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Background: Current screening methods of colorectal carcinoma are based on examination of occult bleeding in the stool, and further on endoscopic and irrigographical (barium enema) examinations. Population-based non-invasive screening method having high sensitivity and specificity is needed., Methods: Detection of molecular alterations in colonocytes from the stool may be a promising new diagnostic tool for such screening. Determination of mutations in APC, K-ras, DCC, p53 genes and "long" DNA may serve for early detection of colorectal cancer from stool samples. Multi-target DNA-assays employing all these markers suggest high sensitivity and specificity, unfortunately also expensiveness. Therefore finding a marker characteristic for all tumor cells would be desirable. Nuclear faktor-kappaB (NF-kappaB) could be such marker suitable for determination in colonocytes shed into the stool., Conclusion: Molecular testing of stool for early detection of colorectal cancer may be a promising screening method for this disease. Large multicenter trials are required to validate results obtained from preliminary clinical studies.

The determination of genetic variability by means of molecular genetic markers FUT1 and ESR2, and the evaluation of their associations with reproduction traits in population of Prestice Black-Pied sows have been implemented. A lower frequency of the allele FUT1A than the frequency of the allele FUT1G in the FUT1 gene was ascertained. In the ESR2 polymorphism, a higher frequency of the allele ESR2A compared with the allele ESR2B was determined. In the second to sixth litters as well as in the first to sixth litters, a significantly lower number of piglets born alive and number of weaned piglets from sows of the FUT1/IFUT1A genotype was found. Moreover, the homozygotes FUT1G /FUT1G significantly exceeded sows of the FUT1A/FUT1A genotype in the total number of born piglets in the first to sixth litters. No significant effect of the ESR2 polymorphism on reproduction was observed.

The retrospective case review was aimed to demonstrate anatomical and functional results of pars plana vitrectomy (PPV) in patients with Terson's syndrome (TS). The most common cause of TS was an acute subarachnoid hemorrhage of ruptured intracranial aneurysm (6 eyes). One patient suffered traumatic subdural and subarachnoid hemorrhage. PPV was performed in 7 eyes of 6 patients (2 women and 4 men). The patients ranged in age from 18 to 53 years (average 37.5 year), the mean age was 37.5 years. The interval between intracranial hemorrhage and PPV varied from 2 to 12 month (average 7.5 months). The visual acuity postoperatively improved in all 7 eyes. The mean period of observation was 12.5 month. Intraoperative complications included retinal break (1) only. Late complications included epiretinal membrane (2), glaucoma (1), cataract (2) and conjunctival cyst (1). Pars plana vitrectomy is highly effective and relatively safe method in hastening visual rehabilitation of adults with Terson's syndrome.

Retinopathy of prematurity (ROP) is a proliferative retinopathy affecting predominantly immature children and children with low birth weight under 2500 g in 5-6%. During 11 years we operated on 15 eyes of 15 patients aged 5-31 years, because of late tractional retinal detachment (TRD) appearing on the basis of ROP. The average follow-up period was 37.2 months; the minimal follow-up period was 1 year. In all patients, the personal medical history revealed the pre-term birth at the gestation age of 28-34 weeks. The birth weight was in the range 850-1600 grams. According to the extend of vitreoretinal traction and type of retinal defect we indicated: a) primary cryosurgical procedure (CP) with episcleral silicone implants. The operation was indicated in 6 eyes: cerclage, segmental perilimbal plombage in the temporal half and puncture of the subretinal fluid; b) pars plana vitrectomy (PPV). In total in 9 eyes the PPV was indicated, in 5 eyes it was supplemented with relaxing retinotomy, and in 7 eyes with silicone oil (SO) implantation. Three eyes were re-operated by means of PPV with SO implantation. The final anatomical success of the surgical treatment of the prognostic unfavorable TRD, including re-operations was achieved in 12 eyes (80%), which we consider as a good one. The visual acuity improved or remained the same also in 12 eyes (80%) and visual acuity 6/60 and better (20/200 or 0.1) in 8 out of 15 eyes (53%).

A novel and quick high-performance liquid chromatography (HPLC) method with UV spectrophotometric detection was developed and validated for the determination of five compounds in topical gel. The described method is suitable for simultaneous determination of active component ketoprofen, two preservatives methylparaben and propylparaben and two degradation products of ketoprofen--3-acetylbenzophenone and 2-(3-carboxyphenyl) propionic acid--in a topical cream after long-term stability tests using ethylparaben as an internal standard. The chromatographic separation was performed on a 5microm Supelco Discovery C18 column (125mm x 4mm i.d., Sigma-Aldrich); the optimal mobile phase for separation of ketoprofen, methylparaben, propylparaben, degradation products 3-acetylbenzophenone and 2-(3-carboxyphenyl) propionic acid and ethylparaben as internal standard consists of a mixture of acetonitril, water and phosphate buffer pH 3.5 (40:58:2, v/v/v). At a flow rate of 1.0ml min(-1) and detection at 233nm, the total time of analysis was less than 10min. The method was applied for routine analysis (batch analysis and stability tests) of these compounds in topical pharmaceutical product.

To localize wheat (Triticum aestivum L.) ESTs on chromosomes, 882 homoeologous group 6-specific ESTs were identified by physically mapping 7965 singletons from 37 cDNA libraries on 146 chromosome, arm, and sub-arm aneuploid and deletion stocks. The 882 ESTs were physically mapped to 25 regions (bins) flanked by 23 deletion breakpoints. Of the 5154 restriction fragments detected by 882 ESTs, 2043 (loci) were localized to group 6 chromosomes and 806 were mapped on other chromosome groups. The number of loci mapped was greatest on chromosome 6B and least on 6D. The 264 ESTs that detected orthologous loci on all three homoeologs using one restriction enzyme were used to construct a consensus physical map. The physical distribution of ESTs was uneven on chromosomes with a tendency toward higher densities in the distal halves of chromosome arms. About 43% of the wheat group 6 ESTs identified rice homologs upon comparisons of genome sequences. Fifty-eight percent of these ESTs were present on rice chromosome 2 and the remaining were on other rice chromosomes. Even within the group 6 bins, rice chromosomal blocks identified by 1-6 wheat ESTs were homologous to up to 11 rice chromosomes. These rice-block contigs were used to resolve the order of wheat ESTs within each bin.

Common wheat (Triticum aestivum L.,2n=6x=42) is an allohexaploid composed of three closely related genomes, designated A, B, and D. Genetic analysis in wheat is complicated, as most genes are present in triplicated sets located in the same chromosomal regions of homoeologous chromosomes. The goal of this report was to use genomic information gathered from wheat-rice sequence comparison to develop genome-specific primer sets for five genes involved in starch biosynthesis. Intron locations in wheat were inferred through the alignment of wheat cDNA sequences with rice genomic sequence.Exon-anchored primers, which amplify across introns,allowed the sequencing of introns from the three genomes for each gene. Sequence variation within introns among the three wheat genomes provided the basis for genome-specific primer design. For three genes, ADP-glucose pyrophosphorylase (Agp-L), sucrose transporter (SUT),and waxy (Wx), genome-specific primer sets were developed for all three genomes. Genome-specific primers were developed for two of the three genomes for Agp-S and starch synthase I (Ssl). Thus, 13 of 15 possible genome-specific primer sets were developed using this strategy. Seven genome-specific primer combinations were used to amplify alleles in hexaploid wheat lines for sequence comparison. Three single nucleotide polymorphisms(SNPs) were identified in a comparison of 5,093 bp among a minimum of ten wheat accessions. Two of theseSNPs could be converted into cleaved amplified polymorphism sequence (CAPS) markers. Our results indicated that the design of genome-specific primer sets using intron-based sequence differences has a high probability of success, while the identification of polymorphism among alleles within a genome may be a challenge.

This report describes the rationale, approaches, organization, and resource development leading to a large-scale deletion bin map of the hexaploid (2n = 6x = 42) wheat genome (Triticum aestivum L.). Accompanying reports in this issue detail results from chromosome bin-mapping of expressed sequence tags (ESTs) representing genes onto the seven homoeologous chromosome groups and a global analysis of the entire mapped wheat EST data set. Among the resources developed were the first extensive public wheat EST collection (113,220 ESTs). Described are protocols for sequencing, sequence processing, EST nomenclature, and the assembly of ESTs into contigs. These contigs plus singletons (unassembled ESTs) were used for selection of distinct sequence motif unigenes. Selected ESTs were rearrayed, validated by 5' and 3' sequencing, and amplified for probing a series of wheat aneuploid and deletion stocks. Images and data for all Southern hybridizations were deposited in databases and were used by the coordinators for each of the seven homoeologous chromosome groups to validate the mapping results. Results from this project have established the foundation for future developments in wheat genomics.

Because of the huge size of the common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) genome of 17,300 Mb, sequencing and mapping of the expressed portion is a logical first step for gene discovery. Here we report mapping of 7104 expressed sequence tag (EST) unigenes by Southern hybridization into a chromosome bin map using a set of wheat aneuploids and deletion stocks. Each EST detected a mean of 4.8 restriction fragments and 2.8 loci. More loci were mapped in the B genome (5774) than in the A (5173) or D (5146) genomes. The EST density was significantly higher for the D genome than for the A or B. In general, EST density increased relative to the physical distance from the centromere. The majority of EST-dense regions are in the distal parts of chromosomes. Most of the agronomically important genes are located in EST-dense regions. The chromosome bin map of ESTs is a unique resource for SNP analysis, comparative mapping, structural and functional analysis, and polyploid evolution, as well as providing a framework for constructing a sequence-ready, BAC-contig map of the wheat genome.

The objectives of this study were to develop a high-density chromosome bin map of homoeologous group 7 in hexaploid wheat (Triticum aestivum L.), to identify gene distribution in these chromosomes, and to perform comparative studies of wheat with rice and barley. We mapped 2148 loci from 919 EST clones onto group 7 chromosomes of wheat. In the majority of cases the numbers of loci were significantly lower in the centromeric regions and tended to increase in the distal regions. The level of duplicated loci in this group was 24% with most of these loci being localized toward the distal regions. One hundred nineteen EST probes that hybridized to three fragments and mapped to the three group 7 chromosomes were designated landmark probes and were used to construct a consensus homoeologous group 7 map. An additional 49 probes that mapped to 7AS, 7DS, and the ancestral translocated segment involving 7BS also were designated landmarks. Landmark probe orders and comparative maps of wheat, rice, and barley were produced on the basis of corresponding rice BAC/PAC and genetic markers that mapped on chromosomes 6 and 8 of rice. Identification of landmark ESTs and development of consensus maps may provide a framework of conserved coding regions predating the evolution of wheat genomes.

A total of 37 original cDNA libraries and 9 derivative libraries enriched for rare sequences were produced from Chinese Spring wheat (Triticum aestivum L.), five other hexaploid wheat genotypes (Cheyenne, Brevor, TAM W101, BH1146, Butte 86), tetraploid durum wheat (T. turgidum L.), diploid wheat (T. monococcum L.), and two other diploid members of the grass tribe Triticeae (Aegilops speltoides Tausch and Secale cereale L.). The emphasis in the choice of plant materials for library construction was reproductive development subjected to environmental factors that ultimately affect grain quality and yield, but roots and other tissues were also included. Partial cDNA expressed sequence tags (ESTs) were examined by various measures to assess the quality of these libraries. All ESTs were processed to remove cloning system sequences and contaminants and then assembled using CAP3. Following these processing steps, this assembly yielded 101,107 sequences derived from 89,043 clones, which defined 16,740 contigs and 33,213 singletons, a total of 49,953 "unigenes." Analysis of the distribution of these unigenes among the libraries led to the conclusion that the enrichment methods were effective in reducing the most abundant unigenes and to the observation that the most diverse libraries were from tissues exposed to environmental stresses including heat, drought, salinity, or low temperature.

The focus of this study was to analyze the content, distribution, and comparative genome relationships of 996 chromosome bin-mapped expressed sequence tags (ESTs) accounting for 2266 restriction fragments (loci) on the homoeologous group 3 chromosomes of hexaploid wheat (Triticum aestivum L.). Of these loci, 634, 884, and 748 were mapped on chromosomes 3A, 3B, and 3D, respectively. The individual chromosome bin maps revealed bins with a high density of mapped ESTs in the distal region and bins of low density in the proximal region of the chromosome arms, with the exception of 3DS and 3DL. These distributions were more localized on the higher-resolution group 3 consensus map with intermediate regions of high-mapped-EST density on both chromosome arms. Gene ontology (GO) classification of mapped ESTs was not significantly different for homoeologous group 3 chromosomes compared to the other groups. A combined analysis of the individual bin maps using 537 of the mapped ESTs revealed rearrangements between the group 3 chromosomes. Approximately 232 (44%) of the consensus mapped ESTs matched sequences on rice chromosome 1 and revealed large- and small-scale differences in gene order. Of the group 3 mapped EST unigenes approximately 21 and 32% matched the Arabidopsis coding regions and proteins, respectively, but no chromosome-level gene order conservation was detected.

We constructed high-density deletion bin maps of wheat chromosomes 5A, 5B, and 5D, including 2338 loci mapped with 1052 EST probes and 217 previously mapped loci (total 2555 loci). This information was combined to construct a consensus chromosome bin map of group 5 including 24 bins. A relatively higher number of loci were mapped on chromosome 5B (38%) compared to 5A (34%) and 5D (28%). Differences in the levels of polymorphism among the three chromosomes were partially responsible for these differences. A higher number of duplicated loci was found on chromosome 5B (42%). Three times more loci were mapped on the long arms than on the short arms, and a significantly higher number of probes, loci, and duplicated loci were mapped on the distal halves than on the proximal halves of the chromosome arms. Good overall colinearity was observed among the three homoeologous group 5 chromosomes, except for the previously known 5AL/4AL translocation and a putative small pericentric inversion in chromosome 5A. Statistically significant colinearity was observed between low-copy-number ESTs from wheat homoeologous group 5 and rice chromosomes 12 (88 ESTs), 9 (72 ESTs), and 3 (84 ESTs).

A total of 944 expressed sequence tags (ESTs) generated 2212 EST loci mapped to homoeologous group 1 chromosomes in hexaploid wheat (Triticum aestivum L.). EST deletion maps and the consensus map of group 1 chromosomes were constructed to show EST distribution. EST loci were unevenly distributed among chromosomes 1A, 1B, and 1D with 660, 826, and 726, respectively. The number of EST loci was greater on the long arms than on the short arms for all three chromosomes. The distribution of ESTs along chromosome arms was nonrandom with EST clusters occurring in the distal regions of short arms and middle regions of long arms. Duplications of group 1 ESTs in other homoeologous groups occurred at a rate of 35.5%. Seventy-five percent of wheat chromosome 1 ESTs had significant matches with rice sequences (E < or = e(-10)), where large regions of conservation occurred between wheat consensus chromosome 1 and rice chromosome 5 and between the proximal portion of the long arm of wheat consensus chromosome 1 and rice chromosome 10. Only 9.5% of group 1 ESTs showed significant matches to Arabidopsis genome sequences. The results presented are useful for gene mapping and evolutionary and comparative genomics of grasses.

A total of 1918 loci, detected by the hybridization of 938 expressed sequence tag unigenes (ESTs) from 26 Triticeae cDNA libraries, were mapped to wheat (Triticum aestivum L.) homoeologous group 4 chromosomes using a set of deletion, ditelosomic, and nulli-tetrasomic lines. The 1918 EST loci were not distributed uniformly among the three group 4 chromosomes; 41, 28, and 31% mapped to chromosomes 4A, 4B, and 4D, respectively. This pattern is in contrast to the cumulative results of EST mapping in all homoeologous groups, as reported elsewhere, that found the highest proportion of loci mapped to the B genome. Sixty-five percent of these 1918 loci mapped to the long arms of homoeologous group 4 chromosomes, while 35% mapped to the short arms. The distal regions of chromosome arms showed higher numbers of loci than the proximal regions, with the exception of 4DL. This study confirmed the complex structure of chromosome 4A that contains two reciprocal translocations and two inversions, previously identified. An additional inversion in the centromeric region of 4A was revealed. A consensus map for homoeologous group 4 was developed from 119 ESTs unique to group 4. Forty-nine percent of these ESTs were found to be homoeologous to sequences on rice chromosome 3, 12% had matches with sequences on other rice chromosomes, and 39% had no matches with rice sequences at all. Limited homology (only 26 of the 119 consensus ESTs) was found between wheat ESTs on homoeologous group 4 and the Arabidopsis genome. Forty-two percent of the homoeologous group 4 ESTs could be classified into functional categories on the basis of blastX searches against all protein databases.

The complex hexaploid wheat genome offers many challenges for genomics research. Expressed sequence tags facilitate the analysis of gene-coding regions and provide a rich source of molecular markers for mapping and comparison with model organisms. The objectives of this study were to construct a high-density EST chromosome bin map of wheat homoeologous group 2 chromosomes to determine the distribution of ESTs, construct a consensus map of group 2 ESTs, investigate synteny, examine patterns of duplication, and assess the colinearity with rice of ESTs assigned to the group 2 consensus bin map. A total of 2600 loci generated from 1110 ESTs were mapped to group 2 chromosomes by Southern hybridization onto wheat aneuploid chromosome and deletion stocks. A consensus map was constructed of 552 ESTs mapping to more than one group 2 chromosome. Regions of high gene density in distal bins and low gene density in proximal bins were found. Two interstitial gene-rich islands flanked by relatively gene-poor regions on both the short and long arms and having good synteny with rice were discovered. The map locations of two ESTs indicated the possible presence of a small pericentric inversion on chromosome 2B. Wheat chromosome group 2 was shown to share syntenous blocks with rice chromosomes 4 and 7.

Background: The combination of chemotherapy and hyperthermia (HT) is a promising approach in the treatment of malignant tumors. In the present report we evaluate the efficacy and toxicity of a combination of weekly paclitaxel combined with local hyperthermia in breast cancer., Patients and Methods: 7 patients were treated for inoperable local recurrence of breast cancer after mastectomy, irradiation, and chemotherapy or hormonal therapy. They weekly received paclitaxel (60-80 mg/m(2)) in 3-h infusions followed by local HT 41-44 degrees C for 45 min for 6-18 cycles., Results: Objective local response was observed in all treated patients (complete response in 4 patients and partial response in 3 patients). There were no grade 3 or 4 toxicities, neurologic toxicity or hypersensitivity reactions. Local tolerance to this regimen was also good, with only 4 patients developing mild transient erythema., Conclusion: Our experience indicates that the combination of weekly paclitaxel and HT may be effective in the treatment of locally recurrent breast cancer after mastectomy.

Aza-peptide Michael acceptors are a new class of irreversible inhibitors that are highly potent and specific for clan CD cysteine proteases. The aza-Asp derivatives were specific for caspases, while aza-Asn derivatives were effective legumain inhibitors. Aza-Lys and aza-Orn derivatives were potent inhibitors of gingipain K and clostripain. Aza-peptide Michael acceptors showed no cross reactivity toward papain, cathepsin B, and calpain.

Aims: The paper draws attention to the importance of rare, but important anatomical variety of nervus laryngeus recurrens, s.c. nervus laryngeus non-recurrens, for making thyroidectomy safely. Possible differences in the course of the nerve and the way of preparing the gland for minimizing the peroperation damage are presented., Patients: Three cases out of 1,200 operations on thyroid gland (0.25%) had the anatomical deviation described., Results: The nerve was saved from peroperation damage in two patients with nervus laryngeus recurrens. The third patient with this deviation ended up with permanent unilateral paralysis of the nerve, which indicated erroneous evaluation of nerve branches., Conclusions: Possible occurrence of rare varieties of nervus laryngeus recurrens point out necessary routine search and careful dissection during thyroidectomy. The diagnosis of nervus laryngeus non-recurrens is not possible in most cases. This contribution draws attention to basic operation procedures, which protect not only nervus laryngeus recurrens in its typical position, but also its varieties such as nervus laryngeus non-recurrens.

The main problem of the conventional radiotherapy in postoperative or curative irradiation in thyroid cancer is to achieve a sufficient dose in the planning target volume with acceptable dose distribution homogeneity and at the same time not to exceed tolerance doses in the organs of risk. Our study presents the advantages of intensity-modulated radiotherapy over conventional radiotherapy on an example of dosimetric comparisons of isodose plans of three patients treated at our department. The intensity-modulated radiotherapy in thyroid cancer offers an improvement of dose homogeneity and better sparing of organs at risk (spinal cord and lungs). Furthermore, dose escalation is feasible in the whole initial planning target volume up to 60 Gy with a simultaneous sparing of healthy tissues and organs at risk.

Background: Serum copper concentrations as indicators of non-specific inflammatory response are higher in patients with severe heart failure than in patients with mild to moderate cardiac insufficiency and correlate with acute phase protein concentrations. The aim of this study was to find out if baseline serum copper concentrations are higher in patients with chronic heart failure and higher one year mortality and morbidity and whether serum copper concentrations correlate with prognostic indicators., Methods and Results: Baseline serum copper concentrations were determined in 64 patients with chronic heart failure, functional classification NYHA II-IV (NYHA - New York Heart Association), caused by ischemic heart disease, with severe left ventricular dysfunction (ejection fraction < or = 35%). 30 patients died or were admitted to the hospital because of worsening heart failure (group A) within 12 months since their examination. 34 patients did not die and hospitalisation was not necessary within 12 months (group B). Retrospectively the presence of differences in serum copper concentrations and in prognostic indicators was assessed between both groups. Statistically significant differences between both groups were found in these parameters: baseline serum copper concentrations (p < 0.001), ejection fraction of left ventricle (p < 0.05), baseline heart rate (p < 0.01) and cardiothoracic ratio (p < 0.01). Serum copper concentrations significantly correlate with heart rate (p < 0.01), inverse correlation with left ventricular ejection fraction and positive correlation with cardiothoracic ratio are not statistically significant., Conclusions: Baseline serum copper concentrations are significantly higher in patients with chronic heart failure, also correlating with higher one-year mortality and morbidity. Serum copper levels significantly correlate with baseline heart rate. Relation of serum copper concentrations with left ventricular ejection fraction and cardiothoracic ratio is not statistically significant.

The retrospective study was aimed at demonstrating anatomical and functional results of surgical treatment in patients with bilateral idiopathic retinal dialysis (BIRD). In the period of 1985-2001 the authors operated on BIRD in 10 eyes of five patients (three men and two women). No eye injury or inflammation was present in the case history of the patients. The age of the patients was between 11 and 30 years, the mean age was 19.2 years. The symptoms typical for retinal detachment were present in four eyes only and absent in the other six eyes. Hyperopia of both eyes in analogy with myopia of both eyes was observed in two patients, respectively, whereas emmetropia was present in one patient. The retinal dialysis (RD) occurred in lower temporal quadrant in all eyes (100%), in one eye (10%) it also occurred together with an discontinuous RD in upper temporal quadrant and in another eye it was extended into lower nasal quadrant. Demarcation lines were present in 5 eyes (50%), subretinal solid strands in 1 eye (10%). Scleral buckling procedures were performed as the primary surgical procedure in all 10 eyes of 5 patients. Eight eyes was operated on with the use of segmental circumferential sponge buckles of silicone and two eyes with the use of encircling elements of solid silicone. Retina was successfully reattached postoperatively in 9 eyes (90%) and remained detached in one eye (10%). The visual acuity preoperatively 6/12 or better remained unchanged postoperatively in 8 eyes (80%). The visual acuity postoperatively improved in 1 eye (10%) and in 1 eye (10%) visual acuity was decreased. The mean period of observation of the patients was 86 months. The necessity of a routine examination of the extreme retinal periphery of the fellow asymptomatic eye of the young adult with no history of trauma and with the retinal detachment associated with retinal dialysis in one eye is also discussed.

A novel reversed-phase high-performance liquid chromatographic method with UV spectrophotometric detection was developed and validated for the determination of compounds in topical cream. The method describes determination of active component hydrocortisone acetate (HCA), its degradation products hydrocortisone (HC) and cortisone acetate (occurring in formulation after long-term stability tests) and two preservatives presented in the cream-methylparaben and propylparaben, using dexamethasone as an internal standard. The chromatographic separation was performed on a 5 microm SUPELCO Discovery C18 125 x 4-mm ID column. The optimised mobile phase for separation of all the compounds consists of methanol, acetonitrile and water (15:27:58, v/v/v), with the analysis time less than 13 min. The method was applicable for routine analysis (assays and stability tests) of active compound HCA, preservatives and degradation products in pharmaceutical product--topical cream Hydrocortizone cream 1%.

The method of choice in the treatment of penetrating eye injuries affecting the vitreous body with an intraocular foreign body is in the majority of patients primary pars plana vitrectomy with transvitreal extraction of the foreign body (FB). During the period from April 1999 till September 2000 the authors treated thus at the Ophthalmological Clinic of the General Faculty Hospital and 1st Medical Faculty Charles University Prague 26 eyes of 26 young men with an average age of 34 years. The mean interval between the injury and primary PPV was 8.5 days. The patients were followed up on average for 23.5 months. The entrance of the penetrating injury was in 17 eyes in the cornea or limbus, in 9 eyes in the sclera. In two eyes posttraumatic endophthalmitis developed. The intraocular FB was 6 mm2 in size in 16 eyes and 6-20 mm2 in 10 eyes. In primary PPV we indicated in 14 eyes internal tamponade with gas or silicone oil. On account of late retinal complications which developed as a result of proliferative vitreoretinopathy we indicated in eight eyes PPV reoperation. At the end of the investigation period in 3 eyes inoperable detachment of the retina developed. As the main negative prognostic factor for final visual acuity the authors evaluated a size of the FB greater than 6 mm2, other negative prognostic factors are the scleral entrance injury, complications of the entrance injury (prolapse of the iris, vitreous body, haemophthalmus) and the development of posttraumatic exogenous endophthalmitis.

Unlabelled: Acute exogenous endophthalmitis (AEE) is the most serious post-traumatic or postoperative complication which without immediate treatment leads to amaurosis or anatomical loss of the eye. If comprehensive antibiotic treatment fails immediate pars plana vitrectomy (PPV) is indicated. Empirically and later also by laboratory methods the antiinflammatory effect of silicone oil (SO) was revealed. The objective of the present retrospective study is to demonstrate anatomical and functional results of PPV with implantation of SO for AEE on eyes without detachment of the retina. In 1990-2000 the authors operated a total of 22 eyes of 22 patients with AEE. The mean age of the patients was 59.6 years, the mean observation period 35.5 months. In 17 eyes postoperative and in 5 eyes post-traumatic AEE was involved. The mean interval between the original operation and PPV was 20.8 days and between the injury and PPV 5.4 days. In all patients we took at the beginning of PPV specimens of the vitreous body for cultivatin which gave in 63.6% a positive result. The intraocular lens was explanted in 81.8% eyes. After subtotal resection of the vitreous body we implanted SO and in all aphakic eyes we performed iridectomy in no. 6. PPV was completed by administration of vancomycin and ceftazidime into the vitreous space. In 95.4% of the eyes we preserved anatomically the bulbus, in one instance we eviscerated the bulbus three days after failure of PPV (4.6%). In 86.4% the retina remained anatomically attached and in the remaining 9% eyes beneath the SO traction detachment of the retina developed. Visual acuity improved in 72.7% eyes, it remained unchanged in 9.1% eyes and deteriorated in 18.2% eyes. Silicone oil was explanted in 40.9% eyes after a mean interval of 4.6 months following PPV. In the remaining patients we did not indicate explantation of SO, in particular because of the patients' advanced age,, Conclusion: We recommend PPV and implantation of SO in AEE on eyes without detachment of the retina as the method of choice.

Vascular restenoses currently represent a major problem in the treatment of vascular stenoses. One of new approaches in the prevention of restenoses is intravascular brachytherapy. Intravascular brachytherapy uses local irradiation of the stenotic vessel segment by ionizing radiation with the aim of prevention of restenosis. This is a new rapidly developing multidisciplinary approach based on collaboration of specialties of intervention radiology, intervention cardiology, angiology, nephrology and radiation oncology. This review examines current options of intravascular brachytherapy as well as results of clinical trials evaluating the efficacy of intravascular brachytherapy in the different anatomical regions. Intravascular brachytherapy may substantially reduce the rate of restenoses. However, intravascular brachytherapy should be currently used only in the setting of clinical trials. Optimal method of irradiation of the stenotic segment of the vessel is still to be defined.

The insulin-like growth factor 2 gene (IGF2) has been described in several studies as a candidate gene for meat efficiency in pigs. IGF2 is a member of the growth factors family and has an effect on development of muscle tissue. The effect of IGF2 gene polymorphism on meat efficiency was analysed in a population of 121 Large White pigs. A PCR-based test and RFLP methods were used for detection of genotypes. Allele A, lacking the restriction site, was characterised by the presence of a 0.9-kb fragment. In allele B, the amplimer was cut into a 0.8-kb fragment and some barely detectable fragments. The statistical analysis was carried out according to the General Linear Model procedure. The genotype frequencies observed were: 1.65%, 33.88%, 64.46% for AA, AB and BB genotypes, respectively. There was a significant difference (P < or = 0.05) between the AB and BB genotypes in live weight before the test. A significant association between AB and BB genotypes and body weight before the test was found. No significant difference in other traits of growth and meat efficiency was observed (P > 0.05).

Background: Liver is the most common site of metastatic disease. Hepatic arterial infusion (HAI) of cytotoxic drugs may achieve high objective response rate., Methods: Peripheral blood mononuclear cells were obtained by leukapheresis after stimulation with subcutaneous GM-CSF in six patients and with subcutaneous interleukin-2 (IL-2) in four patients, all with nonresectable hepatic metastases not responsive to conventional regimens. After the cytokine stimulation, the cells were administered by HAI either alone, or after HAI of melphalan (50 mg)., Results: Mean number of 23.1 +/- 4.6 x 10(9) and 19.0 +/- 9.7 x 10(9) mononuclear cells were obtained through leukapheresis after GM-CSF and IL-2 priming, respectively. Significant cytotoxic activity was observed only after short IL-2 stimulation. A marked decrease in tumor markers was observed in two patients treated by combination of melphalan and activated leukocytes., Conclusions: HAI is a technically feasible way of regional delivery of a high number of activated leukocytes obtained after short ex vivo activation.

Bone metastasis is an unusual complication of hepatocellular carcinoma. We report here 2 cases of patients with bone metastases of hepatocellular carcinoma at presentation. Patient No. 1 with liver cirrhosis and hepatocellular carcinoma was admitted with a bone metastasis in the rib. The patient was treated with hepatic arterial chemotherapy and rib resection. Patient No.2 was known to have an asymptomatic liver mass of uncertain histology for a year when he presented with back pain. Because of signs of spinal compression, laminectomy was performed, and the diagnosis of metastatic hepatocellular carcinoma was established. The presence of bone metastases in hepatocellular carcinoma at presentation is extremely rare. More frequently, bone lesions are observed after successful treatment of the primary liver tumor. Both surgery and radiotherapy are used as palliative treatment in bone metastases of hepatocellular carcinoma. The treatment of hepatocellular carcinoma presenting with bone metastasis by bone resection and intraarterial chemotherapy seems to be of limited impact on patient survival because of dissemination of micrometastases in other organs and the frequent presence of other comorbid conditions. However, effective palliation using this multimodality approach is feasible. Hepatocellular carcinoma should be considered in the differential diagnosis of bone metastases.

Background/aims: The prognosis of biliary tract carcinomas is poor, and therapeutic efforts are limited mostly only to palliation. The aim of this study was to retrospectively evaluate the effectiveness and tolerability of intraluminal high dose rate brachytherapy in the treatment of bile duct and gallbladder carcinomas., Methodology: Thirteen patients with bile duct and gallbladder carcinomas were treated by brachytherapy administered through high dose rate remote afterloading system. Five patients after Roux-en-Y hepaticojejunoanastomosis were treated by intraluminal brachytherapy inserted via a diahepatal drain, and 8 inoperable patients were treated by intraluminal brachytherapy via a percutaneous biliary drain., Results: After intraluminal brachytherapy, a control of icterus was observed in all patients. The treatment was well tolerated and mean survival was 275 days., Conclusions: The addition of intraluminal brachytherapy may be beneficial to patients with carcinomas causing biliary obstruction in whom bile drainage can be established.

Background/aims: The prognosis of biliary tract cancers is poor. The aim of the present report was to analyze retrospectively the effectiveness of regional chemotherapy in patients with biliary tract cancers treated at a single institution., Methodology: Thirty-two patients with biliary tract cancers, 17 patients with cholangiocarcinoma and 15 patients with gallbladder carcinoma, were treated by regional administration of the chemotherapy, usually the combination of 5-fluorouracil, cisplatin and folinic acid. Eighteen patients with inoperable tumors received no surgical treatment, 10 patients were treated by palliative resection (cohort B), and 4 patients received radical surgery (cohort C)., Results: The median survival of the patients is (mean +/- standard deviation) 14 +/- 17+ (median 7+) months for cohort A patients, 22 +/- 17+ (median 17+) months for cohort B patients, and 32 +/- 4+ (median 33+) months for cohort C patients. One-year survival was 38% (6 out of 16 patients) for cohort A, 80% (8 out of 10 patients) for cohort B, and 100% (4 out of 4 patients for cohort C). Two-year survival was 15% (2 out of 13) for cohort A, 30% (3 out of 10) for cohort B, and 100% (4 out of 4) for cohort C. One out of 12 patients (8%) in cohort A and 1 out of 10 patients (10%) in cohort B survived more than 5 years. After intraarterial chemotherapy, a significant increase was observed in the absolute numbers of CD3+, CD3+CD8+ and CD8+CD28+ lymphocytes, as well as an increase in CD3+CD4+ and natural killer lymphocytes., Conclusions: Regional chemotherapy is active in controlling the disease, and seems to result in prolongation of survival in patients with biliary tract cancer. Administration of regional chemotherapy is also associated with a rise of circulating lymphocyte numbers.

Background/aims: The prognosis of carcinomas of the subhepatic region is poor, and therapeutic efforts are limited mostly to palliation. The aim of this study was to retrospectively evaluate the effectiveness of transduodenal administration of intraluminal high dose rate brachytherapy in the palliative treatment of bile duct and pancreatic head carcinomas., Methodology: Twelve patients with inoperable bile duct and pancreatic head carcinomas were treated by transduodenal brachytherapy using high dose rate remote afterloading system. Eleven patients were treated by intraluminal brachytherapy inserted via a nasobiliary drain and one patient by intraluminal brachytherapy via a nasopancreatic drain inserted in the duct of Wirsung., Results: After transduodenal intraluminal brachytherapy, a control of icterus was observed in all patients. The treatment was well tolerated with the mean survival of 284 days., Conclusions: Transduodenal intraluminal brachytherapy is technically feasible. The addition of intraluminal brachytherapy may be beneficial to patients in whom drainage can be established. Transduodenal insertion of brachytherapy is not competitive to the percutaneous approach but spreads the possibilities of the treatment of bile duct carcinoma. Intraluminal brachytherapy of pancreatic head carcinoma is feasible only via transduodenal approach.

Objective: Based on their on experience the authors express their opinion on routine detection and visualization of the NLR during radical surgery of the thyroid gland. They describe standard and alternative steps of surgery leading to protection of the recurrent nerve., Material and Methods: The authors evaluated a group of 735 patients operated during the last thirty nine months which comprised 87% total thyroidectomies, 11.6% total lobectomies, the remainder were almost total operations. The number of visualized nerves was 1367., Results: Unilateral temporary damage of the NLR leading to temporary pareses of the vocal cords was recorded in 19 patients, i.e. 1.39%, bilateral transient paresis in one case, i.e. 0.07%. Permanent damage of one vocal cord after severing of the NLR occurred in 11 patients, i.e. 0.8%, bilateral lesions of the vocal cords were not recorded., Conclusion: There are no extensive statistics of thyroid operations without reports on damage of the NLR. With the increasing number of radical operations of the gland also the number of departments increased which recommend unequivocally to seek and visualize the NLR. The authors results support this point of view. They use most frequently a modification of Wang's method and consider as the smallest extent of visualization of the NLR the distance between the branching of the inferior thyroid artery and the entry of the NLR into the larynx.

A 76-year-old man was admitted to the surgical department with acute abdominal pain and impaired sensation of the lower extremities. An aneurysm of the abdominal aorta (AAA) was diagnosed already in the past. There were no signs of cardiovascular failure. Examination (sonography, CT) did not show intraabdominal bleeding. Nevertheless AAA rupture was suspected. A decision on an urgent operation was taken. Despite permanent resuscitation the patient died on the table before the operation began. In the discussion four type of AAA rupture are mentioned: into the open abdominal cavity, into the retroperitoneum, into surrounding organs such as gut or vena cava and so-called "sealed rupture". In every symptomatic AAA connected with circulatory instability rupture is suspected. An urgent operation is necessary in these cases. Rupture of AAA into the vena cava inferior is rare. A syndrome including a history of aneurysm, abdominal pain, continuous abdominal murmur and heart failure is pathognomic for this type of rupture. Computer tomography, sonography or arteriography could be helpful the diagnose determination. However, correct preoperative diagnosis is difficult. Other causes of circulatory failure, especially heart attack, must be differentiated. Treatment of such cases is surgical, using a stent graft is rare and determined only for indicated cases.

The dependence of the impedance of the electrode double layer of mercury electrode on frequency around the potentials of the tensammetric peaks of single-stranded and double-helical polynucleotides and DNA was studied. From the frequency dependence of the impedance of the electrode double layer represented in a complex plane impedance plot, the electric equivalent circuit of the electrode covered with adsorbed DNA layer was determined. It was concluded that the desorption of denatured ssDNA is accompanied by higher dielectric losses than the desorption of native dsDNA. This can be explained by the higher flexibility of ssDNA compared to the dsDNA. The capacitance peak of single-stranded polyadenylic acid (poly A) observed at pH 8 around -1.3 V splits at low frequencies in two peaks.

Unlabelled: The authors quantified the changes of left ventricular kinetics during hospitalization after a first transmural myocardial infarction treated by direct percutaneous coronary angioplasty. For accurate quantification a group as close as possible to experimental conditions was selected. 64 patients (43 men), mean age 67.3 years (44-86) were investigated. The echocardiographic examination was made within 24 hours after admission and on discharge (9.8 days). A 16-segment of the left ventricle was used, the kinetics were evaluated by 4-grade score. The ejection fraction was calculated by Simpson's method from 4 cycles, the wall motion score index was calculated by the standard procedure. During the follow up period significant improvement of left ventricular kinetics occurred. The ejection fraction improved from the original 46 +/- 11% (median 41.5) to 51 +/- 11% (median 50.0) (p < 0.005), the wall motion score index from 1.6 +/- 0.3 (median 1.65) improved to 1.5 +/- 0.3 (median 1.60) (p < 0.005). Improvement of the investigated parameters correlated with the clinical course, 87% of the patients being free of any complications., Conclusion: Successful direct angioplasty leads in patients with a first myocardial infarction to improvement of the ejection fraction and wall motion score index already during the first 10 days.

The capacitance measurement (dependence of the differential capacitance C of the electrode double layer on potential E, C-E curves), electrochemical impedance spectroscopy (frequency response of the impedance Z of the electrode double layer-EIS) and constant current chronopotentiometry (dependence of dt/dE on potential at constant current, chronopotentiometric stripping analysis-CPSA) have been used for electrochemical study of echinomycin and its interaction with single-stranded (ss) and double-stranded (ds) DNA at the hanging mercury drop electrode (HMDE). The capacitance measurement showed that echinomycin gives a pseudocapacitance redox peak strongly dependent on the a.c. voltage frequency at the potential of -0.53 V. This peak is observed with dsDNA-echinomycin complex as well, but not with ssDNA treated by echinomycin. Similar results were obtained using CPSA measurements. Thus capacitance measurements and CPSA can distinguish with the aid of the bis-intercalator echinomycin the single-stranded and double helical form of DNA adsorbed at the mercury electrode surface. Impedance measurement in connection with adsorptive transfer technique can find the differences between ssDNA and dsDNA, which promise to use this technique for detection of dsDNA in hybridisation reactions.

Schistosomes are parasites of considerable medical and veterinary importance and, therefore, all aspects of their biology have been intensively studied. In contrast, our knowledge of species of the largest genus, Trichobilharzia, is insufficient. Because morphological characterization of Trichobilharzia species provides a limited number of criteria for species determination, molecular data are required. In the present paper, we sequenced internal transcribed spacers ITS1 and ITS2, and 5.8S ribosomal RNA (rRNA) genes of 3 European Trichobilharzia species (T. regenti, T. szidati and T. franki). We showed that ITS1 and ITS2 sequences can be used in species identification. Repetitive elements were found in ITS1 of all 3 Trichobilharzia species; their number and length varied depending on the species. Phylogenetic analysis showed that the visceral T. franki is more related to the nasal T. regenti, than to the visceral T. szidati. The newly designed primer, which is specific for T. regenti, might be used as a tool for diagnosis of this potential pathogen.

Aims: Direct angioplasty is an effective treatment for ST-elevation myocardial infarction. The role of very early angioplasty in non-ST-elevation infarction is not known. Thus, a randomized study of first day angiography/angioplasty vs early conservative therapy of evolving myocardial infarction without persistent ST-elevation was conducted., Methods: One hundred and thirty-one patients with confirmed acute myocardial infarction without ST-segment elevations were randomized within 24 h of last rest chest pain: 64 in the first day angiography/angioplasty group and 67 in the early conservative group (coronary angiography only after recurrent or stress induced myocardial ischaemia)., Results: All patients in the invasive group underwent coronary angiography on the day of admission (mean randomization-angiography time 6.2 h). First day angioplasty of the infarct related artery was performed in 47% of the patients and bypass surgery in 35%. In the conservative group, 55% underwent coronary angiography, 10% angioplasty and 30% bypass surgery within 6 months. The primary end-point (death/reinfarction) at 6 months occurred in 6.2% vs 22.3% (P<0.001). Six month mortality in the first day angiography/angioplasty group was 3.1% vs 13.4% in the conservative group (P<0.03). Non-fatal reinfarction occurred in 3.1% vs. 14.9% (P<0.02)., Conclusions: First day coronary angiography followed by angioplasty whenever possible reduces mortality and reinfarction in evolving myocardial infarction without persistent ST-elevation, in comparison with an early conservative treatment strategy., (Copyright 2001 The European Society of Cardiology.)

Background: Currently, there is no standard treatment of inoperable advanced hepatocellular carcinoma., Study: A patient with advanced hepatocellular carcinoma was treated with intravenous infusion of pegylated liposomal doxorubicin (PLD, Caelyx) in combination with ultrasound hyperthermia of the liver. Each cycle consisted of infusion of 60 mg of PLD followed by two fractions of hyperthermia 41 degrees C to 43 degrees C for 45 minutes 1 and 48 hours after infusion, respectively., Results: A substantial regression of the tumor was observed on computed tomography scans. No toxicity of combined treatment was noted., Conclusions: This may be the first report of the combination of PLD and hyperthermia in the treatment of advanced hepatocellular carcinoma. Our observation suggests that the combination of PLD with hyperthermia is technically feasible, well tolerated, and could have synergistic potential.

Liver arterial system in humans presents wide variability. Knowledge of the variability is important for operation in this region (e.g. port-catheter implantation for regional intraarterial chemotherapy in patients with liver malignancy). The purpose of this study is to determine types of variation of liver artery system and their frequency in Czech population. The research was based on the evaluation of abdominal arteriograms, arteriography findings were confirmed during operations. There were found various anatomical relationships in this area, which may be divided into five types: type I.--standard organization occurred in 75.6% of cases; type II.--presence of accessory artery (8.2%); type III.--variation of hepatic artery origin (7.4%); type IV.--the anomalous origin of gastroduodenal artery (6.9%) and type V.--combination of varieties (1.9%). The most often was the type I that occurred in three quarters of cases approximately.

Background/aims: The prognosis of malignant melanoma metastatic to the liver is poor. The aim of the present report was to analyze retrospectively the effectiveness of regional chemotherapy and biologic therapy in patients with hepatic metastases of malignant melanoma., Methodology: Seven patients with hepatic metastases of malignant melanoma were treated by intraarterial administration of the combination of cisplatin, vinblastine and dacarbazine, or melphalan, with or without interleukin-2, interferon-alpha and interferon-gamma., Results: The median survival of 4 patients with metastatic involvement initially limited to the liver is 19+ months in contrast to a median survival of 5 months in patients with concurrent extrahepatic disease. Intraarterial administration of cytokines led to an initial decrease in circulating lymphocyte numbers, with subsequent return to pretreatment levels, and to an increase in urinary neopterin, a marker of immune activation., Conclusions: Regional intraarterial administration of chemotherapy with or without cytokines may be effective for controlling hepatic metastases of malignant melanoma in patients with disease limited to the liver, but little benefit is evident in patients who present with concurrent extrahepatic disease.

Background/aims: Regional chemotherapy represents an effective approach for the control of isolated liver metastases of colorectal cancer. Fluoropyrimidines have been the basis of systemic and regional chemotherapy of this disease for several decades, but recent studies have demonstrated that the addition of irinotecan (CPT-11) ameliorates the results of systemic treatment., Methodology: Fifteen patients with isolated liver metastases of colorectal carcinoma were treated with regional administration of CPT-11 in combination with 5-fluorouracil/folinic acid (5-FU/FA). CPT-11 (100-200 mg, mean: 93 +/- 25, range: 51-125 mg/m2) was administered weekly in combination with 5-FU (1000-2000 mg, 857 +/- 182, range: 538-1301 mg/m2) and FA (100-350 mg). Peripheral blood leukocyte phenotype was examined during the treatment in selected patients., Results: One patient achieved complete response, 4 patients had partial response, 7 patients had stable disease, 1 patient progressed and 2 patients were not evaluable. The response rate was 38% in evaluable patients, and a more than 50% decrease in serum carcinoembryonic antigen levels was observed in 12 out of 14 patients. The treatment was generally well tolerated. All patients, except one, are currently alive at mean follow-up of 11 +/- 6 (median: 10, range: 5-24) months. The therapy is still being continued in 8 patients., Conclusions: Weekly CPT-11/5-FU/FA is highly effective in the treatment of patients with metastatic colorectal cancer limited to the liver, even after failure of previous 5-FU/FA. The present regimen should be tested as a first line treatment in phase III trial.

Background: The combination of hyperthermia and radiotherapy has additive or synergistic effects. This combination has been studied extensively in radiation oncology, but not in the prevention of vascular restenosis., Case Report: A patient with restenosis of cephalic vein underwent percutaneous transluminal angioplasty (PTA) followed by endovascular irradiation with 192Iridium (12 Gy) using a high dose rate afterloading technique. After endovascular irradiation, one fraction of external ultrasound hyperthermia was administered to the irradiated segment. There was no restenosis in the treated vessel segment according to duplex sonography performed 192 days after treatment. No radiation or hyperthermia associated side effects were observed., Conclusions: Present observations suggest that endovascular brachytherapy of restenosis potentiated by hyperthermia is a technically feasible and well-tolerated treatment. The additive and synergistic effects of hyperthermia, in conjunction with radiation, could be of benefit in the prevention of vascular restenosis.