Impact of preimplantation genetic testing for aneuploidies (PGT-A) on first trimester biochemical markers - PAPP-A (placenta-associated plasma protein) and free ß-hCG (human chorionic gonadotropin).

OBJECTIVE: The objective of the study was to study the effect of preimplantation genetic testing for aneuploidies (PGT-A) performed at blastocyst stage on the levels of first trimester biomarkers. METHODS: This is an observational, collaborative, retrospective study. Seven hundred and twenty-eight patients were included in the study. Patients were with singleton pregnancies resulting from either natural conception (NC), or assisted reproductive techniques (ARTs) with PGT-A and frozen embryo transfer (FET) (ART/PGT-A/FET) or after ART without PGT-A and fresh ET (ART/no PGT-A/fresh ET) or FET (ART/no PGT-A/FET), who had first trimester combined screening test between 11 and 14 gestational weeks. They were stratified into four groups: group A (ART/PGT-A/FET) - 143 patients; group B (ART/no PGT-A/FET) - 100 patients; group C (ART/no PGT-A/fresh ET) - 346 patients, and group D (NC) - 139 patients. RESULTS: Statistically significant differences among the examined groups were observed for maternal age, BMI, ethnicity, and parity. The median placenta-associated plasma protein (PAPP-A) was lowest in the group with ART/PGT-A/FET and the highest result was obtained in the group with ART/no PGT-A/FET. Statistically significant difference in the median PAPP-A levels was identified among the examined groups (p = .0186). When a subgroup analysis was performed, a statistically significant difference was observed in the median PAPP-A between ART/PGT-A/FET group versus ART/no PGT-A/FET group (p = .01) and NC versus ART/no PGT-A/FET (p = .01). A similar trend toward statistical significance was noted when comparing NC versus ART/no PGT-A/fresh ET (p = .06). Multivariate analysis elucidated that when age is present in the model, the effect of any method of conception or testing for aneuploidy disappears. The other factors (BMI, ethnicity, and parity) do not influence the levels of PAPP-A. The lowest median free human chorionic gonadotropin (ß-HCG) was recorded in the NC group and the highest result was identified in the group with IVF/PGT-A/FET. No statistically significant difference was observed in the median concentration levels of free ß-hCG among the compared groups (p = .5789) and when subgroup analysis was performed (p>.05). The normality of the distribution of variables was analyzed by the Kolmogorov-Smirnov test and the median PAPP-A and free ßhCG concentration difference by the Wilcoxon rank-sum test with nonparametric ANOVA. CONCLUSIONS: Testing for aneuploidy (PGT-A) and the decision to transfer either fresh or cryopreserved embryos (ET) appear not to affect the levels of first trimester biochemical markers. The findings of the present study should be a baseline for future studies and could be used to improve the antenatal screening counseling for women with ART pregnancies and PGT-A.

Quality of life with talazoparib versus physician's choice of chemotherapy in patients with advanced breast cancer and germline BRCA1/2 mutation: patient-reported outcomes from the EMBRACA phase III trial.

Background: In the EMBRACA phase III trial, talazoparib (1 mg daily, orally) demonstrated a statistically significant improvement in PFS versus physician's choice of chemotherapy (PCT; capecitabine, eribulin, gemcitabine, or vinorelbine) in patients with HER2-negative advanced breast cancer carrying a germline BRCA1/2 mutation; we evaluated patient-reported outcomes (PROs). Patients and methods: Patients were randomized 2 : 1 to receive talazoparib or PCT. PROs were assessed at day 1 (baseline), the start of each treatment cycle (every 3 weeks), and at the end of treatment, using the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-30) and its breast cancer module, QLQ-BR23. Prespecified exploratory analyses included a longitudinal mixed-effect model comparing treatment arms and a time to definitive clinically meaningful deterioration (TTD) analysis carried out in the global health status/quality of life (GHS/QoL), and all functional and symptom scales from the EORTC QLQ-C30 and -BR23 questionnaires. Between-arm TTD comparisons were made using a stratified log-rank test and a Cox proportional hazards model. Results: Baseline scores were similar between arms. Statistically significant estimated overall improvement from baseline in GHS/QoL was seen for talazoparib compared with statistically significant deterioration for PCT {3.0 [95% confidence interval (CI) 1.2, 4.8] versus -5.4 [95% CI -8.8, -2.0]; between arms, P < 0.0001}. A statistically significant greater delay was observed in TTD in GHS/QoL, favoring talazoparib over PCT [hazard ratio, 0.38 (95% CI 0.26, 0.55; median, 24.3 versus 6.3 months, respectively; P < 0.0001)]. A statistically significant overall change and a statistically significant delay in TTD, all favoring talazoparib, were also observed in multiple functions and symptoms. Conclusion: Patients who received talazoparib had significant overall improvements and significant delay in TTD in multiple cancer-related and breast cancer-specific symptoms, functions, and GHS/QoL. ClinicalTrials.gov: NCT01945775.

[Mixed gonadal dysgenesis combined with gonadoblastoma].

Gonadal dysgenesis is defined by incomplete or defect forming of gonads, a result of disturbed process of migration of germ cells or and their correct organization in gonadal ridge. The combination of dysgenetic gonads and Y chromosome is a prerequisite for developing ovarian neoplasma--most frequent gonadoblastoma. We present a case of mixed gonadal dysgenesis at a patient with caryotype 46XY in combination with gonadoblastoma.

[Ovarian cancer with metastatic lesion in the vagina (foetus papiraceus)].

Ovarian cancer spreads primarily by intraperitoneal implantation of exfoliated cancer cells, by lymphatic dissemination, and by haematogenous spread. Very rarely it metastasizes to cervix, vulva and vagina; this type of metastases present a diagnostic challenge to the gynecologist and pathologist. We present a case of ovarian cancer with initial clinical manifestation-lesion of the vagina.

[Immunohistochemical profile of colorectal and ovarian carcinomas--examination with cytokeratin 7, cytokeratin 20, beta catenin and cDX 2].

INTRODUCTION: Metastases from colorectal adenocarcinomas can be histologically similar to serous, mucinous and endometrioid ovarian adenocarcinomas. The differentiation between primary and metastatic ovarian tumours is of great importance for the patients because of the different treatment and prognosis. AIM: The aim of the study was to determine whether the differences in the expression of Cytokeratin 7, Cytokeratin 20, Beta catenin and CDX2 can be used to distinguish the different types of carcinomas and their metastases. MATERIALS AND METHODS: The immunohistochemical expression of the listed above antibodies was examined retrospectively and prospectively in 38 colorectal adenocarcinomas (primary and metastatic) and 32 ovarian adenocarcinomas (primary and metastatic). The metastases in both types of adenocarcinomas are located in the peritoneum. RESULTS: The immunohistochemical expression was evaluated using a semi-quantitative method. The ovarian adenocarcinomas are mostly positive for Cytokeratin 7 (in 63%), while colorectal carcinomas are mostly positive for Cytokeratin 20 (in 73%). Regarding Beta catenin, in colorectal carcinomas the expression is mostly nuclear (in 65%) and in ovarian carcinomas mostly membrane (in 68%). In cases of uncertain expression of the markers mentioned above, CDX2 was used. Positive nuclear expression was observed only in intestinal tumours (in 86%). CONCLUSION: For differential diagnosis between ovarian and colorectal adenocarcinomas, the use of antibodies, determining the intestinal differentiation of the tumours like Cytokeratin 20, Beta catenin and CDX2 is recommended.

[Uterine carcinosarcoma with chondroid differentiation--immuhistochemical examination].

Uterine carcinosarcomas are highly aggressive malignant tumours, consisting of both epithelial and mesenchymal components. They represent 1-3% of the malignant uterine neoplasms. Their histogenesis is unclear. Because of their rarity and the very few clinical data available, the studies of potential therapeutic goals are scarce. We present clinical and pathohistological description of a rare case of a 76-year-old woman with a uterine carcinosarcoma with a chondroid differentiation, examined immunohistochemically with Cytokeratin, Vimentin and S-100 protein.

[CDX2--an immunochystochemistry marker of intestinal differentiation of adenocarcinomas].

CDX2 is a nuclear transcribing factor, important for the development and differentiation of the bowels. According to the recent publications, CDX2 expression is immunochystochemisry detector in the normal enterocytes of the bowels and is normally met in the most, but not at all colorectal carcinomas. Two homeostatic genes are detected at people CDX1 u CDX2. We present examination of CDX2 expression at 15 adenomas and 30 colorectal carcinomas. All the adenomas are positive for CDX2, 27 /90%/ of the adenocarcinomas show nuclear expression of CDX2.

[Differential diagnostic importance of immunochistochemistry applied at metastases of adenocarcinomas of the peritoneum].

Patients with metastatic carcinomas with unclear primary tumor site are often clinical and pathological problem. They are diagnosed in about 3-65 of the cases with solid tumors. In 60% of cases the reason is adenocarcinomas. The most frequent reason for peritoneal carcinomas at women is the ovarian cancer. In such cases we must exclude the probability of colorectal cancer. In fact there are not international standards for prove the origin of adenocarcinomas; different research groups use different criteria. We present the use of four antibodies--Cytokeratin 7 and 20, Beta cathenin and CDX2 and define their positiveness at metastases from different groups of adenocarcinomas in the peritoneum.

[Molecular genetics of ovarian cancer].

Ovarian cancer is the most lethal gynecologic neoplasia. The risk of ovarian cancer in Europe is 1 to 80. In 80-90% of cases, the clinical manifestation is in advanced stage. The 5 year survival is for III /IV stage 25%, but for I/II stage is 90%. In the present study we present the molecular and genetic changes of hereditary and sporadic ovarian cancer.

[Role of total inhibin in diagnosis and monitoring of ovarian cancer in postmenopausal women].

Ovarian cancer is the second and the most frequent fatal gynecological malignancy worldwide and the fourth reason for mortality of neoplasmas in Europe. The lifetime risk for ovarian cancer in Europe is approximately 1 in 80. One of the reasons for the high mortality rate is that in more than 70% of women with such cancer are diagnosed in advanced stage. After physiological menopause or after surgery, with the depletion of ovarian follicules (the granulose cells produce inhibins), the circulating serum inhibin A and B and the free alpha-subunit decrease low or non detectable levels. We present the significance of measuring total inhibin with new double ELISA method, as a diagnostic and prognostic factor in the sera of postmenopausal women with mucinous and granulose cells tumors. The serum total inhibin is a sensitive marker for diagnosis and monitoring of mucinous and granulose cells tumors as its levels decrease significantly after surgery and increase in case of recurrence. Total inhibin is useful marker with high specificity and sensitivity for these tumors; CA 125 for other varian cancer; the combination of both resulted in 95% detection of all ovarian cancers.

[A rare case of low grade endometrial stromal sarcoma of the uterus].

Endometrial stromal sarcoma (ESS) is a rare disease. Low-grade endometrial stromal sarcoma (LGESS) is characterized by proliferations composed of cells with endometrial stromal cell differentiation and typically show extensive worm-like vessel invasion. We present a case of a 38-year-old woman with complaints of abdominal pains and ultrasound findings of a tumour formation in the pelvis. The biopsy examination showed a low-grade endometrial stromal sarcoma (LGESS), with a primary location in the uterine corpus and a tumour infiltration in the myometrium and cervix of the uterus, with tumour emboli in the lymph vessels and tumour methastases in the peritoneum--FIGO III Stage. The patient underwent postoperative polychemotherapy and radiotherapy. The gynecological and ultrasound examination found no pathological changes in the pelvic region five years after the operation.

[Transcervical resection of the endometrium--an alternative of hysterectomy or not?].

Menorrhagia is frequent at women at reproductive age. Nowadays the standart for diagnosis of menorrhagia is hysteroscopy with histological verification. When there is no effect of the conservative treatment, an operative treatment is applied--hysterectomy or ablation of the endometrium. The aim of the present study is to present our experience with transcervical resection of the endometrium with classic abdominal hysterectomy as a therapeutic management for women with menorrhagia. Transcervical resection of the endometrium at selected patients (proved menorrhagia and length of the endometrium <10-12 sm) is a safe alternative of hysterectomy.

[Myasthenia gravis and pregnancy--a case report and review of the literature].

Myasthenia gravis is an acquired immune-mediated neuromuscular disorder, characterised by muscle weakness and fatigabillity of the voluntary muscles. It often affects women in the second and third decade of life with predilection of childbearing years (incidence in pregnancy--1:20,000). We present a case of a pregnant woman with myasthenia gravis and review the literature upon the problem - association of myasthenia gravis and pregnancy.

[Acute appendicitis and pregnancy].

Acute appendicitis remains the most common nonobstetric surgical diagnosis during pregnancy, requiring surgery. It is estimated that its frequency is 1 case per 1500 pregnancies. The delay in the diagnosis correlates with an increase in ruptured appendix and a dramatic elevation in fetal and maternal morbidity and mortality. In the study the authors discuss the clinical presentation, laboratory findings, operative treatment, pathologic diagnosis and the outcome of the disease of 17 cases with acute appendicitis in pregnancy, operated in the Clinic of Thoraco Abdominal Surgery for a ten years period of time. The early surgical treatment plays an important role for the succsessful outcome of the disease and reduces the complications for the mother and the fetus.

[Renal complications in severe acute pancreatitis]. / Bubrechni uslozhneniia pri tezhuk ostur pankreatit.

UNLABELLED: Except of the success in the diagnosis and treatment of acute pancreatitis, and at present this illness and its complications continue to be actual problem of urgent surgery. The object of our study is to establish the frequency of renal complications at patients with severe acute pancreatitis and to assess the most important risk factors of their development and mortality. MATERIAL AND METHODS: The object of our study is 198 patients with severe acute pancreatitis, 75 of whom have renal complications.

Tpl-2 induces apoptosis by promoting the assembly of protein complexes that contain caspase-9, the adapter protein Tvl-1, and procaspase-3.

The Tpl-2 proto-oncoprotein promotes cellular proliferation when overexpressed in a variety of tumor cell lines. Here, we present evidence that when overexpressed in immortalized non-transformed cells, Tpl-2 induces apoptosis by promoting the activation of caspase-3 via a caspase-9-dependent mechanism, and that apoptosis is enhanced when Tpl-2 is co-expressed with the newly identified ankyrin repeat protein Tvl-1. The activation of caspase-3 by caspase-9 is known to depend on the assembly of a multimolecular complex that includes Apaf-1 and caspase-9. Data presented here show that co-expression of Tpl-2 with Tvl-1 promotes the assembly of a complex that involves several proteins that bind Apaf-1 including Tvl-1, itself, Tpl-2 and phosphorylated procaspase-9. More important, procaspase-3, which under normal growth conditions is not associated with the complex, binds Tvl-1 conditionally in response to Tpl-2-generated apoptotic signals. The conditional association of procaspase-3 with Tvl-1 promotes the in vivo proteolytic maturation of procaspase-3 by caspase-9, a process casually linked to apoptosis.

Isolation of chromatin fragments released by apoptotic cells.

A two-step procedure for isolation of chromatin fragments released from apoptotic cells is described. The first step includes extraction of the chromatin fragments by a hypotonic treatment of isolated nuclei. The second step provides stabilization of the extracted chromatin fragments and their collection by a high-speed centrifugation. The procedure ensures not only a complete extraction and collection of the chromatin fragments but also intactness of their components. A number of control experiments confirmed this statement: they showed that the percentage of recovered fragmented DNA does not exceed 10% from the total DNA of apoptotic cells and that it is specific in both nature and chromosomal localization; the experiments reveal, moreover, that the protein components of the fragments including the well known histone and non-histone species possess also ability to support in in vitro conditions their specific phosphorylation. Developed for recovery of chromatin fragments from mouse erythroleukemia cells, spontaneously entering apoptosis, the procedure is practically applicable for all eukaryotic cell systems sharing programmable death.

Spontaneous apoptosis in mouse F4N-S erythroleukemia cells induces a nonrandom fragmentation of DNA.

This study characterizes the fragmented DNA of mouse erythroleukemia (MEL) cells spontaneously entering apoptosis. Fragmented DNA was isolated by introducing a novel procedure that ensured a complete extraction of the characteristic oligonucleosomal ladder. As the results show, less than 10% of DNA of apoptotic cells is fragmented in this form. The main conclusion from experiments to characterize the nature of fragmented DNA is that spontaneous apoptosis induces a nonrandom cleavage of genomic DNA. The Southern analysis performed with total apoptotic DNA revealed that it is strongly enriched in interspersed repetitive sequences. In situ hybridizations with such DNA showed further than in interphase nuclei these sequences flock together and form clusters spread throughout the whole nuclear area whereas in mitotic chromosomes they are located predominantly at their pericentromeric/peritelomeric ends. Partial cloning and sequencing reinforces the notion that the apoptotic DNA is representative for a heterochromatinic portion of the mouse genome. Support for such an unexpected conclusion is coming also from experiments indicating that this DNA is heavily methylated and poorly transcribed.

Interphase chromosomes of Friend-S cells are attached to the matrix structures through the centromeric/telomeric regions.

DNA of the attachment sites of Friend erythroleukemia cells, isolated according to the conventional procedure, represents short, nuclease-resistant fragments with sizes below 400 bp, belonging to the class of mouse satellite. A number of experiments have indicated that their unusual resistance is due to complexing with RNA. By various approaches, it was confirmed that similar fragments might be recovered from total DNA following extensive digestion with DNase I. In situ hybridizations revealed further that at mitosis the sequences of the attachment sites are located at the centromeric/telomeric regions of the chromosomes, while at interphase they are redistributed into 9-13 well-defined clusters spread throughout the entire nuclear area. Parallel biochemical and electronmicroscopic studies have clarified, moreover, that the all three compartments of the matrix harbor such sequences. Thus, it appears that the attachment sites described function only at interphase, anchoring the both ends of each interphase chromosome to the matrix structures.

Spatial and structural segregation of the transcribed and nontranscribed alleles of c-myc in Namalva-S cells.

By using various approaches we received evidence that, in Namalva-S cells carrying a t(8;14) translocation and highly expressing c-myc, the two alleles of the gene are spatially and structurally segregated. Spatial segregation of the alleles was observed in all nuclei analyzed by in situ hybridization technique. Their structural segregation, i.e., association with different intranuclear structures, was confirmed in a number of experiments. When high-salt extracted nuclei were digested with EcoRI, which is known to produce fragments containing the entire c-myc locus, the sequences of the gene were found separated between the pellet, containing sequences firmly associated with the heavier matrix structures, and the supernatant, containing sequences from the free length of the DNA loops. Southern hybridization performed with a probe representative for the constant region of the human IgH locus revealed that this fractionation in fact segregates the reorganized from the normal allele of c-myc. Run-on experiments carried out with two fractions, topologically equivalent to the above P and S but isolated as intact chromatin structures, indicated that the allele associated with nuclear matrix is actively transcribed, while that located in the free length of the chromatin loops is practically nontranscribed. Studies on the chromatin organization of transcribed and nontranscribed alleles revealed the existence in them of two alternative chromatin structures. Control experiments with beta-globin gene, performed with cells constitutively nontranscribing or actively transcribing this gene, confirmed our conclusions about the spatial segregation of the two alleles and clarified that their structural segregation occurs when the gene is activated for transcription.