Standard operating procedures in experimental liver research: thioacetamide model in mice and rats.

In addition to carbon tetrachloride (CCl4), thioacetamide (TAA) represents a second widely used model for the induction of experimental liver fibrosis, but can also be employed for the development of acute liver failure and liver tumours. While TAA itself is not hepatotoxic, its reactive metabolites covalently bind to proteins and lipids thereby causing oxidative stress and centrilobular necrosis. Compared with CCl4, TAA leads to more periportal infiltrates and more pronounced ductal proliferation. While TAA has been shown to induce liver fibrosis development in several different mouse strains, wide variations in the administration routes, doses and treatment durations have been reported. Therefore, an adoption of a universal standard operating procedure for the administration of TAA is urgently needed. For that purpose, we are presenting here two TAA models (intraperitoneal administration of 150 mg/kg of TAA three times per week for 11 weeks in rats, and TAA administration in drinking water at 300 mg/L for 2-4 months in mice) with which we have had success in reliably and reproducibly developing chronic liver injury and fibrosis.

Lipopolysaccharide-induced inflammatory liver injury in mice.

The intraperitoneal application of lipopolysaccharide (LPS) alone or in combination with other hepatotoxins is an experimental model for inducing systemic and hepatic inflammation in rodents applied worldwide. The endotoxin is recognized by the LPS-binding protein. This complex binds together with the lymphocyte antigen 96 (MD2) and the pattern-recognition receptor CD14 to members of the toll-like receptor family. The activated receptor complex in turn transduces signals to well characterized intracellular cascades that result in a multifaceted network of intracellular responses ending in inflammation. The most prominent among these is the activation of the NF-κB pathway and the production of a multitude of inflammatory cytokines. Although the application of LPS is in general easy to perform, unintended variations in preparation of the injection solution or in handling of the animals might affect the reproducibility or the outcome of a specific experiment. Here, we present a well-standardized protocol that allows for an induction of highly reproducible acute hepatic inflammation in mice. Furthermore, examples of appropriate readouts for the resulting inflammatory response are given.

[Long-term outcomes at not-penetrating glaucoma surgery]. / Dlouhodobé výsledky u nepenetrujících operací glaukomu.

OBJECTIVE: To evaluate the long-term compensation and efficacy after non-penetrating deep sclerectomy using different types of implants. METHODS: Patients were divided in 3 groups: deep sclerectomy without implant (DS): 34 eyes, deep sclerectomy with Staar implant (DS+STAAR): 31 eyes, deep sclerectomy with T-flux implant (DS+T): 27 eyes. Postoperatively was evaluated: IOP, therapy and efficacy. RESULTS: The data are described by the relative quantity, continuous data by median, 5th and 95th percentile, always in order of groups: DS; DS+STAAR; DS+T, resp. Age at the time of surgery was 65.6 (44.9; 77.9); 64.5 (44.8; 85.0); 72.1 (51.8; 77.0) years. Follow-up period was 84 (36; 145); 96 (36; 141); 81 (36; 134) months. IOP was 26 (20; 32); 26 (20; 34); 26 (21,34) mmHg postoperatively. IOP was 14 (4; 18); 14 (4; 22); 8 (2; 16) mmHg at 1 month postoperatively. IOP was 17 (13; 23); 17 (12; 21); 14 (12; 18) mmHg at 96 months. IOP was 18 (1; 20); 18 (10; 22); 15 (13; 16) mmHg at 132 months. Completely without therapy were 0; 7.1; 42.9 % of eyes at 120 months. Complete success rate was 0; 7.1; 18.2 % at 96 months. Qualified success rate was 87.5; 89.5; 100 % at 96 months. CONCLUSION: Deep sclerectomy is a good surgical technique to reduce the IOP in patients with glaucoma. Use of T-flux implant provides the best results in our groups.

[Complications of deep nonpenetrating sclerectomy]. / Komplikace u nepenetrujících hlubokých sklerektomií

OBJECTIVE: The purpose of this retrospective study was to evaluate an incidence of complications after deep nonpenetrating sclerectomy with different type of implant. METHODS: A total of consecutive 92 eyes (76 patients) that underwent deep sclerectomy were divided into 3 groups according to the type of nonpenetrating surgery. The first group underwent deep sclerectomy without implant (34 eyes, 29 patients), the second group underwent deep sclerectomy with absorbable collagen implant Staar© (31 eyes, 25 patients) and the third group underwent deep sclerectomy with non-absorbable implant T-Flux© (27 eyes, 22 patients). The complications were divided into 3 types - peroperative, early postoperative and late postoperative. RESULTS: The incidence of complications in above mentioned 3 groups was as follows: Peroperative complications: Perforation of trabeculo-Descemets membrane (2.6; 4.2; 3.7 % respectively). Early postoperative complications: Filtrating operating wound (no occurrence in our groups), Choroideal detachment (17; 19; 16 % resp.), Hypotony (10; 8; 5 % resp.), Hyphaema (7; 3; 0 % resp.), Infectious complications (no incidence of blebitis or endophthalmitis in our groups), Flat anterior chamber (7; 5.7; 5.5 % resp.), Dislocation of implant (1 case of dislocation of Staar©, 1 case of dislocation of T-Flux©). Late postoperative complications: Fibrosis of filtering bleb (30; 36; 24 % resp.), Encapsulated filtering bleb (2.9; 10.3; 0 %), Peripheral anterior synechiae (0; 4.2; 3.7 % resp.), Cataract progression (total incidence 5.5 %), there was no incidence of chronic hypotony, scleral ectasia and late endophthlamitis in our groups. CONCLUSIONS: Deep sclerectomy allows effective lowering of intraocular pressure, without necessity of opening anterior chamber. It brings lower incidence of complications compared to classical penetrating trabeculectomy. Use of implant increases success of surgery due to intrascleral fibrosis reduction. We documented better effect of non-absorbable implant T-Flux© compared to absorbable collagen implant Staar©. The highest incidence of intrascleral fibrosis was in group of patients that underwent deep sclerectomy without implant.

Intermediate filaments take the heat as stress proteins.

Intermediate filament (IF) proteins and heat shock proteins (HSPs) are large multimember families that share several features, including protein abundance, significant upregulation in response to a variety of stresses, cytoprotective functions, and the phenocopying of several human diseases after IF protein or HSP mutation. We are now coming to understand that these common elements point to IFs as important cellular stress proteins with some roles akin to those already well-characterized for HSPs. Unique functional roles for IFs include protection from mechanical stress, whereas HSPs are characteristically involved in protein folding and as chaperones. Shared IF and HSP cytoprotective roles include inhibition of apoptosis, organelle homeostasis, and scaffolding. In this report, we review data that corroborate the view that IFs function as highly specialized cytoskeletal stress proteins that promote cellular organization and homeostasis.

Mallory-Denk-bodies: lessons from keratin-containing hepatic inclusion bodies.

Inclusion bodies are characteristic morphological features of various neuronal, muscular and other human disorders. They share common molecular constituents such as p62, chaperones and proteasome subunits. The proteins within aggregates are misfolded with increased beta-sheet structure, they are heavily phosphorylated, ubiquitinylated and partially degraded. Furthermore, involvement of proteasomal system represents a common feature of virtually all inclusions. Multiple aggregates contain intermediate filament proteins as their major constituents. Among them, Mallory-Denk bodies (MDBs) are the best studied. MDBs represent hepatic inclusions observed in diverse chronic liver diseases such as alcoholic and non-alcoholic steatohepatitis, chronic cholestasis, metabolic disorders and hepatocellular neoplasms. MDBs are induced in mice fed griseofulvin or 3,5-diethoxycarbonyl-1,4-dihydrocollidine and resolve after discontinuation of toxin administration. The availability of a drug-induced model makes MDBs a unique tool for studying inclusion formation. Our review summarizes the recent advances gained from this model and shows how they relate to observations in other aggregates. The MDB formation-underlying mechanisms include protein misfolding, chaperone alterations, disproportional protein expression with keratin 8>keratin 18 levels and subsequent keratin 8 crosslinking via transglutaminase. p62 presence is crucial for MDB formation. Proteasome inhibitors precipitate MDB formation, whereas stimulation of autophagy with rapamycin attenuates their formation.

Intermediate filament cytoskeleton of the liver in health and disease.

Intermediate filaments (IFs) represent the largest cytoskeletal gene family comprising approximately 70 genes expressed in tissue specific manner. In addition to scaffolding function, they form complex signaling platforms and interact with various kinases, adaptor, and apoptotic proteins. IFs are established cytoprotectants and IF variants are associated with >30 human diseases. Furthermore, IF-containing inclusion bodies are characteristic features of several neurodegenerative, muscular, and other disorders. Acidic (type I) and basic keratins (type II) build obligatory type I and type II heteropolymers and are expressed in epithelial cells. Adult hepatocytes contain K8 and K18 as their only cytoplasmic IF pair, whereas cholangiocytes express K7 and K19 in addition. K8/K18-deficient animals exhibit a marked susceptibility to various toxic agents and Fas-induced apoptosis. In humans, K8/K18 variants predispose to development of end-stage liver disease and acute liver failure (ALF). K8/K18 variants also associate with development of liver fibrosis in patients with chronic hepatitis C. Mallory-Denk bodies (MDBs) are protein aggregates consisting of ubiquitinated K8/K18, chaperones and sequestosome1/p62 (p62) as their major constituents. MDBs are found in various liver diseases including alcoholic and non-alcoholic steatohepatitis and can be formed in mice by feeding hepatotoxic substances griseofulvin and 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). MDBs also arise in cell culture after transfection with K8/K18, ubiquitin, and p62. Major factors that determine MDB formation in vivo are the type of stress (with oxidative stress as a major player), the extent of stress-induced protein misfolding and resulting chaperone, proteasome and autophagy overload, keratin 8 excess, transglutaminase activation with transamidation of keratin 8 and p62 upregulation.

Monitoring of epithelial cell caspase activation via detection of durable keratin fragment formation.

Keratins 18 and 19 (K18/K19) are epithelial-specific intermediate filament proteins. Apoptosis induces caspase cleavage at the highly conserved K18 or K19 Asp237, which in K18 is preceded by cleavage at Asp396. We characterized the keratin N-terminal fragments that are generated upon caspase digestion of K18/K19 at Asp237 in order to study keratin dynamics during apoptosis. This was carried out by generating and characterizing antibodies selective to K18/K19 Asp237. K18 or K19 peptides that expose Asp237 in 234VEVD were used for rabbit immunization. The generated antibodies recognized cleaved but not intact K18/K19, exclusively, as determined by blotting or immunofluorescence staining of apoptotic human HT29 cells or livers isolated from Fas-Ab-injected mice. Antibodies to K18/K19 Asp237 recognized the common VEVD-motif as determined by immunoblotting of cells transfected with K18, K19 or K20. The K18/K19 VEVD-directed antibodies demonstrated sequential Asp396 then Asp237 K18 cleavage during apoptosis. Specific-keratin selectivity of the anti-Asp237 antibodies was confirmed by their inability to recognize K14 after UV-induced apoptosis in transfected cells. The Asp237-containing apoptotic keratin fragments are secreted into the medium of cultured HT29 cells and are stable up to 96 h after inducing apoptosis. Furthermore, the generated antibodies recognize keratin apoptotic fragments in sera of mice undergoing hepatocyte apoptosis and sera of patients with cirrhosis, and also recognize apoptotic cells in various epithelial human tumours. Therefore, the N-terminal caspase-generated K18 fragment is stable in tissues and biological fluids. The Asp237-directed antibodies provide a powerful tool to study apoptosis in human and mouse tissues, cells and serum, using a broad range of detection modalities.

Autophagy modulates keratin-containing inclusion formation and apoptosis in cell culture in a context-dependent fashion.

The major pathways for protein degradation are the proteasomal and lysosomal systems. Derangement of protein degradation causes the formation of intracellular inclusions, and apoptosis and is associated with several diseases. We utilized hepatocyte-derived cell lines to examine the consequences of the cytoplasmic hepatocyte Mallory-Denk body-like inclusions on organelle organization, autophagy and apoptosis, and tested the hypothesis that autophagy affects inclusion turnover. Proteasome inhibitors (PIs) generate keratin-containing Mallory-Denk body-like inclusions in cultured cells and cause reorganization of mitochondria and other organelles, autophagy and apoptosis. In cultured hepatoma cells, caspase inhibition blocks PI-induced apoptosis but not inclusion formation or autophagy activation. Autophagy induction by rapamycin decreases the extent of PI-induced inclusions and apoptosis in Huh7 and OUMS29 cells. Surprisingly, blocking of autophagy sequestration by 3 methyl adenine or beclin 1 siRNA, but not bafilomycin A1 inhibition of autophagic degradation, also inhibits inclusion formation in the tested cells. Therefore, autophagy can be upstream of apoptosis and may promote or alleviate inclusion formation in cell culture in a context-dependent manner via putative autophagy-associated molecular triggers. Manipulation of autophagy may offer a strategy to address the importance of inclusion formation and its significance in inclusion-associated diseases.

Preoperative transforming growth factor-beta 1 (TGF-beta 1) plasma levels in operable breast cancer patients.

OBJECTIVES: The aim of this project was to search for new risk prognostic markers in the early stage of breast cancer. We tested preoperative plasma transforming growth factor - beta 1 (TGF- beta 1) levels in patients with operable breast cancer. Correlation with traditional prognostic markers and with positivity/negativity sentinel lymph node was evaluated. MATERIALS AND METHODS: Between 2003 and 2005, 36 patients with operable breast cancer (T1-2, N0-1, M0) with positive or negative sentinel lymph nodes were evaluated for their plasma TGF-beta 1. Twenty-seven healthy individuals (9 premenopausal and 18 postmenopausal) served as controls. Patients were evaluated for the traditional prognostic markers including tumor characteristics, positivity and negativity of sentinel lymph node, TNM, tumor grade, expression of tumor markers CA 15-3 and CEA, hormonal status (pre- or postmenopausal patients, estrogen and progesteron receptor expression), ERB and p53 expression. Predictive value of TGF-beta 1 level and correlation with either of the assessed parameters was tested by one way ANOVA analysis. RESULTS: Measurements of preoperative plasma TGF-beta 1 levels in patients with operable breast cancer were significantly higher compared with healthy individuals (median 15293 and 3983 pg/ml p < 0.0001). TGF-beta 1 level in plasma of patients with a positive sentinel lymph node was significantly higher than in patients with negative sentinel lymph nodes (high vs low, median 18,9 and 14,5 ng/ml, respectively, p = 0.05). CONCLUSION: The determination of TGF-beta 1 status might help to identify a high-risk population early in tumor progression, for which a more appropriate therapy should be established. In the node-negative population, the up-regulation of TGF-beta 1 might constitute an early event that promotes further progression of breast tumors.

[The clinical significance of sentinel lymph node micrometastases in breast cancer]. / Klinický význam mikrometastáz karcinomu prsu v sentinelové uzline.

OBJECTIVE: The advent of sentinel lymph node biopsy and improvements in histopathological and immunohistochemical analysis has increased the rate at which micrometastases are identified. However their significance has been the subject of much debate. Published studies have reported divergent results regarding the significance and implications of axillary lymph node micrometastases. Some studies demonstrate no associations, whilst others have found these to be indicators of poor prognosis, associated with non-SLN involvement, in addition to local and distant failure. The objective of our study was to evaluate the impact of sentinel lymph node micrometastatic cancer to prognosis of the disease. DESIGN: Retrospective study. SETTING: Departments of Gynecology and Obstetrics, Faculty Hospital, Prague. METHODS: From January 2000 to December 2006 in 87 cases with sentinel axillary node negative cancer we reexamined the axillary tissue blocks by serial sectioning, haematoxilin-eosin staining and immunohistochemistry. Additional 15 cases of micrometastatic sentinel node involvement detected by frozen section were included. The overall and disease free survivals of patients with sentinel negative status (N0-67 cases) and with sentinel node micrometastases (Nmic-35 cases) were evaluated. The median follow-up was 60 months (24-96 months). RESULTS: Micrometastases (Nmic) were found in 20 cases (23%). From the group of 67 nodes negative patients (N0) in 7 cases (10.5%) developed tumor recurrence and from the group of 35 Nmic in 5 cases developed five tumor recurrences (13.3%). In the group of N0 patients developed 2 regional recurrences and 3 patients died, but 2 patients died of other causes. In the group of Nmic developed one regional recurrence and 3 patients died of generalization of disease. CONCLUSION: Our study demonstrated that the presence of sentinel node micrometastases is associated with risk of development of distant metastases and generalization of the disease, but not with higher risk of regional recurrence.

[Detection of postoperative lymphoedema in patients with breast cancer]. / Detekce pooperacního lymfedému u pacientek s karcinomem prsu.

OBJECTIVE: Lymphoedema is a severe postoperative complication after treatment of many malignancies. It is a pathological accumulation of extracellular water (ECW). Early diagnostic tool is needed. Multifrequency bioimpedance analysis (MFBIA) is a method for detection of changes in ECW. TYPE OF STUDY: Prospective study. SETTING: Dept. of Obstetrics and Gynaecology of the 2nd Medical Faculty, Charles University, Prague. METHODS: We measured a control group of 72 women and a group of 74 patients undergoing a breast cancer surgery during 18 month after the surgery by MFBIA and circumferency. Characteristics of the patients were recorded. The detection of lymphoedema was done using MFBIA, circumferency measurement and upon the symptoms of the patients. RESULTS: The average age in the controll and tested group was 40,1 and 58,9 years. The average size of the tumour, grade and positivity if oestrogen receptors was 15,1 mm, 2,04 and 43%. In 23 patients (21%) complete lymphadenectomy was performed, in 51 patients (79%) a detection of sentinel lymph node was performed. Lymphoedema was detected in 8 women (11%). In these patients MFBIA detected lymphoedema 9 month earlier in total than other methods. CONCLUSION: MFBIA is a low-cost and precise method for the detection od early stage postoperative lymphoedema. We recommend to incorporate MFBIA into standard dispensatory plan of every patient combined with circumferency measurement.

[A set of patients with breast cancer associated with pregnancy]. / Soubor pacientek s karcinomem prsu v souvislosti s tehotenstvím.

OBJECTIVE: The aim of our study is to follow patients diagnosed with breast cancer associated with pregnancy. DESIGN: Prospective follow-up study. SETTING: Department of Obstetrics and Gynecology of the 2nd Medical Faculty of the Charles University and Faculty Hospital Motol, Prague. METHODS: Between the year 2002 and 2006 12 women were reffered to our department, seven with the diagnosis of breast cancer during pregnancy, three within one year after the delivery, two pregnant after the therapy for breast cancer. A managment was sellected individually with respect to the age of pregnancy, the stage of the disease and to the decision of the women. The average age of the diagnosis, managment, way of delivery, fetal and maternal outcome were recorded. RESULTS: The average age was 31.7 years. Two patients underwent a termination of the pregnancy, other delivered a healthy newborn. Two of our patients are dead, three have a metastatic disease, one was lost for follow-up, the rest are further followed. CONCLUSIONS: Breast cancer diagnosed during pregnancy is a special and rare situation, which needs to be managed individually as there is a lack of information about such patients. These patients should be concentrated in specialized centers.

[Transforming growth factor-beta 1 as a marker in patients with operable breast cancer]. / Transformacní rustový faktor--beta 1 jako marker u pacientek s operabilním karcinomem prsu.

UNLABELLED: OBJECTIVE AND DESIGN OF STUDY: Determination of TGF-beta 1 levels in plasma of patients with operable breast cancer. The correlation of TGF levels with the stage of disease and other prognostic markers. Prospective study. METHODS: There were 36 patients fulfilling the entrance criteria included in the study. The blood samples were taken to set the plasma levels of TGF-beta before the operation, with no adjuvant therapy. 27 age matched healthy volunteers served as controls. The study was approved by ethical board and patients signed informed consent regarding blood sampling and results presentation. Differences between groups were determined using the Mann-Whitney U-test. RESULTS: We proved that TGF-beta 1 levels are elevated in patients with operable breast cancer. Moreover, TGF-beta in plasma of patients with positive sentinel lymph node was significantly higher as compared to patients with negative sentinel node. Most important is the fact that patients involved in our study were in very early stages of disease. CONCLUSION: TGF-beta 1 is marker correlating with breast cancer disease risk factors, especially with positive sentinel lymph node. We conclude that TGF is one of the early markers which may help define the risk of disease progression already before the operation.

Less radical fertility-sparing surgery than radical trachelectomy in early cervical cancer.

The purpose of this pilot study was to determine feasibility and safety of a novel and less radical fertility-preserving surgery; laparoscopic lymphadenectomy with sentinel lymph node identification (SLNI) followed by large cone or simple trachelectomy. Obstetrical and oncologic outcomes were evaluated. Twenty-six patients (6-IA2, 20-IB1) selected on basis of favorable cervical tumor characteristics and the desire to maintain fertility underwent laparoscopic SLNI, frozen section (FS), and a complete pelvic lymphadenectomy as first step of treatment. All of nodes were submitted for microscopic evaluation (sentinel nodes for ultramicrostaging). After a 7-day interval, large cone or simple vaginal trachelectomy was performed in patients with negative nodes. The average of sentinel nodes per side was 1.50 and the average of total nodes was 28.0. Four FS were positive (15.4%). In these cases, Wertheim radical hysterectomy type III was immediately performed. We had no false-negative SLN neither on FS nor on final pathology assessment. Median follow-up was 49 months (18-84). One central recurrence (isthmic part of uterus) was observed 14 months after surgery. This patient was treated with radical chemoradiotherapy, and there was no evidence of the disease 36 months after treatment. Fifteen women planned pregnancy, 11 women became pregnant (15 pregnancies), and 7 women delivered eight children (one in 24 weeks, one in 34 weeks, one in 36 weeks, and five between 37 and 39 weeks). We conclude that lymphatic mapping and SLNI improves safety in this fertility sparing surgery. Large cone or simple trachelectomy combined with laparoscopic pelvic lymphadenectomy can be a feasible method with a high successful pregnancy rate.

[Sentinel lymph nodes identification in vulvar cancer--methods and technique]. / Detekce sentinelových uzlin u karcinomu vulvy--metody a technika.

OBJECTIVE: To evaluate detection of sentinel lymph nodes (SLN) in squamous vulvar cancer with blue dye and 99mTc. The study describes technique of application, timing, management, detection rate (DR), specific side detection rate (SSDR) and false negative rate. DESIGN: Prospective clinical study. SETTING: Department of Obstetrics and Gynecology, Charles University Prague, 2nd Medical Faculty, Teaching Hospital Motol. PATIENTS AND METHODS: 46 women with squamous cell carcinoma tumors stage I or II, <4 cm with no clinical suspect lymph nodes were included. Blue dye alone was used in 16 women and the combination of 99mTc and blue dye was used in 30 women. Radiocoloid 99mTc was applied 3-5 hours and blue dye 3-5 minutes prior to ingvinal incision. RESULTS: We detected 88 SLN in 61 inguinal spaces. The detection rate in the blue dye group was 68.8 % (11 cases). One false negative SLN (6.3 %) appeared in this group. In blue dye+ 99mTc group detection rate was 100 % with no false negative SLN. CONCLUSION: Detection of SLN in squamous vulvar cancer with the combination of 99mTc and blue dye was statistically significantly more effective than using the blue dye alone.

[Fertility sparing surgery in early cervical cancer today and tomorrow]. / Fertilitu zachovávající operace u casného stadia karcinomu delozního hrdla dnes a zítra.

OBJECTIVE: Describtion and evaluation of proposed protocol for conservative, fertility sparing surgeries in the treatment of early stages of cervical cancer and its comparison to most frequently used protocols regarding its reproductive and oncological outcomes. DESIGN: Prospective clinical study. SETTING: Department of Obstetrics and Gynecology, Charles University Prague, 2nd Medical Faculty, University Hospital Motol. PATIENTS AND METHODS: 24 patients that fitted into the inclusion criteria were recruited in the study (6 in stage T1A2 and 18 T1B1). Inclusion criteria were: tumor less than 20 mm in largest diameter, less than 1/2 of the stromal invasion, SCC serum levels within normal range, signed informed consent. Minimum follow up was 24 months. Described therapeutic protocol, evaluation of oncological and reproductive outcomes and comparison with different fertility sparing regimens were performed. RESULTS: In four cases (16.7%) peroperative histopatghological evaluation (frozen section: FS) detected metastatic involvement of lymphatic nodes. In one case (4.1%) following 14 months since initial surgery, reccurent disease in uterine isthmus was detected. The patient underwent chmotherapy and she is 36 months in complete remission. In the series of 15 women planning pregnancy 11 subjects become pregnant so far. Eight deliveries occurred (one in 24th week of gestation--650g, one in 34w--2240g, one in 36w--2700g and five between 37-39w). Only one newborn was delivered in the cathegory of extreme prematurity. CONCLUSION: Sentinel node detection involving peroperative histopathological evaluation followed by subsequent serial processing and together with 2-step therapeutic management increases efectivity of fertility sparing surgeries. Less radical surgery on uterine cervix: reconisaton (in stage IA2) or simple trachelectomy (in early stage T1B1) with negative finding after laparoscopic lymph node dissection has comparative results with radical trachelectomy and abdominal radical trachelectomy. Pregnancy rates and pregnancy outcomes particularly regarding in term delivery is the best after treatment of early stages of cervical cancer using proposed protocol, while very promising results are published in studies with radical vaginal trachelectomy as well. Results published for the patients treated with abdominal radical trachelectomy are unsatisfactory.

Radioguided occult lesion localisation in combination with detection of the sentinel lymph node in non-palpable breast cancer tumours.

PURPOSE OF THE INVESTIGATION: The aim of study was to determine the efficacy of radioguided occult lesion localisation (ROLL) for non-palpable invasive breast cancer combined with sentinel lymph node biopsy (SLNB) and to compare the amount of tissue excised by radioguided navigation versus the hook-wire technique. METHODS: We injected 45 MBq of radiolabelled technetium intratumourally and 15 MBq subdermally 18 hours before surgery in 21 women with bioptically verified non-palpable breast cancer. We identified by gamma probe non-palpable tumours, which were excised, followed by identification and excision of the sentinel lymph node. We compared our results with a group of 12 women with non-palpable lesions marked by hook-wire localisation. RESULTS: ROLL combined with SLNB was successful in 100%; volume of excised tissue was smaller in the hook-wire group but expressed higher variability in volume than in the ROLL group although the difference was not statistically significant. CONCLUSION: The method of ROLL combined with SLNB is technically possible and safe, resulting in minimisation of the surgical intervention and a decrease in postoperative morbidity. ROLL was more precise than the hook-wire procedure even though the amount of tissue excised was the same in both groups.