Comparison of narrow-band imaging with autofluorescence imaging for endoscopic detection of squamous cell carcinoma of the tonsil.

PURPOSE: Early detection of mucosal neoplastic lesions is crucial for a patient's prognosis. This has led to the development of effective optical endoscopic diagnostic methods such as narrow band imaging (NBI) and autofluorescence (AFI). Independent of each other, both of these methods were proven useful in the detection of mucosal neoplasias. There are limited reported data comparing both methods for oropharyngeal cancer diagnostics. The aim of the study was to compare NBI and AFI endoscopic visualization of signs in identifying tonsillar squamous cell carcinoma (SCC) and assessing its extent and to determine whether the score was related to the evaluator's experience. METHODS: Patients with tonsillar SCC underwent endoscopic pharyngeal examination using NBI and AFI. Fiftyseven video sequences of examinations of lesions proven to be SCC were evaluated by three reviewers. The accuracy of determination of lesion extent and visualization of its endoscopic signs of malignancy were evaluated. RESULTS: Endoscopic visualization of tumour spread was significantly better using AFI than NBI (p = 0.0003). No significant difference was found between NBI and AFI in the visualization of endoscopic malignancy determining signs (p = 0.1405). No significant difference was found among the three reviewers in the visualization of tumour spread and for identifying malignancy-determining signs in NBI endoscopy or AFI endoscopy. CONCLUSIONS: The results show that AFI obtained better results for assessing the extent of tonsillar cancers than NBI. Both methods were proven to be equal in the visualization of endoscopic malignancy-determining signs. Both are useful even for less experienced evaluators.

Sedative effects of intranasal oxytocin in rabbits and rhesus monkeys.

Oxytocin is a hormone therapeutically used mainly for its peripheral effects during pregnancy in the uterus and breasts. However, additional central effects, i.e. anxiolytic effect, decreased level of social stress and increased empathy have been also observed. Hence, the aim of our study was to evaluate if nasal oxytocin can be used as anxiolytic substance in rhesus monkeys (n=20) and rabbits (n=20). Simultaneously, mean arterial blood pressure, arterial oxygen saturation of hemoglobin and pulse rate were monitored in all the evaluated animals. While rabbits lost righting reflex, monkeys developed a dose-dependent loss of aggressiveness and/or anxiety as evaluated by behavioral methods (aggressive behavior was classified as non-sedated - sedated - strongly sedated).

Ephedrine accelerates psychomotor recovery from anesthesia in macaque monkeys.

BACKGROUND: Ephedrine is used in treatment of hypotension during anesthesia. We investigated its effects on the psychomotor recovery and its potential adverse reactions on cardiorespiratory functions in rhesus monkeys. METHODS: The monkeys received 50 µg/kg medetomidine, 2.0 mg/kg S-ketamine with 150 IU hyaluronidase i.m. Pulse rate, blood pressure and saturation of haemoglobin were monitored for 20 minutes. Thereafter, 1 mg/kg of ephedrine or a placebo was administered i.m. and behavioural changes, pulse rate, blood pressure and saturation of haemoglobin were monitored every 5 minutes. RESULTS: Ephedrine shortened recovery from anaesthesia from 80.4 ± 25.8 to 14.83 ± 13.70 minutes. Ephedrine also increased oxygen saturation of haemoglobin and systolic blood pressure and caused significant decrease in pulse rate 5 minutes after its administration. CONCLUSIONS: Ephedrine can be successfully used to accelerate psychomotor recovery after the use of common anesthetic protocols combining dissociative anesthetic agent and alpha 2-adrenoceptor agonist in primates.

[Wound microbial sampling methods in surgical practice, imprint techniques]. / Metody odberu vzorku na kultivacní vysetrení v chirurgické praxi, otiskové metody.

The wound is a damage of tissue. The process of healing is influenced by many systemic and local factors. The most crucial and the most discussed local factor of wound healing is infection. Surgical site infection in the wound is caused by micro-organisms. This information is known for many years, however the conditions leading to an infection occurrence have not been sufficiently described yet. Correct sampling technique, correct storage, transportation, evaluation, and valid interpretation of these data are very important in clinical practice. There are many methods for microbiological sampling, but the best one has not been yet identified and validated. We aim to discuss the problem with the focus on the imprint technique.

[A combination of dexmedetomidine with ketamine and opioids results in significant inhibition of hemodynamic changes associated with laparoscopic cholecystectomy and in prolongation of postoperative analgesia]. / Kombinace dexmedetomidinu s ketaminem a opioidy významne potlacuje hemodynamické zmeny zpusobené laparoskopickou cholecystektomií a prodluzuje trváiní pooperacní analgezie.

THE AIM OF THE STUDY: Recently, alpha2 sympathoadrenergic drugs are used in premedication to improve the perioperative course. The aim of our study was to compare a premedication with a new alpha2 sympathoadrenergic drug and standard premedication. METHODS: After ethic committee approval and written patient consent, in a randomised, double-blinded study, combination of dexmedetomidine 1.0 microg x kg(-1) + ketamine 0.5 mg x kg(-1) + fentanyl 1.0 microg x kg(-1) + atropine 0.5 mg (group FNT), dexmedetomidine 1.0 microg x kg(-1) + ketamine 0.5 mg x kg(-1) + alfentanil 5.0 microg x kg(-1) + atropine 0.5 mg (group ALFNT), or pethidine 1.0 mg x kg(-1) + atropine 0.5 mg (group Dolsin) was administered to a deltoid muscle 15 min. before anaesthesia (GA) in patients elicited for laparoscopic cholecystectomy (LCHE). GA was performed in a standard way, ECG, NIBP, respiration rate, SpO2, onset of effect, Observers Assessment of Alertness Sedation Score (OAASS) before GA, circulatory reaction to intubation and capnoperitoneum, fentanyl consumption during GA, time to the first request for post-operative analgesia and postoperative nausea and vomiting were measured. The data were processed by Kruskal-Wallis and Fisher tests. P-value < 0.05 was considered significant. RESULTS: There were 16 patients in FNT and Dolsin and 15 patients in ALFNT with no differences in demography except for younger age in ALFNT. The main differences were in hypertension during capnoperitoneum: 0/16 FNT and 1/15 ALFNT vs. 11/16 Dolsin, both p < 0.001, per-operative fentanyl consumption: FNT 31.5 microg vs. Dolsin 165.0 microg, p < 0.001 and ALFNT 50.0 microg, p < 0.05 (ALFNT vs. Dolsin, p < 0.01) and request to the first analgesic post surgery: FNT 1.3 h. vs. Dolsin 0.45 h., p < 0.05 vs. ALFNT 0.8 h., p < 0.01. There were no differences in side effects except for bradycardia in ALFNT (p < 0.05). CONCLUSIONS: Dexmedetomidine-ketamine-fentanyl-atropine combination is superior to pethidine-atropine combination in suppressing of adverse hemodynamic effects of capnoperitoneum, decreased need for analgesia during GA and prolonged postoperative analgesia.

[Antibiotic resistance and biofilm formation in Staphylococcus saprophyticus strains isolated from urine]. / Staphylococcus saprophyticus--jeho rezistence k vybraným antibiotikum a tvorba biofilmu u kmenu izolovaných z moce.

Eighty-seven Staphylococcus saprophyticus strains isolated from urine of 87 patients with cystitis were examined in 2005-2009. All strains were tested for resistance to vancomycin, nitrofurantoin, doxycycline, oxacillin, amoxicillin/clavulanate, cefoxitin and sulfamethoxazole/trimethoprim and for biofilm formation by a modified Christensen method. None of the tested strains of S. saprophyticus showed resistance to vancomycin, while 2 strains (2.3 %) were resistant to nitrofurantoin, 9 (10.3%) to doxycycline, 20 (23.0 %) to oxacillin, 6 (6.9%) to amoxicillin/clavulanate, 6 (6.9%) to cefoxitin and 1 (1.1%) to sulfamethoxazole/trimethoprim. S. saprophyticus was detected as the causative agent of cystitis in 0.4 % of 20,375 culture positive urine samples analyzed in our laboratory between 2005 and 2009. Most 67 (77.0%) S. saprophyticus strains were recovered from women, particularly from young women. Biofilm formation was detected in 16 (18.4 %) out of 87 S. saprophyticus strains.

Experience with a naphthylmedetomidine-ketamine-hyaluronidase combination in inducing immobilization in anthropoid apes.

BACKGROUND: The aim of the study was to compare the effect of naphthylmedetomidine to medetomidine on the behavior of orangutans and chimpanzees. METHODS: The immobilization was performed as part of a medical examination in five chimpanzees and three orangutans. Following pre-medication with midazolam (0.70-1.20 mg/kg p.o.), naphthylmedetomidine (50-70 microg/kg), or medetomidine (20-30 microg/kg) was given with ketamine (3 mg/kg) and hyaluronidase (150 M.U.) into musculus deltoideus. RESULTS: We observed the distinct anti-aggressive effect of naphthylmedetomidine. The immobilization with naphthylmedetomidine was shallower and the influence on cardiac frequency less substantial compared to medetomidine. The overall sedative effect of naphthylmedetomidine lasted for less time, and its effect was incompletely antagonized with atipamezole in comparison to medetomidine. CONCLUSIONS: Naphthylmedetomidine could replace medetomidine for inducing immobilization and sedation. A combination of naphthylmedetomidine-ketamine is suitable for relocating animals to other cages or for painless medical examinations.

[Our experience in using fluorescence in situ hybridization FISH-Uro Vysion in diagnostics of urothelial carcinoma]. / Nase zkusenosti s metodou fluorescencní in situ hybridizace pri detekci uroteliálního karcinomu.

UNLABELLED: Urothelial carcinoma is a disease at high risk of recurrence after the initial therapy (70-80%) and with the tendency to progression accomplishing the recurrence (30%). Long lasting monitoring of patients with urothelial carcinoma is necessary. Cystoscopy and cytology are currently the primary modalities used to detect and monitor urothelial carcinoma. However, cytology has relatively poor sensitivity especially in well differentiated tumors. Cystoscopy is an invasive and relatively expensive method. Therefore, methods improving detection of urothelial carcinoma from urine specimens are employed. Uro Vysion (Vysis) fluorescence in situ hybridization (FISH) for improved detection of urothelial carcinoma was evaluated. MATERIALS AND METHODS: Bladder tumor progression is accompanied by increased chromosomal instability and aneuploidy of chromosomes 3, 7, 17 and loss of locus 9p21. A total of 124 patients were analyzed at Dpts. of Urology and Pathology, Faculty Hospital in Brno. Cytologically analyzed urine specimens were tested by FISH and simultaneously cystoscopy was employed including biopsy for histological examination. RESULTS: FISH analysis was positive in 35 cases, including 5 cases with negative biopsy and cytology. Negative FISH result was detected in 24 cases where the malignant status was determined. The sensitivity of FISH in our series was 58.9% and the specificity 88.1%. CONCLUSIONS: FISH is a relatively simple, speedy and non invasive diagnostic method. It detects the symptoms of malignity on the molecular level, which leads to earlier diagnosis and therapy and, hence, to potential extended survival. FISH makes it possible to take decision in cases of atypical or unclear cytological finding. The FISH method using the Uro Vysion kit appears as a prospective non invasive method capable of early UK detection, with a higher sensitivity than the standard cytology of urine.

[Comparison of susceptibility of spores of Bacillus subtilis and Czech strains of Clostridium difficile to disinfectants]. / Srovnání citlivosti spor Bacillus subtilis a spor ceských kmenu Clostridium difficile vuci dezinfekcním prostredkum.

An important factor in the prevention of nosocomial outbreaks caused by Clostridium difficile ribotype 027 is the disinfection of a patient environment by reliable sporicidal disinfectants. Sporicidal activity of particular agents is tested on spores of Bacillus subtilis. Questions are brought up if the disinfectant which works on B. subtilis spores will be equally effective on the spores of C. difficile. Therefore we have compared the effects of five disinfectants available on the Czech market on the spores of collection strains of both microbes and on the spores of ten C. difficile field strains isolated from feces of hospitalized patients. The effective substances were: disinfectant No. 1 chloramine B, No. 2 chlorine dioxide, No. 3 formaldehyde and ethan-2-dion, No. 4 peracetic and acetic acids and hydrogen peroxide, No. 5 ethanol and propan-2-ol. The testing was performed using the dilution neutralization method according to (SN EN 13704, the agent reducing the number of spores by more than 3 orders was considered sporicidal. In addition to the standard time 60 min a 15-minutes exposition was used and the effect was tested also under the protein burden. Disinfectant No. 1 showed better effect on the C. difficile than B. subtilis spores, even in lower (1%) concentration. Similarly, the sensitivity of the C. difficile spores to disinfectants No. 2 and 3 was somewhat higher. The sporicidity of the disinfectant No. 4 was so high that it reduced the number of spores of all strains within 15 minutes by more than 4 orders; possible difference in the susceptibility of spores was not observed. Whereas the disinfectant No. 5 was not reliably effective on the spores of B. subtilis, surprisingly it showed the sporicidal effect on the spores of field C. difficile strains. We conclude that spores of field C. difficile strains in particular turned out to be more sensitive to disinfectants than the spores of the collection strain ofB. subtilis. Therefore B. subtilis remains the right species for testing the sporicidal activity of disinfectants.

Antibiotic synergy against biofilm-forming Pseudomonas aeruginosa.

Eight antibiotics (aztreonam, ceftazidim, cefoperazon, cefepim, netilmicin, amikacin, ofloxacin and ciprofloxacin) exhibited antimicrobial activity individually and/or in combinations against 20 wild-type biofilm-forming strains of Pseudomonas aeruginosa. The strains were less susceptible in biofilm; in 10 strains antibiotic synergy was observed for the combination of aztreonam and ciprofloxacin. Synergy was also demonstrated in the case of beta-lactams and aminoglycosides, beta-lactams and fluoroquinolones, aminoglycosides and fluoroquinolones, and for monobactams and beta-lactams although the strains were resistant to the individual antibiotics. Synergism or partial synergism was found with one or more antibiotic combinations against 32.4% of isolates.

Importance of biofilm in Candida parapsilosis and evaluation of its susceptibility to antifungal agents by colorimetric method.

The ability of C. parapsilosis (an important cause of nosocomial infections) to produce biofilm was evaluated in 32 bloodstream isolates and 85 strains isolated from skin. The biofilm formation was found in 19 (59%) blood isolates and only in 33 (39%) isolates from skin. The antifungal susceptibility was assessed for amphotericin B, itraconazole and voriconazole in planktonic and biofilm form of the 19 biofilm-positive bloodstream strains by broth microdilution method according to NCCLS standards. The method was modified by the use of resazurin as a colorimetric indicator of the metabolically active cells which makes the determination of the effect of antifungal agents easier. Biofilm forms of all strains were more resistant than their planktonic form.

[Clinical specimens for PCR detection of syphilis]. / Klinický materiál pro prukaz syfilis metodou polymerázové retezové reakce.

A review is presented of types of clinical specimens used for diagnosis of syphilis by polymerase chain reaction. PCR is a routine method for detection of T. pallidum in swabs of chancres and primary and secondary syphilis mucocutaneous lesions. Whole blood, cerebrospinal fluid, amniotic fluid, aspirate and biopsy specimens, and paraffin-embedded tissues can also be tested by PCR for T. pallidum. However, further research on PCR detection sensitivity at various stages of syphilis is needed before these specimens are used in clinical practice.

[What are the prospects for medical microbiology in the next 20 years?]. / Co ceká lékarskou mikrobiologii za 20 let?

In the future, microbiological laboratories are expected to maintain their proficiency in both molecular biological procedures and conventional methods.

[Molecular detection and subtyping of Treponema pallidum subsp. pallidum in clinical specimens]. / Molekulární detekce a typizace Treponema pallidum subsp. pallidum v klinickém materiálu.

An in-house two-step nested PCR amplification targeting the tmpC gene (TP0319, encoding putative membrane lipoprotein) was used for detection of chromosomal DNA of Treponema pallidum subsp. pallidum in clinical specimens. We tested 138 blood serum samples from 111 adult patients with suspected, primary, secondary, early or late latent syphilis. T. p. pallidum DNA was not detected in any of the analyzed specimens. Out of 11 mucocutaneous swabs (7 genital and 4 pharyngeal), 6 collected from 3 patients with primary or secondary syphilis tested positive. One skin swab from a patient with early congenital syphilis was also positive as were his serum and cerebrospinal fluid samples. DNA sequencing of the genes TP0136 and TP0548 from the positive samples revealed two strains with DNA sequences identical to that of T. p. pallidum strain SS14 and two unique previously undescribed T. p. pallidum strains. The advances in molecular typing of T. p. pallidum in clinical specimens will be of relevance to the epidemiology of syphilis and will allow for clinical discrimination between reinfection and syphilitic reactivation.

[Detection and significance of biofilm formation in yeasts isolated from hemocultures]. / Detekce a význam tvorby biofilmu u kvasinek izolovaných z hemokultur.

BACKGROUND: Yeasts of the genus Candida are important opportunistic pathogens responsible for severe bloodstream infections in immunocompromised patients. An important virulence factor that allows them to colonize plastic implants and to survive in the blood is their ability to form biofilm. The purpose of the study was to assess, how frequently this virulence factor occurred in yeasts of the genus Candida isolated from blood cultures. MATERIALS AND METHODS: Cultures of the various strains in wells of microtitre plates were used to demonstrate the presence of biofilms. In total, we tested 72 isolates of yeasts of the genus Candida from the blood culture: C. albicans (33), C. parapsilosis (19), C. tropicalis (11), C. glabrata (4), C. krusei (3), C. lipolytica (1) and C. lusitaniae (1). RESULTS: The ability to form biofilm was demonstrated in 26 strains. The largest number of biofilm-positive strains was found in C. parapsilosis (13) and C. tropicalis (7). The formation of biofilms was far less frequent in the strains of C. albicans (4). The formation of biofilms was also demonstrated in C. krusei (2) and C. glabrata (1). CONCLUSION: The increasing frequency of fungal bloodstream infections is due to the increasing use of catheters and implants. The biofilm formation in these yeasts is the chief factor of virulence promoting such infections. The demonstration of biofilm formation in an isolated strain is indicative of the probable existence of a biofilm focus and of possible difficulties during antimycotic therapy.

[Methods for detection of biofilm formation in routine microbiological practice]. / Moznosti prukazu tvorby biofilmu v rutinní mikrobiologické praxi.

The increasing use of catheters, artificial implants and antimicrobials as well as high numbers of immunocompromised patients are major causes for concern over biofilm infections. These infections are characterized particularly by high resistance to antimicrobials and formation of persistent foci that may complicate therapy. Therefore, detection of biofilm formation is of high relevance to the clinician and his/her approach to the treatment. Reliable and sensitive methods for detection of this pathogenicity factor in clinically important organisms, suitable for use in routine microbiological laboratories, are needed for this purpose. Currently, a wide array of techniques are available for detection of this virulence factor, such as biofilm visualization by microscopy, culture detection, detection of particular components, detection of physical and chemical differences between biofilm-positive organisms and their planktonic forms and detection of genes responsible for biofilm formation. Since each of these methods has limitations, the best results can be achieved by combining different approaches.

[Microbiology online--interactive learning and a new discussion forum for microbiology teachers]. / Mikrobiologie on-line--výukové stránky a nové diskusní fórum pro ucitele mikrobiologie.

Rapid development of multiple disciplines of science, increasing numbers of university students and the advent of new learning technologies (e-learning, teleconferences, etc.) have drawn increasing interest in the contents, rationale and forms of learning. New Websites with a discussion forum for microbiology teachers have been available on the Web. In brief, the credo of this project called Forum is: "Teachers can learn from each other". The project is accessible free of charge at www.medmicro.info. Another source intended for free use by both teachers and students is "Learning pages of the Institute of Microbiology, Faculty of Medicine, Masaryk University" with the following sections: encyclopaedia, guidance on practical exercises, video clips and on-line laboratory using web cameras. The learning pages are available in both Czech and English.

Increased sensitivity to the acute effects of MDMA ("ecstasy") in female rats.

Behavioral effects of +/-3,4-methylenedioxymethamphetamine (MDMA, ecstasy) are relatively well described in humans as well as in animals. However, little is known about gender differences to the effects of MDMA. The aim of our study was to evaluate gender differences in stimulant effects of MDMA (2.5, 5.0, and 10.0 mg/kg subcutaneously (s.c.)) in male and female Wistar rats. We have used three behavioral methods (activity cage, open field, and elevated plus-maze) each describing a different pattern of spontaneous behavior. In the activity cage, 30 min after the MDMA administration, horizontal and vertical locomotor activities were registered for a period of 3 min. In the open field test rats were placed into an arena 15 min after drug treatment and locomotor activity was registered for a period of 30 min. Finally, in the elevated plus-maze test, rats were given MDMA 30 min prior to measurements and subsequently they were tested in the maze for a period of 5 min. In our experiments we observed a dose-dependent locomotion-enhancing effect of MDMA both in male and female rats in both locomotor tests. Female rats were more sensitive to the locomotor-stimulating effect than males in both tests, suggesting higher sensitivity to the stimulatory effect of MDMA. Further on, MDMA increased thigmotaxis in female rats in the open field test and decreased "anxious-like" behavior in the elevated plus-maze in both genders. In conclusion, we observed higher sensitivity of females to the locomotor-stimulant effect of MDMA. Increased sensitivity of females to the behavioral effects of MDMA can be explained by increased reactivity of serotonergic and dopaminergic systems.

[Czech terms for rod-shaped bacteria]. / Tyce, tycky, tycinky.

With increasing frequency, the rod-shaped bacteria have been recently referred to in Czech as tyce (the singular is tyc) instead of the diminutive tycinky (the singular is tycinka) as used in classical Czech textbooks and handbooks of medical microbiology. This discordance might result from inadequate translation of the English term rods. Although the English-Czech dictionary gives three equivalents for the analyzed meaning of this word, i.e. tyc, tycka, tycinka, the first option, a mono-syllable as the English rod is, is often selected without further consideration. Nevertheless, such simplification is not justified; to avoid confusion it is preferable to use conventional terminology.