Inter laboratory comparison for reliability and reproducibility of plant DNA flow cytometry.
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DNA, Plant↗
Biomedical subjects
J Dolezel
Publications and source records attributed to J Dolezel.
Construction of chromosome-specific DNA libraries covering the whole genome of field bean (Vicia faba L.).
Recombinant DNA libraries were constructed for seven chromosome types isolated from two translocation lines of field bean (Vicia faba L.) with reconstructed karyotypes. The chromosomes were selected so that the set of libraries covers the whole V. faba genome more than once. Individual chromosome types were highly purified by flow sorting, and their DNA was amplified by degenerate oligonucleotide-primed (DOP) polymerase chain reaction (PCR) and cloned into a plasmid vector. The choice of restriction site present in PCR primer and refinement of cloning protocol resulted in high cloning efficiency and allowed generation of libraries consisting of about 10(5) clones from 250 or 1000 sorted chromosomes. The insert size ranged between 50 and 2200 bp and the mean length estimated in individual libraries varied between 310 and 487 bp. Hybridization of cloned fragments with labelled genomic DNA showed that about 60% of inserts represented unique or low-copy sequences. The suitability of the libraries for genome mapping was demonstrated by isolation of clones containing microsatellite motifs.
Chromosome Mapping↗
[Ureteroscopy--an alternative treatment in ureterolithiasis].
The authors evaluate the results of 315 ureteroscopies performed in 1991-1995 at the Urological Clinic in Brno Bohunice. 290 ureteroscopies were made on account of ureterolithiasis, diagnostic ureteroscopy was performed in 17 patients, in four instances extraction of a foreign body was involved and in four instances during ureteroscopy the stenosis of the ureter was severed. One-stage operations were successful in 226 patients (78%), in 42 patients (14%) after ureteroscopy ESWL had to be made. Ureteroscopy as a two-stage operation was performed in 12 patients (6%), five cases were resolved by open surgery because ureteroscopy failed. The authors evaluate early and late complications, none of which called for nephrectomy. The authors discuss the position of ureteroscopy in ureterolithiasis which is one of the most frequent indications.
Adult↗
Sex determination in dioecious plants Melandrium album and M. rubrum using high-resolution flow cytometry.
High-resolution flow cytometry was used to analyze nuclear DNA content in dioecious plants Melandrium album and M. rubrum. Very low coefficients of variation (0.53-0.70%) of G0/1 peaks were obtained after the analysis of nuclei isolated by chopping leaf tissues. In both species, females have lower DNA content than males due to sex chromosome heteromorphism. This made it possible to discriminate peaks belonging to female and male nuclei on the same histogram and permitted unambiguous sex identification in both species. The results indicated the possibility of diagnosing sex also in other heterogametic dioecious plants. The assay described here might be interesting in any area where rapid sex identification is required at an early stage of plant development.
DNA, Plant↗
Cell cycle synchronization, chromosome isolation, and flow-sorting in plants.
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Cell Cycle↗
Primer-induced labeling of pea and field bean chromosomes in situ and in suspension.
A protocol for primed in situ DNA labeling (PRINS) was optimized for pea (Pisum sativum L.) and field bean (Vicia faba L.) chromosomes attached to coverslips. Cloned DNA or synthetic oligonucleotides were used as probes for repetitive DNA sequences (rDNA, Fok-element) and different reaction conditions were tested to achieve the highest specific signal-to-background ratio. A procedure based on direct labeling by fluorescein-dUTP was compared with an indirect one using digoxigenin detected by fluorescently labeled antibody. Under optimal conditions, strong and specific signals were obtained exclusively on chromosome regions known to contain respective DNA sequences. Compared to the direct labeling, significantly stronger signals were obtained when the indirect procedure was used. Both types of labeling were successfully applied to chromosomes in suspension and were shown to produce signals comparable to that obtained with chromosomes attached to coverslips. It is expected that primed in situ DNA labeling en suspension (PRINSES) will provide a basis for flow-cytometric discrimination and sorting of otherwise indistinguishable chromosomes according to their specific fluorescent labeling.
Base Sequence↗
[Implantation of inflatable penile prostheses as an effective alternative surgical treatment for erectile disorders].
The authors present an account of their own experience with results and complications of inflatable penile prostheses implanted in a group of 30 patients with organic erectile dysfunction. They discuss possible complications of this operation as well as the place of penile prostheses in the treatment of erectile dysfunction.
Adult↗
[An unusual case of plastic penile induration].
The presence of calcified plaques is one of the few unequivocal indications for surgery of Peyronie's disease. The mean size of plaques described in the literature is 1.5-2 cm whereby it may vary from 1 mm all along the penis. The authors treated successfully an unusually large calcified plaque 7 x 0.8 cm with excellent postoperative results--restoration of the patient's full sexual activity which was not possible before operation.
Humans↗
Localization of MPM-2 recognized phosphoproteins and tubulin during cell cycle progression in synchronized Vicia faba root meristem cells.
MPM-2 antibody reacts with a subset of mitotic phosphoproteins. We followed localization of MPM-2 immunoreactive material and localization of microtubules during cell cycle progression in a highly synchronous population of Vicia faba root meristem cells and isolated nuclei. The MPM-2 antibody labelling showed significant cell cycle dependence. MPM-2 nuclear reactivity was weak and homogeneous in G1 and S phase of the cell cycle and became stronger and heterogeneous during G2, resembling staining of the nuclear matrix, with maximum staining at the G2/M interface. Similarly the staining intensity of nucleoli increased from late G1 phase to nucleoli dispersion in early prophase. During mitosis MPM-2 immunoreactivity was associated with spindle configurations and the brightest signal was localized in kinetochores from prophase to metaphase.
Antibodies, Monoclonal↗
Localization of seed protein genes on flow-sorted field bean chromosomes.
Chromosomes from reconstructed field bean (Vicia faba L.) karyotypes were flow-sorted and the DNA was used for the physical localization of seed storage and nonstorage (USP) protein genes using PCR with sequence specific primers. The data were confirmed and refined by using DNA of microisolated chromosomes of other karyotypes as the target for PCR. The specificity of the PCR products was proved by restrictase digestion into fragments of predicted length or by reamplification using 'nested' primers. The genes are located within defined regions of chromosome I (USP = unknown seed protein genes), II (vicilin genes, legumin B3 genes), III (legumin B4 genes), IV (pseudogenes psi 1) and V (legumin A genes and pseudogenes psi 1). Except for the pseudogene derived from the sequence of legumin B4 gene, all members of each gene family are located in one chromosome region exclusively. This approach proved to be useful for localizing genes that cannot be mapped genetically (due to the lack of allelic variants) and might be applied to integrate physical and genetic maps.
Base Sequence↗
Very rare angioleiomyoma of the male urethra.
Herein is reported a case of angioleiomyoma, i.e. a benign tumour arising predominantly from smooth muscle cells of the vessel wall, in a yet unreported locality, the urethra. The closest to the tumour presented by us are leiomyomas of the urethra, so far described only in females. The angioleiomyoma of our case manifested itself as a stricture of the male urethra and resection operation showed to be the best approach with elimination of the patient's complaints.
Angiomyoma↗
Metastasis of Grawitz's tumour in the ureteral stump.
Metastasis of a tumour in the ureteral stump after nephrectomy for a primary tumour is very rare. Only 44 cases have been published so far. Our paper reports on a metastasis of Grawitz's tumour in the ureteral stump diagnosed 28 years after nephrectomy for a primary tumour. The authors have shown a low biological activity in this tumour.
Aged↗
Ureteral incarceration in the symptomatology of hydronephrosis.
Renal colic symptoms may be caused by an aberrant vascular bundle that intermittently incarcerates the pyeloureteral junction. The authors describe 6 patients who were complaint free except for the period of fits and routine urological examinations showed normal findings. The patients had no subjective complaints after open surgery.
Adult↗
[Ureteroscopy in the treatment of ureterolithiasis].
After establishment of endourological methods at our University Urological Department we have treated 36 ureteral stones in 34 patients since January 91 until April 92. Through retrograde (23) and antegrade (13) access we have successfully treated almost all calculi with the exception of one (where we hadn't been able to pass through reimplanted ureteral orificium). We have introduced double J stent in 90% patients after procedure and discharged them 3 days postoperatively on average. We have had no serious complication. Ureteroscopy with stone disintegration possibility has enabled us almost to leave open surgery in those cases, lower morbidity and shorten time spent in the hospital significantly.
Endoscopy↗
Staining of fungal cell walls with fluorescent brighteners: flow-cytometric analysis.
Spore walls of Backusella lamprospora (Mucorales) were stained with ten fluorescent brighteners (FB) and the intensity of their fluorescence was determined. The fluorescence was most intense with Uvitex 2B (100%), other brighteners yielding lower fluorescence intensities: Blankophor BA 267% and BA 200% about 75%, Rylux BSU about 50%, other Rylux agents 10-30%. The agents most suitable for microscopic diagnostics of human and animal mycoses are Uvitex 2B, Blankophor BA 267% and BA 200%, Rylux BSU, and also Rylux BS and PRS. The regulation of excessive fluorescence of fungal cells during microscopic observation is discussed. For the purposes of microscopic diagnosis of human and animal mycosis Uvitex 2B, Blankophor BA 267% and BA 200%, Rylux BSU and, possibly, Rylux BS and PRS are recommended.
Animals↗
Improvement of accuracy of scanning absorption measurement of nuclear DNA content in plants.
The effect of optical errors and stain variation on the accuracy of plant Feulgen-DNA absorption cytophotometry was investigated. Optical errors (glare, distributional error, and diffraction) were shown to cause important errors. A correction procedure of DUIJNDAM et al. (1980a) was successfully applied to eliminate them. Stain variation can also lead to serious errors and in case different samples are to be compared the use of an internal standard is imperative. Chicken red blood cells proved to be a reliable standard.
Cell Nucleus↗
Sequential estimation of nuclear DNA and silver staining of nucleoli in plant cells.
A method for sequential estimation of nuclear DNA and silver staining of nucleoli in plant cells is described. Feulgen staining is done first and nuclear DNA estimated by absorption cytophotometry. Following this, the slides are stained with AgNO3. The method has been used to study the process of nucleolar fusion in garlic (Allium sativum L.) meristem root tip cells. It was found that during interphase nucleoli rarely fused, thus most fusion must have occurred before the G1 phase of the cell cycle.
Coloring Agents↗
Importance of blood clotting tests in the diagnosis of haemolytic-uremic syndrome and experience with its treatment.
More than half of 33 children with HUS were less than a year old, 90% less than five. A constant laboratory finding was the presence of FDP in the blood and urine, and of the remaining tests the presence of erythrocyte fragments. The absence of thrombocytopenia does not eliminate HUS. Antiplatelet treatment was given to 94% of children, heparin to 94%, thrombolysis to 27% Precise evaluation of treatment would require a controlled prospective multicentre study.
Blood Coagulation Tests↗