Effects of yeast hydrolysate supplementation in low-fish meal diets for pikeperch.

Plant proteins have been increasingly used as sustainable substitutes for fish meal (FM) in aquafeeds; however, their high inclusion level compromises fish performance. The objective of this study was to examine whether yeast hydrolysate (YH) supplementation can improve the utilisation of high soybean meal (SM) diet and ameliorate its potential deteriorating impacts in pikeperch (Sander lucioperca). A basal diet was formulated using 44% FM, and four additional diets were produced by replacing 30 or 60% of FM with SM with or without the addition of 2% YH (FM, SM30, SM60, SM30 + YH, and SM60 + YH diets). Each diet was fed to three groups of fish (35.3 ± 0.10 g, 150 fish per group) to visual satiety four times daily for 70 days. Fish growth was not impacted by FM replacement level or YH application. However, SM60 group exhibited markedly higher feed conversion ratio and lower survival rate than those fed the FM- and YH-supplemented diets (P < 0.05). The highest and the lowest protein efficiency ratio values were obtained for the SM30 + YH and SM60 groups, respectively. Whole-body lipid content decreased in SM60 and SM60 + YH groups, and muscle lipid decreased in all the replacement groups. Serum triglyceride and glucose concentrations tended to decrease as FM replacement level increased. The highest alanine aminotransferase, aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities were detected in the SM60 group, and YH addition significantly decreased the AST and LDH activities. Serum lysozyme activity decreased in SM30, SM60 and SM60 + YH groups. Serum myeloperoxidase and antiprotease activities decreased in SM60 group, and YH supplementation improved their activities. No effects of diets were observed on serum antioxidant parameters such as catalase activity and malondialdehyde concentration, and gut morphological indices. Number of goblet cells in midgut decreased by increasing the SM inclusion level and a slight improvement was observed by YH application. These findings suggest that YH supplementation has the potential to support the replacement of up to 60% FM with defatted SM in pikeperch feed without deteriorating growth, feed utilisation, and survival rate. Further, YH incorporation mitigated the damaging impacts of high SM diet on liver function and non-specific immune response.

Tolerance of isolated rabbit hearts to short ischemic periods is affected by increased LV mass fraction.

Hypertrophied hearts are known for increased risk of arrhythmias and are linked with reduced ischemic tolerance. However, still little is known about state characterized only by increased left ventricle (LV) mass fraction. Seventeen isolated rabbit hearts with various LV mass were divided into two groups according to LV weight/heart weight ratio (LVW/HW ratio), namely group H and L (with higher and lower LVW/HW ratio, respectively) and underwent three short cycles of global ischemia and reperfusion. The differences in electrogram (heart rate, QRS(max), mean number, onset and dominant form of ventricular premature beats) and in biochemical markers of myocardial injury (creatine kinase, lactate dehydrogenase - LDH) and lipid peroxidation (4-hydroxy-2-nonenal - 4-HNE) were studied. As compared to group L, hearts in group H exhibited lower tolerance to ischemia expressed as higher incidence and severity of arrhythmias in the first ischemic period as well as increase of LDH and 4-HNE after the first reperfusion. In the third cycle of ischemia-reperfusion, the preconditioning effect was observed in both electrophysiological parameters and LDH release in group H. Our results showed consistent trends when comparing changes in electrograms and biochemical markers. Moreover, 4-HNE seems to be good potential parameter of moderate membrane alteration following ischemia-reperfusion injury.

A B-cell epigenetic signature defines three biologic subgroups of chronic lymphocytic leukemia with clinical impact.

Prospective identification of patients with chronic lymphocytic leukemia (CLL) destined to progress would greatly facilitate their clinical management. Recently, whole-genome DNA methylation analyses identified three clinicobiologic CLL subgroups with an epigenetic signature related to different normal B-cell counterparts. Here, we developed a clinically applicable method to identify these subgroups and to study their clinical relevance. Using a support vector machine approach, we built a prediction model using five epigenetic biomarkers that was able to classify CLL patients accurately into the three subgroups, namely naive B-cell-like, intermediate and memory B-cell-like CLL. DNA methylation was quantified by highly reproducible bisulfite pyrosequencing assays in two independent CLL series. In the initial series (n=211), the three subgroups showed differential levels of IGHV (immunoglobulin heavy-chain locus) mutation (P<0.001) and VH usage (P<0.03), as well as different clinical features and outcome in terms of time to first treatment (TTT) and overall survival (P<0.001). A multivariate Cox model showed that epigenetic classification was the strongest predictor of TTT (P<0.001) along with Binet stage (P<0.001). These findings were corroborated in a validation series (n=97). In this study, we developed a simple and robust method using epigenetic biomarkers to categorize CLLs into three subgroups with different clinicobiologic features and outcome.

Motion artefact in voltage-sensitive fluorescent dye emission during repeated ischemia of isolated heart.

Motion artefact (MA) in voltage-sensitive fluorescent signals causes significant debasement of action potential. During ischemia and reperfusion in isolated rabbit heart, this artefact develops in a manner which may be described by the time of its onset, level, and shape. The MA during ischemia: (a) may become substantial with approximately two minutes delay after establishing global ischemia; (b) may be almost twice as high as the physiological action potential and decreases both with time and repetition of ischemia; (c) the MA shape is unpredictable and depends on individual rabbit.

Application of the optical method in experimental cardiology: action potential and intracellular calcium concentration measurement.

It has been shown that, in addition to conventional contact electrode techniques, optical methods using fluorescent dyes can be successfully used for cardiac signal measurement. In this review, the physical and technical fundamentals of the method are described, as well as the properties of the most common systems for measuring action potentials and intracellular calcium concentration. Special attention is paid to summarizing limitations and trends in developing this method.

Three-dimensional electrogram in spherical coordinates: application to ischemia analysis.

Three-dimensional electrogram was used for analysis of ischemia manifestation in isolated hearts. Three parameters based on spherical coordination system were used in this study - amplitude of electrical heart vector, its azimuth and elevation. The parameters were presented as a trend. This approach reflected ischemic changes in a manner which can be easily observed and evaluated. Ischemia was analysed in seven isolated hearts of New Zealand white rabbits. It was found that (a) ischemia changes heart electric vector, (b) ischemic preconditioning has a protective effect, and (c) both of these findings can be clearly observed by the proposed method.

Experimental methods for simultaneous measurement of action potentials and electrograms in isolated heart.

Monophasic action potential (MAP) can be recorded from the heart surface by optical method based on fluorescence measurement. The motion of isolated heart during experiment caused additional noise in recorded signal. The motion artifact can be eliminated by ratiometric fluorescence emission measurements. This study is based on experiments in which optical MAP measurement is done by single-wavelength and dual-wavelength measurement of fluorescence emission. Both recording setups are presented and their advantages and disadvantages are discussed. MAPs recorded by both methods from isolated rabbit hearts perfused according to Langendorff are presented. Simultaneous electrograms (EG) and MAPs recording are analyzed and measurement of velocity of impulse conduction through heart tissue is presented.

Sex differentiation and vitellogenin and 11-ketotestosterone levels in chub, Leuciscus cephalus L., exposed to 17 beta-estradiol and testosterone during early development.

The effects of 17 beta-estradiol (E(2)) and testosterone (T) singly and in combination were tested on juvenile chub (Leuciscus cephalus L.). Vitellogenin (VTG) and 11-ketotestosterone (11-KT) were determined by ELISA in whole body homogenates and the gonads were examined histologically. Testosterone and estradiol, in combination, significantly increased whole body VTG (p < 0.01), but not 11-KT, compared to controls and the T treated groups. The only intersex observed (1/80) was in the combined treatment group. We suggest that VTG measured in whole body homogenates could be used to determine the effects of exogenous steroids in juvenile chub.

Effects of pollution on chub in the River Elbe, Czech Republic.

The Elbe River is one of the most polluted aquatic ecosystems in the Czech Republic. The effect of three major chemical plants located on the Elbe River (at Pardubice, Neratovice, and Usti nad Labem) on fish was studied in 2004. Health status, chemical concentrations (Hg, PCB, DDT, HCH, HCB, OCS, 4-tert-nonylphenols, 4-tert-octylphenol) in muscle, and biomarkers (hepatic ethoxyresorufin-O-deethylase (EROD), plasma vitellogenin, and plasma 11-ketotestosterone) were assessed in male chub (Leuciscus cephalus L.). Differences between localities upstream (US) and downstream (DS) from the monitored source of pollution were identified. Fish from DS sampling sites showed significantly higher levels of contaminants than fish from US sampling sites. Generally, the concentrations of pollutants in fish from the Elbe sites were significantly higher compared to the reference site. Reduced gonad size, decreased plasma levels of 11-ketotestosterone, EROD and vitellogenin induction, and histopathologies of male gonads indicated harmful effects of aquatic pollution in fish from the Czech portion of the Elbe River.

[Importance of genetic testing in couples with reproductive disorders]. / Význam genetického vysetrení u páru s poruchami reprodukce.

OBJECTIVE: To determine the prevalence of chromosomal aberrations in infertile couples undergoing in vitro fertilization (IVF). DESIGN: Cytogenetic analysis of peripheral blood lymphocytes in the group of patients undergoing IVF. Detection of chromosomal aberrations in the fetuses after IVF. SETTING: Department of Medical Genetics and Fetal Medicine, Medical Faculty, Palacký University and the University Hospital, Olomouc. METHODS: Cultivation of peripheral blood lymphocytes or fibroblasts of amniotic fluid. Using fluorescent in situ hybridization in cases of mosaicism. RESULTS: Out of 638 patients undergoing treatment for male or female infertility, 595 had normal karyotype and 43 (6.8%) had abnormal karyotype. There were detected 9 (1.4%) cases of balanced chromosomal rearrangements, 2 (0.31%) cases of deletion of Y chromosome, 2 (0.31%) cases of inversion, 2 (0.31%) cases of marker chromosome, 5 (0.78%) cases of gonosomal aneuploidy (47,XXY) and 23 (3.65%) cases of gonosomal mosaicism--out of the 22 (3.5%) cases of low-level mosaicism. In the small group of pregnant patients after IVF investigated for the risk of genetic disorders included in our study (n = 60) the frequency of chromosomal abnormalities was 9 (15%). CONCLUSIONS: Our data show that a high number of infertile couples is affected by chromosomal aberrations which occur more frequently in females than in males. It is caused by high frequency of low-level gonosomal mosaicism in the group of infertile women. Chromosomal analyses are highly recommended before each IVF procedure.

[Microdeletion of the azoospermia factor as one of the causes of male infertility]. / Mikrodelece faktoru azoospermie jako jedna z prícin muzské infertility.

One of the possible causes of male infertility is microdeletion of the Y chromosome in the Yq11.23 region--named the azoospermia factor. These deletions are associated with azoospermia or severe oligozoospermia. In these cases, testicular histopathological findings comprise a wide spectrum, from total absence of germ cells, through arrest of their maturation to decreased sperm production. Most Y-chromosome microdeletions arise de novo but transmission from the father is also possible, either by the natural way or by assisted reproduction. In relation to the assisted reproduction, the relationship between the Y-deletions and presence of spermatozoa in testis, fertilization capability and embryo quality were examined. Heredity of the deleted Y chromosome is holandric and therefore all sons of males with deletions will carry the same defect and will probably have fertility problems. Another negative influence of deletions on a man's health has not yet been identified.

[Analysis of specific sequences in female patients with Turner syndrome--initial study]. / Analýza y specifických sekvencí u pacientek s turnerovým syndromem--úvodní studie.

BACKGROUND: DNA sequences from chromosome Y can cause gonadoblastoma development in patients with Turner syndrome (TS). Estimated risk is about 30%. The aim of the study is detection of Y-sequences of DNA level, calculation of mosaicism and its cytogenetic location. Clinical result of the study is the recommendation to gonadectomy of proved positive patients. METHODS AND RESULTS: Samples from 110 patients were collected. The PCR method and analysis of products on agarose gel was compared with analysis of DNA fragments from quantitative fluorescent (QF) PCR on capillary electrophoresis. The loci DYZ3, AMGX/Y and SRY were used for detection. The method QF PCR was effected for DYZ3 and AMGX/Y loci. The positive cases were examined by FISH method. Five (4.5%) and 3 (2.7%) positive cases were detected in DYZ3 and SRY resp. loci by electrophoresis on agarose gel. Seventeen (15.5%) and 7 (6.4%) positive cases were detected in DYZ3 and AMGX/Y resp. by capillary electrophoresis. The estimated mosaicism ranged from 1:5 to 1:100,000. CONCLUSIONS: QG PCR is the most sensitive method for diagnostics of Y-sequences. Simultaneously the incidence of Y-positive cells can be estimated. The positive cases with marker in karyotype were confirmed by FISH.

[DNA analysis of chromosome Y in the area of the azoospermia factor (AZF) in infertile men]. / DNA analýza chromozomu Y v oblasti AZF u sterilních muzu.

OBJECTIVE: Establishment of investigation of sterile male DNA in AZF region--choice of loci and primers for investigation, optimization of PCR conditions (polymerase chain reaction). DESIGN: For practice. SETTING: Department of Medical Genetics and Foetal Medicine, Faculty of Medicine, Palacky University and Faculty Hospital Olomouc. METHODS: PCR amplification of DNA isolated from blood of sterile men and consequential electrophoresis of synthesized DNA fragments to appoint microdeletions in AZF. RESULTS: From January to June 2000 were detected the microdeletions in AZF of 3 out of 79 sterile men (3.80%) by means of the Experteam firm kit. From July to December 2000 were tested and optimized conditions of amplification of 10 AZF loci to substitute the kit and they were used for examination of the first 20 sterile men of our collection. CONCLUSION: In our laboratory was established routine examination male sterility related to microdeletions in AZF. With our collection of loci was substituted the original Experteam firm kit and was widened spectrum of observed loci.

Successful ventricular defibrillation by the selective sodium-hydrogen exchanger isoform-1 inhibitor cariporide.

BACKGROUND: Sodium-hydrogen exchanger isoform-1 (NHE-1) activation worsens functional myocardial abnormalities associated with ischemia and reperfusion. We hypothesize that these abnormalities may limit cardiac resuscitation from ventricular fibrillation (VF) and investigated whether NHE-1 inhibition with the benzoylguanidine derivative cariporide could improve resuscitability, postresuscitation myocardial function, and short-term survival in isolated heart and intact rat models of VF. Methods and Results-- In the isolated rat heart, VF was induced for 25 minutes. Perfusion was interrupted for the initial 10 minutes and restarted at 10% of baseline flow for the remaining 15 minutes (simulating chest compression). Cariporide ameliorated ischemic contracture, prevented postresuscitation diastolic dysfunction, and favored earlier return of contractile function. In the intact rat, cariporide, injected into the right atrium before chest compression was started (after 6 minutes of untreated VF), prompted spontaneous defibrillation between minutes 7 and 9 of chest compression in 6 of 8 rats. In contrast, electrical defibrillation was required in each of 8 control rats after completion of a predetermined 16-minute interval of VF. After resuscitation, cariporide-treated rats had less ventricular ectopic activity and normalized their hemodynamic function faster. Electrical defibrillation was then timed in control rats to match the time when spontaneous defibrillation occurred in cariporide-treated rats. With comparable VF duration, postresuscitation hemodynamic dysfunction was ameliorated by cariporide, but only when more severe ischemia was modeled by prolongation of the interval of untreated VF from 6 to 10 minutes. CONCLUSION: NHE-1 inhibition may represent a novel and remarkably effective intervention for resuscitation from VF.

[Cytotoxicity tests of stomatologic materials on cell cultures]. / Testování cytotoxicity stomatologických materiálu na bunecných kulturách.

Authors compare the benefits and drawbacks of agar diffusion tests (with neutral red and with thiazolyl blue) and the filter diffusion test recommended as the standards for cytotoxicity testing of dental materials and their eluates. It seems that there are no differences between these diffusion tests after the treatment for 24 hours. Authors discuss the observed differences in the toxicity between a solid form of the material (ANA 2000 DUO, Evicrol Solar LC) and its eluate.

Biochemical markers for differentiation of exposures to nonplanar polychlorinated biphenyls, organochlorine pesticides, or 2,3,7, 8-tetrachlorodibenzo-p-dioxin in trout liver.

The effects of a single intraperitoneal dose of the prototypical contaminant nonplanar 2,2',4,4',5,5'-hexachlorobiphenyl (PCB 153, 50 mg/kg), p,p'-DDE (50 mg/kg), or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 200 ng/kg) on the activities of hepatic detoxification enzymes were examined in the liver of immature rainbow trout (Oncorhynchus mykiss). Different modulations of the tested xenobiotics on microsomal cytochrome P450-dependent testosterone hydroxylase activities were found: PCB 153 specifically induced 16beta-hydroxylase activity, whereas p,p'-DDE decreased cytochrome P4503A-dependent 6beta-hydroxylation as well as 16alpha- and 2alpha-hydroxylation. TCDD did not modulate testosterone hydroxylase activities, but a strong induction of cytochrome P4501A activity was observed after TCDD administration; hence, cytochrome P4501A is not involved in the hydroxylation of testosterone. Trout hepatic microsomal glutathione S-transferase (GST) activity, enhanced by all the xenobiotics tested, was found to be a sensitive nonspecific biochemical marker of oxidative stress; cytosolic glutathione reductase was a less sensitive indicator of oxidative stress and was induced significantly only by treatment with p,p'-DDE. Cytosolic GST activity toward ethacrynic acid (GST-ETHA) was induced by PCB 153 or p,p'-DDE, but not by TCDD. Modulations of hepatic microsomal testosterone hydroxylase activities and induction of GST-ETHA appeared to be suitable biochemical markers of acute exposure to nonplanar PCBs and organochlorines that do not induce cytochrome P4501A enzymes in rainbow trout, whereas microsomal GST and cytosolic glutathione reductase may become early biochemical indicators of oxidative stress.

[Preparation trauma in stomatology]. / Preparacní trauma ve stomatologii.

In this paper authors deal with the causes of preparation trauma in stomatology. They have studied effects of high temperature on human cells cultured in vitro. Based both on literature data and on their own experience they summarize basic principles of preparation which prevent preparation trauma. They summarize how to eliminate as much as possible factors that damage hard dental tissues and pulp.

[Physical, chemical and biological aspects of treatment of pathological changes in hard dental tissues]. / Fyzikální, chemická a biologická hlediska osetrení patologických zmen tvrdých zubních tkání.

The methods of preparation of dental tissues, treatment of the dentine wound and filling materials, which replace the hard dental tissues, are discussed to evaluate their features in toward the requirements for biological harmlessness. Based on the literature data, their own clinical and practical experiences, the authors have found that none of the currently used preparation methods as well as supports or filling materials are fulfilling the biological treatment requirements. Therefore, it is necessary to be aware of this fact, and adjust the medical procedures in such a way to avoid the harmful effects or to suppress then as much as possible until the ideal preparation procedures and filling materials will be developed.

[Biological aspects of materials recommended as alternatives to amalgams]. / Biologické aspekty materiálu doporucovaných jako alternativy za amalgám.

Evaluation of biological properties of filling materials used as an alternative to amalgams shows that none of the tested materials possess the required biological properties. It is therefore necessary to search for new and better filling materials.