Vesna Čapkun

Maternal and Neonatal Effects of Substance Abuse during Pregnancy: Our Ten-year Experience

Purpose The aim of the study was to assess perinatal outcome of pregnancy burdened with maternal addiction in comparison with an unselected population from a European transition country. Materials and Methods Data on pregnancies complicated by illicit drug abuse (n = 85) managed during a 10-year period (1997 - 2007) at Split University Hospital were analyzed. Data on the type of drug, course of gestation and labor, and on perinatal outcome were considered. Data on all non-dependence pregnancies recorded during the study period were used as a control group. Results During the study period, there were 85 dependence-complicated pregnancies (0.2%). Use of heroin alone during pregnancy was recorded in 51 women (50%), methadone alone in 6 (7%), and a combination of heroin and methadone in 9 (11%). Premature delivery was significantly more common in the group of pregnant addicts (21% vs. 6%); 49% of pregnant addicts were carriers of hepatitis C virus (HCV) and 14% of hepatitis B virus (HBV). Neonatal abstinence syndrome developed in 61 infants (7%) born to addicted mothers. There were 4 cases (4.6%) of early neonatal death; 7 neonates had 5-minute Apgar score ≤ 7 (8%); 29 neonates had low birth weight for age (33%); and 7 neonates had congenital anomalies (8%). The risk of various congenital anomalies was 3-fold in the group of children born to addicted mothers. Conclusion Addiction pregnancies present a small but high-risk group according to perinatal outcome. Appropriate obstetric and neonatal care can reduce the rate of complications in these pregnancies and improve perinatal outcome.

Expression of androgen receptors in triple negative breast carcinomas.

Triple negative breast cancer (TNBC) consists of a group of tumors with poor prognosis, owing to aggressive tumor biology and lack of targeted therapy. The aim of this study was to assess the immunostaining for androgen receptors (ARs) in the group of TNBC, in addition to basal-like (BL) immunophenotype, BL morphology and conventional clinicopathological factors and to demonstrate its prognostic relevance in this group of tumors. The study included 83 patients. Slides were stained immunohistochemically for estrogen and progesterone receptors, HER2, CK5/6, CK14, EGFR, Ki-67 and AR. Of the 83 TNBC samples, 32.5% showed positive immunostaining for AR, 66.3% had BL immunophenotype, and 48.2% had BL morphology. Positive AR immunostaining was inversely correlated with higher clinical stage, higher mitotic score, higher histological grade and higher proliferation index measured by Ki-67. Significantly more AR negative tumors were observed among the tumors with BL immunophenotype and BL morphology. There was no significant association between positive AR immunostaining and disease free survival or overall survival. More than one third of TNBC were AR-positive, and this represents a potential opportunity for novel targeted treatment in the group of breast tumors for which therapeutic options are currently limited.

IMP3 can predict aggressive behaviour of lung adenocarcinoma

BackgroundLung cancer most often presents as an inoperable tumour and the diagnosis is usually performed on a small biopsy/cytology specimen. In the group of non small cell lung cancer - not otherwise specified, adenocarcinoma phenotype can be determined immunohistochemically using TTF-1 and Napsin A. Expression of oncofetal protein IMP3 in human cancer is associated with poor differentiation and aggressive behaviour. In the present study expression of IMP3 was correlated with expression of TTF-1 and Napsin A, histological subtype and clinical stage of lung adenocarcinoma. We were interested whether distant metastases are associated with IMP3 overexpression, regardless of the histologic subtype of adenocarcinoma.MethodsIn retrospective study, consecutive series of 105 patients with advanced lung adenocarcinoma diagnosed from 2006 to 2009 in Clinical Hospital Center Split, Croatia, were analysed. Clinical data were collected from the Pulmology Department and time of death from the Mortality Registry. Paraffin blocks of bronchoscopic biopsies were collected from the Institute of Pathology and 15 cases excluded from the analysis due to insufficient material. Expression of IMP3, Napsin A and TTF-1 were analysed by indirect enzyme immunohistochemistry. Statistical analysis was performed and P values less than 0.05 considered significant.ResultsOf 90 patients, 71 (78%) were males and 19 (22%) females. Median age for males was 61.5 years (min-max 43–83) and for females 61 years (min-max 44–86). Pleural effusion was found in 15 (16.6%) and distant metastases in 45 (50%) cases. According to histological subtypes, there were 34 acinar, 2 lepidic, 2 papillary and 52 solid subtypes. IMP3 overexpression was found in 63 cases (70%) and was correlated with solid subtype (P = 0.002) and negative/weak Napsin A expression (P = 0.004). Strong Napsin A expression correlated with TTF-1 expression (P = 0.003) and lower histological grades (P = 0.031). Patients with IMP3 overexpression more often had distant metastases than patients with negative IMP3, 55.5% versus 33.3% (P = 0.033). Non solid subtypes with IMP3 overexpression developed distant metastasis more common than non solid subtypes with negative IMP3, 72% versus 35% (P = 0.028).ConclusionsExpression of IMP3 correlates with solid subtype and with distant metastases regardless of histological subtype of lung adenocarcinoma.Virtual slideshttp://www.diagnosticpathology.diagnomx.eu/vs/1966211581795258ZusammenfassungHintergrundDas Lungenkarzinom kommt meistens als nicht resektabler Tumor vor und die Diagnose kann nur in kleinen Biopsaten oder zytologisch gestellt werden. In der Gruppe der nicht kleinzelligen Lungenkarzinome kann der nicht anders spezifizierte Adenokarzinom Phänotyp mit Hilfe der Antikörper TTF-1 und Napsin A diagnostiziert werden. Die Expression des onkoföetalen Proteins IMP3 ist bei humanen Karzinomen mit agressivem Verhalten und metastatischem Potential verbunden. In dieser Studie korreliert die Expression von IMP3 mit TTF-1, Napsin A, histologischem Typ und klinischem Staging des Lungenkarzinoms. Wir waren daran interessiert, ob Fernmetastasen mit IMP3 Überexpression assoziiert sind, unabhängig von der histologischen Subtyp von Adenokarzinom.MethodeIn der retrospektiven Studie wurden die von 2006 bis 2009 im Klinischem Krankenhaus Split, Kroatien diagnostizerte Adenokarzinome der Lunge von 105 Patienten analysiert. Die klinischen Daten stammten aus der Abteilung für Pulmologie und im Falle des Todes vom Todesregister. Die Paraffinblöcke der primären Lungenbiopsate dieser Patienten wurden im Institut für Pathologie mit der indirekter Enzym - Immunohistochemie mittels Kombination der Antikörper gegen IMP3, Napsin A und TTF1 untersucht. 15 Fälle aus der Analyse aufgrund unzureichender Material ausgeschlossen. Es wurde eine statistische Untersuchung durchgeführt und Werte weniger als 0.05 P wurden als statistisch signifikant bezeichnet.ErgebnisseVon 90 Patienten mit Lungencarcinom waren 71 (78%) mänlich, durchschnittliches Alter war für Männer 61.5 Jahre (min-max 43–83) und 61 Jahre für Frauen (min-max 44–86). Pleurale Effusionen fand man in 15 Fällen (16.6%) und Fernmetastasen in 45 (50%) Fällen. Histologische Sybtypen waren: 2 lepidic Karzinome, 34 azinäre Karzinome, 2 papilläre und 52 solide Karzinome. IMP3 war exprimiert in 63 Fälle (70%). Positive IMP3 Expression war mit solidem Typ (P = 0.002) und negativer Napsin A Expression (P = 0.004) assoziert. Napsin A Expression war mit niedrigem Gradus (P = 0.031) und positiver TTF-1 Expression (P = 0.003) assoziert. Patienten mit IMP3 Überexpression öfter hatten Fernmetastasen als Patienten mit negativen IMP3, 55.5% versus 33.3% (P = 0.033). Non solide Subtyp mit IMP3 Überexpression entwickelten Fernmetastasen Meer häufiger als nicht festem Subtyp mit negativen IMP3, 72% versus 35% (P = 0.028).SchlussworteDie Expression von IMP3 ist mit negaativer Expression von Napsin A, solidem Subtyp und Metastasen verbunden und hat praktische predictive Werte in der pathologischen Diagnose des Adenokarzinoms der Lunge. Die Expression von IMP3 korreliert mit soliden Subtyp und mit Fernmetastasen unabhängig von histologische Subtyp Lungenadenokarzinom.

Morphological characteristics of placentas associated with idiopathic intrauterine growth retardation: a clinicopathologic study

OBJECTIVES To investigate histopathologic findings, placental diameters and characteristics of syncytial knots in the placentas from idiopathic intrauterine growth retardation (IUGR) pregnancies, and to compare them with a normal birth weight group. STUDY DESIGN Based on strict eligibility criteria, this prospective case-control study included 52 term placentas from idiopathic IUGR pregnancies and 69 term placentas from normal birth weight pregnancies. The study was carried out at the Clinical Hospital Centre, Split, where all placentas were collected and examined. For each placenta, diameters were measured and the following histopathologic findings were recorded: infarction, intervillous thrombosis, abruption, villous branching and maturation, chorioamnionitis, decidual vasculopathy and hemorrhagic endovasculitis for each placenta. In addition we assessed quantitative (number of syncytial knots and number of syncytial nuclei per syncytial knot) and qualitative (density and surface area) characteristics of syncytial knots in each placental sample. Statistical significance was tested using chi(2)-test, Students t-test and Mann-Whitney U-test. Statistical significance was set at P< or =0.05. RESULTS There was no difference in investigated histopathologic findings between idiopathic IUGR placentas and control group placentas. Placental diameters correlated significantly with neonatal birth weight (r=0.64; P<0.01); with higher birth weight there is an increase in placental diameters. Syncytial knots from idiopathic IUGR had significantly smaller surface area (Z=2.637; P=0.008) and higher density (Z=3.225; P=0.001) compared with the control group, while there is no difference in number of syncytial knots per individual villus, total number of syncytial knots in each placenta sample or number of syncytial nuclei per syncytial knot. CONCLUSIONS The investigated histopathologic findings in idiopathic IUGR placentas are incidental, with no higher frequency than in placentas from uncomplicated pregnancies, and should not be considered as possible causative factors for idiopathic IUGR. The demonstrated qualitative changes of syncytial knots in placentas associated with IUGR could represent a compensatory mechanism.

Trophoblast apoptosis in human term placentas from pregnancies complicated with idiopathic intrauterine growth retardation.

Objective. To investigate proliferative, apoptotic, and antiapoptotic activity of placental trophoblast in pregnancies complicated with idiopathic intrauterine growth retardation (IUGR). Methods. Study group included data and placentas from 52 normal singleton term pregnancies with idiopathic IUGR. Records and placentas from 69 singleton pregnancies with normal fetal growth served as a control group. IUGR was defined by birth weight less than 10th percentile of standard values. Children with congenital malformations and those born with the signs of hypoxia, laboratory or clinical signs of preeclampsia or infection, children born to anemic mothers and those born from pregnancies with an increased coagulation system activity were excluded. Results. There was no statistically significant difference in the cytotrophoblast proliferation index value (Z = 0.24; P = 0.553), trophoblast expression of the Bcl-2 antiapoptotic factor (Z = 0.47; P = 0.634), and trophoblast apoptotic index (Z = 0.51; P = 0.613) between the idiopathic IUGR and control group. Conclusion. The proliferative and apoptotic events in the trophoblast of placentas with idiopathic IUGR did not differ from physiologic ones. Study results suggest the IUGR syndrome to have no uniform etiology or even underlying pathophysiology that would determine the possible fetal risk and subsequent long-term consequences for fetal health and life. This imposes the need of a more precise definition and unambiguous distinction between the idiopathic and other forms of IUGR.

Difference in expression of collagen type I and matrix metalloproteinase-1 in uterosacral ligaments of women with and without pelvic organ prolapse

OBJECTIVE To compare the expression of collagen type I and matrix metalloproteinase-1 (MMP-1) in uterosacral ligament biopsies from women with and without pelvic organ prolapse (POP). STUDY DESIGN Uterosacral ligament biopsies were obtained from women with POP (n=46) and control subjects (n=49). Immunohistochemistry for collagen type I and MMP-1 was performed on formalin-fixed and paraffin-embedded sections. The two groups were matched for age, body mass index, parity and postmenopausal status. Statistical Package for the Social Sciences Version 13.0 was used for statistical analysis. RESULTS The expression of collagen type I (p=0.034) and MMP-1 (p=0.038) differed between women with POP and control subjects. There was increased expression of MMP-1 and decreased expression of collagen type 1 in uterosacral ligaments of women with POP compared with control subjects. CONCLUSIONS This difference indicates a possible relationship between POP and the immunohistochemical expression of collagen type I and MMP-1 in uterosacral ligaments.

Matrix metalloproteinases-1, -2 expression in uterosacral ligaments from women with pelvic organ prolapse

OBJECTIVE This study investigated matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-2 (MMP-2) immunohistochemical expression in uterosacral ligament biopsies from women with pelvic organ prolapse (POP), and controls with normal pelvic support. DESIGN Prospective observational experimental study. SETTING A tertiary Urogynecology Unit and Institute of Pathology, University Hospital Split, Croatia. POPULATION Women referred for hysterectomy for prolapse or benign gynecological disease. METHODS Eighty postmenopausal women were included in the study after Ethical Committee approval and informed consent. During surgery, uterosacral ligament biopsies were obtained from patients with POP (n=40) and women without evidence of pelvic floor weakening (n=40). Immunohistochemistry for MMP-1 and MMP-2 was performed on formaline fixed and paraffin embedded sections. Statistical evaluations were made by Student t-test or chi(2) test. MAIN OUTCOME MEASURES Immunohistochemical expression of MMP-1 and MMP-2 in the uterosacral ligaments of women with and without genital prolapse. RESULTS Forty women with POP and 40 controls without POP were included. The controls were matched to the women with POP in age, body mass index, parity and duration of postmenopausis. A significant increase in MMP-1 immunohistochemical expression was seen in uterosacral ligament tissue from women with POP (P=0.029). In contrast, there was no difference in immunohistochemical expression of MMP-2 between women with POP and those without (P=0.899). CONCLUSION These data suggest that MMP-1 may be marker of collagen degradation. Increased MMP-1 immunohistochemical expression in uterosacral ligaments is associated with urogenital prolapse.

Beneficial Effect of Nifedipine on Testicular Torsion-detorsion Injury in Rats

OBJECTIVE To investigate the effect of nifedipine on testicular torsion-detorsion injury. MATERIALS AND METHODS Twenty-four adult male Sprague-Dawley rats were randomly divided into 3 groups, each containing 8 rats. Rats in the control group underwent a sham operation of the left testis. In the torsion-detorsion (T/D) group, the left testis was twisted at 720° for 3 hours. After 3 hours of reperfusion, at the end of the experiment, the testes were removed. Rats in the treatment group received the same surgical procedure as the T/D group, but nifedipine was administered intraperitoneally (100 μg/kg) 30 minutes before the time of detorsion. RESULTS Unilateral testicular torsion-detorsion caused a significant increase in the malondialdehyde level and apoptosis and caused significant decreases in superoxide dismutase and glutathione peroxidase activities in ipsilateral testes. The rats treated with nifedipine had a significant decrease in malondialdehyde level and apoptosis and had significant increases in superoxide dismutase and glutathione peroxidase activities in ipsilateral testes compared with those of the T/D group. CONCLUSION These results suggest that biochemical and histological torsion-detorsion injury occurs in the ipsilateral testes after a 3-hour torsion and 3-hour detorsion and that administration of nifedipine before detorsion prevents ischemia/reperfusion cellular damage in the testicular tissue.

The significance of immunohistochemical expression of merlin, Ki-67, and p53 in meningiomas.

Meningiomas are one of the most common CNS tumors whose appearance is closely linked to NF2 gene product merlin. Tumor markers Ki-67 and p53 play established role in tumor progression which should be analyzed in close association with merlin expression. The aim of this study was to investigate the immunohistochemical expression of merlin in meningiomas, correlation with Ki-67 and p53, and to determine the association of these results with histologic grade and subtype. The histologic sections of 170 patients with totally resected meningiomas, between January 2000 and December 2010, were classified according to WHO, immunohistochemically stained for Ki-67, p53, and merlin, and analyzed using light microscope. Ki-67 median was 5.6 times higher in group of patients with negative merlin than in those with positive merlin (P=0.05). Statistically significant correlation of merlin with p53 was found (P<0.001). Merlin expression between 2 combined groups (meningothelial/secretory and fibroblastic/transitional) was statistically significant (P=0.002). By comparing merlin expression and p53 levels, statistically significant difference was found (P=0.017). In the group with positive merlin and negative p53 as well as positive merlin and low p53, meningothelial/secretory subtypes of meningiomas were more common. In combination of negative merlin and negative p53 as well as negative merlin and high p53, there were more meningiomas of fibroblastic/transitional subtype. There was no statistically significant correlation between merlin and tumor grade (P=0.420). There is undeniable influence of merlin on the development and the proliferative ability of meningioma subtypes. Significant role of p53 pathway was confirmed.

Cord blood cortisol level is lower in growth‐restricted newborns

Aim:  To establish the difference in plasma cortisol concentrations between newborns with intrauterine growth‐restricted (IUGR) and appropriate for gestational age (AGA) birthweights.