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Biomedical subjects

J Sedlácek

Publications and source records attributed to J Sedlácek.

At least 19 recordsLinked to original sources
PubMed2005

[Comprehensive long-term analysis of hypospadias repair using vascularized flaps and tubularized incized plates--report on 588 cases].

BACKGROUND: The article presents a comprehensive long-term analysis of the modem hypospadias repair based on preservation and use of the urethral plate. METHODS AND RESULTS: In years 1987 to 2004, 476 patients (aged 8 months to 45 years) were operated using a praeputial (mostly) flap. 81% were indicated for reconstruction of a proximal form of hypospadias, mostly penile. In years 1986 to 1991, a tubularized flap was performed in 31 patients. Since 1992, an onlay island flap in 394 patients, inlay-onlay flap in 43 patients and onlay-tube-onlay flap in 8 patients have been used. In years 2000 to 2004, 112 patients (aged 13 months do 17 years) were operated using tubularized incised plate (TIP) repair, mostly because of distal form of hypospadias. To cover the neourethra, a dartos flap was used in 72 and spongioplasty in 37 patients. We achieved the primary healing in 89% and 91%, resp., of patients after onlay and onlay-inlay island flap repairs with an average follow-up of 6,7 and 5,7 years, respectively. The most frequent complications were fistula or dehiscence of the glans. After the TIP repair combined with a dartos flap, 89% of the first operated patients healed without complications with an average follow-up of 2.3 years. A higher complication rate (40%) was found after a spongioplasty with foreskin preservation. Secondary repairs were also associated with more complications. CONCLUSIONS: Using onlay and inlay-onlay one-stage repairs, or using TIP one-stage repair combined with a dartos flap, a successful primary hypospadias repair was achieved in 90% of patients. The definite repair should be completed during early childhood.

Adolescent↗
PubMed2005

[Effect of biofeedback in the treatment of voiding dysfunction].

BACKGROUND: Biofeedback represents a treatment option for children with voiding dysfunction characterised with poor pelvic floor relaxation. METHODS AND RESULTS: Thirty nine children with symptoms of voiding dysfunction were treated in BFB program. Average number of sessions to achieve good response was three. Follow up was 12 to 24 months. Very good clinical response was obtained in 55, 7% of patients, who were characterised by cessation or resolution of incontinence in 66, 7%, dyscoordination in 57, 6% and urinary tract infection in 41, 1% of children. CONCLUSIONS: Biofeedback therapy is an effective method for treating poor pelvic floor relaxation. It helps to improve coordination of the detrusor and external sphincter function and it accelerates voiding reflex maturation.

Adolescent↗
PubMed2003

[Benefits and risks of urologic laparoscopic surgery in adult patients].

The paper describes the advantages and disadvantages of the laparoscopic operations, the number of which steadily rises in urology. The laparoscopic surgery is considered to be a benefit regarding the short postoperative hospital stay, painless postoperative course, and virtually non-existing postoperative paralytic ileus. As disadvantage are deemed the long learning curve for the operating personal, and high economical costs, which could be cut down only if short off-work period in productive population is included. In the paper, the pathophysiological guidelines are outlined and emphasized during the laparoscopic operation, which the surgical and anesthesiological teams have to have in mind. On the own cohort of patients, the numbers and types of operations are described, which have been done at our department.

Adult↗
PubMed2001

A distinct binding mode of a hydroxyethylamine isostere inhibitor of HIV-1 protease.

Crystallization conditions for an HIV-1 protease-inhibitor complex were optimized to produce crystals suitable for X-ray diffraction experiments. The X-ray structure of the HIV-1 protease complex was solved and refined at 3.1 A resolution. In contrast to Saquinavir, the mimetic hydroxy group of the inhibitor Boc-Phe-Psi[(S)-CH(OH)CH(2)NH]-Phe-Glu-Phe-NH(2) is placed asymmetrically with respect to the non-crystallographic twofold axis of the protease dimer so that hydrogen bonds between the amino group of the inhibitor and the catalytic aspartates can be formed. The inhibitor binds in the centre of the active site by a compact network of hydrogen bonds to Gly27, Gly127, Asp25, Asp125 and via the buried water molecule W301 to Ile50 and Ile150.

Aspartic Acid↗
PubMed2001

[HPLC analysis of nonsteroidal antirheumatic substances in biological material. V. Fenamates].

The present paper surveys published HPLC methods estimating derivates of fenamic acid in biological materials and an HPLC method for estimation of meclofenamic acid in the samples of whole blood. Meclofenamic acid was isolated from the sample of the blood both liquid-liquid extraction and solid-liquid extraction. For determination of meclofenamic acid in the blood was preferred solid-liquid extraction. The adjust sample was analysed on reverse phase Separon SGX C-18 with the mobile phase methanol-water (pH 2.3) and detected at 220 nm. The method was used in a pharmacokinetic study of meclofenamic acid on rabbits.

Animals↗
PubMed2001

MRP 8/14 and procalcitonin serum levels in organ transplantations.

OBJECTIVES: MRP8/14 is a heterodimer of two myeloid calcium-binding proteins associated with different types of acute inflammatory processes. We studied MRP8/14 together with procalcitonin (PCT) serum levels in order to diagnose infectious complications or the rejection process affecting kidney or heart allograft. METHODS: A total of 419 serum samples was evaluated. MRP8/14 levels were measured by ELISA (BMA Biomed), PCT by a sensitive immunoluminiscent assay ILMA (Brahms Diagn.) RESULTS: Both parameters showed very low basal levels in healthy subjects (range 303-1,660 ng/ml of MRP8/14; less than 0.08 ng/ml of PCT). A rapid increase in serum levels occurred in response to bacterial infections (MRP8/14 up to 6,230 ng/ml; PCT up to 297 ng/ml). Serum PCT concentration remained low in the presence of kidney allograft rejection, where MRP8/14 levels were increased. An uncomplicated outcome of kidney or heart transplantation did not change basal serum MRP8/14 and PCT levels. CONCLUSIONS: We conclude that 1) both MRP8/14 and PCT are very sensitive markers of complications in organ transplant recipients (normal values in uncomplicated outcome) 2) combination of both parameters is useful to discriminate between rejection (increased MRP8/14 with normal PCT) and systemic bacterial infection (both parameters increased).

Aged↗
PubMed2000

Link between a novel human gammaD-crystallin allele and a unique cataract phenotype explained by protein crystallography.

We describe a 5-year-old boy with a unique congenital cataract caused by deposition of numerous birefringent, pleiochroic and macroscopically prismatic crystals. Crystal analysis with subsequent automatic Edman degradation and matrix-associated laser desorption ionization time-of-flight mass spectrometry have identified the crystal-forming protein as gammaD-crystallin (CRYGD) lacking the N-terminal methionine. Sequencing of the CRYGD gene has shown a heterozygous C-->A transversion in position 109 of the inferred cDNA (36R-->S transversion of the processed, N-terminal methionine-lacking CRYGD). The lens protein crystals were X-ray diffracting, and our crystal structure solution at 2.25 A suggests that mutant R36S CRYGD has an unaltered protein fold. In contrast, the observed crystal packing is possible only with the mutant protein molecules that lack the bulky Arg36 side chain. This is the first described case of human cataract caused by crystallization of a protein in the lens. It involves the third known mutation in the CRYGD gene but offers, for the first time, a causative explanation of the phenotype.

Alleles↗
PubMed2000

Cleavage of vimentin by different retroviral proteases.

Proteases (PRs) of retroviruses cleave viral polyproteins into their mature structural proteins and replication enzymes. Besides this essential role in the replication cycle of retroviruses, PRs also cleave a variety of host cell proteins. We have analyzed the in vitro cleavage of mouse vimentin by proteases of human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2), bovine leukemia virus (BLV), Mason-Pfizer monkey virus (M-PMV), myeloblastosis-associated virus (MAV), and two active-site mutants of MAV PR. Retroviral proteases display significant differences in specificity requirements. Here, we show a comparison of substrate specificities of several retroviral proteases on vimentin as a substrate. Vimentin was cleaved by all the proteases at different sites and with different rates. The results show that the physiologically important cellular protein vimentin can be degraded by different retroviral proteases.

Animals↗
PubMed2000

[HPLC analysis of nonsteroidal antirheumatic agents in biological materials. IV. Acetic acid derivatives].

The present paper surveys the published HPLC methods estimating derivatives of acetic acid in biological materials and an HPLC method for estimation of indomethacine in the samples of whole blood. The elaborated chromatographic conditions and process of isolation were applied also to the analysis of tropesine and both drugs were determined simultaneously. The blood samples were adjusted by liquid-liquid extraction and indomethacine (and tropesine) was analysed on the reverse phase Separon SGX C-18 with the mobile phase methanol-water (pH 3.2) and detected at 254 nm. The method was used in a pharmacokinetic study of indomethacine on rabbits.

Animals↗
PubMed1999

Serum procalcitonin concentrations in transplant patients with acute rejection and bacterial infections.

Procalcitonin (PCT) represents a new marker of systemic inflammatory reactions of the body to infections. PCT is selectively induced by severe bacterial infections leading to sepsis or multiorgan dysfunction syndrome. The aim of our study was to test PCT as a postoperative infection marker in heart and kidney transplant patients compared with healthy subjects and patients with localized lung-inflammatory processes without a manifest systemic response. PCT concentrations were measured by an immunoluminometric assay (ILMA) in a total of 419 serum samples. Normal serum levels were in the range of 0.08-0.6 ng/ml. Operative trauma associated with heart (not kidney) transplantation induced a transient increase in PCT levels to 7-10 ng/ml with a decline to normal levels within 2-3 days in most patients. Severe bacterial infections dramatically augmented serum PCT concentrations reaching values of 46-297 ng/ml in the most critical periods. Good response to antibiotic therapy was associated with a decline in serum PCT concentrations. Acute rejection or cytomegalovirus infections did not significantly increase the serum PCT levels. Localized pulmonary infections showed either no, or only a limited increase, in the serum PCT levels (max. 7 ng/ml). We conclude from our data that PCT can be used as a sensitive marker to differentiate systemic bacterial infections from other complications in organ transplantation.

Bacterial Infections↗
PubMed1999

Microcolumn high-performance liquid chromatographic assay for doxycycline in isolated alveolar macrophages.

A procedure for the isolation of doxycycline from the alveolar macrophages is described. Due to the minimal amount of the sample, it was necessary to employ microcolumn HPLC. A great advantage of the methodology developed in the present paper consists in the direct injection of an aqueous supernatant over disintegrated macrophages without the use of liquid-liquid or solid-liquid extraction. Another advantage of the method is the fact that the injection of an aqueous solution on a microcolumn with a lipophilic stationary phase makes the analyzed substances concentrate, which increases the sensitivity of assay. Separation was performed on a Separon SGX C18 microcolumn, 150 x 1 mm, detection at 254 nm by UV, 1 microliter flow cell. Rabbits were treated intravenously with Vibramycin inj. sic. in a dose of 13 mg/kg body weight doxycycline. A procedure for washing macrophages from rabbit lungs was developed. Dry matter of macrophages ranged from 5.9 to 11.4 mg. In the samples of three rabbits the levels of doxycycline ranged from 110 to 270 ng per 1 mg of macrophages dry matter.

Animals↗
PubMed1998

Effects of temperature and novobiocin on the expression of calf prochymosin gene and on plasmid copy number in recombinant Escherichia coli.

Escherichia coli strain HB101 harboring an expression plasmid bearing calf prochymosin gene under the control of the tac promoter was grown in the presence of IPTG with or without novobiocin at 28 and 40 degrees C, respectively. The differential rates of synthesis of prochymosin inclusions, and, for comparison, of beta-lactamase and beta-galactosidase, as well as plasmid copy number, were determined during the first hours of steady state growth. At 28 degrees C the induced expression of prochymosin gene was almost blocked. Addition of novobiocin did not alleviate this effect. In fact, it strengthened it, and we conclude that both these additive inhibitory effects are a consequence of the decrease in negative superhelical tension of plasmid DNA to an insufficient level. At 40 degrees C the differential rate of prochymosin synthesis was markedly enhanced. Since the copy number of the expression plasmid increased approximately to the same extent, we conclude that an increase in gene dose is the cause. The stimulation of cloned heterologous gene expression at 40 degrees C and inhibition at 28 degrees C may be conveniently used in biotechnological-scale cultivations of some recombinant bacteria.

Animals↗
PubMed1998

[Effect of acute exposure to SO2 on cellular defense mechanisms in the lung (pilot study)].

Sulphur dioxide is one of the most important and most frequent atmospheric pollutant of our environmental setting. In presented pilot-study we describe the influence of acute exposure to sulphur dioxide on alveolar macrophages of guinea pigs. The alveolar macrophages represent the main part of mononuclear phagocytic system, which is responsible for the defence of respiratory tract against foreign compounds. The experimental animals were exposed over three hours to sulphur dioxide with the concentration of about 400 ppm. After exposure we did not find significant changes in phagocytic activity of alveolar macrophages.

Animals↗
PubMed1998

[HPLC analysis of nonsteroidal antirheumatic agents in biological materials. III. Pyrazolidinediones].

The present paper surveys published HPLC methods estimating pyrazolidinediones in biological materials and an HPLC method for estimation of kebuzone in the samples of whole blood, isolated erythrocytes and plasma. An addition of propyl gallate prevented undesired oxidation of kebuzone. For liquid-liquid extraction, kebuzone was analysed on the reverse phases Separon SGX C-18 with the mobile phase methanol--water (pH 2.7) and detected at 247 nm. The elaborated method was used in a pharmacokinetic study of kebuzone on rabbits.

Animals↗
PubMed1998

On the intrinsic mechanisms of spontaneous motility in chick embryos.

Our experimental study was aimed on the problems of co-operation of NMDA- and NO-ergic mechanisms in the genesis of spontaneous motility in 17-day-old chick embryos. 1. Blockade of NO-synthase (NOS) by 7-nitro indazole prevented effect of an activation cocktail (NMDA + L-arginine). 2. Simultaneous blockade of NOS and of polyamine site in NMDA-receptor by arcaine was revealed by increased NMDA-activation of the spontaneous motility and by blockade of L-arginine and cocktail effects. 3. The blockade of NMDA-activation, depression of L-arginine activation and short-lasting potentiation of cocktail effect. 4. Blockade of NMDA mechanism by Mg2+ abolished activatory effect of both NMDA and L-arginine. The initial activation remained from the cocktail effect only. Results were considered as an evidence of a possible co-operation among NMDA- and NO-ergic mechanisms in the development of the embryonic spontaneous motility.

Animals↗
PubMed1997

Configurations of diastereomeric hydroxyethylene isosteres strongly affect biological activities of a series of specific inhibitors of human-immunodeficiency-virus proteinase.

Human immunodeficiency virus (HIV) proteinase (PR) represents an important target for antiviral chemotherapy. We present an analysis of inhibitory activities of a series of pseudopeptide inhibitors of HIV-1 PR. All inhibitors were N-protected tetrapeptides with the scissile bond replaced by a nonhydrolysable hydroxyethylene or hydroxyethylamine isostere. To elucidate subtle structural requirements of the PR binding cleft, we synthesised inhibitors with four combinations of configurations at the asymmetric carbons of the isostere. Compounds were tested in vitro using purified recombinant enzyme and a chromogenic peptide substrate. The differences in inhibition constants between individual diastereoisomers reached three orders of magnitude. The most active hydroxyethylene-containing inhibitor possessed the 2R,4S,5S configuration at the isostere. Inhibitor activity was also tested in mammalian cell culture by analysing reduction of viral polyprotein processing and virus infectivity. The results obtained in tissue culture were generally in agreement with the in vitro data, giving a similar order of potency for the individual diastereoisomers. The most active compounds completely blocked production of infectious virus. A simulation method for interaction was employed to build a model of the inhibitors in the PR active site, to identify the interactions responsible for the differences in activities of individual stereoisomers, and to estimate the relative contribution of individual structural features to the overall inhibitory activity.

Animals↗