Animal models of liver diseases and their application in experimental surgery.

Both acute and chronic liver diseases are frequent and potentially lethal conditions. Development of new therapeutic strategies and drugs depends on understanding of liver injury pathogenesis and progression, which can be studied on suitable animal models. Due to the complexity of liver injury, the understanding of underlying mechanisms of liver diseases and their treatment has been limited by the lack of satisfactory animal models. SO far, a wide variety of animals has been used to mimic human liver disease, however, none of the models include all its clinical aspects seen in humans. Rodents, namely rats and mice, represent the largest group of liver disease models despite their limited resemblance to human. On the other hand, large animal models like pigs, previously used mostly in acute liver failure modeling, are now playing an important role in studying various acute and chronic liver diseases. Although significant progress has been made, the research in hepatology should continue to establish animal models anatomically and physiologically as close to human as possible to allow for translation of the experimental results to human medicine. This review presents various approaches to the study of acute and chronic liver diseases in animal models, with special emphasis on large animal models and their role in experimental surgery.

Relationship between follicular volume and oocyte competence, blastocyst development and live-birth rate: optimal follicle size for oocyte retrieval.

OBJECTIVE: To analyze oocyte competence in gonadotropin-releasing hormone agonist (GnRHa) stimulation cycles with regard to maturity, fertilization and blastocyst rate, as well as clinical outcome (pregnancy and live-birth rate), in relation to follicular volume, measured by three-dimensional transvaginal sonography (3D-TVS), and follicular fluid composition. METHODS: This was a prospective single-center study conducted between June 2012 and June 2014, including 118 ovum pick-ups with subsequent embryo transfer. Ovarian stimulation was performed using the GnRHa long protocol. Of 1493 follicles aspirated individually, follicular volume was evaluated successfully in 1236 using automated 3D-TVS during oocyte retrieval. Oocyte maturity and blastocyst development were tracked according to follicular volume. Intrafollicular concentrations of estradiol, testosterone, progesterone, luteinizing hormone, follicle-stimulating hormone and granulocyte-colony stimulating factor were quantified by immunoassay. Clinical outcome, in terms of implantation rate, (clinical) pregnancy rate, miscarriage and live-birth rate (LBR), was evaluated. RESULTS: Follicles were categorized, according to their volume, into three arbitrary groups, which included 196 small (8-12 mm/0.3-0.9 mL), 772 medium (13-23 mm/1-6 mL) and 268 large (≥ 24 mm/> 6 mL) follicles. Although oocyte recovery rate was significantly lower in small follicles compared with medium and large ones (63.8% vs 76.6% and 81.3%, respectively; P < 0.001), similar fertilization rates (85.1% vs 75.3% and 81.4%, respectively) and blastocyst rates (40.5% vs 40.6% and 37.2%, respectively) per mature metaphase II oocyte were observed. A trend towards higher LBR after transfer of blastocysts derived from small (< 1 mL) follicles compared with medium (1-6 mL) or large (> 6 mL) follicles (54.5% vs 42.0%, and 41.7%, respectively) was observed. No predictive value of follicular fluid biomarkers was identified. CONCLUSIONS: Our data indicate that the optimal follicular volume for a high yield of good quality blastocysts with good potential to lead to a live birth is 13-23 mm/1-6 mL. However, oocytes derived from small follicles (8-12 mm/0.3-0.9 mL) still have the capacity for normal development and subsequent delivery of healthy children, suggesting that aspiration of these follicles should be encouraged as this would increase the total number of blastocysts retrieved per stimulation. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

[Epithelial-mesenchymal Transition in Tumor Tissue and Its Role for Metastatic Spread of Cancer]. / Epitelo-mezenchymální tranzice v nádorové tkáni a její role pri metastatickém sírení karcinomu.

BACKGROUND: Metastasis, recurrence, and resistance to chemotherapy are leading causes of the majority of cancer-related mortality worldwide. The process of metastasis can be artificially divided into a series of sequential, highly organized, and organ-specific steps. The underlying mechanisms are still poorly understood, but are believed to be mediated by epithelial-mesenchymal transition (EMT). First described in embryogenesis, EMT is a cellular reprogramming process in which epithelial cells acquire a mesenchymal phenotype. During this transformation, epithelial cells lose their shape, epithelial markers, and ability to grow in colonies. They acquire a spindle-shaped morphology and exhibit more motile and invasive behavior. These phenotypic changes are associated with modifications in different interconnected protein and gene families, such as transcription factors, cadherins, catenins, matrix metalloproteases, and growth receptors. EMT has been observed in many cancers, such as breast, ovarian, colon, and esophageal cancers, and is associated with poor prognosis and metastasis. Also, resistance to cytotoxic treatments is associated with reactivation of embryonic programs. Understanding this process is necessary to provide a better understanding of cancer progression and could lead to the development of new therapeutic or prognostic strategies for the treatment of cancer. CONCLUSION: This article summarizes the known molecular pathways involved in EMT in cancer.Key words: epithelial-mesenchymal transition - carcinoma - metastasisThe authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 24. 6. 2016Accepted: 14. 11. 2016.

Porcine liver vascular bed in Biodur E20 corrosion casts.

BACKGROUND: Pigs are frequently used as animal models in experimental medicine. To identify processes of vascular development or regression, vascular elements must be recognised and quantified in a three-dimensional (3D) arrangement. Vascular corrosion casts enable the creation of 3D replicas of vascular trees. The aim of our study was to identify suitable casting media and optimise the protocol for porcine liver vascular corrosion casting. MATERIALS AND METHODS: Mercox II® (Ladd Research, Williston, Vermont, USA) and Biodur E20® Plus (Biodur Products, Heidelberg, Germany) were tested in 4 porcine livers. The resins (volume approximately 700 mL) were injected via the portal vein. Corrosion casts were examined by macro-computed tomography, micro-computed tomography and scanning electron microscopy. RESULTS: For hepatectomies, the operating protocol was optimised to avoid gas or blood clot embolisation. We present a protocol for porcine liver vascular bed casting based on corrosion specimens prepared using Biodur E20® epoxy resin. CONCLUSIONS: Only Biodur E20®Plus appeared to be suitable for high-volume vascular corrosion casting due to its optimal permeability, sufficient processing time and minimum fragility. Biodur E20® Plus is slightly elastic, radio-opaque and alcohol-resistant. These properties make this acrylic resin suitable for not only vascular research but also teaching purposes.

[Czech registry of complications in gynecological laparoscopy - the on-line data collection]. / Národní registr komplikací gynekologické laparoskopie CR -první zkusenosti s elektronickým sberem dat a vize do budoucna.

OBJECTIVE: To survey the performance of laparoscopically assisted vaginal hysterectomy (LAVH) and other gynecologic laparoscopic operations performed in the Czech Republic. DESIGN: Retrospective study (Canadian Task Force classification II-3). SETTING: Departments of gynecology in the Czech Republic. METHODS: Two paper questionnaires were sent to 120 clinics performing laparoscopic gynecologic operations in the past, the data have been collected via web application since 2013. RESULTS: Process of transition from paper questionnaires to electronic forms was realized without complications. Number of clinics that responded was the highest during 5 last years. Number of reported gynecologic laparoscopic operations in 2013 was the second highest since the establishment of the Registry. CONCLUSION: Unchanged trend of decrease in number of abdominal hysterectomy. Latest data shows the possibility of vaginal hysterectomy renaissance.

[Sperm mitochondrial function in men with normozoospermia and asthenozoospermia]. / Funkce mitochondrií ve spermii u muzu s normozoospermií a astenozoospermií

OBJECTIVE: One of causes of male infertility is reduced sperm motility. It turns out that the reduced efficiency of the mitochondrial respiratory activity may play a role in the development of this disorder. The aim of our study was to comprehensively determine mitochondrial respiratory activity of sperm with normal and reduced motility. DESIGN: Prospective study. SETTING: Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague; Department of Physiology, Faculty of Medicine in Pilsen, Charles University in Prague; Institute of Reproductive Medicine and Endocrinology, IVF Centers Prof. Zech, Plzen. METHODS: Ejaculates of 14 men were obtained from IVF Center Prof. Zech, Pilsen. According to the World Health Organization classification, samples were divided into normozoospermatic (n = 7) and asthenozoospermatic(n = 7) groups. Respiratory activity of sperm was measured on two-chamber oxygraph Oroboros. RESULTS: In asthenozoospermatic samples, significantly reduced activity of complex I (p = 0.007) and increased respiration after application of ATP-synthase inhibitor oligomycin (showing increased uncoupled oxidation and phosphorylation, p = 0.046) were found. Inhibition of complex I by rotenone showed that complex I contribution to the total capacity of oxidative phosphorylation of healthy sperm was relatively lower than it is typical for somatic cells. CONCLUSION: In our study, we measured mitochondrial respiratory activity of human sperm, permeabilized by digitonin, by high-resolution oxygraphy, which allows the determination of oxygen consumption from the smallest possible number of germ cells. The study results confirm reduced activity of complex I in asthenozoospermatics and suggest that increased leakage of protons from the mitochondrial matrix, which leads to reduced efficiency of phosphorylating process, could participate in the reduced sperm motility. Better characterization of male germ cells, either completely healthy or with affected motility, will help us to understand better the physiological process of fertilization and also to choose the most viable sperm for infertility treatment by methods of assisted reproduction.

[Effect of overweight on the outcome of assisted reproductive technology in infertile women]. / Vliv nadváhy na úspesnost technik asistované reprodukce u neplodných zen.

OBJECTIVE: Obesity and overweight negatively affect the ability of women to conceive naturally, contributes to the increased incidence of obstetric complications during pregnancy and affect the outcome of assisted reproduction techniques (ART). The aim of our study was to compare the results of treatment of infertilityin a group of infertile women undergoing ART, depending on the values of BMI and changes in levels of selected hormones and markers of oxidative stress in follicular fluid. DESIGN: Retrospective comparative study. SETTING: Faculty of Medicine in Pilsen, Charles University in Prague; Institute of Reproductive Medicine and Endocrinology, IVF Centers - Prof. Zech, Plzen. METHODS: The studied group consisted of 44 women (mean age of 31.9 years, SD = 4.35) treated for infertility at the Institute of Reproductive Medicine and Endocrinology - IVF Centers Prof. Zech. Women were divided into 2 groups according to BMI (37 women had normal BMI, 7 women were overweight). Prolactin, free T3 and T4 hormone, homocysteine, malondialdehyde, glutathione peroxidase, total antioxidant capacity and total protein were analyzed in the follicular fluid both groups. Only blood free samples were studied after pooling of all FF samples of each patient. RESULTS: We observed significantly lower levels of glutathion peroxidase in the group of overweight women(p = 0.0044). The pregnancy success rate with women with normal BMI and overweight women did not differ significantly from each other (p = 0.4430). CONCLUSION: Our study did not confirm the negative effect of obesity on the results of treatment of infertility, specifical-ly pregnancy rate in a group of infertile women undergoing treatment with assisted reproduction techniques.

[Analysis of point mutations in interleukin-11 gene in the population of infertile patients and fertile control women]. / Vysetrení bodových mutací v genu pro interleukin-11 v populaci neplodných zen a plodných kontrol.

OBJECTIVE: The interleukin-11 (IL-11) is one of the most important signaling factors in the process of decidua-lization and embryo-maternal cross talk during the embryo implantation. We investigated the prevalence of the IL-11 gene point variants in the population of infertile women and fertile control subjects. DESIGN: A cohort study. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine and University Hospital of Charles University, Pilsen. SUBJECTS AND METHODS: The population to screen consisted of 354 infertile women. The control population was comprised of 195 healthy fertile subjects. For the mutational analysis, the temperature gradient gel electrophoresis (TGGE) and direct sequencing had been used. The groups of fertile controls and infertile patients were compared for statistically significant difference using the Fisher´s 2 by 2 Exact test. RESULTS: Thirteen potentially functional IL-11 gene variants, the G to A transversions at the position 3651 (G3651A) leading to the arginin to histidin exchange on the position 113 (R113H) were detected in the group of infertile women. No mutations were identified in the control group, which means that the frequency of functionally relevant mutations of the IL-11 gene in infertile women is significantly enhanced in comparison with controls (p = 0.01, Fisher´s 2 by 2 Exact test). CONCLUSION: The results prove the presence of the potentially functionally relevant IL-11 gene variants in the population of infertile women. The presence of variants does not have to lead directly to decrease fertility but may contribute to the dysbalance of cytokine milieu which is needed for the decidualization as well as subsequent embryo implantation. Even thought IL-11 gene variants occur infrequently, their impact on molecular events during early phases of pregnancy should be further elucidated.

[Comparison of prolactin, free T3 and free T4 levels in the follicular fluid of infertile women and healthy fertile oocyte donors]. / Srovnání hladin prolaktinu, fT3 a fT4 ve folikulární tekutine zen s poruchou plodnosti a zdravých plodných dárkyn oocytu

OBJECTIVE: Follicular fluid (FF) provides an important microenvironment for the development of oocytes. The biochemical composition of the FF plays critical role in the oocyte competence. The aim of our study was to compare the levels of selected hormones in FF of infertile women and healthy fertile oocyte donors. DESIGN: Retrospective comparative study. SETTING: Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague; Department of Gynecology and Obstetrics, Charles University and University Hospital in Pilsen; Institute of Reproductive Medicine and Endocrinology, IVF Centers Prof. Zech, Plzen. METHODS: Levels of prolactin, free T3 and free T4 hormones in the FF of 146 women were analyzed. We have analysed FF of 74 infertile patients (mean age 31 years, SD = 4.65) and 72 healthy fertile oocyte donors (mean age 26 years, SD = 4.44). Only blood free samples were studied after pooling of all FF samples each patient. Levels of hormones were determined using ECLIA method (Electro-Chemi-Luminiscent Immunoassay) on the Cobas e411. RESULTS: RESULTS showed statistically significantly higher levels of prolactin (p=0.0006) and free T4 hormone (p=0.0246) in FF of infertile women in comparison to the group of healthy fertile oocyte donors. CONCLUSION: Our study confirms the presence of prolactin and thyroid hormones in FF and it can be suggested that they play a key role in the regulation of reproductive processes. The study of FF from donors and their detailed comparison with infertile patients with various gynaecological causes of infertility has great value for better understanding of regulatory mechanisms of fertility.

[Comparison of selective oxidative stress parameters in the follicular fluid of infertile women and healthy fertile oocyte donors]. / Srovnání vybraných parametru oxidacního stresu ve folikulární tekutine u zen s poruchou plodnosti a u zdravých fertilních dárkyn oocytu

OBJECTIVE: Follicular fluid (FF) affects oocyte development and disruption of its homeostasis has a crucial effect on egg developmental potential. The aim of this study was to compare the levels of selected oxidative stress markers in the FF of women with impaired fertility and healthy fertile oocytes donors. DESIGN: A retrospective comparative study. SETTING: Faculty of Medicine in Pilsen, Charles University in Prague; Institute of Reproductive Medicine and Endocrinology, IVF Center Prof. Zech, Pilsen. METHODS: Levels of homocysteine (Hcy), malondialdehyde (MDA), glutathione peroxidase (GPx), total antioxidant capacity (AOK) and total protein (CB) were analyzed in the FF. We have analysed FF of 146 women - 74 infertile patients (mean age 31 years, SD = 4.65) and 72 healthy fertile oocyte donors (mean age 26 years, SD = 4.44). Only blood free samples were studied after pooling of all FF samples each patient. RESULTS: The study showed a statistically significantly higher Hcy levels (p < 0.0001) in the FF of healthy fertile women compared with impaired fertility group both - comparing the two groups regardless the age and in groups of the same age range (for the age group between 20 to 29 years isp = 0.0002, for the age group between 30 to 39 years is p < 0.0001). When divided into above age ranges we found statistically significantly higher levels of MDA in the control group aged 20 to 29 years compared to same age infertile patients (p = 0.0374) and statistically significantly higher AOK in infertile women between 30 to 39 years of age compared to same age control group (p = 0.0458). CONCLUSION: The presence or on the contrary the absence of prooxidant parameters in the FF has an important role in the ability of conception and subsequent embryo development.

Comparison of P19-derived neuroprogenitor and naive cell survival after intracerebellar application into B6CBA mice.

Mouse embryonic carcinoma cells (P19 line) were studied for both their survival and developmental potential in the intact cerebellum of B6CBA mice. The P19 cells were cultured and labelled with green fluorescent protein using transfection. Cells were used for transplantation either in the undifferentiated stage or after 3 days of neurodifferentiation induced by retinoic acid. The intracerebellar application was performed in 43 mice: group A (N = 21) received neuroprogenitors and group B (N = 22) received undifferentiated cells. The morphology of transplanted cells within the context of the surrounding cerebellar tissue was evaluated after 3 weeks. Naive P19 cells engrafted and survived in the cerebellum of 7 of the 22 adult mice (survival rate 31.8 %). Neuroprogenitors survived in 13 of the 21 mice (survival rate was 61.9 %). Since the cut-off is P < 0.05, the difference is not statistically significant (P = 0.069). An expansive appearance of the graft was significantly more frequent (P = 0.0047) in naive P19 cells than in neuroprogenitors. In mice in which the grafts did not survive, no marks of grafted cells or only fluorescing detritus were found. In conclusion, this is the first study to track the fate and morphology of embryonic carcinoma cells transplanted into the cerebellum, confirming that neuroprogenitors derived from embryonic carcinoma cells can settle in the host tissue and differentiate according to the surrounding conditions. With further validation, the embryonic carcinoma cells could become a valuable model with which to study the impact of cell therapy on neurodegenerative diseases.

[Contribution of molecular biology to the diagnosis and therapy of colorectal carcinoma--the present and future]. / Prínos molekulární biologie v diagnostice a lécbe kolorektálního karcinomu--soucasnost a budoucnost.

Colorectal cancer is one of the most frequent malignant disease and despite of the development of modern surgical and oncological treatment, it is still a very severe diagnosis for the patient. The survival of the patient after the radical surgery is mostly affected by the time of detection of the disease and by the selection of the appropriate oncological treatment. The effectivity of the oncological treatment depends mainly on the features of the malignant tissue. During the last decade, the importance of the molecular biology and it's methodology have been growing for both detection of the disease and the selection of the best treatment for the individual patient. Genetic and epigenetic characteristics of the tumours helps to predict the prognosis of the disease and also select the best treatment, which extends the disease-free and overall survival of the patient. The presented review describes the most important molecular-biological characteristics with the prognostic or predictive function, which are used in the clinical practice or are in the later phase of clinical study.

Characterization of P19 cells during retinoic acid induced differentiation.

The aim of our study was to characterize mouse embryonal carcinoma (EC) cells P19 in different stages of retinoic acid induced neurodifferentiation by two methods, immunocytochemistry and RT qPCR. The characterization of the cells is crucial before any transplantation into any model, e.g. in our case into the mouse brain with the aim to treat a neurodegenerative disease. Specific protein markers (MAP-2, OCT-4, FORSE-1) were detected by immunocytochemistry in the cell cultures. The mRNA expression levels of PAX-6, MASH-1, Brachyury, GATA-4 and AFP were determined by RT qPCR method. HPRT was used as a housekeeping gene. The degree of differentiation can be characterized by expression of analyzed genes. The presence of OCT-4 and FORSE-1 proteins in undifferentiated pluripotent cells and the presence of dendrite specific MAP-2 in neuroprogenitors was detected. The expression levels of PAX-6 and MASH-1 increased and expression of Brachyury decreased during the neurodifferentiation process. The expression levels of GATA-4 and AFP were the highest after induction of differentiation with retinoic acid. Detailed characterization of cells before transplantation experiments can contribute to better understanding of their effect.

Leukaemia inhibitory factor (LIF) gene mutations in women diagnosed with unexplained infertility and endometriosis have a negative impact on the IVF outcome. A pilot study.

The frequency of functionally relevant mutations of the leukaemia inhibitory factor (LIF) gene in infertile women is significantly enhanced in comparison with fertile controls. The objective of this retrospective cohort study was to evaluate the impact of LIF gene mutations on the outcome of the treatment in women with various causes of infertility. Fifteen infertile women with the G to A transition at position 3400 leading to the valine to methionine exchange at codon 64 were analysed. Group A was made up of women with diagnoses that are frequently accompanied by changes in humoral as well as cell-mediated immunity - idiopathic infertility and endometriosis (N = 7). Group B consisted of patients with polycystic ovary syndrome (PCOS), andrological factor, tubal factor and hyperprolactinaemia (N = 8). The control group comprised 136 infertile women with no LIF gene mutation diagnosed with idiopathic infertility and endometriosis (N = 37) (group C) and patients with PCOS, tubal and andrological factor (N = 99) (group D). Seven of the mutation-positive patients were successfully treated by in vitro fertilization (IVF), but nobody in this group was diagnosed with idiopathic infertility and only one with endometriosis, which means that there is a statistically significant difference in the pregnancy rates between groups A and B (P = 0.01, Fisher's 2 by 2 exact test) but no statistically significant difference when comparing patients with the LIF gene mutation (group A+B) to no LIF gene mutation (group C+D). The results suggest that in mutation-positive women the idiopathic infertility and endometriosis have a negative impact on the outcome of IVF treatment.

[Mediolateral episiotomy and anal sphincter trauma]. / Mediolaterální epiziotomie a poranení análního sfinkteru.

OBJECTIVE: A summary of recent knowledge of the correlation between mediolateral episiotomy and anal sphincter injury. DESIGN: Review. SETTING: Department of Gynaecology and Obstetrics, Charles University and University Hospital Pilsen. CONCLUSIONS: The methodology of most studies is not well managed. Four problematical points were identified: definition of the mediolateral episiotomy, practical execution of the mediolateral episiotomy, diagnostics of perineal trauma and classification of the perineal trauma. Mediolateral episiotomy is often deficiently defined. Definitions differ depending on individual textbooks or departments. The majority of studies gives no definition and no description of the practical execution of an episiotomy or describes it inadequately. To the current knowledge there is no international consensual definition, which is used universally. Until 2003, there was no study evaluating adequate implementation of the mediolateral episiotomy. It appears that most of executed mediolateral episiotomies are not truly mediolateral. The angle of inclination between 40-60 degrees was suggested. According to the latest study, the lower limit of the mediolateral episiotomy definition (40 degrees) appears to be insufficient. At the present time, the correlation between mediolateral episiotomy and perineal trauma cannot be precisely evaluated. Before analyzing the benefits and risks of mediolateral episiotomy, an international consensus must be found, that would establish an exact definition of mediolateral episiotomy.

Beneficial effects of mesenchymal stem cells on adult porcine cardiomyocytes in non-contact co-culture.

Mesenchymal stem cells (MSCs) have been reported to improve survival of cardiomyocytes (CMCs) and overall regeneration of cardiac tissue. Despite promising preclinical results, interactions of MSCs and CMCs, both direct and indirect, remain unclear. In this study, porcine bone marrow MSCs and freshly isolated porcine primary adult CMCs were used for non-contact co-culture experiments. Morphology, viability and functional parameters of CMCs were measured over time and compared between CMCs cultured alone and CMCs co-cultured with MSCs. In non-contact co-culture, MSCs improved survival of CMCs. CMCs co-cultured with MSCs maintained CMCs morphology and viability in significantly higher percentage than CMCs cultured alone. In viable CMCs, mitochondrial respiration was preserved in both CMCs cultured alone and in CMCs co-cultured with MSCs. Comparison of cellular contractility and calcium handling, measured in single CMCs, revealed no significant differences between viable CMCs from co-culture and CMCs cultured alone. In conclusion, non-contact co-culture of porcine MSCs and CMCs improved survival of CMCs with a sufficient preservation of functional and mitochondrial parameters.

Distribution of Connective Tissue in the Male and Female Porcine Liver: Histological Mapping and Recommendations for Sampling.

The pig is a large animal model that is often used in experimental medicine. The aim of this study was to assess, in normal pig livers, sexual dimorphism in the normal fraction of hepatic interlobular and intralobular connective tissue (CT) in six hepatic lobes and in three macroscopical regions of interest (ROIs) with different positions relative to the liver vasculature. Using stereological point grids, the fractions of CT were quantified in histological sections stained with aniline blue and nuclear fast red. Samples (415 tissue blocks) were collected from healthy piglets, representing paracaval, paraportal and peripheral ROIs. There was considerable variability in the CT fraction at all sampling levels. In males the mean fraction of interlobular CT was 4.7 ± 2.4% (mean ± SD) and ranged from 0% to 11.4%. In females the mean fraction of the interlobular CT was 3.6 ± 2.2% and ranged from 0% to 12.3%. The mean fraction of intralobular (perisinusoidal summed with pericentral) CT was <0.2% in both sexes. The interlobular CT represented >99.8% of the total hepatic CT and the fractions were highly correlated (Spearman r = 0.998, P <0.05). The smallest CT fraction was observed in the left medial lobe and in the paracaval ROI and the largest CT fraction was detected in the quadrate lobe and in the peripheral ROI. For planning experiments involving the histological quantification of liver fibrosis and requiring comparison between the liver lobes, these data facilitate the power analysis for sample size needed to detect the expected relative increase or decrease in the fraction of CT.

The combination of nanofibrous and microfibrous materials for enhancement of cell infiltration and in vivo bone tissue formation.

Fibrous scaffolds are desired in tissue engineering applications for their ability to mimic extracellular matrix. In this study we compared fibrous scaffolds prepared from polycaprolactone using three different fabrication methods, electrospinning (ES), electro-blowing and melt-blown combined with ES. Scaffolds differed in morphology, fiber diameters and pore sizes. Mesenchymal stem cell adhesion, proliferation and osteogenic differentiation on scaffolds was evaluated. The most promising scaffold was shown to be melt-blown in combination with ES which combined properties of both technologies. Microfibers enabled good cell infiltration and nanofibers enhanced cell adhesion. This scaffold was used for further testing in critical sized defects in rabbits. New bone tissue formation occurred from the side of the treated defects, compared to a control group where only fat tissue was present. Polycaprolactone fibrous scaffold prepared using a combination of melt-blown and ES technology seems to be promising for bone regeneration. The practical application of results is connected with enormous production capacity and low cost of materials produced by melt-blown technology, compared to other bone scaffold fabrication methods.

Association of the leukemia inhibitory factor gene mutation and the antiphospholipid antibodies in the peripheral blood of infertile women.

To characterize the impact of the potentially functional mutation--the G to A transition at the position 3400 of the leukemia inhibitory factor (LIF; a pluripotent cytokine that plays a central role in the control of the embryo implantation) gene that leads to the exchange of valine with methionine at codon 64 we evaluated the association of the LIF gene mutation and the levels of antiphospolipid antibodies (aPLs) in the peripheral blood of infertile women (the aPLs examination was part of our routine immunological test during the infertility check-up). Eight infertile mutation-positive women were diagnosed with idiopathic infertility (n=5) and endometriosis (n=3) and their levels of aPLs in serum were compared with 115 infertile women without any LIF gene mutation. Enzyme-linked immunosorbent assay was used for the detection of seven antiphospholipid antibodies; the results were statistically assessed by the Fisher's 2 by 2 exact test to evaluate the association of the LIF gene mutations and aPLs in serum of infertile patients. The presence of aPLs was significantly higher in our study group (100%) than in 30% of aPLs-positives in control infertile patients (p = 0.0035) which indicates that the aPLs are elevated in women with LIF gene mutations.

[Evaluation of the development of laparoscopically-assisted vaginal hysterectomy in the Czech Republic over the last 10 years based on the data of the National Registry of Gynecology Laparoscopy Complications]. / Hodnocení vývoje laparoskopicky asistované vaginální hysterektomie v CR za posledních 10 let na základe dat z Národního registru komplikací gynekologické laparoskopie.

OBJECTIVE: To survey the performance of laparoscopically assisted vaginal hysterectomy (LAVH) and other gynecologic laparoscopic operations performed in the Czech Republic. This series covers a period of 10 years, in which 23889 LAVH was performed. DESIGN: Retrospective study (Canadian Task Force classification II-3). METHODS: Two questionnaires were sent each year to more then one hundred of clinics performing laparoscopic gynecologic operations. RESULTS: In 2005, 52% of clinics responded with a total of 4062 LAVH operations. The most frequently reported indication for LAVH in 2005 were uterus myomatosus (58%), uterine bleeding (18%) and adnexectomy (9%). The overall complication rate in 2005 during LAVH was 6.,9% (n = 280), the rate of complications requiring laparotomy was 3.9% (n = 161). Increased experience with LAVH over last 10 years has had two consequences: a significant drop in the number of complications (14.7% in 1996 and 6.9% in 2005) and a drop in the mean time of duration of an operation (101 minutes in 1996 and 82 minutes in 2005). CONCLUSION: LAVH is replacing abdominal hysterectomy, which means significantly less discomfort and shorter hospital stay for patients.