Genes involved in hormone metabolism and cellular response in human ovarian granulosa cells.

Endocrinal interactions are one of the most crucial regulatory mechanisms that maintain the state of homeostasis in humans. Processes such as oogenesis, folliculogenesis, menstruation and pregnancy remain under hormonal control. A key role in folliculogenesis is played by granulosa cells. Moreover, granulosa cells take part in corpus luteum formation after ovulation. Because of that, it is important to understand the ways in which the granulosa cells, associated with those processes, respond to hormonal stimulus. In the present study, a transcriptomic analysis of human granulosa cells (GCs) was carried out with the use of expression microarrays. The results were validated by RT-qPCR. The total RNA was isolated after 1st, 7th, 15th and 30th days of long-term primary cultures. The main focus of this work was placed on the genes belonging to "Response to estradiol", "Response to follicle-stimulating-hormone", "Cellular response to hormone stimulus", "Cellular hormone metabolic process" and "Hormone biosynthetic process" gene ontology groups. These groups of genes have been associated with GC hormone metabolism and cellular response to hormones. Eighty genes belonging to these groups were identified. Those that were members of more than one of the analyzed gene ontology groups, or exhibited unique expression patterns, were selected for further analysis. All of the selected genes were described, with their expression patterns detailed. In this manuscript, two gene expression patterns have been described. The first one showed large downregulation of genes in the later stages of culture, with the second one presenting upregulation of expression after day 1 of IVC. The present research was focused on six genes found to be the most important for steroidogenesis: STAR, POR, CYP11A1, ADM, GCLC, IL1B, as well as three genes of higher expression at the later stages of long-term in vitro culture: NR2F2, BMP4, COL1A1. The main goal of the presented study was to select genes involved in response to hormonal stimulus and hormone metabolism in GC long-term in vitro culture.

Overview of the different methods used in the primary culture of oral mucosa cells.

The culture of primary cells in vitro has enabled to gain knowledge in the field of cell biology, disease mechanisms and to offer great potential in drug testing. To date, two main techniques of isolating and culturing oral mucosal cells, the direct explant method and the enzymatic method, dominate the literature and practice. In the present study, both techniques are discussed in detail, comparing the advantages and disadvantages of the two approaches in setting up a primary culture of oral mucosal cell. The direct explant technique is well-established and has been commonly used for the past 20-30 years. Although the method of setting up the cultures did not show much variations in the methodology described by authors, the culturing conditions varied according to the aims of the projects.

Insight into "nuclear-cytoplasmic shuttling" as a developmental and differentiational capability of cells in primary culture models.

Shuttling proteins are molecules that can facilitate transport through the nuclear envelope. A very large number of proteins are involved in this process that includes nuclear pore buildup, signal, receptor and enzyme proteins. There are many examples of proteins whose biological activity depends on nucleocytoplasmic transport. Very often they are largely responsible for the proper occurrence of cell division, maturation, development and differentiation. Thanks to the well mastered methods of in vitro cell culture, it is possible to trace the levels of protein expression and their distribution in cells. Advanced molecular techniques allow for precise determination of their displacement in time. Several studies are still being carried out, using primary cultures, to identify the factors that determine the maturation, development and differentiation of cells. In understanding of the detailed mechanisms controlling cell life, the key is not the level of expression of a specific protein, but its distribution in individual cellular compartments.

Transcriptomic profile of cell cycle progression genes in human ovarian granulosa cells.

The ovarian granulosa cells (GCs) that form the structure of follicle undergo substantial modification during the various stages of human folliculogenesis. These modifications include morphological changes, accompanied by differential expression of genes, encoding proteins which are mainly involved in cell growth, proliferation and differentiation. Recent data bring a new insight into the aspects of GCs' stem-like specificity and plasticity, enabling their prolonged proliferation and differentiation into other cell types. This manuscript focuses attention on emerging alterations during GC cell cycle - a series of biochemical and biophysical changes within the cell. Human GCs were collected from follicles of women set to undergo intracytoplasmic sperm injection procedure, as a part of remnant follicular fluid. The cells were primarily cultured for 30 days. Throughout this time, we observed the prominent change in cell morphology from epithelial-like to fibroblast-like, suggesting differentiation to other cell types. Additionally, at days 1, 7, 15 and 30, the RNA was isolated for molecular assays. Using Affymetrix® Human Genome U219 Array, we found 2579 human transcripts that were differentially expressed in GCs. From these genes, we extracted 582 Gene Ontology Biological Process (GO BP) Terms and 45 KEGG pathways, among which we investigated transcripts belonging to four GO BPs associated with cell proliferation: "cell cycle phase transition", "G1/S phase transition", G2/M phase transition" and "cell cycle checkpoint". Microarray results were validated by RT-qPCR. Increased expression of all the genes studied indicated that increase in GC proliferation during long-term in vitro culture is orchestrated by the up-regulation of genes related to cell cycle control. Furthermore, observed changes in cell morphology may be regulated by a presented set of genes, leading to the induction of pathways specific for stemness plasticity and transdifferentiation in vitro.

[Still's disease as an example of fever of unknown origin: diagnostic and therapeutic difficulties]. / Choroba Stilla przyczyna goraczki niejasnego pochodzenia: trudnosci diagnostyczno-róznicowe i lecznicze.

A case of systemic form of juvenile chronic arthritis (Still's disease) is presented. Difficulties in confirmation of diagnosis are described. After nonsteroidal-antiinflammatory drugs and cytostatics administration very good therapeutical effect was achieved.

[LGM inferior vena cava filters--follow up 50 patients]. / Filtry LGM zakladane do zyly glównej dolnej--obserwacja 50 chorych.

In the Department of Medicine at the Institute of Tuberculosis and Lung Diseases 50 LGM inferior vena cava filters have been inserted since 1993. Indications for filters placement were as follows: recurrent pulmonary embolism (PE) despite anticoagulation-16 patients (pts), severe bleeding complications of thrombolytic or anticoagulant therapy-9 pts, contraindications for thrombolytic and/or anticoagulant treatment-3 pts, massive PE-6 pts, chronic thromboembolic-major vessel pulmonary hypertension (CTEPH)-18 pts, extensive deep vein thrombosis of lower limbs or vena cava inferior in patients with urgent indications for surgery-10 pts. In every patient diagnostic procedures were performed after 1, 3, 6, 12, 24 and 36 months of follow-up period. Only one non-fatal episode of recurrent PE was documented. Other complications were rare and insignificant. The LGM inferior vena cava filters are effective and safe in such selectively chosen group of patients.

[Clinically silent pulmonary embolism in patients with proximal deep venous thrombosis]. / Klinicznie bezobjawowy zator tetnicy plucnej u chorych z zakrzepica odcinka proksymalnego zyl glebokich konczyn dolnych.

In 18 patients with proximal deep venous thrombosis (PDVT) confirmed by phlebography, no symptoms and signs of pulmonary embolism (PE) were observed. All patients were treated with nadroparin. During first 6 days of treatment in all patients perfusion lung scans were performed. 8 patients (44.4%) of all group developed lung scans positive for PE (silent PE). Period of successful treatment of PDVT was 10 days. No evidence of recurrent PE were observed during the period of treatment. We conclude that: 1. Frequency of silent PE in patients with PDVT is very high-lack of symptoms and signs of PE does not exclude the presence of PE in this group of patients. 2. In all patients with PDVT perfusion lung scan should be performed even in cases with no symptoms and signs of PE. 3. Low molecular weight heparins administered subcutaneously are effective in treatment either silent PE or PDVT.

[Low doses of rtPA administered as a bolus in treatment of clinically acute massive pulmonary embolism]. / Male dawki rtPA podawane w bolusach w leczeniu klinicznie masywnego zatoru tetnicy plucnej.

12 patients (7 male and 5 female) with confirmed pulmonary embolism (PE) with: angiography-5 cases, conventional contrast-enhanced CT-2 cases, echocardiography-2 cases, autopsy-3 cases were diagnosed as clinically acute PE. Criteria of clinically acute PE were: cardiac arrest-1 case-2 cases, shock-1 case, acute cor pulmonale-9 cases and acute cor pulmonale with shock. All patients were treated with heparin, administered with therapeutic prolongation of aPTT. Clinically acute PE (if possible confirmed with angiography, TC and/or echocardiography) was treated with rtPA administered in 10 minutes lasting bolus in doses 0.6-0.8 mg per kg of body weight (50 mg of rtPA during 10 minutes administered into peripheral veins). In 9 patients with pulmonary hypertension, significant decrease of tricuspidal gradient (measured echocardiographically during several hours after administration of rtPA) was documented. Improvement in PaO2, SaO2 and decrease of heart rate and respiratory rate were also achieved. No serious bleeding complications were observed after mentioned treatment. Control investigations (conventional contrast-enhanced CT and spiral CT) performed several days after rtPA administration revealed thrombus in pulmonary artery. We conclude: I rtPA administered in bolus simultaneously with heparin significantly decreased pulmonaryhypertension; rtPA administered simultaneously with heparin is safe method of treatment of PE; hemodynamic improvement after administration of rtPA is not univocal with full fibrynolitic effect.

[Evaluation of intrapericardial cisplatin administration in cases of recurrent malignant pericardial effusion and cardiac tamponade]. / Ocena skutecznosci leczniczej wewnatrzosierdziowego zastosowania cysplatyny w przypadkach nawracajacych wysieków nowotworowych i tamponady serca.

30 consecutive patients with large malignant pericardial effusion (MPE) entered this prospective study. After pericardiocentesis and insertion of a polyurethane catheter, pericardial fluid was drained. Malignant etiology of pericardial fluid was confirmed by cytological examination. After confirmation of MPE cisplatin (10 mg in 20 ml normal saline) was instilled over 5 minutes during 5 consecutive days directly into pericardial space. If fluid reaccumulation occurred the courses were repeated every 3 weeks. Treatment was considered successful if the patient with malignant effusion survived 30 days without recurrence of symptoms of large pericardial effusion and other interventions directed to the pericardium were required. Positive effect of intrapericardial treatment with cisplatin was achieved in 18 cases (60%). Mean period of response was 3, 7 months (+/-6). Cisplatin administered directly into pericardial space is effective and safe method of treatment of recurrent MPE. Sclerosis of the pericardial space is rare complication connected with CP. Positive effect of CP can depend on improvement of lymphatic drainage from heart. CP seems to be method of choice in intal intrapericardial treatment in patients with malignant cardiac tamponade and recurrent MPE in course of lung cancer.

[Very low doses of tissue plasminogen activator in treatment of acute massive pulmonary embolism]. / Bardzo male dawki tkankowego aktywatora plazminogenu w leczeniu masywnego zatoru tetnicy plucnej.

57 years old woman with clinically acute massive pulmonary embolism confirmed by CT enhanced by contrast administration was treated with very low dose of rtPA (0.33 mg/kg) simultaneously with constant infusion of heparin (with therapeutic prolongation of aPTT). Excellent clinical effect and decrease of SPAP were achieved.

[Coronary artery vasospasm as a cause of chest pain in a patient with small cell lung cancer]. / Kurcz tetnic wiencowych jako przyczyna bólu w klatce piersiowej u chorego na drobnokomórkowego raka pluca.

Case of 67 years old man with small cell lung cancer and coronary artery spasm has been presented. After administration calcium channel blockers and nitroglycerin very good therapeutical effect was achieved.

[Prophylactic use of the LGM filter in a patient with a recurrent clinically acute massive pulmonary embolism complicated by pericarditis--case report]. / Profilaktyczne zastosowanie filtru LGM u chorego w nawracajaa klinicznie masywna zatorowoscia plucna powiklana zapaleniem osierdzia--opis przypadku.

Case of recurrent, clinically acute massive pulmonary embolism treated with rtPA (administered 0.6 mg/kg, during 10 minutes simultaneously with heparin) is presented. Minimal clinical improvement was observed after mentioned procedure. Good clinical response was achieved after LGM filter insertion into vena cava inferior. Clinical course was complicated by Dressler-like syndrome successfully treated with steroids. Problems of massive pulmonary embolism, vena cava filter prophylaxis and pericardial complication of pulmonary embolism are discussed.