Aktywność PARP-1 w prawidłowym i nowotworowym endometrium w relacji do liczby miejsc apurynowych/ pirymidynowych

Objectives: Poly (ADP-ribose) polymerase (PARP-1) is involved in the processes of DNA repair, contributing to the maintenance of genomic stability. Recent data suggest that polymerase is involved in the development of endometrial adenocarcinomas and more advanced tumors displaying lowest enzyme protein expression. Data on PARP-1 activity regarding carcinogenesis in human endometrium are scarce. That was the reason why the authors of the present work wished to investigate the enzyme activity in human uterine hormone-dependent cancer and to compare the results with those obtained for normal endometrial tissue. The next aim was to check whether enzyme activity in normal and cancerous endometrium depends on the number of AP sites, which are widely known as oxidative stress DNA damage markers and PARP-1 activity stimulators. Material and methods: Universal Colorimetric PARP Assay Kit was used to estimate the enzyme activity in units/mg protein. Apurinic sites/105 base pairs (bp) were measured by Oxidative DNA Damage Kit Quantitative. Results were calculated for 47 endometrial samples and 15 uterine adenocarcinomas specimens. Finally, the PARP-1 activity was analyzed for histological and some clinical features of neoplasms. Results and conclusions: 1. no differences in PARP-1 activity were found in non-cancerous types of human endometrium; 2. mean enzyme activity was lower in sporadic endometrial cancers than in noncancerous endometrial specimens (2.89±0.55 vs 6.39±0.06; pStreszczenie Cel pracy: badanie aktywności polimerazy poly(ADP-ryboza) (PARP-1) w prawidłowej i w nowotworowo zmienionej błonie śluzowej macicy w relacji do liczby miejsc apurynowych/apirymidynowych (AP), uznanych za aktywatora enzymu. Materiał i metody: badania wykonano w utkaniu 15 gruczolakoraków endometrium oraz 47 fragmentach prawidłowej błony śluzowej macicy. Aktywność PARP oznaczano przy użyciu Universal Colorimetric PARP Assay Kit (Trevigen) i przeliczano na miligram białka. Liczbę miejsc AP kwantyfikowano w 105 par zasad wykorzystując Oxidative DNA Damage Kit Quantitative (Kamiya). Aktywność polimerazy w utkaniu nowotworowym analizowano w relacji do niektórych histologiczno-klinicznych cech guzów gruczołowych. Wyniki i wnioski: 1. nie stwierdzono różnic aktywności PARP-1 w relacji do typu histologicznego nowotworowo niezmienionej błony śluzowej macicy; 2. średnia aktywność enzymu była niższa w utkaniu gruczolakoraków niż w endometrium nienowotworowym (2,89±0,55 vs. 6,39±0,06;

8-oxo-7,8-dihydroguanine level – the DNA oxidative stress marker - recognized by fluorescence image analysis in sporadic uterine adenocarcinomas in women

Objectives: In the case of carcinogenesis in human endometrium no information exists on tissue concentration of 8-oxo-7,8-dihydroguanine, the DNA oxidative stress marker. This was the main reason to undertake the investigation of this DNA modification in human uterine estrogen-dependent tissue cancers. Material and Methods: In order to estimate the level of oxidative damage, 8-oxo-7,8-dihydroguanine was determined directly in cells of tissue microscope slides using OxyDNA Assay Kit, Fluorometric. Cells were investigated under confocal microscope. Images of individual cells were captured by computer-interfaced digital photography and analyzed for fluorescence intensities (continuous inverted 8-bit gray-scale = 0 [black]-255 [white]). Fluorescence scores were calculated for each of 13 normal endometrial samples and 31 uterine adenocarcinoma specimens. Finally, the level of the oxidative stress marker was also analyzed according to histological and clinical features of the neoplasms. Results: The obtained data revealed that: 8-oxo-7,8-dihydroguanine levels were higher in uterine adenocarcinomas than in normal endometrial samples (48,32 vs. 38,64;

Czy globalna metylacja DNA w gruczolakorakach endometrium u kobiet jest zależna od histologicznego i klinicznego zaawansowania nowotworu?

Aim: to find out the relationship, if any, between the extent of the overall genomic DNA methylation, and clinical and pathological features of the sporadic endometrial adenocarcinomas in women. Material: genomic DNA was isolated from 44 primary uterine cancer tissue specimens. There were eight G1, 24 G2 and twelve G3 tumors. Methods: m5dC level was estimated after enzymatic digestion of DNA into nucleotides, 32P-postlabelling, twodimensional thin-layer chromatography on cellulose plates and phosphorobioimaging. The overall m5dC of the uterine cancer DNAs expressed as a ratio: (pm5dC/pm5dC+pdC) x 100% was compared to results obtained for parallel investigated but published earlier normal human endometrium DNAs. Results: mean total cancer DNA methylation (3.48±0.46%) was significantly higher than that of the normal proliferative endometrium (2.94±0,4%, p=0.003) and lower than that of the secretory endometrium DNAs (3.75±0,47%, p=0.03). Among all endometrial cancer DNAs six were found to be hypomethylated, eight were hypermethylated, whereas the remaining 30 had m5dC within range of normal endometrium. Total DNA methylation was significantly higher in poorly differentiated (G3) than in lower grade neoplasms (3.94±0.46 vs. 3.3 ±0.32 %, p=0.025). Lower levels of DNA methylation seemed to be associated with diminished tumor invasiveness. Conclusions: our results suggest that alterations in overall DNA methylation seem to be a result of neoplastic transformation and could therefore be used as a prognostic molecular marker of endometrial cancer.Streszczenie Cel pracy: poszukiwanie zależności między poziomem globalnej metylacji DNA, a niektórymi histologicznymi i klinicznymi cechami gruczolakoraków błony śluzowej macicy u kobiet. Materiał i metody: odsetek 5-metylocytozyny (m5dC) zbadano przy zastosowaniu metody 32P-postlabeling oraz dwukierunkowej chromatografii cienkowarstwowej (TLC 2-D) z kwantyfikacją radioaktywności nukleozasad przy zastosowaniu czytnika BAS 2000 w próbkach wysokooczyszczonych preparatów genomowego DNA wyizolowanego z 44 guzów (ośmiu w stopniu G1, 24 G2 i dwunastu G3). Wyliczony według wzoru: [(m5dC/ m5dC+dC) x 100] poziom zmodyfikowanej zasady korelowano z wiekiem chorych, odróżnicowaniem mikroskopowym guza oraz głębokością naciekania błony mięśniowej ścian macicy przez nowotwór. Poziom globalnej metylacji DNA w nowotworach przeanalizowano ponadto w relacji do uzyskanych równolegle, ale już opublikowanych, wyników badań stopnia metylacji w prawidłowym endometrium. Wyniki: średni poziom globalnej metylacji DNA w gruczolakorakach endometrium (3,48±0,46%) był wyższy niż zanotowano we wzrostowej (p= 0,003) i niższy niż stwierdzono w wydzielniczej błonie śluzowej macicy (p=0,03). Hipometylację stwierdzono w 6, podczas gdy hipermetylacja genomowego DNA wystąpiła w ośmiu nowotworach. Średnia, globalna metylacja guzów w stopniu G3 była wyższa niż odnotowano w pozostałych (3,94±0,46 vs 3,3±0.32 %, p=0,025). Stwierdzono narastanie poziomu zmodyfikowanego DNA w relacji do stopnia naciekania ścian macicy przez nowotwór. Wnioski: uzyskane wyniki wskazują, że zmiany metylacji cytozyny mogą być raczej skutkiem stopnia transformacji nowotworowej endometrium u kobiet, a nie przyczyną wystąpienia gruczolakoraka

Apurinic/apyrimidinic sites (AP) – stress oxidative marker – in normal and neoplastic human endometrium

Objectives: To evaluate apurinic/apyrimidinic sites (AP), one of the oxidative stress markers in normal and cancerous human endometrium and determine whether AP sites could be a molecular marker of endometrial cancer advancement. Material and methods: AP sites were investigated in DNAs of 33 endometrial cancer (EC) and 20 noncancerous endometrial samples using Oxidative DNA Damage Kit Quantitative (Kamiya Biomedical Company). Results: Mean AP sites in noncancerous endometrium was 6.0±1.21 per 105 base pair. In EC group the mean AP sites level was greater than estimated in the reference group containing proliferative, secretory and hyperplastic endometrial samples (

Vesicovaginal fistulas – current therapeutical strategy

Abstract Objectives: Clinical evaluation of Latzko operation in primary and recurrent vesicovaginal fistulas and an update on current therapeutical strategy. Material and methods: Evaluation of clinical efficacy has been performed in a group of 5 patients with confirmed vesicovaginal fistulas and in 1 woman with urethrovaginal fistula. Prior to the operation, four patients had undergone eight fistula operations in our department with unsuccessful outcome. Results: Fistula closure by Latzko operation has been achieved in 5 patients, including one patient who had surgery treatment repeated 3 months after the primary fistula operation. In total, additional Martius flap has been introduced twice. In patient, where fistula has been operated after 45 days since primary TVM and IVSO4 surgery, closure therapy failed. Conclusions: Surgical treatment by vaginal approach offers great therapeutical efficacy in the primary and recurrent vesicovaginal fistulas, even in cases when previous abdominal surgical treatment had failed

Conservative treatment of cervical pregnancy in women: a review of the literature with case report

Abstract Diagnosis of cervical pregnancy depends on ultrasonographic image of the gestational sac within the cervix, rather than histopathologic examination. Diagnosis of a cervical pregnancy may be confirmed if the placenta and the entire chorionic sac containing a live fetus are located below the internal os. Majority of patients with cervical pregnancies are young women who wish to stay fertile and have the possibility to bear children. Ultrasonography offers the advantage of early clinical diagnosis and easy fallow-up. Methotrexate (MTX) has become the treatment of choice in cervical pregnancy management, especially for the hemodynamically stable patients. In the following article we have reported a the successful management of a 7 weeks gestation cervical pregnancy treated with the combination of MTX, prostaglandin and suction curettage. Due to conservative treatment emergency hysterectomy was avoided

Zmiany w sposobie leczenia operacyjnego wysiłkowego nietrzymania moczu u kobiet – obserwacje ostatnich dwudziestu dwóch lat

Objectives: The aim of this study was to analyze the changing trends in surgical treatment of female urinary incontinence (UI). Material and methods: Medical records of all women admitted to II Department of Gynecology from 1985 to 2006 were analyzed in order to find out how the female SUI treatment changed over these years. Results: During analyzed time 36819 patients were hospitalized in our Department and 77.6% (28568) of them were operated because of various indications. The number of SUI surgeries among all hospitalized women steadily rose from 1.93% in 1985 to 10.96% in 2006 reaching maximum in 2005 (13.73%). Clinical effectiveness of SUI surgeries markedly improved from 35% for anterior colporrhaphy to almost 90 % for suburethral slings. Conclusions: Introduction into clinical practice modern suburethral slings improved clinical efficacy of SUI treatment. The percentage of women admitted and treated surgically because of SUI steadily increased over the last years.Cel pracy: Celem pracy było przeprowadzenie retrospektywnej analizy metod operacyjnego leczenia wysiłkowego nietrzymania moczu (WNM) u kobiet na przestrzeni ostatnich dwudziestu dwóch lat. Materiał i metody: Dane do analizy pozyskano z archiwum medycznego II Kliniki Ginekologii i na ich podstawie przedstawiono zmiany w operacyjnym leczeniu WNM u kobiet od 1985 do 2006 roku. Wyniki: W analizowanym okresie, w II Klinice hospitalizowanych było 36819 kobiet, z czego 77,6% (28568) było operowanych z powodu różnych wskazań. Liczba pacjentek operowanych z powodu WNM stale rosła od 1,93% w 1985r. do 10,96% w 2006r. ze szczytem w 2005r. (13,73%). Również skuteczność zabiegów znacznie wzrosła z 35% w przypadku różnych modyfikacji plastyki przedniej ściany pochwy do niemal 90% przy zastosowaniu taśm podcewkowych. Wnioski: Wprowadzenie nowoczesnych, małoinwazyjnych taśm podcewkowych znacząco zwiększyło efektywność operacyjnego leczenia WNM. Wydaje się to być jedną z przyczyn stałego wzrostu liczby kobiet decydujących się na tą metodę leczenia tej dolegliwości

Leczenie obrzęku limfatycznego miednicy – opis przypadku i przegląd piśmiennictwa

Pelvic lymphocoele is a result of systemic surgical lymph nodes removal leading to the accumulation of proteinrichinterstitial fluid in the extraperitoneal space. The presence of fluid arrested within the tissue accompanied bya decrease in the amount of oxygen and proteolytic activity evoke chronic infection. The excess of protein in thepresence of a bacterial infection additionally deteriorate lymphatic dysfunction. The most common bacteria causingthe infection are haemolytic streptococci. Moreover, the oedema makes tissue nutrition and healing of the wounddifficult. There are no randomised clinical trials devoted to specific lymphocoele treatment. The aim of the therapyis stabilization of lymphocyst, its decrease by needle aspiration and the improvement of the patient’s quality of lifeas well as the prevention and treatment of side effects. In the article we describe both surgical and pharmacologicaloptions of lymphocoele treatment with special attention paid to sclerotherapy