[Contemporary technicqes in diagnostic patients with vaginal microbiota]. / Soucasné vyuzití technik v diagnostice recidivujícího vulvovaginálního dyskomfortu.

OBJECTIVE: Review of literature dealing with the specificity of the vagina from the standpoint of microbial colonization. DESIGN: Literature review. SETTING: Department of Obstetrics and Gynecology, Department of Clinical Microbiology, University Hospital and Medical Faculty in Hradec Kralove, Charles University Prague. METHODS: Analysis of the literature review and personal experience dealing with vaginal microbiota.

[Hypersensitivity reactions to carboplatin and paclitaxel - our five-years experiences]. / Hypersenzitivní reakce po podání karboplatiny a paklitaxelu - nase petileté zkusenosti.

OBJECTIVE: To analyze hypersensitivity reactions to carboplatin and paclitaxel in patients treated with systemic administration of chemotherapy (carboplatin and/or paclitaxel). DESIGN: Retrospective study. SETTING: Department of Obstetrics and Gynecology, Charles University in Prague, Faculty of Medicine and University Hospital Hradec Kralove. METHODS: One hundred-forty patients treated with systemic administration of chemotherapy were enrolled to our study between years 2008 and 2012. The presence and the grade [grade (G) 1-5; 1 = moderate, 5 = death] of hypersensitivity reactions (HSRs) were evaluated, as well as the influence of some clinical parameters on development of HSR. RESULTS: In total 29 HSRs in 21 patients were analyzed. To carboplatin were reported 19 (66%) HSRs: 13 (45%) HSRs of G1-G3 and 6 (21%) HSRs of G4. To paclitaxel were reported 10 (34%) HSRs: 9 (31%) HSRs of G1-G3 and 1 (3%) HSR of G4. The number of administered cycles of carboplatin to develop G1-G4 resp. G1-G3 HSR was higher in comparison with number of cycles to develop HSR of the same grade to paclitaxel(p = 0.001, resp. p = 0.01). CONCLUSION: HSR to carboplatin is unlike paclitaxel affected by the number of administered cycles. This fact should be included in the clinical management of patients treated with intravenous chemotherapy using carboplatin.

[What is the current stage of vulvovaginal discomfort diagnostics in the Czech Republic? Pilot analysis]. / Jaký je aktuální stav diagnostiky vulvovaginálního dyskomfortuv Ceské republice? Pilotní analýza.

OBJECTIVE: To find out the level of the diagnostic effort of gynaecologists which is focused on the issue of vulvovaginal discomfort. DESIGN: Pilot questionnaire study. SETTINGS: Department of Obstetrics and Gynecology, Department of Clinical Microbiology, University Hospital and Medical Faculty Hradec Králové, Charles University, Prague, Department of Biological and Medici Sciences, FaF UK in Hradec Králové. METHODS AND RESULTS: The evaluation of selected parametres of entrance questionnaire in patients with chronic vulvovaginal discomfort (itching, burning, discharge, vulvodynia more than 4 times a year). It has been confirmed, with the questionnaires, that almost no gynaecologists are interested in the issue of this matter. CONCLUSION: After evaluating the questionnaires a diversion of patient-tailored attitude has been found out, which can lead to negative consequences, particularly, in the future. Overuse of antibacterial and antimycotic medication and blind treatment have been prevailing.

Antibacterial activity of N-benzylsalicylthioamides.

The in-vitro biological activity of N-benzylsalicylthioamides against 8 bacterial strains was determined by broth microdilution method; results were compared with those obtained with neomycin, penicillin G, ciprofloxacin and penicillin V. The compounds showed moderate to high activity against G(+) bacteria; especially compounds 4, 6, 13, 16-21 and 24 exhibited comparable or higher activity than reference drugs. The antibacterial activity was analyzed by quantitative structure-activity relationship (QSAR). The antibacterial activity increased with lipophilicity, with the presence of halogens and with increasing value of Hammet substituent constant σ.

[Vulvodynia and the possibility of its management]. / Nový pohled na problematiku vulvodynie.

OBJECTIVE: A study of vulvodynia and the possibility of its management. SUBJECT: Original study. SETTING: Department of Obstetrics and Gynecology, Teaching Hospital and Medical Faculty Hradec Králové, Charles University, Prague. Department of Neurology, Teaching Hospital and Medical Faculty Hradec Králové, Charles University, Prague. Outpatient psychiatric department Galenus, Hradec Králové. Department of Clinical Microbiology, Teaching Hospital and Medical Faculty Hradec Králové, Charles University, Prague. Department of Biological and Medical Sciences, Faculty of Pharmacy Hradec Králové, Charles University, Prague. METHODS: Analysis and discussion focused on our experience in a long time follow-up of patients with chronic vulvovaginal discomfort and vulvodynia. CONCLUSION: Especially the management of indolent form of vulvodynia would be taken into consideration of idiopathic origin and problematic management. Our study emphasizes the necessity of individual approach. The goal of therapy in this most problematic form is the improving of the actual quality of life. In the case of idiopathic forms of vulvodynia.

[Prognostic factors of mesenchymal and mixed tumors of uterus]. / Prognostické faktory u mezenchymálních a smísených nádoru delozního tela.

OBJECTIVE: Analysis of clinical and histopathological prognostic factors with emphasis on carcinosarcomas, leiomyosarcomas and endometrial sarcomas of uterus. SUBJECT: Review. SETTING: Department of Obstetrics and Gynaecology, Medical Faculty Hradec Králové, Charles University, Prague; The Fingerland Department of Pathology, Medical Faculty Hradec Králové, Charles University, Prague; Department of Clinical Oncology, Medical Faculty Hradec Králové, Charles University, Prague; Department of Biological and Medical Sciences, Faculty of Pharmacy, Hradec Králové; Charles University, Prague. SUBJECT AND METHOD: Discussion about our experience and current evidence. CONCLUSION: Mesenchymal and mixed tumors of uterus are characterized by several prognostic factors as well. Except clinical prognostic variables, histopathological and molecular biological factors may play a role. The most important prognostic significance carry stage of disease, age and performance status of patient and sufficient therapy, especially surgery. In particular tumors some other prognostic factors are recognized. Except cases in really early stage of disease there is usually short interval between end of primary therapy and tumor recurrence. With the exception of endometrial stromal sarcoma, prognosis of these tumors remains very poor.

[Recurrent vulvovaginal candidiasis--possibility of its treatment]. / Rekurentní vulvovaginální kandidóza--soucasný stav problematiky a lécebné moznosti.

OBJECTIVE: A review of recurrent vulvovaginal candidiasis and the possibility of its treatment. SUBJECT: Original study. SETTING: Department of Obstetrics and Gynaecology, Medical Faculty Hradec Králové, Charles University, Prague. Department of Clinical Microbiology, Medical Faculty Hradec Králové, Charles University, Prague. Department of Biological and Medical Sciences, Faculty of Pharmacy Hradec Králové, Charles University, Prague. Department of Clinical Imunology and Allergology, Medical Faculty Hradec Králové, Charles University, Prague. Department of Psychiatry, Medical Faculty Olomouc, Palacky University, Olomouc. METHODS: Analysis and discussion focused especially on our results and experience in a long time followup of patients with confirmed recurrent vulvovaginal candidiasis. CONCLUSION: Owing to the multifactorial character of etiopathogenesis, the management of recurrent vulvovaginal candidiasis would be taken into consideration the complexity of the disease, not only the treatment of individual episodes with antimycotics.

[Multidisciplinary approach to the management of vulvovaginal discomfort]. / Problematika vulvovaginálního dyskomfortu z hlediska mezioborové spolupráce.

Vulvovaginal discomfort is generally the most common reason for visiting a gynaecological clinic. The acute complaints are, in most cases, solved by a gynaecologist himself and the decision procedure is based on possibilities arising from outpatient examination. If need be, the evaluation of wet preparation should be carried out. In many cases we have been confronted with discrepant diagnosis where severe difficulties are not accompanied by relevant development of inflammatory changes and thus the diagnosis gives us an impression of quandary. In this case microbiological examination is the logic solution. With regard to changing living conditions we can expect in the future, that clinical symptoms of vulvovaginitis will not be entirely clear. The overview article points out the possibilities, which a gynecologist is able to use within differential diagnosis in the outpatient department. It also emphasis the necessity of interdisciplinary co-operation, particularly in patients with chronic difficulties where erudite evaluation of fixed preparation and culture examination are beneficial.

[Antifungal effect in selected natural compounds and probiotics and their possible use in prophylaxis of vulvovaginitis]. / Antifungální ucinek u vybraných prírodních látek a probiotik, jejich mozností pouzití v profylaxi vulvovaginitidy.

OBJECTIVE: a survey of effective natural compounds and probiotics with antifungal activity and their possible use in vulvovaginitis as a prophylactic or therapeutic drug. DESIGN: review of literature. SETTING: Charles University at Prague, Pharmaceutical Faculty at Hradec Kralove. METHODS: and review of literature and publications from medical data-bases. CONCLUSIONS: certain natural compounds were proved to exert antimycotic effects. The efficiency of garlic extract under in vitro conditions against yeast of C. albicans family becomes manifest within one hour. Essential oils present in the Cayenne pepper (Capsodum frutescens) exert considerable antifungal activity against C. albicans as well. Clove exerts lower antimycotic effects. The effects of Chinese leek are similar to garlic. Propolis also possesses antifungal properties, and sensitivity of the yeast strains decreases in the following order: C. albicans > C. tropicalis > C. crusei > C. guilliermondii. Its efficiency depends on the composition and place of origin. Tea trea oil (Melalauca alternifolia) is inhibiting strains of Candida family (genus) in minimal inhibiting concentration, higher concentrations are necessary for C. crusei and C. norvengensis. In the prophylaxis of vaginal candidoses, oral administration of the strains of Lactobacilus rhamnosus GR-1 and L. fermentum RC-14, B-54 may be recommended. Moreover there are suitable strains of L. brevis CD-2, L. salivarius FV-2, L. gasseri MB 335, L. plantarum, useful in the form of gel capsules (L. plantarum) or vaginal tablets (L. gasseri). The strains of L. acidophilus CRL 1294 and L. salivarius CRL 1328 display the capacity of self aggregation and co-aggregation with Candida. It should be pointed out that the probiotic strain of Lactobacillus (especially L. casei and L. rhamnosus) may, in exceptional cases, cause bacteriemia and sepsis, especially in immunodeficient patients and those who underwent surgery of alimentary tract. No protective effect against yeast infection was detected with vitamin E, any positive effect was not unequivocally confirmed with zinc, but adjusting zinc levels to normal may be recommended.

[Systemic immunity in patients with recurrent vulvovaginal candidiasis]. / Systémová imunita u pacientek s rekurentní vulvovaginální kandidózou.

OBJECTIVE: This study was undertaken to characterize systemic immunity in the patients with recurrent vulvovaginal candidiasis. SUBJECT: Original prospective study. SETTING: 1. Department of Biological and Medical sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Kralove, 2. Department of Obstetrics and Gynaecology, Medical Faculty, Hradec Kralove, Charles University in Prague, 3. Department of Clinical Immunology and Allergology, University Hospital, Hradec Kralove, 4. Department of Clinical Microbiology, University Hospital, Hradec Kralove, Czech republic. SUBJECT AND METHOD: Basic immunological parameters in 44 patients with recurrent vulvovaginal candidiasis were investigated and analyzed in this longitudinal follow-up study. CONCLUSION: Although there is no fundamental disturbance in systemic immunity in patients with recurrent vulvovaginal candidiasis and the majority of systemic immunological parameters in the patients were within normal range, some subtle changes of a few parameters of humoral and cellular immunity in contrast to the controls were identified.

[Antifungal immunity and its mechanism in female genitalia]. / Protihoubová imunita a její mechanismy ve vztahu k zenskému genitálu.

OBJECTIVE: A review of immunological aspects of fungal infections with focus on vulvovaginal candidiasis. DESIGN: Review. SETTING: Department of Obstetrics and Gynaecology, Medical Faculty, Hradec Kralove, Charles University in Prague, Department of Biological and Medical sciences, Faculty of Pharmacy, Hradec Kralove, Charles University in Prague, Department of Clinical Immunology and Allergology, University Hospital, Hradec Kralove, Department of Clinical Microbiology, University Hospital, Hradec Kralove. SUBJECT AND METHOD: Survey and discussion of current immunological aspects of vulvovaginal candidias based on literature review and personal experience. CONCLUSION: The current knowledge of immunity to Candida vulvovaginitis, particularly from experimental studies on animal models, support a more relevant role of local protection mechanisms than systemic immunity. As a consequence of this compartmentalization of the immune response, the role of innate and adaptive immunity remains unclear. The development of vulvovaginal candidiasis depends on a number of factors, including both the individual predisposition of a macroorganism and the properties of a fungal strain, and the environmental conditions influencing a delicate balance between the host and the yeast.

[Histopathologic prognostic factors, serum markers and selected molecular biologic factors in ovarian carcinoma]. / Histopatologické prognostické faktory, sérové markery a vybrané molekulárne biologické faktory u ovariálního karcinomu.

OBJECTIVE: A review of histopathological prognostic factors, serum markers and selected molecular biological factors in ovarian cancer. DESIGN: Review. SETTING: Department of Obstetrics and Gynaecology, Medical Faculty Hradec Králové, Charles University, Prague; Department of Biological and Medical sciences, Pharmaceutical Faculty, Hradec Králové, Charles University, Prague; Department of Clinical Oncology, Medical Faculty Hradec Králové, Charles University, Prague; Bristol-Myers Squibb, Department of Oncology, Prague. METHOD: Discussion about current evidence from literature. CONCLUSION: Except clinical prognostic factors the wide spectrum behaviour of epithelial ovarian cancers is reflected by some histopathological prognostic factors, serum markers and some molecular biological factors. The most important are grade and tumor ploidy, the level of CA 125 and its changes during the treatment, mutations of p53, BRCA 1 and 2.

Induction of natural killer cell migration by monocyte chemotactic protein-1, -2 and -3.

Under certain physiological and pathological conditions, natural killer (NK) cells rapidly accumulate in tissues. Chemokines are an essential component of the current paradigm of leukocyte recruitment. The present study was designed to investigate the responsiveness of NK cells to the prototypic C-C chemokine, monocyte chemotactic protein-1 (MCP-1). MCP-1 induced migration across filters of interleukin (IL)-2-activated NK cells, whereas it was a weak attractant for unstimulated cells. Maximal induction of migration required a positive concentration gradient between the lower and the upper compartment of the chemotaxis chamber. Preliminary characterization of the MCP-1 receptor on NK cells indicated that the chemotactic response to MCP-1 was blocked by pre-treatment of cells with Bordetella pertussis toxin, and MCP-1 but not IL-8 displaced 125I-labeled MCP-1 from IL-2-activated NK cells. The related chemokines MCP-2 and MCP-3 were also active--though less potent--attractants for activated NK cells. Thus the spectrum of action of MCP-1, -2 and -3 encompasses NK cells and chemokines are likely to play a role in regulating extravasation of these cells.

Regulated expression of vascular cell adhesion molecule-1 in human malignant melanoma.

Expression of the endothelial adhesion molecule VCAM-1 was studied in human malignant melanoma lines by flow cytometry. Clones 2/4 and 2/14 (derived from the same lesion) had appreciable levels of VCAM-1 expression, whereas clone 2/21 and the lines A2058, Mel24, and A375 were negative. Clone 2/14 was selected for further analysis. Exposure to tumor necrosis factor (TNF) markedly augmented VCAM-1 on melanoma cells. Surface VCAM-1 was associated with expression of specific transcripts that were augmented by TNF. Analysis by reverse transcriptase and polymerase chain reaction using appropriate primers revealed that TNF-stimulated melanoma cells expressed both 7 and 6 immunoglobulin domain transcripts with predominance of the longer species. Tumor necrosis factor--stimulated melanoma cells bound more VLA-4-expressing cells (melanoma and monocytes) than resting tumor cells and anti-VCAM-1 monoclonal antibodies significantly inhibited binding, thus suggesting that surface VCAM-1 on melanoma is functional. Analysis of melanoma tissue sections demonstrated that VCAM-1 is not a marker of transformation of melanocytes because it can be detected in benign nevi. Although, unlike ICAM-1, VCAM-1 is not correlated with tumor progression, its expression in a fraction of primary melanomas indicates that it may play a role in regulating host immune response and homotypic interactions in some malignant melanomas.

Heterogeneous susceptibility of human melanoma clones to monocyte cytotoxicity: role of ICAM-1 defined by antibody blocking and gene transfer.

Five clones derived from the same human malignant melanoma lesion were studied for their susceptibility to killing by human monocytes activated by exposure to interferon (IFN)-gamma and lipopolysaccharide. Melanoma clones were heterogeneous in their susceptibility to human monocyte cytotoxicity, with one clone (2/21) exhibiting extremely low levels of lysis. The different levels of susceptibility to monocyte cytotoxicity were not accounted for by susceptibility or resistance to monokines [tumor necrosis factor (TNF), interleukin (IL)-1 and IL-6] because: (a) these effector molecules had little (TNF) or no (IL-1 and IL-6) cytolytic activity under these conditions; and (b) anti-TNF antibodies had marginal effects on cytotoxicity. Monocytes bound less to resistant than to susceptible melanoma cells. Monocyte-resistant 2/21 melanoma cells expressed substantially lower levels of ICAM-1 and VLA-4 than susceptible cells. Anti-CD18 and, to a lesser extent, anti-ICAM-1 mAb inhibited binding and cytotoxicity of human monocytes on malignant melanoma whereas anti-VLA-4 had no inhibitory action. Transfection of the ICAM-1 gene under the control of a constitutive promotor resulted in high levels of expression of ICAM-1 in 2/21 melanoma cells and, concomitantly, in augmented susceptibility to activated monocyte cytotoxicity. The augmented killing of ICAM-1 transfected 2/21 cells was inhibited by anti-ICAM-1 mAb. These results demonstrate that the CD18-ICAM-1 adhesion pathway can play an important role in the expression of human monocyte cytotoxicity on melanoma target cells and that heterogeneity in expression of ICAM-1 can underlie differences in susceptibility to tumoricidal activity.

Effect of acyclovir on selected immune functions.

The toxicity of acyclovir (ACV) produced by Lachema, Brno was compared with that of Zovirax, Wellcome. The in vitro suppressive effect of both substances was found equal and concentration dependent. The primary humoral antibody response was more sensitive to ACV than the cellular (blast transformation) response. However, spleen cells of drug-treated mice (either with the domestic compounds or Wellcome origin) differed neither in blast transformation test nor in the secretory antibody response. None of the drugs when given in adequate therapeutic dose did significantly influence the cell mediated response or antibody formation by spleen cells. Summing up, the acute immunotoxicity of both compounds was low; in this respect acyclovir Lachema did not differ from Zovirax Wellcome.

Molecules involved in the adhesion and cytotoxicity of activated monocytes on endothelial cells.

Our study was designed to investigate the surface molecules involved in the adhesion and cytotoxicity of activated human monocytes on resting and IL-1-stimulated endothelial cells (EC). Monocytes, exposed to the prototypic activating stimuli IFN-gamma and LPS, showed increased binding to resting and IL-1-treated EC. Activated monocytes were cytotoxic for resting and IL-1-treated EC in a 24- to 48-h [3H]TdR release assay. Anti-CD18 mAb significantly inhibited binding of monocytes on EC: in particular they caused 59 and 22% inhibition of adhesion of activated monocytes to resting and IL-1-stimulated EC, respectively. Anti-VLA4 mAb had little or no effect on binding when used alone, but combined use with anti-CD18 revealed an important role for this adhesion pathway: in particular, VLA4-dependent adhesion accounted for 40% of the binding of activated monocytes on IL-1-treated EC. Anti-CD18 mAb caused similar inhibition (77 and 81%) of the cytotoxicity of activated monocytes on resting and IL-1-treated EC in spite of the fact that this pathway accounted for only 22% of binding to activated EC. Moreover, anti-VLA4 mAb, alone or in combination with anti-CD18, had no effect on cytotoxicity. These results suggest that adhesion of activated monocytes to activated EC involves the CD18- and VLA4-dependent pathways, but that the former is dominant for the expression of cytotoxicity. Thus, in the ensemble of adhesion molecules available for interaction between endothelium and activated monocytes, the hierarchy of their importance may vary for different functions.

The signal transduction pathway involved in the migration induced by a monocyte chemotactic cytokine.

Recombinant monocyte-chemotactic and activating factor (rMCAF; alternative acronyms MCP-1, TDCF, human JE) induced migration of human monocytes across polycarbonate or nitrocellulose filters. Maximal induction of migration was observed at a concentration of 10 ng/ml (10(-9) M). Checkerboard analysis revealed that rMCAF elicited true gradient-dependent chemotactic migration, although a gradient independent chemokinetic effect was observed at low concentrations (1-5 ng/ml). rMCAF caused a rapid (less than 5 s) and transient (approximately 1.5 min) increase of free cytosolic Ca2+ ions, as assessed by the fura-2 probe. No Ca2+ increase was detected in neutrophils or lymphocytes stimulated by rMCAF. Studies conducted in the absence of extracellular Ca2+ or in the presence of Ni2+ (an inhibitor of Ca2+ influx) suggested that the increase of intracellular Ca2+ induced by rMCAF is dependent on the influx of extracellular Ca2+ through plasma membrane channels. Bordetella pertussis toxin inhibited the intracellular Ca2+ elevation and chemotaxis caused by rMCAF. The possible involvement of Ca(2+)-dependent protein kinases in rMCAF signaling pathway(s) was explored using inhibitors. Inhibitors of GMP-dependent kinase and myosin L chain kinase had no effect on rMCAF-induced monocyte migration. In contrast, protein kinase C/cAMP-dependent kinase inhibitors (such as, C-I, H-7, HA-1004, KT5720, and Staurosporine) markedly decreased rMCAF induced chemotaxis suggesting the involvement of a serine/threonine protein kinase, possibly protein kinase C, in rMCAF signaling pathway.

[Use of in vivo methods in the immunotoxicologic evaluation of drugs]. / Pouzití metod in vivo pro imunotoxikologické hodnocení léciv.

The immunity system is structurally and functionally multiform; in immunotoxicological screening therefore a larger number of manifestations of immunity response must be examined. Administration of drugs must be chosen in such a way as to enable studying the influence on the functions of immunity in the period prior to immunization, simultaneously with it, and after it. From this aspect the effect of subtoxic doses of cisplatinum, carboplatinum, oxoplatinum and iproplatinum administered to mice at different times with regard to the antigen was evaluated. In the case of antibody response (the cells forming antibodies in the spleen, serum haemagglutinins) as well as in the manifestations of cellular immunity (reaction of delayed hypersensitivity) the largest inhibition was registered on the administration from days 2 to 6 after the first contact with the antigen. Administration prior to immunization or simultaneously with it produced either mild stimulation or insignificant inhibition. The drugs most probably act by alkylating mechanism and the time course of the effect is similar to that in cyclophosphamide. The antibody response is inhibited most by oxoplatinum, cellular immunity by carboplatinum. When investigating phagocytic activity (Indian ink clearance from the blood circulation) only insignificant deviations were found. Immunity processes not requiring immediately cell division are substantially less influenced by platinum complexes.