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In spite of its incidence decreasing to 1% nowadays, prosthesis-related infection remains a research, diagnostic, therapeutic and cost-related problem. It can be defined as a presence of bacteria in the artificial joint space, which is significantly associated with evident laboratory and/or tissue markers, and clinical signs of running infection. We believe that the more precise understanding of pathogenesis, the more effective preventative and therapeutic measures, and the lower infection rate. The implants are colonized by airborne, skin-, and/ or surgeon-related bacteria during surgery despite being operated in closely respected operating regime. Some prosthetic characteristics are advantageous and may play important roles in the process of bacterial adherence. After successful attachment on the biomaterial surface bacteria multiply and physiologically transform into a "biofilm" community, making them much more resistant to antibiotic therapy and host immunity. Bacterial resistance is a complex phenomenon influenced by intrinsic and extrinsic factors, including the cell configuration in the biofilm community. So the cure of periprosthetic sepsis without removing of all foreign bodies and necrotic bone fragments is often ineffective. Acute hematogenous sepsis is suggestive of a distortion of a previously aseptic joint space by invasion of bacteria through the vessels.

According to recent research on mice, less on human material, cells responsible for clearing apoptotic cells away during development are, besides non-professional phagocytes, also tissue-fixed macrophages. The aim of our work was the determination of macrophage role in the phagocytosis of apoptotic bodies in neogenous zone of human metanephros. Histologicaly normal kidneys were collected from embryos and fetuses ranging from the 8th-28th week of IUD. These tissues were routinely processed. In the first step we detected CD68+ cells by means of standard indirect three-step immunohistochemical method having used MAb NCL-CD68-KP1 (macrophage marker) to find out whether such cells are actually present. In the second step tissue sections were labelled by double-staining principle (TUNEL technique for the detection of apoptosis and above mentioned macrophage marker) to judge co-localization of these two items. The slides were observed by using immersion objective and the amount of apoptotic cells was expressed in percents. CD68+ macrophages appeared dispersely as single cells or small groups in all the ages studied. According to our results, CD68+ macrophages phagocytose 37-75% of apoptotic cells present in neogenous zone and the number of engulfed apoptotic cells increases in the 12th week of the IUD, i.e. in the early fetal period and later it merely fluctuates.

At present surgery in comparison with other medical fields does not fulfil our ambitions and we are not happy with this state. Surgery is not developing as fast as oncology, haematology, immunology and other branches, which are based on rapidly developing molecular biology. Surgeons are aware of this situation and they do not want to lose. We want to play a part in this new development without losing our position and distinctive surgical goals. To seek these new ways is a goal of the Petřivalský – Rapant Day and we are proud that in our country we are one of the first who want to make this historical change. Therefore in this conference we include topics that should increase our knowledge of malignancies and influence their therapy according to the newest molecular-biological findings.

Nutritionally beneficial compounds naturally present in vegetable lipids will be subject of this minireview. Thisarticle will discuss lipidic compounds from less known vegetable sources and potential advantages of its incorporationinto human diet as a functional ingredient.

Liver injury, and frequent consequent fibrosis, is the focus of a number of research groups ranging from molecular biologists to clinicians. It encompasses many aspects and approaches. Among biochemical events the role of Kupffer cells, oxidative stress and pro-inflammatory mediators are eminent. In this review we focus on recent findings into use of natural substances for the modulation of oxidative burst as well as production of inflammatory mediators by Kupffer cells.

In this article, the author reviews and updates the basis of interactions with warfarin, illustrated with appropriateexamples. The interactions with drugs, medicinal herbs and foods are summarised.

The present article gives a concise survey of contemporary opinions on acne vulgaris, its etiopathogenesis, clinical forms and laboratory diagnostics. In particular, the value of microbiological diagnostics and possibilities of local as well as general therapy are discussed. Moreover, our experience is described with vaccinotherapy to manage serious clinical forms and cases when current therapy fails.

Leptin is a circulating pleiotropic hormone that play an important role in appetite control, fat metabolism, regulationof body weight, fetus growth, growth and aging of adults and hematopoiesis. It is expressed abundantly and specificallyin the adipose tissue. A liver cell with developed steatosis represents a cell metabolism similar to metabolism of cells ofadipose tissue. Analyses of serum leptin and free leptin receptor in the serum of patients with steatosis showed significantvariations from reference limits of normal values. However in liver tissue with verified steatosis detection of mRNAgene for leptin was not proven. Such expression of ob gene for leptin was not found even in the liver tissue without steatosis.With respect to the absence of ob gene expression, the direct effect of ob gene expression on other parameters ofleptin metabolism could not be evaluated. The RT-PCR method with verified specificity and satisfying sensitivity was developed.The results obtained from analysis of serum leptin and free leptin receptor in the serum are presented and evaluated.The used methods were verified and reference limits for Czech population were defined in dependence on age andother clinical parameters.

Interleukin-1 (IL-1) is one of the most potent proinflammatory cytokines involved both in physiologicalimmune responses and in the development of various immunopathological disorders. The members of IL-1 familyform a complex system with tight regulatory mechanisms and clarifying their role in disease remains in the centre ofresearch interests. This review article except of summarising recent findings about IL-1 family members focuses onassociations of IL-1 gene complex polymorphisms with human diseases.

I report and discuss a very rare case of primary involvement of the uterine cervix by hydatid cyst a parasiticdisease caused by Echinococcus granulosus, whose first and most important site is the liver. The case wasmisdiagnosed as an ovarian cyst until the time of operation.And this is to alert the gynaecologist to the possibility of hydatid cyst when a septated mass is found in the pelvis.

The Cathepsin-D proteinase was demonstrated immunohistochemically in cases of intestinal coccidiosis in chickens; itsexpression concerned almost the cyto-invasive stages of reproductive cycle of several strains of coccidia (merozoites of theII-nd generation and also microgametes). The authors consider Cathepsin-D as a potentional „penetration/spreading factor“,being significant for intracellular invasion and spreading of coccidia into host cells. In diagnostic practice, an immunohistochemicaldemonstration of Cathepsin-D is a relatively simple and sensitive method, which brings information onthe extent of parasitic lesions, and on participation of particular evolutive forms of coccidial life cycle; it may also be usefulin proving isolated coccidia in the intestinal epithelium, poorly recognized in routine HE preparation. Some Cathepsin-D protease reactivity in intestinal Cryptosporidiosis is also briefly mentioned. The examined cases of cerebral toxoplasmosisand primary acanthamoebic meningoencephalitis (PAME) showed in above mentioned methods negative results.

Olomouc 29. – 30. 5. 2001

During the long period of cyclosporine-containing dosage forms development a lot of significant findings havebeen done especially in the field of drug delivery systems. Currently available drugs are based, from technologicalpoint of view, on self-dispersible drug delivery systems, which contain cyclosporine solved in pharmaceuticallyacceptable vehicle. One can find difference among particular systems a) at particle size distribution after dispergation,b) at composition and c) at bioavailability of cyclosporine. As far as improvement of bioavailability between originalbrand leader formulation Sandimmune® and recent brand leader formulation Neoral® was followed by significantimprovement in particle size distribution it was generally assumed that the reason for this improvement have beenfound. Several other formulations e.g. Consupren® or SangCyA®– self-dispersible systems, more or less correspondwith above mentioned theory that smaller is better and by this principle bridged liquid based dosage forms withsolid dosage forms. Bioavailability of novel drug delivery system which gives coarse dispersion with average particlesize between 1–150 μm when dispersed have been tested on healthy volunteers. No difference among pharmacokineticparameters of novel drug delivery system and microemulsion system have been observed in spite of fact thataverage particle size of novel system is almost 1000× bigger.

With an increasing number of lipophilic drugs under development, homolipids and heterolipids have gainedrenewed interests as excipients for oral drug delivery systems. Oral administration has many advantages for chronicdrug therapy. It is relatively safe, convenient for the patient and allows self administration.This article is notintended to review the broad area of lipid-based vehicle for oral drug delivery comprehensively. The rationalebehind choosing lipids materials for pharmaceutical dosage forms and their applications is discussed. It alsocomments on the methods for monitoring the physicochemical properties of vehicles and formulations anddescribes a range of pharmacopoeial excipients suitable for these purposes. The excipients selected here arepharmacopoeial in European Pharmacopoeia 4th Ed.,United States Pharmacopoeia 24th Ed./National Formulary19th Ed. and Japanese Pharmacopoeia 13th Ed. or are drafted in Pharmaeuropa and Pharmacopoeial Forum.Widening availability of lipidic excipients with specific characteristics offer flexibility of application with respectto improving the bioavailability of poorly water-soluble drugs and manipulating release profiles.

Olomouc, 7 June 2000

Dedicated to the 60th birthdays of prof. MUDr. Václav Lichnovský, DrSc.The neonatal rat cardiomyocyte model enables heart researchers to study and understand the morphological,biochemical and electrophysiological characteristics of the heart. This model offers a broad spectrum of experiments,such as studies of contraction, ischaemia, hypoxia and the toxicity of various compounds. This reviewexamines the methodology for the isolation and cultivation of primary neonatal rat cardiomyocyte cultures,considers possible problems and pitfalls, and presents an optimized protocol for cardiomyocyte preparation.

Constructs for production of a probe for monitoring of CHS3 expression in saccharomyces cerevisiae

In this study the serum levels of specific anti-nucleosome autoantibodies (autoAb) were measured in the followup of the group of SLE patients and compared to the levels of anti-dsDNA, C3, C4 components of complement, toECLAM- European Lupus Activity Measure and to the levels of sVCAM-1, sICAM-1, neopterin, sIL-2r, IL-10 andfas ligand. Anti-nucleosome antibodies were detected using an enzyme immunoassay for the quantitative determinationof autoAb to nucleosomes and their components in the human serum in the group of 52 patients, 50 of themfemale, who all fulfilled criteria of SLE according to ACR (American Colledge of Rheumatology). Anti-nucleosomeantibodies were followed up during 3 subsequent measurements during six months and compared to the developmentof ECLAM index, the levels of anti ds-DNA antibodies and C3, C4 complement components levels and otheractivity markers.The probands with the stable low disease activity had the lowest levels of anti-nucleosome Ab (83 ±33.0 IU/ml).The patients with the stable high or increasing activity had the mean titers in the first series 200.42 ±36 IU/ml and192 ±65 IU/ml and 188±58.11 IU/ml in the 2. and 3. serial measurement. Subgroup with the high starting activityand with decrease of the activity index had the mean levels of 223 ±39.0IU/ml, 185.6 ±37.6 IU/ml and 207 ±55.5 IU/mlduring three measurements. In the subgroup of patients with lupus nephritis (LN) n=15, the anti-nucleosome Ablevels were 176 ±34.0 IU/ml, 174 ±30.0 IU/ml and 148±32.0 IU/ml. A strong correlation was found between levelsof anti-nucleosome and anti-dsDNA antibodies. All sera with anti-dsDNA antibodies over 40 had at the same timepositive titer of anti-nucleosome antibodies, however, 21 sera with negative anti-dsDNA had positive test for antinucleosomeantibodies. Anti-nucleosome antibodies correlated with ECLAM activity index and reversibly with C4.No correlation was established with levels of C3, sVCAM-1, sICAM-1, IL-10, fas ligand, neopterin and sIL2r. Theexamination of anti-nucleosome Ab contributes to the early diagnosis and is an instrument in the differentialdiagnosis of the connective tissue diseases and it seems to be especially valuable in anti-dsDNA negative lupus. Theuse of anti-nucleosome antibodies measurement as a marker of disease activity deserves further evaluation.

Chemokines mediate their multiple effects by binding to a variety of specific receptors, that comprise a subfamilyof rhodopsin-like, 7-transmembrane domain receptors, coupled to G proteins. Some of these receptors serve ascoreceptors for HIV, some of them could be expressed as markers for T lymphocyte functional differentiation. Thisreview aims at summarising data on chemokine receptors, their function in vivo, their target cells and it alsomentions association of chemokine receptor polymorphisms with human disease.

factorsAn aim has been to find the optimal therapeutic approach for each patient suffering from Hodgkin s lymphoma.The disease with 80% curability rate presents the great success of clinical oncology. There is no doubt that thedevelopment and improvement of therapeutic approaches described in this article has contributed to the excellentresults achieved in the last twenty years. But a lot of problems still remain to be solved by haemato-oncologistsdespite such progress in the treatment. The most important tasks are: 1. to increase the curability of advancedstages, 2. to improve therapeutic results in refractory and relapsing patients, 3. to eliminate or decrease the latetoxicity particularly in the favourable early and intermediate stages.

A total of 85 strains of the genus Acinetobacter were isolated from haemocultures at the Institute of Microbiologyof the Teaching Hospital in Olomouc over the period January 1993 to June 1997. Sixty-two (73.0%) strains of theAcinetobacter calcoaceticus – baumannii complex (Acb complex) were the most frequent. In 3 (3.5%) strains it wasimpossible to decide whether they belonged to the Acb complex. Other acinetobacter species were represented by20 (23.5%) strains. The greatest amount (28.2%) of these strains was collected from the Clinic of InternalMedicine. Leukemias, lymphomas and myelodysplastic syndromes were the most frequent clinical diagnoses20.0%) of the patients with a positive haemoculture. The most effective antimicrobial preparations tested were asfollows: meropenem (98.8% of susceptible strains), colistin (94.1%), quinolones (90.6–94.1% according to the typeof agent) and amikacin (91.8%). The Acb complex strains were less susceptible to antimicrobial agents than otheracinetobacters. Production of inductive chromosomal beta-lactamases AmpC was proved in 42 (49.4%) strainswhilst no occurrence of extended-spectrum beta-lactamases (ESBL) in the isolated organisms was recorded.

The article describes a polymerase chain reaction (PCR) based method for semiquantitative assessment ofmRNA expression for chemotactic cytokines in the lower respiratory tract cells. Cytokine mRNA was detectedusing PCR preceded by reverse transcription of mRNA into cDNA. The procedures of RNA extraction and ofreverse transcription-PCR assay are described in the first part of the article. The second part demonstratesoptimalisation experiments performed to ensure specificity, accuracy and reproducibility of the RT-PCR assay anddiscuss the approach to mRNA quantification using normalisation of cytokine mRNA expression to the expressionof b-actin mRNA.

Renal osteodystrophy is a common finding in patients with renal insufficiency. The maximum of its intensity isfound in hemodialysis patients. Bone densitometry is so far the best method for non-invasive assessment of theextent of the illness. Some densitometric studies in hemodialysis patients have already been published but theirresults differ in prevalence and intensity of renal osteodystrophy. They also demonstrated a slight relationshipbetween intensity of renal osteodystrophy and duration of the dialysis treatment. Opinions vary on the relationshipbetween bone mineral density and markers of bone turnover. This cross-sectional study found high prevalence ofrenal osteodystrophy (Z-score below –1 in 57% of patients) as well as high a number of severely damaged patients(T-score below –2.5 in 40% of patients). It also showed some correlation between bone demineralisation and theduration of dialysis. None from evaluated markers of bone turnover correlated with bone mineral density.

Development of sophisticated noninvasive methods permits the identification of asymptomatic subjets withearly atherosclerosis. High-resolution B-mode ultrasound measurement of the intima-media thickness (IMT) is asafe and reproducible way of quntifying early changes in the arterial wall, mainly in the carotid arteries. Increasedcarotid IMT is associated with major cardiovascular risk factors and seems to be a powerful predictor of cardiovascularevents, especially coronary artery disease.

The budding yeast Saccharomyces cerevisiae has long proved to be a very useful model in cell biology. Its cell morphologyis established and maintained at least in part by the cell wall, a rigid but dynamic structure that affords mechanical protection.Although fungal cell walls represent an unique phenomenon, recent progress in research has shown striking parallelsbetween yeast and mammalian cells in the area of cell morphogenesis and proliferation. Further studies promise to shedcommon light on the processes of cell morphogenesis including the intersections with proliferation control. This reviewfocuses on the recent progress in this promising area in the yeast Saccharomyces cerevisiae.The process of cell wall synthesis in Saccharomyces cerevisiae was reviewed by several authors recently1,2,3. Briefly, thecell wall represents a complex structure of cross-linked chitin, β-(1,6)-d-glucan, β-(1,3)-D-glucan and mannoproteins.Chitin and β-(1,3)-D-glucan are synthesized by enzymatic complexes at the cell membrane and extruded into the periplasmicspace, mannoproteins are synthesized along the yeast secretory pathway, and the site of β-(1,6)-D-glucan synthesis is stillunknown. The principal motif which interconnects individual cell wall constituents was recently identified by Kollár4 etal. The mechanisms of cross-linking of the polymers in the wall remain unknown, however. Recently, nevertheless,substantial progress has been achieved in understanding the signalling pathways which target the cell wall construction.

Some authors described a muscular adult man with a quite large bilateral muscle originating from the front margin of thetransverse process of the C1 – C2 vertebrae and inserting to the back margin of the clavicle next to the conoid tubercle.They named it levator claviculae.

The authors described a unique occurence of the third head of the biceps brachii, pointing out the possibility of attachementof its muscle bundles after partial rupture. This situation could occur, especially when this anomaly was found onlyunilaterally.Dedicated to the 65th anniversary of Doc. MUDr. et RNDr. Milan Černý, CSc.