Oxidatively modified LDL particles in the human placenta in early and late onset intrauterine growth restriction.

INTRODUCTION: Reduced serum LDL concentrations have been observed in pregnancies complicated by intrauterine growth restriction (IUGR) as compared to healthy pregnant women. Since increased oxidative stress has been suggested to play a major role in IUGR we now hypothesized that the lower LDL concentrations are accompanied by an accumulation of oxidized LDLs in the placenta. METHODS: Fifteen placentas of near term and preterm born IUGR, and a gestational age matched control group (CTRL n = 15) were analyzed. Placental minimal modified LDL and fully oxidized LDL particles were measured by ELISA, and by immunohistochemistry, and were related to maternal and fetal serum lipid profiles. RESULTS: We found fully oxidized LDL but not minimal modified LDL being increased in the preterm subgroup of IUGR (n = 10) as compared to preterm CTRL (n = 10; p < 0.05). An increased staining intensity of trophoblasts in preterm IUGR subjects as compared to preterm CTRL has been confirmed by immunohistochemistry (p < 0.05). No difference could be found between the term groups (n = 5 each). Correlation analysis revealed an inverse relationship of maternal LDL (ρ = −0.49, p = 0.03) and fetal HDL cholesterol (ρ = −0.46, p = 0.04) with placental fully oxidized LDL particle concentration within preterms. DISCUSSION: IUGR is a heterogeneous entity. Different pathomechanisms seem to underlie the disease in preterm and term subjects with oxidation of LDL within the placenta possibly taking place in preterm IUGRs. CONCLUSIONS: We conclude that the reduced maternal LDL cholesterol concentration in IUGR pregnancies is attributed to increased accumulation of oxidized LDL particles within the placenta at least in early onset IUGR

[Determination of maternal and foetal serum lipid profile and placental oxidised low density lipoprotein accumulation in preeclampsia and normotensive pregnancies]. / Untersuchungen des maternalen und fetalen Serumlipidprofils und des oxidierten 'Low Density' Lipoproteins im plazentaren Gewebe bei Präeklampsie und normotensiven Schwangerschaften.

BACKGROUND: Oxidised low density lipoproteins (oxLDL) are key players in the development of atherosclerotic cardiovascular diseases. Since there are similarities between the pathogenesis of preeclampsia and atherosclerosis we hypothesised an increased accumulation of oxLDL at the materno-foetal and foeto-foetal interface within the placental tissue of preeclamptic women compared to women with normotensive pregnancies (controls). Moreover, we analysed maternal and foetal serum lipid parameters. PATIENTS AND METHODS: oxLDL was determined by immunohistochemistry in placental paraffin sections of 14 women suffering from preeclampsia (30th-39th week of gestation) and compared to 28 preterm and term deliveries (25th-40th week of gestation). 10 high power fields were chosen randomly by the newCAST software and oxLDL expression was analysed via standardised methods by 2 independent and blinded investigators. Maternal and foetal triglycerides, total cholesterol, LDL cholesterol and HDL cholesterol were measured. Statistical examination was carried out by the Mann-Whitney test. RESULTS: oxLDL was found in villous trophoblast and placental endothelium. No significant differences were observed in expression intensity between preeclampsia and controls. Maternal and foetal triglyceride levels were significantly increased in preeclampsia compared to controls (pre-eclampsia mothers: 293 [SD 87.4] mg/dL, controls: 214 [SD 89.4] mg/dL, p=0.0097; preeclampsia foetuses: 26 [SD 16.6] mg/dL, controls: 18 [SD 10.4] mg/dL, p=0.0463). No significant differences in other lipid concentrations were found. CONCLUSIONS: We could not confirm our initial hypothesis of an increased oxLDL accumulation in placental tissue of preeclampsia. However, preeclampsia is a condition of dyslipidaemia affecting both maternal and foetal serum with implications for development and programming of cardiovascular diseases in later life.

PP013. Oxidized low density lipoprotein accumulation and the expression of the lectin-like oxLDL receptor (LOX-1) in placental tissue in preeclampsia and healthy controls.

INTRODUCTION: Oxidized Low Density Lipoproteins (oxLDL) and its receptor the lectin-like oxLDL receptor 1 (LOX-1) are key players in the development of atherosclerotic cardiovascular diseases. OBJECTIVES: Since preeclampsia is known to share similarities to the pathogenesis of atherosclerosis we hypothesized an increased accumulation of oxLDL and an increased expression of LOX-1 at the materno-fetal and feto-fetal interface within the placental tissue in preeclampsia in comparison to a control group. Second, we analyzed maternal and fetal serum lipid parameters including fetal oxLDL concentration. METHODS: OxLDL and LOX-1 intensity was determined via immunohistochemistry in placental paraffin sections of 11 women suffering from preeclampsia and compared to 11 gestational age matched preterm deliveries (29th to 36th week of gestation). Ten 'High Power Fields' were chosen randomly by the newCAST software and expression was analyzed via standardized methods by two independent and blinded observers. Maternal and fetal triglycerides, total cholesterol, LDL-cholesterol and HDL-cholesterol were measured by enzymatic colorimetric methods. Fetal oxLDL serum concentration was estimated by ELISA. Statistical examination was carried out by Student's t-test. Skewed variables were log-transformed. RESULTS: oxLDL and LOX-1 was predominantly found to be in villous trophoblast and placental endothelium. No significant differences could be observed in oxLDL expression intensities between preeclampsia and controls. LOX-1 expression tended to be increased in placental trophoblast and endothelium without being statistical significant (Table 1). Fetal triglyceride levels were significantly elevated in preeclampsia compared to controls while maternal triglyceride levels tend to be increased. No other significant differences in lipid concentrations could be observed (Table 2). CONCLUSION: We could not confirm our initial hypothesis of an accelerated oxLDL accumulation in placental tissue of preeclampsia. Though not statistically significant, placental endothelium seems to be activated in preeclampsia since LOX-1 expression is increased. Moreover, preeclampsia is a condition of dyslipidemia affecting both, maternal and fetal serum with implications for development of cardiovascular diseases in later life.

PP014. Estimating fully and minimal oxidized low density lipoprotein accumulation in placental tissue in intrauterine growth restriction and healthy controls.

INTRODUCTION: We recently demonstrated that maternal serum LDL- and fetal serum HDL-cholesterol concentration is significantly reduced in intrauterine growth restriction (IUGR) [1]. OBJECTIVES: We now hypothesized that increased oxidative stress in IUGR placenta leads to an accumulation of oxidized LDL (oxLDL) particles which then become trapped within the placenta subsequently leading to reduced availability of cholesterol for mother and fetus. METHODS: Fully oxidized LDL (oxLDL) was determined via immunohistochemistry in placental paraffin sections of 18 women suffering from IUGR and 18 gestational age matched controls. Ten 'High Power Fields' were chosen randomly by the newCAST software and oxLDL expression was estimated via standardized methods by two independent and blinded observers. Minimal oxidatively modified LDL (MM-LDL) and non-modified Apolipoprotein B (ApoB) concentration was measured in full placental tissue lysates by ELISA. Values were correlated with maternal and fetal total cholesterol, LDL-, and HDL-cholesterol concentrations. Statistical examinations were carried out by Student's t-test and calculation of Pearson's correlation coefficient. RESULTS: oxLDL was found predominantly to be in villous trophoblast and placental endothelium. OxLDL intensity tended to be increased in IUGR (Table 1). We found MM-LDL concentrations in whole placental tissue lysates to be highly correlated to placental ApoB concentration (r=0.93). Both parameters were non-significantly decreased in placenta of IUGR compared to controls (Table 1). Maternal serum LDL-C, and fetal serum LDL-C, TC, and HDL-C concentrations were significantly decreased in IUGR compared to controls (Table 2). OxLDL staining intensity was mildly negatively correlated to maternal LDL-C (r=-0.315) and much less to fetal HDL-C concentrations (r=-0.212). Placental ApoB and MM-LDL concentration were moderately positively correlated with fetal HDL-C concentrations (r=0.492 and r=0.447). CONCLUSION: Conformational changes of the ApoB lipoprotein during the process of oxidation might lead to an accumulation of oxLDL particles in placental tissue of IUGR and reduced fetal cholesterol bioavailability as evidenced by a decrease in fetal serum cholesterol levels. However, our analysis lacks in sufficient power and further studies are underway focussing on that subject.

Postoperative pelvic radiotherapy with or without elective irradiation of para-aortic nodes and liver in rectal cancer patients. A controlled clinical trial of the EORTC Radiotherapy Group.

OBJECTIVE: The purpose of this randomized multicenter study was to assess the impact on disease free and overall survival of low dose irradiation to para-aortic nodes and liver in patients with a locally advanced resected rectal cancer receiving a 50 Gy postoperative pelvic radiotherapy. PATIENTS AND METHODS: Main inclusion criteria were: a curative resection for a histologically proved carcinoma of the rectum, Gunderson-Sosin stages B2-B3, C1-C3, age <70 years. The patients were randomized between pelvic irradiation (Lim-XRT): 50 Gy in 25 fractions over 5 weeks and extended irradiation (Ext-XRT): same scheme/doses in the pelvis and extended fields on para-aortic nodes and liver, delivering 25 Gy in 19 fractions over 25 days. From 1983 to 1992, 484 patients were enrolled by 18 EORTC institutions and 29 patients were ineligible. The end-points were local and distant relapses, toxicity and survival. RESULTS: Compliance to treatment: 87.2% in Lim-XRT arm and 71.8% in Ext-XRT arm. Moderate acute hematological and hepatic toxicities were significantly increased in Ext-XRT arm. Among 325 patients at risk, 44 suffered a severe intestinal complication requiring surgery in 29. The 5- and 10-year estimates of disease free survival were respectively 42 and 31% in Lim-XRT arm and 47 and 31% in Ext-XRT arm (ns). The corresponding figures for overall survival were respectively 45 and 40% in Lim-XRT arm and 48 and 37% in Ext-arm (ns). The 10 years estimate of intra-pelvic failures was approximately 30% in both arms. Patients in Ext-arm appeared to have a slight shorter interval free of liver metastases (P=0.047). CONCLUSION: Low dose irradiation to the para-aortic nodes and liver did not improve survival for patients with resected adenocarcinoma of the rectum.

Selective avoidance of postoperative locoregional radiotherapy in breast cancer seems to be justified.

OBJECTIVE: To try and reduce the amount of routine postoperative radiotherapy that we prescribed without causing an unacceptable rise in locoregional recurrences. DESIGN: Retrospective study. SETTING: Teaching hospital, The Netherlands. SUBJECTS: 836 women who were treated for breast cancer between January 1980 and December 1989. INTERVENTIONS: These 836 had been treated by modified radical mastectomy (n = 534), excision of the tumour and axillary dissection (n = 279), lumpectomy (n = 15), or total mastectomy (n = 8). In December 1984 we stopped giving routine postoperative irradiation to women with T1 or T2 tumours unless there was any doubt about the operative specimen. MAIN OUTCOME MEASURES: The rate of locoregional recurrence 1985-9 compared with that from 1980-December 1984. RESULTS: Only 1 patient of 836 had a clinically detectable recurrence in the internal mammary chain. There were only 2 recurrences in the 235 axillas that had not been irradiated. CONCLUSION: By a process of careful selection of patients for locoregional irradiation, the number of fields of irradiation given to patients with breast cancer can be reduced by up to 80% without causing a rise in the rate of locoregional recurrences.

Combined external beam and low dose rate intraluminal radiotherapy in oesophageal cancer.

Thirty-five patients with oesophageal cancer were treated with external beam irradiation (50-60 Gy) followed by a boost-dose of 15-20 Gy by means of low dose rate intraluminal brachytherapy. Of the 35 patients treated 17 (48%) were pretreated with laser therapy or dilation alone. Although the intraluminal application time was long (up to 36 h) the treatment was feasible with minor acute toxicity. The palliative effect of the combined treatment was excellent; a 6 weeks post-treatment 32 of the 35 patients were able to eat solid food. Late complications were seen in six patients (17%), of which only one was severe and probably treatment-related. The median survival was 11 months; the 1- and 2-year survival were 42% and 10% respectively. The survival was strongly dependent on local control. Distant metastases became evident in 23% of patients. The interval between external radiotherapy and brachytherapy seemed to be critical. The results were compared with 68 historical controls. A significantly better survival was observed at 6 months. It is concluded that low dose rate intraluminal brachytherapy is a useful and feasible technique to increase the total dose for obtaining a better local control. The shortcomings are discussed and ideas for further improvement are mentioned.

[Combination external and internal radiotherapy for esophageal carcinoma: a new prospect]. / Combinatie van uit- en inwendige radiotherapie voor het oesofaguscarcinoom: een nieuw perspectief.

Thirty-five patients with an inoperable or non-resectable oesophageal carcinoma were treated with external radiotherapy to a maximum dose of 60 Gy followed by a booster irradiation by means of intraluminal brachytherapy at a low dose rate (40-112 cGy/h). For this extra irradiation, which lasted 20-36 h, a catheter with a thickness of 3 mm was introduced through the nose. This treatment could be carried out without sedation. The total radiation dose after the combination therapy amounted to 65-75 Gy. Of the 35 patients, 17 were pretreated with dilatation or laser therapy. The palliative effect after the external and internal irradiation was very good; only one patient still had minor dysphagia 6 weeks after the treatment. Transient irradiation oesophagitis occurred in 16 patients. In three patients, a secondary benign stenosis developed. The median duration of survival was 11 months, the 2-year survival rate was 10%. A comparison of these results with a historical control group treated only with external irradiation shows that the duration of survival of these patients appears to be increased during the first 45 weeks.

The objective response of brain metastases on radiotherapy. A prospective study using computer tomography.

A prospective study was performed to establish the objective response of brain metastases after radiotherapy. Twenty-five patients with a total of 53 metastatic lesions were evaluated approximately 6 weeks after completing the treatment (30 Gy/2,5 weeks). Clinical response was seen more often than objective response, due probably to concomitant use of corticosteroids and to most patients having multiple metastases. Clinical response, although important for the patient, does not well reflect the objective response. In 48% of the cases there was agreement between the clinical and the radiodiagnostic findings. The tumor size was an important prognostic factor for prediction of complete response. The majority of patients who achieved complete remission of at least one of the metastatic lesions, did not show recurrence of the neurological symptoms.

The choice between surgical resection and radiation therapy for patients with cancer of the esophagus and cardia: a retrospective comparison between two treatments.

The choice of treatment for patients with cancer of the esophagus and cardia is controversial. Since overall survival is poor, the most important aim of treatment should be improvement of the main complaint: the inability to eat. In a retrospective analysis of 265 patients, referred to the University Hospital in Leiden, The Netherlands, comparisons were made between palliative effects of surgical resection (N = 92) and irradiation (N = 128). Several methods of comparing surgery with irradiation are possible: (1) all surgical patients vs. all irradiated patients; (2) only those surgical patients who survived the operation (N = 70) vs. all irradiated patients (N = 128); and (3) survivors after resection (N = 70) vs. only those irradiated patients treated with "curative" radiation (N = 62). Analysis of prognostic factors showed that in both surgical and irradiated patients, the only statistically significant factor was the (dis)ability to eat. Criteria to be considered to make individual recommendations for either treatment are presented.

DNA ploidy of primary breast cancer and local recurrence after breast-conserving therapy.

The value of DNA-flow cytometry and clinico-pathological prognostic factors for the prediction of local recurrences after breast-conserving therapy (BCT) were evaluated in a retrospective study. Thirty-one patients with a local recurrence were compared with 31 matched patients without a local recurrence. Morphology and DNA-indices of the local recurrences and their corresponding primary tumours were compared. Ductal carcinoma in situ was present significantly more often in the group with a primary recurring tumour, than in the matched group (P less than 0.001), and the same holds for lobular carcinoma (n = 5). Half of the tumours that recurred had macroscopically positive surgical margins compared to about one-fourth of the matched group. Fifty-six per cent of the DNA-aneuploid stemlines in cases with local recurrence were present in the corresponding primary tumour as well (confidence limits 45%-75%), an indication that the majority of local recurrences are true recurrences and not independently developed tumours. The lack of similarity of DNA stemlines between some primary DNA-aneuploid tumours and their local recurrences indicates that these tumours had developed independently. The percentage of DNA-aneuploid cases in the group with local recurrence (89%) did not differ significantly from that in the matched group (70%). However, the findings suggest a selective recurrence of DNA-diploid stemlines. This might indicate increased resistance of DNA-diploid tumour cells to radiotherapy as compared with the resistance level in DNA-aneuploid cells.

The predictive value of cell kinetic measurements in a European trial of accelerated fractionation in advanced head and neck tumors: an interim report.

The value of cell kinetic measurements in head and neck tumors in predicting which patients will benefit from accelerated fractionation radiotherapy regimens is being tested in a multicenter European trial (EORTC trial 22851). This paper reports on the first analysis of the correlation of kinetics with outcome in this trial. A proportion of patients in both the accelerated arm (72 Gy in 5 weeks, 1.6Gy per fraction, 45 fractions) and the conventional arm (70-72 Gy in 7-8 weeks, 1.8-2.0 Gy per fraction, 35-40 fractions) were given an i.v. injection of 100 mg/m2 IUdR (iododeoxyuridine) before treatment, and a tumor biopsy was taken several hours later. The potential doubling time of the tumor (Tpot) was obtained from a flow cytometric analysis of tumor cell nuclei using an anti-IUdR antibody. From a total of 260 patients entered in the trial, 53 have undergone kinetic analysis. Adequate IUdR labeling was seen in 47 patients (88.7%), from which the mean value for Tpot was found to be 4.5 +/- 2.5 days (+/- S.D.). Of the IUdR labeled patients, 30 have now been followed up for at least 1 year, 17 with conventional and 13 with accelerated radiotherapy. These patients were split into those with fast and those with slowly growing tumors, the dividing line being the median Tpot value of 4.6 days. After conventional 7-week radiotherapy, 2 of 6 patients with "fast" growing tumors obtained local control compared with 8 of 11 with "slow" growing tumors. A small difference in local control was seen been fast and slow tumors in the accelerated arm (5/9 vs. 3/4). These preliminary data support the hypothesis that patients with fast growing tumors do poorly with conventional radiotherapy and that pretreatment kinetic measurements can select patients at risk. The predictive power of the method must await the final analysis of trial results.

Locally advanced breast cancer: radiotherapy alone versus surgery and radiotherapy.

From 1976 through 1984 46 patients with locally advanced breast cancer were treated by either radiotherapy alone or by combination of surgery and postoperative radiotherapy. Eleven of the 21 patients treated by radiotherapy alone had a local or regional recurrence, while no recurrences occurred in the combined treatment group. From this study it was concluded that "locally advanced" breast cancer is a heterogeneous disease and that further study concerning the role of surgery is justifiable for a selected subgroup of patients with non-inflammatory resectable tumours (meeting the criteria of "locally advanced" breast cancer).

The effect of radiotherapy on dysphagia and survival in patients with esophageal cancer.

A group of 127 patients with esophageal cancer treated with radiotherapy at different dose levels was retrospectively analysed. It was found that 70.5% of the patients showed improvement of dysphagia and that 54% remained palliated with respect to food passage until their death. The two major prognostic variables with respect to the palliative effect on dysphagia as well as survival were the passage score and the radiation dose. Patients with severe dysphagia (PASS 0 or 1) had a median actuarial DFI and SURV of 3.7 and 6.4 months, respectively, in contrast to 16.0 and 8.7 months for patients who were able to use (semi)solid food (PASS 2 and 3). The median actuarial DFI and SURV of patients treated with a relatively low dose (less than 50 Gy in 5 weeks) were 2.5 and 4.8 months, respectively, compared to 10.1 and 8.3 months, respectively, for patients treated with a relatively high dose.

DNA ploidy and survival in breast cancer patients.

Flow cytometric DNA ploidy measurements using frozen or deparaffinized tumor specimens were performed on 565 primary breast cancers from patients treated in the period 1975-1984. Twenty-nine percent of the cases were diploid, 61% had a single aneuploid stemline, and 10% were multiploid. Aneuploid tumors more often had negative estrogen receptor values than diploid tumors, but no significant correlation was found between ploidy class and TNM stage. Patients with more than ten positive axillary lymph nodes had predominantly aneuploid tumors. Overall and distant relapse-free survival were higher for patients with diploid tumors and low-aneuploid tumors. Stratification of the patients according to degree of lymph node involvement, TNM stage, and menopausal stage showed that the prognostic effect of aneuploidy was apparent predominantly in patients with locally advanced disease. Postmenopausal node-positive patients with diploid tumors had a significantly better prognosis than those with aneuploid tumors, but this difference was not found for the comparable premenopausal group. Multivariate analysis with the Cox proportional hazards model indicated that ploidy is an additional, independent prognostic factor in postmenopausal patients.

The prognostic significance of acute phase proteins in patients with inoperable squamous cell carcinoma of the bronchus.

In 95 patients with inoperable squamous cell carcinoma of the bronchus, nine potential serum "markers" were analysed for their prognostic significance. Lactate dehydrogenase, alpha 1 HS-glycoprotein, ferritin, carcino-embryonic antigen and immunoglobulin E did not prove to be useful as prognostic indices. The erythrocyte sedimentation rate and the acute phase proteins alpha 1 acid glycoprotein, C-reactive protein and prealbumin were correlated with survival. After taking the performance status and tumour stage into account, C-reactive protein still proved to be a strong prognosticator. The clinical relevance of the acute phase proteins signifying a so-called "biochemical status" next to the "clinical status" is discussed.

Irradiation of true pelvis for bladder and prostatic carcinoma in supine, prone or Trendelenburg position.

Small bowel transit was performed in 50 patients with bladder or prostatic carcinoma. The patients were all examined in supine and prone positions; some were also studied in 25 degrees Trendelenburg position and 25 degrees inclined procubitus to investigate the effect of the various positions on the displacement of the small bowel loops out of the true pelvis. The prone position proved to be superior to the supine position in 78% of patients. A mean displacement of 0.9 cm was obtained. Greatest shifts generally were found in the Trendelenburg position and inclined procubitus, with a mean displacement of 1.9 and 2.0 cm, respectively. The patients' height, weight, maximal abdominal circumference and Quetelet's index were analyzed with regard to the shifts of bowel loops under the various conditions. Only weight and Quetelet's index were correlated with the shifts in the Trendelenburg and inclined procubitus positions. The shifts were generally larger in case of heavier patients. We conclude that pelvic irradiation should preferably be done in the Trendelenburg or inclined procubitus position, especially in case of obesity.