Salah T. Fayed

Lipid Peroxidation and Antioxidant Status in Human Cervical Carcinoma

Reactive oxygen species (ROS), represented by superoxide, hydrogen peroxide and hydroxyl radicals, have been implicated in many diseases including cancer. ROS have been known to play an important role in the initiation and promotion of multistep carcinogenesis. The cellular antioxidants play a crucial role in protection against neoplastic disease. However, very little is known about the antioxidant defense in cervical carcinoma. This is addressed in the present study. Lipid peroxides, glutathione content and the activities of antioxidant enzymes, together with vitamin C and E content, were estimated in patients who had carcinoma of the cervix, and the values were compared with those of normal women. The results showed a remarkable reduction in the content of glutathione, vitamin E and C. Activities of glutathione peroxidase and superoxide dismutase were also reduced in cervical cancer compared to normal controls (P < 0.001). This reduction was more marked in late stages (III, IV) than in early stages (I, II) (P < 0.001). Glutathione was reduced more in poorly differentiated tumors (grade III) than in well and moderately differentiated ones (grade I, II) (P < 0.05). Levels of lipid peroxides were found to be significantly higher in malignant than in normal tissue samples and their levels were correlated with advanced clinical stage (P < 0.001). Our results suggest impaired antioxidant status in carcinoma of the cervix. This impairment is related to tumor progression.

Clinical utility of circulating matrix metalloproteinase-7 (MMP-7), CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer.

Ovarian cancer remains a highly lethal disease. The aim of the present study was to evaluate the usefulness of measuring serum matrix metalloproteinase-7 (MMP-7), CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) in comparison with serum cancer antigen 125 (CA 125) for diagnosis of epithelial ovarian cancer (EOC). This study included 51 patients with EOC, 27 patients with benign ovarian lesions and 29 healthy volunteers. Serum CA 125 was determined by microparticle enzyme immunoassay, while serum MMP-7, CCL18 and CCL11 were measured using enzyme-linked immunosorbent assay. The sensitivity and specificity were 86.3% and 92.9% for CA 125, 80.4% and 87.5% for MMP-7, 84.3% and 91.1% for CCL18 and, 68.6% and 62.5% for CCL11. Combination of CA 125, MMP-7, CCL18 and CCL11 gave a promising sensitivity of 100%, but specificity was decreased to 60.7%. The combined use of serum CA 125, MMP-7, CCL18 and CCL11 effectively detected early stages EOC with high sensitivity of 94.4%. Our data indicate that serum MMP-7, CCL18 and CCL11, in combination with CA 125 could be useful in diagnosis of EOC.

Increased bcl-2 expression is associated with primary resistance to chemotherapy in human epithelial ovarian cancer

OBJECTIVE bcl-2, an anti-apoptotic factor, has a role in the pathogenesis of ovarian cancer as well as in resistance to chemotherapy. DESIGN AND METHODS 20 benign, and 26 malignant epithelial ovarian tissues were analyzed for bcl-2 protein and mutant p53 by enzyme-immunoassay (EIA). Flowcytometric analysis was also performed. Patients of malignant group were followed up to monitor overall survival and primary resistance to chemotherapy. RESULTS bcl-2 was significantly higher in malignant group than benign group (p < 0.001). A cutoff value was determined for bcl-2 (63.8 kU/g protein). At this cutoff, sensitivity is 80.7%, and specificity is 85%. Using chi square analysis, a significant correlation was found between bcl-2 and FIGO stage (p = 0.01), overall survival (p = 0.01), as well as primary resistance to chemotherapy (p = 0.03). By correlation coefficient analysis the relation between bcl-2 and synthetic phase fraction was highly significant (p = 0.002). Bcl-2, p53, and FIGO stage were significantly correlated to poor survival (p = 0.01) in univariate analysis. However, in multivariate analysis, only FIGO stage, and p53 were independent risk factors. CONCLUSION EIA could be a useful tool for investigating the prognostic value of bcl-2, and its possible prediction of platinum resistance in epithelial ovarian cancer. This might help in selecting patients for future anti-bcl-2 therapy.

Human papillomavirus infection among Egyptian females with cervical carcinoma: relationship to spontaneous apoptosis and TNF-alpha.

OBJECTIVES This study was designed to detect HPV type-16 in Cervical carcinoma (CC) tissue specimens. The results were correlated with clinicopathological parameters of the carcinoma, with spontaneous apoptosis and with immunoreactivity to TNF-alpha antibodies. METHODS Fresh frozen tissue specimens representing 30 cases of cervical carcinoma as well as 20 normal cervical tissues (NCT) were the subjects of this study. HPV-16 DNA was detected by Polymerase Chain Reaction (PCR). The occurrence of spontaneous apoptotic cell death was analyzed by the apoptosis assay. Apoptotic cells were also counted by light microscopy and the apoptotic index (AI) was calculated. Electron microscopy was used to confirm the morphology of apoptotic cells. TNF-alpha was quantified using EIA kit. RESULTS HPV-16 DNA was more frequent in CC than in NCT. No correlation was observed between HPV infection and grade, stage or pathologic type of CC. The occurrence of spontaneous apoptosis was significantly higher in CC than in NCT, where it was correlated to advanced tumor stage and tumor pathology being more in adenocarcinoma (AC) than in squamous cell carcinoma (SCC). Moreover, AI was negatively correlated to HPV-16 infection. TNF-alpha levels were significantly higher in CC vs. NCT, where they were positively correlated to advanced tumor stage. TNF-alpha levels were correlated to DNA fragmentation and AI (r = 0.47 and 0.57 respectively). A cut-off value for TNF-alpha was calculated to be 9.1 pg/mg protein (using ROC curve). At the determined cut-off point the sensitivity was 70% and the specificity was 80%. CONCLUSIONS HPV infection, high levels of TNF-alpha and spontaneous apoptosis were strongly associated with malignant phenotype of cervical tissues. Rate of spontaneous apoptosis was higher in AC compared to SCC. On the other hand, HPV negativity was correlated with AI. Moreover, TNF-alpha and apoptotic cell death were correlated to each other as well as to tumor progression. No correlation was detected between TNF-alpha and HPV-16 infection.

The value of CA 125 and CA72-4 in management of patients with epithelial ovarian cancer.

The role of the tumor markers CA125 and CA72-4 has been evaluated in the diagnosis and management of ovarian cancer. Both markers were measured in 30 patients with proven epithelial ovarian cancer, 30 patients with benign pelvic masses and 30 normal women. CA125 and CA72-4 were measured using the luminometric immunoassay and immuno-radiometric assay respectively. All patients with ovarian cancer were submitted to surgical staging and cytoreduction followed by adjuvant platinum based chemotherapy for 3–6 courses. Fixing the specificity at 95%, CA125 had a sensitivity of 76.7% at a cut-off 85u/ml while CA72-4 had a sensitivity of 70% at a cut-off 8.5 u/ml. The combination of CA72-4 with CA125 increased the sensitivity to 95% while fixing the specificity at 95%. Among seven cases with stage I and II ovarian cancer five cases had CA125 level below 85 U/ml, three patients out of them had CA72-4 above 8.5 U/ml. CA 72-4 could reflect the residual disease following cytoreduction and could improve the detection of relapse by CA125. Conclusion: CA72-4 could complement the standard tumor marker CA125 both in diagnosis and follow up of patients with epithelial ovarian cancer.

Essential Fatty Acids Profile and Lipid Peroxides in Severe Pre-Eclampsia

There is increasing evidence that an imbalance between products of arachidonic acid metabolism (such as prostaglandins, prostacyclin and thromboxane A2) and lipid peroxidation products might have a role in the pathogenesis of preeclampsia. In order to study some of the changes taking place in pre-eclampsia, the plasma concentrations of essential fatty acids and lipid peroxidation products [measured as thiobarbituric-acid (TBA)-reactive substance] were evaluated in 25 patients with severe pre-eclampsia and 20 normal pregnant control subjects. Arachidonic acid concentration (median and range) was significantly higher in the pre-eclamptic group (14·1; 11·0–19· μg/L) compared with the normal pregnant subjects (2·5; 2·0–4·4 μg/L). Plasma concentration of TBA-reactive substance was also significantly higher in the pre-eclamptic cases (14·8; 13·4–16·8 vs 11·4; 9·0–12·8). These results suggest that arachidonic acid and lipid peroxidation products may play a role in the pathogenesis of pre-eclampsia.

Vaginal misoprostol for second-trimester pregnancy termination after one previous cesarean delivery

To determine the safety and efficacy of using misoprostol vaginally for second‐trimester abortion in women with a single previous cesarean delivery.

Local BCG injection administered to patients with flat condyloma of the cervix

⁎ Corresponding author. Department of Obstetrics and Gynecology, Faculty of Medicine, Ain Shams University, Abbassia, Cairo, Egypt, 11381. Fax: +20 2 2685 9928. E-mail address: salah_fayed@hotmail.com (S.T. Fayed). HPV infection is the most commonly diagnosed sexually transmitted viral infection [1]. The cervical lesion is typically seen at colposcopy and is known as flat condyloma. Different treatment modalities for flat condyloma range from follow-up to local excision or destruction using the large loop electrosurgical excision procedure (LEEP), laser vaporization, or cryosurgery. Half of all patients treated with LEEP return to the clinic at some point for either recurrence or appearance of a new lesion [2]. In the absence of specific antiviral treatment, the ability to enhance local immunity to overcome the viral infection is worth investigating. Local injection of BCG has long been used to treat bladder cancer with an acceptable efficacy; however, it has not been adequately investigated in the field of gynecologic oncology. We conducted a prospective, non-randomized, case control study at Ain Shams University maternity hospital, Cairo, Egypt. The study protocol was approved by the local ethical committee of Ain Shams Medical School. Informed written consent was obtained from each patient after explaining the investigative nature of this modality of treatment. Fifty patients were enrolled in the study following diagnosis of flat condyloma of the cervix based on colposcopy-guided biopsy of suspicious cervical lesion and subsequent histopathologic examination. Patients were excluded from the study if they were diagnosed with high-grade squamous intraepithelial lesion, cervical cancer, or were immunocompromised or had a history of tuberculosis. All patients underwent colposcopic examination and diagrammatic recording of the shape, area, and site of the lesion; directed biopsies from acetowhite cervical lesions were obtained. Histopathologic diagnosis of flat condyloma was established by the presence of koilocytosis. A total of 30 patients received a local cervical injection of 0.5 mL of a solution containing 0.025 mg of BCG vaccine (live attenuated, colony forming unit/mL=1–6×10; Diagsera, Cairo, Egypt). The injection was administered 1–2 weeks after biopsy. Twenty age-matched patients with cervical flat condyloma received no treatment (control group) and were followedup in the same way as the study group with colposcopic examination at 1, 3, and 6 months. Biopsy of any visible lesion was obtained at the 3-month follow-up visit. If no visible abnormality was found, a biopsy was taken from the previous site of the lesion. At the 6-month colposcopic examination a biopsy was obtained only if an abnormality was found. The demographic characteristics of the participants for age, parity, marital status, smoking, sexual partners, and use of combined oral contraceptive pills were not significantly different between the study and control groups. All patients had a single partner and combined oral contraceptives were used infrequently (6.7% in the study group). The proportion of women smoking was 10% and 15% in the study and control groups, respectively. The negative colposcopic and histopathologic findings at 1, 3, and 6 months are summarized in Table 1. In the study group, the 12 patients who remained positive on colposcopy showed a reduction in the surface area of the acetowhite lesion of more than 50%, based on the initial diagrammatic representation. Six of these 12 women showed focal rather than diffuse koilocytosis. Fever and malaise were reported by 5 (16.7%) patients in the study group, who responded to acetaminophen. The close association between HPV infection and cervical cancer has made it necessary to introduce new methods to prevent and, if possible, eradicate HPV infection of the cervix. Prevention could theoretically be achieved by immunization using the recently available vaccines. However, eradication of an established infection remains a challenge. The methods currently in use depend on local destruction of the infected tissues and are usually followed by recurrence. The method introduced in the present study aims to eliminate the virus by enhancing the local immunity of the cervix. The present study achieved a 60% negative histologic result of flat condyloma at 3 months compared with 15% in the control group. Even women with persistent disease showed both colposcopic improvement regarding the extent of the lesion and histologic improvement as the lesion became more focal and less extensive. Capello [3] investigated the effects of a local BCG injection in conjunction with interferon in patients with CIN II and III, with a response in 18 out of 20 patients. Administration of BCG in nonmuscle-invading bladder cancer has shown superiority over endoscopic resection and local mitomycin C [4]. This may encourage use of BCG in patients with high-grade squamous intraepithelial lesion if close follow-up can be guaranteed. Despite the small number of patients and the relatively short follow-up period, the preliminary results of the present study are encouraging; further studies with larger numbers and longer follow-up periods are needed.

Transforming Growth Factor-β and Nitrates in Epithelial Ovarian Cancer

The role of transforming growth factor-β (TGF-β) and nitric oxide (NO) in ovarian neoplasia is still not clear. We studied the expression of TGF-β by enzyme immunoassay, and nitrates (as a stable end product of NO) in 127 ovarian tissues (36 normal, 37 benign, and 54 malignant). Ploidy status and synthetic phase fraction (SPF) were also assessed by flow cytometry. Mean ranks of TGF-β, nitrate, and SPF were significant among different groups (X2 = 12.01, P = 0.0025, X2 = 67.42, P = 0.000, X2 = 9.06, P = 0.011 respectively). Nitrate mean ranks were significant among different FIGO stages of the disease (X2 = 17.6, P = 0.000). A significant correlation was shown between TGF-â, and nitrate levels in all tissues (r = 0.24, P = 0.01), as well as in malignant tissues (r = 0.3, P = 0.026). Cutoff values were determined for both TGF-β (290 pg/mg protein), and nitrates (310 nmole/mg non protein nitrogenous substances). At these cut-offs, nitrates showed a sensitivity of 93% and 84% specificity for malignant versus normal cases, while TGF-β had 76% sensitivity, and 82.4% specificity for poor versus good outcome. Patients with epithelial ovarian cancer were followed up for a total of 40 months. Survival analysis showed that patients with TGF-β above the cut-off had worse prognosis (X2 = 12.69, P = 0.004). The present results suggest that malignant transformation of ovarian tissues is associated with increased TGF-β and NO production. NO level is related to the development and progression of epithelial ovarian cancer, while high levels of TGF-β could be of prognostic significance.

The predictive value of C-erbB-2 and p53 overexpression in the management of molar pregnancy

Objective To test the usefulness of immunostaining for the tissue markers C-erbB-2 and p53 in predicting the course of the disease in molar pregnancy in spontaneous remission or development of gestational trophoblastic neoplasia. Methods A total of 71 cases, comprising a partial mole (34 cases), a complete mole (33 cases), an invasive mole (two cases), and choriocarcinoma (two cases), were analyzed immunohistochemically on paraffin blocks using anti-p53 and anti C-erbB-2 antibodies. The clinical data were retrieved from the follow-up sheets of these patients and correlated with the results of immunostaining. Results The results showed that the expression of p53 and C-erbB-2 was markedly enhanced in the gestational trophoblastic diseases. There was a trend toward a higher expression of protein p53 in cases of complete moles, invasive moles, and choriocarcinoma, and a trend toward a negative expression in cases of partial moles. The two markers together had a poor positive predictive value but a good negative predictive value. Conclusion Positive immunostaining for p53 and C-erbB-2 could not predict the requirement for chemotherapy in patients with molar pregnancy, whereas the concomitant negative staining for both markers could predict spontaneous remission.