the proportional hazards assumption by plotting the Schoenfeld residuals vs time .We also tested if the associations with total fat, MUFA, PUFA and SFA differed by ER and PR status. We did not conduct any analyses with joint ER and PR status because there was a strong correlation between ER and PR status.www.r-project.org).We do not have information about menopausal status after the start of follow-up. Based on the average age at menopause in Sweden being 50 years , while having an affected first-degree relative showed an increased risk .Established non-nutritional risk factors were generally evident and, for example, parous had a lower risk than nulliparous women  leaving 30 132 women with 542 breast cancer events for the analyses after age 50 years.Analysed as a continuous variable, the HR for MUFA intake was 0.55 (95% CI: 0.28–1.09) for a 10 g/day increase, while for SFA it was 1.45 (95% CI: 0.99–2.12), and for PUFA 0.58 (95% CI: 0.32–1.05) for breast cancer after age 50 years . A lineaP=0.01). When the highest PUFA quintile was compared to the lowest, the HR was 0.54 (95% CI 0.35–0.85). When we finally combined MUFA and PUFA into unsaturated fat, the HR for breast cancer was 0.79 (95% CI: 0.62–1.01) for unsaturated fat for a 10 g/day increase and 1.21 (95% CI: 1.00–1.45) for saturated fat for a 10 g/day increase. When we adjusted for protein and carbohydrate in our additional model, lower but nonsignificant breast cancer risks were found when replacing SFA with PUFA  or MUFA  for a 10 g/day increase.Compared to women in the first quintile of MUFA intake, women in the fifth quintile had a statistically significant decreased breast cancer risk  for breast cancer after 50 years of age. There was a statistically significant trend for a decreased risk across the quintiles of MUFA intake (vs ⩾50 years) and intake of MUFA, PUFA, SFA as well as unsaturated fat (PUFA+MUFA). A statistical significant interaction with age at diagnosis (<50 vs ⩾50 years) was found for intakes of PUFA (P=0.049), SFA (P=0.048) and MUFA+PUFA (P=0.040), but not for MUFA (P=0.116). These results suggest that the effects of consumption of different types of dietary fat are mainly observed in breast cancer diagnosed after age 50 years.The point estimates in For the 974 breast cancer cases, 559 (57%) were ER+, 160 (17%) were ER− and 255 (26%) had unknown ER tumour status. With respect to progesterone receptors, 474 (49%) were PR+, 235 (24%) PR− and 265 (27%) had unknown PR tumour status. A total of 94 cases had unknown ER and PR as they were registered during 2005 when no ER/PR status details were available to us. The distribution of ER+, ER−, PR+ and PR− was similar across MUFA, PUFA, SFA and total fat quintiles.vs ⩾50) for ER+ tumours (P=0.72) or PR+ tumours (P=0.85). No other statistically significant associations between total fat, MUFA, PUFA and SFA intake and risk for ER+, PR+, ER− or PR− tumours were found in the entire cohort or before or after age 50 years, irrespective of the models used (data not shown).Overall, compared to women in the first quintile of PUFA intake, women in the fifth quintile were at a decreased risk of developing ER+ tumours  and PR+ tumours  when considering the entire cohort. Similar associations were found in the fifth quintile of PUFA intake compared to the first quintile for risk after age 50 years  for ER+ tumours and: 0.52, 95% CI: 0.28–0.98) for PR+ tumours. However, there was no significant interaction between PUFA intake and age at diagnosis .The most important limitation of our study was that dietary intake was assessed only once, involving misclassification among those who changed their dietary pattern during follow-up. Because this is likely to be nondifferential, it would attenuate the strength of any true association. However, there is a potential risk that the degree of this misclassification increases over time because women change their dietary habits. Thus, the longer the follow-up, the more attenuated the association might be and we found some evidence for this: thus among women over age 50 years, the risk associated with MUFA (as a linear continuous variable) was 0.35 (95% CI: 0.13–0.97) for follow-up 0–10 years, but 1.00 (95% CI: 0.40–2.54) for longer follow-up; there was no significant interaction between MUFA and follow-up time (n-3 PUFA if these, as reported, reduce the incidence of ER− breast tumours (Women with a high PUFA intake had a decreased risk of developing ER+ and PR+ tumours after age 50 years, while no other association with fat or its subtypes differed by ER or PR status. An increased risk for ER+/PR+ tumours with higher total fat (High fat intake may increase circulating oestrogens levels . Since PStrengths of our study include its prospective design, large size and complete follow-up. Cancer registration in Sweden is obligatory, making the assessments of cases virtually complete. Furthermore, we were able to adjust for several risk factors for breast cancer. Misclassification of fat intakes due to measurement error in the food frequency questionnaire is unavoidable, but given the study design likely to be nondifferential, and thus attenuating any true association. A recent study found a null association with total fat intake using food frequency questionnaire data but a positive association when a 7-day food diary was used (Our study provides no evidence that total fat, MUFA, PUFA or SFA is associated with overall breast cancer risk, but does not rule out the possibility that types of fat may have differential effects on breast cancer risk before and after age 50 years.


The relations were examined between colorectal cancer and cigarette smoking and alcohol consumption within the Singapore Chinese Health Study, a population-based, prospective cohort of 63 257 middle-aged and older Chinese men and women enrolled between 1993 and 1998, from whom baseline data on cigarette smoking and alcohol consumption were collected through in-person interviews. By 31 December 2004, 845 cohort participants had developed colorectal cancer . Compared with nondrinkers, subjects who drank seven or more alcoholic drinks per week had a statistically significant, 72% increase in risk of colorectal cancer hazard ratio (HR)=1.72; 95% confidence interval (CI)=1.33–2.22). Cigarette smoking was associated with an increased risk of rectal cancer only. Compared with nonsmokers, HRs (95% CIs) for rectal cancer were 1.43 (1.10–1.87) for light smokers and 2.64 (1.77–3.96) for heavy smokers. Our data indicate that cigarette smoking and alcohol use interact in the Chinese population in an additive manner in affecting risk of rectal cancer, thus suggesting that these two exposures may share a common etiologic pathway in rectal carcinogenesis. Cigarette smoking and alcohol consumption are potentially modifiable lifestyle factors that have been linked to colorectal cancer risk in prospective studies of Western populations. A meta-analysis of five cohort studies conducted in the 1980s showed a statistically significant, 30% increased risk of colorectal cancer in subjects who consumed two alcoholic drinks per day .Singapore is historically a low-risk country for colorectal cancer. However, its incidence (per 100 000) has doubled among Singapore Chinese from 22.2 in 1968–72 to 42.2 in 1993–97 in males and from 16.6 in 1968–72 to 32.1 in 1993–97 in females, which are higher than comparable rates in US whites , current physical activity, menstrual/reproductive history (women only), occupational exposure, medical history and family history of cancer. Information on current diet, including alcohol consumption, was assessed via a 165-item food frequency questionnaire that has been validated against a series of 24-h dietary recall interviews  conducteFor each of the four types of alcoholic beverages , participants were asked to choose from eight frequency categories: never or hardly, once a month, 2–3 times a month, once a week, 2–3 times a week, 4–6 times a week, once a day, and two or more times a day. Consumers were then asked to choose from four defined portion sizes. For beer, the portion sizes were one small bottle (375 ml) or less, two small bottles or one large bottle (750 ml), two large bottles, and three large bottles or more. For wine, the portion sizes were one glass (118 ml) or less, two, three and four glasses or more. For Chinese or western hard liquor, the portion sizes were one shot (30 ml) or less, two, three and four shots or more. One drink was defined as 375 ml of beer (13.6 g of ethanol), 118 ml of wine (11.7 g of ethanol), and 30 ml of western or Chinese hard liquor (10.9 g of ethanol).In response to the following question, ‘’Have you ever smoked at least one cigarette a day for 1 year or longer', subjects who answered ‘no’ were classified as ‘nonsmokers’, those who answered ‘yes, but I quit smoking’ were classified as ‘former smokers’, and those who answered ‘yes, and I currently smoke’ were classified as ‘current smokers’. Ever smokers (former and current) were then asked about age at smoking initiation ; number of cigarettes smoked per day ; and duration of smoking .For physical activity, subjects were asked to estimate the number of hours spent watching TV per day, and the numbers of hours per week spent on moderate activities such as brisk walking, bowling, bicycling on level ground, tai chi or chi kung, and on strenuous sports such as jogging, bicycling on hills, tennis, squash, swimming laps or aerobics.in situ carcinomas, the analysis included 845 incident cases of invasive colorectal carcinoma, of which 516 were colon and 329 were rectal/rectosigmoid cancers.Incident colorectal cancer cases and deaths among cohort members was identified by linkage of the cohort database with the population-based Singapore Cancer Registry and Singapore Registry of Births and Deaths. The cancer registry has operated since 1968 and is comprehensive in coverage , gender, dialect group , year of recruitment , level of education , body mass index , history of diabetes , family history of colorectal cancer  and weekly physical activities . Polytomous regression methods were used to test for possible differences in exposure-risk associations by subsite (colon vs rectum)  regression methods were used to examine the associations between cigarette smoking/alcohol drinking and risk of colorectal cancer . Magnitu rectum) . AnalyseP=0.07) than duration of smoking (P=0.69) on goodness of fit, we chose the former to construct the level of smoking in combination with number of cigarettes per day. The ‘heavy’ smokers were those who started to smoke before 15 years of age and smoked 13 or more cigarettes per day; all remaining ever smokers were defined as light smokers. For the effect of smoking on risk with adjustment for alcohol consumption, a three-level alcohol index  was included as a covariate in the Cox regression models.We expressed levels of alcohol intake in units of ‘drinks’ per day to facilitate comparisons with reported data on western populations. One drink in the US is defined as one 16 oz can of beer, one glass of wine (4.5 fluid oz) or one shot (1.5 fluid oz) of hard liquor, all of which contain roughly equal amounts of ethanol (12–13 g of ethanol) . For theP-values quoted are two-sided, and P-values < 0.05 were considered statistically significant.Statistical computing was conducted using SAS version 9.1  statistical software package. All n=18 738) were ever smokers, of whom 12% (n=2304) started to smoke before age 15 and smoked at least 13 cigarettes per day (defined as ‘heavy’ smokers). The remaining 88% (n=16 434) of ever smokers started to smoke at or after 15 years of age or smoked <13 cigarettes per day (defined as ‘light’ smokers). Compared with lifelong nonsmokers, ever smokers were older, were less educated, had a lower body mass index and were more likely to report a history of physician-diagnosed diabetes  drank alcoholic beverages at least once a month, and among these, approximately 25% (n=2899) consumed seven or more drinks per week. Among them, ever smokers, on average, consumed approximately five more drinks per week than never smokers (17.3 vs 12.6 drinkers per week). Compared with nondrinkers, drinkers were better educated, had a lower body mass index and were less likely to report a history of physician-diagnosed diabetes  years and the mean time interval between entry into the study and cancer diagnosis was 5.5 years . Within the cohort, the age-adjusted incidence rate of colorectal cancer was 155 per 100 000 person-years (197 per 100 000 in men and 122 per 100 000 in women).P for trend < 0.001) after adjustment for potential confounders including cigarette smoking, physical activity and familial history of colorectal cancer. This alcohol-risk association did not differ by subsite (P=0.93). The associations between alcohol intake and disease risk were stronger in ever than never smokers, partly owing to the greater amount of alcoholic beverages consumed by the former. However, the interaction effect between alcohol and smoking was not statistically significant on risk of colorectal cancer overall and rectal cancer alone (both P>0.19), whereas the interaction effect on colon cancer risk was borderline (P=0.051).P=0.24).Beer was the main alcoholic beverage consumed, accounting for 78% of total ethanol intake, followed by western hard liquor . Consumption of wine  or Chinese hard liquor  was relatively rare. After adjustment for other sources of ethanol intake, the HR for colorectal cancer was 1.12 (95% CI=1.03–1.23) for one drink of beer per day and 1.25 (95% CI=0.98–1.59) for one drink of western hard liquor per day. The two HRs were not statistically different from each other  whereas it was stronger in nondrinkers than in drinkers .drinkers . HoweverP=0.30), colon cancer (P=0.25) and rectal cancer (P=0.92) were not statistically significant.Among 852 colorectal cancer cases, 24 (2.8%) cases were diagnosed clinically or identified through death certificates only. The results of the analyses after excluding these 24 cases were similar to those for the entire data set.Among Singapore Chinese, smoking and alcohol were independent risk factors for rectal cancer. In addition, alcohol intake also was an independent risk factor for colon cancer in this population with relatively low consumption level.The strengths of the study are its population-based design, a large study sample size, prospective data obtained via a face-to-face interview and complete cancer case ascertainment through a comprehensive nationwide cancer registry in a small city-state with a system for easy access to specialised medical care. Cigarette smoking and alcohol consumption are highly correlated lifestyle factors across ethnically and culturally distinct populations , leading to an attenuation of the estimated effects on colorectal cancer risk. Another limitation is the low alcohol consumption, especially in women, which reduces the statistical power to examine its effect on risk.vs rectum; proximal vs distal colon) (Our observed positive association between alcohol and colorectal cancer risk are consistent with the overall evidence from large, population-based Western cohorts . AlthougThe Chinese populations differ in their drinking patterns from whites, but there have been few prospective studies on the effect of alcohol on colorectal cancer, whereas a meta-analysis of 14 case–control studies in China reported an overall null association (Our finding of an increased risk of rectal cancer among smokers is consistent with cohort studies in other Asian populations (N-nitroso compounds, such as N-nitrosodimethylamine, N-nitrosodiethylamine and N-nitropyrolidine, occur in tobacco smoke (N-nitroso compounds are known inducers of colorectal carcinogenesis in experimental animals (An additive effect of alcohol and smoking on rectal cancer risk suggests that these two exposures might share a common etiologic pathway. In summary, smoking and alcohol were independent risk factors for rectal cancer in Singapore Chinese. The present study suggests that smoking and alcohol use interact in an additive manner on risk of rectal cancer and that alcohol rather than smoking is an independent risk factor for colon cancer in this population with relatively low exposure levels.


We investigated the risk of gastric cancer by subsite in relation to cigarette smoking and alcohol in a large population-based cohort of 669 570 Korean men in an insurance plan followed for an average 6.5 years, yielding 3452 new cases of gastric cancer, of which 127 were cardia and upper-third gastric cancer, 2409 were distal gastric cancer and 1007 were unclassified. A moderate association was found between smoking, cardia and upper-third (adjusted relative risk (aRR) 2.2; 95% confidence interval (CI) 1.4–3.5) and distal cancers . We also found a positive association between alcohol consumption and distal  and total  gastric cancer. Combined exposure to high levels of tobacco and alcohol increased the risk estimates further; cardia and upper-third gastric cancers were more strongly related to smoking status than distal gastric cancer. Until recently, gastric cancer was the second most common cancer worldwide; now, however, with an estimated 934 000 new cases (8.6% of new cancer cases) in 2002 alone, it is in fourth place behind lung, breast and colon and rectum cancers .The NHICS is a cohort investigation that was designed to assess the risk factors for the incidence of and mortality from cancer . We also excluded the following because of missing information: 214 for weight or height, 9936 for smoking, 3019 for alcohol intake and 6573 for dietary preference. Ultimately, 669 634 participants were included.Based on questionnaire responses at the baseline examination of the NHICS cohort, the participants were classified as ‘current smokers’ if they reported smoking currently for at least 1 year, ‘nonsmokers’ if they never smoked and ‘former smokers’ if they had smoked but quit. Current smokers were further classified by the average number of cigarettes smoked per day  and duration of smoking . Alcohol intake per day was categorised as follows: no drinking (0 g), light drinking (1–14.9 g), moderate drinking (15.0–24.9 g) and heavy drinking (⩾25.0 g). Total daily alcohol intake was expressed as the number of glasses per week of Korea's most popular alcoholic beverage, ‘Soju’. One glass of Soju contains about 12 g of ethanol. A preference for saltiness in food  was included because of possible relevance to stomach cancer .The principal outcome variable was incident gastric cancer cases identified from the KCCR, a nationwide hospital-based system that includes 94% of the country’s university hospitals and 96% of the resident training hospitals; it covers at least 90% of the newly diagnosed malignancies in Korea . Using tn=212) that were not excluded. During the 6.5-year follow-up period, we included 3452 patients diagnosed with a gastric adenocarcinoma in a final cohort comprising 669 570 participants.We used anatomic site and histological classification information from the pathology reports of the KCCR based on the International Classification of Diseases for Oncology. Tumours at the oesophagogastric junction or upper third of the stomach were classified as cardia and upper-third gastric cancer (C16.0–16.1) and those at the lower end of the stomach as distal gastric cancer (C16.2–16.7). Mixed site (C16.8) and site not otherwise specified (C16.9) were regarded as unclassified. Virtually all (>98%) of the gastric cancers were confirmed histologically, 94% being adenocarcinoma (M814–857), and the few cases . Every model included the length of follow-up as a time-dependent covariate. The proportionality assumption was verified by inspecting hazard plots. The trends were assessed by assigning ordinal values for categorical variables. Non- and former smokers were excluded from the analysis of duration and intensity (cigarettes per day) when calculating the P-value of 5% was considered significant. All statistical analyses were performed with SAS version 9.1 .The interaction effects were evaluated by calculating an interaction term, that is, multiplying a dummy variable for smoking  by one for alcohol consumption . Interactions between smoking and alcohol drinking were formally tested using the likelihood ratio method, comparing models with and without the interaction terms. We calculated a population-attributable risk  to asses−2) with 29.4% of men over 25 kg m−2. At baseline, 53.5 and 71.8% of the men were current smokers and drinkers, respectively  and had a low average BMI , cardia and upper-third cancer (P=0.68) or distal cancer (P=0.89).The independent and joint effects of smoking and alcohol intake on risks by gastric subsite are examined in Smoking and alcohol use were associated with gastric cancer risk by anatomic subsite in this large cohort study. Current smokers showed elevated risks, higher in cardia and upper-third than in distal gastric cancer among current smokers. Furthermore, we found that the risk of cancer increased with the number of cigarettes smoked per day and years of smoking. Positive associations were also found with alcohol consumption, though for cardia and upper-third gastric cancers this was not significant. The results of the multinomial logistic analysis were similar to those of a Cox proportional hazard regression: the incidence of cardia and upper-third gastric cancer was 2.2 times higher for current smokers than for never smokers, and 1.4 times for distal gastric cancer (data not shown). Combined exposure to high levels of tobacco and alcohol further increased the risk estimates.Although smoking is well recognised as a moderate risk factor, few population-based cohort studies have been conducted for gastric subsites  and their families, and consequently, may have under-represented heavy users of alcohol and tobacco. However, follow-up should be essentially complete because of our using record linkage with unique personal identifiers to national databases.Helicobacter pylori, a strong risk factor for gastric cancer (H. pylori IgG antibody among males above 40 years is 77–83% (H. pylori was observed among subjects in a longer than 5-year follow-up study in Korea (H. pylori infection (H. pylori data is unlikely to be an important issue for interpreting our findings.Third, we lacked information on c cancer . In KoreFourth, no detailed information on nutritional factors was available, including the intake of antioxidative vitamins, which might have a protective effect against gastric cancer .In our study, cardia and upper-third gastric cancer was more strongly related to smoking status than distal gastric cancer, while alcohol consumption may be associated with an increased risk of distal and total gastric cancer. Larger numbers of cardia gastric cancer, however, would be needed to investigate a dose–response relationship reliably.


Since the effect of smoking on plasma leptin has been divergent in clinical trials, which might have occurred due to selection of heterogeneous study populations, we investigated whether there is such an association in a group of healthy, non-obese, young male adults.A total of 54 smokers : 19.60±0.85) and 26 non-smokers  with similar daily physical activities and diet and without any documented disease were enrolled, and their plasma leptin levels were determined for the comparison between the two groups.2 and between 20.0 and 20.9 kg/m2 the plasma leptin levels in smokers were significantly lower when compared to non-smokers.The mean BMI and plasma leptin of smokers were significantly lower than in non-smokers. Plasma leptin in the smokers group correlated inversely with BMI and the amount of daily smoking. Below BMI 20 kg/mPlasma leptin is decreased in healthy, young non-obese male smokers independently of the amount of body fat. High amount of smoking is associated with lower serum leptin as well. Long-time smoking is characterized by reduced body-weight –3, whileSince there is a close association of body-weight with smoking and serum leptin, many authors have investigated the interaction between leptin and smoking. However, the effect of smoking on plasma leptin has been divergent in clinical trials. Some studies have mentioned decreased serum leptin in smokers –23 whileThe study was conducted on 80 young, non-obese male adults  who were allocated into two groups according to their smoking habits as 54 smokers  and 26 non-smokers . Being a smoker was defined as smoking cigarettes for at least 2 years, with more than 5 cigarettes daily. Only a ‘never-smoker’ was accepted as a non-smoker. No ex-smoker was included in any group. The participants of the study were the soldiers from a troop and had similar daily physical activities and diet for at least 6 months. None of the participants had any documented disease nor were taking any medication, including over-the-counter drugs. Physical activity levels and glucose tolerance status were similar for smokers and non-smokers.The study was approved by the Ethics Committee of Gulhane Medical School. All subjects gave written informed consent before entrance in the study.n=12) (more than 20 cigarettes a day), moderate smokers (n=12) (between 10 and 20 cigarettes a day), and mild smokers (n=30) (fewer than 10 cigarettes a day).According to the daily numbers of cigarettes, the patients were grouped as heavy smokers /height (m)Overnight fasting venous blood samples were collected at 08.00 for analysis of leptin in cold test tubes containing 0.084 mL Ethylene Diamine Tetraacetic Acid (EDTA) (0.34 mol/L). Leptin was measured by a radioimmunoassay using Human Leptin IRMA (DSL-23100 Leptin-Coated Tube Immunoradiometric Assay Kit) . Serum was stored at -30°C until analyses, and all measurements were performed in duplicate.t test was used to compare the parameters of the two groups. The associations between serum leptin levels and BMI or daily cigarette number were investigated by Spearman's correlation analysis.All data are presented as mean±SD unless otherwise noted. Student's 2 versus 21.59±1.39 kg/m2) (P <0.001). The plasma leptin levels in the smoker group were significantly lower than in non-smokers (3.47±1.1 ng/mL versus 6.63±1.5 ng/mL) (P =0.006). Plasma leptin in the smokers group correlated inversely with BMI  and the amount of daily smoking   .2  and between 20.0 and 20.9 kg/m2  of BMIs, the plasma leptin levels in smokers were significantly lower  when compared to non-smokers with the same BMI values. Since there were only two individuals in the smoker group who had a BMI above 21, comparison of plasma leptin levels in the subjects having BMIs higher than this value was not performed.The smokers and non-smokers were subanalyzed according to BMI values. Below 20 kg/mThe present study, which was designed to search for any relation between smoking and plasma leptin levels in healthy and young non-obese males, indicates that smokers have significantly lower plasma leptin levels. Low leptin levels in smokers compared to non-smokers within the similar BMI interval suggest that smoking is associated with low plasma leptin independently of the body fat content. Moreover, there was an inverse association between the number of cigarettes smoked per day and the level of plasma leptin.Leptin, which plays an important role in body-weight homeostasis through reducing food intake and increasing thermogenesis, has been studied in smokers widely. While a number of studies indicated that smoking does not affect plasma leptin levels ,24–26, sAnother possible mechanism for the lower leptin in smokers could be related to the fact that smoking increases plasma catecholamines and free fatty acid concentrations, and lipolysis which may decrease leptin concentrations –32. AlsoThe discrepant results could also be explained by other factors such as diet, exercise and other lifestyle habits, hormones, and inflammatory markers which may also regulate leptin secretion .In a recent study, leptin levels were significantly higher in women compared with men, and for both sexes were positively correlated with both BMI and the well studied marker of inflammation Creactive protein (CRP) .The link between smoking and insulin resistance syndrome was well identified ,33–35. IIn conclusion, the present study indicates that plasma leptin is decreased in healthy, young non-obese male smokers independently of the amount of body fat, and high amount of smoking is associated with lower serum leptin. Thus, smoking appears to be among the direct modulators of leptin metabolism.