Profiling of Discrete Gynecological Cancers Reveals Novel Transcriptional Modules and Common Features Shared by Other Cancer Types and Embryonic Stem Cells.

Studies on individual types of gynecological cancers (GCs), utilizing novel expression technologies, have revealed specific pathogenetic patterns and gene markers for cervical (CC), endometrial (EC) and vulvar cancer (VC). Although the clinical phenotypes of the three types of gynecological cancers are discrete, the fact they originate from a common embryological origin, has led to the hypothesis that they might share common features reflecting regression to early embryogenesis. To address this question, we performed a comprehensive comparative analysis of their profiles. Our data identified both common features (pathways and networks) and novel distinct modules controlling the same deregulated biological processes in all three types. Specifically, four novel transcriptional modules were discovered regulating cell cycle and apoptosis. Integration and comparison of our data with other databases, led to the identification of common features among cancer types, embryonic stem (ES) cells and the newly discovered cell population of squamocolumnar (SC) junction of the cervix, considered to host the early cancer events. Conclusively, these data lead us to propose the presence of common features among gynecological cancers, other types of cancers, ES cells and the pre-malignant SC junction cells, where the novel E2F/NFY and MAX/CEBP modules play an important role for the pathogenesis of gynecological carcinomas.

Low mutational burden of eight genes involved in the MAPK/ERK, PI3K/AKT, and GNAQ/11 pathways in female genital tract primary melanomas.

Mucosal melanomas exhibit discrete genetic features compared to cutaneous melanoma. Limited studies on gynecological melanomas revealed significant heterogeneity and low mutational burden. To gain further insight into their genetics and DNA repair efficiency, we systematically investigated the status of eight genes whose products are critically involved in the MAPK/ERK, PI3K/AKT, and GNAQ/11 pathways, including BRAF, NRAS, HRAS, KRAS, c-KIT, PI3K, GNAQ, and GNA11, in a series of 16 primary gynecological melanomas, covering all anatomical locations, ranging from stages I to III. Analysis either by real-time PCR coupled with fluorescence melting curve analysis or by PCR followed by direct sequencing, along with studies for DNA mismatch repair status using immunohistochemistry, disclosed that 15 out of the 16 cases displayed wild-type genotypes, with a single case of vulvar primary melanoma, harboring the activating mutation BRAF(V600E). Investigations on whether this could reflect partly an efficient mismatch repair (MMR) mechanism were confirmed by normal expression of hMLH1 and hMSH2, suggesting that the lack of mutations could be explained by the operation of alternative pathogenetic mechanisms modulating downstream effectors of the signaling pathways. Our data suggest the presence of additional genetic components and provide the impetus for systematic approaches to reveal these yet unidentified genetic parameters.

Expression profiling of vulvar carcinoma: clues for deranged extracellular matrix remodeling and effects on multiple signaling pathways combined with discrete patient subsets.

The molecular mechanisms of vulvar squamous cell carcinoma (VSCC) remain obscure. To this end, we investigated systematically for the first time the expression profile of VSCC using the microarray technology, in a total of 11 snap-frozen samples, from five VSCC patients covering early and advanced stages of VSCC undergoing radical surgery and from six matched healthy controls. All experiments were performed using Affymetrix Human Genome U133A 2.0 oligonucleotide arrays, covering 22,277 probe sets. Genes were filtered and analyzed using analysis of variance, t test, fold-change calculations, and unsupervised hierarchical cluster analysis. Further processing included functional analysis and overrepresentation calculations based on Gene Ontology, Database for Annotation, Visualization, and Integrated Discovery, and Ingenuity Pathway Analysis. The molecular phenotypes of VSCC patients exhibited significant and discrete transcriptional differences from the healthy controls, whereas principal component analysis documented that this separation is mediated by a consistent set of gene expression differences. We detected 1077 genes (306 upregulated and 771 downregulated) that were differentially expressed between VSCC patients and healthy controls by at least twofold (P < .01), whereas a novel subset of patients was revealed displaying a distinct pattern of 125 upregulated genes involved in multiple cellular processes. Functional analysis of the 1077 genes documented their involvement in more than 50 signaling pathways, such as PTEN, oncostatin M, and extracellular signal-regulated kinase signaling, affecting extracellular matrix remodeling and invasion. Comparison of our data set with those of the single VIN study revealed that the two entities share a limited number of genes and display unique features.

Maternal-neonatal magnesium and zinc serum concentrations after vaginal delivery.

BACKGROUND/AIMS: To investigate the effect of the mode of delivery on maternal-neonatal Mg and Zn levels. MATERIAL AND METHODS: Two groups of pregnant women participated in the study: Group A (n = 16) with normal labor and vaginal delivery and group B (n = 14) with scheduled cesarean section (CS). Blood was obtained at the beginning of the labor, immediately after delivery and from the umbilical cord (CB). Serum Mg and Zn were measured with atomic absorption spectroscopy and total antioxidant status (TAS) levels with a chemical autoanalyser. RESULTS: Mg, Zn and TAS levels were similar pre-delivery in both groups. TAS levels, Mg (0.81 ± 0.09 vs 0.69 ± 0.03 mmol/L, p < 0.001) and Zn levels (9.34 ± 0.37 vs 5.74 ± 0.24 µmol/L, p < 0.001) were significantly decreased after vaginal delivery. These biochemical parameters were measured practically unaltered at the same times of study in group B. The mineral levels did not differ in the CB of both groups. CONCLUSIONS: The decreased maternal Mg, Zn and TAS levels post vaginal delivery may be due to the participation of skeletal and uterus muscles and the similar levels of the minerals in the CB of neonates to the placental protection.

p16 INK4A is a strong biomarker for cervical intraepithelial neoplasia and invasive cervical carcinoma: a reappraisal.

To assess the role of p16( INK4A), bcl-2, and p53 in cervical cancer screening, we conducted a retrospective trial of prospectively collected data. Sixty two women with abnormal Pap smears were subjected to colposcopy and biopsies from any abnormal lesion. Human papillomavirus (HPV)-DNA typing, histology, and immunochemistry for p16(INK4A), bcl-2, and p53 were performed for these women. Histologic diagnosis was that of low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL), or cancer in all cases. Human papillomavirus strains were identified in 56 patients (90.3%). All HSIL and invasive cancer cases were HPV positive. p16(INK4A) immunostaining yielded 100% sensitivity, 76% specificity, 61% positive predictive value, and 100% negative predictive value in cancer patients. The corresponding performance indicators for HSIL patients were 75%, 62%, 32%, and 91%. bcl-2 and p53 expression did not correlate with worsening grades of cervical disease. We conclude that p16(INK4A) seems to be a sensitive biomarker of high grade cervical intraepithelial neoplasia and cancer.

Maternal-neonatal amino acid blood levels in relation to the mode of delivery.

OBJECTIVE: To investigate the effect of the mode of delivery on maternal-neonatal amino acid levels as high blood levels of some amino acids are implicated with endurance exercise. DESIGN: Comparative study. SAMPLE: Thirty women in normal pregnancy divided into two groups: Group A (n=15) with normal labor and vaginal delivery and group B (n=15) with scheduled cesarean section. MATERIAL AND METHODS: Blood was obtained from the mothers pre- versus post-delivery as well as from the umbilical cord. Routine laboratory tests (liver enzymes, muscle enzyme, etc.) and the amino acid blood levels were measured with a clinical chemistry analyzer and tandem mass spectrometry methods, respectively. RESULTS: Routine laboratory tests and the amino acid blood levels were similar in the two groups of mothers pre-delivery. Total antioxidant status levels were reduced, whereas the branched-chain amino acids (BCAAs) and alanine levels were remarkably elevated in the sera of group A post-delivery, whereas they remained unaltered in group B at the same time of study. The mentioned BCAAs and alanine levels were higher in the umbilical cord blood of group A than those in group B. The rest of the amino acids were similar. CONCLUSIONS: The increased BCAAs and alanine blood levels in mothers of group A may be related to uterine and skeletal muscle contractions during the vaginal delivery process and the high levels in the umbilical cord blood of their neonates may mirror those of the mothers. The elevation of BCAAs both in mothers of group A and their neonates may exclude or minimize tyrosine and tryptophane levels from entry in the brain resulting in decreased biogenic amine and increased prolactin production in the central nervous system of these mothers and their infants.

The effect of the mode of delivery on maternal-neonatal interleukin-6, biogenic amine and their precursor amino acid concentrations.

BACKGROUND: Biogenic amine, adrenaline, noradrenaline, dopamine and 5-hydroxy-tryptamine (5-HT) levels are related to interleukin-6 (IL-6) plasma concentrations and endurance exercise. The aim of our study was to investigate the effect of the mode of delivery on maternal-neonatal IL-6, biogenic amine and their precursor amino acid levels. METHODS: Some women with normal pregnancy (n=56) were divided into two groups: group A (n=26) with normal labor and vaginal delivery, and group B (n=30) with scheduled cesarean section. Blood was obtained from the mothers at the beginning of labor and immediately after delivery (pre- vs. post-delivery), as well as from the umbilical cord (CB). Total antioxidant status (TAS) and IL-6 levels were measured with commercial kits, the precursor amino acids, tyrosine and tryptophan with tandem mass spectrometry and the biogenic amine blood levels with HPLC methods, respectively. RESULTS: TAS, IL-6, biogenic amine and their precursor amino acid blood levels were similar in the two groups of mothers pre-delivery. TAS levels were reduced, whereas the amino acids, the catecholamine, 5-HT and IL-6 levels were increased post-delivery and in the CB of group A and unaltered in group B at the same time of the study. CONCLUSIONS: During a vaginal delivery process, the low TAS, the increased levels of the studied amino acids, the catecholamines, 5-HT and IL-6 may be due to the activation of the neuroendocrine system and the participation of skeletal and uterine muscles. The mode of delivery may be taken into account when IL-6 plasma levels are evaluated as an anti-inflammatory index perinatally.

The effect of the mode of delivery on the maternal-neonatal carnitine blood levels and antioxidant status.

BACKGROUND: Carnitine blood levels are closely related to beta-oxidation and implicated with strenuous muscle contractions. Normal delivery process is characterized by the participation of the uterus and most skeletal muscles. METHODS: Women with normal pregnancy (n = 56) were divided into two groups. Group A (n = 26) with normal labor and vaginal delivery and group B (n = 30) with scheduled cesarean section. Blood was obtained from the mothers at the beginning of labor and immediately after delivery (pre- vs. post-delivery), as well as from the cord blood (CB). Total antioxidant status (TAS) was measured with a commercial kit and carnitine was measured in blood spots on Guthrie cards with tandem-mass spectrometry. RESULTS: TAS and carnitine levels were similar in all the groups pre-delivery. In contrast, TAS and carnitine levels were significantly lower in group A than in group B post-delivery. Remarkably lower TAS and carnitine levels were measured in the CB of neonates of group A as compared to the CB of neonates of group B. CONCLUSIONS: The lower TAS and carnitine levels measured in group A as compared to group B postdelivery may be due to uterus and skeletal muscle contraction during a normal labor process. Infants born with scheduled cesarean section are benefited with high carnitine levels to face oxidation perinatally.

The effect of the mode of delivery on the maternal-neonatal erythrocyte membrane acetylcholinesterase activity.

UNLABELLED: Free radical production and high catecholamine levels are implicated with the modulation of acetylcholinesterase (AChE) activity. OBJECTIVE: To investigate the effect of the mode of delivery on maternal-neonatal erythrocyte membrane AChE activity. SUBJECTS AND METHODS: Some women with normal pregnancy (N = 30) were divided into two groups: group A (N = 16) with normal labour and vaginal delivery and group B (N = 14) with scheduled Cesarean section, twenty non-pregnant women were the controls. Blood was obtained from controls and from mothers pre- vs post-delivery as well as from the umbilical cord (CB). Total antioxidant status (TAS), membrane AChE activities and catecholamine blood levels were measured with a commercial kit, spectrophotometrically and HPLC methods, respectively. RESULTS: TAS and catecholamine levels as well as membrane AChE activities were similar in the two groups of mothers pre-delivery and in controls. TAS levels were reduced whereas AChE activities and catecholamine levels were increased post-delivery in mothers of group A and unaltered in group B at the same times of study. AChE activity was similarly lower in the CB of neonates than those of their mothers pre-delivery. CONCLUSIONS: During a normal delivery process, the low TAS, the increased levels of catecholamines and the increased AChE activity, post-delivery, may be due to the increased stress due to the participation of uterus and skeletal muscles as during endurance exercise. The low AChE activity in newborns may be related to perinatal immaturity.

Maternal-neonatal erythrocyte membrane Na(+), K (+)-ATPase and Mg (2+)-ATPase activities in relation to the mode of delivery.

Free radical production and high catecholamine levels are implicated in the modulation of Na(+), K(+)-ATPase, and Mg(2+)-ATPase activities. The aim of this study was to investigate the effect of the mode of delivery on the above-mentioned enzyme activities in maternal-neonatal erythrocyte membrane. Women with normal pregnancy (N = 30) were divided into two groups: Group A (N = 16) with normal labor and vaginal delivery, and Group B (N = 14) with scheduled cesarean section; 20 non-pregnant women were the controls. Blood was obtained from controls and mothers, pre- versus post-delivery, and from the umbilical cord (CB). Total antioxidant status (TAS), membrane enzyme activities, and catecholamine blood levels were measured with a commercial kit, spectrophotometrically, and by HPLC methods, respectively. The results showed that: TAS levels, catecholamine, and the membrane enzyme activities were similar in the two groups of mothers pre-delivery, whereas both enzyme activities were lower than those of controls. TAS levels were reduced whereas Na(+), K(+)-ATPase activities (0.35 +/- 0.03 vs. 0.65 +/- 0.06 micromol Pi/h x mg protein, P < 0.001), and catecholamine levels were increased post-delivery in mothers of Group A and unaltered in Group B (0.38 +/- 0.02 vs. 0.40 +/- 0.03 micromol Pi/h x mg protein, P > 0.05), at the same times of study. Mg(2+)-ATPase activities remained unaltered in both groups of mothers and newborns. Na(+), K(+)-ATPase activity was similarly lower in the CB of neonates than those of their mothers, pre-delivery. Our results suggest that: (a) during a normal vaginal delivery process, the low TAS and the increased levels of catecholamines may increase Na(+), K(+)-ATPase activity, post-delivery; (b) the low enzyme activities evaluated in mothers pre-delivery may be due to the high estrogen levels and those in newborns due to perinatal immaturity.

Maternal chronic hepatitis B virus does not affect neonatal biotinidase activity.

BACKGROUND: Biotinidase activity is closely related to liver function. AIM: To evaluate whether maternal chronic hepatitis B virus (HBV) infection affects neonatal biotinidase activity. PATIENTS AND METHODS: Twenty-three asymptomatic pregnant women with HBV (group A) and 28 healthy pregnant women (controls) in the delivery room and their newborns (cord blood) underwent laboratory examinations. Serological HBV and liver function tests were performed with standard techniques, while biotinidase activity was measured with an HPLC method. RESULTS: Serological HBV tests and HBV DNA showed chronic HBV (precore mutant G1896A) in group A, whereas anti-HBc and anti-HBe were detected in their neonates. Liver function chemistry was found normal in controls and both groups of newborns. Moderately increased transaminases were found in the infected mothers. Interestingly, albumin levels did not differ among the studied groups. Biotinidase activity in HBV mothers (5.76+/-0.6 nmol/min/mL) was significantly decreased (p<0.001) as compared to controls (8.43+/-0.65 nmol/min/mL). The enzyme activity did not differ among the neonates. Biotinidase activity inversely correlated with transaminases but not with albumin or with HBV-DNA levels. CONCLUSIONS: Decreased biotinidase activities were evaluated in mothers with HBV and normal in their neonates. Biotin supplementation in the diseased mothers may prevent possible symptoms due to biotin recycling impairment.

Maternal chronic hepatitis B virus is implicated with low neonatal paraoxonase/arylesterase activities.

BACKGROUND: Paraoxonase/arylesterase activities are closely implicated with liver function and antiatherogenetic process. AIM: To evaluate whether maternal chronic hepatitis B virus, disease (HBV) affect serum neonatal paraoxonase/arylesterase activities. PATIENTS AND METHODS: 28 pregnant women with HBV and 28 healthy pregnant women (controls) in the delivery room and their newborns (cord blood) underwent laboratory examinations. Serological virus tests and liver function tests and paraoxonase (PON 1) activities were measured with the Siemens Advia 1650 Clinical Chemistry System, while total antioxidant capacity (TAC) levels and paraoxonase-arylesterase (PON-aryl) activities were measured spectrophotometrically. RESULTS: Serological HBV tests and HBV DNA showed chronic HBV (precore mutant G1896A) in the diseased mothers whereas anti-HBc and anti-HBe were detected in their neonates. Liver function parameters were found normal in controls and both groups of newborns. Moderately increased transaminase levels were measured in HBV mothers, whereas TAC levels were decreased in hepatic mothers and their newborns. Interestingly albumin levels did not differ among the studied groups. PON 1 and PON-aryl activities in the diseased mothers (148+/-14 U/mL/min, 130+/-16 KU/mL/min) and their infants (32+/-6 U/mL/min, 24+/-5 KU/mL/min) were significantly lower as compared to those of control mothers (217+/-16 U/mL/min, 196+/-14 KU/mL/min p<0.001) and their newborns (57+/-6 U/mL/min, 48+/-8 UK mL/min p<0.001). Inverse significant correlations were found between the studied enzyme activities and liver enzymes in all the groups of study except in infants born from HBV mothers and positive with TAC in all the studied groups. CONCLUSIONS: Decreased PON 1 and PON-aryl activities were measured in infants born from hepatic mothers probably as a consequence of their low TAC. Infants born from HBV mothers are at risk for developing LDL oxidation perinatally.

Maternal-neonatal 8-hydroxy-deoxyguanosine serum concentrations as an index of DNA oxidation in association with the mode of labour and delivery.

AIM: To investigate the effect of the mode of labour and delivery on the total antioxidant status (TAS), and the biomarker of DNA oxidation, 8-hydroxy-deoxyguanosine (8-OHdG) serum levels, in mothers and their newborns. SUBJECTS AND METHODS: Some 106 women with normal pregnancy and normal blood biochemical parameters were divided into 4 groups: Group A (n=28) with normal labour and vaginal delivery (VG), Group B (n=25) with scheduled cesarean section (CS), Group C (n=26) with 'emergency' CS, and Group D (n=27) with prolonged labour+VG. Blood was obtained from the mothers at the beginning of labour, and immediately after delivery (pre- and post-delivery), as well as from the umbilical cord (CB). TAS, 8-OHdG and creatine kinase (CK) were measured in the sera with appropriate methodology. RESULTS: TAS levels were almost similar in all the groups pre-delivery, and in CB irrespective of the mode of labour and delivery, and remarkably decreased in Groups C and D post-delivery. 8-OHdG levels in Group C (0.94+/-0.08 ng/ml) and Group D (0.98+/-0.08 ng/ml) were significantly higher than those in Group A (0.26+/-0.01 ng/ml, p<0.001) and Group B (0.28+/-0.07 ng/ml, p<0.001) post-delivery. 8-OHdG levels were low in CB, independent of the mode of labour. CK positively correlated with 8-OHdG (r=0.48, p<0.001), the latter negatively correlated with TAS (r=-0.53, p<0.01). CONCLUSIONS: The lowest TAS and the highest 8-OHdG levels were found in Groups C and D post-delivery, probably due to the long-term participation of the mothers' skeletal and uterus muscles, whereas 8-OHdG levels were low in CB irrespective of the mode of delivery, possibly as a consequence of the antioxidant action of the placenta and/or the low lipid levels in the serum of the umbilical cord.

Maternal-neonatal serum paraoxonase 1 activity in relation to the mode of delivery.

AIM: To investigate the effect of the mode of labour and delivery on the total antioxidant status (TAS) and the paraoxonase 1 (PON 1) serum activity in mothers and their newborns. SUBJECTS AND METHODS: One hundred six women with normal pregnancy were divided into 4 groups: group A (n = 28) with normal labour and vaginal delivery (VG), group B (n = 25) with scheduled caesarean section (CS), group C (n = 26) with "emergency" CS and group D (n = 27) with prolonged labour + VG. Blood was obtained from the mothers at the beginning of the labour process and immediately after delivery (pre- and post-delivery) as well as from the umbilical cord (CB). PON 1 activity and blood chemistry were determined using the Bayer Advia 1650 Clinical Chemistry System, whereas TAS levels were measured spectrophotometrically at 450 min in microtiter plates. RESULTS: TAS levels were similar pre-delivery and low in CB in all the groups. In contrast, TAS levels were remarkably reduced in group C and in group D post-delivery whereas they were nearly unchanged in group B and just lowered in group A, at the same time of study. PON 1 activity was practically unaltered in group A and group B pre- vs. post-delivery. Interestingly, the enzyme activity was remarkably decreased in group C (222 +/- 16 vs. 153 +/- 14 U/min/mL) and group D (216 +/- 16 vs. 135 +/- 15 U/min/mL, p < 0.001) as compared with those of the other groups at the same time of study. Additionally, PON 1 activity was higher in the newborns of group A and group B than those in group C and group D. TAS and HDL positively correlated with PON 1 activity. CONCLUSION: The low TAS levels and the decreased PON 1 activity, which were found in groups C and D post-delivery, may be due to the increased production of free radicals, during long-lasting labour + VG and obstructive labour + CS. PON 1 activity was low in CB irrespectively of the mode of delivery, probably due to the low lipid levels in the serum of the umbilical cord. Neonates born with normal delivery or scheduled CS are benefited with a higher antiatherogenic enzyme activity perinatally.